Patent application title: Novel genes encoding novel proteolytic enzymes
Inventors:
Luppo Edens (Rotterdam, NL)
Alard Van Albertus Dijk (Vlaardingen, NL)
Phillip Krubasik (Martinsried, DE)
Kaj Albermann (Martinsried, DE)
Alexander Stock (Martinsried, DE)
Erik Kimpel (Munchen, DE)
Sabrine Klaubauer (Munchen, DE)
Christian Wagner (Martinsried, DE)
Andreas Fritz (Martinsried, DE)
Wilk Von Gustedt (Planegg, DE)
Oliver Henrich (Martinsried, DE)
Dieter Maier (Martinsried, DE)
Fabio Spreafico (Martinsried, DE)
Ulrike Folkers (Munchen, DE)
Sylvia Hopper (Munchen, DE)
Wolfram Kemmner (Munchen, DE)
Pamela Tan (Martinsfried, DE)
Josephine Stiebler (Munchen, DE)
Ricahrd Albang (Martinsried, DE)
Assignees:
DSM IP ASSETS B.V.
IPC8 Class: AC12N948FI
USPC Class:
435 691
Class name: Chemistry: molecular biology and microbiology micro-organism, tissue cell culture or enzyme using process to synthesize a desired chemical compound or composition recombinant dna technique included in method of making a protein or polypeptide
Publication date: 2009-11-05
Patent application number: 20090275079
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Patent application title: Novel genes encoding novel proteolytic enzymes
Inventors:
Luppo Edens
Andreas Fritz
Dieter Maier
Christian Wagner
Ulrike Folkers
Fabio Spreafico
Alard Van Albertus Dijk
Phillip Krubasik
Kaj Albermann
Alexander Stock
Erik Kimpel
Sabrine Klaubauer
Wilk Von Gustedt
Oliver Henrich
Sylvia Hopper
Wolfram Kemmner
Pamela Tan
Josephine Stiebler
Ricahrd Albang
Agents:
NIXON & VANDERHYE, PC
Assignees:
DSM IP ASSETS B.V.
Origin: ARLINGTON, VA US
IPC8 Class: AC12N948FI
USPC Class:
435 691
Patent application number: 20090275079
Abstract:
The invention relates to newly identified gene sequences that encode novel
proteases obtainable from Aspergillus niger. The invention features the
full length gene sequence of the novel genes, their cDNA sequences as
well as the full-length functional protein and fragments thereof. The
invention also relates to methods of using these enzymes in industrial
processes and methods of diagnosing fungal infections. Also included in
the invention are cells transformed with DNA according to the invention
and cells wherein a protease according to the invention is genetically
modified to enhance or reduce its activity and/or level of expression.Claims:
1. An isolated polynucleotide that encodes a tripeptidyl peptidase wherein
said polynucleotide hybridizes to the full length complement of a
nucleotide sequence selected from the group consisting of SEQ ID NO:1 to
SEQ ID NO:8; SEQ ID NO:11; SEQ ID NO:13 to SEQ ID NO:25; SEQ ID NO:27 to
SEQ ID NO:34; SEQ ID NO:36 to SEQ ID NO:49; SEQ ID NO:51 to SEQ ID NO:57;
SEQ ID NO:58 to SEQ ID NO:65; SEQ ID NO:68; SEQ ID NO:70 to SEQ ID NO:82;
SEQ ID NO:84 to SEQ ID NO:91; SEQ ID NO:93 to SEQ ID NO:106; and SEQ ID
NO:108 to SEQ ID NO:115 under hybridization conditions of 5.times.SSC,
5.times.Denhardt's solution, and 1.0% SDS at 68.degree. C. and wash
conditions of 0.2.times.SSC and 0.1% SDS at room temperature.
2. The isolated polynucleotide of claim 1, which comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:8; SEQ ID NO:11; SEQ ID NO:13 to SEQ ID NO:25; SEQ ID NO:27 to SEQ ID NO:34; SEQ ID NO:36 to SEQ ID NO:49; SEQ ID NO:51 to SEQ ID NO:57; SEQ ID NO:58 to SEQ ID NO:65; SEQ ID NO:68; SEQ ID NO:70 to SEQ ID NO:82; SEQ ID NO:84 to SEQ ID NO:91; SEQ ID NO:93 to SEQ ID NO:106; and SEQ ID NO:108 to SEQ ID NO:115.
3. The isolated polynucleotide of claim 1 obtainable from a filamentous fungus.
4. The isolated polynucleotide of claim 3 obtainable from Aspergillus niger.
5. An isolated polynucleotide encoding a polypeptide having tripeptidyl peptidase-activity, which is at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:115 to SEQ ID NO:122; SEQ ID NO:125; SEQ ID NO:127 to SEQ ID NO:139; SEQ ID NO:141 to SEQ ID NO:148; SEQ ID NO:150 to SEQ ID NO:163; and SEQ ID NO:165 to SEQ ID NO:171.
6. The isolated polynucleotide of claim 7 wherein said polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:115 to SEQ ID NO:122; SEQ ID NO:125; SEQ ID NO:127 to SEQ ID NO:139; SEQ ID NO:141 to SEQ ID NO:148; SEQ ID NO:150 to SEQ ID NO:163; and SEQ ID NO:165 to SEQ ID NO:171.
7. The isolated polynucleotide of claim 1, which hybridizes to the full length of a nucleotide sequence selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:8; SEQ ID NO:11; SEQ ID NO:13 to SEQ ID NO:25; SEQ ID NO:27 to SEQ ID NO:34; SEQ ID NO:36 to SEQ ID NO:49; SEQ ID NO:51 to SEQ ID NO:57; SEQ ID NO:58 to SEQ ID NO:65; SEQ ID NO:68; SEQ ID NO:70 to SEQ ID NO:82; SEQ ID NO:84 to SEQ ID NO:91; SEQ ID NO:93 to SEQ ID NO:106; and SEQ ID NO:108 to SEQ ID NO:115.
8. A vector comprising the polynucleotide sequence of claim 1.
9. The vector of claim 8 wherein said polynucleotide sequence is operatively linked with regulatory sequences suitable for expression of said polynucleotide sequence in a suitable host cell.
10. The vector of claim 9 wherein said suitable host cell is a filamentous fungus.
11. An isolated recombinant host cell comprising the vector of claim 9.
12. A method to prepare a tripeptidyl peptidase comprising the steps of culturing a host cell comprising the vector of claim 9 and isolating said tripeptidyl peptidase from said host cell.
13. A vector comprising the polynucleotide sequence of claim 5.
14. The vector of claim 13 wherein said polynucleotide sequence is operatively linked with regulatory sequences suitable for expression of said polynucleotide sequence in a suitable host cell.
15. The vector of claim 14 wherein said suitable host cell is a filamentous fungus.
16. An isolated recombinant host cell comprising the vector of claim 14.
17. A method to prepare a tripeptidyl peptidase comprising the steps of culturing a host cell comprising the vector of claim 14 and isolating said tripeptidyl peptidase from said host cell.
18. An isolated polypeptide having tripeptidyl peptidase activity, which comprises an amino acid sequence at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:115 to SEQ ID NO:122; SEQ ID NO:125; SEQ ID NO:127 to SEQ ID NO:139; SEQ ID NO:141 to SEQ ID NO:148; SEQ ID NO:150 to SEQ ID NO:163; and SEQ ID NO:165 to SEQ ID NO:171.
19. The isolated polypeptide of claim 18 wherein said polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:115 to SEQ ID NO:122; SEQ ID NO:125; SEQ ID NO:127 to SEQ ID NO:139; SEQ ID NO:141 to SEQ ID NO:148; SEQ ID NO:150 to SEQ ID NO:163; and SEQ ID NO:165 to SEQ ID NO:171.
20. An isolated polypeptide obtainable by expressing the polynucleotide of claim 1 in a host cell.
21. An isolated polypeptide obtainable by expressing the polynucleotide of claim 5 in a host cell.
22. An isolated recombinant host cell expressing the polypeptide of claim 18.
23. A fusion protein comprising the amino acid sequence according to claim 18.
24. A fusion protein comprising the amino acid sequence of the polypeptide according to claim 20.
25. A fusion protein comprising the amino acid sequence of the polypeptide according to claim 21.
Description:
FIELD OF THE INVENTION
[0001]The invention relates to newly identified polynucleotide sequences comprising genes that encode novel proteases isolated from Aspergillus niger. The invention features the full length nucleotide sequence of the novel genes, the cDNA sequences comprising the full length coding sequences of the novel proteases as well as the amino acid sequences of the full-length functional proteins and fragments and variants thereof. The invention also relates to methods of using these enzymes in industrial processes and methods of diagnosing fungal infections. Also included in the invention are cells transformed with a polynucleotide according to the invention and cells wherein a protease according to the invention is genetically modified to enhance or reduce its activity and/or level of expression.
BACKGROUND OF THE INVENTION
Proteolytic Enzymes
[0002]Proteins can be regarded hetero-polymers that consist of amino acid building blocks connected by a peptide bond. The repetitive unit in proteins is the central alpha carbon atom with an amino group and a carboxyl group. Except for glycine, a so-called amino acid side chain substitutes one of the two remaining alpha carbon hydrogen atoms. The amino acid side chain renders the central alpha carbon asymmetric. In general, in proteins the L-enantiomer of the amino acid is found. The following terms describe the various types of polymerized amino acids. Peptides are short chains of amino acid residues with defined sequence. Although there is not really a maximum to the number of residues, the term usually indicates a chain which properties are mainly determined by its amino acid composition and which does not have a fixed three-dimensional conformation. The term polypeptide is usually used for the longer chains, usually of defined sequence and length and in principle of the appropriate length to fold into a three-dimensional structure. Protein is reserved for polypeptides that occur naturally and exhibit a defined three-dimensional structure. In case the proteins main function is to catalyze a chemical reaction it usually is called an enzyme. Proteases are the enzymes that catalyze the hydrolysis of the peptide bond in (poly)peptides and proteins.
[0003]Under physiological conditions proteases catalyse the hydrolysis of the peptide bond. The International Union of Biochemistry and Molecular Biology (1984) has recommended to use the term peptidase for the subset of peptide bond hydrolases (Subclass E.C 3.4.). The terms protease and peptide hydrolase are synonymous with peptidase and may also be used here. Proteases comprise two classes of enzymes: the endo-peptidases and the exo-peptidases, which cleave peptide bonds at points within the protein and remove amino acids sequentially from either N or C-terminus respectively. Proteinase is used as a synonym for endo-peptidase. The peptide bond may occur in the context of di-, tri-, tetra-peptides, peptides, polypeptides or proteins. In general the amino acid composition of natural peptides and polypeptides comprises 20 different amino acids, which exhibit the L-configuration (except for glycine which does not have a chiral centre). However the proteolytic activity of proteases is not limited to peptides that contain only the 20 natural amino acids. Peptide bonds between so-called non-natural amino acids can be cleaved too, as well as peptide bonds between modified amino acids or amino acid analogues. Some proteases do accept D enantiomers of amino acids at certain positions. In general the remarkable stereo-selectivity of proteases makes them very useful in the process of chemical resolution. Many proteases exhibit interesting side activities such as esterase activity, thiol esterase activity and (de)amidase activity. These side activities are usually not limited to amino acids only and might turn out to be very useful in bioconversions in the area of fine chemicals.
[0004]There are a number of reasons why proteases of filamentous fungi, eukaryotic microorganisms, are of particular interest. The basic process of hydrolytic cleavage of peptide bonds in proteins appears costly and potentially detrimental to an organism if not properly controlled. The desired limits to proteolytic action are achieved through the specificity of proteinases, by compartmentalization of proteases and substrates within the cell, through modification of the substrates allowing recognition by the respective proteases, by regulation via zymogen activation, and the presence or absence of specific inhibitors, as well through the regulation of protease gene expression. In fungi, proteases are also involved in other fundamental cellular processes, including intracellular protein turnover, processing, translocation, sporulation, germination and differentiation. In fact, Aspergillus nidulans and Neurospora crassa have been used as model organisms for analyzing the molecular basis of a range of physiological and developmental processes. Their genetics enable direct access to biochemical and genetical studies, under defined nutrient and cultivation conditions. Furthermore, a large group of fungi pathogenic to humans, live-stock and crop, has been isolated and proteolysis has been suggested to play a role in their pathogenicity (host penetration, countering host defense mechanisms and/or nutrition during infection). Proteases are also frequently used in laboratory, clinical and industrial processes; both microbial and non-microbial proteases are widely used in the food industry (baking, brewing, cheese manufacturing, meat tenderizing), in tanning industry and in the manufacture of biological detergents (Aunstrup, 1980). The commercial interest in exploiting certain filamentous fungi, especially the Aspergilli, as hosts for the production of both homologous and heterologous proteins, has also recently renewed interests in fungal proteases (van Brunt, 1986ab). Proteases often cause problems in heterologous expression and homologous overexpression of proteins in fungi. In particular, heterologous expression is hampered by the proteolytic degradation of the expressed products by homologous proteases. These commercial interests have resulted in detailed studies of proteolytic spectra and construction of protease deficient strains and have improved the knowledge about protease expression and regulation in these organisms. Consequently there is a great need to identify and eliminate novel proteases in filamentous fungi.
[0005]Micro-organisms such as for example fungi are particularly useful in the large scale production of proteins. In particular when such proteins are secreted into the medium. Proteolytic enzymes play a role in these production processes. On the one hand particular proteolytic enzymes are in general required for proper processing of the target protein and the metabolic well-being of the production host. On the other hand proteolytic degradation may significantly decrease the yield of secreted proteins. Poor folding in the secretion pathway may lead to degradation by intracellular proteases. This might be a particular problem with producing heterologous proteins. The details of the proteolytic processes, which are responsible for the degradation of the proteins that are diverted from the secretory process in fungi are not exactly known. In eukaryotes the degradation of cellular proteins is achieved by a proteasome and usually involves ubiquitin labelling of proteins to be degraded. In fungi, proteasomal and vacuolar proteases are also likely candidates for the proteolytic degradation of poorly folded secretory proteins. The proteolytic degradation is likely cytoplasmic, but endoplamatic reticulum resident proteases cannot be excluded. From the aspect of production host strain improvement the proteolytic system may be an interesting target for genetic engineering and production strain improvement. Additional copies of protease genes, over-expression of certain proteases, modification of transcriptional control, as well as knock out procedures for deletion of protease genes may provide a more detailed insight in the function a given protease. Deletion of protease encoding genes can be a valuable strategy for host strain improvement in order to improve production yield for homologous as well as heterologous proteins.
[0006]Eukaryotic microbial proteases have been reviewed by North (1982). More recently, Suarez Rendueles and Wolf (1988) have reviewed the S. cerevisiae proteases and their function.
[0007]Apart from the hydrolytic cleavage of bonds, proteases may also be applied in the formation of bonds. Bonds in this aspect comprise not only peptide and amide bonds but also ester bonds. Whether a protease catalyses the cleavage or the formation of a particular bond does in the first place depend on the thermodynamics of the reaction. An enzyme such as a protease does not affect the equilibrium of the reaction. The equilibrium is dependent on the particular conditions under which the reaction occurs. Under physiological conditions the thermodynamics of the reactions is in favour of the hydrolysis of the peptide due to the thermodynamically very stable structure of the zwitterionic product. By application of physical-chemical principles to influence the equilibrium, or by manipulating the concentrations or the nature of the reactants and products, or by exploiting the kinetic parameters of the enzyme reaction it is possible to apply proteases for the purpose of synthesis of peptide bonds. The addition of water miscible organic solvents decreases the ektent of ionisation of the carboxyl component, thereby increasing the concentration of substrate available for the reaction. Biphasic systems, water mimetics, reverse micelles, anhydrous media, or modified amino and carboxyl groups to invoke precipitation of products are often employed to improve yields. When the proteases with the right properties are available the application of proteases for synthesis offers substantial advantages. As proteases are stereo-selective as well as regio-selective, sensitive groups on the reactants do usually not need protection and reactants do not need to be optically pure. As conditions of enzymatic synthesis are mild, racemization and decomposition of labile reactants or products can be prevented. Apart from bonds between amino acids, also other compounds exhibiting a primary amino group, a thiol group or a carboxyl group may be linked by properly selected proteases. In addition esters, thiol esters and amides may be synthesized by certain proteases. Protease have been shown to exhibit regioselectively in the acylation of mono, di- and tri-saccharides, nucleosides, and riboflavin. Problems with stability under the sometimes harsh reaction conditions may be prevented by proper formulation. Encapsulation and immobilisation do not only stabilise enzymes but also allow easy recovery and separation from the reaction medium. Extensive crosslinking, treatment with aldehydes or covering the surface with certain polymers such as dextrans, polyethyleneglycol, polyimines may substantially extend the lifetime of the biocatalyst.
The Natural Roles of Proteases
[0008]Traditionally, proteases have been regarded as degrading enzymes, capable of cleaving proteins into small peptides and/or amino acids, and whose role it is to digest nutrient protein or to participate in the turnover of cellular proteins. In addition, it has been shown that proteases also play key roles in a wide range of cellular processes, via mechanisms of selective modification by limited proteolysis, and thus can have essential regulatory functions (Holzer and Tschensche 1979; Holzer and Heinrich, 1980). The specificity of a proteinase is assumed to be closely related to its physiological function and its mode of expression. With respect to the function of a particular protease, its localisation is often very important; for example, a lot of the vacuolar and periplasmic proteases are involved in protein degradation, while many of the membrane-bound proteases are important in protein processing (Suarez Rendueles and Wolf, 1988). The different roles of proteases in many cellular processes can be divided into four main functions of proteases: 1) protein degradation, 2) posttranslational processing and (in)activation of specific proteins, 3) morphogenesis, and 4) pathogenesis.
[0009]An obvious role for proteases in organisms which utilise protein as a nutrient source is in the hydrolysis of nutrients. In fungi, this would involve the degradation outside the cells by extracellular broad specificity proteases. Protein degradation is also important for rapid turnover of cellular proteins and allows the cell to remove abnormal proteins and to adapt their complement of protein to changing physiological conditions. Generally, proteases of rather broad specificity should be extremely well-controlled in order to protect the cell from random degradation of other than correct target proteins.
[0010]Contrary to the hydrolysis the synthesis of polypeptides occurs in vivo by an ATP driven process on the ribosome. Ultimately the sequence in which the amino acids are linked is dictated by the information derived from the genome. This process is known as the transcription. Primary translation products are often longer than the final functional products, and after the transcription usually further processing of such precursor proteins by proteases is required. Proteases play a key role in the maturation of such precursor proteins to obtain the final functional protein. In contrast to the very controlled trimming and reshaping of proteins, proteases can also be very destructive and may completely degrade polypeptides into peptides and amino acids. In order to avoid that proteolytic activity is unleashed before it is required, proteases are subject to extensive regulation. Many proteases are synthesized as larger precursors known as zymogens, which become activated when required. Remarkably this activation always occurs by proteolysis. Apart from direct involvement in the processing, selective activation and inactivation of individual proteins are well-known phenomena catalyzed by specific proteases.
[0011]The selectivety of limited proteolysis appears to reside more directly in the proteinase-substrate interaction. Specificity may be derived from the proteolytic enzyme which recognizes only specific amino acid target sequences. On the other hand, it may also be the result of selective exposure of the `processing site` under certain conditions such as pH, ionic strength or secondary modifications, thus allowing an otherwise non-specific protease to catalyze a highly specific event. The activation of vacuolar zymogens by limited proteolysis gives an example of the latter kind.
[0012]Morphogenesis or differentiation can be defined as a regulated series of events leading to changes from one state to another in an organism. Although direct relationships between proteases and morphological effects could not be established in many cases, the present evidence suggests a significant involvement of proteases in fungal morphogenesis; apart form the observed extensive protein turnover during differentiation, sporulation and spore germination, proteases are thought to be directly involved in normal processes as hyphal tip branching and septum formation, (Deshpande, 1992).
[0013]Species of Aspergillus, in particular A. fumigatus and A. flavus, have been implicated as the causative agents of a number of diseases in humans and animals called aspergillosis (Bodey and Vartivarian, 1989). It has been repeatedly suggested that proteases are involved in virulence of A. fumigatus and A. flavus like there are many studies linking secreted proteases and virulence of bacteria. In fact, most human infections due to Aspergillus species are characterised by an extensive degradation of the parenchyma of the lung which is mainly composed of collagen and elastin (Campbell et al., 1994). Research has been focussed on the putative role of the secreted proteases in virulence of A. fumigatus and A. flavus which are the main human pathogens and are known to possess elastinolytic and collagenic activities (Kolattukudy et al., 1993). These elastinolytic activities were shown to correlate in vitro with infectivity in mice (Kothary et al., 1984). Two secreted proteases are known to be produced by A. fumigatus and A. flavus, an alkaline serine protease (ALP) and a neutral metallo protease (MEP). In A. fumigatus both the genes encoding these proteases were isolated, characterised and disrupted (Reicherd et al., 1990; Tang et al, 1992, 1993; Jaton-Ogay et al., 1994). However, alp mep double mutants showed no differences in pathogenecity when compared with wild type strains. Therefore, it must be concluded that the secreted A. fumigatus proteases identified in vitro are not essential factors for the invasion of tissue (Jaton Ogay et al., 1994). Although A. fumigatus accounts for only a small proportion of the airborne mould spores, it is the most frequently isolated fungus from lung and sputem (Schmitt et al., 1991). Other explanations for the virulence of the fungus could be that the conditions in the bronchia (temperature and nutrients) are favourable for the parasitic growth of A. fumigatus. As a consequence, invasive apergillosis could be a circumstancial event, when the host pathogenic defences have been weakened by immunosuppressive treatments or diseases like AIDS.
[0014]Four major classes of proteases are known and are designated by the principal functional groups in their active site: the `serine`, the `thiol` or `cysteine`, the `aspartic` or `carboxyl` and the `metallo` proteases. A detailed state of the art review on these major classes of proteases, minor classes and unclassified proteases can be found in Methods in Enzymology part 244 and 248 (A. J. Barrett ed, 1994 and 1995).
Specificity of Proteases
[0015]Apart from the catalytic machinery of proteases another important aspect of proteolytic enzymes is the specificity of proteases. The specificity of a protease indicates which substrates the protease is likely to hydrolyze. The twenty natural amino acids offer a large number of possibilities to make up peptides. Eg with twenty amino acids one can make up already 400 dipeptides and 800 different tripeptide, and so on. With longer peptides the number of possibilities will become almost unlimited. Certain proteases hydrolyze only particular sequences at a very specific position. The interaction of the protease with the peptide substrate may encompass one up to ten amino acid residues of the peptide substrate. With large proteinacious substrates there may be even more residues of the substrate that interact with the proteases. However this likely involves less specific interactions with protease residues outside the active site binding cleft. In general the specific recognition is restricted to the linear peptide, which is bound in the active site of the protease.
[0016]The nomenclature to describe the interaction of a substrate with a protease has been introduced in 1967 by Schechter and Berger (Biochem. Biophys. Res. Com., 1967, 27, 157-162) and is now widely used in the literature. In this system, it is considered that the amino acid residues of the polypeptide substrate bind to so-called sub-sites in the active site. By convention, these sub-sites on the protease are called S (for sub-sites) and the corresponding amino acid residues are called P (for peptide). The amino acid residues of the N-terminal side of the scissile bond are numbered P3, P2, P1 and those residues of the C-terminal side are numbered P1', P2', P3'. The P1 or P1' residues are the amino acid residues located near the scissile bond. The substrate residues around the cleavage site can then be numbered up to P8. The corresponding sub-sites on the protease that complement the substrate binding residues are numbered S3, S2, S1, S1', S2', S3', etc, etc. The preferences of the sub-sites in the peptide binding site determine the preference of the protease for cleaving certain specific amino acid sequences at a particular spot. The amino acid sequence of the substrate should conform with the preferences exhibited by the sub-sites. The specificity towards a certain substrate is clearly dependant both on the binding affinity for the substrate and on the velocity at which subsequently the scissile bond is hydrolysed. Therefore the specificity of a protease for a certain substrate is usually indicated by its kcat/Km ratio, better known as the specificity constant. In this specificity constant kcat represents the turn-over rate and Km is the dissociation constant.
[0017]Apart from amino acid residues involved in catalysis and binding, proteases contain many other essential amino acid residues. Some residues are critical in folding, some residues maintain the overall three dimensional architecture of the protease, some residues may be involved in regulation of the proteolytic activity and some residue may target the protease for a particular location. Many proteases contain outside the active site one or more binding sites for metal ions. These metal ions often play a role in stabilizing the structure. In addition secreted eukaryotic microbial proteases may be extensively glycosylated. Both N- and O-linked glycosylation occurs. Glycosylation may aid protein folding, may increase solubility, prevent aggregation and as such stabilize the mature protein. In addition the extent of glycosylation may influence secretion as well as water binding by the protein.
Regulation of Proteolytic Activity
[0018]A substantial number of proteases are subject to extensive regulation of the proteolytic activity in order to avoid undesired proteolytic damage. To a certain extent this regulation takes place at transcription level. For example in fungi the transcription of secreted protease genes appears to be sensitive to external carbon and nitrogen sources, whereas genes encoding intracellular proteases are insensitive. The extracellular pH is sensed by fungi and some genes are regulated by pH. In this process transcriptional regulator proteins play a crucial role. Proteolytic processing of such regulator proteins is often the switch that turns the regulator proteins either on or off.
[0019]Proteases are subject to intra- as well as intermolecular regulation. This implies certain amino acids in the proteolytic enzyme molecule that are essential for such regulation. Proteases are typically synthesized as larger precursors known as zymogens, which are catalytically inactive. Usually the peptide chain extension rendering the precursor protease inactive is located at the amino terminus of the protease. The precursor is better known as pro-protein. As many of the proteases processed in this way are secreted from the cells they contain in addition a signal sequence (pre sequence) so that the complete precursor is synthesized as a pre-pro-protein. Apart from rendering the protease inactive the pro-peptide often is essential for mediating productive folding. Examples of proteases include serine proteases (alpha lytic protease, subtilisin, aqualysin, prohormone convertase), thiol proteases (cathepsin L and cruzian), aspartic proteases (proteinase A and cathepsin D) and metalloproteases. In addition the pro-peptide might play a role in cellular transport either alone or in conjunction with signal peptides. It may facilitate interaction with cellular chaperones or it may facilitate transport over the membrane. The size of the extension in the precursor pre-pro-protein may vary substantially, ranging from a short peptide fragment to a polypeptide, which can exist as an autonomous folding unit. In particular these larger extensions are often observed to be strong inhibitors of the protease even after cleavage from the protease. It was observed that even after cleavage such pro-peptides could assist in proper folding of the proteases. As such pro-peptides can be considered to function as molecular chaperones and separate or additional co-expression of such pro-peptides could be advantageous for protease production.
[0020]There is substantial difference in the level of regulation between proteases that are secreted into the medium and proteases that remain intracellular. Proteases secreted into the medium are usually after activation no longer subject to control and therefore are usually relatively simple in their molecular architecture consisting of one globular module. Intracellular proteases are necessarily subject to continuous control in order to avoid damage to the cells. In contrast with zymogens of secreted proteases in more complex regulatory proteases very large polypeptide segments may be inserted between the signal and the zymogen activation domain of the proteolytic module. Structure-function studies indicate that such non-protease parts may be involved in interactions with macroscopic structures, membranes, cofactors, substrates, effectors, inhibitors, ions, that regulate activity and activation of the proteolytic module(s) or its (their) zymogens. The non-proteolytic modules exhibit remarkable variation in size and structure. Many of the modules can exist as such independently from the proteolytic module. Therefore such modules can be considered to correspond to independent structural and functional units that are autonomous with respect to folding. The value of such a modular organization is that acquisition of new modules can endow the recipient protease with new novel binding specificities and can lead to dramatic changes in its activity, regulation and targeting. The principle of modular organized proteolytic enzymes may also be exploited by applying molecular biology tools in order to create novel interactions, regulation, specificity, and/or targeting by shuffling of modules. Although in general such additional modules are observed as N or C terminal extension, also large insertions within the exterior loops of the catalytic domain have been observed. It is believed that also in this case the principal fold of the protease represents still the essential topology to form a functional proteolytic entity and that the insertion can be regarded as substructure folded onto the surface of the proteolytic module.
Molecular Structure
[0021]In principle the modular organization of larger proteins is a general theme in nature. In particular within the larger multimodular frameworks typical proteolytic modules show sizes of 100 to 400 amino acids on the average. This corresponds with the average size of most of the globular proteolytic enzymes that are secreted into the medium. As discussed above polypeptide modules are polypeptide fragments, which can fold and function as independent entities. Another term for such modules is domains. However domain is used in a broader context than module. The term domain as used herein refers usually to a part of the polypeptide chain that depicts in the three-dimensional structure a typical folding topology. In a protein domains interact to varying extents, but less extensively than do the structural elements within domains. Other terms such as subdomain and folding unit are also used in literature. As such it is observed that many proteins that share a particular functionality may share the same domains. Such domains can be recognized from the primary structure that may show certain sequence patterns, which are typical for a particular domain. Typical examples are the mononucleotide binding fold, cellulose binding domains, helix-turn-helix DNA binding motif, zinc fingers, EF hands, membrane anchors. Modules refer to those domains which are expected to be able to fold and function autonomously. A person skilled in the art knows how to identify particular domains in a primary structure by applying commonly available computer software to said structure and homologous sequences from other organisms or species.
[0022]Although multimodular or multidomain proteins may appear as a string of beads, assemblies of substantial more complex architecture have been observed. In case the various beads reside on the same polypeptide chain the beads are generally called modules or domains. When the beads do not reside on one and same polypeptide chain but form assemblies via non-covalent interactions then the term subunit is used to designate the bead. Subunits may be transcribed by one and the same gene or by different genes. The multi-modular protein may become proteolytically processed after transcription leading to multiple subunits. Individual subunits may consist of multiple domains. Typically the smaller globular proteins of 100-300 amino acids usually consist only of one domain.
Molecular Classification of Proteolytic Enzymes
[0023]In general proteases are classified according to their molecular properties or according to their functional properties. The molecular classification is based on the primary structure of the protease. The primary structure of a protein represents its amino acid sequence, which can be derived from the nucleotide sequence of the corresponding gene. Tracing extensively the similarities in the primary structures may allow for the notice of similarities in catalytic mechanism and other properties, which even may extend to functional properties. The term family is used to describe a group of proteases that show evolutionary relationship based on similarity between their primary structures. The members of such a family are believed to have arisen by divergent evolution from the same ancestor. Within a family further sub-grouping of the primary structures based on more detailed refinement of sequence comparisons results in subfamilies. Classification according to three-dimensional fold of the proteases may comprise secondary structure, tertiary structure and quarternary structure. In general the classification on secondary structure is limited to content and gross orientation of secondary structure elements. Similarities in tertiary structure have led to the recognition of superfamilies or clans. A superfamily or a clan is a group of families that are thought to have common ancestry, as they show a common 3-dimensional fold. In general tertiary structure is more conserved than the primary structure. As a consequence similarity of the primary structure does not always reflect similar functional properties. In fact functional properties may have diverged substantially resulting in interesting new properties. At present quarternary structure has not been applied to classify various proteases. This might be due to a certain bias of the structural databases towards simple globular proteases. Many proteolytic systems that are subject to activation, regulation, or complex reaction cascades are likely to consist of multiple domains or subunits. General themes in the structural organization of such protease systems may lead to new types of classification.
Classification According to Specificity.
[0024]In absence of sequence information proteases haven been subject to various type of functional classification. The classification and naming of enzymes by reference to the reactions which are catalyzed is a general principle in enzyme nomenclature. This approach is also the underlying principle of the EC numbering of enzymes (Enzyme Nomenclature 1992 Academic Press, Orlando). Two types of proteases (EC 3.4) can be recognized within Enzyme Nomenclature 1992, those of the exo-peptidases (EC 3.4.11-19) and those of the endo-peptidases (EC 3.4.21-24, 3.4.99). Endo-peptidases cleave peptide bonds in the inner regions of the peptide chain, away from the termini. Exo-peptidases cleave only residues from the ends of the peptide chain. The exo-peptidases acting at the free N-terminus may liberate a single amino acid residue, a dipeptide or a tripeptide and are called respectively amino peptidases (EC 3.4.11), dipeptidyl peptidases (EC 3.4.14) and tripeptidyl peptidase (EC 3.3.14). Proteases starting peptide processing from the carboxyl terminus liberating a single amino acid are called carboxy peptidase (EC 3.4.16-18). Peptidyl-dipeptidases (EC 3.4.15) remove a dipeptide from the carboxyl terminus. Exo- and endo-peptidase in one are the dipeptidases (EC 3.4.13), which cleave specifically only dipeptides in their two amino acid halves. Omega peptidases (EC 3.4.19) remove terminal residues that are either substituted, cyclic, or linked by isopeptide bonds
[0025]Apart from the position where the protease cleaves a peptide chain, for each type of protease a further division is possible based on the nature of the preferred amino acid residues in the substrate. In general one can distinguish proteases with broad, medium and narrow specificity. Some proteases are simply named after the specific proteins or polypeptides that they hydrolyze, e.g. keratinase, collagenase, elastase. A narrow specificity may pin down to one particular amino acid or one particular sequence which is removed or which is cleaved respectively. When the protease shows a particular preference for one amino acid in the P1 or P1' position the name of this amino acid may be a qualifier. For example prolyl amino peptidase removes proline from the amino terminus of a peptide (proline is the P1 residue). X-Pro or proline is used when the bond on the imino side of the proline is cleaved (proline is P1' residue), eg proline carboxypeptidase removes proline from the carboxyl terminus. Prolyl endopeptidase (or Pro-X) cleaves behind proline while proline endopeptidase (X-Pro) cleaves in front of a proline. Amino acid residue in front of the scissile peptide bond refers to the amino acid residue that contributes the carboxyl group to the peptide bond. The amino acids residue behind the scissile peptide bond refers to the amino acid residue that contributes the amino group to the peptide bond. According to the general convention an amino acid chain runs from amino terminus (the start) to the carboxyl terminus (the end) and is numbered accordingly. Endo proteases may also show clear preference for a particular amino acid in the P1 or P1' position, eg glycyl endopeptidase, peptidyl-lysine endopeptidase, glutamyl endopeptidase. In addition proteases may show a preference for a certain group of amino acids that share a certain resemblance. Such a group of preferred amino acids may comprise the hydrophobic amino acids, only the bulky hydrophobic amino acids, small hydrophobic, or just small amino acids, large positively charged amino acids, etc, etc. Apart from preferences for P1 and P1' residues also particular preferences or exclusions may exist for residues preferred by other subsites on the protease. Such multiple preferences can result in proteases that are very specific for only those sequences that satisfy multiple binding requirements at the same time. In general it should be realized that protease are rather promiscuous enzymes. Even very specific protease may cleave peptides that do not comply with the generally observed preference of the protease. In addition it should be realized that environmental conditions such as pH, temperature, ionic strength, water activity, presence of solvents, presence of competing substrates or inhibitors may influence the preferences of the proteases. Environmental condition may not only influence the protease but also influence the way the proteinacious substrate is presented to the protease.
Classification by Catalytic Mechanism.
[0026]Proteases can be subdivided on the basis of their catalytic mechanism. It should be understood that for each catalytic mechanism the above classification based on specificity leads to further subdivision for each type of mechanism. Four major classes of proteases are known and are designated by the principal functional group in the active site: the serine proteases (EC 3.4.21 endo peptidase, EC 3.4.16 carboxy peptidase), the thiol or cysteine proteases (EC 3.4.22 endo peptidase, EC 3.4.18 carboxy peptidase), the carboxyl or aspartic proteases (EC 3.4.23 endo peptidase) and metallo proteases (EC 3.4.24 endo peptidase, EC 3.4.18 carboxy peptidase). There are characteristic inhibitors of the members of each catalytic type of protease. These small inhibitors irreversibly modify an amino acid residue of the protease active site. For example, the serine protease are inactivated by Phenyl Methane Sulfonyl Fluoride (PMSF) and Diisopropyl Fluoro Phosphate (DFP), which react with the active Serine whereas the chloromethylketone derivatives react with the Histidine of the catalytic triad. Phosphoramidon and 1,10 Phenanthroline typically inhibit metallo proteases. Inhibition by Pepstatin generally indicates an aspartic protease. E64 inhibits thiol protease specifically. Amastatin and Bestatin inhibit various aminopeptidases. Substantial variations in susceptibility of the proteases to the inhibitors are observed, even within one catalytic class. To a certain extent this might be related to the specificity of the protease. In case binding site architecture prevents a mechanism based inhibitor to approach the catalytic site, then such a protease escapes from inhibition and identification of the type of mechanism based on inhibition is prohibited. Chymostation for example is a potent inhibitor for serine protease with chymotrypsin like specificity, Elastatinal inhibits elastase like serine proteases and does not react with trypsin or chymostrypsin, 4 amido PMSF (APMSF) inhibits only serine proteases with trypsin like specificity. Extensive accounts of the use of inhibitors in the classification of proteases include Barret and Salvesen, Proteinase Inhibitors, Elsevier Amstardam, 1986; Bond and Beynon (eds), Proteolytic Enzymes, A Practical Approach, IRL Press, Oxford, 1989; Methods in Enzymology, eds E. J. Barret, volume 244, 1994 and volume 248, 1995; E. Shaw, Cysteinyl proteinases and their selective inactivation, Adv Enzymol. 63:271-347 (1990)
Classification According to Optimal Performance Conditions.
[0027]The catalytic mechanism of a proteases and the requirement for its conformational integrity determine mainly the conditions under which the protease can be utilized. Finding the protease that performs optimal under application conditions is a major challenge. Often conditions at which proteases have to perform are not optimal and do represent a compromise between the ideal conditions for a particular application and the conditions which would suit the protease best. Apart from the particular properties of the protease it should be realized that also the presentation of a proteinacious substrates is dependant on the conditions, and as such determines also which conditions are most effective for proteolysis. Specifications for the enzyme that are relevant for application comprise for example the pH dependence, the temperature dependence, sensitivity for or the dependence of metal ions, ionic strength, salt concentration, solvent compatibility. Another factor of major importance is the specific activity of a protease. The higher the enzyme's specific activity, the less enzyme is needed for a specific conversion. Lower enzyme requirements imply lower costs and lower protein contamination levels.
[0028]The pH is a major parameter that determines protease performance in an application. Therefor pH dependence is an important parameter to group proteases. The major groups that are recognized are the acid proteases, the neutral proteases, the alkaline proteases and the high alkaline proteases. The optimum pH matches only to some extent the proteolytic mechanism, eg aspartic protease show often an optimum at acidic pH, metalloproteases and thiol proteases often perform optimal around neutral pH to slightly alkaline, serine peptidases are mainly active in the alkaline and high alkaline region. For each class exceptions are known. In addition the overall water activity of the system plays a role. The pH optimum of a protease is defined as the pH range where the protease exhibits an optimal hydrolysis rate for the majority of its substrates in a particular environment under particular conditions. This range can be narrow, e.g. one pH unit, as well as quite broad, 3-4 pH units. In general the pH optimum is also dependant on the nature of the proteinacious substrate. Both the turnover rate as well as the specificity may vary as a function of pH. For a certain efficacy it can be desirable to use the protease far from its pH optimum because production of less desired peptides is avoided. Less desired peptides might be for example very short peptides or peptides causing a bitter taste. In addition a more narrow specificity can be a reason to choose conditions that deviate from optimal conditions with respect to turnover rate. Dependant on the pH the specificity may be narrow, e.g. only cleaving the peptide chain in one particular position or before or after one particular amino acid, or broader, e.g. cleaving a chain at multiple positions or cleaving before or after more different types of amino acids. In fact the pH dependence might be an important tool to regulate the proteolytic activity in an application. In case the pH shifts during the process the proteolysis might cease spontaneously without the need for further treatment to inactivate the protease. In some cases the proteolysis itself may be the driver of the pH shift.
[0029]Very crucial for application of proteases is their handling and operating stability. As protease stability is strongly affected by the working temperature, stability is often also referred to as thermostability. In general the stability of a protease indicates how long a protease retains its proteolytic activity under particular conditions. Particular conditions may comprise fermentation conditions, conditions during isolation and down stream processing of the enzyme, storage conditions, formulation and operating or application conditions. In case particular conditions encompass elevated temperatures stability in general refers to thermostability. Apart from the general causes for enzyme inactivation such as chemical modification, unfolding, aggregation etc, main problem with proteases is that they are easy subject to autodegradation. Especially for the utilization of proteases the temperature optimum is a relevant criterion to group proteases. Although there are different definitions, economically the most useful definition is the temperature or the temperature range in which the protease is most productive in a certain application. Protease productivity is a function of both the stability and the turnover rate. Where elevated temperature in general will increase the turnover rate, rapid inactivation will counteract the increase in turnover rate and ultimately lead to low productivity. The conformational stability of the protease under a given process condition will determine its maximum operating temperature. The temperature at which the protease looses it active conformation, often indicated as unfolding or melting point, can be determined according various methods, for example NMR, Circular Dichroism Spectroscopy, Differential Scanning Calorimetry etc. For protease unfolding is usually accompanied by a tremendous increase in autodegradation rate.
[0030]In applications where low temperatures are required protease may be selected with emphasis on a high intrinsic activity at low to moderate temperature. As under such conditions inactivation is relatively slow, under these conditions activity might largely determine productivity. In processes where only during a short period protease activity is required, the stability of the protease might be used as a switch to turn the protease off. In such case more labile instead of very thermostable protease might be preferred.
[0031]Other environmental parameters which may play a role in selecting the appropriate protease may be its sensitivity to salts. The compatibility with metal ions which are found frequently at low concentrations in various natural materials can be crucial for certain applications. In particular with metallo proteases certain ions may replace the catalytic metal ion and reduce or even abolish activity completely. In some applications metal ions have to be added on purpose in order to prevent the washout of the metal ions coordinated to the protease. It is well known that for the sake of enzyme stability and life-time, calcium ions have to be supplied in order to prevent dissociation of protein bound calcium.
[0032]Most microorganisms show a certain tolerance with respect to adapting to changes in the environmental condition. As a consequence at least the proteolytic spectrum that the organism is able to produce are likely to show at least similar tolerances. Such a proteolytic spectrum might be covered by many proteases covering together the hole spectrum or by only a few proteases of a broad spectrum. Taking into account the whole proteolytic spectrum of a microorganism it can be very important to take the location into account.
Cellular Localisation and Characterization of Proteolytic Processing and Degradation
[0033]From an industrial point of view the proteases which are excreted from the cell have specific advantages with respect to producibility at a large scale and stress tolerance as they have to survive without protection of the cell. The large group of cellular protease can be further subdivided in soluble and membrane bound. Membrane bound may comprise protease at the inside as well the outside of the membrane. Intracellular soluble protease may be subdivided further according to specific compartments of the cell where they do occur. As the cell shields the proteases to some extent from the environment and because the cell controls the conditions in the cell, intracellular protease might be more sensitive to large environmental changes and their optima might correlate better with the specific intracelluar conditions. Knowing the conditions of the cellular department where the protease resides might indicate their preferences. Where extracellular protease in general do not require any regulation any more once excreted from the cell, intracellular proteases are often subject to more complicated control and regulation.
[0034]With respect to the function of a particular protease, its localisation is often very important; for example, a lot of the vacuolar and periplasmic proteases are involved in protein degradation, while many of the membrane-bound proteases are important in protein processing (Suarez Rendueles and Wolf, 1988).
[0035]A comprehensive review on the biological properties and evolution of proteases has been published in van den Hombergh: Thesis Landbouwuniversiteit Wageningen: An analysis of the proteolytic system in Aspergillus in order to improve protein production ISBN 90-5485-545-2, which is hereby incorporated by reference herein.
The Protease Problem
[0036]An important reason for the interest in microbial proteases are protease related expression problems observed in several expression hosts used in bioprocess industry. The increasing use of heterologous hosts for the production of proteins, by recombinant DNA technology, has recently brought this problem into focus, since it seems that heterologous proteins are more prone to proteolysis (Archer et al., 1992; van den Hombergh et al., 1996b).
[0037]In S. cerevisiae, already in the early eighties the protease problem and the involvement of several proteases, thus complicating targeted gene disruption approaches to overcome this problem, was recognised. During secretion a protein is exposed to several proteolytic activities residing in the secretory pathway. Additionally, in a prototrophic microorganism as Aspergillus secreted proteins can be exposed to several extracellular proteolytic activities
[0038]The problem of degradation of heterologously expressed proteins is well documented in Aspergillus (van den Hombergh Thesis Landbouwuniversiteit Wageningen: An analysis of the proteolytic system in Aspergillus in order to improve protein production ISBN 90-5485-545-2) and has been reported in the expression of cow prochymosin, human interferon α-2 tPA, GMCSF, IL6, lactoferrin, chicken egg-white lysosyme, porcine plA2, A. niger pectin lyase B, E. coli enterotoxin B and β-glucoronidase, and Erwinia carotovora pectate lyase 3.
[0039]The problem of proteolysis may be addressed at several stages in protein production. Bioprocess engineers may address the problem of proteolysis by downstream processing at low temperatures, by early separation of product and protease(s) or by use of protease inhibitors. These may all lead to successful reduction of the problem. However it is certainly not eliminated, because much of the degradation occurs in vivo during the production of the protein.
[0040]In understanding how proteolysis is controlled in the cell, a major question concerns the recognition mechanism by which proteolysis is triggered. Into what extent are proteolytically susceptable (heterologous) proteins recognised as aberrant because of misfolding or, if correctly folded, as `foreign`, because they do not posses features essential for stability which are specific to the host. Various types of stress can cause the overall proteolysis in a cell to increase significantly. Factors known to increase rate of proteolysis include nutrient starvation and various other types of stress (i.e. elevation of temperature, osmotic stress, toxic substances and expression of certain heterologous proteins). To deal with proteolysis-related expression problems in vivo, several approaches have been proven successful as will be discussed below. However, we have to keep in mind that true `non-proteolytic cells` cannot exist, since proteolysis by intracellular proteases is involved in many essential metabolic and `housekeeping` reactions. Reducing proteolysis will therefore always be a process in which the changed genetical background which results in decreased proteolytic has to be analysed for potential secondary effects which could lead to reduced protein production (e.g. reduced growth rate or sporulation).
Disruption of Proteases in Filamentous Fungal Expression Hosts
[0041]Berka and coworkers (1990) describe the cloning and disruption of the A. awamori pepA gene. More recently, three disrupted aspartyl proteases in A. niger have been described. Disruptants for both the major extracellular aspartyl proteases and the major vacuolar aspartyl protease were described. Double and triple disruptants were generated via recombination and tested for protease spectra and expression and secretion of the A. niger pectin lyase PELB protein, which is very susceptable to proteolytic degradation (van den Hombergh et al., 1995). Disruption of pepA and pepB resulted both in reduction of extracellular protease activities, 80% and 6%, respectively. In the ΔpepE disruptant also other (vacuolar) protease activities were severely affected caused by inactivating of the proteolytic cascade for other vacuolar proteases. Reduced extracellular activities correlated with reduced in vitro degradation of PELB and improved in vivo expression of pelB (van den Hombergh et al., 1996f).
Protease Deficient (prt) Mutants Filamentous Fungi
[0042]Several Aspergillus protease deficient mutants have been studied whether protein production is improved. Archer and coworkers describe the reduced proteolysis of Hen egg white lysozyme in supernatants of an A. niger double prt mutant generated by Mattern and coworkers (1992) and conclude that although the degradation is not absent, it is significantly reduced. Van den Hombergh et al. (1995) show that the in vitro degradation of A. niger PELB is reduced in all seven prt complementation groups they have isolated. Virtually no degradation is observed in the prtB, prtF and prtG mutants. Recently, the expression of the pelB gene was shown to be improved in six complementation groups tested (prtA-F) and highest expression levels were observed in the prtB, prtF and prtG mutants. In addition to the single mutants, which contained residual extracellular proteolytic activities varying from 2-80% compared to wild type activity, double mutants were generated both by recombination and by additional rounds of mutagenesis. Via this approach several double prt mutants were selected and further characterised, which showed a further reduction of PELB degradation compared to their parental strains.
[0043]Instead of elimination of protease activities via disruption or mutagenesis, reduced proteolysis can also be achieved via down-regulation of the interfering proteolytic activities. This may be achieved by genetically altering the promoter or other regulatory sequences of the gene. As shown by Fraissinet-Tachet and coworkers (1996) the extracellular proteases in A. niger are all regulated by carbon catabolite repression and nitrogen metabolite repression. Nutrient starvation also causes the overall proteolysis rate in a cell to increase strongly, which makes sense for a cell that lacks nutrients but posses proteins, that under starvation conditions are not needed or needed only in smaller amounts. In expression strategies which allow high expression on media containing high glucose and ammonium concentrations reduced proteolysis has been reported. Several constitutive glycolytic promoters (gpd and pkiA) are highly expressed under these conditions and can also be used to drive (heterologous) gene expression in continuous fermentations. The type of nutrient starvation imposed can influence different proteases to varying extent, which means that the importance of nutrient conditions in a given process depend on the type of proteolysis that is involved. Specific proteolysis may therefore be induced by conditions of substrate limitation which are frequently used in many large-scale fermentation processes.
[0044]The protease problem can nowadays be addressed in part by one or more of the above strategies. However, the residual proteolytic activity of yet unidentified proteolytic enzymes still constitutes a major problem in the art. In order to further reduce the level of unwanted proteolysis, there is a great need in the art to identify novel proteases responsible for degradation of homologously and heterologously expressed proteins. This invention provides such novel protease gene sequences encoding novel proteases. Once the primary sequence of a novel protease gene is known, one or more of the above recombinant DNA strategies may be employed to produce (knock-out) mutants with reduced proteolytic activity.
[0045]Despite the widespread applications of proteases in a great number of industrial processes, current enzymes also have significant shortcomings with respect to at least one of the following properties.
[0046]When added to animal feed, current proteases are not sufficiently resistant to digestive enzymes present in the gastrointestinal (GI) tract of e.g. pigs and poultry.
[0047]With respect to another aspect, the currently available enzymes are not sufficiently resistant to specific (high) temperatures and (high) pressure conditions that are applied during extrusion or pelleting operations.
[0048]Also, the current enzymes are not sufficiently active in a pH range of 3-7, conditions prevailing in many food, beverage products as well as in the GI tract of most animals.
[0049]According to yet another aspect the specificity of the currently available proteases is very limited which results in the inability of the existing enzymes to degrade or to dissolve certain "protease resistant" proteins thus resulting in low peptide or amino acid yields. Moreover proteases with new specificities allow the synthesis of new peptides.
[0050]Yet another drawback of the currently available enzymes is their low specific activity.
[0051]It is therefore clear that for a large number of applications a strong desire exists for proteases that are more resistant to digestive enzymes, high temperature and/or pressure and which exhibit novel specificities regarding their sites of hydrolysis. The present invention provides such enzymes.
OBJECT OF THE INVENTION
[0052]It is an object of the invention to provide novel polynucleotides encoding novel proteases. A further object is to provide naturally and recombinantly produced proteases as well as recombinant strains producing these. Such strains may also be used to produce classical fermentation products faster or with higher yields. Yet another object of the invention is to provide a filamentous fungus strain defective in producing a protease according to the invention. Such strains may be used for a more efficient production of heterologous or homologous proteins. Also antibodies and fusion polypeptides are part of the invention as well as methods of making and using the polynucleotides and polypeptides according to the invention.
SUMMARY OF THE INVENTION
[0053]The invention provides for novel polynucleotides encoding novel proteases.
[0054]More in particular, the invention provides for polynucleotides having a nucleotide sequence that hybridises (preferably under highly stringent conditions) to a sequence according to a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or to a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114. Consequently, the invention provides nucleic acids that are about 60%, preferably 65%, more preferably 70%, even more preferably 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% homologous to the sequences according to a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114.
[0055]In a more preferred embodiment the invention provides for such an isolated polynucleotide obtainable from a filamentous fungus, preferably Aspergilli, in particular A. niger is preferred.
[0056]In one embodiment, the invention provides for an isolated polynucleotide comprising a nucleic acid sequence encoding a polypeptide with an amino acid sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 or functional equivalents thereof.
[0057]In a further preferred embodiment, the invention provides an isolated polynucleotide encoding at least one functional domain of a polypeptide according to a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 or functional equivalents thereof.
[0058]In a preferred embodiment the invention provides a protease gene according to a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57. In another aspect the invention provides a polynucleotide, preferably a cDNA encoding an A. niger protease selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 or variants or fragments of that polypeptide. In a preferred embodiment the cDNA has a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 or functional equivalents thereof.
[0059]A genomic clone encoding a polypeptide according to the invention may also be obtained by selecting suitable probes to specifically amplify a genomic region corresponding to any of the sequences according to SEQ ID NO: 1 to SEQ ID NO: 57 or fragments thereof, hybridising that probe under suitable conditions to genomic DNA obtained from a suitable organism, such as Aspergillus, e.g. A. niger, amplifying the desired fragment e.g. by PCR (polymerase chain reaction) followed by purifying and cloning of the amplified fragment.
[0060]In an even further preferred embodiment, the invention provides for a polynucleotide comprising the coding sequence of the genomic polynucleotides according to the invention, preferred is a polynucleotide sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114.
[0061]In another preferred embodiment, the invention provides a cDNA obtainable by cloning and expressing a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 into a suitable host organism, such as A. niger.
[0062]A polypeptide according to the invention may also be obtained by cloning and expressing a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 into a suitable host organism, such as A. niger.
[0063]The invention also relates to vectors comprising a polynucleotide sequence according to the invention and primers, probes and fragments that may be used to amplify or detect the DNA according to the invention.
[0064]In a further preferred embodiment, a vector is provided wherein the polynucleotide sequence according to the invention is functionally linked with regulatory sequences suitable for expression of the encoded amino acid sequence in a suitable host cell, such as A. niger or A. oryzea. The invention also provides methods for preparing polynucleotides and vectors according to the invention.
[0065]The invention also relates to recombinantly produced host cells that contain heterologous or homologous polynucleotides according to the invention.
[0066]In one embodiment, the invention provides recombinant host cells wherein the expression of a protease according to the invention is significantly reduced or wherein the activity of the protease is reduced or wherein the protease is even inactivated. Such recombinants are especially useful for the expression of homologous or heterologous proteins.
[0067]In another embodiment, the invention provides recombinant host cells wherein the expression of a protease according to the invention is significantly increased or wherein the activity of the protease is increased. Such recombinants are especially useful for the expression of homologous or heterologous proteins where maturation is seriously hampered in case the required proteolytic cleavage becomes the rate limiting step.
[0068]In another embodiment the invention provides for a recombinantly produced host cell that contains heterologous or homologous DNA according to the invention, preferably DNA encoding proteins bearing signal sequences and wherein the cell is capable of producing a functional protease according to the invention, preferably a cell capable of over-expressing the protease according to the invention, for example an Aspergillus strain comprising an increased copy number of a gene or cDNA according to the invention.
[0069]In another embodiment the invention provides for a recombinantly produced host cell that contains heterologous or homologous DNA according to the invention and wherein the cell is capable of secreting a functional protease according to the invention, preferably a cell capable of over-expressing and secreting the protease according to the invention, for example an Aspergillus strain comprising an increased copy number of a gene or cDNA according to the invention.
[0070]In yet another aspect of the invention, a purified polypeptide is provided. The polypeptides according to the invention include the polypeptides encoded by the polynucleotides according to the invention. Especially preferred is a polypeptide according to a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 or functional equivalents thereof.
[0071]The invention also provides for antibodies reactive with a polypeptide according to the invention. These antibodies may be polyclonal, yet especially preferred are monoclonal antibodies. Such antibodies are particularly useful for purifying the polypeptides according to the invention.
[0072]Fusion proteins comprising a polypeptide according to the invention are also within the scope of the invention. The invention also provides methods of making the polypeptides according to the invention.
[0073]The invention further relates to a method for diagnosing aspergillosis either by detecting the presence of a polypeptide according to the invention or functional equivalents thereof, or by detecting the presence of a DNA according to the invention or fragments or functional equivalents thereof.
[0074]The invention also relates to the use of the protease according to the invention in an industrial process as described herein
DETAILED DESCRIPTION OF THE INVENTION
Polynucleotides
[0075]The present invention provides polynucleotides encoding proteases having an amino acid sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 or functional equivalents thereof. The sequence of these genes was determined by sequencing a genomic clone obtained from Aspergillus niger. The invention provides polynucleotide sequences comprising the gene encoding these proteases as well as their complete cDNA sequence and its coding sequence. Accordingly, the invention relates to an isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 or functional equivalents thereof.
[0076]More in particular, the invention relates to an isolated polynucleotide hybridisable under stringent conditions to a polynucleotide selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 preferably under highly stringent conditions. Advantageously, such polynucleotides may be obtained from filamentous fungi, in particular from Aspergillus niger. More specifically, the invention relates to an isolated polynucleotide having a nucleotide sequence according to a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114.
[0077]The invention also relates to an isolated polynucleotide encoding at least one functional domain of a polypeptide according to a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 or functional equivalents thereof.
[0078]As used herein, the terms "gene" and "recombinant gene" refer to nucleic acid molecules which may be isolated from chromosomal DNA, which include an open reading frame encoding a protein, e.g. an A. niger protease. A gene may include coding sequences, non-coding sequences, introns and regulatory sequences. Moreover, a gene refers to an isolated nucleic acid molecule as defined herein.
[0079]A nucleic acid molecule of the present invention, such as a nucleic acid molecule having the nucleotide sequence of a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 or a functional equivalent thereof, can be isolated using standard molecular biology techniques and the sequence information provided herein. For example, using all or portion of the nucleic acid sequence of a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or the nucleotide sequence of a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 as a hybridization probe, nucleic acid molecules according to the invention can be isolated using standard hybridization and cloning techniques (e.g., as described in Sambrook, J., Fritsh, E. F., and Maniatis, T. Molecular Cloning: A Laboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989).
[0080]Moreover, a nucleic acid molecule encompassing all or a portion of a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 can be isolated by the polymerase chain reaction (PCR) using synthetic oligonucleotide primers designed based upon the sequence information contained in a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114.
[0081]A nucleic acid of the invention can be amplified using cDNA, mRNA or alternatively, genomic DNA, as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques. The nucleic acid so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis.
[0082]Furthermore, oligonucleotides corresponding to or hybridisable to nucleotide sequences according to the invention can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer.
[0083]In a preferred embodiment, an isolated nucleic acid molecule of the invention comprises the nucleotide sequence shown in a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114. The sequence of a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 corresponds to the coding region of the A. niger protease cDNA. This cDNA comprises sequences encoding the A. niger protease polypeptide according to a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171.
[0084]In another preferred embodiment, an isolated nucleic acid molecule of the invention comprises a nucleic acid molecule which is a complement of the nucleotide sequence shown in a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 or a functional equivalent of these nucleotide sequences.
[0085]A nucleic acid molecule which is complementary to another nucleotide sequence is one which is sufficiently complementary to the other nucleotide sequence such that it can hybridize to the other nucleotide sequence thereby forming a stable duplex.
[0086]One aspect of the invention pertains to isolated nucleic acid molecules that encode a polypeptide of the invention or a functional equivalent thereof such as a biologically active fragment or domain, as well as nucleic acid molecules sufficient for use as hybridisation probes to identify nucleic acid molecules encoding a polypeptide of the invention and fragments of such nucleic acid molecules suitable for use as PCR primers for the amplification or mutation of nucleic acid molecules.
[0087]An "isolated polynucleotide" or "isolated nucleic acid" is a DNA or RNA that is not immediately contiguous with both of the coding sequences with which it is immediately contiguous (one on the 5' end and one on the 3' end) in the naturally occurring genome of the organism from which it is derived. Thus, in one embodiment, an isolated nucleic acid includes some or all of the 5' non-coding (e.g., promotor) sequences that are immediately contiguous to the coding sequence. The term therefore includes, for example, a recombinant DNA that is incorporated into a vector, into an autonomously replicating plasmid or virus, or into the genomic DNA of a prokaryote or eukaryote, or which exists as a separate molecule (e.g., a cDNA or a genomic DNA fragment produced by PCR or restriction endonuclease treatment) independent of other sequences. It also includes a recombinant DNA that is part of a hybrid gene encoding an additional polypeptide that is substantially free of cellular material, viral material, or culture medium (when produced by recombinant DNA techniques), or chemical precursors or other chemicals (when chemically synthesized). Moreover, an "isolated nucleic acid fragment" is a nucleic acid fragment that is not naturally occurring as a fragment and would not be found in the natural state.
[0088]As used herein, the terms "polynucleotide" or "nucleic acid molecule" are intended to include DNA molecules (e.g., cDNA or genomic DNA) and RNA molecules (e.g., mRNA) and analogs of the DNA or RNA generated using nucleotide analogs. The nucleic acid molecule can be single-stranded or double-stranded, but preferably is double-stranded DNA. The nucleic acid may be synthesized using oligonucleotide analogs or derivatives (e.g., inosine or phosphorothioate nucleotides). Such oligonucleotides can be used, for example, to prepare nucleic acids that have altered base-pairing abilities or increased resistance to nucleases.
[0089]Another embodiment of the invention provides an isolated nucleic acid molecule which is antisense to a protease nucleic acid molecule, e.g., the coding strand of a protease nucleic acid molecule. Also included within the scope of the invention are the complement strands of the nucleic acid molecules described herein.
Sequencing Errors
[0090]The sequence information as provided herein should not be so narrowly construed as to require inclusion of erroneously identified bases. The specific sequences disclosed herein can be readily used to isolate the complete gene from filamentous fungi, in particular A. niger which in turn can easily be subjected to further sequence analyses thereby identifying sequencing errors.
[0091]Unless otherwise indicated, all nucleotide sequences determined by sequencing a DNA molecule herein were determined using an automated DNA sequencer and all amino acid sequences of polypeptides encoded by DNA molecules determined herein were predicted by translation of a DNA sequence determined as above. Therefore, as is known in the art for any DNA sequence determined by this automated approach, any nucleotide sequence determined herein may contain some errors. Nucleotide sequences determined by automation are typically at least about 90% identical, more typically at least about 95% to at least about 99.9% identical to the actual nucleotide sequence of the sequenced DNA molecule. The actual sequence can be more precisely determined by other approaches including manual DNA sequencing methods well known in the art. As is also known in the art, a single insertion or deletion in a determined nucleotide sequence compared to the actual sequence will cause a frame shift in translation of the nucleotide sequence such that the predicted amino acid sequence encoded by a determined nucleotide sequence will be completely different from the amino acid sequence actually encoded by the sequenced DNA molecule, beginning at the point of such an insertion or deletion.
[0092]The person skilled in the art is capable of identifying such erroneously identified bases and knows how to correct for such errors.
Nucleic Acid Fragments, Probes and Primers
[0093]A nucleic acid molecule according to the invention may comprise only a portion or a fragment of the nucleic acid sequence shown in a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114, for example a fragment which can be used as a probe or primer or a fragment encoding a portion of a protease protein. The nucleotide sequence determined from the cloning of the protease gene and cDNA allows for the generation of probes and primers designed for use in identifying and/or cloning other protease family members, as well as protease homologues from other species. The probe/primer typically comprises substantially purified oligonucleotide which typically comprises a region of nucleotide sequence that hybridizes preferably under highly stringent conditions to at least about 12 or 15, preferably about 18 or 20, preferably about 22 or 25, more preferably about 30, 35, 40, 45, 50, 55, 60, 65, or 75 or more consecutive nucleotides of a nucleotide sequence shown in a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 or of a functional equivalent thereof.
[0094]Probes based on the protease nucleotide sequences can be used to detect transcripts or genomic protease sequences encoding the same or homologous proteins for instance in other organisms. In preferred embodiments, the probe further comprises a label group attached thereto, e.g., the label group can be a radioisotope, a fluorescent compound, an enzyme, or an enzyme cofactor. Such probes can also be used as part of a diagnostic test kit for identifying cells which express a protease protein.
Identity & Homology
[0095]The terms "homology" or "percent identity" are used interchangeably herein. For the purpose of this invention, it is defined here that in order to determine the percent identity of two amino acid sequences or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity=number of identical positions/total number of positions (i.e. overlapping positions)×100).
[0096]Preferably, the two sequences are the same length.
[0097]The skilled person will be aware of the fact that several different computer programs are available to determine the homology between two sequences. For instance, a comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch (J. Mol. Biol. (48):444-453 (1970)) algorithm which has been incorporated into the GAP program in the GCG software package (available at http://www.gcg.com), using either a Blossom 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. The skilled person will appreciate that all these different parameters will yield slightly different results but that the overall percentage identity of two sequences is not significantly altered when using different algorithms.
[0098]In yet another embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (available at http://www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. In another embodiment, the percent identity two amino acid or nucleotide sequence is determined using the algorithm of E. Meyers and W. Miller (CABIOS, 4:11-17 (1989) which has been incorporated into the ALIGN program (version 2.0) (available at http://vega/igh.cnrs.fr/bin/align-guess.cgi), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
[0099]The nucleic acid and protein sequences of the present invention can further be used as a "query sequence" to perform a search against public databases to, for example, identify other family members or related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10. BLAST nucleotide searches can be performed with the NBLAST program, score=100, wordlength=12 to obtain nucleotide sequences homologous to protease nucleic acid molecules of the invention. BLAST protein searches can be performed with the XBLAST program, score=50, wordlength=3 to obtain amino acid sequences homologous to protease protein molecules of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res. 25(17):3389-3402. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See http://www.ncbi.nlm.nih.gov.
Hybridisation
[0100]As used herein, the term "hybridizing" is intended to describe conditions for hybridization and washing under which nucleotide sequences at least about 50%, at least about 60%, at least about 70%, more preferably at least about 80%, even more preferably at least about 85% to 90%, more preferably at least 95% homologous to each other typically remain hybridized to each other.
[0101]A preferred, non-limiting example of such hybridization conditions are hybridization in 6× sodium chloride/sodium citrate (SSC) at about 45° C., followed by one or more washes in 1×SSC, 0.1% SDS at 50° C., preferably at 55° C., preferably at 60° C. and even more preferably at 65° C.
[0102]Highly stringent conditions include, for example, hybridizing at 68° C. in 5×SSC/5×Denhardt's solution/1.0% SDS and washing in 0.2×SSC/0.1% SDS at room temperature. Alternatively washing may be performed at 42° C.
[0103]The skilled artisan will know which conditions to apply for stringent and highly stringent hybridisation conditions. Additional guidance regarding such conditions is readily available in the art, for example, in Sambrook et al., 1989, Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, N.Y.; and Ausubel et al. (eds.), 1995, Current Protocols in Molecular Biology, (John Wiley & Sons, N.Y.).
[0104]Of course, a polynucleotide which hybridizes only to a poly A sequence (such as the 3' terminal poly(A) tract of mRNAs), or to a complementary stretch of T (or U) resides, would not be included in a polynucleotide of the invention used to specifically hybridize to a portion of a nucleic acid of the invention, since such a polynucleotide would hybridize to any nucleic acid molecule contain a poly (A) stretch or the complement thereof (e.g., practically any double-stranded cDNA clone).
Obtaining Full Length DNA from Other Organisms
[0105]In a typical approach, cDNA libraries constructed from other organisms, e.g. filamentous fungi, in particular from the species Aspergillus can be screened.
[0106]For example, Aspergillus strains can be screened for homologous protease polynucleotides by Northern blot analysis. Upon detection of transcripts homologous to polynucleotides according to the invention, cDNA libraries can be constructed from RNA isolated from the appropriate strain, utilizing standard techniques well known to those of skill in the art. Alternatively, a total genomic DNA library can be screened using a probe hybridisable to a protease polynucleotide according to the invention.
[0107]Homologous gene sequences can be isolated, for example, by performing PCR using two oligonucleotide primers or two degenerate oligonucleotide primer pools designed on the basis of nucleotide sequences as taught herein.
[0108]The template for the reaction can be cDNA obtained by reverse transcription of mRNA prepared from strains known or suspected to express a polynucleotide according to the invention. The PCR product can be subcloned and sequenced to ensure that the amplified sequences represent the sequences of a new protease nucleic acid sequence, or a functional equivalent thereof.
[0109]The PCR fragment can then be used to isolate a full length cDNA clone by a variety of known methods. For example, the amplified fragment can be labeled and used to screen a bacteriophage or cosmid cDNA library. Alternatively, the labeled fragment can be used to screen a genomic library.
[0110]PCR technology also can be used to isolate full length cDNA sequences from other organisms. For example, RNA can be isolated, following standard procedures, from an appropriate cellular or tissue source. A reverse transcription reaction can be performed on the RNA using an oligonucleotide primer specific for the most 5' end of the amplified fragment for the priming of first strand synthesis.
[0111]The resulting RNA/DNA hybrid can then be "tailed" (e.g., with guanines) using a standard terminal transferase reaction, the hybrid can be digested with RNase H, and second strand synthesis can then be primed (e.g., with a poly-C primer). Thus, cDNA sequences upstream of the amplified fragment can easily be isolated. For a review of useful cloning strategies, see e.g., Sambrook et al., supra; and Ausubel et al., supra.
Vectors
[0112]Another aspect of the invention pertains to vectors, preferably expression vectors, containing a nucleic acid encoding a protease protein or a functional equivalent thereof. As used herein, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid", which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "expression vectors". In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. The terms "plasmid" and "vector" can be used interchangeably herein as the plasmid is the most commonly used form of vector. However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.
[0113]The recombinant expression vectors of the invention comprise a nucleic acid of the invention in a form suitable for expression of the nucleic acid in a host cell, which means that the recombinant expression vector includes one or more regulatory sequences, selected on the basis of the host cells to be used for expression, which is operatively linked to the nucleic acid sequence to be expressed. Within a recombinant expression vector, "operatively linked" is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner which allows for expression of the nucleotide sequence (e.g., in an in vitro transcription/translation system or in a host cell when the vector is introduced into the host cell). The term "regulatory sequence" is intended to include promoters, enhancers and other expression control elements (e.g., polyadenylation signal). Such regulatory sequences are described, for example, in Goeddel; Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990). Regulatory sequences include those which direct constitutive expression of a nucleotide sequence in many types of host cells and those which direct expression of the nucleotide sequence only in a certain host cell (e.g. tissue-specific regulatory sequences). It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, etc. The expression vectors of the invention can be introduced into host cells to thereby produce proteins or peptides, encoded by nucleic acids as described herein (e.g. protease proteins, mutant forms of protease proteins, fragments, variants or functional equivalents thereof, fusion proteins, etc.).
[0114]The recombinant expression vectors of the invention can be designed for expression of protease proteins in prokaryotic or eukaryotic cells. For example, protease proteins can be expressed in bacterial cells such as E. coli, insect cells (using baculovirus expression vectors) yeast cells or mammalian cells. Suitable host cells are discussed further in Goeddel, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990). Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.
[0115]Expression vectors useful in the present invention include chromosomal-, episomal- and virus-derived vectors e.g., vectors derived from bacterial plasmids, bacteriophage, yeast episome, yeast chromosomal elements, viruses such as baculoviruses, papova viruses, vaccinia viruses, adenoviruses, fowl pox viruses, pseudorabies viruses and retroviruses, and vectors derived from combinations thereof, such as those derived from plasmid and bacteriophage genetic elements, such as cosmids and phagemids.
[0116]The DNA insert should be operatively linked to an appropriate promoter, such as the phage lambda PL promoter, the E. coli lac, trp and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few. Other suitable promoters will be known to the skilled person. In a specific embodiment, promoters are preferred that are capable of directing a high expression level of proteases in filamentous fungi. Such promoters are known in the art. The expression constructs may contain sites for transcription initiation, termination, and, in the transcribed region, a ribosome binding site for translation. The coding portion of the mature transcripts expressed by the constructs will include a translation initiating AUG at the beginning and a termination codon appropriately positioned at the end of the polypeptide to be translated.
[0117]Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques. As used herein, the terms "transformation" and "transfection" are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co-precipitation, DEAE-dextran-mediated transfection, transduction, infection, lipofection, cationic lipid mediated transfection or electroporation. Suitable methods for transforming or transfecting host cells can be found in Sambrook, et al. (Molecular Cloning: A Laboratory Manual, 2nd, ed. Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989), Davis et al., Basic Methods in Molecular Biology (1986) and other laboratory manuals.
[0118]For stable transfection of mammalian cells, it is known that, depending upon the expression vector and transfection technique used, only a small fraction of cells may integrate the foreign DNA into their genome. In order to identify and select these integrants, a gene that encodes a selectable marker (e.g., resistance to antibiotics) is generally introduced into the host cells along with the gene of interest. Preferred selectable markers include those which confer resistance to drugs, such as G418, hygromycin and methatrexate. Nucleic acid encoding a selectable marker can be introduced into a host cell on the same vector as that encoding a protease protein or can be introduced on a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by drug selection (e.g. cells that have incorporated the selectable marker gene will survive, while the other cells die).
[0119]Expression of proteins in prokaryotes is often carried out in E. coli with vectors containing constitutive or inducible promoters directing the expression of either fusion or non-fusion proteins. Fusion vectors add a number of amino acids to a protein encoded therein, e.g. to the amino terminus of the recombinant protein. Such fusion vectors typically serve three purposes: 1) to increase expression of recombinant protein; 2) to increase the solubility of the recombinant protein; and 3) to aid in the purification of the recombinant protein by acting as a ligand in affinity purification. Often, in fusion expression vectors, a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant protein to enable separation of the recombinant protein from the fusion moiety subsequent to purification of the fusion protein. Such enzymes, and their cognate recognation sequences, include Factor Xa, thrombin and enterokinase.
[0120]As indicated, the expression vectors will preferably contain selectable markers. Such markers include dihydrofolate reductase or neomycin resistance for eukarotic cell culture and tetracyline or ampicilling resistance for culturing in E. coli and other bacteria. Representative examples of appropriate host include bacterial cells, such as E. coli, Streptomyces and Salmonella typhimurium; fungal cells, such as yeast; insect cells such as Drosophila S2 and Spodoptera Sf9; animal cells such as CHO, COS and Bowes melanoma; and plant cells. Appropriate culture mediums and conditions for the above-described host cells are known in the art.
[0121]Among vectors preferred for use in bacteria are pQE70, pQE60 and PQE-9, available from Qiagen; pBS vectors, Phagescript vectors, Bluescript vectors, pNH8A, pNH16A, pNH18A, pNH46A, available from Stratagene; and ptrc99a, pKK223-3, pKK233-3, pDR540, pRIT5 available from Pharmacia. Among preferred eukaryotic vectors are PWLNEO, pSV2CAT, pOG44, pZT1 and pSG available from Stratagene; and pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Other suitable vectors will be readily apparent to the skilled artisan.
[0122]Among known bacterial promotors for use in the present invention include E. coli lacI and lacZ promoters, the T3 and T7 promoters, the gpt promoter, the lambda PR, PL promoters and the trp promoter, the HSV thymidine kinase promoter, the early and late SV40 promoters, the promoters of retroviral LTRs, such as those of the Rous sarcoma virus ("RSV"), and metallothionein promoters, such as the mouse metallothionein-I promoter.
[0123]Transcription of the DNA encoding the polypeptides of the present invention by higher eukaryotes may be increased by inserting an enhancer sequence into the vector. Enhancers are cis-acting elements of DNA, usually about from 10 to 300 bp that act to increase transcriptional activity of a promoter in a given host cell-type. Examples of enhancers include the SV40 enhancer, which is located on the late side of the replication origin at bp 100 to 270, the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.
[0124]For secretion of the translated protein into the lumen of the endoplasmic reticulum, into the periplasmic space or into the extracellular environment, appropriate secretation signal may be incorporated into the expressed polypeptide. The signals may be endogenous to the polypeptide or they may be heterologous signals.
[0125]The polypeptide may be expressed in a modified form, such as a fusion protein, and may include not only secretion signals but also additional heterologous functional regions. Thus, for instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence in the host cell, during purification or during subsequent handling and storage. Also, peptide moieties may be added to the polypeptide to facilitate purification.
Polypeptides According to the Invention
[0126]The invention provides an isolated polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171, an amino acid sequence obtainable by expressing a polynucleotide according to the invention or in a preferred embodiment of a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 in an appropriate host, as well as an amino acid sequence obtainable by expressing a polynucleotide sequences selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 in an appropriate host. Also, a peptide or polypeptide comprising a functional equivalent of the above polypeptides is comprised within the present invention. The above polypeptides are collectively comprised in the term "polypeptides according to the invention"
[0127]The terms "peptide" and "oligopeptide" are considered synonymous (as is commonly recognized) and each term can be used interchangeably as the context requires to indicate a chain of at least two amino acids coupled by peptidyl linkages. The word "polypeptide" is used herein for chains containing more than seven amino acid residues. All oligopeptide and polypeptide formulas or sequences herein are written from left to right and in the direction from amino terminus to carboxy terminus. The one-letter code of amino acids used herein is commonly known in the art and can be found in Sambrook, et al. (Molecular Cloning: A Laboratory Manual, 2nd, ed. Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989)
[0128]By "isolated" polypeptide or protein is intended a polypeptide or protein removed from its native environment. For example, recombinantly produced polypeptides and proteins expressed in host cells are considered isolated for purpose of the invention as are native or recombinant polypeptides which have been substantially purified by any suitable technique such as, for example, the single-step purification method disclosed in Smith and Johnson, Gene 67:31-40 (1988).
[0129]The protease according to the invention can be recovered and purified from recombinant cell cultures by well-known methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. For analytical purposes most preferably, high performance liquid chromatography ("HPLC") is employed for purification.
[0130]Polypeptides of the present invention include naturally purified products, products of chemical synthetic procedures, and products produced by recombinant techniques from a prokaryotic or eukaryotic host, including, for example, bacterial, yeast, higher plant, insect and mammalian cells. Depending upon the host employed in a recombinant production procedure, the polypeptides of the present invention may be glycosylated or may be non-glycosylated. In addition, polypeptides of the invention may also include an initial modified methionine residue, in some cases as a result of host-mediated processes.
[0131]Moreover, a protein according to the invention may be a precursor protein such as a zymogen, a hybrid protein, a protein obtained as a pro sequence or pre-pro sequence, or any other type of immature form.
Protein Fragments
[0132]The invention also features biologically active fragments of the polypeptides according to the invention.
[0133]Biologically active fragments of a polypeptide of the invention include polypeptides comprising amino acid sequences sufficiently identical to or derived from the amino acid sequence of the protease protein (e.g., the amino acid sequence of a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171), which include fewer amino acids than the full length protein, and exhibit at least one biological activity of the corresponding full-length protein. Typically, biologically active fragments comprise a domain or motif with at least one activity of the protease protein. A biologically active fragment of a protein of the invention can be a polypeptide which is, for example, 10, 25, 50, 100 or more amino acids in length. Moreover, other biologically active portions, in which other regions of the protein are deleted, can be prepared by recombinant techniques and evaluated for one or more of the biological activities of the native form of a polypeptide of the invention.
[0134]The invention also features nucleic acid fragments which encode the above biologically active fragments of the protease protein.
Fusion Proteins
[0135]The proteins of the present invention or functional equivalents thereof, e.g., biologically active portions thereof, can be operatively linked to a non-protease polypeptide (e.g., heterologous amino acid sequences) to form fusion proteins. As used herein, a protease "chimeric protein" or "fusion protein" comprises a protease polypeptide operatively linked to a non-protease polypeptide. A "protease polypeptide" refers to a polypeptide having an amino acid sequence corresponding to a polypeptide sequence according to the invention, whereas a "non-protease polypeptide" refers to a polypeptide having an amino acid sequence corresponding to a protein which is not substantially homologous to a protein according to the invention, e.g., a protein which is different from the protease protein and which is derived from the same or a different organism. Within a protease fusion protein the protease polypeptide can correspond to all or a portion of a protein according to the invention. In a preferred embodiment, a protease fusion protein comprises at least one biologically active fragment of a protein according to the invention. In another preferred embodiment, a protease fusion protein comprises at least two biologically active portions of a protein according to the invention. Within the fusion protein, the term "operatively linked" is intended to indicate that the protease polypeptide and the non-protease polypeptide are fused in-frame to each other. The non-protease polypeptide can be fused to the N-terminus or C-terminus of the protease polypeptide.
[0136]For example, in one embodiment, the fusion protein is a GST-protease fusion protein in which the protease sequences are fused to the C-terminus of the GST sequences.
[0137]Such fusion proteins can facilitate the purification of recombinant protease. In another embodiment, the fusion protein is a protease protein containing a heterologous signal sequence at its N-terminus. In certain host cells (e.g., mammalian and Yeast host cells), expression and/or secretion of protease can be increased through use of a heterologous signal sequence.
[0138]In another example, the gp67 secretory sequence of the baculovirus envelope protein can be used as a heterologous signal sequence (Current Protocols in Molecular Biology, Ausubel et al., eds., John Wiley & Sons, 1992). Other examples of eukaryotic heterologous signal sequences include the secretory sequences of melittin and human placental alkaline phosphatase (Stratagene; La Jolla, Calif.). In yet another example, useful prokaryotic heterologous signal sequences include the phoA secretory signal (Sambrook et al., supra) and the protein A secretory signal (Pharmacia Biotech; Piscataway, N.J.).
[0139]A signal sequence can be used to facilitate secretion and isolation of a protein or polypeptide of the invention. Signal sequences are typically characterized by a core of hydrophobic amino acids which are generally cleaved from the mature protein during secretion in one or more cleavage events. Such signal peptides contain processing sites that allow cleavage of the signal sequence from the mature proteins as they pass through the secretory pathway. The signal sequence directs secretion of the protein, such as from a eukaryotic host into which the expression vector is transformed, and the signal sequence is subsequently or concurrently cleaved. The protein can then be readily purified from the extracellular medium by art recognized methods. Alternatively, the signal sequence can be linked to the protein of interest using a sequence which facilitates purification, such as with a GST domain. Thus, for instance, the sequence encoding the polypeptide may be fused to a marker sequence, such as a sequence encoding a peptide, which facilitates purification of the fused polypeptide. In certain preferred embodiments of this aspect of the invention, the marker sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (Qiagen, Inc.), among others, many of which are commercially available. As described in Gentz et al, Proc. Natl. Acad. Sci. USA 86:821-824 (1989), for instance, hexa-histidine provides for convenient purification of the fusion protein. The HA tag is another peptide useful for purification which corresponds to an epitope derived of influenza hemaglutinin protein, which has been described by Wilson et al., Cell 37:767 (1984), for instance.
[0140]Preferably, a protease chimeric or fusion protein of the invention is produced by standard recombinant DNA techniques. For example, DNA fragments coding for the different polypeptide sequences are ligated together in-frame in accordance with conventional techniques, for example by employing blunt-ended or stagger-ended termini for ligation, restriction enzyme digestion to provide for appropriate termini, filling-in of cohesive ends as appropriate, alkaline phosphatase treatment to avoid undesirable joining, and enzymatic ligation. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers which give rise to complementary overhangs between two consecutive gene fragments which can subsequently be annealed and reamplified to generate a chimeric gene sequence (see, for example, Current Protocols in Molecular Biology, eds. Ausubel et al. John Wiley & Sons: 1992). Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g, a GST polypeptide). A protease-encoding nucleic acid can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the protease protein.
Functional Equivalents
[0141]The terms "functional equivalents" and "functional variants" are used interchangeably herein. Functional equivalents of a DNA according to the invention are isolated DNA fragments that encode a polypeptide that exhibits a particular function of an A. niger protease as defined herein. A functional equivalent of a polypeptide according to the invention is a polypeptide that exhibits at least one function of an A. niger protease as defined herein.
[0142]Functional protein or polypeptide equivalents may contain only conservative substitutions of one or more amino acids of a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 or substitutions, insertions or deletions of non-essential amino acids. Accordingly, a non-essential amino acid is a residue that can be altered in a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 without substantially altering the biological function. For example, amino acid residues that are conserved among the protease proteins of the present invention, are predicted to be particularly unamenable to alteration. Furthermore, amino acids conserved among the protease proteins according to the present invention and other proteases are not likely to be amenable to alteration.
[0143]The term "conservative substitution" is intended to mean that a substitution in which the amino acid residue is replaced with an amino acid residue having a similar side chain. These families are known in the art and include amino acids with basic side chains (e.g. lysine, arginine and hystidine), acidic side chains (e.g. aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagines, glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine tryptophan, histidine).
[0144]Functional nucleic acid equivalents may typically contain silent mutations or mutations that do not alter the biological function of encoded polypeptide. Accordingly, the invention provides nucleic acid molecules encoding protease proteins that contain changes in amino acid residues that are not essential for a particular biological activity. Such protease proteins differ in amino acid sequence from a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 yet retain at least one biological activity. In one embodiment the isolated nucleic acid molecule comprises a nucleotide sequence encoding a protein, wherein the protein comprises a substantially homologous amino acid sequence of at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more homologous to the amino acid sequence shown in a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171.
[0145]For example, guidance concerning how to make phenotypically silent amino acid substitutions is provided in Bowie, J. U. et al., Science 247:1306-1310 (1990) wherein the authors indicate that there are two main approaches for studying the tolerance of an amino acid sequence to change. The first method relies on the process of evolution, in which mutations are either accepted or rejected by natural selection. The second approach uses genetic engineering to introduce amino acid changes at specific positions of a cloned gene and selects or screens to identify sequences that maintain functionality. As the authors state, these studies have revealed that proteins are surprisingly tolerant of amino acid substitutions. The authors further indicate which changes are likely to be permissive at a certain position of the protein. For example, most buried amino acid residues require non-polar side chains, whereas few features of surface side chains are generally conserved. Other such phenotypically silent substitutions are described in Bowie et al, supra, and the references cited therein.
[0146]An isolated nucleic acid molecule encoding a protease protein homologous to the protein selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 can be created by introducing one or more nucleotide substitutions, additions or deletions into the coding nucleotide sequences according to a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 such that one or more amino acid substitutions, deletions or insertions are introduced into the encoded protein. Such mutations may be introduced by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis.
[0147]The term "functional equivalents" also encompasses orthologues of the A. niger protease protein. Orthologues of the A. niger protease protein are proteins that can be isolated from other strains or species and possess a similar or identical biological activity. Such orthologues can readily be identified as comprising an amino acid sequence that is substantially homologous to a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO:171.
[0148]As defined herein, the term "substantially homologous" refers to a first amino acid or nucleotide sequence which contains a sufficient or minimum number of identical or equivalent (e.g., with similar side chain) amino acids or nucleotides to a second amino acid or nucleotide sequence such that the first and the second amino acid or nucleotide sequences have a common domain. For example, amino acid or nucleotide sequences which contain a common domain having about 60%, preferably 65%, more preferably 70%, even more preferably 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity or more are defined herein as sufficiently identical.
[0149]Also, nucleic acids encoding other protease family members, which thus have a nucleotide sequence that differs from a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114, are within the scope of the invention. Moreover, nucleic acids encoding protease proteins from different species which thus have a nucleotide sequence which differs from a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 are within the scope of the invention.
[0150]Nucleic acid molecules corresponding to variants (e.g. natural allelic variants) and homologues of the protease DNA of the invention can be isolated based on their homology to the protease nucleic acids disclosed herein using the cDNAs disclosed herein or a suitable fragment thereof, as a hybridisation probe according to standard hybridisation techniques preferably under highly stringent hybridisation conditions.
[0151]In addition to naturally occurring allelic variants of the protease sequence, the skilled person will recognise that changes can be introduced by mutation into the nucleotide sequences of a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 thereby leading to changes in the amino acid sequence of the protease protein without substantially altering the function of the protease protein.
[0152]In another aspect of the invention, improved protease proteins are provided. Improved protease proteins are proteins wherein at least one biological activity is improved. Such proteins may be obtained by randomly introducing mutations along all or part of the protease coding sequence, such as by saturation mutagenesis, and the resulting mutants can be expressed recombinantly and screened for biological activity. For instance, the art provides for standard assays for measuring the enzymatic activity of proteases and thus improved proteins may easily be selected.
[0153]In a preferred embodiment the protease protein has an amino acid sequence according to a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171. In another embodiment, the protease polypeptide is substantially homologous to the amino acid sequence according to a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 and retains at least one biological activity of a polypeptide according to a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171, yet differs in amino acid sequence due to natural variation or mutagenesis as described above.
[0154]In a further preferred embodiment, the protease protein has an amino acid sequence encoded by an isolated nucleic acid fragment capable of hybridising to a nucleic acid according to a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57 or a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114, preferably under highly stringent hybridisation conditions.
[0155]Accordingly, the protease protein is a protein which comprises an amino acid sequence at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more homologous to the amino acid sequence shown in a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 and retains at least one functional activity of the polypeptide according to a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171.
[0156]Functional equivalents of a protein according to the invention can also be identified e.g. by screening combinatorial libraries of mutants, e.g. truncation mutants, of the protein of the invention for protease activity. In one embodiment, a variegated library of variants is generated by combinatorial mutagenesis at the nucleic acid level. A variegated library of variants can be produced by, for example, enzymatically ligating a mixture of synthetic oligonucleotides into gene sequences such that a degenerate set of potential protein sequences is expressible as individual polypeptides, or alternatively, as a set of larger fusion proteins (e.g., for phage display). There are a variety of methods that can be used to produce libraries of potential variants of the polypeptides of the invention from a degenerate oligonucleotide sequence. Methods for synthesizing degenerate oligonucleotides are known in the art (see, e.g., Narang (1983) Tetrahedron 39:3; Itakura et al. (1984) Annu. Rev. Biochem. 53:323; Itakura et al. (1984) Science 198:1056; Ike et al. (1983) Nucleic Acid Res. 11:477).
[0157]In addition, libraries of fragments of the coding sequence of a polypeptide of the invention can be used to generate a variegated population of polypeptides for screening a subsequent selection of variants. For example, a library of coding sequence fragments can be generated by treating a double stranded PCR fragment of the coding sequence of interest with a nuclease under conditions wherein nicking occurs only about once per molecule, denaturing the double stranded DNA, renaturing the DNA to form double stranded DNA which can include sense/antisense pairs from different nicked products, removing single stranded portions from reformed duplexes by treatment with S1 nuclease, and ligating the resulting fragment library into an expression vector. By this method, an expression library can be derived which encodes N-terminal and internal fragments of various sizes of the protein of interest.
[0158]Several techniques are known in the art for screening gene products of combinatorial libraries made by point mutations of truncation, and for screening cDNA libraries for gene products having a selected property. The most widely used techniques, which are amenable to high through-put analysis, for screening large gene libraries typically include cloning the gene library into replicable expression vectors, transforming appropriate cells with the resulting library of vectors, and expressing the combinatorial genes under conditions in which detection of a desired activity facilitates isolation of the vector encoding the gene whose product was detected. Recursive ensemble mutagenesis (REM), a technique which enhances the frequency of functional mutants in the libraries, can be used in combination with the screening assays to identify variants of a protein of the invention (Arkin and Yourvan (1992) Proc. Natl. Acad. Sci. USA 89:7811-7815; Delgrave et al. (1993) Protein Engineering 6(3):327-331).
[0159]In addition to the protease gene sequence shown in a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57, it will be apparent for the person skilled in the art that DNA sequence polymorphisms that may lead to changes in the amino acid sequence of the protease protein may exist within a given population. Such genetic polymorphisms may exist in cells from different populations or within a population due to natural allelic variation. Allelic variants may also include functional equivalents.
[0160]Fragments of a polynucleotide according to the invention may also comprise polynucleotides not encoding functional polypeptides. Such polynucleotides may function as probes or primers for a PCR reaction. Such polynucleotides may also be useful when it is desired to abolish the functional activity of a protease in a particular organism (knock-out mutants).
[0161]Nucleic acids according to the invention irrespective of whether they encode functional or non-functional polypeptides, can be used as hybridization probes or polymerase chain reaction (PCR) primers. Uses of the nucleic acid molecules of the present invention that do not encode a polypeptide having a protease activity include, inter alia, (1) isolating the gene encoding the protease protein, or allelic variants thereof from a cDNA library e.g. from other organisms than A. niger; (2) in situ hybridization (e.g. FISH) to metaphase chromosomal spreads to provide precise chromosomal location of the protease gene as described in Verma et al., Human Chromosomes: a Manual of Basic Techniques, Pergamon Press, New York (1988); (3) Northern blot analysis for detecting expression of protease mRNA in specific tissues and/or cells and 4) probes and primers that can be used as a diagnostic tool to analyse the presence of a nucleic acid hybridisable to the protease probe in a given biological (e.g. tissue) sample.
[0162]Also encompassed by the invention is a method of obtaining a functional equivalent of a protease gene or cDNA. Such a method entails obtaining a labelled probe that includes an isolated nucleic acid which encodes all or a portion of the sequence according to a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 or a variant thereof; screening a nucleic acid fragment library with the labelled probe under conditions that allow hybridisation of the probe to nucleic acid fragments in the library, thereby forming nucleic acid duplexes, and preparing a full-length gene sequence from the nucleic acid fragments in any labelled duplex to obtain a gene related to the protease gene.
[0163]In one embodiment, a protease nucleic acid of the invention is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more homologous to a nucleic acid sequence shown in a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 57, a sequence selected from the group consisting of SEQ ID NO: 58 to SEQ ID NO: 114 or the complement thereof.
[0164]In another preferred embodiment a protease polypeptide of the invention is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more homologous to the amino acid sequence shown in a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171.
Host Cells
[0165]In another embodiment, the invention features cells, e.g., transformed host cells or recombinant host cells that contain a nucleic acid encompassed by the invention. A "transformed cell" or "recombinant cell" is a cell into which (or into an ancestor of which) has been introduced, by means of recombinant DNA techniques, a nucleic acid according to the invention. Both prokaryotic and eukaryotic cells are included, e.g., bacteria, fungi, yeast, and the like, especially preferred are cells from filamentous fungi, in particular Aspergillus niger.
[0166]A host cell can be chosen that modulates the expression of the inserted sequences, or modifies and processes the gene product in a specific, desired fashion. Such modifications (e.g., glycosylation) and processing (e.g., cleavage) of protein products may facilitate optimal functioning of the protein.
[0167]Various host cells have characteristic and specific mechanisms for post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems familiar to those of skill in the art of molecular biology and/or microbiology can be chosen to ensure the desired and correct modification and processing of the foreign protein expressed. To this end, eukaryotic host cells that possess the cellular machinery for proper processing of the primary transcript, glycosylation, and phosphorylation of the gene product can be used. Such host cells are well known in the art.
[0168]Host cells also include, but are not limited to, mammalian cell lines such as CHO, VERO, BHK, HeLa, COS, MDCK, 293, 3T3, WI38, and choroid plexus cell lines.
[0169]If desired, the polypeptides according to the invention can be produced by a stably-transfected cell line. A number of vectors suitable for stable transfection of mammalian cells are available to the public, methods for constructing such cell lines are also publicly known, e.g., in Ausubel et al. (supra).
Antibodies
[0170]The invention further features antibodies, such as monoclonal or polyclonal antibodies, that specifically bind protease proteins according to the invention.
[0171]As used herein, the term "antibody" (Ab) or "monoclonal antibody" (Mab) is meant to include intact molecules as well as antibody fragments (such as, for example, Fab and F(ab')2 fragments) which are capable of specifically binding to protease protein. Fab and F(ab')2 fragments lack the Fc fragment of intact antibody, clear more rapidly from the circulation, and may have less non-specific tissue binding of an intact antibody (Wahl et al, J. Nucl. Med. 24:316-325 (1983)). Thus, these fragments are preferred.
[0172]The antibodies of the present invention may be prepared by any of a variety of methods. For example, cells expressing the protease protein or an antigenic fragment thereof can be administered to an animal in order to induce the production of sera containing polyclonal antibodies. In a preferred method, a preparation of protease protein is prepared and purified to render it substantially free of natural contaminants. Such a preparation is then introduced into an animal in order to produce polyclonal antisera of greater specific activity.
[0173]In the most preferred method, the antibodies of the present invention are monoclonal antibodies (or protease protein binding fragments thereof). Such monoclonal antibodies can be prepared using hybridoma technology (Kohler et al., Nature 256':495 (1975); Kohler et al., Eur. J. Immunol. 6:511 (1976); Hammerling et al., In: Monoclonal Antibodies and T-Cell Hybridomas, Elsevier, N.Y., pp. 563-681 (1981)). In general, such procedures involve immunizing an animal (preferably a mouse) with a protease protein antigen or, with a protease protein expressing cell. The splenocytes of such mice are extracted and fused with a suitable myeloma cell line. Any suitable myeloma cell line may be employed in accordance with the present invention; however, it is preferably to employ the parent myeloma cell line (SP2O), available from the American Type Culture Collection, Rockville, Md. After fusion, the resulting hybridoma cells are selectively maintained in HAT medium, and then cloned by limiting dilution as described by Wands et al. (Gastro-enterology 80.225-232 (1981)). The hybridoma cells obtained through such a selection are then assayed to identify clones which secrete antibodies capable of binding the protease protein antigen. In general, the polypeptides can be coupled to a carrier protein, such as KLH, as described in Ausubel et al., supra, mixed with an adjuvant, and injected into a host mammal.
[0174]In particular, various host animals can be immunized by injection of a polypeptide of interest. Examples of suitable host animals include rabbits, mice, guinea pigs, and rats. Various adjuvants can be used to increase the immunological response, depending on the host species, including but not limited to Freund's (complete and incomplete), adjuvant mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemocyanin, dinitrophenol, BCG (bacille Calmette-Guerin) and Corynebacterium parvum. Polyclonal antibodies are heterogeneous populations of antibody molecules derived from the sera of the immunized animals.
[0175]Such antibodies can be of any immunoglobulin class including IgG, IgM, IgE, IgA, IgD, and any subclass thereof. The hybridomas producing the mAbs of this invention can be cultivated in vitro or in vivo.
[0176]Once produced, polyclonal or monoclonal antibodies are tested for specific recognition of an protease polypeptide or functional equivalent thereof in an immunoassay, such as a Western blot or immunoprecipitation analysis using standard techniques, e.g., as described in Ausubel et al., supra. Antibodies that specifically bind to protease proteins or functional equivalents thereof are useful in the invention. For example, such antibodies can be used in an immunoassay to detect protease in pathogenic or non-pathogenic strains of Aspergillus (e.g., in Aspergillus extracts).
[0177]Preferably, antibodies of the invention are produced using fragments of the protease polypeptides that appear likely to be antigenic, by criteria such as high frequency of charged residues. For example, such fragments may be generated by standard techniques of PCR, and then cloned into the pGEX expression vector (Ausubel et al., supra). Fusion proteins may then be expressed in E. coli and purified using a glutathione agarose affinity matrix as described in Ausubel, et al., supra. If desired, several (e.g., two or three) fusions can be generated for each protein, and each fusion can be injected into at least two rabbits. Antisera can be raised by injections in a series, typically including at least three booster injections. Typically, the antisera are checked for their ability to immunoprecipitate a recombinant protease polypeptide or functional equivalents thereof whereas unrelated proteins may serve as a control for the specificity of the immune reaction.
[0178]Alternatively, techniques described for the production of single chain antibodies (U.S. Pat. Nos. 4,946,778 and 4,704,692) can be adapted to produce single chain antibodies against a protease polypeptide or functional equivalents thereof. Kits for generating and screening phage display libraries are commercially available e.g. from Pharmacia.
[0179]Additionally, examples of methods and reagents particularly amenable for use in generating and screening antibody display library can be found in, for example, U.S. Pat. No. 5,223,409; PCT Publication No. WO 92/18619; PCT Publication No. WO 91/17271; PCT Publication No. WO 20791; PCT Publication No. WO 92/20791; PCT Publication No. WO 92/15679; PCT Publication No. WO 93/01288; PCT Publication No. WO 92/01047; PCT Publication No. WO 92/09690; PCT Publication No. WO 90/02809; Fuchs et al. (1991) Bio/Technology 9:1370-1372; Hay et al. (1992) Hum. Antibod. Hybridomas 3:81-85; Huse et al. (1989) Science 246; 1275-1281; Griffiths et al. (1993) EMBO J. 12:725-734.
[0180]Polyclonal and monoclonal antibodies that specifically bind protease polypeptides of functional equivalents thereof can be used, for example, to detect expression of a protease gene or a functional equivalent thereof e.g. in another strain of Aspergillus.
[0181]For example, protease polypeptide can be readily detected in conventional immunoassays of Aspergillus cells or extracts. Examples of suitable assays include, without limitation, Western blotting, ELISAs, radioimmune assays, and the like.
[0182]By "specifically binds" is meant that an antibody recognizes and binds a particular antigen, e.g., a protease polypeptide, but does not substantially recognize and bind other unrelated molecules in a sample.
[0183]Antibodies can be purified, for example, by affinity chromatography methods in which the polypeptide antigen is immobilized on a resin.
[0184]An antibody directed against a polypeptide of the invention (e.g., monoclonal antibody) can be used to isolate the polypeptide by standard techniques, such as affinity chromatography or immunoprecipitation. Moreover, such an antibody can be used to detect the protein (e.g., in a cellular lysate or cell supernatant) in order to evaluate the abundance and pattern of expression of the polypeptide. The antibodies can also be used diagnostically to monitor protein levels in cells or tissue as part of a clinical testing procedure, e.g., to, for example, determine the efficacy of a given treatment regimen or in the diagnosis of Aspergillosis.
[0185]Detection can be facilitated by coupling the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and acquorin, and examples of suitable radioactive materials include 125I, 131I, 35S or 3H.
[0186]Preferred epitopes encompassed by the antigenic peptide are regions that are located on the surface of the protein, e.g., hydrophilic regions. Hydrophobicity plots of the proteins of the invention can be used to identify hydrophilic regions.
[0187]The antigenic peptide of a protein of the invention comprises at least 7 (preferably 10, 15, 20, or 30) contiguous amino acid residues of the amino acid sequence of a sequence selected from the group consisting of SEQ ID NO: 115 to SEQ ID NO: 171 and encompasses an epitope of the protein such that an antibody raised against the peptide forms a specific immune complex with the protein.
[0188]Preferred epitopes encompassed by the antigenic peptide are regions of protease that are located on the surface of the protein, e.g., hydrophilic regions, hydrophobic regions, alpha regions, beta regions, coil regions, turn regions and flexible regions.
Immunoassays
[0189]Qualitative or quantitative determination of a polypeptide according to the present invention in a biological sample can occur using any art-known method. Antibody-based techniques provide special advantages for assaying specific polypeptide levels in a biological sample.
[0190]In these, the specific recognition is provided by the primary antibody (polyclonal or monoclonal) but the secondary detection system can utilize fluorescent, enzyme, or other conjugated secondary antibodies. As a result, an immunocomplex is obtained.
[0191]Accordingly, the invention provides a method for diagnosing whether a certain organism is infected with Aspergillus comprising the steps of: [0192]Isolating a biological sample from said organism suspected to be infected with Aspergillus, [0193]reacting said biological sample with an antibody according to the invention, [0194]determining whether immunocomplexes are formed.
[0195]Tissues can also be extracted, e.g., with urea and neutral detergent, for the liberation of protein for Western-blot or dot/slot assay. This technique can also be applied to body fluids.
[0196]Other antibody-based methods useful for detecting protease gene expression include immunoassays, such as the enzyme linked immunosorbent assay (ELISA) and the radioimmunoassay (RIA). For example, protease-specific monoclonal antibodies can be used both as an immunoabsorbent and as an enzyme-labeled probe to detect and quantify the protease protein. The amount of protease protein present in the sample can be calculated by reference to the amount present in a standard preparation using a linear regression computer algorithm. In another ELISA assay, two distinct specific monoclonal antibodies can be used to detect protease protein in a biological fluid. In this assay, one of the antibodies is used as the immuno-absorbent and the other as the enzyme-labeled probe.
[0197]The above techniques may be conducted essentially as a "one-step" or "two-step" assay. The "one-step" assay involves contacting protease protein with immobilized antibody and, without washing, contacting the mixture with the labeled antibody. The "two-step" assay involves washing before contacting the mixture with the labeled antibody. Other conventional methods may also be employed as suitable. It is usually desirable to immobilize one component of the assay system on a support, thereby allowing other components of the system to be brought into contact with the component and readily removed from the sample.
[0198]Suitable enzyme labels include, for example, those from the oxidase group, which catalyze the production of hydrogen peroxide by reacting with substrate. Activity of an oxidase label may be assayed by measuring the concentration of hydrogen peroxide formed by the enzyme-labelled antibody/substrate reaction.
[0199]Besides enzymes, other suitable labels include radioisotopes, such as iodine (125I, 121I), carbon (14C), sulphur (35S), tritium (3H), indium (112In), and technetium (99mTc), and fluorescent labels, such as fluorescein and rhodamine, and biotin.
[0200]Specific binding of a test compound to a protease polypeptide can be detected, for example, in vitro by reversibly or irreversibly immobilizing the protease polypeptide on a substrate, e.g., the surface of a well of a 96-well polystyrene microtitre plate. Methods for immobilizing polypeptides and other small molecules are well known in the art. For example, the microtitre plates can be coated with a protease polypeptide by adding the polypeptide in a solution (typically, at a concentration of 0.05 to 1 mg/ml in a volume of 1-100 ul) to each well, and incubating the plates at room temperature to 37° C. for 0.1 to 36 hours. Polypeptides that are not bound to the plate can be removed by shaking the excess solution from the plate, and then washing the plate (once or repeatedly) with water or a buffer. Typically, the polypeptide is contained in water or a buffer. The plate is then washed with a buffer that lacks the bound polypeptide. To block the free protein-binding sites on the plates, the plates are blocked with a protein that is unrelated to the bound polypeptide. For example, 300 ul of bovine serum albumin (BSA) at a concentration of 2 mg/ml in Tris-HCl is suitable. Suitable substrates include those substrates that contain a defined cross-linking chemistry (e.g., plastic substrates, such as polystyrene, styrene, or polypropylene substrates from Corning Costar Corp. (Cambridge, Mass.), for example). If desired, a beaded particle, e.g., beaded agarose or beaded sepharose, can be used as the substrate.
[0201]Binding of the test compound to the polypeptides according to the invention can be detected by any of a variety of art known methods. For example, a specific antibody can be used in an immunoassay. If desired, the antibody can be labeled (e.g., fluorescently or with a radioisotope) and detected directly (see, e.g., West and McMahon, J. Cell Biol. 74:264, 1977). Alternatively, a second antibody can be used for detection (e.g., a labeled antibody that binds the Fc portion of an anti-AN97 antibody). In an alternative detection method, the protease polypeptide is labeled, and the label is detected (e.g., by labeling a protease polypeptide with a radioisotope, fluorophore, chromophore, or the like). In still another method, the protease polypeptide is produced as a fusion protein with a protein that can be detected optically, e.g., green fluorescent protein (which can be detected under UV light). In an alternative method, the protease polypeptide can be covalently attached to or fused with an enzyme having a detectable enzymatic activity, such as horse radish peroxidase, alkaline phosphatase, a-galactosidase, or glucose oxidase. Genes encoding all of these enzymes have been cloned and are readily available for use by those of skill in the art. If desired, the fusion protein can include an antigen, and such an antigen can be detected and measured with a polyclonal or monoclonal antibody using conventional methods. Suitable antigens include enzymes (e.g., horse radish peroxidase, alkaline phosphatase, and a-galactosidase) and non-enzymatic polypeptides (e.g., serum proteins, such as BSA and globulins, and milk proteins, such as caseins).
Epitopes, Antigens and Immunogens.
[0202]In another aspect, the invention provides a peptide or polypeptide comprising an epitope-bearing portion of a polypeptide of the invention. The epitope of this polypeptide portion is an immunogenic or antigenic epitope of a polypeptide of the invention. An "immunogenic epitope" is defined as a part of a protein that elicits an antibody response when the whole protein is the immunogen. These immunogenic epitopes are believed to be confined to a few loci on the molecule. On the other hand, a region of a protein molecule to which an antibody can bind is defined as an "antigenic epitope." The number of immunogenic epitopes of a protein generally is less than the number of antigenic epitopes. See, for instance, Geysen, H. M. et al., Proc. Natl. Acad. Sci. USA 81:3998-4002 (1984).
[0203]As to the selection of peptides or polypeptides bearing an antigenic epitope (i.e., that contain a region of a protein molecule to which an antibody can bind), it is well known in that art that relatively short synthetic peptides that mimic part of a protein sequence are routinely capable of eliciting an antiserum that reacts with the partially mimicked protein. See, for instance, Sutcliffe, J. G. et al., Science 219:660-666 (1984). Peptides capable of eliciting protein-reactive sera are frequently represented in the primary sequence of a protein, can be characterized by a set of simple chemical rules, and are confined neither to immunodominant regions of intact proteins (i.e., immunogenic epitopes) nor to the amino or carboxyl terminals. Peptides that are extremely hydrophobic and those of six or fewer residues generally are ineffective at inducing antibodies that bind to the mimicked protein; longer, soluble peptides, especially those containing proline residues, usually are effective. Sutcliffe et al., supra ?. For instance, 18 of 20 peptides designed according to these guidelines, containing 8-39 residues covering 75% of the sequence of the influenza virus hemagglutinin HAI polypeptide chain, induced antibodies that reacted with the HA1 protein or intact virus; and 12/12 peptides from the MuLV polymerase and 18/18 from the rabies glycoprotein induced antibodies that precipitated the respective proteins.
[0204]Antigenic epitope-bearing peptides and polypeptides of the invention are therefore useful to raise antibodies, including monoclonal antibodies, that bind specifically to a polypeptide of the invention. Thus, a high proportion of hybridomas obtained by fusion of spleen cells from donors immunized with an antigen epitope-bearing peptide generally secrete antibody reactive with the native protein. Sutcliffe et al., supra, at 663. The antibodies raised by antigenic epitope bearing peptides or polypeptides are useful to detect the mimicked protein, and antibodies to different peptides may be used for tracking the fate of various regions of a protein precursor which undergoes posttranslation processing. The peptides and anti-peptide antibodies may be used in a variety of qualitative or quantitative assays for the mimicked protein, for instance in competition assays since it has been shown that even short peptides (e.g., about 9 amino acids) can bind and displace the larger peptides in immunoprecipitation assays. See, for instance, Wilson, I. A. et al., Cell 37:767-778 at 777 (1984). The anti-peptide antibodies of the invention also are useful for purification of the mimicked protein, for instance, by adsorption chromatography using methods well known in the art.
[0205]Antigenic epitope-bearing peptides and polypeptides of the invention designed according to the above guidelines preferably contain a sequence of at least seven, more preferably at least nine and most preferably between about 15 to about 30 amino acids contained within the amino acid sequence of a polypeptide of the invention. However, peptides or polypeptides comprising a larger portion of an amino acid sequence of a polypeptide of the invention, containing about 30 to about 50 amino acids, or any length up to and including the entire amino acid sequence of a polypeptide of the invention, also are considered epitope-bearing peptides or polypeptides of the invention and also are useful for inducing antibodies that react with the mimicked protein. Preferably, the amino acid sequence of the epitope-bearing peptide is selected to provide substantial solubility in aqueous solvents (i.e., the sequence includes relatively hydrophilic residues and highly hydrophobic sequences are preferably avoided); and sequences containing proline residues are particularly preferred.
[0206]The epitope-bearing peptides and polypeptides of the invention may be produced by any conventional means for making peptides or polypeptides including recombinant means using nucleic acid molecules of the invention. For instance, a short epitope-bearing amino acid sequence may be fused to a larger polypeptide which acts as a carrier during recombinant production and purification, as well as during immunization to produce anti-peptide antibodies.
[0207]Epitope-bearing peptides also may be synthesized using known methods of chemical synthesis. For instance, Houghten has described a simple method for synthesis of large numbers of peptides, such as 10-20 mg of 248 different 13 residue peptides representing single amino acid variants of a segment of the HAI polypeptide which were prepared and characterized (by ELISA-type binding studies) in less than four weeks. Houghten, R. A., Proc. Natl. Acad. Sci. USA 82:5131-5135 (1985). This "Simultaneous Multiple Peptide Synthesis (SMPS)" process is further described in U.S. Pat. No. 4,631,211 to Houghten et al. (1986). In this procedure the individual resins for the solid-phase synthesis of various peptides are contained in separate solvent-permeable packets, enabling the optimal use of the many identical repetitive steps involved in solid-phase methods.
[0208]A manual procedure allows 500-1000 or more syntheses to be conducted simultaneously. Houghten et al., supra, at 5134.
[0209]Epitope-bearing peptides and polypeptides of the invention are used to induce antibodies according to methods well known in the art. See, for instance, Sutcliffe et al., supra; Wilson et al., supra; Chow, M. et al., Proc. Natl. Acad. Sci. USA 82:910-914; and Bittle, F. J. et al., J. Gen. Virol. 66:2347-2354 (1985).
[0210]Generally, animals may be immunized with free peptide; however, anti-peptide antibody titer may be boosted by coupling of the peptide to a macromolecular carrier, such as keyhole limpet hemocyanin (KLH) or tetanus toxoid. For instance, peptides containing cysteine may be coupled to carrier using a linker such as maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), while other peptides may be coupled to carrier using a more general linking agent such as glutaraldehyde.
[0211]Animals such as rabbits, rats and mice are immunized with either free or carrier coupled peptides, for instance, by intraperitoneal and/or intradermal injection of emulsions containing about 100 ug peptide or carrier protein and Freund's adjuvant. Several booster injections may be needed, for instance, at intervals of about two weeks, to provide a useful titer of anti-peptide antibody which can be detected, for example, by ELISA assay using free peptide adsorbed to a solid surface. The titer of anti-peptide antibodies in serum from an immunized animal may be increased by selection of anti-peptide antibodies, for instance, by adsorption to the peptide on a solid support and elution of the selected antibodies according to methods well known in the art.
[0212]Immunogenic epitope-bearing peptides of the invention, i.e., those parts of a protein that elicit an antibody response when the whole protein is the immunogen, are identified according to methods known in the art. For instance, Geysen et al., 1984, supra, discloses a procedure for rapid concurrent synthesis on solid supports of hundreds of peptides of sufficient purity to react in an enzyme-linked immunosorbent assay. Interaction of synthesized peptides with antibodies is then easily detected without removing them from the support. In this manner a peptide bearing an immunogenic epitope of a desired protein may be identified routinely by one of ordinary skill in the art. For instance, the immunologically important epitope in the coat protein of foot-and-mouth disease virus was located by Geysen et al. with a resolution of seven amino acids by synthesis of an overlapping set of all 208 possible hexapeptides covering the entire 213 amino acid sequence of the protein. Then, a complete replacement set of peptides in which all 20 amino acids were substituted in turn at every position within the epitope were synthesized, and the particular amino acids conferring specificity for the reaction with antibody were determined. Thus, peptide analogs of the epitope-bearing peptides of the invention can be made routinely by this method. U.S. Pat. No. 4,708,781 to Geysen (1987) further describes this method of identifying a peptide bearing an immunogenic epitope of a desired protein.
[0213]Further still, U.S. Pat. No. 5,194,392 to Geysen (1990) describes a general method of detecting or determining the sequence of monomers (amino acids or other compounds) which is a topological equivalent of the epitope (i.e., a "mimotope") which is complementary to a particular paratope (antigen binding site) of an antibody of interest. More generally, U.S. Pat. No. 4,433,092 to Geysen (1989) describes a method of detecting or determining a sequence of monomers which is a topographical equivalent of a ligand which is complementary to the ligand binding site of a particular receptor of interest. Similarly, U.S. Pat. No. 5,480,971 to Houghten, R. A. et al. (1996) on Peralkylated Oligopeptide Mixtures discloses linear C1-C7-alkyl peralkylated oligopeptides and sets and libraries of such peptides, as well as methods for using such oligopeptide sets and libraries for determining the sequence of a peralkylated oligopeptide that preferentially binds to an acceptor molecule of interest. Thus, non-peptide analogs of the epitope-bearing peptides of the invention also can be made routinely by these methods.
Removal or Reduction of Protease Activity
[0214]The present invention also relates to methods for producing a mutant cell of a parent cell, which comprises disrupting or deleting a nucleic acid sequence encoding the protease or a control sequence thereof, which results in the mutant cell producing less of the protease than the parent cell.
[0215]The construction of strains which have reduced protease activity may be conveniently accomplished by modification or inactivation of a nucleic acid sequence necessary for expression of the protease activity in the cell. The nucleic acid sequence to be modified or inactivated may be, for example, a nucleic acid sequence encoding the protease or a part thereof essential for exhibiting protease activity, or the nucleic acid sequence may have a regulatory function required for the expression of the protease from the coding sequence of the nucleic acid sequence. An example of such a regulatory or control sequence may be a promoter sequence or a functional part thereof, i.e., a part which is sufficient for affecting expression of the protease. Other control sequences for possible modification include, but are not limited to, a leader, a polyadenylation sequence, a propeptide sequence, a signal sequence, and a termination site.
[0216]Modification or inactivation of the nucleic acid sequence may be performed by subjecting the cell to mutagenesis and selecting for cells in which the protease producing capability has been reduced or eliminated. The mutagenesis, which may be specific or random, may be performed, for example, by use of a suitable physical or chemical mutagenizing agent, by use of a suitable oligonucleotide, or by subjecting the DNA sequence to PCR generated mutagenesis. Furthermore, the mutagenesis may be performed by use of any combination of these mutagenizing agents. Examples of a physical or chemical mutagenizing agent suitable for the present purpose include ultraviolet (UV) irradiation, hydroxylamine, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), O-methyl hydroxylamine, nitrous acid, ethyl methane sulphonate (EMS), sodium bisulphite, formic acid, and nucleotide analogues. When such agents are used, the mutagenesis is typically performed by incubating the cell to be mutagenized in the presence of the mutagenizing agent of choice under suitable conditions, and selecting for cells exhibiting reduced or no expression of protease activity.
[0217]Modification or inactivation of production of a protease of the present invention may be accomplished by introduction, substitution, or removal of one or more nucleotides in the nucleic acid sequence encoding the protease or a regulatory element required for the transcription or translation thereof. For example, nucleotides may be inserted or removed so as to result in the introduction of a stop codon, the removal of the start codon, or a change of the open reading frame. Such modification or inactivation may be accomplished by site-directed mutagenesis or PCR generated mutagenesis in accordance with methods known in the art.
[0218]Although, in principle, the modification may be performed in vivo, i.e., directly on the cell expressing the nucleic acid sequence to be modified, it is preferred that the modification be performed in vitro as exemplified below.
[0219]An example of a convenient way to inactivate or reduce production by a host cell of choice is based on techniques of gene replacement or gene interruption. For example, in the gene interruption method, a nucleic acid sequence corresponding to the endogenous gene or gene fragment of interest is mutagenized in vitro to produce a defective nucleic acid sequence which is then transformed into the host cell to produce a defective gene. By homologous recombination, the defective nucleic acid sequence replaces the endogenous gene or gene fragment. It may be desirable that the defective gene or gene fragment also encodes a marker which may be used for selection of transformants in which the gene encoding the protease has been modified or destroyed.
[0220]Alternatively, modification or inactivation of the nucleic acid sequence encoding a protease of the present invention may be performed by established anti-sense techniques using a nucleotide sequence complementary to the protease encoding sequence. More specifically, production of the protease by a cell may be reduced or eliminated by introducing a nucleotide sequence complementary to the nucleic acid sequence encoding the protease which may be transcribed in the cell and is capable of hybridizing to the protease mRNA produced in the cell. Under conditions allowing the complementary antisense nucleotide sequence to hybridize to the protease mRNA, the amount of protease translated is thus reduced or eliminated.
[0221]It is preferred that the cell to be modified in accordance with the methods of the present invention is of microbial origin, for example, a fungal strain which is suitable for the production of desired protein products, either homologous or heterologous to the cell.
[0222]The present invention further relates to a mutant cell of a parent cell which comprises a disruption or deletion of a nucleic acid sequence encoding the protease or a control sequence thereof, which results in the mutant cell producing less of the protease than the parent cell.
[0223]The protease-deficient mutant cells so created are particularly useful as host cells for the expression of homologous and/or heterologous polypeptides. Therefore, the present invention further relates to methods for producing a homologous or heterologous polypeptide comprising (a) culturing the mutant cell under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide. In the present context, the term "heterologous polypeptides" is defined herein as polypeptides which are not native to the host cell, a native protein in which modifications have been made to alter the native sequence, or a native protein whose expression is quantitatively altered as a result of a manipulation of the host cell by recombinant DNA techniques.
[0224]The methods of the present invention for producing an essentially protease-free product is of particular interest in the production of eukaryotic polypeptides, in particular fungal proteins such as enzymes. The protease-deficient cells may also be used to express heterologous proteins of interest for the food industry, or of pharmaceutical interest.
Use of Proteases in Industrial Processes
[0225]The invention also relates to the use of the protease according to the invention in a selected number of industrial and pharmaceutical processes. Despite the long term experience obtained with these processes, the protease according to the invention features a number of significant advantages over the enzymes currently used. Depending on the specific application, these advantages can include aspects like lower production costs, higher specificity towards the substrate, less antigenic, less undesirable side activities, higher yields when produced in a suitable microorganism, more suitable pH and temperature ranges, better tastes of the final product as well as food grade and kosher aspects.
[0226]In large scale industrial applications aimed at food or feed production, proteolytic enzymes are commonly used to improve aspects like protein solubility, extraction yields, viscosity or taste, texture, nutritional value, minimalisation of antigenicity or antinutritional factors, colour or functionality as well as processing aspects like filterability of the proteinaceous raw material. In these applications the proteinaceous raw material can be of animal or vegetable origin and examples include vegetable proteins such as soy protein, wheat gluten, rape seed protein, pea protein, alfalfa protein, sunflower protein, fabaceous bean protein, cotton or sesame seed protein, maize protein, barley protein, sorghum protein, potato protein, rice protein, coffee proteins, and animal derived protein such as milk protein (e.g. casein, whey protein), egg white, fish protein, meat protein including gelatin, collagen, blood protein (e.g. haemoglobin), hair, feathers and fish meal.
[0227]An important aspect of the proteases according to the invention is that they cover a whole range of pH and temperature optima which are ideally suited for a variety of applications. For example many large scale processes benefit from relatively high processing temperatures of 50 degrees C. or higher to control the risks of microbial infections. Several proteases according to the invention comply with this demand but at the same time exhibit no extreme heat stabilities so that they resist attempts to inactivate the enzyme by an additional heat treatment. The latter feature allows production routes that yield final products free of residual proteolytic activity. Similarly many feed and food products have slightly acidic pH values so that for their processing proteases with acidic or near neutral pH optima are preferred. A protease according to the invention complies with this requirement as well.
[0228]The specificity of endoproteases is usually defined in terms of preferential cleavages of bonds between the carboxyl of the amino acid residue in position P1 and the amino group of the residue in position P1' respectively. The preference may be conditioned predominantly either by P1 (e.g. positively charged residues in substrates for trypsin), by P1' (e.g. hydrophobic residues in cleavages by thermolysin) or by both P1 and P2 (e.g. specific cleavages between two positively charged residues by adrenal medulla serine endoprotease). In some cases more distant residues may determine the cleavage preference, e.g. P2 for streptococcal peptidase A. Some residues are known to influence cleavages negatively; it is well known that bonds with proline in position P1' are resistant to the action of many proteases. Most endoproteases cleave preferentially either in a hydrophobic environment or in the proximity of negatively charged residues. For example, industrially available endoproteases like chymotrypsin (obtained from bovine pancreas) or subtilisin, neutral metallo endoprotease or thermolysin (all obtained from Bacillus species) tend to favour cleavage "behind" hydrophobic amino acids like -Phe, -Leu and -Tyr. Other industrially available endoproteases are trypsin (obtained from bovine pancreas) preferring cleavage behind -Arg and -Lys and papain (a complex mixture of various enzymes including proteases obtained from papaya fruits) preferring cleavage behind -Arg.
[0229]In contrast, peptide bonds formed by small sized residues such as Ala, Gly, Ser, Thre as well as Ile and Pro are poor substrates (Keil, B et al.; Protein Seq Data Anal (1993) 5; 401-407). This situation has a profound implications for the pharmaceutical, the food and beverages, the agro and even the chemical industry. A protease according to the invention exhibits uncommon cleavage preferences.
[0230]The exopeptidases act only near the ends of polypeptide chains. Those acting at a free N-terminus liberate a single amino acid residue (socalled aminopeptidases) or a dipeptide or a tripeptide (socalled dipeptidyl-peptidases and tripeptidyl-peptidases) Those acting at a free C-terminus liberate a single residue (socalled carboxypeptidases) or a dipeptide (socalled peptidyl-dipeptidases) The carboxypeptidases are allocated to three groups on the basis of catalytic mechanism i.e. serine-type carboxypeptidases, metallocarboxypeptidases and cystein-type carboxypeptidases. Other exopeptidases are specific for dipeptides (socalled dipeptidases) or are able to cleave peptide linkages other than those of alpha-carboxyl or alpha-amino groups (socalled omega peptidases). Examples of such new omega peptidases are the pyroglutamyl-peptidase and the acylaminoacyl-peptidase as identified in the present invention (see Table 1, genes 18 and 45 respectively).
[0231]Typical examples of industrial application which depend on the use of pure endoproteases and in which the protease according to the invention can be expected to deliver a superior performance include the processing of materials of vegetable or animal origin. These processing steps can be aimed at modifying a large array of characteristics of either the crude material or the (partially) purified protein fraction. For example, these processing steps can be aimed at maximising product solubilities, filterabilities, separabilities, protein extraction yields and digestibilities or minimising toxicities, off-tastes and viscosities. Furthermore the treatment can be directed at altering physico-chemical characteristics of the crude material or the purified (or partially purified) protein. These advantages apply not only if the endoprotease according to the invention is applied as a processing aid in industrial applications but also if applied as an active enzyme component in animal feed. Specifically the endoprotease according to the invention can be applied as bread improver in the bakery industry, e.g. to retard the staling of bread or to diminishing the viscosity of doughs. Or the endoprotease can be used in the beer and wine industry to prevent or to minimise the formation of undesirable protein hazes. Alternatively it can be used in the beer industry to optimise the protein extraction yields of cereals used in the preparation of the wort. Furthermore, it can also be advantageously used in the dairy industry as a milk clotting agent with superior characteristics or to optimise the texturising, foaming or setting characteristics of various milk components. Another application in the dairy industry is the use of the new protease in the preparation of Enzyme Modified Cheeses (EMC's).
[0232]Moreover, various proteinaceous substrates can be subjected to an endoprotease according to the invention, usually in combination with other proteolytic enzymes to obtain hydrolysates for medical or non-medical applications. Here the endoprotease according to the invention is surprisingly effective in achieving a complete hydrolysis of the proteinaceous substrate so that even protease resistant parts are fully hydrolysed, the endoprotease is also surprisingly active in minimising the allergenicity of the final hydrolysate or in suppressing the formation of bitter off-tastes.
[0233]More specifically the endoprotease according to the invention is characterised by its preference for cleaving proteins at unusual peptide bonds, especially with the small size amino acid residues of Ala, Gly, Ser and Thr, or the residues Ile and Pro in either the P1 or the P1' position (Keil, B et al.; Protein Seq Data Anal (1993) 5; 401-407). As the result those fractions of the proteinaceous starting materials that resist hydrolysis upon using prior art endoproteases, can be dissolved and hydrolysed using the endoprotease according to the invention. Non limiting examples of such protease resistant fractions include socalled extensins in plant materials and collagen, gelatin but also specific milk components in material of animal origen.
[0234]Various feedstuffs such as e.g. soybeans contain trypsin inhibitors. These proteins inhibit trypsin activity in the GI-tract of e.g. pigs and poultry. This trypsin inhibiting activity results in sub-optimal protein digestibility in these animals resulting in increased waste production and poor economics. This problem may partly be overcome by toasting soybeans at high temperatures. Two different types of trypsin inhibitors have been identified in soybeans, i.e. the Bowman-Birk type trypsin inhibitors and the Kunitz type trypsin inhibitors.
[0235]This invention now provides an alternative way to degrade trypsin inhibiting activity over toasting, in that it provides a cysteine proteases (EC 3.4.22, table 1) capable of cleaving at Leucine176-Aspartate177 peptide bond near the carboxyl-terminus of the Kunitz type trypsin inhibitor (as reviewed by Wilson (1988) in CRC Critical Reviews in Biotechnology 8 (3): 197-216). This results in inactivation of this trypsin inhibitor in soybean. It was surprisingly found that the cysteine proteases secreted by the fungus Aspergillus niger fulfilled these criteria far better than similar enzymes derived from other organisms.
[0236]Proteases are also widely used in the art of cheese-making. In the production of cheese it is necessary to coagulate the cheese milk to be able to separate the cheese matters e.g. casein from the whey. Several milk coagulating enzymes, also referred to as coagulants, have been described and include (bovine) chymosin, bovine pepsin, porcine pepsin as well as microbial enzymes like Rhizomucor miehei protease, Rhizomucor pusillus protease and Cryptonectria parasitica protease. Chymosin can be obtained from calf stomachs but can also be produced microbially by for example Kluyveromyces lactis. All these enzymes are characterized by having specificity for the peptide bond between residue 105 (phenylalanine) and residue 106 (methionine) or the bond adjacent to that in K-casein. This means that by employing these enzymes in cheese making, the K-casein is split at the junction between para-K-casein and the macro-peptide moiety called glycomacropeptide (GMP) carrying the negative charges. When this occurs the macropeptide diffuses into the whey, its stabilizing effect on the solubility of the casein micelles is lost, and the casein micelles can start to aggregate once sufficient kappa-casein has been hydrolyzed. For further elaboration on the enzymatic coagulation of milk (e.g. D. G. Dalaleish in Advanced Dairy Chemistry vol. 1 ed by P. F. Fox, Elsevier, London, 1992.
[0237]The currently available coagulants allow for a rather high yield of cheese, however, it should be realised that due to the enormous volumes of cheese produced, an increased yield in the order of magnitude of tenths of percent points may constitute a substantial economical advantage. Consequently there is a great need in the art for coagulants with an (even slightly) improved yield.
[0238]Coagulants are characterized by their high substrate specificity, which is, however, dependent on pH and temperature. In a typical cheese making process the pH will change from the initial pH 6.3 to lower pH values in the range of 4.5-5.5, the end-value depends on the conditions used during the cheese production process. Some coagulants are more sensitive to pH changes than others. The Rhizomucor pusillus protease for example is more sensitive to pH changes than chymosin. Besides pH, also other parameters like temperature and water content may affect the protease specificity. It is well known that most coagulants show a changing substrate specificity with changing pH, resulting in altered proteolytic activity in later stages of the cheese making process. It is also well known that coagulants differ in the extent of casein proteolysis; they may also show differences in the peptide patterns produced during proteolysis. These are relevant factors during cheese ripening and may affect cheese properties like taste, flavor and texture. In some cases coagulants give rise to undesired effects like the formation of bitter tasting peptides or off-taste. In addition, changes in proteolytic specificity may lead to a reduction in yield. Pepsin, a well known component in many bovine chymosin preparations, is an example of a protease that gives rise to lower yields and taste effects as compared to pure chymosin. There is still a need for coagulants with give rise to new, improved cheese texture and taste. Such new coagulants result in the accelerated development of taste and texture profiles related to cheese aging, therewith providing a substantial economical benefit.
[0239]It is well known that free amino acids are very important in taste and flavour generation. Especially the amino acids leucine, phenylalanine, methionine and valine play an important role in the generation of typical cheese taste and flavor components. The free amino acids are converted via fermentation by micro organisms that are added during the cheese manufacturing process into the actual flavor and taste generating compounds like methanediol, dimethyldisulphide, methylpropanoic acid and methylpropanal. Exo-peptidases play an important role in the generation of free amino acids. They can only be effective, however, when they are combined with an endo-protease of appropriate specificity. Appropriate combinations of exo- and endo-peptidases can be used in cheese making, resulting in the manufacture of cheeses with new and improved taste profiles.
[0240]The enzymes according to the invention may be used to hydrolyze proteinaceous materials of animal origin such as whole milk, skim milk, casein, whey protein or mixtures of casein and whey protein. Such mixtures of casein and whey protein may be used, for example, in ratios similar to those found in human milk. Furthermore, the enzyme mixture according to the invention may be used to hydrolyze proteinaceous materials of plant origin such as, for example, wheat gluten malted or unmalted barley or other cereals used for making beer, soy milk, concentrates or isolates thereof, maize protein concentrates and isolates thereof, and rice proteins.
[0241]Within the area of large scale industrial processes, some applications rely on the use of endoproteases only whereas in other applications combinations of endoproteases with exoproteases are essential. Typical examples which depend on the use of pure endoproteases and in which the protease according to the invention can deliver a superior performance include applications like the processing of soy or peas or cereals proteins aimed at minimising viscosities or optimising foaming or other physics-chemical characteristics, bread improvers in the bakery industry also aimed at diminishing the viscosity of doughs, processing aids in the beer and wine industry aimed at the prevention of protein hazes or optimising the extraction yields of cereals, feed additives in the bio industry aimed at enhancing intestinal absorption or modulating microbial activities in the gut, processing aids in the dairy industry aimed at optimising the clotting, foaming or setting characteristics of various milk components. Moreover, v For specific market segments proteins derived from milk or soy or collagen are exposed to proteases to produce socalled protein hydrolysates. Although the main outlets for these protein hydrolysates are infant formula and food products for hospitalised persons, products intended for persons with non-medical needs, such as athletes or people on a slimming diet form a rapidly growing segment. In all of these applications protein hydrolysates offer attractive advantages such as lowered allergenicities, facilitated, gastro-intestinal uptake, less chemical deterioration of desirable amino acids like glutamine and cystein and finally, absence of proteinaceous precipitations in acid beverages during prolonged storage periods. All these advantages can be combined if the hydrolysate is offered as a mixture of di- and tripeptides. However, currently all commercially available hydrolysates are produced by combining several endoproteases. The latter approach implies a non-uniform and incomplete degradation of the protein. To obtain the desired mixture of di- and tripeptides, a hydrolysis process involving a combination of various di- and tripeptidylpeptidases would be ideal. Unfortunately, only few of these enzymes from food grade and industrially acceptable microorganisms are known, let alone industrially available. According to the invention several of highly useful di- and tripeptidylpeptidases are economically obtainable in a relatively pure state. Preferred are those di- or tripeptidylpeptidases that exhibit a low selectivity towards the substrate to be cleaved, i.e. exhibit minimal amino acid residue cleavage preferences only. Preferred are combinations of those di- or tripeptidylpeptidases that hydrolyse high percentage of the naturally occurring peptide bonds. Despite this high activity to naturally occurring peptide bonds, a total hydrolysis to free amino acids is prevented by the nature of the di- and tripeptidylpeptidases. Also preferred are those di- or tripeptidylpeptidases that are optimally active between pH 4 to B and exhibit adequate temperature stability. Adequate temperature stability implies that at least 40%, preferably at least 60%, more preferably between 70 and 100% of the initial hydrolytic activity survives after heating the enzyme together with the substrate for 1 hour at 50 degrees C.
[0242]Although the process towards an efficient production of mixtures di- or tripeptides or di- and tripeptides hinges on the availability of the enzymes according to the invention, the first enzyme incubation with the proteinaceous substrate will usually be an endoprotease. Preferably an endoprotease with a broad spectrum endopeptidase suited for the situation, e.g. subtilisin (Delvolase from DSM), neutral metallo protease (Neutrase from NOVO) or thermolysin (Thermoase from Daiwa Kasei) for the near neutral conditions and pepsin or aspergillopepsin (e.g. Sumizyme A P from Shin Nihon, Japan) for the acidic conditions. Aim of this first digestion is to improve the solubility, to reduce the viscosity and to reduce the heat setting characteristics of the water/protein mixture. Furthermore this pretreatment with an endonuclease is essential to create enough starting points for the di- and tripeptidylpeptidases hereby accelerating the process of di- or tripeptide formation. Optionally a protease intended for debittering of the hydrolysate can be included in this stage of the process or later, together with the di- or tripeptidylpeptidases.
[0243]Main aim of the latter hydrolysates is to minimize the allergenicity of the product or to facilitate gastrointestinal uptake. In the production of such hydrolysates the use of dipeptidyl- and tripeptidyl-peptidases is of special importance as hese s offer an efficient way for producing hydrolysates.
[0244]Other applications in these food and feed industries totally rely upon combinations of one or more endoprotease(s) with one or more exoprotease(s). Such combinations of an endoprotease with an exoprotease are typically used in industries to improve aspects like taste and colour of the final product. The reason for this is that the development of taste and colour is largely dependent upon the presence of free amino acids. Free amino acids can not only be obtained by exoproteases such as carboxypeptidases and aminopeptidases but also by peptidyl-dipeptidases. If combined with endoproteases or even dipeptidyl- or tripeptidyl-peptidases, carboxypeptidases, aminopeptidases and peptidyl-dipeptidases can create larger quantities of free amino acids in less time. However, in all of these processes an uncontrolled release of amino acids or even non-proteinaceous components should be avoided to minimise undesirable side reactions.
[0245]Though free amino acids as such, can elicit a number of taste impressions, these taste impressions are very basic (bitter, sweet, sour and "umami") and the amino acid concentration required for perceiving these tastes are high. Despite these high threshold values, free amino acids are able to create major sensory effects at much lower concentration ranges through a number of flavour enhancing mechanisms. One of these mechanism involves the combination of free amino acids with sugars in so-called Maillard reactions. Compared with free amino acids, with these Maillard products overwhelmingly complex flavour and odour systems can develop with threshold values that are several orders of magnitude lower than those recorded for the free amino acids. Maillard products are formed at elevated temperatures usually during cooking, baking or roasting when preparing food or feed products. During these treatments both colour and a large array of aromas develop. In these reactions amino groups react with reducing compounds as a first step and ultimately leading to a whole family of reaction pathways. In foods or feeds the amino compounds involved are predominantly free amino acids which are released from the proteinaceous raw material by various proteases and the required reducing compounds primarily represent reducing sugars. The implication is that during the processsing of the raw material undesired release of free amino acids and sugars should be avoided to minimise off tastes that could be generated during subsequent heating steps as e.g. during spray drying or sterilisation. The latter notion emphasises once more the benefits of superior purity and low in-use costs of the enzyme according to the invention.
[0246]Apart from Maillard reactions, amino acids can also undergo important chemical transitions at ambient temperatures. The latter type of transitions are enzyme dependent and are quite common in fermented foods such as beer, yoghurt, cheese ripening and meat and wine maturation processes. In these fermentation processes, free amino acids are liberated from the raw materials used by the proteases added or by proteolytic enzyme activity from the raw material or the microbial starters used. During the maturation phase microbial metabolic activity then converts the free amino acids into derivatives with increased sensoric properties. For example, L-leucine, L-isoleucine and L-valine lead to the formation of valuable fusel alcohols like amylalcohols and isobutanol in beer fermentation. Similarly cheese volatiles such as methanethiol and dimethyldisulphide have been traced back to the occurrence of methionine in cheese as well as methylpropanoic acid and methylpropanal to valine. Finally the free amino acid glutamate and can create strong savoury enhancing effects because of its synergy with the breakdown products of RNA, so-called 5'-ribonucleotides. If combined with proper concentrations of 5'-ribonucleotides such as 5'-IMP and 5'-GMP, the detection threshold of the umami taste generated by glutamate is known to be lowered by almost two orders of magnitude.
[0247]In order to obtain pronounced and precise taste effects in all of these processes, the proteinaceous substrates should be hydrolysed using a combination of an endo- and an exoprotease, wherein at least one of the endo or exoprotease, preferably both the endo- and exoprotease, are pure and preferably selective towards a specific set of amino acid(s) or preferentially release the preferred amino acid(s). So preferred proteases are characterised by a high selectivity towards the amino acid sequences that can be cleaved which notion makes the enzyme category in Aspergillus known as "maturases" of particular importance.
[0248]Apart from the food and feed industries, proteases are also commonly applied by the chemical, pharmaceutical, diagnostic and personal care industries.
[0249]In the personal care industry proteases are used to create peptides which are added to a variety of products to improve aspects like skin feel, gloss or protection. Moreover there is a new tendency towards direct topical application of the protease. Very similar to the enzyme use in the leather industry, the prime aim in the latter application is to clean, dehair and soften the skin.
[0250]In the chemical and pharmaceutical industry proteases are being developed as valuable tools in producing costly ingredients or intermediates. In these industries proteases are not only used because of their hydrolytic capacity but also because of their capacity to synthesise peptides from natural or non-natural amino acids. The latter option is clearly demonstrated by the possibility to synthesize aspartame from its amino acid based building blocks by using an endoprotease like thermolysin.
[0251]Unlike the situation in the food and feed industry, the stereo- and regioselectivity of proteases are also considered important assets although unusual reaction conditions may be needed to accomplish the desired chemical transformation. Typical examples of the application of proteases in this industry include the use of endoproteases, aminopeptidases as well as carboxypeptidases in the production of various intermediates for drugs like insulin, antibiotics, renin and ACE-inhibitors An overview of such uses is presented in Industrial Biotransformations, A. Liese, K. Seelbach, C. Wandrey, Wiley-VCH; ISBN 3-527-30094-5.
[0252]In view of the desired specificities, stereo- and regioselectivities, the absence of side activities and resistance to unusual reaction conditions such as high solvent concentrations, the improved performance of the protease according to the invention offers substantial advantages.
[0253]From a pharmaceutical point of view the role of proteases is illustrated by a substantial number of references in Martindale's, "The Extra Pharmacopoeia" (Pharmaceutical Press, London, UK). Moreover the important role of very specific proteases in regulating all kinds of biological processes is illustrated by the fact that many hormones become active only after the processing of an, mostly inactive, precursor molecule by such a very specific protease. Inhibitors active towards certain categories of such specific proteases have been implicated in the development of all kinds of new drugs. Therefore new and effective inhibitors for protease may now be identified using the sequences provided herein.
[0254]The entire disclosure of each document cited herein is hereby incorporated by reference
TABLE-US-00001 TABLE 1 SEQ ID number Gene cDNA Protein Function of encoded protein EC number 1 58 115 Pepsin A3 EC3.4.23.1 2 59 116 Metalloprotease EC3.4.24.56 3 60 117 acylaminoacyl-peptidase EC3.4.19.1 4 61 118 Tripeptidylaminopeptidase EC3.4.14.- 5 62 119 serine carboxypeptidase EC3.4.16.6 6 63 120 Serine endoprotease EC3.4.21.- 7 64 121 Carboxypeptidase Y EC3.4.16.5 8 65 122 aspergillopepsin II - hom EC3.4.23.19 9 66 123 Tripeptidyl peptidase EC3.4.14.9 10 67 124 Tripeptidyl peptidase EC3.4.14.9 11 68 125 aspergillopepsin II - hom EC3.4.23.19 12 69 126 Tripeptidyl peptidase EC3.4.14.9 13 70 127 Metalloprotease EC3.4.24.- 14 71 128 aspergillopepsin I EC3.4.23.18 15 72 129 Pepsinogen E EC3.4.23.25 16 73 130 aspergillopepsin I - hom EC3.4.23.18 17 74 131 aspergillopepsin II EC3.4.23.19 18 75 132 Pyro-Glu peptidase EC3.4.19.3 19 76 133 dipeptidyl peptidase EC3.4.14.2 20 77 134 Secr. aminopeptidase EC3.4.11.10 21 78 135 alkaline D-peptidase EC3.4.16.4 22 79 136 Carboxypeptidase EC3.4.16.1 23 80 137 Carboxypeptidase EC3.4.16.1 24 81 138 Carboxypeptidase-II EC3.4.16.1 25 82 139 aspartic proteinase EC3.4.23.- 26 83 140 Tripeptidyl peptidase EC3.4.14.9 27 84 141 Carboxypeptidase EC3.4.16.1 28 85 142 cysteine proteinase EC3.4.22.- 29 86 143 Metallocarboxypeptidase EC3.4.17.- 30 87 144 Subtilisin hom. EC3.4.21.62 31 88 145 Carboxypeptidase Y EC3.4.16.5 32 89 146 Metalloprotease EC3.4.24.- 33 90 147 Carboxypeptidase Y EC3.4.16.5 34 91 148 Metalloprotease EC3.4.24.- 35 92 149 Tripeptidyl peptidase EC3.4.14.9 36 93 150 Aspartic protease EC3.4.23.24 37 94 151 Aspartic protease EC3.4.23.24 38 95 152 Pepsin A3 EC3.4.23.1 39 96 153 Aspartic protease EC3.4.23.24 40 97 154 Aspartic protease EC3.4.23.24 41 98 155 Kex EC3.4.21.61 42 99 156 Serine protease EC3.4.21.- 43 100 157 Glutamyl endoprotease EC3.4.21.82 44 101 158 aspergillopepsin II - hom EC3.4.23.19 45 102 159 acylaminoacyl-peptidase EC3.4.19.1 46 103 160 Tripeptidylaminopeptidase EC3.4.14.- 47 104 161 serine carboxypeptidase EC3.4.16.6 48 105 162 Gly-X carboxypeptidase EC3.4.17.4 49 106 163 aspartic proteinase EC3.4.23.- 50 107 164 Tripeptidyl peptidase EC3.4.14.9 51 108 165 Carboxypeptidase-I EC3.4.16.1 52 109 166 serine carboxypeptidase EC3.4.16.6 53 110 167 serine carboxypeptidase EC3.4.16.6 54 111 168 Secr. aminopeptidase EC3.4.11.10 55 112 169 Prolyl endopeptidase EC3.4.21.26 56 113 170 aspergillopepsin I - hom EC3.4.23.18 57 114 171 Aminopeptidase EC 3.4.11.-
EXAMPLES
Example 1
Assaying Proteolytic Activity and Specificity
[0255]Protease specificity may be explored by using various peptide substrates. Synthetic substrates are widely used to detect proteolytic enzymes in screening, in fermentation, during isolation, to assay enzyme activity, to determine enzyme concentrations, to investigate specificity and to explore interaction with inhibitors. Peptide p-nitroanilides are preferably used to assay protease activity as the activity can be followed continuously and therefore allow for kinetic measurement. The cleavage of peptide p-nitroanilides can be followed by measuring the increase in adsorption at 410 nm upon release of the 4-nitroanilide. Paranitroanilide substrates are generally used for serine and cysteine proteases. In addition peptide thioesters and 7-amino-p-methylcoumarin peptide derivates are used. Peptide thioesters are very sensitive substrates for serine and metalloproteases that exhibit relatively high turnover rate since the thioester bond is easier to cleave than the amide bond. Cleavage of thiolesters may be followed with a thiol reagent such 4,4-dithiopyridine (324 nm) or 5,5-dithiobis 2-nitrobenzoic acid (405 nm). The same increased turnover rate is usually observed for the cleavage of ester bonds relative to amide bond. The most well known substrates to assay the esterase activity of proteases are p-nitrophenol derivates. The release of p-nitrophenol can be monitored at different wavelength dependent on the pH that is used, eg around neutral pH a wavelength of 340 nm is used while above pH 9 monitoring is done around 405 nm. In addition the hydrolysis of esters can also be followed by titration using pH-stat equipment. In case of qualitative measurement of esterase activity pH sensitive dyes can be applied.
[0256]As an alternative, peptides may be attached to a fluorescent leaving group. Proteolysis is accompanied by an increase in fluorescence when monitored at the appropriate wavelengths. Peptidyl 2-naphtylamides and peptidyl 4-methyl-7-coumarylamides are commonly used. The release of for example 7-amino-4 methylcoumarin is measured using an excitation wavelength of 350 nm and an emission wavelength of 460 nm. The use of 7-amino-4 trifluoromethylcoumarin has the advantage of the leaving group being both chromogenic (absorbtion 380 nm) as well as fluorogenic (excitation 400 nm, emission 505 nm). When it is essential that at both sides of the scissile bond an amino acid is present, the introduction of a group that quenches the fluorescence might be useful. The general characteristics of such substrates is that the peptide sequence separates a fluorescent donor group from an acceptor group that acts as a quencher of fluorescence. Cleavage of a peptide bond between the quenching group and the fluorophore will lead to substantial increase in fluorescence. Several donor-acceptor pairs have been reported, including o-aminobenzoic acid (Abz) as the donor and 2, 4 dinitrophenyl (Dnp) as the acceptor, 5-[(2'aminoethyl)-amino]naphtalenesulfonic acid (EDANS) as the donor and 4-[[4'-(dimethylamino)phenyl]azo]-benzoic acid (DABCYL) as the acceptor. The Abz/EDDnp represents a very convenient donor-aceptor pair since after total hydrolysis, the fluorescence increases by a factor 7 to 100 and the absorption spectrum of EDDnp does not change with pH. Moreover, the peptide sequence may contain up to 10 residues without loss of the quenching effect. As the size of the connecting peptides increases, the position of the scissile bond may become less specific. Therefore in addition to establishing whether proteolysis occurred, additional analysis of the products may be required. This may be done by analysing and separating the produced peptides by HPLC and determining the amino acid sequence of the fragments. In addition the peptide composition of the digest may be directly analysed by using combined HPLC/mass-spectroscopy technique.
[0257]Apart from using peptides of a defined sequence also synthetic peptide libraries can be used to study protease specificity. Peptides are synthesised by solid phase synthesis in random or semi-random fashion. E.g. Meldal et al. (PNAS USA 91, 3314, 1994) report the preparation of a family of protease substrates by starting with H-Lys(Abz)-resin, extending the resin with peptides to a length of six aminoacids, and finally coupling Tyr(NO2) to the peptides. Each resin bead has a unique sequence and on treatment with the proteases the most susceptible becomes fluorescent as the Tyr(NO2) containing peptide is released. Sequence analysis of the peptides on the susceptible will give information on the specificity of the protease.
[0258]Protease activity is usually expressed in units. Generally the international standard unit (IU) is defined as the amount of enzyme, which under defined conditions transfers one micromole of substrate per minute. Specifically with proteases the IU would relate to the hydrolysis of one micromole peptide bond per minute. However in the case of protease units deviations of the international definition are more rule than exception. Where with the model peptides, which are cleaved specifically at one bond the calculation of IU's is strait-forward, for proteinacious substrates where the protease can cleave at various positions to a various degree many deviating unit definition are used. Apart from a definition of the unit used, any hydrolysis experiment requires an adequate description of the conditions under which the units are measured. Such conditions comprise e.g. the substrate concentration, the enzyme-substrate ratio, the pH and temperature. Typical assays for determining the specific activity of a proteases comprise a proteinacious substrate such as for example denaturated hemoglobin, insulin or casein. The polypeptide substrate is digested by a protease at fixed conditions during a fixed time interval. Undigested and large polypeptides are precipitated with TCA and TCA soluble product is determined by measuring absorbance at 220 or 280 nm, or by titrating the soluble peptides with folin reagent, ninhydrin, fluoro 2,4, dinitrobenzene/dansylchloride, TNBS method or fluorescein. Instead of labeling the product after hydrolysis, also polypeptide substrates may be used which are already labeled by specific dyes or fluorophores such as for example fluorescein. In addition standard methods of amino acid analysis may be applied using standard laboratory analyzers. In order to hget insight in the size distribution of the peptides generated by a protease, gel chromatography experiments may be performed. In addition to this HPLC using reverse phase techniques is applied in order to get better resolution of the peptide patterns generated by the protease.
[0259]The course of the hydrolysis of proteinacious substrates is usually expressed in the degree of hydrolysis or DH. In case pH-stat is used to follow the course of hydrolysis, DH can be derived from the base consumption during hydrolysis (Enzymatic Hydrolysis of Food Protein, J. Adler-Nissen, 1986, Elsevier Applied Science Publishers LTD). The DH is related to various useful functional properties of the hydrolysate such as solubility, emulsifying capacity, foaming and foam stability, whipping expansion, organoleptic quality. In addition taste is an important aspect of food grade hydrolysates. Bitterness can be a major problem in protein hydrolysates. Termination of the hydrolysis reaction may be done by changing the pH, heat inactivation, denaruring agents such as SDS, acetonitril etc.
[0260]Polypeptides shown in Table 1 were expressed and at least partially purified according to standard procedures known in the art. They were analysed according to at least one of the methods described above and found to have the activities listed in Table 1.
Example 2
Direct Determination of the kcat/Km Ratio for Protease Substrates
[0261]Synthetic substrates can be used to monitor the enzymatic activity during purification, to determine enzyme concentration, to determine inhibition constants or to investigate the substrate specificity. Determination of the kcat/Km ratio gives a measurement of the substrate specificity. It allows to compare the specificity of different substrates for a same enzyme or the comparison of hydrolysis rates with different enzymes cleaving the same substrate. This ratio has a unit of a second order rate constant and is then expressed as 1/(concentrationtime). Substrates having a kcat/Km ratio in the range 10.5-10.6 M-1sec-1 are considered to be very good substrates i.e good affinity and rapid turn-over. However, some substrates may be very specific with kcat/Km values in the 10.4 M-1sec-1 range.
[0262]The kcat/Km ratio may be calculated after determination of individual parameters. In that case, Km and Vm may be obtained from various linear plots (e.g Hanes or Cornish-Bowden method) or by a non-linear regression method. Knowing that Vm=kcatEt (where Et is the final active enzyme concentration then kcat=Vm/Et. Determination of the kcat/Km ratio by the previous method may be prevented when product or substrate inhibition occur, or when substrate precipitates at high concentration. It is however possible to obtain an accurate value of the kcat/Km ratio working under first-order conditions i.e at a substrate concentration far below the estimated Km. In these conditions, the Michaelis-Menten equation: v=(VmS)/(Km+S) becomes:
v=(VmS)/Km since S<<Km
or v=(Vm/Km)S=kobsS=-dS/dt
which integrates as InS=-kobst+InSo where So is the starting substrate concentration and S the substrate concentration at a given time. The velocity is proportional to the substrate concentration. In other words, the substrate hydrolysis obeys a first order process with kobs as the first-order rate constant. kobs=Vm/Km=(kcatEt)/Km since Vm=kcatEt
[0263]A continuously recording of the substrate hydrolysis will allow the graphical determination of kobs from the InS vs time graph. The kcat/Km ratio is simply inferred from kobs providing the active enzyme concentration is known:
kcat/Km=kobs/Et
[0264]Assay method: Use a starting substrate concentration far below the estimated Km and a low enzyme concentration to allow the substrate hydrolysis to be recorded. You will obtain a first-order curve for the product generation:
[0265]After total hydrolysis of the substrate, the absorbance (or fluorescence units) of the product will allow the accurate determination of So, since Pt=Sokobs is determined from the slope of the InS vs time graph or alternatively using a fitting software (Enzfitter, SigmaPlot . . . ).
[0266]NB: Do not forget to calculate the substrate concentration for any given time from the product concentration (S=So-P) since plotting P vs time would not provide the correct kobs (dP/dt=kobsS does not integrate in the same way).
[0267]Alternatively, one can measure successive t1/2 (half-time) from the product apparition curve since in a first order process:
t1/2=ln 2/lkobs=0.693/kobs then kobs=0.693/t1/2
[0268]Using this method allows to check that you have a true first order decay (identical values for the successive t1/2).
Example 3
Inactivating Protease Genes in Aspergillus
[0269]The most convenient way of inactivating protease genes in the genome of Aspergillus is the technique of gene replacement (also called "one step gene disruption"). The basics of this technique have been described by Rothstein R J in Meth. Enzymol. 101, p 202, 1983. Essentially the technique is based on homologous recombination of transformed DNA fragments with the genomic DNA of a fungal cell. Via double crossover the gene to be inactivated is (partly) replaced by the DNA fragment with which the cell is transformed. Preferably the transformed DNA fragment contains a selectable marker gene for Aspergillus niger. Basically the manipulation of DNA and generation of a inactivation construct are done using general molecular biological techniques. First, genomic DNA is isolated from the Aspergillus niger strain that is later on used for the inactivation of the protease gene. Genomic DNA of A. niger can be isolated by any of the techniques described, e.g. by the method described by de Graaff et al. (1988) Curr. Genet. 13, 315-321, and known to the person skilled in the art. This genomic DNA is used as template for amplification of the flanking regions of the protease gene by using the polymerase chain reaction (PCR; Sambrook et al. (1989) Molecular cloning, a laboratory manual, 2nd edition, Cold Spring Harbor Laboratory Press, New York). With flanking regions is meant here the non-coding regions upstream and downstream of the protease gene that will be inactivated. Preferably the flanking regions should each be more than 1.0 kb in length.
[0270]Two single stranded DNA oligonucleotides are used for the priming of the PCR amplification of each flanking region. For the 5'-flanking region, one primer is homologous to a DNA sequence upstream of the start of the coding sequence of the protease gene. Preferably the homologous region is located more than 1.0 kb upstream of the translation start site. The second primer is homologous to the complementary and inverse DNA sequence located immediately upstream of the coding sequence of the protease gene.
[0271]For the 3'-flanking region, one primer is homologous to the DNA sequence immediately downstream of the coding sequence of the protease gene. The second primer is homologous to a complementary and inverse DNA sequence located preferably more than 1.0 kb downstream of the coding sequence of the protease gene.
[0272]The DNA sequence included in all primers and homologous to the A. niger genome should be minimally 15 nucleotides in length, preferably more than 18 nucleotides in length. Most conveniently, all primers should contain a DNA sequence coding for the recognition site of suitable restriction enzymes upstream of the sequence that is homologous to the A. niger genome. These extra recognition sites facilitate the cloning process.
[0273]Both primers and the genomic DNA of A. niger are used in a PCR reaction under conditions known to those skilled in the art. The annealing temperature of the primers can be calculated from the part of the DNA sequence that is homologous to the A. niger genome. Both fragments containing the 5'-flanking region and the 3'-flanking region are cloned into a vector that can be propagated in E. coli using general molecular biological techniques. A gene that can be used as selection marker in Aspergillus niger is then cloned in between the two flanking regions. Most conveniently the marker gene is under control of a promoter that comes to expression in A. niger, preferably an endogenous A. niger promoter. The orientation of the insertion of the marker gene is preferably in the same direction as the original protease gene. The final inactivation fragment contains the 5'-flanking region, a selection marker gene preferably under control of a A. niger endogenous promoter, and the 3'-flanking region, all in this direction and orientation. DNA of the final construct is cloned into a vector that can be propagated in E. coli.
[0274]The inactivation construct is digested with suitable restriction enzymes to remove the E. coli vector sequences and the inactivation fragment is isolated using standard techniques (Sambrook et al. (1989) Molecular cloning, a laboratory manual, 2nd edition, Cold Spring Harbor Laboratory Press, New York). Finally Aspergillus niger is transformed with the inactivation fragment using a method described in literature, e.g. by the method described by Kusters-van Someren et al. (1991) Curr. Genet. 20, 293-299. Transformed cells are selected by plating the transformation mixture on agar plates that are selective for growth of Aspergillus niger strains that do express the marker gene. After purification of the transformed Aspergillus strains by replica plating, a representative number of strains is analysed by Southern blotting using standard methods (Sambrook et al. (1989) Molecular cloning, a laboratory manual, 2nd edition, Cold Spring Harbor Laboratory Press, New York). Therefore, genomic DNA of mycelium of transformed strains is isolated and digested with suitable restriction enzymes. Restriction fragments are separated using agarose gel electrophoresis, blotted to nitrocellulose membranes and probed with a labeled fragment of the marker gene. Hybridization and washing is under stringent conditions. Strains that contain labeled restriction fragments of the correct length are considered correct.
[0275]Using this method A. niger strains can be selected with an inactivated protease gene of choice.
Example 4
Isolating Proteases by Ion Exchange Chromatography
[0276]Small quanties of the protease encoded by the nucleotide sequence as provided herein are obtained by constructing an expression plasmid containing the relevant DNA sequence, transforming an A. niger strain with this plasmid and growing the A. niger strain in a suitable medium. After collecting the broth free of contaminating cells, the protease sought can be purified.
[0277]To isolate the protease as encoded by the provided nucleotide sequence in an essentially pure form several strategies can be followed. All of these strategies have been adequately described in the relevant scientific literature (see for example the Protein Purification Handbook, 18-1132-29 Edition M as published by Amersham Pharmacia Biotech, Uppsala, Sweden). A procedure which is applicable to purify proteases from complex mixtures is provided hereunder. Essential is that a suitable assay is available that is selective towards the enzyme characteristics sought. For proteases typically a chromogenic, synthetic peptide substrate is used as described in Example 1. Such peptide substrates can be selective towards endoproteases, carboxypeptidases, aminopeptidases or omega peptidases. In Example 11 the selectivity towards a specific tripeptidylpeptidase is described. By choosing the right amino acid residues in the relevant synthetic peptide, proteases with the desired specificity can be selected.
[0278]First it should be determined whether the protease is excreted into the medium, depending on the expression system chosen to produce the protease, it may be excreted or contained in the cell. If the protease is excreted into the fermentation medium, the producing cells or fragments of these cells have to be removed by centrifugation or filtration and the resulting clear or clarified medium is the starting point for further purification. In those cases in which the protease sought is not excreted, the producing cells have to be disrupted to enable purification of the protease. In such cases the collected cell mass is best ground with an abrasive, milled with beads, ultrasonicated or subjected to a French press or a Manton-Gaulin homogeniser and then filtered or centrifuged. In case the protease is hydrophobic or membrane bound, the addition of a non-ionic detergent to solubilise the protease before the filtering or centrifugation step may be necessary.
[0279]After the clarification step, a three phase purification strategy can be applied to obtain the unknown proteases in an essentially pure state. In all or some of these three phases addition of a detergent may be necessary.
[0280]In the first or capture phase the target protease is isolated, partly purified and concentrated. During the subsequent intermediate purification phase most of the bulk impurities are removed and in the final polishing phase trace amounts of remaining impurities of larger amounts of closely related substances are removed and the enzyme is dissolved in the desired buffer. Depending upon the nature and physical properties of the protease at hand, a person skilled in the art is capable of optimising the three phases using slightly modified versions of the different protein binding materials and apply these under somewhat changed conditions. However, in all cases a selective analytical assay is indispensible as it will enable the continuous monitoring of the increasingly purified proteolytic activity. Analytical assays suitable for the purpose include the use of chromogenic peptide substrates as has been mentioned before.
[0281]In the first capturing phase of the purification a strong ion exchange resin of the anionic type is preferably used to apply the clarified and desalted enzyme containing medium. To guarantee binding of the desired proteolytic activity to the resin, three or four different pH values of medium and resin are tested under low conductivity conditions. In these tests the resin is always equilibrated with a buffer of the same pH value and conductivity as the enzyme containing medium. The medium is then applied to the column under pH conditions which has been shown to allow adequate binding of the protease to the resin i.e. none of the desired enzymatic activity can be traced back in the run-through medium. Subsequently the desired enzymatic activity is eluted from the ion exchange resin using a continuous salt gradient which starts with the resin equilibration buffer and ends with this buffer to which 1 mol/liter of NaCl has been added. Eluted fractions containing the desired activity according to the assay are pooled and then prepared for an additional purification step. This additional purification step depends on the purity of the desired enzyme in the pooled fraction: if almost pure, an additional gel filtration step will proof to be adequate; if not almost pure, chromatography over a hydrophobic interaction resin is applied followed by a gel filtration step.
[0282]Chromatography over a hydrophobic interaction resin is carried out by first increasing the salt content of the pooled fraction obtained from the ion exchange resin to 4 mol/liter of NaCl and by removing any precipitate formed. If the resulting clear fraction does not contain the desired activity, this activity is obviously present in the precipitate and can be recovered in an essentially pure state. If the resulting clear fraction still exhibits the desired activity in the assay, then the liquid is applied as such to a phenyl sepharose resin (Pharmacia) equilibrated in this high salt buffer with an identical pH and conductivity. If the desired enzymatic activity binds to the phenyl sepharose resin, the activity is eluted with a continuous gradient of decreasing salt content followed by a salt free wash and, if necessary, with a chaotropic agent. Like before those fractions from the gradient that exhibit activity in the assay are pooled and finally subjected to a gel filtration step. If the desired enzymatic activity does not bind to the phenyl sepharose resin, many of the contaminants will, so the desired proteolytic activity as present in the void volume of the column requires only an additional ultrafiltration step to obtain the activity in a more concentrated form before applying it to the gel filtration column. The gel filtration column does not only remove trace contaminations but also brings the enzyme in the buffer which is required by subsequent use.
[0283]Although this method is generally applicable for the isolation and purification of proteases according to the invention, a more specific isolation technique is described in Example 4. In that Example the isolation of an Aspergillus protease is described by using immobilised bacitracin, a peptide antibiotic known for its selective interaction with various types of proteases.
Example 5
Isolating Proteases by Affinity Chromatography
[0284]An alternative method for purifying small quantities of protease is by affinity chromatography. To obtain the protease in a purified form, a 100 milliliter culture is grown in a well aerated shake flask. After centrifugation to remove any non-soluble matter, the supernatant is applied to a 40 milliliter bacitracin-Sepharose column equilibrated with 0.05 mol/litre sodium acetate pH 5.0. Proteases bound to the column are eluted using the acetate buffer supplemented with 1 mol/litre of NaCl and 10% (v/v) isopropanol (J. Appl. Biochem., 1983 pp 420-428). Active fractions are collected, dialysed against distilled water and applied on a 20 milliliter bacitracin-Sepharose column, again equilibrated with acetate buffer. As before, elution is carried out using the acetate buffer supplemented with NaCl and isopropanol. Active fractions, i.e. fractions displaying the activities sought, are collected, dialysed against a 5 millimol/litre acetate buffer pH 5.0 and then concentrated by means of ultrafiltration with a Amicon PM-10 membrane. To obtain the protease in an essentially pure state, the concentrated liquid is chromatographed over a Superdex 75 column equilibrated with the 0.05 mol/litre sodium acetate buffer pH 5.0 and supplemented with 0.5 mol/litre NaCl.
[0285]Further experiments carried out with the purified enzyme on PAGE may confirm if the molecular weight is in line with what can be expected on the basis of the available sequence data. Final confirmation can be obtained by carrying out a partial, N-terminal amino acid analysis.
Example 6
Properties of a Novel Cysteine Protease from A. niger
[0286]In this Example Aspergillus gene nr 28 was cloned and overexpressed in A. niger as described before. The enzyme obtained was purified according to procedures described in Example 4 and used to destroy trypsin inhibiting activity from soybeans under various conditions. As reference materials papain and bromelain were used. Bromelain was obtained from Sigma, papain was obtained from DSM Food Specialties Business Unit Beverage Ingredients, PO Box 1, 2600 MA Delft, the Netherlands.
[0287]Trypsin inhibition was measured according to the method of Kakade, M. L., Rackis, J. J., McGhee, J. E. and Puski, G. (1974): J. Cereal Chemistry 51: 376-382. Degradation of the substrate N-benzoyl-L-arginine-p-nitroaniline to N-benzoyl-L-arginine and p-nitroaniline was taken as a measure of trypsin activity. Trypsin was obtained from British Drug Houses Ltd and was derived from cow's pancreas containing more than 0.54 Anson Units per gram of product.
[0288]The Kunitz inhibitor for soybeans was also obtained from Sigma.
[0289]The trypsin inhibitor was pre-incubated at a concentration of 2 mg/ml with the above mentioned cysteine protease enzymes at pH 3 in 50 mM Na-acetate buffer prior to measuring trypsin inhibition. Enzymes were added at a ratio of enzyme protein to trypsin inhibitor of 1:100 (w/w). Albumin served as a negative control for the enzymes. Remaining trypsin activity was measured after incubation during 3 hours at 37° C. Results are shown in Table 2.
TABLE-US-00002 TABLE 2 Effects of various cysteine proteases on the enzymatic inactivation of the Kunitz trypsin inhibitor from soybeans. 3 4 5 Remaining TI Remaining TI Remaining TI 1 2 activity after activity after activity after Enzyme Remaining TI pepsin heat treatment heat treatment tested activity (%) treatment at 75° C. at 90° C. Papain 25 55 78 95 Bromelain 30 62 86 99 A. niger 26 26 28 35 Albumin 100 100 100 100 (control) TI: Trypsin Inhibitor activity
[0290]Experiments were repeated in the presence of pepsin during the pre-incubation of cysteine proteases with the trypsin inhibitor. Pepsin was added at final concentration of 1.3 mg/ml. Results are shown in column 3.
[0291]Another series of experiments were conducted to check for heat stability. The cysteine proteases were incubated at 75 and 90° C. during 5 minutes prior to the addition of these enzymes to the pre-incubation with the trypsin inhibitors. Results are shown in columns 4 and 5.
[0292]These results clearly demonstrate the superior activity of these novel cysteine proteases from Aspergillus niger over currently available cysteine proteases for the inactivation of trypsin inhibitors in animal feed.
Example 7
Exo-Peptidases Promoting Cheese Ripening and Cheese Taste
[0293]The amino-peptidases encoded by genes nr 20 and 54 (see Table 1) were overexpressed in A. niger according to methods described earlier. Purification of these enzymes was carried out according to procedures as described in Example 4. The activity of the purified enzyme samples was determined at pH7.2 in an aqueous phosphate buffer (50 mM) containing the para-nitro anilide derivative of a number of hydrophobic amino acids (3 mM) as the substrate. The conversion of the substrate by the amino peptidase was determined by monitoring the change in optical density at 400 nm as a result of substrate conversion, using a solution not containing the enzyme as the reference. Activity (A) was calculated as the change in OD per minute and expressed as e.g. Phe-AP, Leu-AP or Val-AP units, depending on the substrate used. Normal cheese milk was inoculated with starter culture of the Delvo-tec® DX 31 range (DSM Food Specialties Delft, The Netherlands) to obtain a Gouda-type cheese and coagulating was executed with an average dosis of coagulant (50 IMCU per liter of cheese milk). In addition, 25 Phe-units of each exo-protease was added to two experimental cheeses whereas the control did not contain either one of the exo-proteases. Cheese making parameters were used conform the procedure applied for semi-hard cheese for both cheeses. A difference was noted in terms of flavor and aroma development between the experimental cheeses and control cheese to such an extent that the experimental cheeses has obtained most of its organoleptical properties after three (3) weeks whereas the control cheese has obtained a similar qualification after six (6) weeks. The level of free amino acids after three weeks was shown to be twice as high in the experimental cheeses; after six weeks of ripening the levels were comparable again. Amino acid analysis was carried out according to the Picotag method of Waters (Milford Mass., USA).
[0294]These data suggests that the product is ready for sale three weeks earlier without decreasing the keeping quality of the cheese. The organoleptic character of the experimental cheeses differed from the control to the extent that the bland cheese flavor with a slight tendency to bitterness of the control cheese was overcome in the experimental cheese in the presence of the amino-peptidase. The texture of the cheeses was found to be somewhat smoother as well.
Example 8
Novel Specificity of a Protease Encoded by Gene 55
[0295]As explained earlier, certain proteins can resist enzymatic hydrolysis as the result of specific amino acid compositions or specific tertiary structures. In such cases the quantity of peptides that can be solubilised from protease resistant proteins can be dramatically improved by using proteases exhibiting novel specificities.
[0296]Beta-casein is a protein with very limited tertiary structure but with an extraordinary high level of proline residues. Many proteases have difficulties in cleaving proline containing sequences so that the hydrolysis of beta-casein with commonly available proteases yields a hydrolysate that is relatively rich in large, protease-resistant peptides. The latter resistant peptides can attribute to a number of undesirable properties of the hydrolysate. For example, it is well known that these larger peptides have a relatively strong effect on allergenicity and bitterness. Moreover, these peptides withstand a further degradation into free amino acids so that in certain processes the occurrence of these large, protease resistant peptides are synonymous with yield losses. Therefore, the availability and use of proteases that are capable of cleaving the protease-resistant parts of the proteins, translate into serious technical and economical benefits.
[0297]Beta-casein represents one of the major casein fractions of bovine milk. The protein has been well characterised in terms of its amino acid sequence and is commercially available in an almost pure form. As such, beta-casein offers an excellent test substrate for studying the relationship between enzyme cleavage sites and the length of various peptides formed during enzyme hydrolysis.
[0298]This Example demonstrates that despite the broad spectrum cleavage character of the endoprotease subtilisin, the addition of a very specific enzyme like a prolyl endopeptidase as encoded by gene 55 (see Table 1) has a major impact on the size of the beta-casein fragments formed.
[0299]Beta-casein from bovine milk (lyophilised, essentially salt-free powder) with a minimum 90% beta-casein was obtained from Sigma. Subtilisin from B. licheniformis (Delvolase®, 560 000 DU per gram) was obtained from DSM Food Specialities (Seclin, France). The proline-specific endoprotease as encoded by gene 55 was overexpressed in A. niger and purified using procedures described in Example 4.
[0300]Beta-casein powder was dissolved at a concentration of 10% (w/w) together with 0.1% (w/w) Delvolase® powder in a 0.1 mol/liter phosphate buffer pH7.0. After an incubation of 24 hours at 45° C. in a shaking water bath, the reaction was stopped by heating the solution for 15 minutes at 90° C. To one half of the solution (1 ml containing 100 milligrams of beta-casein) 100 microliter of the proline-specific protease was added and the reaction was continued for another 24 hours at 45° C. After another heat shock at 90° C., samples of both the Delvolase® and the Delvolase®+proline-specific endoprotease treated beta-casein material were analysed by LC/MS equipment to study the precise peptide size distributions in the two samples.
LC/MS Analysis
[0301]HPLC using an ion trap mass spectrometer (Thermoquest®, Breda, the Netherlands) coupled to a P4000 pump (Thermoquest®, Breda, the Netherlands) was used in characterising the enzymatic protein hydrolysates produced by the inventive enzyme mixture. The peptides formed were separated using a PEPMAP C18 300A (MIC-15-03-C18-PM, LC Packings, Amsterdam, The Netherlands) column in combination with a gradient of 0.1% formic acid in Milli Q water (Millipore, Bedford, Mass., USA; Solution A) and 0.1% formic acid in acetonitrile (Solution B) for elution. The gradient started at 100% of Solution A and increased to 70% of solution B in 45 minutes and was kept at the latter ratio for another 5 minutes. The injection volume used was 50 microliters, the flow rate was 50 microliter per minute and the column temperature was maintained at 30° C. The protein concentration of the injected sample was approx. 50 micrograms/milliliter.
[0302]Detailed information on the individual peptides was obtained by using the "scan dependent" MS/MS algorithm which is a characteristic algorithm for an ion trap mass spectrometer. Full scan analysis was followed by zoom scan analysis for the determination of the charge state of the most intense ion in the full scan mass range. Subsequent MS/MS analysis of the latter ion resulted in partial peptide sequence information, which could be used for database searching using the SEQUEST application from Xcalibur Bioworks (Thermoquest®, Breda, The Netherlands). Databanks used were extracted from the OWL.fasta databank, available at the NCBI (National Centre for Biotechnology informatics), containing the proteins of interest for the application used.
[0303]By using this technique as a screening method only peptides with a mass ranging from approx. 400 to 2000 Daltons were considered suitable for further analysis by MS sequencing.
[0304]Angiotensin (M=1295.6) was used to tune for optimal sensitivity in MS mode and for optimal fragmentation in MS/MS mode, performing constant infusion of 60 μg/ml, resulting in mainly doubly and triply charged species in MS mode, and an optimal collision energy of about 35% in MS/MS mode.
[0305]In the sample digested with Delvolase alone, the LC/MS/MS analysis identified 40 peptides covering various parts of the beta-casein molecule. Together these peptides accounted for 79% of the total beta-casein sequence. Different retention times of the peptides on the C18 column could be traced back to peptide lengths ranging from 2 to 23 amino acid residues. Together <15% of the peptides found were smaller than 6 amino acids. The sample digested with Delvolase® and the proline-specific protease also generated a large number of identifiable peptides from beta-casein. Together these peptides covered >50% of the total beta-casein protein sequence. In this sample the peptide size distribution was remarkably homogeneous, as the peptides ranged in length only between 2 and 6 residues. The results show that in the hydrolysate made with the proline-specific protease contain a large fraction of di-, tri-, up to 6 M peptides, showing the distinct beneficial effect of the co-incubation with an endoprotease featuring an unusual specificity. It is also clear from these experiments that the endoprotease according to gene 55 encodes an endoprotease that cleaves the peptide chain at the carboxy terminus of the proline residue.
Example 9
The Selective Release of Specic Amino Acids to Promote Flavour Formation
[0306]Free amino acids like leucine and phenylalanine have not only been implicated in Maillard reactions but also as precursor for desirable aromas in various food fermentations. To promote the formation of such aromas in food fermentations or during the heating, roasting or baking phase of food, it would be advantageous to incorporate into these products a protein hydrolysate that contains relatively high levels of these specific amino acids in a free form. In this Example we describe the production of yeast extracts selectively enriched t in leucine and phenylalanine. This enrichment is obtained by combining an endoprotease with a cleavage preference for a selected set of amino acid residues with an exoprotease favouring the release of a similar set of amino acid residues. The preference of the endoprotease should match with the preference of the exoprotease used. For example we have established that the aminopeptidases encoded by genes 20 and 54 (see Table 1) feature a definite preference for releasing leucine and phenylalanine residues which matches with the cleavage preferences of thermolysin. The carboxypeptidases encoded by genes 23 and 24 have a preference for releasing arginine and lysine residues which matches the cleavage preferences of trypsin. Carboxypeptidase encoded by gene 5 features a highly unusual preference for releasing glycine which could be combined with certain endoproteases present in papaine. The carboxypeptidase encoded by gene 51 is capable of removing glutamate residues which matches the glutamate specific protease encoded by gene 43.
[0307]The endoprotease thermolysin (commercially available as Thermoase) C 180 from Daiwa Kasei KK (Osaka, Japan) is known to cleave peptide bonds at the amino terminal side of bulky, hydrophobic amino acids like Leu and Phe. To liberate the thus exposed amino acids from the newly formed peptides, we used the amino-peptidases encoded by genes nr 20 and 54 (see Table 1). These genes were overexpressed in A. niger according to methods described earlier and purification of these enzymes was carried out according to procedures as described in Example 4.
[0308]To release as much leucine and phenylalanine as possible without concomitant release of undesired amino acids with this combination of enzymes, it is evident that the conditions used during enzymatic hydrolysis should be carefully selected. Moreover, the yeasts own endogeneous (and probably aspecific) proteases have to be inactivated. After a number of test incubations, a protocol was worked out that leads to a surprisingly selective and effective release of leucine and phenylalanine from the yeast proteins using these two new enzymes.
[0309]To inactivate the yeasts endogeneous proteases, the yeast suspension was kept for 5 minutes at 95 degrees C. Then the suspension was quickly cooled down to the required temperature and the pH was adjusted to 7.0 using 4N NaOH. The yeast, the thermolysin and one of the aminopeptidases were all incubated simultaneously under the following conditions. After the heat shock, the pH of the 2000 milliliters yeast suspension was adjusted to 7.0 after which 680 milligrams of Thermoase were added and, after stirring, the purified aminopeptidase. The mixture was incubated with stirring at 50 degrees C. for 3 hours and centrifuged. To stop all enzymatic activities the pH of the supernatant was adjusted to 4 and subjected to another heat treatment of 45 minutes at 95 degrees C. After another centrifugation a sample for amino acid analysis was obtained from the supernatant. Precipitated or non-dissolved matter was removed by centrifugation for 15 minutes at 3500 rpm in an Hereaus Megafuge 2.0 R centrifuge. Supernatant was removed and kept frozen at -20° C.
[0310]Samples of the supernatant, were analysed for amino acid content according to the Picotag method of Waters (Milford Mass., USA) immediately after thawing.
[0311]In the amino acid analysis Trp and Cys values were omitted And Asp and Asn values were summed as one value. According to the data obtained, in the resulting hydrolysate the ratio between alanine and leucine (21.3:11.7) was 1:0.5 Commercially available yeast hydrolysates typically exhibit alanine versus leucine ratio's of 1:0.3.
[0312]In a second experiment a yeast extract was prepared that was enriched in free glutamate. To achieve this, use was made of an endoprotease exhibiting a preference for cleaving at the C-terminal end of glutamate residues (encoded by gene nr 43 in Table 1) and a carboxypeptidase (encoded by gene nr 51 in Table 1) capable of removing these glutamate residues thus exposed. The endoprotease encoded by gene nr 43 and the carboxypeptidase encoded by gene 51 (see Table 1) were overexpressed in A. niger according to methods described earlier. Purification of these enzymes was carried out according to procedures as described in Example 4.
[0313]The essential role of free glutamate in a number of aroma forming processes is well documented and MSG, the sodium salt of glutamic acid, is recognized as the single most important taste enhancing component.
[0314]In this Example the pH of the 200 ml heat shocked yeast suspension is adjusted to 8.0, then the purified enzyme product encoded by gene 43 is added and the mixture was incubated for 4 hours at 50 degrees C. Then the pH was lowered to 5.0 and the suspension was centrifuged. To 100 milliliters of supernatant the purified gene product of gene 51 is added. Incubation with this carboxypeptidase took place for 30 minutes at 50 degrees C. with continuous pH adjustments. After stopping the enzyme incubation by a heat treatment of 5 minutes by 95 degrees C., the material was again centrifuged (see above) and a sample was obtained for amino acid analysis.
[0315]According to the amino acid data obtained (see above), in the resulting hydrolysate the ratio between_alanine and glutamate (30.0:48.7) was 1:1.6. Commercially available yeast hydrolysates typically exhibit alanine versus glutamate ratio's of 1:1.
Example 10
Flavour Evaluation of Yeast Hydrolysates Enriched in Specific Amino Acids
[0316]To prove that a protein hydrolysate enriched in specific amino acids according to the invention can generate specific aroma's, a number of experiments were carried out with the yeast hydrolysates described in an earlier Example. To that end larger portions of these hydrolysates were prepared and lyophilised. The performance of the resulting powders were compared with the performance of a commercially available yeast extract (Gistex LS, obtainable from DSM Food Specialties, Delft, The Netherlands) in a standardised mixture under several reaction conditions. The standardised mixture consisted of one of the hydrolysates, base mixture and water.
[0317]The base mixture contained 22 grams of Maxarome Plus Powder (a specialised yeast extract with a high content of natural nucleotides, also obtainable from DSM Food Specialties), 29.2 grams of glucose, 9 grams of REFEL-F fat (hydrogenated soy oil, obtainable from Barentz, Hoofddorp, The Netherlands) and 0.2 grams of calcium stearoyl lactylate (emulsifier, obtainable from Abitec, Northampton, UK) thoroughly mixed in a mortar.
[0318]All standardised mixtures contained 5 grams of yeast hydrolysate powder (i.e. either the leucine or the glumate enriched material or the commercial yeast extract), 3 grams of the base mixture and 3 grams of water. After thorough mixing, these three slurries were subjected to different heating regimes i.e. either 65 minutes at 90-95 degrees C. in a reaction vial (liquid reaction) or dried at 20 millibar at 120 degrees C. in a vacuum oven (vacuum roast reaction) or heated in an open reaction vial at 120 degrees C. for 10 minutes after the dissipation of all water (roast reaction).
[0319]After the heat treatment all three products had assumed colours ranging from dark brown to almost black. In case of the vacuum roast reaction only the light coloured top layers were used. Taste evaluation of the heated products was carried out by grinding the blackened cakes into fine powders and dissolving these powders to a concentration of 2% (w/w) in water containing 0.6% (w/w) NaCl. The observations of the taste panel are specified in Table 3.
TABLE-US-00003 TABLE 3 Reference Leucine Glutamate Liquid Bouillon, Cold tea, slightly More bouillon, slightly roast flowery, yeasty meaty, yeasty Vacuum Burnt, fried Astringent, beans, Burnt, bouillon, roast potatoes yeasty yeasty Roast Dark roast, Less roast, flowery, Roast, more bouillon, bouillon, umami umami more umami
Example 11
Non-Allergenic Whey Protein Hydrolysates Formed with Tripeptidylpeptidases
[0320]The dipeptidylpeptidases encoded by the genes 19 and 55 as well as the tripeptidylpeptidases encoded by the genes 4, 9, 10, 12, 26, 35, 46, and 50 (see Table 1) may be overproduced as described and may be purified according to the methods provided in Example 4. After purification the pH optimum and the temperature stability of each individual enzyme may be established by any of the methods available and known by the skilled person. Furthermore, the specificity of each individual enzyme may be determined using the methods outlined in Example 1. The selectivity exhibited by tripeptidylpeptidases is illustrated in the following experiment.
[0321]The enzyme encoded by gene 12 was overproduced in an Aspergillus niger host cell and purified by procedures described in Example 4. The enzyme thus obtained was incubated at pH 5 and 50 degrees C. with different synthetic chromogenic substrates i.e. Ala-Ala-Phe-pNA and Ala-Phe-pNA (both from Bachem, Switserland). The incubation with the Ala-Ala-Phe-pNA substrate led to a significant increase of the absorbance at 410 nm whereas the incubation with Ala-Phe-pNA did not. This observation clearly demonstrates that tripeptidylpeptidases cleave off tripeptides and do not exhibit aminopeptidase activity that can lead to an undesirable increase of free amino acids.
[0322]Moreover, the enzyme encoded by gene 12 shows favourable enzyme stability characteristics as shown in the following experiment. Four samples of the enzyme were incubated at pH 5 for one hour at 0, 40, 50 and 60 degrees C. respectively. Then each enzyme sample was incubated with the above mentioned Ala-Ala-Phe-pNA substrate in a citrate buffer at pH5 and the residual activity in each individual sample was determined by measuring the increase in absorbance at 410 nm. With the 0 degrees C. sample showing 100% activity, the 40 degrees sample showed 96% residual activity, the 50 degrees sample 92% residual activity and the 60 degrees sample 88% residual activity.
[0323]In a typical process aimed at producing a hydrolysate with a high proportion of tripeptides, whey protein (WPC 75) may be dissolved/suspended in a concentration of 100 grams of protein/liter, in an aqueous medium having a pH of 8.5. The first enzyme incubation is with the broad spectrum endoprotease subtilisin (Delvolase®, 560 000 DU per gram from DSM). After a predigestion of the whey with this enzyme in a concentration of 0.5% enzyme concentrate per gram of protein for 2 hours at 60 degrees C., the mixture is heat-treated to inactivate the endoprotease used. Then the temperature is adjusted to 50 degrees C. and the tripeptidylpeptidase is added and the whole mixture is incubated until the desired level of tripeptides is reached. Further processing steps of the hydrolysate thus obtained depend on the specific application but may incorporate microfiltration or centrifugation followed by evaporation and spray drying.
Sequence CWU
1
17112520DNAAspergillus niger 1cgcaggcgtc cgttgcgccg cgaaaacctg ccgagtgggc
cgtttaggct ttgggtctcc 60ccacgatgta agcataatca ttctgtgcct gagtgtgaat
tctcctgttg gaggctgcat 120cttaattctt aactgcatga aaagcacttg ggtgctattt
tctttttcct ttctttcttt 180tccgtgttca tttccattcc cttgctcttc ttctttgtgt
cgacatttac aaatcacatt 240tttcttatac tttcttttct tcacctcgtt tcttcctatt
cactctctgt gttcagcatt 300cgttatcagc actttatctt ttgctcgtct cttttatctt
cacttgtttg tgcctttcca 360ctagcaatct atcgtttgat ctttctagag cattgtcttg
attgtgtcat tctgtcattg 420actccggcta tgaaatatta ttctcaatct gcctaaaacc
aaattctact ctatcactac 480acatttgtat cacctgatct ggctgagata ggagagtccg
gcatctcatc gtctgcatca 540gacaattgcg ataaattcat tgcttgcacc tgttattgat
tcttccaagt tatgcatctc 600ccacagcgtc tcgttacagc agcgtgtctt tgcgccagtg
ccacggcttt catcccatac 660accatcaaac tcgatacgtc ggacgacatc tcagcccgtg
attcattagc tcgtcgtttc 720ctgccagtac caaaaccaag cgatgctcta gcagacgatt
ccacctcatc tgccagcgat 780gagtccctgt cactgaacat caaaaggatt cccgttcgtc
gtgacaatga tttcaagatt 840gtggtagcgg aaactccctc ttggtctaac accgccgctc
tcgatcaaga tggtagcgac 900atttcataca tctctgtcgt caacattggg tctgatgaga
aatctatgta catgttgctc 960gacacaggcg gctctgatac ctgggttttc ggttccaact
gcacgtccac accctgcacg 1020atgcacaata ccttcggttc ggacgattct tcgacccttg
aaatgacatc ggaagagtgg 1080agtgtgggct atggaactgg gtctgtcagc ggcttgctag
gaaaagacaa gctcacgatt 1140gcaaatgtca ctgtacgcat gactttcgga cttgcttcca
acgcatcgga taacttcgag 1200tcgtacccaa tggacggcat tctcggtctc ggtcgaacca
acgatagttc ctacgacaac 1260ccaacattca tggatgccgt tgcagaaagt aacgttttca
agtcgaatat cgttggcttc 1320gccctttcac gtagccccgc caaggatggc acggtcagct
ttggcactac tgacaaggac 1380aagtacaccg gcgatatcac ctacaccgat accgtcggat
cggacagcta ttggcgcatt 1440cccgtggacg atgtctatgt tggcggcact tcatgcgatt
tctccaacaa atcagccatc 1500atcgataccg gaacttctta tgctatgctg ccttcaagcg
actcgaagac gctgcacagt 1560ctcattcccg gcgccaaatc ttcggggagc taccacatta
ttccgtgcaa cacaactact 1620aagctacaag tggcattctc tggtgtgaat tacaccatct
cgccgaagga ctacgtggga 1680gcaacttcag gttctggatg cgtttcgaac attatcagct
acgacttatt tggtgatgac 1740atctggctcc tgggtgacac gtttctcaaa aatgtgtatg
ctgtgtttga ctacgatgag 1800ttacgggtcg gatttgcaga gcgttcctcg aacaccacct
ctgcgtcgaa ctctacgagc 1860tctggaacaa gcagcacctc gggatccact acaacgggca
gctcaacgac tacgacgagc 1920tctgctagct ctagtagttc atctgatgct gaatcaggaa
gtagcatgac cattcccgct 1980cctcagtatt tcttctctgc tctggcgatt gcttccttca
tgctttggct ctagttaacc 2040gcatcttact cgacgcctga acctcgggaa acatatgcat
tatttacaca tgctgctgat 2100ttgtatttgc atatattctt cgagcctgga cggcgtgcgg
gtcatattac cttacattcg 2160aagtccttct ctaatcaatc aacatttatt cttactccac
cagttctggc tcgcaattaa 2220ccctgtctaa gaaaaagttg gtatagaaca tggcatccac
tacctggaac attcaaagaa 2280ccttgtccgg gatcagtgtg tatgacttcg ggtacgattc
tgacatgaca gttagcgtcc 2340atcctgagga ttcatcctga tctccttacc tagtatggac
ttatcaaagt ccttgacgct 2400attgtaccct cccacagcca tcaggatatc gaaatcggcc
aacagcgact tcatcacatg 2460gcgaacaccc tcctcaccca tgatactgag gccccagatc
cagagccggc cgacgaaaac 252025001DNAAspergillus niger 2taggaaaatc
agaggcgaca atttgctccg atactggata agtaccatcg gtcacgaaat 60tcagcaccga
ctggcttatg tcctgctccg ataccttagc ggacattatg ttagtagttc 120taagcaagag
cccttggttc ttggtaatcg gcggatcaaa aacgtaaaga agacaaacag 180aagcgccaca
ctaggctctg cctccctctt acagaagatc tgccaggtat cgatccacaa 240agatatacca
aggactccaa tgcagcagat cgcaaccgaa agggaaggcg agctgtcaac 300accgccatgt
gacttcaccg ctcaccgcct tagctgcgat gggcaagacc acgaacgatc 360tcgctcttcc
ccagggccag actgaggccg attatatgat ttttttcttt cttttccgtg 420tgtctcgtct
gctgctatat tcttattttt ctgttcggta aagatacctc ctaagaatag 480acacgggttg
tttttttgtg aattatctgt tggtgtggtg ctctctatcg aaccggaaac 540ctgaactcca
ctctgaaccg ttacagttgg agggtaacct tcgttcagcg gccaactgtg 600tgacctcaaa
attcctgaaa catcataatc ggcgtcaagc agagcccatc gtgttgtctc 660agttactatt
gagaagcctg attcgggcaa acgcttctcg atccatgtga gtcatgcttg 720taccagccct
gaactccatt gaataaaaaa aaagcaagaa aagagattgc ccctccgtcg 780ttttctgtat
taaacatcca atcctcaacc ggcagcctta cattggctgc cacagatttg 840tgcctccata
cagcatgctt cgtggtcttc gtgatgtcgt attattacaa tttgcaatcc 900ccttgttctt
gctattggta tttcaaacat ggaatgggtg cttttcccgt ttcaattccc 960cgcaccctga
acctcaagca ttgctagctt ctctaaaatg aataaaactg ttatacttgc 1020ttcctaaaag
tttgcgcttc tggcctatgg attactaaca gcattttaga ttatcgctac 1080ggggtgtgat
cacaggattt ggttctaaat cacatttcca gagaccattg agcaaaatgt 1140catctactca
aaagagccat ttcaagctac tccagaagtt caaaccggag tactcgccta 1200gcgagtttgc
tcagtatgag tcggagagaa caggcatgag ggtagtggtc attgaccaaa 1260aaggacccaa
agtcacaggt tattttgttc tagccacaga gattctcgat gattcaggtg 1320ctcctcacac
gttggagcac ttgtgcttta tgggctcgcg gaactataga tataagggct 1380tccttgacaa
gctagcaaca cgtgtttatt cgagcaccaa tgcctggacg gccacagacc 1440acacggccta
caccttggac acagcaggct gggaagggtt cgctcaaatc ttgcccgtgt 1500acctagagca
tgttatagct ccaacactga cagatgaagg gtgctatacc gaagtgcatc 1560atattgatgg
cgctggagac gacgctggag tcgtctactc ggagatgcag ggtgtgcaga 1620ataactctgc
agagttaatc gatctaaccg ctcgtcgatt gacttacccg catggtgtag 1680gttttcgcta
cgagacaggc ggtatgatgg agcagctccg cgtcctcacc gcggaccgta 1740tccgagcgtt
ccatcgtgag atgtaccagc ccaagaactt atgcctaatc atcacaggcg 1800aagtagatca
ccagaacatg ctggagacct tggacaagtt cgaagatact attctagatg 1860tcattcccag
tcctgattca cctttcaaga ggccgtgggt agattccaag caggcgccgc 1920cattggagaa
gtccattgtc cagactgtgg aatttccgga agaagatgaa tctttcgggg 1980agatagaaat
tagattcctc ggtccggact gtaccgaccc tgttcaaagt gagtgttccg 2040ctgtcctcat
ttcgaagata tacttactct gttatagccg gggctgtcaa tgttgcattg 2100ctgtatctgg
ccggttcatc tgcttctcta ttggataaca tcctggttga gaaggagcag 2160ctcgccagtg
ctgtctatta tgctaccgaa gatcatccca gcattgagat ccgcttcaca 2220ttaaccagtg
tggagacaga gaaactcgcg aaggtagagc aacggttttt cgaagtgctc 2280aaggacgcta
tggagaaaga tttagacatg aggtatatca aggagtgcat tgaccggcaa 2340agacggacct
ggaagttctc taccgaaagc tccgcctctt cctttgcgga gtacgtgatc 2400tcggattttc
ttttcggaaa gagagacgga tcgactatgc ttgatgttgc gaccttgcaa 2460gagtacgacg
tgctggagaa gtggagtgaa gaacagtggc gcagttttat caaaacatgg 2520atttctgatg
ccaaccatgt cactatcctt ggtgttccgt ccgttaagat gtctgacaca 2580ttaaagaagg
aggaggaagc tagagtcgca gagcaaaaga agcgcttggg tgatgagggg 2640ctgaagaagt
tggccgacaa gctggaaaaa gctaaagctg aaaatgacaa ggagatcccc 2700aaggagatgc
tggagaggtt ccaaatccct ggaatagagt ctatccattt cgtggacact 2760actacagcca
ggtctggtgc agccctcgat gccgggcgcc catcccacaa ggcgcaaaaa 2820ctggtggatg
ctgatggctc tgatctgccc ttgttcatcc atttcgagca tatccccagt 2880agcttcgtgc
agctctccct cctcatctcg gcacaggccg tacctgtgca gcttcgtcca 2940ctgctgtctg
tgtatactga ggcattcttc aacctgcctg tcaaccggaa cggggaaacc 3000atcaactttg
agcaggtggt tgtcgagttg gaaagggata ctgttggcta ctccatggaa 3060ggagctagaa
gcctaggaaa ctcggagatg ttgcggatct cattccaggt ggagcttgag 3120aagtatcaca
cggcgatcgc atggatccag gaactttcct ggaactcgat tttcgatgtc 3180gagcgactcc
gagcgattac cagtcgactg ctctccgatg tgcccgattc caagcgtagt 3240ggcgacgaca
tgctcgcggc tgttcatgtg atggtccact atgcagcaga gtctattgtt 3300cgggctcgga
gcaccttggt gaaggcgcgt tatttgaaac ggatcaagaa gcaattagca 3360gaagagccga
agtctgtcgt tgcgcggatg gaagaaatca gagatgcgct tttccgtttc 3420gagaacatgc
gagtcttagt tatcgctgac ctggagaaac ttcaaaaccc tgtgtcagca 3480tggaaaccat
ttgctgagcg tttgggtgca ggtgcccctc tacagcctat cacgactaga 3540agaccgttgc
tcagtgaggc aggccagaag ttgggcggta agtcgtatgt ggttcctatg 3600ccgacgattg
attcatcgtt cgcatatgct accgcacgtg gtttggattc ttatgatgat 3660ccaagacttc
ctgccttaat ggttgcaatt gcatacatga acgcggttga gggtcccctc 3720tgggttgcag
ttcgaggcaa gggtttggca tatggcacga actttgccta taacattgat 3780accggattcg
tcaacttcga cgtttaccgc tcccccaacg cccataaagc cttcgactcc 3840agcaagcaga
ttgttgagga tcacctctct ggtgcgatgc ccttcgatcc cttgatgctg 3900gagggttcca
ttagcagcat tgtggtaagc tttgcgaatg aacagtcgac aattggtagc 3960gcagcctcag
gcagtttcat ccgacaggtg attcggcgcc tgcctagcga ctacaaggag 4020cgggtgctca
agcaggtgcg ggctactagc gttgatgacg tgaaaggcgc tctgaaggac 4080atcattctgc
ctttgtttaa cccgtccacg gccaatatcg tggttacctg cgctacagtg 4140cttgaggagg
tttgtcattc catgaagaaa ttatgatctt cttgtgtatc atttactaac 4200tgtcggttta
gactatcaag gaaggtctcc aggcatcggg attcacgcct gcggtgcagc 4260cactcaaaga
attcgaagat gactatgggc tgaaggtcgg cgatgacgag gacgaggagt 4320ccgacgatga
cgacgatgag tatgaaaccg gatctgaaga tgaagatgac agtgatgaag 4380acatggagga
tgacgaagat gatgagtgat gcaatctata cgacaacctc tagacatgac 4440aagatttatc
tgagccagtt cctggataac acctaggtcg aaagaccagc tacccctggg 4500ggcccagata
tgccgacccg tgtgatctgt attgttagag atgtctccaa ctagcagacg 4560cgcaatgatt
tttgatgtta atgatatatc gtacaacata agtggttaac caccacaaat 4620ctgcacctaa
atcttagttg tgattatcgg tacctaccaa acccgagtaa acgttgccag 4680aagtttatga
aaaagctctc gttcttctta tttcattgaa tgttgtaata aaaagtgcta 4740gcagatgctt
tcgctatcgg gcccactact atgattaacc tgaagcttag tctcttaggt 4800gacataacct
gttctacaca ggctgccatt atatttgcaa cacacagctt ctatctctga 4860cctttgtgag
aataatacaa ttactagaaa ccccaggaga tataaataca ctctactcct 4920accgaactaa
cacccagatc gacaaattat aaaccaaacc acgtgacaat atgatatcta 4980tatatgaata
tgtataaggt a
500133850DNAAspergillus niger 3gcgcggggtt cctgcatgtc tttcatggtt
gggaggtata tgtacatgta atgccgttgg 60gttctggata cccacaataa agtgtttcta
ctattacttt tgcattgtta gactttgaag 120gctgctctgt aagggtccat tccgttggag
tcaaacaacc gtctgtccgc gggcatatgt 180ctcagtctga ggtgagaact gcaatataca
ctccaagctt tctcaatctg atgcacagat 240tgcaaacatc tcccttgctg tgtggtaaag
tctccctttc tcggtgaacc ttcttcgggg 300tcgcaactga ttccattgac cctcaccatc
tgtcgaattg ttcccgaaag ctcgcgtcca 360aataggcttt acggacgcca cagctgctcg
aggccatctc caagggggaa tgaacaatgg 420aatggtaagc cgctaagaaa ggggaactgt
cggttctggt cgattggcag ggtcagggca 480gtgcttgggc tgttcctctg gccctccctt
gcttcaagct ggccccgatt tggcagctcg 540tgaagctcac cagacattca gccagccaga
tgactggcac ctttccgctg ggcataaatt 600ggccctggca gccatgtcat gccaatcttg
gctccaggac cagtcattcc ttttcttgtt 660cctgtcaaac agatcaagtc ctcgaggatg
ggagctcttc agtggctgtc catcacggct 720gctgcggcct ccgcagtgtc agccttgacc
ccggagtaag tatctccaat catttggaac 780tggcccatat tgtgcatagc taaccagctt
acctgcatag gcagatgatc ggtgccccac 840ggagaaccga agttatacca aacccctccg
gtgtatgccc attgccaggt ccagccttac 900aaagaagcgt cgtctgctga cacgagaagg
acaccggtct attctcgacc tcccaatggt 960cgtttgacac tcattctgag agcacctggt
ggagcttgat cgacctccaa tcgggcaaga 1020ccaccactct caccgatgat agcgatatcg
aggagatcat ctggcttggc tctgacaatt 1080ctacgctcct ctacatcaac agcaccaacg
cgcaggttcc cggtggcgtg gagctgtgga 1140ttgcggactc ttctgacttt gcaaatgcgt
tggttcaggc ctttaacgat gcctctgcag 1200actagtgcta atcctacttg gtgcagttac
aaggcagcct ctctctccgc cggttttctc 1260ggcatcaaat caaccgtgac agattccggc
gacgtgcatt tcatccttcg tggaaagtcc 1320tatcccaacg gaacggcata caatgatcag
ctcgccgaga cctatcccag tacagcccgc 1380atctacgaca gcatctttgt gcggcactgg
gacacttacc tgaccaccgc ctcccacgct 1440gtattctccg gtaccctgca aagctcgacc
agcgacgacg gcaatgttca atatacctct 1500tcagggggat tgaccaacct ggttaaccca
gtcaagggtg ccgaaagccc attccctcct 1560tttggaggca acgacgacta tgacctctcg
cctgacggca aatgggttac cttcaagagc 1620aaagcgccag agctgcctct tgctaacaac
acggctgcct atgtctatct cgtcccacac 1680gacggctctg cgactgcctt tgccgtcaac
ggccctgata gtcctgcaac cccggaggga 1740gttgaaggag aatccaacaa tcccgtgttc
tcccctgata gcgacaaaat agcgtacttc 1800caaatggcta ctaatacata cgagtcggac
cgcaacgtgc tatacgtata ctccatcgcc 1860gatgacacca tcactcccct tgcaaaggac
tgggaccgat cgcctagctc cgtgacatgg 1920gtcgatggag acaacctcgt cgtggcaagc
caagatctag gacgaaccag acttttcgcc 1980atcccaggcg atgcagggga cgacttcaag
cccacgaact tcaccgacgg cggctccgtg 2040tcggctcaat acgtcctatc caactctacc
ctcctcgtca cgtccagcgc cttctggaca 2100agctggagcg tctacaccgc cagccctgac
gagggcgtga tcaacacact ggcctcagcc 2160aacgagatcg accccgagct tagcggcctt
agttcctccg actttgaaga gttctacttt 2220gacggcaact ggactaccgt aagtctatcc
ctccttgccc ccaccaccac atcacaaaca 2280tactaaactc accgcagctc caaggatgga
tcacctaccc ccaagacttc gactcatcca 2340agaaataccc cctcgccttc ctcatccacg
gcggccccga agacgcctgg gcggatgaat 2400ggaacctgaa atggcactcc aaggtcttcg
ccgaccaggg atacgtcgtc gtccagccaa 2460accccacagg aagcaccggg ttcggccagc
agctcacaga cgctatccaa cttaactgga 2520gtacgccatt ccctatcccc aaactcccct
cttaaacata cagctaacaa atgaaataac 2580agccggcgcc gcctacgacg acctaaccaa
agcctggcaa tacgtgcacg atacctacga 2640cttcatcgac acggacaacg gcgtcgccgc
gggtcccagc ttcggcgcgt tcatgatcac 2700ctggatccag ggcgatgact ttggacgcaa
gttcaaggcg ctggttagcc atgatggtcc 2760gttcattggc gatgcgtggg tcgagacgga
tgagttatgg tttgttgagc atgaggtgag 2820tggaccaagc caaacccccc ttttctcccc
ttacaccatt acccctatac aaatatgatg 2880attctgaccg tgtatagttc aacggcacct
tctggcaagc gcgcgacgca ttccacaaca 2940cggatccatc cggccccagc cgcgtcctcg
catacagcac cccccagctc gtcatccaca 3000gtgacaagga ttatcgcata cctgtggcaa
atgggattgg actgtttaat acgctgcagg 3060agaggggcgt gcccagtcgg tttttgaatt
tcccggatga ggatcattgg tatgttcata 3120cccttttctt cccccttttt tctcccatga
ttatgggtgt tgtggatgct gatgtagcta 3180tgtgtgtgtt tagggtcacc gggcaagaaa
acagcctcgt ctggtatcag caggtgctgg 3240gatggattaa tcggtattct ggggtgggag
ggtcgaatcc tgatgcgatt gctttggagg 3300atacggtgaa tccggtggtg gatttgaatc
cttgatcatt ccctgttgct cgttactact 3360agtcagttat gatcatgttc tggctgggtc
ccaggataga tggacggtta ttactggtcg 3420ttatattctg tttgcagtgg ttactagttg
gtagatcagt tacgagatga tgtatgagac 3480gacagaaaga cttatgctta tgttgggagc
tgcttcattt gtatatcaat ttattcttgt 3540agtgattgat aactaactac tgtctgactg
tctgtcaacc gctatactaa cattatacca 3600accaaccaac caatcaaatc aatttctggt
tctttcttct cttctgtttc ttcacctcgc 3660aactagacca attcatcaag gacagggcag
gacaggatct ggtatctgaa cggaacaacc 3720gaaggaaaaa caaatcatac acaagctttt
gtacaatcaa accataagaa tcagtacagc 3780tttctctttc agcctttcca aatgagagga
aggacagaga aaatacaaag atgataccct 3840agaaacgaaa
385043139DNAAspergillus niger
4attgagtagg ctgcgctctt tgattctttg cgaggtacta aacccaaaca ggcaattgag
60atgttcggga gagaatatgc ggggatgcta tgcctgttgc agggatgatt atctccacac
120tagcgcgtta aaggtttagt agcctgagat ggattgccga tctccgaccg ggccctgcct
180cagccccagt acttgtggaa cgcgtcgaag atttacaaca ccatgccaga cctaactagg
240taataaagtt gcgatgatgg ggactgccgc aaggctgtca atttgggggc cgaactcagc
300ctcaacctgg aaaaattaac tccgattgtt ttgatttgat catgggagat tacagtctac
360gtgaaatgag cttataaaga acgttggagt accggctctc aacctgtctt gtacctgcaa
420gctttcctca attcccttct atctttctta aaatcctctc attagacatc atgacgaggc
480agacttctct cgttcccagg ctactaacgc tagcctcact agctgcactt tcacaagcag
540agctaggcaa gatccaatgg aaaggatctt gcaacttgac cacttatccg gcattgatct
600gtggaacact agacgtgcca tacgactaca cggagtcaaa ttccagcaag acactgactc
660tcgacatcgc caagtggcca gcgaccaaga aaccagtctc ggagcccatc atatttaact
720ttggaggacc tggtgtcaat tcgttcgagg gccttgggct ttatggagag gaatttcagg
780cgtaggttgc agttctgctt tgataccgca tcttatgctg actgacattg ttctctagta
840ttcttggagg tcacaatgat ttgatagctt ttaacaaccg gtttgtcgct cttctttctt
900gagctagaaa catctactaa cgttgtacct ttatatagag gcgttggaaa caccatcccg
960ttctcctgct acagcgatga cgccacccgt gaactcgtcg cccttcaagc tcctaacgac
1020ggcagagcgt ccagcacggc tttgggagaa atctgggccc agaacgcaaa catcgcacag
1080gcatgctatg ctacgaacaa tcaaactggt agtcttattg gaactagctt tgctgcaagg
1140gacatcatgc aggtcgctga tgcgctcagt ggaaaggata gtttggtcaa ctactggggt
1200aaatatcacg gctgaaccga gtttatactt ttgctgacaa tctacacttt aggattctca
1260tacggcacta caatcggtgc tgttctcgca gccatgttcc cggatcgaat ggggaatgtc
1320gcgcttgacg gagtggacaa ccccagagag gctctttatg gatagtgagt ggcccttgaa
1380gtttgcccga tctggtatga ttcagacagc taattccttc cgaaaagcaa cgcacaagcg
1440gttgtggacg tcgacaaagt tttcgaagga ttctgcacgg gctgcatggc cgcaccggac
1500ctctgcccta tcgccaagga gtacaccagc gccgccaact tggaagccgc aatttacctg
1560atgctggaaa acctcaagta caacccgatt gccattcccg aaaccggtgg aatcgtaact
1620tggagcgacg tcaagtcgac catttttgag gccatgtacc tgccaagctc ttggcccttg
1680acctctgagc ttctttacta cgtgcaaacc cgcaacacaa cgatccttgg caactctgaa
1740gtatacgaca ccatcaaatc ctacggtcaa tcggcttctt tgacttcggc ttccgatgag
1800gtcggcacgg ccattacatg ctccgacaag catcgatctg ccaccattaa agaggtcctc
1860ccgtacgtca aagccagaca ggctctgacc aagatcggaa gtgatggctc ggacggcgac
1920atgagatgcg cgcagtggaa tccgaagatg ttcgccaagg agcgctactc cggtgacttt
1980gaagtcaaga cagccaaccc ggtgttgatt ctgagcaaca cttacgatcc agcgactcct
2040cttcccgcag cgaagaacct gacagagacc tttgagggaa gtgtcttgct cgagcagaac
2100ggatacggtg tatgtttata cttccccttc tcatcatatc aaaagtgagc aagcagctaa
2160cctactgatg atagcatact accctgtcta tgccatctct ttgcactgcc aaggccgtcc
2220gggcttactt caccaatggc acattgcccg ctgacggaac gatctgccag gtggacgtgc
2280ctctgttcac gaacttgacc tacaaggatg tgtggccgaa gagtttccaa cggagcgttg
2340agtcgaggga tgatgcgact atcctcaagg ctttgatgtc ggtccgtgat aagatgtcgc
2400gacgcaggat gtggtaagcg catgctatag atggggatgt tgaaaaccca aagcgatgac
2460tggataatgt tcttcatata atttattctg tgccgattcc ggcatacgct cgtcatgtaa
2520tagagtttag ttaatgttgc aagggattgt atagtattta tttgtacacc aacagcgctt
2580catggagacc ggaacttccc tgagtacaga atttcccatc taggctttta tttaactttg
2640acaatactca gtagtttgga tgtcgatagc attgaggcaa gtaccggttg ttgttttggt
2700agtggagtga atcaggactg tcccacaaca atccgctgat ctgaatgttt gatgtttcga
2760tctgaggtat ctgagctgcg agcctccaaa tcggcaaccc catacttcag ggatattcca
2820gaacattact agccttggag gctttgaggt ttagctgcag ggcggtcaaa ctaattgatt
2880tctctcaatt gtcctaccgc tattgagtga tctttagtgg atgagtggaa cggagaattc
2940ccggtcggat aatgcctgca ttagtattgg ccaacagagt attccatcaa cgactcccgc
3000gctgacattt aatccatgca caccctgaat tacctcgagc ttattctcgg tcctacattt
3060gatcctctca tactttccct tatgtccttg cccacttgct ttacagattg aatccggatc
3120cttcagcact cggcagaca
313952940DNAAspergillus niger 5atagcagaac agaacatgta tcttgtcaac
gaattgattg atttcagcac ggaaatgttc 60aaggcatgga acaattgctt ctgacccgga
accgcggatt gcaaggatgg atatgtggat 120cagatggtgt gaagaagatc tgtctgttag
catacttcat aagttccaag gaggagtaaa 180tgtggagtca taagagtaat atcgaatatc
atagaatggg gttgtcttgc taacggtgaa 240tcggactccg tcggctctgc cagcgatcgg
caggtcacgc gtctgcgtaa cttggtactt 300atcttcccag tcacctaagc actgttactc
cactcttctc tctttgacga taagaagata 360gcagagactc ttgataaaaa gggttgggag
catgctttga atttcttcac tccgagagct 420ttctgtcccc atctggtctc ttgttgtccc
cgatgtacta ctctctctgg gttgctgcct 480tggtggccgc gctgcccgtc tcccgggccc
agtttgtggc tccgcccacg gatctcattc 540ccaccaaggg atatctcgac atccccgtcc
gctacaaaca ggtccccacc ggcatttgtg 600agactgatcc cagtgtcaag agcttctccg
gttacgtcga tgtcgctgag catgagcaca 660tcttcttctg gttcttcgag gcgcgcaacc
aagatcccac cgaggctccc ttgaccgtct 720ggatcaatgg aggcatgtct gaccccggtc
attatttcct tccaattgct aaccgttcgt 780aggtcctggt tcctcctcca tgatcggctt
gttccaagag cacggcccat gcggcattga 840cgccaatggc tccgtctaca acaaccccta
ctcctggaac aacgccagca acatgctcta 900catcgaccag cccgtgcaga ccggcttctc
ctacagcatt ccggttcccg gctatgtgga 960ttcttccaca gacaatgtta ttgctctgcc
ctcccccgcc tgccccgact atgcagcgga 1020tatgttctgt ggcacttact cctaccccaa
cgtgagcctt acggctaatt ccaccgacaa 1080cgccgccccc aacttctacc gcgccctaca
gggttttatg ggcgcatttc ctcagtactc 1140gcgcgaaacc ttccacttca ccacggagag
ttatggcggc cactacgggc ccgtcttcaa 1200cgagtacatc gaggagcaga acgcccatct
ccagccggga gccaagaaga tccaactggg 1260cagtgtgatg atcggcaatg gctggtatga
cccgattatt caataccagg cctactacaa 1320ctttacggta cactacctgg tttctgatcc
ttatccttac agcctggaca ctaatctatc 1380tacaggtata tccgggcaac acatacgact
acctgccatt caacaagtcc atcagctcgc 1440tgatgtacaa caacctctat ggccccggaa
actgcctcga ccagctctac gactgcgccg 1500cccgaggcat cgacgagatc tgcagcactg
ccgacgattt ttgcgccaac gaggtcgaaa 1560acgtctacga catttactcc ggtcgggatg
agtatgactt tcgtgaactc actccggacc 1620cgttccctta cgagttctac gttgactacc
tgaacaaagc gtccgtgcag gccgccatcg 1680gcgcatacat caattacacg gagagcaaca
acgctgttgg actcgccttt tcgtccaccg 1740gtgacgacgg gcgactcatg aacaccatcc
aggatgtggg caagctgctc aaacagggtg 1800tcacggtggt catgtacgcc ggggatgccg
actataactg caactggctg ggtggggaag 1860ccgtgtcgtt gcaggtcaag gccgccaact
tcagtagtgc gggttacacc aacattgtca 1920cctcggatgg agtgacacac ggccaggtgc
gccaggcggg gcaatttgcc tttgtgcgag 1980tgtatgagag tggacatgag gttcccttct
atcaaccctt gcttgcgctg gagatgtttg 2040agcgcgtcat tggcggcaag gatgtggcga
cgggaaagat tcccatctcg tcgagtttac 2100agacggtggg cacgcccaag agttactacc
gggagggcaa cagcacgatt cagtgggagg 2160tgttggattc tctggcgacg tacaacacaa
ccacgaatgc tccgaacccg gtgagccgga 2220ggctgaagcg gatgggacca gctttgcggt
ttcagatgta gatctgaagt acctgcgatt 2280gtgcattgaa gtacattgtt cagtgcaatt
accgatcatg gctagctgtg ccgcccaaag 2340ggggaagcta caatctaaac gccatatctt
tcgatgcaga caccttacga gccatgaaat 2400gccattttat gatcaggcta tctactgccg
gtgattttat gtagtgttgg aaattttctg 2460ttcaatcgca acccaaagga accaactggc
tgtaaaagac acaacataac tcgccagagc 2520caaactgtgc gaagatatga gaagcaaagt
ttaaacgctc ccaagccatg gtttccctgt 2580caagcctcat tcactctatt ggcttcagct
gaaagaagag gataaacatt caggggtaga 2640accagacaaa atctggtctg gcggggatct
ggtggggacc cgaatccaag ttgcgttagt 2700ttgaggcatg gatcgacacg cgacgaacat
gcagtggagg gccgctatcg ctgttttgac 2760tcgacatcgg accaaaacca aggaaattcg
tattcatcga tcgatagttc ttctcatctg 2820gcaaggcatg gatccgtctc cccacttcag
ccggtacctc ccggcacaac aaaaggagca 2880tggcagcctg atttcctgcg ttccaactgt
tcgtcggttg atctccctga tcaagtccag 294064550DNAAspergillus niger
6tgaaccatca ggtgggacca cagagcccac cgtccagctc aaggcagctc tctattcgaa
60gaaagaggta ctggatatat cgggtacttc tagctccagg tcgtgaagcg ttgcgagata
120cacaaatact gttggcgatc gaatgggacg aacatgtgaa tgttagcacc agaagtggat
180ggtagctacg aacgatggga ggcggaggtc cgagtgtacg atagagctgt ggagatagat
240tcgtatataa tagagatata tattcattgc attgcattgc tgtggtagaa aacttgtttt
300ggatcgcatg aattgctggc ctgaatgatc aggtcagaag acgccgcagc tccgccggac
360aagagccaat cagagggcgc caatcggggt gtccgcccgg ttcccttttc agtgtctgtc
420actgcagctc gcctgacttc agttcctttt ctccttcctc acggtcctcg tcattcgccc
480ctctctgttg cagtctctct tgttgggggg tcttgtgaat gtcttgcgtc tggctccaca
540tccacaaaag gagcctactg tctgtcgcta cgaacaattc tgttgcgagg gccgctgcct
600ctacctccgc cgcgccgccg ccgccgtcat cgccgccgcc tggttctaat acttattcgc
660ctctttatcg ccccatcacc aatcccatcg gatttacttt gtcgcctgcg aggtcactag
720tttctcgcaa tcctaaattt cctgcctatc ggcgctctag tcgacacttt tctttgtgcc
780cggccgctgc aacgcccggt gtcaccacga gcatctgccc tggtcaggcg cccgtccgct
840ctctcagctc gctcattata cactctacga gaccccgtgc tatacgtatc cgtaccgacc
900agatggattt gaatggagac gcaggcgcca agcgcaagcg cagctccatc accacacccg
960ccgaacggcc cgtaaagcac cttcgccccg aatcgagcgc attgacaccc ggggattcga
1020cgcctgccaa tgggactgta tacgatgtgg aggatgatga agatgcgagt cgtctgctgc
1080ctgtagggcc tgctcaggcc gactcaccgg aatggcaagc taccatagag gaggttgtga
1140aaagcgtagt gtctatccac ttctgtcaga cctgctcctt cgacacggag ctgtccatga
1200gtagtcaggc tactgggttt gtggtagatg cagagaatgg gtacatattg acaaaccgac
1260acgtggtttg cccgggacct ttctggggat actgcatctt tgataaccat gaggaagtat
1320gttgtacatt ccacatgtgg atcgccttat gacagtgatg ctgatttgaa ttggttacta
1380gtgcgacgtt cgtcctgtgt atcgggaccc tgttcacgac tttggaattt tgaaattcga
1440cccgaaggct attcgatata tgaaattgag ggaactgaaa ctgcagccgg atgcagctaa
1500agtgggatca gaaattcgcg ttgtgggtaa tgatgcagga gaaaaactga gtattctgtc
1560tggtgtcatt agtcggctgg atagaaacgc gcccgaatac ggcgatggct acagtgactt
1620caatacgaat tacatccagg ccgccgcagc agctagcggt ggaagttccg gcagtcctgt
1680agttaacatt gatggccatg cgattgctct gcaggccggt ggtcgtgcag acggtgcagc
1740gacggattac ttcctccctc tggaccgacc gctacgcgca ctggaatgca tccgtcgcgg
1800agagcctgtc acgcgtggaa cgattcagac gcagtggatc ttgaagccgt tcgacgagtg
1860tcgtcggttg ggcttgacgc ctgagtggga ggcgaccgtg cgtaaagcag cgcccacgga
1920aaccagcatg ctggtggccg agatcatcct gcctgaaggc ccggcggacg gaaagctcga
1980ggaaggagac gtgctcctgc aggtcaacgg ggtgcttctc acccaattca tccggttgga
2040tgacatcctg gattcgagtg ttgggcagac agtgcgtctg cttgtccaaa gaggcggtca
2100gaatgtggag attgagtgcc aggttggcga cctgcatgcc atcacgcccg accggttcgt
2160gacggtggct ggaggcacgt tccataacct gtcttaccag cagtcgcggc tgtatgccat
2220cgctactcgc ggtgtctacg tctgcgaggc tgccggctcc ttcaaactgg aaaacacact
2280gtcaggatgg atcatcgact cggtggacaa gcggcccact cgcaatctgg atgagttcgt
2340ggaggtgatg cgaacgattc ccgatcgttc gcgcgtggtc atctcgtatc ggcatattcg
2400cgatctccac acccgaggca ccagcatcgt ctatatcgat cgacactggc accccaagat
2460gcgactggct gtgcgcaacg acgacaccgg tctgtgggac ttttcggacc tcgcggaccc
2520tatcccagct cttcctccgg ttccgaggaa agccgatttc attcaactcg atggtgttag
2580ccagcctgct gcggccgaca ttgtgcgcag cttcgtacga gtatcctgta cgatgcccct
2640gaagctggac ggctaccccc aggccaagaa gactgggttc ggattggtcg tcgatgcaga
2700gaagggtttg gtggttgtgt cgcgagcgat cgtgccgtac gacctctgcg acatcaacgt
2760cacggtggcc gactccatca tcgtgaacgc taaagtagtt ttcctgcatc cgctccaaaa
2820ctacagcatc atccagtacg acccaagcct ggtgcaggcg ccggttcaga gtgccaaact
2880cgccaccgac tacatcaagc agggacagga cacgatcttt gtgggattca accagaactt
2940ccggattgtc gtggccaaga ccgccgtaac cgacatcacc actgtttcta ttccagccaa
3000cgcgtccgca ccgcgctacc gcgcgatcaa cctggacgcc atcactgtgg acaccggact
3060cagcgggcag tgttctaacg gtgtcctgat tggcgaggac ggagtggtgc aggcattgtg
3120gttgaactat cttggagaac gcacatctaa ttcgcataag gatgtggaat accatctagg
3180atttgcgact ccatctcttc ttcctgtcct gtcgaaggtg cagcagggag agatgccgga
3240attgcggatt ctgaacatgg agagctacgt ggtccagatg agtcaagctc gtatcatggg
3300cgtgtcggag gaatggatcg agaaggtgac gcaagctaac ccatcgcggc atcagctctt
3360catggtgcgc aaggtcgatt gcccaccgcc tgggttcaac tcagcggccg acacgttcga
3420ggagggtgat atcatcctga ccttggacgg acagctgatc acccgcgtct cggagttgga
3480tatcatgtac gagaaggata cgctggaagc cctgattgtt cgaaatggac aagaaatgcg
3540gatccaggtg ccgactgttc caacagagga cctagagact gaccgtgcgg tcgtgttctg
3600tggtgctgtg ttgcagaaac cacaccatgc ggtccgtcag cagatttcta agctacacag
3660cgaagtctac gtcagcgcaa gagtatgctc ctcacccctt aaccactgcc atcaagtaac
3720taaccaccct atcccttcaa cagagtcgcg gatccccctc ctaccaatac ggcttggccc
3780caaccaattt catcaccgcc gtaaacggcg ttccaacccc gaacctggac cgcttctccg
3840aagaagtgag caaaatcccc gacaacacat atttccgcct acgggcggtg acattcgaca
3900atgtgccgtg ggtagtgacc gtgaagaaga acgatcatta cgtatgccac aacccctccc
3960tctttacctc caccaaattc agagaatgtg gagactaaca caataatctc cagttcccca
4020tgtccgagta tatcaaagac cagtcccagc cttccggttg gcggaccgtg tctcacgaca
4080aggataaata taaagacggc attgcaccgg atgctgcgaa cttgaacccg gatgctatgg
4140acgaagggtt tgatggagtc agtgatattg agccggattt ggagtgattg aaggcggact
4200gtatagtatt ctgtgatgag catggctgtg ttagatgtgt atagcagaca tggcttggaa
4260tagtagaaaa agtgaaaaat gggaaaatga tagagattaa atcggttcgc atatacgaag
4320ttgtagatgg atgtattgtt tcggatgttt acgtagtacg tagtggctac tctatcgagg
4380tgtgatgata gcattagtct ttgatttgtt ttcgtcccat ctgcacatgt actccgtata
4440tgagaagtat gttgctcgac aacccaagtc tgaaagtatg aatgaacgaa tcgacaacac
4500acatgaagat aattttatcc aagcaaaaga agccccattt catcttattt
455072660DNAAspergillus niger 7gaatgcgctg aaaaaaatgc tgttgatagc
ctaaggtagt cacccgacgg gcttgtgatt 60ggaggaaacc gcaatgacgc tggtcccgtg
ccatgagctg actcagctgg ctgactcaac 120gcgtgacttt ctgcaaaggg aaggcaccgg
catgtgattg gtcgccttcc gttcctgcta 180tcgaggcgtc atcgcctaca gcccatcgag
tacacctcct cttctctcct ctctccttcc 240ttccttcctc tccatcctct tctgggtttc
cattgatcac ccagtagctg gtattttctg 300agctacttgc tttttctatt ttgatacttt
tgtgtccgta agttcacttt atgccctctg 360acccccccct tgataacccg tgctgataag
ctccctgccc aggtaccgta ccttccagac 420cgcaaggtac ccatcctctg cctactcatc
ccatcaccat ctcaattcat accggccccg 480tagggtttca gcaacaatga gagtccttcc
agctgctatg ctggttggag cggccacggc 540ggccgttcct cccttccagc aggtccttgg
aggtaacggt gccaagcacg gtgccgacca 600tgcggccgag gtccctgcgg atcacagtgc
cgacgggttc tccaagccgc tgcacgcatt 660ccaggaggaa ctgaagtctc tctctgacga
ggctcgtaag ctttgggatg aggtggccag 720cttcttcccg gagagcatgg atcagaaccc
tctcttttcc ctccccaaga agcacaaccg 780ccgtcccgac tcgcactggg accacatcgt
ccgcggctcc gacgttcaga gcgtctgggt 840cactggtgag aacggtgaga aggagcgcga
ggtcgatggc aagctggaag cctatgatct 900cagggtcaag aagaccgatc ctggctctct
tggcatcgac cccggcgtga agcagtacac 960cggttatctc gatgacaacg agaatgataa
gcatttgttc tactgtaagc acaccttggt 1020tcaagatcac gctttttata tgctctggat
atctaacgca acttagggtt cttcgagtct 1080cgcaatgacc ccgagaatga tcccgttgtt
ctgtggctga acggtggccc tgggtgctct 1140tccctcaccg gtctcttcat ggagcttggc
cctagcagca tcaacaagaa gatccagccg 1200gtctacaatg actacgcttg gaactccaac
gcgtccgtga tcttccttga ccagcctgtc 1260aatgtcggtt actcctacag taactctgct
gtcagcgaca cggtcgctgc tggcaaggac 1320gtctatgcct tgcttaccct cttcttcaaa
caattccccg agtatgctaa gcaggacttc 1380cacattgccg gtgaatctta tgctggtcac
tatatccccg tcttcgcttc ggagatcctg 1440tctcacaaga agcgcaacat caacctgcag
tccgttctca ttggcaacgg tctcaccgac 1500ggatacaccc agtacgagta ctaccgtccc
atggcctgcg gtgacggcgg ttacccagct 1560gtcttggacg agagctcctg ccagtccatg
gacaacgctc ttcctcgctg ccagtctatg 1620attgagtctt gctacagttc cgagagcgct
tgggtttgtg tcccggcctc catctactgt 1680aacaacgccc tccttgcccc ttaccagcgc
actgggcaga acgtctatga tgtccgtggt 1740aagtgcgagg atagctctaa cctttgctac
tcggctatgg gctacgtcag cgactacctg 1800aacaagcccg aagtcatcga ggctgttggc
gctgaggtca acggctacga ctcgtgcaac 1860tttgacatca accgcaactt cctcttccac
ggtgactgga tgaagcccta ccaccgcctc 1920gttccgggac tcctggagca gatccctgtc
ttgatctatg ccggtgatgc tgatttcatt 1980tgcaactggc tgggcaacaa ggcctggact
gaagccctgg agtggcccgg acaggctgaa 2040tatgcctccg ctgagctgga ggatctggtc
attgtcgaca atgagcacac gggcaagaag 2100attggccagg ttaagtccca tggcaacttc
accttcatgc gtctctatgg tggtggccac 2160atggtcccga tggaccagcc cgagtcgagt
ctcgagttct tcaaccgctg gttgggaggt 2220gaatggttct aaagacgtgc taccaccgca
tatagacttt ctggtcattt cggtgacact 2280gcagatatgt ttcttaacga tagtttgagc
atgcttgtca atgcccacta gtcccgatcc 2340ttatatgttg catggtatct atgagttttg
tcactatagt gcattataca tttgtacttc 2400gtatgagaat gaatcgatcg catttacacg
catataaata gtacccaaac cgtctggaca 2460tgaataaggc ccggccagta gtttacatac
agtgtagaaa actaggcgta cagacgtctc 2520agtacgtaat caatggttaa aaaaaccact
cccatagaag ccaagccata agagcctact 2580catgtagttc gccactgaac gcacccgtat
atcgtaaacc agcagaaaga gaaaaggaaa 2640attgaggaaa ggacgatttg
266081680DNAAspergillus niger
8aaaacgtgcg cactgcaccc actcgttccg gctggggtct agaaatctgg acggtcccag
60gcagatcggt gcctgggcaa aaccttgata aaaatagctt gttcgatctt gagttagaca
120gccaattgta tactcactag agacacttga tgattcagtc tgtgacgtac gtgcacctcc
180acactccgtc gatggattat gtgtccccgt gggcacgcgg agatcgggga catcagtcga
240gaaacttgcc taatctagtg acaagcagaa aagtcaaagt catccgcatg cgacttgcag
300atcatctaga gggatataat aaagtcgtgc gtttatgacc ttgcaggaaa cgacccgcct
360cctcctgcct ctttattatc gataccctct gccactagcg tcgtacatca cacttcacaa
420tccattctcc tctcattcat catgaagttc acaaattatc tcttgacgac tgcaacgctc
480gcaagcagtg tcctagcggc tcctgctccc cgcaccggtt tggaggacag actccgtgcc
540cggtcattgc agcgtcaatc acatcctctg gcacctattc cacttgacac atccaccaaa
600gagaattcca gactcctcga agccgacgag aataccaccc atgttacata cagcagtaac
660tgggcgggcg cagtgcgcga gcaaccacct ccgcaaggca cgtattctgc cgtgtcggca
720acctttcgtg taccagaacc cacggcgcaa ggggggagcg gaacgcaggc tgggtcggcc
780tgggtcggga tagatggcga cacatacagc aacgccattc tacagacagg agtcgacttc
840tacgtggaaa acgggcagac gtacaacgat gcctggtatg agtggtaccc agactatgca
900tatgacttcg acctagatgt aagcacaggg gacacgatcg tcgccaaggt ggaagccatc
960tcgccaagtc aaggtgtagc cactattgag aacatatcga cggggaagaa ggccacgcag
1020acgatcagag ccccagctgc gacagctacc cttgccggcc agaatgccga ctggatcgtg
1080gaggatttcc agtctggcga ctcaatggtc gatctggctg gctttggcga gatcagcttc
1140tggggcgtgc aagcacaagg aggagggtct acatggggtg tagatgatgc gactattgtc
1200gaactgaagc agggcaacga agtgttgaca gacgtggagg tgcaaagtga ttcggccttt
1260acggtgaaat atacgagctg atgtgatggt atatggtcgc atattctcat ccttcgacta
1320atctttctcc ttttggacgg tgacataatt tttctggttg attttgaaaa tgccttgatg
1380tatttcggta gtagttttaa ggttgatact tccagcatgc gggctcggct attgggagga
1440accaagaagg ctatatggcc ctcgggtatt cgccgcattc taaggtcgca tgccttctcg
1500gctttcctta tatatatcta ctttccattt ccccaacctc attcttgatg cgagaatcta
1560cccacacata cagttaatca tgtcccgtat ataccaacta agccgagcat ttcagcgcag
1620ccaagttggc gcgctccggc cgcgctctct actacccgct cggtcatccc gctgcggctt
168092590DNAAspergillus niger 9ttacatgctc tgtgtgttgg taaacacgct
gtcacattct tcatgtcctc tggattgcac 60gataacttta ccgcgtctct ccaaccggat
tgatgaaact atcaacaggg ccccgcatta 120ttggcataac cgacattttg aaaaagtatc
gcgggcgaga agagagcctt tttcgacaac 180aaacgtcggt ctctttggtt tgcatcgaga
gacatagata cagttgaccg cggcgcgaca 240ctttttagga gcgcagagga cggtttctgc
ggcgatttga tccgcaggcg agcaaaggtc 300taagacacct ctgtttggaa ctgcgcaggt
accagaactc gaaactaatg cgacattcga 360gcgacgatga tatatgtcaa ctatatcctg
ggacttctgt ccctcttaca caccgctgta 420gccacagctc ctgattatgt cgtggtagac
caactgaaca gcatccccga cggatggaca 480aaaggcgcag ctcccccgcc atttactccg
atgaagttct ggttgtcgat gcatcacgag 540tacaaggcgg acttcgagca gaaagtcatc
gatatctcga cacccggtca ccgggattat 600ggacggcata tgaaacgcaa cgatgtcatg
gcctttatgc gcccatccga tcaggtctca 660aagatcatct tctcttggct tgagtcggag
catgttccac caaatgccat cgaagatcgc 720ggggattggg tcgccttcac agtcccgttg
gcccaagcac aatcaatgat gaagaccgat 780ttttacaact tccaccacct ggaaacaaac
acaacccaaa ttaggaccct caagtactcc 840gttcccgagc aagtcgatgc tcatctgcaa
atgatccagc caacgactcg cttcggccga 900cctaagacac aaaccagcct accgagcctc
atgccagtgt cggttaacat tgatgaaata 960agcgaagact gcttgacagg cgtgacgccc
atttgccttc gccagctcta tggtttacct 1020agcaccaagg caagccccga ctcgagaaac
gtcctcggaa tttccggcta tctggaccag 1080tacgcgcgct acagtgacct cgacgagttt
ctagccgtat actctccaaa cagcgtagac 1140gccgacttct ccgtagtatc gatcaacgga
ggccaaaacc cacaaaactc acaagaggga 1200agcacagagg ccagtctcga catccaatac
gccctctcca tggcatttga cgctaacgcg 1260actttctaca ctaccgccgg acgtgcgcca
tccgtctcag actcgggtac ggtgagcacc 1320gacggctcga ccaacgagcc gtatctcgaa
cagctccagt atctggtggg tcttccggac 1380gaggatcttc ctgcagtgct tagcacgtct
tacggcgagg atgagcaaag tctgccggag 1440gaatacacag aagccacgtg caatttattt
gcccaattag gtgcacgcgg ggtctcggtg 1500atcttcagca gcggagactc gggcgtcgga
ggatcgtgtg tatctaacga cggaagccag 1560aggacccgct ttcagcctat cttcccggcg
tcgtgcccgt ttgttacatc cgtgggtggg 1620actgagggcg tcgggccgga aaaggctgtg
gacttttcga gtggagggtt ctccgagcgc 1680tttgctcgcc cgtcgtacca gaatgcgagt
gtggaagcat accttgcccg cttaggagat 1740aaatgggatg gattgtataa tccagacgga
cggggtattc ctgatgtgtc ggcccaggct 1800agcaactatg taatcaggga ccatgggcaa
tggctacaaa ctgcgggaac aaggttagtg 1860gccctaccaa gtcaacaatt gaaaatcaaa
cgagctgata atattacagt gctgccgccc 1920ctgtctttgc agcagtcatc tctcgactga
acgctgcacg tctcgagcag ggtaaaccta 1980cactagggtt tctgaatcct tggctgtact
cactcgacca gcaaggattt acggatattg 2040tagacggcgg atcagtgggt tgtgacgggt
caaatggagg agctcttgtc ccgtatgcca 2100gttggaatgc caccaaggga tgggatccgg
ttactgggct ggggacacct ctgtatcaga 2160ctctggagca gttggcgcag tctgcttagt
actgcggcgg gatggcctat tttgtgtcgt 2220tgatgttttt gtcccctaaa tgtcaacgcc
attaattctt ctcaagtgcg attagatttc 2280gtaaaacaga agctggacaa tatgcagaaa
ttagagtaca aagctggaga cagaaggtcg 2340atctccagaa tttgctaatg gttgtggtac
tcacgtcagg gtgctcgagc acaccagcta 2400actaccaact ttacccgtta attctaccgc
agtagtaatc tagaaaatac tagataagct 2460gatcaagctt gaaacaaata agcttaccga
ccgagaccca tacagcctcg cagtacaaca 2520acttactacg ggtgagacat cctcacccgt
ccgtaagatc gaaaggatta ctatacacgg 2580agtaagcact
2590103080DNAAspergillus niger
10ggtacagtag tgggttttag tttggatgta tctggagaca tcgatgcttg ttcgccggcg
60ctaatttggg gccatcgctt agggaagaaa aagaaaggaa ggatttcatc tgccgcggca
120tttttggtca ggcattggca atactccaag cagcttgcca aagtgagaaa tgtgcttatg
180aataatattg agtaatagtt aatcaattgt cttacaggat aatatgggaa tagcattggc
240attgcgtggt tggcaggcag catgttgcaa attgcaacaa cgcgatcccg gcgatcgccg
300tcggactccg ggtgggaatc agggttggag acccattgct tatcaacaac agccgacgcc
360agtcgttctg gggtgcattt gcagaatctt tggacgaatc agccttttgt cagtatcggc
420cggtccgaat tgacccagct tgctagccgt tggagacatg caatgatgcc attgtctcgt
480ccgataagca agtgcggcca atccctcgtt ccttctggct gtctcagaaa agttataatg
540tccgtctcgc ctgcctagcg aacaattcga actctgtgtg tgttgtccct ctcaaggtct
600ccagcatgcg ttcttccggt ctctacacag cactcctgtg ctccctggcc gcctcgacca
660acgcgattgt ccatgaaaag ctcgccgcgg tcccctccgg ctggcatcat gtcgaagatg
720ctggctccga ccaccagata agcttgtcga tcgcgctggc acgcaagaac ctcgatcagc
780ttgaatccaa gctgaaagac ttgtcaacac ctggcgaatc gcaatacggc cagtggctgg
840accaggagga tgtcgacacg ctgttcccgg tggccagcga caaggctgtg attaactggc
900tgcgcagcgc caacatcacc catatttccc gccagggcag cttggtgaac tttgcgacca
960cggtcgataa ggtgaacaag cttctcaacg ccacctttgc ctactaccaa agcggctctt
1020cccagagatt gcgcacaaca gagtactcca tcccggatga tctggtcgac tcaatcgacc
1080tcatctcccc aacgaccttc ttcggcaagg aaaagaccac tgctggtctg aaccagcggg
1140cgcaaaagat tgacacccat gtggccaaac gctccaacag ctcgtcctgt gccgatgtca
1200tcacgctgtc ctgcctgaag gagatgtaca attttggcaa ctacactccc agcgcctcgt
1260cgggcagcaa gctgggcttc ggcagcttcc tgaacgaatc cgcctcgtat tctgaccttg
1320ccaagttcga gaagctgttt aacctgccct cccagagctt ttccgtggag ttggtcaacg
1380gcggtgtcaa tgatcagaat caatcgacgg cttccttgac cgaggcggac ctcgatgtgg
1440aattgctcgt cggagttgct catcccctcc cggtgactga gttcatcact tctggcgaac
1500cgtgagtatt gaattcctag acagagcttg aatcgaagct aattgtgtag tcctttcatt
1560cccgaccccg atgagccgag tgccgccgac aacgagaacg agccttacct ccagtactat
1620gagtaccttc tctccaagcc caactcggct ctgccccaag tgatttccaa ctcctatggt
1680gacgacgaac aggtacgacc ccacattccc ccttccctgt atgtagatac taaccggacc
1740agaccgttcc agagtactac gccaagcgag tctgcaacct gatcggactt gttggcctgc
1800gcggcatcag tgtcctcgag tcgtccggtg acgaaggtac acttacttcc cctgtcctat
1860tccttgcatc gaaaataccc taacagcagc acaggtatcg gatctggctg ccgaaccacc
1920gacggcacca accgaaccca attcaacccc atcttcccgg ccacctgtcc ctacgtgact
1980gccgtgggag gaacaatgtc ctatgccccc gaaatcgcct gggaagccag ttccggcgga
2040ttcagcaact acttcgagcg ggcgtggttc cagaaggaag ctgtgcagaa ctacctggcg
2100caccacatca ccaacgagac caagcagtac tactcgcaat tcgccaactt tagcggtcgc
2160ggatttcctg acgttgctgc ccatagcttt gagccttcgt gagtccattc ccagcatcat
2220gataatacga aatagggtct aatgacatct gcagatatga ggttatcttc tacggcgccc
2280gctacggctc cggcggtacc tcagccgcgt gtcccctttt ctctgcgcta gtgggcatgc
2340tgaacgatgc tcgtctgcgg gcgggcaagt ccacgctggg tttcttgaac cccctgctct
2400atagcaaggg gtacagagcg ttgactgatg tgacgggggg ccagtcgatc ggatgcaatg
2460gcattgatcc gcagaatgat gagactgttg ccggcgcggg cattatcccg tgggcgcact
2520ggaatgccac ggtcggatgg gatccggtga ctggattggg acttcctgac tttgagaagt
2580tgaggcagtt ggtgctgtcg ttgtagatgt atactatata tatggtatga gattatgtat
2640gtgatatgtg atattatgtg agagagaatg gtttagactg tgcgtcatat acatggacag
2700ttcattttct cattaaacga gcaccttcat acggtaagga cctcagaggt tcctcccatt
2760gttatgaccg cttcccttct ttctagagat acatgcttcc ccaccccgcc tcaacgcgac
2820cgtctacgga caccagtcag aagaccccaa caacccacta ttagtggcta gtaaggacga
2880catgcataat tactccgtga aacccccgaa attaagcccg acaggggcat agagaaaccg
2940atccaatcac ttaacttacc ccgcttctag cacggcatag ggctgagcga tgcgatggtg
3000caatggccac gccgttgccg attagccgta gggccagttg tgaatttccg ggatcaccct
3060acaacacatg cttcatgcgt
3080111890DNAAspergillus niger 11gaggatgata gtctgatggc cattccgctt
gaggcaaagt gtgccccaat gtcacgcggt 60cagctagtct tctgcaggtc tgcagttgga
caggggaact tgagtgctat tatcgccaag 120gtgaatgttc tctaccgctt tgataccaga
gtgatcgtcc cttttgccgt cactcttggt 180gttgcgaagc agatgtcgtt cgtttgcata
tgggagccga agatccacgc agggtgcttg 240aaaagacata ctgcgatcct cccgtttcca
agaatgcttg tacacttgaa gtttttgtca 300agatcccccc gcctggccag tcccgagcta
agcgaaatga gctttccctc aatgctgcag 360acaggagaca ccatcgcttt ggcgccttgg
caccatcagc caccgtcggt cctatcatca 420tcgccctggt caatagggaa cgcgtcaatt
gcgtatctgg gtagtgttat ctgctcgata 480ttgcaccggc agagcaggaa tcgggattgg
cggatgtaga gaaaattcag cttcaaggag 540gggggaaggg agacgctgaa atggtataaa
accacgtcca attccctacc agatagccct 600catccagcag catcaaaagc atcttccact
cagactccaa gcagctccca gtccctcttc 660aattcattac cttccaaaca tcatcccatc
aagatgaaga ctactgctct cttgaccgcc 720ggcctgctgg ccaccactgc tatggccgct
cctctgacgg ccaagcgcca ggctgctcgg 780gccaagcgct ccacgaaccg ccagagcaac
cctcccttca agcctggcac caacgaggtc 840ctcgccctta acggcaccaa gaatgtggag
tacagctcca actgggccgg tgccgtcctc 900attggcactg gttacactgc cgtgaccgcc
gagttcgtcg tgcccacccc ctccgtgccc 960tccggtggct cgagccgcga ggagtactgt
gcctccgcct gggtgggcat tgacggtgac 1020acctgtgaca ctgctatcct ccagaccggt
gtggactttt gtgtccaggg cagcgaggtg 1080agcttcgatg cctggtacga gtggtacccc
gactacgcct acgacttcag cggcatctcc 1140atctcggccg gtgataccat caaggtcacc
gtcgatgcca gcagcgacac caccggtact 1200gccacgattg agaacgtgag cactggtacc
acggtcaccc acagcttcac gggcggtgtt 1260gatggtgatc tgtgtgagta caacgctgag
tggatcgtcg aggacttcga ggaggatgac 1320tccctcgttc cctttgccga ctttggcacc
gtgactttca ccagctgctc cgctaccaag 1380gatggttcct ctgttggccc tgaggatgct
accatcatcg acatcgagca gaatgaggtg 1440ctgacctccg tttccgtctc cagtagcgag
gtcgttgtca agtacgtcta agacgttgga 1500tgtatggggt tcgggattgt tgatgccccg
atgggtgtgc tttcgacggc aatggtgaga 1560tgagtgatgg aaatgagagg ttggtctttt
ggcccggggc tagttttctc ttggtactcc 1620tgaaaagcca atagttcaaa tgtctttatc
ttgttagttt gatgttatag tttgccttga 1680gttgatttaa ttgaaccaac gttggatcat
cgtatccctt agcgcaaata aaatattacg 1740tacgcctttt aaaatattat gtatgtctgt
taaggttctg actgttcaga tttatgtcaa 1800acaacagcaa atcattggta atccaggact
cggggatatg agcttcagac attcacccca 1860ttactgcatg actatgaccg tgtctttagc
1890123080DNAAspergillus niger
12ttgttgtgat tgctaagaga catcacattg ctcatatgag tttgattgcg actgttaata
60ttaatgtccg actgggagga aatgagtggc gttatccagg cagctgcatg caatactaca
120ttcccctcac gagggaaatc tggccatctc ggaaatcaat ggtgtcatcc gagttgattc
180ccctcaataa tggtcctgat aagccactat tgctgcagaa cgcctatctg tgccgttagt
240gcatgcatag tgtccccagt tctttttctg ccgattaatc tattgccaga cggactccgt
300cgcgcttccg aatgccggga accgcggcta agctttgctg tagctttgct gcaggaacat
360ctgcatcttt caaaatatct tcagaggagc cgatcgctga gtttctcaag agattagatg
420aatgttcgta cttggccgca atataagctt catggcggct tcaaccttcc atgagaaata
480agaataaggc tgccggggat ggcctcaccg tgtccctgca tgcaagggat gcataatgca
540ccgtgacctg ttccaccacg ccggagcccg cccgccctcc gacggcccaa acccactgca
600gaatggcttg attaagttcc ccttctataa ctaaccggtt gttcctcgtc gttgtctgct
660ctgctctcca cccagggtcg tttttgcttt tgcctcctta accatggtcg ccttttcccg
720catctcggca ggcttcgccc ttgccgcccc tgccctggcc agcgtcgtcc tggagaccgt
780caagtctgtt cccagcgact ggaagctcgt ggaggctgct gataccagct ccacaatttc
840tttgtccgtt gctctggcgc gtcagaacct ggaccagttg gaggagaagc tcctggccgt
900gtccacccct ggcaaggaca cctacggcca gttcttggat ctggacgaca tcaatgagca
960gtttcctctc gcagatgacg ctgctgttgt ggcttggctg aagaaggcag gcgtcaccca
1020gatccataag gagggtggtc tgctgaactt tgcgaccact gtgggcacag ccaaccagct
1080tctcaacacc accttctcgg tgtacaagag cggatctacc cagaagctgc gcacaacgca
1140atactctgtt ccggatgagc tgaccgggtc cattgatctc atctcgccga ctgttttctt
1200tggaaagtcc aacgctgcgc gctcggcggc cgtgcgtgct tcgcagacta ccaaggagac
1260cagcagaaag aagagcagta atgtgtgcga gtacatcact ccggattgcc tcaaagagca
1320gtatagcatt gactatacgc ccgaggcatc gtcgggaagt cgtgttgggt ttggcagttt
1380cttgaacgag tcggccttgt actcggattt ggatctgttc acccagtact ttgacattcc
1440ccagcagagt ttcactgttg agactatcaa cgggggaatc aacaaccagg agaatgatcc
1500ggatggtgaa gccgatctcg atgtccagaa catcgtgggc atctcgcatc ccttgccggt
1560gacggagtac attaccggag gatctccgtg agtgttccca aagatgcaat tgaattaaag
1620ctaatggttc agtccattca ttcccgacgt cgagactact accgacgaga acgagcctta
1680cctgcagtac tacgagtatc tgctggccaa gaccaacgac gagctgccac tggttatcag
1740caactcgtac ggcgatgacg aagatgtaag catccctgcc tcccacacaa atcgcctgct
1800gacagaatag accgttccca ttgcctacgc cacccgcgta tgcaacctca tcggcctgat
1860gggcacacgt ggtatctcca tcctcgagtc ttccggcgac tctggtacgt tgtaccccat
1920atatattgca tcaagtcccg actgacaaat acaggtgtgg gcggcgcatg catgtccaac
1980gacggcaccg acaagaccga attcaccccc atgttcccag gaacatgccc gtacatcacc
2040gcggtcggcg gcacccaaga cgtgcccgaa gtcgcctggg tggacagctc cggcggcttc
2100agcaactact tctcgcagcc gtcgtaccag tcggatcagg tggagaccta cctggacaag
2160tacatctctg cctcgacgaa gaagtactac gagcagtaca ccaacttcag cggtcgcgcg
2220ttccctgacg tgtctgcgtt tgcaggttct ccttagtatg tatccatccc agatgattgt
2280atggacatct gctaatgtcc gacagctacg aaacttatat tgatggtcag ctcggccttg
2340tggcgggtac ttctggcgct agccctgtgt ttgcggggat cgtcgcgctg ctgaacgatg
2400cccgtctgcg ggccaacaag acatccttgg gcttcctgaa cccttggctg tactcgagcg
2460gctacaagag cctgaatgac attaccagtg gcgaggcagt gggctgccaa ggcgatgtgg
2520agggcgctgg agtcattcct tgggcgagct ggaatgccac gacgggatgg gatccggcga
2580cagggctggg aacgcctaat tttgccaagc tgaaggaggc ggttcttgcg ttgtaagcag
2640gaggatactg agtggacgtg cggagtgaag gatatgtgca aggcgttaac ttataatagc
2700ctgtgtgtgg actatagaat catctacagc ccacaaccaa ttagcttctg catagtcacc
2760acccttaaac taaggataat caattgttta ccgctgtcag tctcaaatcc gtcatgtcat
2820ggaccctgtt cgatcaactt gaaacaagct taatctacca gcggcggtga tgaaaccgcc
2880agcggacact tgtttcagtt acccgcaagc ctgtatccag tatgcaagcc ctacttaaac
2940ccttcaccac cctgattatt ctactctctc tctctctcca accacaccat tcttttcttc
3000attgtgccgg tcacatccat tctttacttt cattctcctc ccttctcttc tcttctctac
3060ttccccaagt cactcattat
3080133598DNAAspergillus niger 13accggagcag aggaacaagc ggagaagctg
aaacgtatga atgctgagta cgagcaaaag 60ttccctggct tgcggtttgt gtatgttctc
tcagtatacc caacgacttg tgttggtctg 120acatatctcc ctggcatagt acttttgtaa
acggtcgcaa ccgggatgtc atcatggagg 180agatgcgtga gaggattgac cggggcaatg
ccgatcgtga ggtggaggag attatacagg 240tgagatggct gtctacccaa acacggagca
gtgctgacag ttcgctcctt taaaggcaat 300gtgcgatatt gcaaaggatc gggcgcggaa
actggagcag tccgctaaga tatagttcac 360atactgcaag catatattta aacaatgaat
acccttcgat ttcagcaggc gctgcgacat 420ctccttgctg aatcgctgct ttacaagcca
tatttcggtc catgcgattc atctatggag 480cactctgcct ctgtccggaa acaatggctg
gtaatctgca tggaacttgg tgattccgtt 540agtataaagt gaccaactag tgccggcgaa
ttatgggaag cctgccttgt cttacgacgg 600gacgatcttc cgtcgaacgc gtgctaccag
cattttcata gtatgacggg tccgtatttg 660atttactatc gatggtaaaa cgcctcctct
atggggtccc ctagcactcg gcaggccagc 720ttgtgagaca acccgggagg tcagccatta
ttaggccaat gagagctggc aggttggaga 780ggttgtactt cgacaatgtt ggacagccat
gcaacgtcgg acaatagctt tgatagggaa 840tatcctaagg gcgaaccagg agggataggg
gtaggtatgg ctgtatcgat tagtagtccc 900tctttcaccc agaacaaact ctagtatata
tagtagtaaa tgttccgcgg atggccaatg 960accccaaatc atctttcatc ccatccacgc
atgagcaaac atgcatggtc tgcgcctagt 1020atgcagcata gggacattgc ctttggttat
cctggcatat ccggcggctt cattgcatac 1080aacttcagca gccgtggact tggactccct
tcgtctgacc tctaactccg aatacgtcaa 1140ttctgtccat gtagacacga atcgatcagt
cgcagtgtcc gctgaagaac attataccga 1200tacagcagct cgactggttc agaacattgt
tcctggagcg agctttcgtc tcatcgatga 1260ccactttgtc ggcgacaatg gagttgcaca
tgtatacttc cgccaaacgc tccatggtat 1320tgacattgac aatgcggatt tcaatgttaa
tgtctgctcc ccaattactc tgttcggaag 1380gaagcttact gttacagatt ggaaaagatg
gactggtctt gtctttcgga cattcgttct 1440tcacaggcgc gttgccgagc agccatctgg
acaataccaa cgttttgagt ccggaggctg 1500cacttagagg agcaagggac gctatacagc
ttccactgac tattgacaat gtttctactg 1560aagctgcaga ggggcggaac gagtacatat
tcagagaggc agtgggagcg gtatctgacc 1620ccaaagctaa gctagtctac cttgtcaagc
cagaagggac tctggcgctc acctggagga 1680tagaaacaga catgtatgag cactggctac
tgacatacat tgatgcagag actaccactg 1740tccacggcgt ggttgactat gtcgcagacg
cgacatatca agtttagtga gtggctctcc 1800ccatcgaata acgaagctaa gacaacagtc
cctggggcac aaacgatcca gcagaaggac 1860atcgcaccat tgtcaccgac ccctgggacc
tatccgcatc cgcatacacc tggataagcg 1920atggacggga caactacacc acaaccagag
gcaacaatgc catcgcacac tggaatccga 1980ccggcggtgg ctcctatctc tacaacctac
gtccatccga ccccaacttg aatttccaat 2040ggccatactc cccaaacatg tccccacccc
gatcatacat caacgcctcc atcgtccaac 2100tcttctacac agcaaacgcc taccacgacc
tcctctatac actcggcttc accgaatccg 2160ctggcaactt ccaatggaat aacagcgccc
acggcggccg agacaaagac tacgtgatcc 2220tcaacgcaca agacggctcc gggttcagca
acgcaaactt tgcaacccca cccgatggta 2280tccccggccg tatgcgcatg tacatctgga
tcgagtctac tccgtcgcgt gatggaagtt 2340ttgacgcggg cattgtaatt cacgaataca
ctcacggtgg taagcatctc cccagaagat 2400ggaagtccta atctaacaaa ccagtatcca
atcgtctcac cggcggctcc cacaacgccg 2460gatgcctcag cgccctcgaa tcgggtggca
tgggcgaagg ctggggcgac tttatggcga 2520cggccatccg aatcaagccc aacgatacac
gcacaacgtc ttacactatg ggtgcatggg 2580cagataatga taaatgtggt gtccgggact
atccttattc tacctccttt actgagaacc 2640ctttgaacta tacgagcgtg aataccatga
acggcgtgca cgccatcgga actgtctggg 2700caaccatgct atacgaggtc ttgtggaacc
tcatcgacaa gtacgggaag aatgatgggt 2760cgaggccggt gtttagaaac ggggtgccta
cagatggaaa gtacttgatg atgaagttgg 2820tggtggatgg gatggcactg taagtgatgc
acagaatagc ttctttcacc tttgtaatgt 2880ccgtggtatc ctgactaatg gactgaaggc
aaccatgtaa tccgaacttc gtgcaagcca 2940gggacgcgat ccttgacgca gacattgtgt
tgactggcgg gaagaatcgc tgtgagatct 3000ggagggggtt tgcgaagaga ggattggggc
aaggagcggc tcatagtagt ttaaattgga 3060tgcggagggg gagtacactt cttcctacgg
gatgttagtc ttgcacaggt cgttcttggt 3120gagagatgag tggagatgcc tggtttatta
gaataggact aattatctag cactaataca 3180atatcgacag atctaaacag taagtataaa
tctataagtt atcacacgca taaaccccat 3240caccccctag tctacttcga aacaaacacc
acaccaccaa catcctcatc ctcaccaaac 3300acccctcccc cgcataagaa cctaaacaac
agccctcaac tccatactct gcgcccccct 3360cgccacacac aacaacaacc cacccacgag
cagactcacc cccgcgtata tcgacagcgc 3420cagaaaccca tacttctgca ccagcgcccc
ggaaataggt atcccagcca tagcactaca 3480ctcccagata agatatgatt agcaaaatcc
acccttactc cccaagaaga tcaattcaaa 3540ataaatctaa tgtgcatacc tcaaagccgt
agcccccata acaaccccca ccgccaac 3598142847DNAAspergillus niger
14cttttggctt gtgatcttga ttgctagaga tgtatatcct cacggatacc gccggagtgc
60gccatttctg gttaccttct ctttcccttt ttgtctcgat cgtgaggcgg aacgcaggat
120gaagacacgg cttctccatc gcggcccacc aaccaacaat gtccttggac gcccaactct
180ccatctactg gtcattggtc caatgcagag actccgtcga gctcaaatgg gccggccaac
240cccgagtcgt caggggcagc ggcagcgacg agctaaatta gaccactgat aagacgcgat
300agtccaaagt ctgaccgtca cattgtgcca ggcagataag ttgaatcgtg tgactggatg
360ttggctaacg tatggcgtct ccggaggccc gacggaccct gcgcgatcgg cggtggagcg
420caatctaagg acatccgcgc ctaagatatc tacccttcag cagttcagcc tagccctgca
480gacttgtcgg accagtgcta tcgtgatcgg cccccacggt cgaatgagct cttgtctctt
540tccgtcagac cctgccagtt aatctgctat ctactccgcg gtaacatcgt gcctgtctcc
600actaaggcag ggtccagggc tgtatgtctt actttgcacc gagtcggccg ccggttggct
660ctgtcttggc aattgcgaat atcctcacgg gcgacggacg acacggattt ggacggacat
720gcggagatct tcgtcggttt attcctggaa gggacatcat ctccttccat catgacggct
780gccatagcgg ggactctgag acatttttgc tctgaagagc atggtcgact tggatgatgg
840aggagttgat cgaggtcaat gaggagaggc ttgcaagtat aagaagagac tgctcgacca
900gcagaatgga tcttcttgtt catcaaccaa gagtccaagg cttctttgtc tggttctatc
960tcttctccga actctcttgc ttgacattct cgtggtcaaa atggtcgtct tcagcaaaac
1020cgctgccctc gttctgggtc tgtcctccgc cgtctctgcg gcgccggctc ctactcgcaa
1080gggcttcacc atcaaccaga ttgcccggcc tgccaacaag acccgcacca tcaacctgcc
1140aggcatgtac gcccgttccc tggccaagtt tggcggtacg gtgccccaga gcgtgaagga
1200ggctgccagc aagggtagtg ccgtgaccac gccccagaac aatgacgagg agtacctgac
1260tcccgtcact gtcggaaagt ccaccctcca tctggacttt gacaccggat ctgcagatct
1320gtaagcttcc ctgctcgggt gttcgggcaa atcgtgacta acctggacta gctgggtctt
1380ctcggacgag ctcccttcct cggagcagac cggtcacgat ctgtacacgc ctagctccag
1440cgcgaccaag ctgagcggct acacttggga catctcctac ggtgacggca gctcggccag
1500cggagacgtg taccgggata ctgtcactgt cggcggtgtc accaccaaca agcaggctgt
1560tgaagcagcc agcaagatca gctccgagtt cgttcagaac acggccaatg acggcctttt
1620gggactggcc tttagctcca tcaacactgg tgagtcaatc ctacatcagc cgggttgacc
1680tacctgctga ccgatacaca gtccagccca aggcgcagac caccttcttc gacaccgtca
1740agtcccagct ggactctccc cttttcgccg tgcagctgaa gcacgacgcc cccggtgttt
1800acgactttgg ctacatcgat gactccaagt acaccggttc tatcacctac acggatgccg
1860atagctccca gggttactgg ggcttcagca ccgacggcta cagtatcggt gacggcagct
1920ccagctccag cggcttcagc gccattgctg gtaagaaccg ccttcattta acacacaact
1980tgtccacctc tttactaact agtgtataga caccggtacc accctcatcc tcctcgatga
2040cgaaatcgtc tccgcctact acgagcaggt ttctggcgct caggagagcg aggaagccgg
2100tggctacgtt ttctcttgct cgaccaaccc ccctgacttc actgtcgtga ttggcgacta
2160caaggccgtt gttccgggca agtacatcaa ctacgctccc atctcgactg gcagctccac
2220ctgctttggc ggtatccaga gcaacagcgg tctgggactg tccatcctgg gtgatgtttt
2280cttgaagagc cagtacgtgg tcttcaactc tgagggccct aagctgggat tcgccgctca
2340ggcttagatt atccactgaa gtggagtcta tgatctgctg attgatccct cgacgatgaa
2400ctacatgtgg aaatgcatag cagacgaggg tgatggtgat gatgttgatt tgatgatgac
2460ccgtacatac ttgatgaagc tcggtacata tgcaaatgtg actgtatcta tgtgatgaat
2520atatgtatcc atctcatggc ttttggctat gagtgcagga taaacacctg aaccagtagt
2580agtactttcc cacctatatc tactgcggtg cctcgtccgg cccaacatca ccccagaggt
2640ggccgcagag gagtcttata agatagctac tatcagttac aacacctctc tgacagatgt
2700gaaggagtac aataaatcac cgaaacacaa attcaactaa agtcggtaag taataataat
2760ttaagaccca atccacgcaa tgttaaacta tctctggtgt tgaaagatct ctcccctggc
2820aacacctagt tgtgggagaa ctgtgtt
2847152899DNAAspergillus niger 15gcccaggtga agaccggaca ggaagcggat
aggtacggga cattttctaa tctacccgcg 60atcgggacat ggctaaccaa gcatatagac
tcgaattcta ccggtaaatc aagtatggga 120cgtgcatcag gctggatatc ggattacgca
aggcgaacag ggggaccgtt agctgtatta 180tcaacatcta ggctatttca tattaggaca
acgactgacg cattgggtat tccgctgggg 240tagtcttatc ggttggggcc aagtaccttg
tagaactgta acccacgtta ataccgccac 300ttggctgggg cggtgattta gcatatgtaa
gctccagttg gacggctacc cgagcttccc 360atgatctaca ggagtacgtg tctggctgtc
tgctgcctac ttggtagaca ggtcagcgat 420aggtagatag gacctgtccg cagctgttgg
ctagtttggt aaggcggttg cgctagtttg 480aagtaggcag gcaccgggaa cctaaggcgg
tcttacatca tcacccgcgc tcggattcgc 540gtgatccgac catcacgata aggcctcagg
tagcaaggag accttccaga cagctctgaa 600tgagactcaa aggtagatat aatgatggaa
agataggata gctagatcag gcttattgta 660cctgatcgtt aagagcctag agaagatgta
cctggaagac ctggcagcta caatcacctg 720gagcgataac ccgtgacgat ccccttgcca
aatgacgcag ccgggctggc caaccattgg 780ctgcgacctg gcaggtcgct ccgcaaccag
cgccgcccgg ctccaagtca cccgcatcac 840tcttccctac ccccagacct cctcttttcc
cttgctatcc tccatctctt cttcatcgtt 900ctttgtctct atcatcattt tctattcata
cgtgcatcat tcagtcgttt ggcccagtcc 960atcatatccc gctgggtagc cgtttccgcc
gtcgcccatc atgaagtcag cctccttgct 1020cacagcatcc gtgctgttgg gctgtgcctc
cgccgaggtt cacaagctca agcttaacaa 1080ggtgcctctg gaagagcagc ttgtgagtgt
ggtctttcac tgctttgttt tttttagcta 1140gttagcttca aagaagctcc agaaccattc
aaagctgatt tcgtggtcta tagtacacgc 1200ataacatcga cgcccatgtc cgcgctctgg
gccagaagta catgggtatc cgcccgtcca 1260tccacaaaga gctggtcgag gagaacccta
tcaatgacat gagccgtcat gatgttctgg 1320tggacaactt cctgaacgca cagtgtatgg
agataccgtc ttcttatggc tgcaactgct 1380gacccttcct gccatagact tctctgagat
cgagctgggt actccccccc agaagttcaa 1440ggttgtcctg gacactggca gctcgaacct
ttgggttcct tcgagcgaat gcagctctat 1500cgcctgctac ctccacaaca agtatgattc
gtctgcctcc agtacgtatc acaagaatgg 1560cagtgaattc gccatcaagt acggctctgg
cagccttagc ggattcattt ctcaggacac 1620cctgaagatt ggcgacctga aggtcaaggg
acaggacttc gctgaggcga ccaatgagcc 1680tggccttgcc tttgccttcg gccggttcga
tggcattctc ggcttgggtt atgacaccat 1740ctccgtgaac aagattgttc ctcccttcta
caacatgctt gaccagggac tcctcgacga 1800gccggtcttt gccttctacc ttggagatac
caacaaggag ggtgacgagt ccgtggcgac 1860cttcggtggt gtcgacaagg accactacac
cggcgagctg atcaagattc ccctccgtcg 1920caaggcttac tgggaggttg agcttgacgc
cattgctctt ggcgatgatg ttgctgagat 1980ggagaacacc ggtgtcattc tggacactgg
tacctccctg attgctctgc ctgctgacct 2040ggctgagatg atgtaagtcg aattcttcgg
attcctgggt tgaaaagaaa tgctgctaac 2100aaccttctag caatgctcag atcggtgcta
agaagggctg gaccggccag tacaccgttg 2160actgcgacaa gcgctcgtcc ctgcccgatg
ttactttcac ccttgccggc cacaacttca 2220ccatctcctc gtatgactac accttggagg
tgcagggctc ttgcgtcagt gccttcatgg 2280gcatggactt ccctgagccg gttggtccct
tggccatttt gggcgatgcg ttcctgcgca 2340agtggtacag cgtgtatgac ctgggcaaca
gcgctgttgg tctggccaag gccaagtaaa 2400ttagttctgc gggttgatgt ggtatctatg
atgcagctgk tgctgtcatt attgcttctt 2460gtagcttgat ctatgatttt tgcagacgaa
cacacgtgat gttgtgaatg gttttcctca 2520tgtttgcagc ggttgccgga tagattctag
ggatcttcaa tggaaagccg gtgatattat 2580ttgaccttta tttgggcact gagaatcttg
actgtatgaa atatgatagt aacaccttaa 2640acatgaatgc aaatggcgta aaccgtgtga
tgcagtcaca ataaccagca accgcggtac 2700cagccgaagt ctgggccggc gagtctcgcc
ccgcacaggc caggcgccaa acccaagcag 2760cgctcttggc agccaagcct tcttcatcac
gctctcacca cctcctccat ccaggctttc 2820tctctgctcg gtctttgttt catcatcctc
ccacgcctgc ccttctattc aaccgctctt 2880cgatcttcat atccgatcg
2899162738DNAAspergillus niger
16gaggcatgag gcatttctga ggcccgctac tccgcattct gcagcatatc gtctctgcgt
60aggggaggtc gaaaccagct gtaggactcg gcttcggtgt atctgtaccg actgactaga
120aatcgctcaa tcgtgtagta tagctgtctc tttgttcctc acaacatgtc tacgatatgc
180tattaaaaaa agcagaagat ggagtcagag ccacccggtt agggccgggc cgcccgggag
240gagaacaaaa tacgggacag aatctcagtg atgggggaga agagagagtg gcgacctgac
300aattcacaca cgacacgaat aatagccgaa actaacaaga taaatcacat cacatcatga
360agaagacctg cgtaatgatg ataagcaatc ccaccaataa tacaatgcca ttgatagtgg
420ctgacctgaa gcaattcggg gaggagacgc caagctcgac gatcaccgga gcttgaaaga
480ccaacgagac aagatgacag gcccgtcgca ccacgccact aactgcccta acagaaatcg
540gcctgaatag tgcgacgagt gtcccggttc tgggcctcca cgataagata agtcatgggc
600ttatcgcgtc atcggcgccg atctcgcgat cagctgaaac caatcattca atcaatttgc
660atcacccgac tgggggcgag atttcagggc cagctgaaac ggtcggctgc cgagattgtc
720agtggatgat gaatgttatg ctggaagaga gggggagaat gacgtctcaa ttctgggtca
780cttactagtt gactagccac ctagtattta gctgctagct agggattcgg tttaaaagcc
840tggtggtttc tctcttcttc tcgtcatttt ctcttcatct catacccatt cttcaaaact
900cctccacttt gatcaattat cctccatcat ggctaccaaa atcaagctca tccccaatct
960caactacaag cgctcaggca ccaagtccta cgtgcacttg atgcgcaagt accgcttcca
1020tcccaccaag cctggtccct acactctcag cagctccatc caacagaccg gtcgtccgta
1080cactgaaaag cccatcgggg gtcgggccca tatccggcag ctggtgcgga agaagagcac
1140caccagcgat gaggttggcg aggttccggc cgaagatgtg cagaacgact ccatgtatct
1200ggcgaccgtg gggatcggaa ccccggcgca gaacctgaag ttggactttg acactggttc
1260agctgatctt tgggtacacc cccattatga aagacctaat atggaaacga gcgtcactga
1320cagatgtagg tctggtccaa caaactcccc tcaacccttc tatccgagaa caagacccat
1380gcgatcttcg actcgtccaa atcgagcacc ttcaagacct tggaaggtga atcctggcaa
1440atctcctacg gagatggatc ctccgcatca gggagtgtgg gcaccgacga cgtcaacatt
1500ggcggcgtag tcgtcaagaa ccaagccgtt gagctggcag agaagatgtc cagcacattc
1560gcccaaggcg aaggggacgg attgctcggt ctagcattca gcaacatcaa cacggtacag
1620ccaaagtccg tgaaaacgcc cgtcgagaac atgatcctgc aggatgacat tcccaagtcg
1680gctgagctgt tcacggccaa gctggatacc tggcgggaca ctgatgacga gtcgttttac
1740acctttggct tcattgacca ggatctggtg aagacggcag gtgaagaggt ctactacacc
1800cctgtcgata acagtcaagg cttctggcta ttcaactcga cctccgcgac ggtaaatgga
1860aagaccatta accggtcggg taacaccgcc attgctgata ccggtacgac gctggccttg
1920gtggacgatg acacgtgtga ggccatttat agtgcaattg acggcgccta ttatgatcag
1980gaagtacagg gctggatcta tccgaccgat acggcgcagg ataagctacc cactgtgtcg
2040tttgccgtgg gtgaaaagca gttcgtggtg cagaaggagg acctggcgtt ttcggaggcg
2100aagacgggct atgtctatgg aggaatccaa agtcgtggtg atatgaccat ggacatcttg
2160ggagacacat ttttgaagag tatttatgct gtaagtgcat tgctgttggc gttaaggggt
2220gatatcgaag ctcactaact ggattgcaga tctttgatgt cgggaacctg cgctttggag
2280ccgtccagcg cgaggagttg cgccagagcc tgaagtcgga gtagacgagt ggctgaatct
2340atcatggttt gatgatgtta tgtatttgca tgtcccgttt ttttggttaa atcctagtgg
2400cttttgagcc ccgaagtatg tacgcagtaa atagcagtgc aatatagtac tccgtacctt
2460gttatcagat gctctggttt gaagcacaaa ggaagccatc agtctcccaa tcagagcttc
2520cagcccgccc cgtccttcca tttccacaac ctcagcgcca tgaaacatgc catccctgct
2580cctaccataa ccgccgcaaa aagctgcccc ttccagaaaa cagcatgttc cagatccaac
2640aacgccccgg caataggatt tccaatgaga atgccaagcc ccgcgaacac gaaagacatg
2700cccatccacg tccccacaac agccatgtct ggacacaa
2738172349DNAAspergillus niger 17cgcgcactaa ccctccacgt attccaatat
accaaatctg cccaaagcgc cagccagctt 60cctcaagcct tgcggtcaga taaggccctg
tacctagcta gttgccgctg ctcccggcgc 120tgggccaagc cgtcggacgt ccgtcccccc
tctttccccc tcctctcccc tctccactgg 180tggaacgatg tctggctgtt gccatcgttc
tcagaagcaa cgccccctgg atcgggtggc 240tgtcgtacta ttgcatgttc gtccgcgcta
ctaggaaagt ttttttccca cccggagtat 300ccgtgtttag tccgcgggct ggctgaccgg
ctagctggcc gtgccagttg ggtaaggttc 360caagggagga ccttactagg tagaaacggg
atccaacaat gaggggaaaa gggcggatat 420ggcttgccgg gggttcattg cggcctggac
gaagaaaggg agatgatcac taatgcaaca 480caatcttggc ttgcaaggaa ttgcgctcca
accagaatgt ctctgcgtag ggatgccaat 540tcgtgcgggc catgctggat ggatagtacg
ctgctccact ctcgctcgac cttttgcagt 600ccacaatcgt ttccccgtat cgttgggcgg
gggcgttttt ctgcagctat ggttgctgct 660gccccgacgg tgaacctttc tgcatccccg
gttttagtcg attttagttg gcgggcctgg 720agattaaact ccgtcggacg aagaggagca
gtggtgtcat cgtcggcgga ttgcatgcta 780tcggaagagc atggaagagg gaaaacatca
acttcatttg caaaacgctc gagcataaat 840agaggcctgg attccgccgt tctggtgtct
tttcttcttc atccagcatc gcaagtctct 900caagcatcgc ctggttcgtt cttctcactc
ttccaccacc agccttgtca ataagttagc 960tcttcatctt ttcgaagaaa ccaattctcc
aaacgtcaaa atgaagttct ctaccatcct 1020taccggctcc ctcttcgcca ctgccgctct
ggctgctcct ctcactgaga agcgccgtgc 1080tcgcaaggag gcccgcgccg ctggcaagcg
ccacagcaac cctccctaca tccctggttc 1140cgacaaggag atcctcaagc tgaacggcac
ctccaacgag gattacagct ccaactgggc 1200tggtgccgtc ctgatcggcg acggctacac
caaggtcact ggcgagttca ctgtccccag 1260tgtctctgct ggatctagca gctccagtgg
ctacggcggt ggctacggct actacaagaa 1320caagagacaa tccgaggagt actgcgcctc
cgcttgggtt ggtatcgacg gtgacacctg 1380cgagaccgct attctccaga ctggtgtcga
cttctgctac gaggatggcc agacttccta 1440cgatgcctgg tacgagtggt accccgacta
cgcctacgac ttcaacgaca tcaccatctc 1500cgagggtgac accatcaagg tcactgtcga
ggccaccagc aagagcagcg gtagcgccac 1560cgttgagaac ctgaccactg gccagtccgt
cacccacacc ttcagcggca acgtcgaggg 1620tgacctttgc gagaccaacg ccgagtggat
cgtcgaggac ttcgagtctg gtgactctct 1680tgtggctttc gctgacttcg gctccgttac
cttcaccaat gctgaggcta ccagcgacgg 1740ttccactgtc ggcccctctg acgctaccgt
tatggacatt gagcaggatg gcaccgtcct 1800caccgagacc tccgtctctg gcgacagcgt
cactgtcacc tacgtttaaa tgcatctcta 1860tgcatgagat atcggtcgct tcaatgtctt
cgagacgaag acaaaccctg gggatgaatg 1920aaaaaatgag tgatgagcta tccggattga
tctgatcttg ttgagttgtt aattccgttt 1980ctgttgatgt ttttgaatga ttatacctac
ttttaagtag aagaaatgga tgagcgcgtg 2040catgctgaaa atgactgtcc ctgcttatat
tgtagaagat cttccagaaa gctgtgctgc 2100cgatctgaag atctgaagat cactagtgag
atctcgcagc tcggctgtgt aagtgcattt 2160gctctgtcga tcataacttt gtaaaagctt
gtatgcatag cagacatctg tcgattattt 2220agatgcttcg atttgatcat ctttactaga
atccccattc gagtagagct tcagagcgtc 2280gggtggaaat atcgggtcgt ggatggtatc
ggagaagtct cacaacatga acgaaagatc 2340cgcggtata
2349181495DNAAspergillus niger
18tgattgtgca gacttcctca tcatcgaagg agacaacaat gaaaccccag ggagacacgg
60aagcaggaaa acagactgcg gccggcgaga aagacacgat gtgacgaagg cttatctcgt
120gacgccaaga caatgcgggg aaagctgccc ttccaagacc caactggcct gcctctttct
180cattacaacc tcatcgcctt gattgacttt ctggaccttg gtggcttgga ggccggtcgc
240atcgagcttg ctactcgcta cttgctttgt ttcgatgact ggctttcgtt gatcgcaaca
300ttctttcttg gtatttttgt agggatagct agttgcttat catgggagat tacggccccg
360gagtgtcgtc actcacggca cagctacctg gaaatccgcc tgtctctgaa acagatcagg
420atgagatctc agtacttgta acgggctttg gggtaagctt ccagtgtcca taccattctc
480ccaatatcgc aactcacatg ttctacagcc attcaagtct aatctagtga acgcctcata
540tctgatagcc tcgtccctac caccctcttt cacattctca cctgcatctt cagacggctc
600tgatgctgtt ccccgtcgag tttcgataaa tgtccatcct tcacccatac ccgttgcata
660ttcatcggtg cggacgaccc tccccgtcat tctcgatgac tatgccaaga cgcacggagg
720ccgacgccca gacatcgtca tacacattgg catagcagca atgaggaact actattccgt
780ggagacgcag gctcaccgtg atgggtatct gatgtccgac atcaaaggca gatccgggta
840cgaggatggc gagaagctgt ggagggagct cgacttgcca ctggtgctta gggctggccc
900ttcagaggga cacgcctcgg agaagaaaca tctcagcccc cgtccaccgg acgaagattt
960cctagcagca tggaagacat tttgccctcc agaaaccgat gcgcggatct ccactgatgc
1020cggacgttat ctctgcgagt tcatcctgta caccagcttg gcactggcat accaggcggg
1080tgaggatcgc aatgtcacct tcttccatgt tcccgcgtca tgcttggatg aggatataga
1140gacgggcaag gaggttgccg tcgcgctaat caaggctctt gtgactagct ggagtgagca
1200gcagcacagc gttccctagt tctgaatgac tttttcaatc ttctcggagt tgtgacattg
1260catgtcccag cagttttggt ttatcggtgg tcctatctga gtactatatt tttttggcaa
1320tattttgcat tagtgaatac atatgggcgc ctgatgggtt atgatcgcat cacacctggc
1380gccgcataga ttttgggcat aaggagtgtt ggtgtaagat caccatcaat ttattactgg
1440tgttggattt gtgtagggat agaaccaata tagattaaat tctcacgcca tacat
1495192501DNAAspergillus niger 19gcgtcttcgg caagcggaag ggaccccgca
aaagctacgc attgccattg ttcaggaacc 60cactaaataa acacacggac atgtggtttt
ggctgcccgt gcgccccgaa gccgcagccc 120gaacggaggc attaaccctg atgcgtccgt
tgattgttat ggtcttgtcc aacataacgg 180ggaatttctg accgcacacg atgccaggca
acccatttta tcacggctga gcatgttgat 240aggtgttgaa ttcattttcc gtaggcagac
gtctcctaga gcaatttctg ctgtatggca 300gacattcgtg accgaaaccg tctcactcac
ggtctaccat ggtagttatg cttcgtacat 360aagacaagaa aggaatgcct tcaatcatgg
caagaaggga atttggtaat ttgggtgcta 420ctgccgtctt cctttgttgt gaaccttctc
cattacacac agtgtccctt gaaggccccg 480tatgccttca tactccgcca caatagccgc
catcggggct ttcctgcttt cgcagcccgt 540catgggctca aggcagggaa aggccccctt
tggctggggt actcagtcac ttgctcactt 600tggtatcaac ccagaccttg ggttgcacaa
ccagcagaac ctcaactccc tcatttcaca 660ttcagcgatg gccactgcgt tggagacgga
atatgccacc gtatgaaaag caccagccct 720ggctcttagt cctgatcact gattgtagac
tcagatccct attgaccata acaacgcatc 780ggctggcact tatcaaaatc ggttctgggt
cagcgatgaa ttctatcagc ctggcaaccc 840gatatttgtg tacgataccg gggagtcgga
tggcggatcg atagcccagt cctacctaac 900ctccactctc tccttcttca gagaattcct
gatcgaattc aacgccatgg gaatcgcctg 960ggagcacaga tactatggaa actcgacccc
ggctcccgta tcctatgaaa ctccacccga 1020ggcatggcaa tacctcacca ccaagcaggc
gctcgcggac cttccgtact ttgctagtaa 1080ctttagccgc gagaagtatc ctgacatgga
cctgacgccg cagggcacgc cgtggatcat 1140ggtgggcggc tcgtacgcag ggattcgtgc
tgcattaact cgcaaggagt acccagagac 1200gatattcgca gccttttcct catcgtctcc
ggtggaagca caggtcaata tgagcgcgta 1260ttacgaccaa gtctatcgtg gcatggttgc
cagcggatgg accaactgct cggcagatat 1320ccacgctgct ctggaatata ttgacgatca
actttcggat gaagatacag ctacctcggt 1380caaacaactt ttcttcggat ctggcgccga
gaccaactcc aacggtgatt tcactgcagc 1440gctaactgcc atctacggct acttccaaag
ttatggtatg gcgggaggta ttggaggtct 1500aggcgcattc tgcgagtatc tcgaaattga
tcccaagacg aacgggacta caggaccgga 1560tggccttgcc cctacgtatg gcggccagta
tgtcgccgaa cgatgggccg catggccaac 1620ctttctcgag ctggtcaatc tgaatatggg
gaccaactgc gggcctcagg acgcgtctca 1680gccaattgac tgtgactttt ccaagccata
cggcgatccc tcggccatca cttggacttg 1740gcaatactgc agcgaatggg ggttcttcca
ggcgaacaac gatgggccgc actcgctggc 1800ctcgcgatat cagtcggtgg aataccagca
agaagtatgt aaccggcagt tccccgatgc 1860agtggacaag ggactgctgc ctccgtcgcc
gcgggcggat gatgtcaacc aagagtttgg 1920gggatggacg atccgcccgt ccaatgttta
cttcagcgga ggagaattcg atccgtggcg 1980atcattgtcc attctgtcga cagaagattt
cgcacctcaa ggggtggagt ttacgagcgc 2040gatcccagcc tgtggggtgc agaccaatga
ggacaccgtc tttggatacg tcatgcagaa 2100ctcggaacat tgctttgact ttcaagcgac
gccgaccgtg gggaagttat cacgcggcat 2160cttcacatcc gccttgttgc aatggctcga
atgttttgga cagaactcaa gccaatccag 2220gtgatctggg gggccagggg gtgtgtgggt
gtgtaggtgt gggtgtgttt ggttgtgctg 2280gagcctggag ccaggataaa gaagatagga
caaggatgac tgagtggatc ctgggatgct 2340cctacttact tagaaaggta cagggcgcgt
cacggcgaag gtagactcgt accccagatt 2400agatgatcaa cgagaagcct cggaggtttt
ggatcaacgt tgggttttag cgtcacgtcc 2460aaagtcctgc atgcaggtgc cgcttttccc
ttttgggaga t 2501202660DNAAspergillus niger
20agagtccgcc ttgacaatgt ctaagaaagc atgctgagcc ccctccccca gtgtttatag
60gtgagtagga atggagtata gtgcttgagt gctttcacgc tggtaaacgg acccgctggg
120cggctagact gccttgtttt cggtctagga accattcaag attcaacggc tacaagtcaa
180gtcagaccat ctatcaatag tgcgagatag cagagagtcg gaccggagtc tcacaggcat
240ccttggcggt cgaggagtga tgagttgtgc gaccagcgcg tgttccttat ttaagccgcg
300cgcgcacctc tgacgcaacg gtctcaaagg ggggtctcac ctcttgcatg ttcggctgca
360ttcaattcga cctctccgtg tctgtctagc atcaccccaa atcctatcgg aatagtcctc
420cggttaagtc aacattgcaa atcagtcctt tcatttttgc ccagctcagc gctcggcctt
480ctgtgaccct tggagcctga tcaattgccg aaaggtgttc gggcgggtca gccttggaca
540gctcatttgc agtgcagagg ttcatcagca tcagctgtac tacttgactt cctatcatca
600ttattgatta ttatattata tcctatcttc tccgccaatc ttccatcgcc ggtagttttt
660cgtagacaac atgaagctct caatagctct tgcactcggc gcaacggctt cgacgggggt
720gttggctgct gttgtaccgc agcaagaacc gctgataacc ccccaagatc ccccaactca
780tcatcatcag gagaagttct tgatcgagtt ggctccttat cagacgagat gggttaccga
840ggaagaaaag tgggacttaa aactggtata gcaacattcc ccattttatt actgtcacaa
900tagcacatct cactgatcgc tctgcgaatg ttccattagg atggcgtgaa cttcatcgat
960attactgaag aacgaaacac tgggttctac ccaacgttgc atgctggtag ctatgttcac
1020tatccgccga cgatgaagca tgcggagaag gtggttcccc ttctgcgggg tctctccaag
1080gacaacatgg agcaaaacct caacaaattt acctcatttc acactcgcta ctataggtcg
1140tccactggta ttgagtccgc aaagtggcta tacagtaggg tttcggatgt cattgagcag
1200tcgggtgcag cagagtacgg cgccactgtg gagcagttcg ctcactcatg gggccaattc
1260agtatcattg ctcggatccc aggccagact aacaaaactg ttgtcctggg cgcacatcag
1320gacagcatca atcttttcct cccctccatc ctagctgcac ctggtgccga tgatgacgga
1380agtggaaccg tgactatact cgaagctttg cgtggtctgc tgcagtcaga cgccattgtc
1440cggggcaacg cttccaacac aatcgaattc cactggtact cggcagagga aggtggtatg
1500cttggttcgc aagccatatt ctctcaatat aagagagata agcgagacat caaggcgatg
1560cttcaacagg atatgactgg ttatacccag ggagctctgg acgccggtcg tcaagaagcc
1620attgggatta tggttgacta cgttgatgag ggactgacac aattcctcaa agatgtcact
1680actgaggtaa ggtcactccc gctttccttc tttgtgagac atataactaa cgattgcggt
1740caaagtattg tggtattggc tacatcgaaa ccagatgtgg ctacgcctgt tcggaccaca
1800cgtccgcaag caaatatggc tatcccgcag ctatggcgac ggaatccgaa atggaaaaca
1860gcaacaagag gatccacacg actgatgaca gcatccggta tctaagcttc gatcatatgc
1920tggagcatgc gaggttgaca cttggcttcg cttacgagct ggcctttgct caattctagt
1980gtccttcatg attttacgtt gtaaccgggt ctagcagata attctggcta actagtgagg
2040cttatatgtg ttcaggtttc ctatgtcggt ttattgtggc attagacaag attacagagt
2100aaccaatact ccattttgta atggaataga tgtctgaggc ccaagtgttg gcaaggaacc
2160ttgcctatat accacagatt aatactattg gctttggtga ttcaagaata ggcctatggc
2220tggactttat atatgttgac attctatttt gtaccggcaa cagactctaa cactagcacc
2280taacaaataa aacacgtgtg actcttatgg gtggcatgat gatagtctgc agaatggccc
2340gataagataa acgaaggaga tcctactata cccgattgag tcacactagc ctgaaacaga
2400aagacggtca ctccgccggg gcgatcactt ccactggctt caatcttcga gtcattcctg
2460cggtctgttt tctccttgtt accactccct ttgcccttca gatcagatcc tgaacggaga
2520aagctcaccc ttcatactac ataatattct acataacttg cttggactga attggcgata
2580ttccgtcggt cgctcctatc tatcgcctac gtcaccagcg gtgcccctcc aaggaagacc
2640ccaccatcag accttggatc
2660212047DNAAspergillus niger 21ccgagctata caaatgcgca tacaaaaatg
gacagagtta catttcaacg ataaacatga 60atgataagct tccagcaatg gcggccaagc
gatatgcaaa ccgattttag tacgcattcg 120gtttatctgt tatctctctc ggagtaaact
gtatgctctt cgccgaaacc acaaaagtaa 180gcggggaaac gtatactacc aatcaaacac
cccgagatgg tgactccgaa cgcagcagac 240ctatttagat atatttaggc atgacaagta
tcaatggatg acagaaacac cgagacctgg 300tacaatctaa gttgaggaga gatttgaatc
agaaagcagc tcgcacattt catcatgaga 360actactacgt cttttgctag gcttgcattg
gcagtggcct cagttggtat tgtctttgct 420agtccaacaa aaaataacga tgggaaactg
gtatatggct caccagaatc cgtcggcatg 480atatccgccc ctttgcacca aatggtccaa
aatgttagcg catatacaca tgctgccaac 540tatagcaagt tctcgtacga caaagtccat
cccatcgagc cagggtctgt taccctggtg 600gctctcgacg gtgtcatcgt cagcgaattt
gccttgggca agagaaatct ctacgccgat 660gtcaacggca ccaatttacc tcgatacctg
caggaagaca ccaccctgga tacagtctac 720gatatggcaa gcctcacgaa gctgttcacc
acggtagctg ctttacggga acttgacgct 780ggtcgaattg cgcttaatgt aactgttgca
acttatatac cggactttgc gacgaatggg 840aaggagaata ttactatctt ggagctgttc
acgcatacaa gcggtttcgc ttctgatcca 900tcgccaccac ttttctctgc ttattatacg
acgtatgatg aacgcattaa agcaattttg 960acgcaaaaaa ttatcaatac ccccggcagc
acatacctct acttagatct caactttatg 1020tcgctgggcc tcgttatcga gaccgtaacg
ggacgtgccc tggatgatct tatttatgac 1080ttcaccagac cgcttgaaat gacatctacc
ttcttcaacc gcgggaatat cgaaggctct 1140acaccccagt cacccaacta cgaccgcaca
gccgtacaag aatttcagat cgcagccctc 1200ggaccctcag aaccacagcg tccacaacca
gtgcgcggca cagttcacga cgagaacgca 1260tggtccctag acggcgtatc aggtcatgca
ggtctattct ccactgtgcg cgatacagcg 1320acattctgcc agatgatcct caacaacggc
acatatgcag gccaacggat cctttctcga 1380acagcggtag acatgatttt cacaaacttc
aatgccaggt ttccggggga tgctcgtagt 1440ttagggtttg agttggatca gtattctact
gcgggaccga tggcgagttt gcaaactgcg 1500agtcacactg gatttactgg gactacgttg
gtgatggata ggacgtataa cgccttttgg 1560ttgcatttta gtaaccgggt gcatccgtct
agggcatggt ctagcaatac tattgtgaga 1620gaggctattg ggtattgggt tgggaagagc
ttggggttgg atgttgcgtt tgctctgttg 1680taatggttga gcggaggacg taccagatgg
gatcctggat tcattccttc gatacacttg 1740tatgtacacg aggaatgtat tcgaatgcaa
atacctcata tattacagaa tccgcagtat 1800gaatccggat aattttgtta aggcaaatcc
agaaatttct aaaggttcac taccataaac 1860acaaatattc accattcaat atggcatgtt
atcatccatt ctcataaaag accctcgatc 1920cggaagaccg atcctgtcag ggaattagtc
aaccctctcc aataaatgca gcatgaaaga 1980aatctaccat gaatcaaccc cgagccctaa
ataatccgaa aagatccagt ctggtgacag 2040tgcaggg
2047222730DNAAspergillus niger
22tggccacgcc atcctcttat gctctctcta tccactctct ccctcagctt ctctcaacca
60cttccttacc catctttcac cctctcttcc cctttccccc tttcaaatct cttgaccttc
120aagtctggcg cgcgtctgga cagcggctct tgtgctagag gtcttaactc ctaccccgcc
180ttcctctggc cttgaccttc tctgttcctg aaccaggttc cctcccggtc ctctccgccg
240acgttatccc cgacattgtg ctgctcatta cctccgcccc caaggtaacg ggtaagcgaa
300aagtgcaagg cccagttggc taaatttcga tgactccttg agcgcgactc caagtccgct
360tgggaccgtt tcctccgctc cgtcttcggc aatatctact actactacta ttactattac
420taccccggtc agccgccgcc gccgtattac aagtacccat cgcctactga tggtctcctt
480atcattccgc ggggagacca cttccattcc ctccatggcg tcctggttgc tctcgacgct
540cctttttctg agcccgtcct tggtgtcagc caaatcggcc gcagactatt atgttcactc
600cttgcccggt gcccccgagg ggcccttgct gaagatgcat gccgggtaag cttcgctgtc
660ccggaacggt ccgcttagca cctatatact gacttattgc ctccacgcgc agccatattg
720aggtggatcc acagaacaat ggaaatcttt tcttctggca ctaccagaat cgccatattg
780ccaaccgcca gcggactgtg atctggttga acggtggtcc cggatgtagt tccatggacg
840gcgcgttgat ggaggtcggt ccgtatcgcc tgaaggacaa tgaaaccttg acctataatg
900agggttcctg ggacgaattc gccaatttgt tgttcgtcga tcagccagtc ggaaccgggt
960tcagttatgt caacacggac agctatcttc atgagctcga tgagatgtcg gctcagttca
1020ttgtctttct ggaagagtgg ttcagattat ttccggagta tgaacgcgat gatgtatgct
1080gcaatcacct atgctgccct agtcctgcac cttcacggat tctgttctaa catctgcgac
1140agatctacat tgccggcgag tcttacgccg gtcagcatat tccatacatc gccaaagcca
1200tccaggaacg gaacaagaac gttcaaggga agaccatcgc ttcgtggaat ctaaaaggcc
1260tattgattgg caatggttgg atttctccta atgaacagta catgtcctac ttgccctacg
1320catatgaaga aggccttatc aaggaaggca gccggaccgc gaaggaactc gaagttttac
1380agtcagtctg taagtccagg ctggaaactg gcaagaacaa ggtccacctc aacgactgcg
1440agaaggtcat gaatgctctg ttggataaga cggtcgaaga caacaaatgt ctcaacatgt
1500atgacatccg ccttcgtgac accaccgatg catgcggtat gaactggccc accgacctgg
1560aggacgtgaa gccctatctg cagcgggaag atgtggttaa agcgcttaac atcaatccgg
1620agaagaagtc tggctgggtg gagtgttcag gtgcagtgag cagcgctttc aatccgcaaa
1680agtccccgcc ctcggttcaa ctacttcccg gcttgctgga atcgggactt caaatcctcc
1740ttttcagcgg agacaaggac ctgatttgca accatgttgg aacggaacag ctcatcaata
1800acatgaagtg gaacggaggc acgggtttcg agacctcacc tggcgtctgg gctcctcgac
1860acgactggag tttcgaaggc gagccggcgg gtatctatca atatgccaga aacctgactt
1920acgtgctcat ctacaacgca agccatatgg ttccctacga ccttcctcgt cagagccggg
1980acatgctaga tcgcttcatg aatgtcgata tcgcgagcat cggaggcagc cccgccgact
2040cgcgcattga cggcgagaag ctgccccaga cgtcggtggg cggccatccc aacagcaccg
2100cggcggagga gcaggagaag gagaggatca aggagacgga atggaaagcc tacgccaagt
2160caggcgaagc cgttctcctc gtcgtcatta tcggtgtatt agtttggggc ttcttcatct
2220ggcgcagccg ccggcgtcac cagggatacc ggggcgtctg gcataaggac atgagcggaa
2280gctctgttct cgagcggttc cacaacaagc gcacgggagg cgcagacgtc gaagcggggg
2340atttcgacga ggcggagctc gatgaccttc attctccaga cctcgaaaga gaacactacg
2400ccgtgggcga ggacagcgac gaggatgata tttcacgaca gcattctcaa caggcctccc
2460gagccggggg cagtcataat ctatcctagt tcatctttgg ttgggtaaac ttgtgatggt
2520gtaggtgtat ggcttgcttg gggtctttgc cttttgtttt tgttttcttg gccacgaaag
2580gacgtgcctc cttatatgtg catttatcat tttatctagc tggctgcctg gcatcattta
2640acatttagac atgaacaaag tttatgaccg cttcctgata catagccagg cggaaattcc
2700tcccgtctat aagccaacag tctcatccac
2730232660DNAAspergillus niger 23actttattgc caagcgaatg acttcgactt
tcttatttct gcaacgcgtt tggccagtta 60ataatttgct agactcaact gattgctcgg
tgccatctta ccagggatcg gcaagccgaa 120aagtacgcta ttgcacccaa tctgtaaaga
agacttctcc aaactccaaa gtagaacaga 180cccatcttta gctttaataa ctccaatcat
gacgattgga actatcggaa aatataatct 240aagttcaatg gatctggtcc gtcgctagaa
tatcggagta aatgagccca acccctggcc 300atcaggttcg acggaggaat tcttctggtt
gggtacaaga ttgacacgat ccccttggcc 360gctgaacgtc actcttccgc tgtcaaagct
tgttggctga cttcgatccc ggatgatgtt 420tctgatttca cctgcagtga cagttgcggc
tgcacttctg ctgatcaacg gcgcaggagc 480aactcaatct gaacgaagtc gggctgccgc
tcatttttcc aaacgtcatc cgacgtaccg 540tgctgcgacc agagcccagt cgagcaacac
ttccgactac cgattcttca ataataggac 600caagcgtatg tattacaccg gccatgaact
ctattcgtcg ctgacatgtg gtcatccctg 660gatagcccac ttggtggaaa gcttacccga
tgtgcacttc gatgttgggg agatgtactc 720ggggtcgatc cctatcgatg acagcaacaa
tggatctcga tccctgtttt atatcttcca 780acctaagata ggcgaacctt cagacgacct
taccatttac ctcaatggag ggccaggctg 840ttcctccgaa cagggattct ttcaggaaaa
tggcaggttc acatggcagc ctggtaccta 900tgcacccgtc atcaacgaat attcttgggt
caatttgacg aacatgctat ggtacgcttc 960ttcctgagga ttaaattgga ggtatgcact
ctgtacattt tatgagacta atgtgaataa 1020attcagggtt gaccaaccag tcggaaccgg
attttccgtt ggaaatgtta cagccaccaa 1080cgaagaagag attgccgccg attttctcga
cttctttgaa aagtttgaag atctatacgg 1140gataaagaac tttcgcattt tcatgaccgg
tgagagctac gccggtcgct atgttcccta 1200tatctcgtcg gcaatgctag acaagaacga
caccacgcgt ttcaatctga gcggtacgag 1260ccctgtctac tcataactac tacccctata
agcttagttg actgaaactg actttttaac 1320gtccctctag gagcccttct ttatgacgcc
tgcatcggcc aatgggacta catccaggcc 1380gaactccctg cctacccctt cgtcaagcag
cacgcttcac tattcaactt caatcagtcc 1440tacatgaacg agcttgaaac cacctacgaa
gaatgcggct acaaggccta cttcgatgag 1500tactttgcct ttccaccaag cggcatccaa
cccccaaaat acatgaacta ctccgagtgc 1560gacatctata acatgatcta ctacgaagcc
tataacccga acccatgctt caatccctac 1620cgcgtcattg atgagtgtcc acttctctgg
gacgtcctgg gctggccgac agacttggca 1680tacgagcctg cgcccaccac atacttcaac
cgtatcgatg tcaagaaggc cctgcacgcc 1740cccatggatg tggaatggga gctctgcagc
tacgacctcg tcttcgctgg aggcgacgct 1800gacccgggtc cggagcagca aggggatgac
tcacccaacc ccaccgaggg tgtcctcccg 1860cgtgttattg aggcgaccaa ccgcgtgctc
attgccaacg gtgactggga ctacctgatt 1920atcaccaacg gcaccctcct cgccatccag
aatatgacct ggaacggcca gctgggcttc 1980cagtccgcac ctgccacacc gatcgatatt
cagatgcccg atctccagtg ggttgagatt 2040tttgaggccc aggagggata tggagggctg
gatggccctc agggggttat gggtgtacaa 2100cattatgagc gcggtttgat gtgggcggag
acatatcagt cggggcataa gcaggctcag 2160gatcagggcc gtgtctcgta tcgccatctg
cagtggctgt tggggcaagt tgagattctt 2220tagtctcccg ccttagatat aattgatatc
acactgtaat gttccctcca aaaagcttca 2280tttattgata gtttcagtga attttgttac
ttccagtaac aactctatta tggcattccc 2340tggtttttaa tgctgcctcc ttttaccgta
gctacggcta tgactacata taacggactg 2400atttttctat ctcaacttaa ctttgtattg
ccccctggat cctgactaca gaaaataaat 2460aattcaaatt gatatttatt ggccttttat
tggcagcata atatctttct catataagca 2520cgcgagaaac ttgttcctca tcatactata
tttatcccca cagctgatgc cgtgattcgg 2580taatgaagat ggagattatg cggattgcgc
acttcataat caaacatcgt ataagtgtgt 2640cagtaatata tatctataat
2660242800DNAAspergillus niger
24aatgccgaag cttgacctga tacgacttca aggtatcgtc accgacaatc gttatcatca
60cgctacaggc ccgcagtttc cgcttgaatt cccgcattag gaaatgagca tcgcattcct
120cttcccacga ggtctctttc cgagggcagc cgctgcaaca tcattgggat catgcttggt
180tctcctctcc catagctgtc cgcgagcttc tcattggtac ctcttcgcta cctcgttgca
240tcctattcgc gcatggcccc gccagagatg tttctgcaag gtcccatcac cttgccgcgt
300tgctattccc cgccctcgag ttcccgacaa gttactttgt gtcagtggct gagaagcctg
360gttctgagag tgtactcaga caatcatatg gttccctcca tgtgctacgt cgtcctagcg
420tcgctgcact acatcatcgt taggcagcat ggaactggca cccgcacata aagcccccga
480cacccccatc gataggctcg gtgttcgtgc acgcctgtcc actggcccct cccccaaagg
540cccttcatca gtatgctgtt tcgcagtctg ttgtcgacgg ctgtcctagc cgtctcgctg
600tgcacggata atgcttcagc tgctaaacat ggtcgatttg gccaaaaagc tcgcgacgcc
660atgaacatcg cgaagcgttc cgctaacgcc gtgaaacact cgttgaagat ccctgtcgag
720gactatcagt tcttgaacaa caagactaag cgtatgtatc tcagttcgat attgaacgat
780ggctgatttg cttccgtcgg acagcttacc gcgtggaaag cctgcctgat gttcacttcg
840atctgggcga gatgtattcc ggcttggtcc ctattgagaa gggcaacgtg tcacggtccc
900ttttctttgt cttccagccc actattggcg agcctgtgga tgagatcacc atctggctga
960atggtggccc tggttgcagt tcccttgagg cctttctcca ggagaatggt agattcgtgt
1020ggcagcctgg aacctaccag cctgttgaga acccatactc gtgggtgaat ctcaccaatg
1080ttctgtggta agtgtgatat tactggatcg ctagttgagt ttacatgggc ggtatcgacc
1140taacctattt tttgtagggt tgaccaacct gtgggaacgg gattctctct gggtgtccca
1200accgctacgt ccgaggagga gattgctgaa gactttgtga agttcttcaa gaactggcag
1260cagatctttg ggatcaaaaa cttcaagatc tatgttactg gagaaagtta tgcgggccgt
1320tatgttcctt acatatccgc tgctttccta gatcagaatg atacagaaca cttcaaccta
1380aaaggtgagt tatacttcac caaagtaatc tttaactagg gcttgtactg attgtactat
1440ctaggtgcac tggcatatga tccctgtatt ggtcagtttg actacgtgca ggaggaagca
1500cctgttgttc cctttgtcca gaagaacaat gccctcttca atttcaatgc aagctttttg
1560gcggaactag agagcatcca tgagcaatgt ggatacaagg atttcatcga ccagtatcta
1620gtcttcccag catccggtgt ccagccgcca aaggctatga actggagcga tcccacctgt
1680gatgtttatg acatcgttaa taacgccgtc ctggatccca acccgtgctt caacccctac
1740gaaatcaacg agatgtgccc cattctctgg gacgttcttg gattccccac cgaagtcgac
1800tatctccctg cgggcgccag catctacttt gaccgcgctg atgttaagcg tgccatgcac
1860gctcctaaca tcacctggtc cgagtgctcg gtggagagcg tctttgtcgg gggcgacggc
1920ggtcccgagc aggagggcga ctactcggcc aaccccatcg agcatgtctt gccccaggtc
1980atcgaaggca ccaaccgagt tctgatcggt aacggtgatt atgacatggt catccttacc
2040aacggcaccc ttctctcgat ccagaacatg acatggaatg gaaagcttgg attcgacacg
2100gcccccagca cccccatcaa catcgacatc cctgacctga tgtacaatga agtgttcatt
2160gagaacggct atgacccaca aggtggtcag ggtgtcatgg gcatccagca ctatgagcgt
2220ggtcttatgt gggctgagac cttccagagc ggacacatgc agccccaatt ccaacccaga
2280gtgtcatacc gtcaccttga gtggctgctt ggccggcggg ataccctgta aggcgggtag
2340gctaccacgg gggacgatgt cacgatgata gtcataagtt atgatctgta gatacgttgt
2400atgcgaatgt acatgaattg cttttactgg cagtctctaa agcaaaattc atagtagagt
2460actggcctac ttaccctcac ttcccctatc ttttcaacct gaagaccgga agaattgtaa
2520ctaacaagca taacgtagct gatttgaagc agagcataac acactctacc cctcggcact
2580tctacttatg acgctatttg actgctaact cgggtttaat cctgaagctg cagtccaatc
2640gtacattaaa ctcaatgtgc cttgcccagg aaacgatatt tgacttatat gatctgaaaa
2700tgaacaattg tccccgagag agagagagag agcgagcggt aaatacttag caagtcagtc
2760acgcagtatc ctccactaat gccgtaacac aggaaatgga
2800252165DNAAspergillus niger 25gaatcaggat ggaggcccga cggccatcaa
ccgccgaaca tccggctgct cgaccgaatt 60cggagacatg gtcggagcta ggcgggaata
gccgtccaaa gtcacgatag gtcgcttgta 120ctgtgtagaa gtcagaccct ttggatccag
cgttcgttgt gaatggctcc acgcagcctt 180tgctttgacc aatttggggc tgagtcactt
ggtcgatgat gactggctat accagtgcaa 240agtgtgagca ccatgcttac actattgcct
tagcctggcc gttgaagctg acatgagagg 300aacaggcaga agagaaatgc tgccttgcct
cttgggggcc aatttaacct ccccagactg 360cggcacatcc gtcttggact tgtgcttctt
tgcaacttac gttataacga gtctgtgaac 420aggaaaacag acgcccggtc agtcaagatg
aaaggtgcgg cgctaattcc tcttgcggcg 480ggcattcctt ttgcccatgg cctgtctctc
cataaacgcg acgggcctgc cgtcgttcgt 540atgcccattg agcgcaggag cgcccagtcc
ttgcagaaac gagattctac ggtcggtgtg 600actttgcaga actgggtatg ttaagctata
cggccgtgaa gtatagatca tgctgacaat 660cgctaggatg cgacctatta cgcagtcaac
ctgacgttag gaacacctgc gcaaaaggta 720tcattagctt tggacactgg cagcagcgac
ctctgggtga acaccggcaa ctcaacttac 780tgctcaatcg acaatctatg caccccttat
ggcttgtaca atgccagcga atcgtctact 840gtaaagaccg tgggcacaca cctcaacgat
acatatgcgg acggcacaaa cctttacggt 900ccttatgtga ccgataagct cacgatcggc
aacacaacaa tcgataatat gcagtttggg 960atcgccgagt caacgactag taaacgtggg
tgaaccgttt gtccatgaat gatcgtcgct 1020gactaggtct actataggcg ggatcgccgg
cgtcggttac aagatttcga cctaccaagc 1080cgagcatgac gacaaagtct acgccaacct
ccctcaggcc ctcgtcgaca gcggtgccat 1140taagtctgct gcgtacagca tatggctaga
tagtttggag gcgtcgactg gctccctcct 1200tttcggaggt gtcaatacag ccaagtacaa
gggcgatctg cagactcttc cgatcattcc 1260tgtgtatggc aaatactact ccctcgccat
cgcccttacg gagctcagcg ttgcgaccga 1320ctccaactcc agtagcttca ccgacagtct
ccccctctct gtgtcactcg atactggcac 1380caccatgacg gcactgccca gcgacctggt
caacaaggtc tacgatgcgc tcaacgcaac 1440ctacgacaag acatacgaca tggcctacat
cgactgcgac actagagagg cggattacaa 1500tgtaacatac agtttctccg gggcaacgat
caccgtgagc atgagtgagc tgattatccc 1560cgcaacggag ccggggtggc ccgacaacac
gtgtgtcttg ggcctcgtgc ctagccagcc 1620gggcgtgaac ctgctcggtg atacattcct
gcgcagtgcg tacgtcgtgt atgatctcga 1680gaacaacgaa atctctctcg ccaataccaa
tttcaatcca ggcgacgatg atatcctcga 1740aatcggaacg ggaacgtctg ctgtgccagg
agccacaccg gttccctctg ctgtctcttc 1800tgcaactgga aatggactga tctcgtctgg
caccgcagtg cccacgctgt cgggtgtcac 1860aataactgct acagccacag caaccggctc
aaccggcact ggctctagcg gtggttcgtc 1920ggctgaagcc acgagtactt cctcggaggg
cgctgcggcg caagctacga gcaacccgat 1980gaacctgctc ccaggacttg cgggtatcgg
cctacttctc gctctgtaac gcgattgtac 2040ctactcaaat agatatcacg acgagactct
aatgtaataa tgtggtatac aataacccca 2100atatctacat ttccttaacc gtaaactgca
tactctacac caaatccacc accaaaatac 2160atacc
2165262800DNAAspergillus niger
26gcccagccaa tcataagaaa cccggtcctg gcaaggtctt ggcggggatg gtgcaagcca
60gccaaacaac cggtttgttg gacgccctgc ggtacactga aatcttgggc tgttcgacaa
120aaacaaagca ggtaacacaa agatataata cggcggaatg ataggatatc cctgtcgatc
180agccggacag gatggcgcta gtgtcacctc cagcttcggc taccctcgca gcggacccaa
240tcagcgtccc tcccagcccc cgatacagta atggtatgca catcgcagtc ttaatcgcct
300gcagcggcag taatgatagt cctgccggtg aataataccc cataacaaac aaataaataa
360tactacttat ctctcctcgt ccctttcact ttccctttgc cgtcttcaat cccctcatct
420tggtctcttc ggcagccttt caccatgctg tcgtctctcc ttagccaggg agcagccgta
480tccctcgcgg tgttgtcgct gctcccttcg cctgtagccg cggagatctt cgaaaagcta
540tccggcgtcc ccaatggtga gttatagacc ccaattcttc attttgagcc acatactgac
600gtgattcctt cgaatactac caggctggag atacgccaac aatcctcaag gcaacgaggt
660cattcgcttg caaatcgccc ttcagcagca tgatgtcgct ggtttcgaac aagccgtgat
720ggatatgtcc acccccggac acgccgacta tggaaagcat ttccgcaccc acgatgagat
780gaagcgcatg ttgctcccca gcgagactgc cgtcgactca gtccgcgact ggctggaatc
840cgccggtgtc cacaatatcc aggtcgacgc cgactgggtc aagttccata ccaccgtaaa
900caaggccaat gccctgctgg atgccgactt caagtggtat gtcagcgacg ccaagcatat
960tcgtcgtctg cgcaccctgc aatactccat ccccgacgcc ctggtctcgc acatcaacat
1020gatccagccc accacccgct ttggccagat ccagcccaac cgtgccacca tgcgcagcaa
1080gcccaagcac gccgatgaga cattcctcac cgcagccacc ctggcccaga acacctccca
1140ctgcgactcc atcatcacac cgcactgtct gaagcagctg tacaacatcg gtgactacca
1200ggccgatccc aagtccggca gcaagatcgg ctttgccagc taccttgagg aatacgcccg
1260gtatgccgat ctcgagaggt tcgagcagca cctggctccc aatgccatcg gccagaactt
1320cagcgtcgtc caattcaacg gcggcctcaa cgatcagctt tcatcgagtg acagcggcga
1380agccaacctc gacctgcagt acatcctggg cgtcagcgct cccgtcccca tcaccgagta
1440cagcaccggc ggacgcggcg aactagtccc cgacctgagc tcccccgacc ccaacgacaa
1500cagcaacgag ccctaccttg acttccttca gggaatcctc aagcttaaca actccgacct
1560cccacaagtc atctctacct cctacggtga agacgaacag gtatgcacct cacctgaccc
1620attccatttt acatccctca cctctctcaa ccaaactaac aacaccaaca gactatcccc
1680gtcccctacg cccgcaccgt ctgcaacctc tacgcccaac tcggcagccg cggcgtctct
1740gtaatcttct ccagcggcga ctccggcgtc ggcgccgcct gcctcaccaa cgacggcacc
1800aaccgcacgc acttccctcc tcaattcccc gcctcctgcc cctgggtaac ctccgtcggc
1860gcaacctcca agacctcccc cgagcaagcc gtctccttct cctccggcgg cttctccgac
1920ctctggcccc gcccctccta ccaacacgcc gccgtgcaaa cctacctcac caagcacctg
1980ggcaacaagt tctcggggct tttcaacgcc tccggccgcg ccttccccga cgtctccgcg
2040cagggcgtca actacgctgt ttacgacaag ggcatgcttg gccagttcga cgggacgagt
2100tgctccgcgc cgacgttcag tggcgtcatc gcgttgttga acgatgcgag actgagggcc
2160gggttgcctg tgatggggtt cttgaatccg ttcctgtatg gtgtcggaag tgagaagggt
2220gcgttgaatg atattgtgaa cggcgggagt gtgggttgtg atgggaggaa tcggttcggg
2280ggcacgccta atggtagtcc tgttgtgccg tttgctagtt ggaatgccac gaccgggtgg
2340gatcctgtgt cggggttggg aacgccggat tttgcgaagt tgaaaggggt ggcgttgggt
2400gaggagggtg gtaattaagt gtgagatggg gggaaaggga ttttcttttc gatgtgaata
2460ttaggtgaat tgtgtggata attttcatac ataattaagt ctgcattggc agtgataacc
2520tggaagaaat gtctaatgag tgtgatttgt ttacttatgt atattgagta atggaatgta
2580gatgacttgt ctttgtactg tataacgaaa tgattatttg agtggagggt attaaagaac
2640tataaaatat atacaaaggt taacccatgc agtcgtaacc cataatgcaa agctctactc
2700tatctgtatc ggtagcagat aagtgtatgc aatctatctt tgttgatgat gcaatcaagc
2760gggcacacca ccagtgccaa acagctccat cttgatgcgg
2800272660DNAAspergillus niger 27ggagacagaa tacatggaat tgatgctcaa
caacgacaag gagctcgcgt tccgtaatgc 60caagttcatc gcttagattg ggaggatcgg
ccaggtctgt atataggccg cgatagaaca 120cgaaataaat gatttatgac tatttgcggg
catgagccct cacgtagtgt gctatagacc 180gtagacttgg taaacattgc tatcgatccc
ccccttcgta tattagtcat aattcttcct 240gatgtcaact ctcaaaacac agcattccac
agcgttcata tgatgtcaat gtctcaatcc 300ctaacgtttt gctgaaacag gtacctcttg
gttatttttc atatagtagg aaggtcgccc 360ttgcatgtgg tatgagacca ttggcggcat
ctagcttgaa tggtactcaa tatgaccagt 420agtctccact tcagcagtgt gtttgtgata
gccaagacat cggcagtagt tcctgcagag 480cttctacacc tcttcatctt cacttcacca
atccactctc tcactgctca cttctagttt 540cgcaacacta gagaatcatg tggctctttc
tcgtgtgcag tatcctgctg ccacttggag 600tagtcaacgc acagtctcaa tacttcaaca
acaaaaccaa aggtatacca catcgcgtca 660gtttgaatcc cactaacagc gctagaattc
gtcgtcaatg gctctgctat tccttttgtc 720gatttcgaca ttggcgagtc ctatgcgggc
tacctaccca acacgccttc tggaatctcg 780agtctatact tctggttctt tccatcttct
gatcctgatg cgtctgatga ggtatgctta 840tcgccgtctc atatgcttcg cacggctaat
aaacagatca ccgtctggct gaatggcggc 900ccaggatgca gctctctggc aggcatcatg
ctcgagaacg gcccctttct atggcaacct 960ggtacctacc gacccgtgcg caacccttat
gcctggaaca acctcacaaa tatggtgtac 1020attgatcagc ctgctggaac gggattctcg
cttggcccgt ctacggtggt ctcagaattt 1080gatgtagcca gacagtttat ggacttctgg
aggcggttca tgaaaacatt cgatctgcag 1140aatcgaaaga tatatctcac tggcgagagc
tatgcgggcc agtacatccc atacatcgcg 1200tcgcagatgc ttgaccagga tgatgatgag
tatttccggg ttgccggcat ccagatcaat 1260gatccctaca tcaatgagct gccagttttg
caagatggta tgcctctgac acaggttatt 1320gcttactcct ctgacattga tgacttccag
tgcctgcagt tgcgaccgtc aatcagcacc 1380gctccctctt tccctttaat gacaccttca
tgagtcaaat caccaagctt tccgacgatt 1440gtggctacac ttcgtttctt gacgatgccc
ttacctttcc accccgttct caattcccat 1500cagtgcccta taatgctagc tgcaacatct
gggatatcat aaacaacgct tctctagctc 1560tcaacccatg cttcaaccgc taccatatcc
ccgacgcctg ccccaccccc tggaacccag 1620tcggcggccc catcgttgga cttggtccga
ccaactactt caaccgcagt gacgtccaga 1680aagccatcaa cgcgtaccca acggactatt
tcgtctgcaa ggatggaatc ttcccgacgg 1740ccaacggact ggacacatcc cctccaagct
ccctgggacc gctgccgcgc gtcatcgaac 1800agaccaacaa taccatcatt gcgcacggcc
tgatggattt cgagctgctg gcgcagggaa 1860ccctgatcag tatccagaat atgacctgga
atgggaagca ggggttcgag cgggagccgg 1920tggagccgtt gttcgtgccg tatggtggat
catcgggagg aggcgtgctg ggaacggcac 1980atacagagcg tggattgaca ttttcgacag
tatttagttc aggacatggt aggtccatat 2040ctcagtatgc gatgcagcgg gttgctaaca
ggaatagaaa tcccggaata tgcaccgggg 2100gcggcatatc gccagctgga gtttttgctg
gggagggttg cgaatctgtc ggtgggttga 2160ccggagacag caaagagaat gaagaaaaga
aaaaaaaaaa aaaaaaacaa tatgaataat 2220tgcagacaat tattgagcag gtgcagataa
cagagcagaa tggttgagct gtcagatcgg 2280ttgacttttc ggcaatgaga tttggggaag
ctagctctga gattggtcct ccgccaaagc 2340tatcgcaatc atgatccaat caacactgca
tactaccttg atcccctcgc tgtagacccg 2400tattgccgga caaacaattg actagatatg
aactcttgat tataaaccaa tcaagacccc 2460cagtagcgca tattatcacc cccatacgtt
catgatcggc agcctaatcc gaggtcggat 2520cgggaattcc ttcaaactct gaaaccgcgc
tgggtccacc ccgcatcttc acagtaaaaa 2580tctaagattc ccctcaatcc ctgctccatc
tactttcctc cctctcatac cacttttgct 2640ttttgtatag taaatatcca
2660281540DNAAspergillus niger
28taaacagaac taagcaaaag tccagatctt gtgtcccctt cgactaccaa tcggaacaat
60aggaattaac tattagcctg caactgtctg caccatcatg caacccctct cgttgtgatt
120gctcctgttt tacattcccc gagacatgct catctagttg caagccatct ctaacccacc
180tgcatgaagg gtgcagaaac acgaacagtg gcgtggtagc acacactctc tgcagtactc
240gatgccaata tttccaccag ctgaactaca atctccgcgt cagaattaat atataaccat
300caccaatacc ccttacatcc caactcagca actacagcaa tacttacacc tatcctccac
360aaccccacaa ctccacccca tcatgtccaa actctccgct gctatctcca aggtacttcc
420ccatatccac aacacccccc acaccatccc tcaaatctaa caacaataac aataacctct
480caccacccca gctctccctc tccaccatag ccaccactct gctcctcctt acccccccaa
540ccaccgccta cttctacaaa tatcccgccc tcttcgtcta caaagacacc aactgcaccg
600atatctcctt ctcacttgtc tacccctccc tgggtaactg caacggcgga tactacgact
660acgcgggctc attccagatg ttcaatatcg atgctgcgta tacctgtaat ggcagtgact
720cgacactgat gtttgagatg tataatagct ccggctcgga ttgtggagat gagagtgatt
780tgttgtttag acagccggtg acggaggagt gtactgttgc ggatgtggag agtccggggc
840cgttggagat gccggtttgg tttgagttgg ggtgtatgta attgattcgt tgagtcggta
900aggagggatg gacggggggt tatataggta gcttggtggg taagcactat tggggaattg
960tggtgggatg gctggtacta tgttgttcgg tgtggggatt cttgagggtg ggttagagac
1020taaagtgagt tgtctgaggc agtagtggat gacttctgct tctgtgatgc tggtatgtga
1080taaagttaat tacatatgca aatgtgcaat catagactga taggtggaaa cttgtctact
1140cgttgagata tatagttata ctggaaatgt tattcatcaa ggctgaatac aagtgtaacc
1200gtgcacagat tatctttgat actgtctatg ggctgtacga gcgtctctga ctcctacaat
1260gagttagcgg ctgcacatta ctatgaggac ctgtgaattc gacaccaaaa ataaccagcc
1320ctgcagccga ttacatctgc ctttccatct attttctgga gtcaagcagg tattccaaca
1380tactcttact tgcttaatac accaactttc atatatctac ccagaattac tcaatcgtga
1440tgtaggcact ggcaatcata gtaactatat ttcgtttcat tttatttgtt tgtacaattt
1500tttttttccg gtagtttaat cattccaatc aaagaattgt
1540292800DNAAspergillus niger 29acactttctt cgatgctatg tcccagccat
ataaggttca attctccgac tcaagtgaca 60gaatcagcta tacacggttc actgcttctt
atatagtaca tgaggattca ataaagccaa 120gttcagcaga gtcttggttc cagagcttga
atactcggta gcattccagg gattgtgggg 180agacaatcca ggagcagcag ccatcatttg
gtgcctgagg cacacagcac tgcctcgtct 240ccactcccaa tacttaccaa gtcacccacc
attccttccc tctttcggcc tttctctttc 300tctctctcac ttctgctctc gtgcaccact
tttcttccca ctcacgccat ccttctccat 360ccatttcctg ccaaactttt gtctgtcatt
ttaggttttt tgaaggagat gactcggtga 420ctactcttca atcccggcgt actcaacatc
cataatgcgt cacctcttat cactgctggt 480gcttctgatc gcatcggccg ccctggtctc
cgccgtcccc gccggctcca ttatcactcc 540acaaccaccc gtcgagcccg ttcaccttct
ctcttcccag ccctctgatc cccgaaggcc 600atggatccgc ctccgtgact ggatcatcga
gtccatatgg ggcatcgaaa aacccgcatc 660tcgtcgattc ccactcaacg attccccgcg
caatcgctct cctccctccc ggattctggc 720gcgctacggt agtgacgtcg tacttcgttt
cagcctgcgc aatcacgatg aggccgaggc 780attggcccag gctgcagaca ttctattcct
ggacgtatgg gcgtctactc cagcattcgt 840agatatccga ctggccgagg aagtcgtaag
tggttcatcc ttccgtccat atctgcccgg 900ttgactgatg cctgatccac tgtgaccctc
gcaacagatt ccctcattat tgggcttgct 960accaaattcc ctccagaccg catatactcc
cctaatagac aacctggcag agagaatcta 1020tacgacctat ccatctaaaa agccgatagg
acttgaagga caatctggat ttgcgtcctc 1080gagtcgacct gcgccaaagt tcggtgacct
ttttttccac gagtatcagc ctttgtccgt 1140cattatcccc tggatgcggc tgctggcttc
catgtttcca tcccatgtgc gcatgattag 1200cgttggagta tcttacgagg gtcgcgaaat
tcccgccctc cgactgagcg caggcagctc 1260caccgcggcg tcaggccctc gtaaaacaat
catcgttacg ggtggtagcc atgcccgcga 1320atggattggc acctcaaccg tgaaccatgt
aatgtacacg ctcattacca agtatggcaa 1380atccaaggcc gttacccgcc ttctacagga
cttcgactgg atcatgatcc ccacgatcaa 1440tcccgacggc tatgtttata cctgggagac
ggaccgacta tggcgcaaga atcgacagcg 1500gaccagccta cgcttctgtc ccggaatcga
tcttgaccgc gcctggggct tcgaatggga 1560cggcggtcgg acccgcgcta acccttgttc
agaaaactat gctggagacg agcccttcga 1620gggaatggaa gcacaacaat tagcacagtg
ggcgctcaac gagacacaaa acaacaatgc 1680cgacatcgtg agcttccttg accttcactc
ttactctcaa acaattctct accccttctc 1740ctactcctgc tcctcgatcc ctccaacgct
cgagagcctg gaagagctag gccttggcct 1800agccaaggcc attcggtacg cgactcacga
aatctacgat gtcacttctg cctgcgaagg 1860catcgtcacg gccagtgcgg cagataacaa
ccccgggcgg ttcttcccca ttggtggcaa 1920ctccggtggc agtgcgttgg actggtttta
ccaccaagtg cacgcgactt attcatacca 1980gatcaagctt cgtgatcgcg gaagctacgg
gttcctcctt ccgtctgaac acatcatccc 2040caccggcaag gagatctaca atgttgttct
gaaattggga tccttcctca tcggaggcga 2100ctcatttgac gtcgattggg aatcagaact
cttcgatctg tcaaaggacg aatccgatct 2160ggatagccgc tattcaaaat ccaatgaccg
ctccccggcg tatctacaca acgccaacgg 2220ccccctgccc aacattgacg aagacgaaga
taaggaatgg gtaatggtgg aggaagaaga 2280ctacacagac gatgacgacg acgatgatga
tgatgatgaa gaagaggaag aggaagagga 2340agatacatat tgggccaccg aacacacata
cgaatttcgg cgacgacgct gatgatggac 2400aaactaatca accctattta tatgacaccc
tcgcccatat actctccttc tgatgacgat 2460caataatgac taatgacggg tttgcgggag
tgattgacgt gtttatcatg ctctgcttct 2520cgattctttt tattatttta gatactgttc
ccttcaggtg cctatagcct atagagctta 2580ggggtattct ctacataatt aaatacagag
taattcaatg aatcgtccta tcaaataatg 2640gacccgagta actaactggt gtttcattcc
ttccgtactg tacttaatct gtaaggaatt 2700gaaagctaag cctcataagc tacttcaccc
cactctttac ccaggtgcag tgtatcatga 2760tcatcgtcat acattcattc gttaggtatc
tttccgatgc 2800302380DNAAspergillus niger
30cccgcctgta caaggctcat tgagcgacct ttatttctat gaaggcttct tgcagtgtag
60agccgctgtt tagaactcgg aaataggcgt gcatagtatg gacccaatca acagagttaa
120tctatttact ctaacgccta gcaagcaatc agtgcccaga ggaagctaac ggatggctgg
180ccaagctgcg ccagaaacga aatgagtccg taataccatc cctgcatgct tatctgtatt
240ctgtgcatgc atgatgcttt cctcatgggg cattacccag tagtccgaag acgcaatgtg
300accatctgac tgagttttaa atatactgtc caagtgcctt ctgacccggt ccccgcttga
360tggcaatcaa caaaaggtga atgtgactga aaggcgcggt ccagacaaca ggacttagac
420tttgttgtga gactataaaa ggatctaact attgcactac tgaaatcaag tattctagtc
480taccattgac atttctcccc tttcggtggc tactcgctca acatggcttt cctcaaacgc
540attctcccgc tgctggccct catcttacct gcagttttca gtgccacaga acaggtccct
600catccgacca tccagaccat cccggggaag tacattgtta ctttcaagtc cggcattgac
660aatgcgaaga ttgagtctca tgccgcatgg gtaacggagc tccacaggcg cagcttagaa
720ggccgcagta caaccgaaga tgaccttccc gccgggatcg aaagaacgta cagaattgcc
780aattttgctg ggtacgcggg gtctttcgat gagaaaacta tcgaggagat ccgcaaacat
840gaccatgttt gtgtccacgt atcctagacc gtatggtttc gactaattgc tgtacaggta
900gcttatgtgg aacaagatca ggtctggtat ctcgatacgc tagttaccga aaggcgagct
960ccttggggac tggggagcat ttctcaccgt ggtgggtcta gcaccgacta catctatgat
1020gacagcgctg gggagggtac atacgcttat gtagtggaca ccggcatctt ggctacgcat
1080aatgagtttg gtggtcgtgc tagcctggca tataatgctg cagggggtga gcacgttgat
1140gatgttggac atggtacaca tgtagcaggg accatcgggg gcaaaacata cggggtttcg
1200aaaaacgctc acctactgtc cgtgaaggtg tttgtaggtg aatccagctc gacatcggtc
1260attctggatg gcttcaattg ggccgccaat gatattgtga gcaagaaccg gaccagtaag
1320gcggcgataa atatgagtct tggtatgtgc gccctctctg gggatctaat cccgctaacc
1380gtgatgcagg tggaggctac tcctatgcgt ttaacaatgc agttgagaat gcttttgacg
1440agggtgtgct ctcttgtgtt gccgctggaa atgagaatgt aagctctgct gaactgtcca
1500ccattgagct aaatttagac taatgttttg cagagagatg cagcacggac tagcccggct
1560tctgcacccg acgccattac tgttgccgct atcaacagaa gcaatgcccg tgcgtcattc
1620tcaaactacg gctctgtggt tgacattttt gccccgggag agcaagtact ttctgcatgg
1680accggctcga actcggccac caacacgatc tccggcacgt ccatggctac accccatgtg
1740acaggtttga tcctctattt gatgggcttg cgggaccttg ctaccccagc ggctgcaacg
1800accgagctca agaggttggc tacgcggaat gctgtcacca atgtggcggg tagccccaat
1860cttctggcct acaatggaaa cagcggcgtg tcaaaagggg gtagcgatga tggagatgag
1920gactaggtgc gtaacatgag tgaatatggc ttagaatagt ggggatcgga gagtagacta
1980gtttatatgc gaaataaagt gtgtatcagc accctggcct gttcatgtaa gtcggcattt
2040tcacttttgc cgacaccgca aatatgctgt gcttgaggct gttgcctccc cagccagcct
2100tcccgagact gaaactcaca catccattgg atgtataaag ttctgcacat gcgaaatgcc
2160gctgccgttt acctcccgac gtggtaccgg accgaaggca gacacagatc atggaccgct
2220ataccgcaca gacaacttgt gctccttact gaaagtacca ttccacaggt cattgcagca
2280tgatgagtga tgatgtactt ctccccatca agaaccactg gcggtggttg gaatgaatct
2340agatcaaaga gatcaaccgc ttccccggac agatcaggcc
2380312441DNAAspergillus niger 31aaacgacgtt ttaggtcaat actgaagtcg
ttgaaaacgc ttgattcttc gctatctagg 60cgcgctcgga ggagcagttg aagttacgga
gttcgggtca cgtgacctcg actactagta 120atctactgag attacgttcc aacataattt
catcaggaag aatgcaaagg ccccagagaa 180ggagaatttc acgatcggtg aaaacgacca
agcagcatca catttgaatg aaactacatt 240gctccgtgtt tattccgttt ctctctcttc
tctctatcta ttgcttctct ttggctagac 300ttcaccaact aattaatgct tacacgatga
tagctgcatc cattgaatca aactgacccc 360gcaaagctga ctgaaccaac ccctccggtg
ctgttgctac tgccagtttg aattcaatat 420cctataaccc accctgctca atgatcaccc
ttttgtcggc cctgttcggc agcgtagtat 480atgccgctac gcagaccgtg ttagggccag
agggggctga tccctttacg gtgtttcgca 540gcccacactc accggcattt tcaattcgca
tccaggagca gaatgactcg atctgtgatg 600ctcgttcacc ccaattcact ggttggctcg
acattggccc gaagcatctt ttcttttggt 660attttgaaag ccagaatgac cccttccatg
atcccctaac gctatggatg actgggggcc 720caggagactc gagtatgatt ggacttttcg
aagaagttgg cccttgccgg attaatgagt 780ttgggaatgg aacagatcac aacccctggg
cctggaccaa gaattcatca cttctttttg 840ttgaccagcc agtcgatgtc gggttttcct
atatcgatga gggctatgag ctgcctcatg 900actcacgtga agccgcggtg gacatgcatc
ggttcttgcg attattcata tccgagattt 960ttcctcacaa acagttcctt cccgttcacc
tttccggtga atcttacgca gtaagataga 1020agccgccccc agaaaaagct tgctcagtgt
tctaacaatt accttgtgat acactgtgta 1080gggccggtac attccttatc tggcgaccca
aatcttggaa caaaatgaac tgtataaaga 1140tagccccagg ataccgctga aatcgtgctt
ggtgggtaac ggattcatgt cacccaagga 1200tgcaacgttc gggtattggg aaacactgtg
tactactaac tcaggagtcc catctcctat 1260cttcaatgaa actaggtgcg atattatggc
ggcgaatatg ccgcactgta tggatctata 1320tgacatatgc attcaacact cagaccccgc
gatatgtcat gcggcccagt ccgtctgtta 1380cgatagtgtt gtagggtagt atgataacga
ggccggcgct gatggccgta acagatttga 1440tagtgagttg ttatcatcag aggctcatgg
ccaaattatt gctaaggatg acgacagtca 1500ctgcaccttg tgagatcgac gaaatgtgct
atatcgaagc ggctctaatt gagagatatt 1560tgaattcgcc atctgtttgg gaggccctgt
cgccaccgca acaggttacc gaatacaaat 1620tcgtcgctac ttctgttatt gatgcatttg
ctcaatcagc ggacggcatg gtgtcgagct 1680cgaagcagat cgctttctta ctcgcaaata
atgttgactt cttagcgtat caaggcaacc 1740ttgatctcgc ctgtaatacg gctggcaacc
tacgttgggc gaactcgctt tcttggaaag 1800gccagacaga atttaccgca aagcccttac
ttccgtgggt ctcgatcaac tctgggagcc 1860aggaacctgt ggggagtgcg aaggaaattc
aggtttcggt cggtgaaggg acggacgaaa 1920cgtcacgctt tgcctttgtg actgtggaca
acgctggaca cctggtaagt caaagtaaaa 1980ttaggagaga ggggtttaat gatccagttg
gctaacgagt agtgttggta ctatggtagt 2040tgccccaaga tcggccggat gtagcgcttg
acatgatgat tcgctggatt actggggcat 2100cctttgttta acaaggaaca gttgcgggac
tcaaagattt caaactgacc tactccttac 2160cttttatgaa atgggcccgc tgcctaggtc
aaacaagggt gctcgggcaa acaataccag 2220ttttggacta ggggtccgtc tgctttcatc
agcccttcgt agcttggact tttgtccatg 2280tccctgctgc ctcggtccat cacacgcccg
ctccccccat ttcctttcct tcccttctct 2340tttcgttcta ttatgcttga gtgcccgatc
ttccctgcat gctatgccta tttcagtgta 2400cgcagtttgg gtttttgtcc gaggaagaca
atcatccatg c 2441323500DNAAspergillus niger
32gtttgttgtg gttgttgttt ggggtcccat ttgcaatccc tgaaccggac cctgtcaatc
60cctcccaagt ccctcgttat cttccgcctt ttccaccccc tccacacttt atttttccat
120ccatcatctt ccagtgcctc aatccaaatc actgtcgttg aatatagatt cgcatctgtt
180ttccgtacgt gtatttgttt actttcggga gggagattct cgccgttcat tccagatttc
240cgcccgtccg ccaaggcggc tgcgcctcca gccaagccaa aacctccttg gatcgccagg
300tggaaagaat ccctttcact cccccgtcga gtgagtctgt ctcactgctc cctccggtcg
360ttgcgcgcgc tctctctttc ctctccacac ctccagtata cttcctctgc gggtgtgtga
420gagactgcga cgatccggag aagaataaaa ccctccagtg cagcttaatc tccggagtga
480tccgtcatac atgaccggct cccgagccaa gatctcgact gggtccatcc agtgaaaggg
540aactcaatcg tttgcacttt tcgtcgctgc ctgatgctgt ggaccgaagg gctctgttga
600accctgactc tccggatctc tcggtgttgt tttgatgcca gcacgctcgt tccacactca
660cctttaacct ctcccatcgg ctgtcttcac tcgctgacag caatgcggtt tctcacttat
720tccctgccct tcattgcaag tgctatctcg ctctttgggg tcaatgtaca aggttagctc
780ggcgacatcc ggcgcataga tttgttctaa ctggatcaaa tacagctcga tcacaagctc
840caagtgccat ccgtcatgtg tcgacgcttg accagcccac catcaagaca ccctcacagc
900gggtcgatca ccttgaccac tttgacatca ccttcaatat tcatgacaag caccagcgga
960taaagctgga gctggagccc aaccatgaca tcctggcgga agacgcatcc gtacagtatc
1020tcgacgcgga cgggaacgtg cgacggcacg agcccattgc tccacatgag cataaggtct
1080tcaaggggag gagtctactc gggcgaggaa aaggcatgtg ggatccggtc ggatgggcgc
1140ggatctactt gaagcaggat ggctcagagc cactatttga gggagtcttc agtatcgacg
1200gcgacaacca tcacgttcag ctgaaatcgg catacatgga gaagaaacgc cccgtggatg
1260tcgaccttcc cgactcagcg actgactata tgatcttcta ccgggattcg gatatggtgc
1320gtctacatac ggaactcaag cggtcgtcgc tcggatcgac ctcgtgtcaa gccgatcagc
1380tcggcttcaa cactaacccc aaccaccctg tgctacaacc gtatggccag gcagagaccg
1440atacgtgggg agcaatttca ttgaactcct tgtttggact caacaagcgc caatccgata
1500tcggaagtgt gtctggcaat gcgggcggag tcaatctggc gtcgaccatt ggtgatactt
1560cgggctgtcc gagtacgaag caagtagctt tgattggtgt tgcaacggac tgcgccttta
1620ccggctcatt caacaacgag actgccgcca aggaatgggt catcagtact gtcaacagcg
1680cgtccaatgt ctacgaaaag tccttcaaca ttacgattgg gctgcggaat ctgactatca
1740ccgacagctc atgccccgac aacccgcccg cggccacggc atggaacatg ccctgctcca
1800gcggcaatct cacctcccga ctggatctgt tttccaagtg gcgcggtgag caatcggatg
1860acaatgctta ttggaccctg atgagcgatt gcgcgacggg caacgaggtc ggactgtcat
1920ggcttggcca actctgcaat agcgatgctt cttcggatgg ctcgagcacg gtcagtggaa
1980ctaacgtcgt cgttcggtct tccggctcgg attggcagat ctttgctcat gaatctggcc
2040acacctttgg cgctgtccac gactgtgact cccagacctg cgcggaggat ctcgaagcct
2100cgtcccagtg ctgtccgttg acctcgagca cctgcaacgc caacgggaaa tacatcatga
2160atcctacaac tggaacagac atcactgcgt tctcgcaatg cactatcgga aatatatgcg
2220cagccctggg ccgcaacagc gttaagtcca gttgtctctc cgccaaccgc gacgtcacca
2280cctacactgg cagccagtgc ggcaacggaa ttgtcgagtc cggcgaagac tgcgattgtg
2340gcggggaaga tggttgcggc gacaacaact gctgcgacgc gaagacatgc aagttcaagt
2400cgggagctgt gtgtgatgac tccaacgaca gctgctgttc aagctgccaa ttctcctcag
2460ctgggacggt atgtcgtgcc agtcgcggcg actgcgacgt ggcagagacc tgcagcggca
2520actccagtac ttgtcctacc gactcgttca agaaggacgg cacgagctgc ggcagcagtg
2580gctcgggact tgcctgcgct agtggccaat gcaccagccg cgactaccag tgccgcagtg
2640tgatgggcag tctcctccac agcaacgaca cctacgcctg ttcctccttc agttcctcct
2700gcgaactggt ctgcacctcc ccgaagatcg gcacgtgcta cagcgtcaac caaaacttcc
2760tcgacggcac tccctgcggt agtggcggct actgcagcaa cggcgactgc aagggccaaa
2820acgtcgaatc ctggatcaag aaccacaaag gtatcgtcat tggtgtcgcc tgcgccgtag
2880gcgccctgat ccttttggcc ctgatgacct gcatcgtaaa ccgctgtcgc cgggctcgcg
2940cgccaaaacc cgtcccgcgt ccagtgcctt acgggccgtg gcccggcgct aggcctcccc
3000cgccgccgcc catgaaccag tggccggcgc gaggctatca aggcttaggg aatgagccgc
3060cgcccccgta tccaggtgta cctggtcagc cagtaccgca acatatgcct ccccaggggc
3120ggtacgcttg attgacgaga cttcttctgt gcttctttcc atgcgatata gtatgcatta
3180cgattcttgc agcactagca ttacaaatgt gacgattact tcattgatcc tttattgacc
3240ttacattctt gtcttggagt ggaggcgtgt tctagagttt tggtttgatg ctgaattgtt
3300cttttctata cgggggtctg ggcgtttcgt gaattgcagt acttacaatc caggggtgca
3360atccccaggg ggttggttcg cgtcttcttt ccctttcttt gcccatatta gacggggccg
3420tgggatgtat ggtcggggtc tgatgtatgt attcctgccg tgtataatga acttttccgt
3480cataatatat tgcttcttgt
3500332520DNAAspergillus niger 33ttgatgcata atgctttcct tcagaggtaa
tacttcgagg acggctatga agcgtatgcc 60acatacgatt ggtatttgac gatatttgtc
atgctataca tgaaatttac atattgtata 120tagtgtcatg ggtttgaata aagacatatc
gatgtatctt aatagcaggt caattgatga 180cgctcttgtt gctccgacga aaagcgactg
catcgggccg atgcgggtcc agggtcgtcc 240ttatcaacgc cccgtcagca ctaacagttg
acatgtttct ccaggtcggc tagaaccgcc 300ttttcgcgag aaacgcgtgc tatcgcgagt
gcagggcaat tgttgcctag cagtagctaa 360agcctgagtt ccgtgatctt cacttctact
tcttcttctt ccctgctccc aacccattac 420tctgcacccc acgcccaaat gcgtttccta
agcagtgcag ccctattcgg cctggcgtat 480gcctccaccc aggcggtcct ccagccagag
gaaccatccg acttccgtac attccacagc 540ccatattccc cgcaccactc gatccgcatc
cgccagcaga atgaatcaat ctgcgctgcc 600cattccgccc aatacaccgg ctggctcgac
atcggccgta aacatctctt cttctggtac 660tttgagagcc agaatgaccc tgccaatgat
cccctcactc tctggatgac aggagggcca 720ggggggtcca gcatgatcgg tctgtttgaa
gaagtcgggc catgtctgat caatgagtac 780ggcaatggca cttactacaa tccgtggggc
tggtcccgga actcctccct actatttgtc 840gatcagccag tcgatgtggg attttcgtac
gtcgatgaag gagaggacct gccgggcgat 900tcgcatcaag ctgcaattga catgcatcgg
ttcttgcagt tgtttgtctc ggaggttttc 960ccgcaattgc agactcttcc cgttcatctt
tctggtgaat cgtatgctgt atgtatcttc 1020tctaggtcac ctgagctaag actaacttcc
acagggtcac tatgtccctt acctcggcag 1080tcagatcgtc caacagaaca agctctatcc
cactgagccc caggtccttc tgcactcatg 1140tctcgtaggc aacggctact attctcctcg
cgacactacc tacggctact gggaaaccct 1200ctgcaccact aaccctggag tccccgagcc
cgtcttcaac cgaaccagat gcgacatcat 1260ggcggccaat atgccgcgat gcatggaagt
atccgacgta tgtgttcgga accccgatcc 1320agctatctgc catgctgcgt cggaggtatg
ctacgagggc gtgatcggat ggtatgatga 1380cgagtctggt gaaggtggtc ggaataggtt
tgatagtgag ctttgcccca tccagttgtc 1440ccctgtcagc aaatctctgc aatttactga
tatgaaacag taaccgctcc ctgcgccctt 1500gacggcatat gctacatcga ggccgctcgc
atcgagcagt acctgaacac acccgcagtt 1560tgggctgctc tatcaccacc caaagaaatc
aaagaataca aggttacttc cgacaatgtg 1620tcgcgcgcat tcgatctcac ttcagacacg
atgacgccag cgtctgagca agtcgcgttc 1680ctgcttgcga atcaggtaca tttcctggcg
tatcagggca atctcgatct ggcgtgtaat 1740acggcgggta atctgcgctg ggcgcattct
ctgccatgga gaggtcaggt cgagttcgcg 1800tcgaaggcgc tgcggccatg gagttgggta
gatgtggtat ctggaaaagg tggagtggct 1860ggaacgacga aggaggtgag agtgaaggtt
agtgagagta cggataagga gtcgaggttt 1920gcgctagtta cggttgatgg ggcgggacat
tttgtgagta tccctttcgc cttgtgggat 1980ccaagtatgc ggattactaa tggtctatta
caacttttac agcttcctca agatagacct 2040gatatcgcgt tggatatgat ggtgcgctgg
atatccgggg catcgtttac tgagtgaagc 2100atgaatgtcg cagacgatgg gataccggca
tatttcacag gtttggtgta atatatgtct 2160tgtagctgtc ttcagcagta tttacaatat
gctgctgcct tctctgattt gggtataatt 2220atagtttaac acaatcaact gcatcggcag
agatggatgg gcttgatttc gcacccgcag 2280cagcgatgaa tttccgtcta accgagcgaa
cggtagcgga cactaacact gattgtattt 2340acaaccctgt ttctttcctc ttgatttgag
acagaagtat gtaagctagg cgtagtagaa 2400tacatttgtc atctacttct actgagttgc
ataacgtaag cgactattgg ccgctacctt 2460gccccagtta gttagttagt agtagtgggt
gccgaagtca gcacacattt ttcagccctc 2520341721DNAAspergillus niger
34attgatacga ggcgtttgta ggtggcgttc ctgaagagga tctgataccc actggttttc
60ccagtgggcc tatgttttcg tgatagtttc ggcatttcta tccatctagg aaaattttga
120tggcacctgc tagtcgatgt gcccttcctg atggagcgat ctcaaagtgt caaatttctc
180ggcgaaacgc gtctacccct tctttgggat aattccctgg tatgcagcat tcctgacacg
240agggattgtt tgggattccc cagattccac gtagcggttg gcaatgatgc ttatgatttt
300ataattctca tcttgtcctt tgtcctcggt gcagtagaga atcttcactg cccgatacca
360atatgacatt gttactcaac ttccacgcgc tctttacagt cattcttgtt gccaatcttt
420caaccagatg cagcgcactg ctctctggac gtgacttttg ctccacgcca gcgcccggtg
480agtcactccg agcggagcat aggaggctgt atgatgtaca ggcccaacgt gacagcaccg
540ccgaggagag ccgggaggtg gtgccttgga ttgaaatcga gacatggttt catattgtaa
600gcagcaatga agcagcaaac acagtatcag acgacatgat caccagccag gtccgtcacc
660attcaacttt tgacccatgc ctgcgcaatg tcgtgttttg gactaacaag tcctagcttt
720cctatcttca gaaggcatat gaaagtgcga ctatcaccta tcggttggag ggcataactc
780gtcacataaa tgactcgtgg gcgcgaaatg atgatgaact ggggatgaag aatgccctac
840gaaggggcaa ttatggcaca ttaaatgtct atttccaaac agatctccag gcgtcatccg
900acgagaattc tcgggactat ccaaatgacg gtaaccgacg aacagatgtg tcagatcaat
960catcatcaac tgtcctaggc ttctgtacgt tgcctgaccc gagtgtgaat tccagcagcc
1020ctcgttccag ctacatcaag gatggttgta acgtgttagc ggatatcatg ccgggtggta
1080gtttagcgca gtacaacaaa ggcggcacag cggttcatga ggttggccat tggaatgggc
1140tgctgcatac gttcgaaggt gaatcgtgct cccctgataa tgaaggagat tacattgatg
1200acaccccgga gcaatctgag cctacgagcg gatgtcccgc cgagaaagat tcatgccccg
1260atcttcctgg ccttgatgct attcataatt ttatggacta ttcatctgat gactgttatg
1320agagttttac tccagatcaa gcggagagaa tgaggagtat gtggtccgct atgcgggaag
1380ggaagtgacg gacggactcc aagggtatat aatttgcaca tataccagct atagatagac
1440aactatggta gcacgaatat tgatgtcaac atcttggtct ttcaattccg tgggaattga
1500ctatcacaat tcatgaacct tttgtagaat ttgtgtattg gagagcgatt gatgcaaact
1560agacactcgt tcgaaattca cagctagctg tcatgtggaa tgcgaaatca gatcatcgct
1620tgcgcttttt tccacgatct tcatcgtcca ggccactttc cctgtccttt ttctttgcct
1680ttgattcgag ttcgtcccag tcttcaccac tctcttcgtc g
1721353550DNAAspergillus niger 35cctatcctct gctccgcccg gtggagagga
tctattgacg ctatggttct tgagcaatgg 60cgacaagacc caccagtgga gataagagga
ccgaaactat ggcttgttca aggtctgctt 120cgataaaccc ctcatcacga taaaccaagt
tgggcttact aacaaggcca atccttcaac 180aatctcatcg cacaacgacc caaaatccag
caaggcacga tccaacatac cctgtaatga 240caggaaacaa attatcactc ctctgggtgc
tgatgcagac caactcagta ttccgcgtgg 300attgtccgac tccgaagaca agctttggtg
tctggtgctg atcgtccgtc gtggagccac 360cactagtgga ttgaattacc cacacaccca
ctacttatac tctttacgct gtctatcatc 420ttcgtgctga tcagggggtt attttcttgc
gctatttggt ttcccactat gcttctgttt 480tgcagggctg gtcataacaa tttggtagtt
tgggttgggt ggcttgttat ggtcgatgtc 540ggggaggggt tcaccgttga acaatatgct
tgtcgttagg taggttgaac tgatgcttgt 600gttttgtttc caggcgccct ggacagcctt
tggctatcag caatagctca atagctggtg 660ttttgctcgg acgcgggtct tggtcgggtt
gacccagtgt gatctacgct agacgctttc 720ctgattggcg atggatgtca cgcctcttat
cagggtttgg tgatcatatt tagcctacat 780aggcaggttc ccttccgctg ggtatggaag
tgcaaacgcc agcgtatgga gagtgaagtg 840atgctagtca atgggatcag cagcgttctt
gtcaactagc atatattccg acgtttccca 900ccgttcctga tgatggcttg catgccacgc
ttgctctgtc gctgcgatca acatgcatgt 960ctcacttttc ctactcagtg ttacggcagc
gtttgccagc ccaacacccc ataactatgt 1020tgttcatgag cggcgcgatg cattgcccag
tgtctgggta gaagaaagcc ggctggacaa 1080aggtgcccta ctgcctatgc ggatagggct
tactcagtcc aacctggatc gtggccatga 1140tttattgatg gaggtgtatg tgttccgact
cctcaccaga accaagctga ttgtccaggt 1200ctcatccaca atcgtctcgc tacggaaagc
atctctccag cgaggaggtg cacgacctat 1260ttgccccgtc gaatgaggcc gtcgagaccg
tccgaacctg gattgaatcc gccggaattg 1320ctccaagccg catctcgcaa tcatacaaca
agcagtggct acagttcgat gcccatgcaa 1380gcgaggttga gcagcttctg cagacggaat
actacatcta cacccatgcc gacacgggaa 1440gttcccatgt gacatgccac gagtgagtcc
attttctaca tgccatgaca ccgctaacca 1500gttaggtacc atgtgcccga aaccatccaa
tcgcacatcg actacataac accaggagta 1560aagatgctgg aagtgcgcgg cacgccctcc
aaaaagagag atgcagagaa gcgctctctt 1620ggcagtctgc ccccaatctt agcaccacta
ccaatcaata tcacgaagat tttcgacgac 1680ccgctagcac actgcgatct ggcggtaacc
ccagactgca ttcgaggtac gttcatccat 1740cccaattcaa gatttcacgc taaccgtcta
gccatgtaca acatcaccaa aggaacaaca 1800gccacaaagg gcaacgagct cggcatcttc
gaggacctag gagacatcta cagccaagat 1860gacctcaacc ttttcttcgc caactttgcc
aggttcgatt actctccacc ctcccttaga 1920cacaactaac accaccaagc gacatcccac
agggaaccca tccaaccctc gactccatcg 1980acggcgccac cgccccaaca gacgtcacca
acgccggccc cgaatccgac ctggacttcc 2040aaatcgccta cccaatcatc tggccccaga
acaccatcct ctaccaaacc gacgacccca 2100actacgaaga caactacaac ttcaaaggac
tcctcaacaa cttcctctac gccatcgacg 2160gctcctattg caacgaaacc tcctctctag
accctcaata cccagatccc tccccaggcg 2220gctactcctc ccccaagcaa tgcggcgtct
acacccccac aaacgtaatc tccatctcct 2280acggcagccc cgaagccgac ctccccatcg
cctaccaacg ccgccaatgc cacgagttca 2340tgaaactcgg ccttcagggc atcagcgtgg
tcgtcgcatc gggcgactcc ggcgtcgcct 2400ccagcacggg cacctgcttt ggcgatgcag
acaacgtctt cgtcccagat ttcccagcca 2460catgtcccta tctcaccgca gtaggaggca
catacctccc cctaggcgca gacgcagcca 2520aggaccagga aatagcagtc acccgcttcc
cctccggcgg cggcttcagc aatatctacg 2580cccgaccatc ctaccagaac cactccgtgg
agacctattt ctccactacc agcgacgacc 2640tcacctaccc ttactactcc ggagtaaact
acacagactt ctccaacaca gatggggtat 2700acaaccgcat cggacgagga taccccgatg
tttcagctat cgcagacaat atcatcatct 2760acaaccaggg cgaagcgaca ctggtgggtg
gtacgtctgc cgcggcgccg gcgttcgcgg 2820ccatgttgac gcgcattaac gaggagaggc
tggcgaaggg gaagtccacg gtggggtttg 2880tgaacccggt gctgtatgaa catcctgagg
cgtttaggga tgtgactgtt gggtcgaatc 2940ccgggtgtgg gactgatggg ttcccggttg
ctggggggtg ggatccggtg acggggttgg 3000ggacgccgcg gtttgaggat ttgatggata
tatttgtggg tgatgattga tggctgagac 3060aagatgtggg aaggtgtaat gaagatgatc
ctatttaatc cggtaatcta ccacacattc 3120atattactag tttatagcct agtcagatat
ataacattgt actactacta tagaggcgtg 3180tgatgcaaat gctaaaatat tgatatacta
ttaagcaact acattatgat ctaaacttta 3240cagttcatag tacagaaagg ctttgatcag
tcctagttca ataaaaggta tagactactc 3300actacatatt aaacaccaca actcgggatg
cttggggtat agtgatctga gagtcgcggg 3360aatgcccaaa tgctctgttc gatcatatct
tggttgatgt catgttggac gcattatatc 3420aaaagtcatc tagcgctttt ctgaaagttc
tattatgaga attttgcttg tttatagcta 3480gcgggtggat gattaatttt gtatgagtga
ttctatagac gttgatcatt tccatgattt 3540agaaactgag
3550362280DNAAspergillus niger
36ctgcctctca gcgaaaaccc caaaaccaaa catttctaga tcagacaatc aattgcggac
60tgagttccct gaatggtgga agatgccata ctgttagcgg ggactatttt tagcatgtca
120tgacttggtc ggcgtccgta catagaaggt tagcggatgt cttgatgatc acacaaaaaa
180gggataggat gaaactggat gtctcactgg ctatctcatt tgcccaacag catcagtcgt
240tccggcccgc tagttccacc acatgtcact tgtctcggat tattctcgaa ggtctggtat
300ttcaatgttc cacctggtcg ctctacataa atagaccttt acgccgtttt gttttcttct
360tcgttctacg tcgtttattt atccaaaatg gcttccaaga ccctcctact cattccggca
420ctggccacag ccgctctggg aagtgtattg gacctagata tcaaggttca aaatggctat
480gtatgtcaat gagaagctca gctccctgag aaatatctac taactcggcg aagaggacta
540tcgaggtaga ccttggaacc ccaggtggcc cgtttgattt gatgtacgac accggatcat
600caacactctg ggtgcttgat agcaattgta cagatgattg tccaaatgtt agcgggttag
660ttgattcatc ccaccattcc ttaagcctac actaatcacg agcgccaggt actcccgaca
720cggctacaac ctcacctcta ctggtgtcaa cttaggtgtc aacgacagca ttgcttacag
780cggaggcact gtcagcggct tcactgccac ggatattctc acggttcccg acaccaacgt
840ctcatatcgc cagagctttg ccgtcattac cgacagtacc tgggcggcct tagcagccga
900tgggttcatc ggcctggcat cgtctaccat cgcattcaag aatactacga cagccgtcga
960acagatgatg caggatggac ttttggatga acctcgattc gccatatatg caggttcagg
1020ggaatcgacc gtgaccaacc ctaatccgga gaataatggc gtgttcacct ttggtggcag
1080ccatgaggaa acctatgcgg acggggaact gcaatggatg aagatgctct ccccctttga
1140aatatacaaa acaaatctcc ttggaattca gggacacaac aactccgatg gccaggccct
1200gtcaagcgac gtcctgaact ggtacggcca ggttattttc gataccggta ggcattttct
1260accacactgt tttctttagc ttgctagact aatctattca acgtcgcagg tgcttcatcg
1320ataagcattc ccaacgacca gattgaggcg atgtatgccc taacgccttt ctcatacgct
1380gacatctcat ctggataccg acctctgtgc tccgatttca atgatacatg gtcgatctct
1440tttacaatgg gcttctatgg cgagggtgtc accttcaatt tgaccggtga tcagctggcc
1500gtgcctggct atcaggacga cgaccactgc ttccctccct tcaatccatg ggacagctac
1560aacacgatta ttggtcagca ttggttgagc aatttctatg ctgtattcga cttcggatca
1620ttcgacccgg agacatacga tatacgtgtt gggctggctc ctttgaagaa ggaatacctg
1680ccgagcgctt gacctaagaa cttctattcg tgtctttgag catttagttc aaacctcttg
1740ctcggacgtc aatgtttaag catgcgtaaa actatctact agaagttaat gttcatcgag
1800agttcatcgt ttaaataagc agcttatctt cattctttac agccactata cgcgacatga
1860caattgtttt atgggtcggt gcgataaagt ggccatctaa aggcaatagc aattaaaata
1920agccaggaga tggaacatga aagaatctta ctctttgtca tggcctcatg caactttggt
1980gcgggctgtc ggtgaccctg tggggcacat ctccggggag tggaggccgt taagcttcgg
2040acaacattat ttcttcagct caaccatgaa aaggttcggg gacctgcatg catcccccaa
2100gcccgcaagt ccgcgtcatc aacaaatatt gcgaagaaat ggtacgcttt agagccatgc
2160gggctatatt ttcggttcta tagattgtta gatataatta ttgcttgtcg aacccagatg
2220cagcattgat ctgttataat cctcatattc cgcgagtcac ttgattccgt gatcaagatt
2280372287DNAAspergillus niger 37tagttgaagg aggcttttgg ctttcttcct
ctttatcttt cttctgttgt cagtgattct 60gatcgcagaa gatgtgcaca agatgaagac
cgccccgagt catctatgga tactgaagag 120atggaagaga tctgacaacc aaggagtttg
tattatccag ttccagtttc atgttttgcc 180tccgtctgat agttccacca aagatactgc
cattttgtat ccgtagttag acggagacaa 240attgacaacg acagccttgg gggccaagtt
cagacatact tccagccgca cgcatggatg 300tataccttgt tgctacgtga gcgtacgcca
gcttatcgac ctattttagg acctgctccc 360acgtggagat tctgcagcaa acgcaggaaa
caaggaccag aaattcgtca tcatcgtttc 420tccttagcaa tcggctagga tgtttccctg
ctctcgtatt tggtctctgc tcgttgcagc 480cgccaccgct agtgctgtac ccaccagtct
ggccaccacg cacctgcaat cggttgactt 540gcttctgact cgcagttctt acgggtttct
tactgacata gcccttggaa ctccgggtca 600gagcctgccg tatctggttg actggacctg
gaccggccac tatgtggtga ccaccttgtg 660ctacaacgat cccaccgcca cctacgattg
tctcaacgtc gatcagaaaa ttttcaacca 720gactttgtca tccactttta tcaaccaaac
tgaccagtat ggctatcttt actgggatcc 780caaccacttc tactttacgg agcccgcagc
agccgatgtg gcgacggaca tgctgcgcat 840cggtcccacc gcggtgaaca ccaccatcca
agcagccaat ttcgtcttca acgagactat 900tagcgcattc cctttctcgg gagtatatgg
actctcacct gtttttcagg gtgacaatcg 960tgagttgttt ccccaccgtc ataaaactaa
ttcatgttaa cacgaatcat aggatccgtg 1020caagcgtcct tctaccaagg atggaggagc
ggcgcctggc actctccaat tgtctctttt 1080atctactgcc acgacaatgc caccaaagcg
gtatgcagtg gttacgacgg ccttcagaca 1140ctaggcggat acaacacctc tcacgtccag
ggagatatca cctggtacga catcattgtc 1200acggaggcga tcaacacgct ggactttgtc
tatgcgccag ccgtgattaa ttattgggcg 1260ttgaacctca cgcgcttctc tatcggagac
gaagagcaag agctcaacaa gaccactact 1320ctggatggaa agcaagccgc cgttgccgcg
ttcgaccacg cttcgtatgg tcgcggtgcc 1380ccagtgtctg tgtacggtta ccagcgtcta
gtcgagctgg tcggggcaaa agccgtcacg 1440ctttccgatc ctccaaataa cggtgagcag
ggattctatc agttcgattg ccggaactcg 1500agtttactgc caccgctgcg gtatgagttt
gccgggtcag agcgggcgtg ggagattgtg 1560cccgagaact atgtggaggt gctggcgaac
ggaaccaata agtgcacctt taatgtacgc 1620accctgggag atggagcgat ggtaatggga
aattttggcg agacatttgc cattgataag 1680tatgtcatgt ttgactttga gaagttgcag
gtggggattg cagacttcgc gtggtaatgg 1740tataaggatg tgtgtttgaa catgtgcatt
atctcgagtt tgactgaata gacggagaat 1800ccaagaccgt agatcatgat gctatctgct
tgattatttt caatgtctct agtatgggga 1860aagtaataca tgtgataata tctccaaagt
agtaattaaa agagaaaact aatcaccact 1920agtaattaat actgcaaata gcgctgttcc
acattgagac aaccatataa taataagtta 1980gtggatatac ggaatattag aggtcatctt
aataccatac tagtggatac aaccatccgt 2040acatacggaa taggaaggag gagcgagatg
gtcatcattt gacactgagc atactgtgtt 2100ggcccagtca agctgttgta tgggggagct
cctagtgtgc ccaaggctcc agcatcttca 2160ccttccgcaa cgactttact tagtttacat
taggtgagcc tttctgttac atagttgatt 2220gaaagcctct ccccaattgt cttagcatga
tagaggttac tcggagaacg tactaatcag 2280gaggcca
2287381950DNAAspergillus niger
38gatctttcta gagcattgtc ttgattgtgt cattctgtca ttgactccgg ctatgaaata
60ttattctcaa tctgcctaaa accaaattct actctatcac tacacatttg tatcacctga
120tctggctgag ataggagagt ccggcatctc atcgtctgca tcagacaatt gcgataaatt
180cattgcttgc acctgttatt gattcttcca agttatgcat ctcccacagc gtctcgttac
240agcagcgtgt ctttgcgcca gtgccacggc tttcatccca tacaccatca aactcgatac
300gtcggacgac atctcagccc gtgattcatt agctcgtcgt ttcctgccag taccaaaacc
360aagcgatgct ctagcagacg attccacctc atctgccagc gatgagtccc tgtcactgaa
420catcaaaagg attcccgttc gtcgtgacaa tgatttcaag attgtggtag cggaaactcc
480ctcttggtct aacaccgccg ctctcgatca agatggtagc gacatttcat acatctctgt
540cgtcaacatt gggtctgatg agaaatctat gtacatgttg ctcgacacag gcggctctga
600tacctgggtt ttcggttcca actgcacgtc cacaccctgc acgatgcaca ataccttcgg
660ttcggacgat tcttcgaccc ttgaaatgac atcggaagag tggagtgtgg gctatggaac
720tgggtctgtc agcggcttgc taggaaaaga caagctcacg attgcaaatg tcactgtacg
780catgactttc ggacttgctt ccaacgcatc ggataacttc gagtcgtacc caatggacgg
840cattctcggt ctcggtcgaa ccaacgatag ttcctacgac aacccaacat tcatggatgc
900cgttgcagaa agtaacgttt tcaagtcgaa tatcgttggc ttcgcccttt cacgtagccc
960cgccaaggat ggcacggtca gctttggcac tactgacaag gacaagtaca ccggcgatat
1020cacctacacc gataccgtcg gatcggacag ctattggcgc attcccgtgg acgatgtcta
1080tgttggcggc acttcatgcg atttctccaa caaatcagcc atcatcgata ccggaacttc
1140ttatgctatg ctgccttcaa gcgactcgaa gacgctgcac agtctcattc ccggcgccaa
1200atcttcgggg agctaccaca ttattccgtg caacacaact actaagctac aagtggcatt
1260ctctggtgtg aattacacca tctcgccgaa ggactacgtg ggagcaactt caggttctgg
1320atgcgtttcg aacattatca gctacgactt atttggtgat gacatctggc tcctgggtga
1380cacgtttctc aaaaatgtgt atgctgtgtt tgactacgat gagttacggg tcggatttgc
1440agagcgttcc tcgaacacca cctctgcgtc gaactctacg agctctggaa caagcagcac
1500ctcgggatcc actacaacgg gcagctcaac gactacgacg agctctgcta gctctagtag
1560ttcatctgat gctgaatcag gaagtagcat gaccattccc gctcctcagt atttcttctc
1620tgctctggcg attgcttcct tcatgctttg gctctagtta accgcatctt actcgacgcc
1680tgaacctcgg gaaacatatg cattatttac acatgctgct gatttgtatt tgcatatatt
1740cttcgagcct ggacggcgtg cgggtcatat taccttacat tcgaagtcct tctctaatca
1800atcaacattt attcttactc caccagttct ggctcgcaat taaccctgtc taagaaaaag
1860ttggtataga acatggcatc cactacctgg aacattcaaa gaaccttgtc cgggatcagt
1920gtgtatgact tcgggtacga ttctgacatg
1950392660DNAAspergillus niger 39accttctggg gtggattatc tgaagagatg
aactgccccg cccaagtgca agaagatgat 60gcctgacagt tgatgcctga gggatgcttg
tatcaggtga tcccggctat ggtggtctaa 120cccagttggg ctgaattgac cacatttagt
tagtgtcagc cttaaacaga tgaggctttt 180cgcatgattc atttccaagg aacccaagga
aatggtcaaa cagcaaagga ataatgaccc 240aacagttata cctagctatg cagcataaaa
gagaaaaaaa acttgaactg atacgagcca 300tcatgtaacc tccgtctctg gctcaattct
ccatccaatc acaattcttc caagcaaact 360tgggattcgc cgggttctcg gcaaccctaa
cctctaccag taaaggttga tccttccccc 420acccgatttc gcatggcttg gacccctgaa
atatagttcc tctacgtgcc tgaatatgga 480gcacgttatt cgtctttcag ggtcttgggg
ccttggcctt ggggctgcca agcgggcggc 540ggagtgtgcc ccgtggtgcc ctgttttgag
cttggatgcg attactatca tcacttaata 600agagcaagac cctgcgtcga tattgacggt
caatacactg acctcagaga accgtcgatg 660tgcctgattt cgctccatca aaatgacgtc
ttctaccttg cgccttgccg tcgcgttggc 720tttgtcaact tgcagcagtg ccctatcgag
ccagcgagat gattcacttg tggttccatt 780tccttttggc aatcttgagg atgtccatat
tgccaagcgg gatagctcca agacagtaga 840agctcctcta gtgatatatg tcagtcagct
ctcccactct ccatctatgt catcaactcg 900acaaccctaa caatcaacag ggcgacagct
actggatgaa cgcctcaatt ggaacccctg 960cgcagtcact aagtttccta ctagatctta
cgcgctcaag ggtcgagccc gcatacaccc 1020tcgatgagaa ttacgaatgt tctgacgatg
aactctgctc cgaattcggc ttctacaaac 1080ccaccgattc atccacttat cagcatctca
cctacacaca gagacacgat gcaggtgtcg 1140actactccta ccttgatacc ataactcttg
gagatcacgc aaccgacaat gtcccactgg 1200acatgtatct tttgtcctac atttcctgtc
agttccccct ctctccctta ccaccctatt 1260tatccatcat ctgaccattg tctaaccacc
acatagacag ctccctcggt ctctcctccg 1320tcaacaccag cttcccctac atcctggtcg
atcgcggcct caccacctcc ccatccttca 1380gcctaatcgg cgacaacgga aacaccacca
cccccagcat catctttgga ggcatcaaca 1440cctccaaatt caacgggccc ctgcaagcct
tctccttcgc agaccacagc atcaccaaca 1500atccattcgt caccgtcgaa gctgactccc
tccaactaac caccaacacc aacgataatt 1560ccacctaccc tattccctcc tccaccccca
tgatgctcag aaccgaagaa ctaatcacct 1620acctccccaa ctcgaccgtc caatccctct
acaccgacct taacataacc atggacggcg 1680tgatctccac ttcaagattc tacggggtcc
ttccctgcgc ccgccaggaa accgaatctc 1740acacaatctc tctagccatc ggcaacatga
ccttctctgt gtcctgggat gagctcttcg 1800tcccgtggac gcgtgacgga ctatgcaagt
tcggcattca ggcccaggat tcagattaca 1860aaactcgtgc ggagctgggt gttccctttc
tgagacggat gtatgtcgct gtggattata 1920ataatcagtt tgtgggcgtt gcgacgctga
aggatgatga tgatcagaat ggaggtgaag 1980atgagattgt ggagattggc actgggacgg
cgttgcctag tgctgtcggg gattggccgg 2040ctagtgttac ggcgtatacg cctgctgctt
ctacagggac ggcggctgcg acgttgacat 2100tcacgacggc gacgtctagc gggggaggtg
tggtgccgac gggtctatca gagttgggta 2160gggcgttttt ggtgccgggg gtgctgggga
tggctgtttt gcaggctgtt taggttagag 2220attgtggtac gattattctg taggttaaag
atgtatattg tctgttattg tacttgatag 2280cttgatatgt ataatgaaag tagtatagtt
tgaatttttg ttatacacca cgtgaaccca 2340ctggaccgtt gtctactgct cgcagtgagt
gatatagtgt agacccgtat tactagtcat 2400gtaggaccct accttgatac cataacactc
ggagaccacg caaccgacaa cgtcccactg 2460gacatgtacc ttttgtccta catttcctgt
cagccccttt ttgaatgtac ctacatccct 2520ctaaactcta gtactatcac gaacagcaaa
tgacattcaa tctattctat gcatattacc 2580cttctaacaa tacccagatt ctatcaaatc
acgtgaacat acagccctcc caagtcacct 2640gacaaagcgg acaacaatgc
2660402501DNAAspergillus niger
40tgattcccaa atcgcaccag atgaagttga atcagtggta cagtttagcg ggtggctgtc
60atgacagaca gaccctcaaa ttgaagcaat ggcagataga tatggtctcc agtccaccga
120ttgaatgcag ttggagaagt caatcccccc ttggaaagcc cccaacgttg cgctttctga
180ccacctgaaa tctgacaatc agcaatcctt cagctcttct cagcgtctct tggggccgat
240ccccatcgga tgaacacgcc tcatccctca ttaggcatgg ctgagctgca tgtcagcgga
300gtcccaacac ccaaaccaat cctttagcgc gcaccgggcc tgcgaagcca tcaccatcgc
360ccgccttgca gcaatgtgct ccccggtcgc attcctttcg tttcataatc tcacctccgg
420ccagcccata tccttcatct cgcagctttt ccctactcct tcttttcttt tgctttcttc
480ccccatcgca ggctgtttga atgcctgctg aagtatccat ggtatgatgc gaccgatact
540tctcccccta ctgggggtat ttctgcagac ctcctcggca tccaatccct atgtaatgag
600ctggtcttcc caagcctacg gtccagatgg cccgtggcag gccgtatcca tcgacgtggg
660cagcaaccag cagacggtcg atctttaccc cggagccaac tatgctagca cgatcctgat
720gagcactctc tgcacgaaca aaaccctgtc atccacctgc tacgctgccg aagcaggcac
780gttcaaccaa aacacctcca ccactgccta caccaccgcc agctcgtggg aaacaactta
840ctgggccgtc gagggtggaa gccaagaggc tgtgctcggc gatgaggtca ccttagggtc
900gtttgtcgtc cccaatgtga gcttcgaagc catctaccag acctaccaga cctatcccaa
960tggcatcgcc tatcctgtct cggtcggcag tctggccctg gggggtccgt acttgtcgga
1020taccgtctcc aattcgacgg tcctgaacat gatcgcagga tggctttact cgtccaacga
1080cattccgtcc tactcgtacg gcatgcatat cgggtcggta gaccccaaaa tcccaggctc
1140cctgatcttg gggggctacg ataagagccg agtgatcgga gacgtgagtg cgcagggagt
1200agtgtcttcg agtggtcttt tggaacttga attaaaggat attgggctgg gtgttgcggc
1260gggttcctct cccttcagct tcaacaacga aagtggcttg tttctccaaa gcagtggttc
1320ggttcaggcc aagaccgtcc agattgatcc aaccaagccc tacatgtacc ttccccaggc
1380gacatgcgat gccatcacct ccaccatgcc gatctccttc aattccagct tggggctata
1440cttctgggac accacgagcg atgattatct gaatatcacg tcttccgccg catacctctc
1500ctttgtgttc aacatgaatg gggtcaacaa caagaacatt accatcaaga ttcccttttc
1560ccagctcaat cttacgctgc aagaaccgct ggtcgatcaa aacgtcacct acttcccgtg
1620cttcctcact acctccaccc cggtgctcgg tcgagccttt ctccagtccg cattcgttgg
1680ggtgaactgg ttcaacggga acaactcggg cacatggttt ctggcacagg cccccggccc
1740gggttacgcc agtgaagaca tcacccggat cgcagtgagt gacacgtcgc tttctgcctc
1800taacggtacc tgggaagaga cctgggctac gtactggggc atcaaaacat ccgacaactc
1860gagcagctcc aagagtggcc tgtcttccgg tgccaaaatt ggaattggcg tcggggtggg
1920tgtcggtgga gcagtgttga tcgcagcagg tatagccatt gcattctgtc ttcgccgtcg
1980ccgcggggcg agtcaagagg cggctggaga gcaacggagg tcgatgttta ggggctttgc
2040ggagctaccg ggaggtgctc acagtgaacc ggcgaaggag ttggatacga agatgcataa
2100gccgccgcag gaaatgatgg cttcgcagga ggtagagcga tacgagctgg ggtgatggct
2160aattctaaat tttaatatgc gaagggatcc ctacaggata ctacaggata ccatatacgt
2220gcgaatataa tagctagttg ttccaatatt atgtatcaag cttatactgt ggattaacca
2280acgacttgtc tccgatacat agggtagtac gtagtcatgg atccggggtg tattgtcatt
2340cagcatgacg cattgctcaa gccaaaagta agctgggcca gtgtatcacc catgaaagca
2400accctctctc acctctgtac tgatgacttg gtggaaattt ggatcctcgg aaattgtgat
2460taacacgtgg atctagaccc gaccaggcgg gagcttggca t
2501413570DNAAspergillus niger 41gaaaaggatt gtccccatgc ataataggcg
ttccaccacc tgcatatcca tacatgcagc 60ccgagttatg cgatgtgatc gggcctggaa
taacatgatc tccgatctaa cgggggccca 120aaaggggcag ccaagatctg ctaggcccaa
ccaggctagc ggcgggccat ctgaggcggt 180gatccatttt gggcgaggaa aaggcaaaaa
gtggatcggc gaaaggcaag ttggagtgtc 240gggagtttgg tctcccgccc cagggccgcg
tattagtcta gctctctttc tattcactcg 300ggccaactcg tcaactctct ccctcgcttt
cattcagatt tcattatttc tcttacggtc 360ttgccattct tgccactttc ggtccttatt
attgcttttg atccccagtt cccttttccc 420aactgggtgg cgctcctaca gaccgacaca
tacctggcgg actttccttg cttctggctg 480tacaccggga cccgcctcat acccagtacg
tgttggtcca tggcaagcct cctacttggc 540ctgattacat cgtcctgaga gagagagttc
accaaaactc tcccccaaac gatgcgtctt 600acaggtggtg tcgctgcggc tctgggcctc
tgcgctgctg cctccgcttc tctccatccc 660catcgttcct acgagaccca tgattacttc
gctctacacc ttgatgaatc cacctcgccg 720gccgacgtcg cccaacgact aggtgctcgc
cacgaaggcc ccgtcggaga attaccctca 780catcatacct tctcgatacc ccgtgaaaac
agtgacgatg tccatgcgct gctggatcaa 840ttgcgcgatc gtcggaggtt acgccgccgc
tccggagatg acgccgctgt ccttccctcc 900ttggtcgggc gagacgaagg tctaggtggc
attctttggt ccgagaagct ggctccccag 960agaaagctcc ataaaagagt gccgccgaca
ggatatgctg ccagatcgcc cgtcaacact 1020cagaatgacc cccaagcgct tgcggcgcag
aaacgcattg cctcggaatt gggcatcgcg 1080gaccccatct tcggcgaaca atggcatttg
tataatactg ttcagttggg ccatgatctt 1140aacgtgacgg gtatctggct ggagggcgtt
acagggcagg gtgtcacgac ggctattgtc 1200gatgacggtt tggacatgta cagcaacgat
cttaggccga actattttgc ggcgggttct 1260tatgactata acgacaaagt accagagccg
aggccgcgct tgagcgatga ccgccacggt 1320actagatgcg cgggtgaaat cggtgcggcg
aagaacgacg tgtgcggggt tggtgttgcg 1380tatgatagtc gcatcgctgg tattcggatt
ctctccgcac ccattgatga cactgatgag 1440gctgcggcta ttaactacgc ctatcaggag
aacgatatct actcgtgttc ctggggtccc 1500tatgacgatg gcgccacaat ggaagccccg
ggcaccctga tcaagcgggc catggtcaat 1560ggtatccaaa atggtcgtgg tggaaaaggc
tcggtttttg tgtttgcggc tggtaacggt 1620gccattcatg acgataactg taactttgac
ggttacacca acagtatcta cagcatcacg 1680gtgggtgcca ttgatcggga gggtaaccat
cctccgtatt cggaatcctg ctcggcgcaa 1740ctggtggttg cctacagcag cggcgccagt
gatgcaattc ataccacgga cgtcggcaca 1800gacaagtgct cgactaccca tggtggaact
tcggcggccg gcccgctcgc tgcgggaacc 1860gtggcgctgg ccctcagtgt gcggccggaa
ctcacctggc gtgacgttca gtatttgatg 1920attgaggcgg cagtgcctgt tcatgaagat
gatggaagct ggcaggacac taagaacggg 1980aagaagttca gccatgactg gggatatggt
aaggtcgaca catatacgct ggtgaaacgg 2040gcagagacct gggatctggt gaagcctcaa
gcctggctcc attccccctg gcagcgggtt 2100gagcatgaga tcccacaggg cgagcagggc
ttggctagtt cgtacgaggt gacggaggat 2160atgttgaagg gagccaacct ggaacggctg
gagcatgtca cggtcaccat gaatgttaac 2220cacacccgcc gaggcgatct cagcgtggag
ttacggagcc ctgacggtcg ggtcagtcac 2280ctcagtacgc cccggcggcc agataatcaa
gaggtgggct atgttgactg gaccttcatg 2340agcgttgctc actggtaagt aaactttttc
tcggttgtcg gttcttctgc taatacatat 2400ctaggggcga gtccgggatt ggcaaatgga
ctgtgattgt caaggacacc aatgtcaacg 2460agcatactgg gcaattcatc gattggcgac
tcaacttgtg gggcgaggcg attgacggag 2520ccgagcagcc tctccacccc atgcctactg
aacacgatga cgaccacagc tatgaggaag 2580gaaacgtggc taccacgagc atcagcgccg
ttcccacgaa aaccgagctg cctgacaagc 2640ccactggtgg cgttgatcgc ccggtgaacg
ttaagcctac aacatccgcg atgccgaccg 2700gtagtcttac agagcccatc gatgatgaag
aactccagaa gacccctagt acagaggcaa 2760gctcaacacc aagtccttct ccgaccaccg
cgtcagatag tatcctgcct tccttcttcc 2820ccacgttcgg tgcgtcgaag cggacgcaag
tttggatcta cgctgcgatc ggctccatca 2880ttgtgttctg cattggcctg ggcgtctact
tccatgtgca gcgccgcaaa cgtattcgcg 2940acgacagccg ggatgactac gatttcgaga
tgatcgagga cgaggatgag ctacaggcaa 3000tgaacggacg gtcgaaccgt tcacgtcgcc
ggggtggcga gctgtacaat gcttttgcgg 3060gcgagagcga tgaggaaccg ttattcagtg
atgaggatga tgaaccgtat cgggatcggg 3120ggatcagcgg cgaacaagaa cgggagggcg
cagatggaga gcattctcgg agatgaagtg 3180cagtagatga gggttgattt tatttcggac
agtgtttctt acttgttgga tgacctgcgt 3240tgaacaatat tcctgctgtg tatgctgcat
agagaaagcg tgtatatacc atgtatgtgt 3300gcatcatctt tgatcgggtt attattcttc
atctgccatg gtttgtgatc tccggaatag 3360taccaaagga acactaaatt aagggtcttg
gcgatgacgc ttcccgtcgc tgcttttgac 3420ttcctccgca tctcgtctct cctgctgttg
accgcccgcc aaccaacctc catctcctca 3480ctcctcccac cttaatcttg ctgtcctgct
tctagagccc cccagtttaa tttaaaaacc 3540ggcttttcct agctccacgt attgtacctc
3570421236DNAAspergillus niger
42aggtcccgga cagatgacaa ggctctaaga ctgtatgtct cattcagggt ctagccgtgc
60ccgctcttat tggcccggac gcttcacgcc gcgattactg cgcccccgca tgtctctctc
120tccgctcgct atcgatgcgg tgtttgcccg ccattgatgg gaggaatgtc tcacttcggg
180cttgtgcttc gcagatccat acccctcaat gacctagtat agcaaacagg cgtgatgtca
240atgattgtgt ttctattcag agcatcaggc ctcccatgtc tatacctcag ttatggagcc
300ttggccgcct tgggtaggca aggcctcccg ggtattctca ttgccatttc accgctactg
360cagcttcaag tgaagagaaa gaggggaaag aaagaatata agtagcttga caatctcctg
420cacgtcctcc tcttcagctc acatcaaaca tctattcttc actcctcaag agtaaaccac
480tactaccaaa cacaatctca tctatagctc acgagtacca gcatcactca tctaccacaa
540tgaagacctt ctctaccgtc acctctctcc tcgctctctt ctcctcggct ctggccgcac
600ccgttgacag cgctgaagcc gccggcacca ccgtctctgt ctcatacgac actgcctacg
660atgtctctgg agcttccttg accaccgtct cctgctcgga cggtgccaac ggcctgatca
720ataagggcta ctccaacttc ggctcccttc cgggcttccc caagattgga ggcgccccta
780ccattgcagg ctggaactct cccaactgcg gcaagtgcta cgccctgacg tacaacggcc
840agacagtcaa cattctggcc attgattccg cacctggtgg cttcaacatc gctctggagg
900ccatgaacac cctcaccaac aaccaggccc agcagctggg tcgtatcgaa gctacctata
960ctgaggtgga tgtcagtctt tgcgcataaa cgccaaacat tcagcaaatg cttgggggat
1020gaattcggcc atggcaggga atggttatag ttggatatga tttcgggatt tacgattgat
1080acgcttattt gggatacttt cctttatctt acctattgtt aatagcataa tacggagcat
1140cacatacgta catacacata catcaatata gtgatatatg aacttgaata cagagctttt
1200tgaaaatgaa atgagagatt ctaaggtcgt atctac
1236431750DNAAspergillus niger 43gtgaaagata tcggatagcc gtagaccctg
aaagattatt tgagcctaag ggctcttgca 60gtgacgctag caacagcgaa cccactggga
tccaaaacac tagatgaagc tattgtaagc 120cggaaccctt ccggcgcccc acccagctcc
tgcaggggcg aaaacatgac tctctaactt 180aggagaaatc acagccaagg cccaagttca
ctgagagacc gaaatggcgt gaatttaaat 240gttgcctttc gcgcacttta ctattccctt
gttcctatca gcagtcactg cttggcctcc 300tttctagtga gcagttccat tcactttagg
ccccacggct tcgttacaag agcagtatgg 360ctcaaatatt ctggctttca ctcttcctgc
ttgtctcttg ggtcagagcc gagtccaacc 420gcaccgaggt ggacctgatt ttcccaagaa
atgatacctt tgcgccaatg cctttgatgc 480cggttgtatt cgccgttcaa gccccttccg
tcgcccataa agttaataca tacatcgagt 540acggctatta cccagtaggc cgtccaaatg
aaacagttat tggccagacc gaccatgtgt 600ccgactcaac aaacgaaacc acttatttca
gtgtctctgg tatcggcaga acgttcaata 660ccactggcag ctgggagctg ttttggaggc
tgagatggac caattgttca atctcagaag 720actcgagata ctacaaccaa tcctacccct
ggatatcctc cccatacatc gacggtagcc 780tcaacatcga caaggtctat gagggctttc
actacacagc atacaatgtc attgtcgaca 840gggttacctt cagcactcgc gaagatgcta
gccaacccaa cctcacgacc ctcaccaata 900gcgagaactg cgataaagtc tcgtctcttg
ctctattgtc gattgtggac tccctaagga 960ttccacccca gttaccccaa gaagatattg
ataccgtgtc aatgtgccca caactcgccg 1020atgccaggct aaattcaact tcaacttcaa
gcccctgcag cgttagcatt agtcccgagg 1080ttgagtctaa tatcctggcc aagatcgcag
acaatgaatg caataacgca cttcaccccg 1140ctgtgagttg caccactgaa gaaaccaagg
aaggcagcgc gagcagccat gaccacggcc 1200atgctgtatg gcttgtcatt acgctagctt
ttgccttcct tttctaaagg atccaatgct 1260agatgtgctc gactcaggct ttgagacagg
tagacaggaa aacaggcttt attgtcacgt 1320cgccagtagg ggaccatcaa ccacacatat
gtccctacga gaccatagct atcaggaaca 1380atgtacgggc tggaactcca ctcactctta
ggttgtaaat cccatttctc aatacatagt 1440ttgtgagcaa accacgcttt tcttgaatca
ttgcatcctc tttagcttca attccttacc 1500cacaaacacg agaggagcgg acccgccatt
ccacatagtg tcttggcagg gtcgcctctc 1560caactacaat gcccaacttc agaggcccag
actatctctg ccaagtgcca aggctggact 1620caaattacgg aactctcgga tgcttgtgcc
atggcttatt gcttgaaggg tttggaggtt 1680tcggtggtta gcgcccggat aggcagtaga
acacctccaa atgaggaatg cggccgaagg 1740taggtggggc
1750442030DNAAspergillus niger
44tggatgccga gcaaattaat taactgcgct tacatggctg tcctaacaaa tcccttggca
60ctcttagcag tggtgcctgg atgggacgaa gccctgggca tagcttgttc atcggtaggc
120cgaaactctc ttttacgtac cgtacggcca atatcagtgt gaacccgcaa tttagatact
180attaacttac aagtaccctc acgttatgta gccgaatgtt taaaagcgat ggaccggttt
240aaagtctccg ttgtgcgatg gctgttatta acttggatta acttgcaatt tgcatcaagt
300aagacgtgct tctttttgga tctttttaca cacattactt acggcggatg ctctcgacac
360tccggatttc accagggtgg tgttagtcca ccatcggact ccgatgcatc ttaacatggg
420tggtcgagat gtcgccattc tcagcaggca ctttgctgtg acatcctcac aaagtgttaa
480tggcgttgtc tctgggatgg tatgaggaag aatgatgcca tccgtgaaga gtgtcgactt
540gatagatact caacaaaaag tcatcctata agtattgggt tggatctgct gttcttatca
600tttcgtctcc atcccactgc aatcttcttc aagacttcaa agtccatagt tccaacacac
660agtcacctct tcacccagct tcactaccaa ccaattcttc aagaagaagt tcactgctgc
720aattgctact gccattttcg caagcgttgc cgtcgcagct ccccagcgtg gcctcgaggc
780ccgcctcaag gcccgcggca gcagcaaggg atcccgaccc ctccaggcag ttgctagacc
840tgcatcaacc aagaaccaga ccaacgttga gtacagctcc aactggtccg gtgccgtgct
900ggtggagcct ccctctgctg cagcgaccta cactgcggtg accggcacct tcactgtccc
960tgagcccacc ggcaactctg gaggcagtca ggctgcatct gcctgggttg gtatcgacgg
1020tgatacctat ggaaacgcca ttcttcagac cggtgttgac ttcaccgtga ccgacggaga
1080ggcctcgttc gatgcctggt atgagtggta cccggattac gcctacgact tcagcggcat
1140cgacatctcg gcaggcgatg agattgttgc cattgtggag tcctacacct cgactaccgg
1200tattgccatt attgagaaca agagcaccgg ccagaaggtg tccaaggagc tgtcgtccag
1260ctccagcctc ggtggacaga acgctgagtg gattgtggaa ggtaatatga aacatacatt
1320cctcattcaa cattaccaaa cgctaacgcg atcttagact tcgaggaaaa tggttcgctc
1380gtcaacctgg tggactttgg caccgtcacc ttcactggtg ctgttgccaa ggcggcgggt
1440ggtgagagtg ttggacttac cgatgcgacc atcatcgaga ttgaggagaa tggccaggtt
1500gtcactgacg ttaccatcga cagcgactct gaggtgacca tcacctacga gtaaatttgc
1560acacgaggcc tgtcactgtg actgacggtg ttgagtgtct caatacaaag cgggttgatt
1620ggtatgcagg gatgtgatga tgtgatcagg tcagcgcttg atttcaatcg acgaggcagt
1680ggagaacgaa gatgtagata gttttgattt ctagtactac tttgcggggc ccctgttgaa
1740tttgagtcat tcttttgatt gatcctccgg aatcatatat caagtaatgg tgcagcgtag
1800ggagtactcc ataaggcaga tgaagtacag gtcttaaaac ggtatgatcc agtaagacaa
1860agcggatctt gtattccaaa caatcacgtc ggtaacatgg actactactt tcatctataa
1920tgtttgtatg ccagttgtgc tgcagattat taatttgtat aggcattgta caggtgcgct
1980acattgttca gcaagtaaac gtgattttga agtccccaag aacacaaagc
2030453080DNAAspergillus niger 45cttcggctgc gtcatgattc atgcgtcaaa
tcagtggact cttagtaggt ggatgtacag 60cacttatttg atacttcgag tgtccgccaa
catccggccg tctatattct ataccaatcc 120acattatgcc caacgaaact gtatgctgtg
atttaagcag aagtgagcaa agcactgtgt 180cagctgatct aatatactat atgctccgat
taaacccaag cagtctgcta ccagggagac 240taccaggcag aaagtatctg gctgcatttg
tcccagtctg cacaggactc gctatgttat 300ggagcagaag tattgtgcat ggattccatc
tgtcccacta gcatgactat ctcaatatcc 360accgggtaca tgatttagcc ggtatcactc
ccccgtccct gtagcaaagg cccgcgcaag 420aggtgtttcc agaaagggct ccacgcttag
caccgttcag caaattctgc atcagccgcg 480gggaaactgc ggtttgttgc caggtcgagg
tgcttaaata ccaccgtctt ctcgctcgtc 540agtgctctgc tcaggcgtcg ctcagtcaag
ttcagttcaa gactcaggat atccgggcag 600tcctccgcaa ctatgcgctg ctccctcatc
tcccttctag gcctggcggc catcccggcc 660cttggaggct gtcccttcgc acacactgcg
aacatgggca ttgataacat ggtgaaagca 720cacgctcaca tgtcccgacc gttgattgcc
tccaagagca gcccctcaac tgttcctacc 780tcctctagca ccccttctgt cgggcagaaa
ggcgtgttca tgatgaaccg cattgctcct 840ggcacatccg agctctacat tgccaacaca
gatggcagta atgaacgccc actcctctcc 900aaccccgtct acgagtacca tgcctccttc
tccccggatg tagaatggat caccttcacc 960agcgagcgca atggtgacgg taactctgac
atctaccgcg tacggaccaa cggctccgat 1020ctccaggaat tggttgccac gcctgcagtg
gaagactccg ttgttatctc tcccaacggc 1080cgcctggcag cctacgtctc caccgccaac
aacatgaagg caaacatctg gatccttgat 1140cttcagaccg gcgcgcagtg gaacctcaca
aatacaccca ccactgccgc caactcctcc 1200ctcatggaga gctatctccg tcctgcctgg
tctcctgatg gcgaatggat cgccttctct 1260tcggaccgca acacccaatg ggacggacac
ggcgtaccga ccttcctcgg ccgcacgggc 1320tgggagacga cgcaagaact ctctctctac
gccatccgtc ccaatggctc tgacttccgt 1380cagatcatct ccaagccata ctactctctt
ggatctccga aatggtcagc agacggtaaa 1440cgcatcgtct actacgaaat gacccgggaa
gacacctaca acgcccatcg tccagaaacc 1500attaccacag ccaactcgac gatcatgtcc
gtagacttcg agacaggcac cgatgtgcgc 1560gtggaagtcg ccggctccgg tgtcaagcaa
ttccctcagt acctggacaa gaacggcacc 1620atcgcctaca ccctcaaagg cggcaccagc
gagggcttct acacgaccgc gggactctac 1680gtcaacacga cctcggcgac cctcaggtcc
ccggcgtggt ctcccgacgg caagcaagta 1740gtctacgaaa agagcacctg gagcatccgc
tcggggtaca agcagctcta cagctgggac 1800agtgactggg actaccgctt cacggacgtc
ttccctcagg tctcgcacca ggagcgcgtc 1860gccatcacac agaagcagct gggcaattcg
tccatcgtga cgttgaacac aaccggaggc 1920gacttgcaac tcgtctacga ccccagcacg
gcggactttg tcagcgatga cgaaaccaca 1980ggactgagcg cttaccagcc cagctggtca
ccctgcggcg agtggctcgt cttcggcgtc 2040ggattctggt tcgagacgag agaagcctca
ggcggatgga tcgtgcgggc caccgccaac 2100gggagctact cggaggttct cgtgaacagc
agctactcca tcaccgagga tggagccctg 2160aacagcgggt tcccgagttt ctcgccggat
ggcaagaaag tggtgtatcg ggtttgggga 2220gccgacactg caacctacgg caacgccagc
gagatcgggc tgcgggtgct ggacctcgag 2280acgcgaaaga caaccgtcct aaccacagaa
tgggacaatc tgccccagtt ctctcccgat 2340ggagagctca tcctattcac acgcaaaacc
agcacgtaca attacgatgt gtgcacgatc 2400cggccggatg ggacagatct ccgcgtgttg
acgagcagcg gtgctaatga tgcgcatgcg 2460gtctggtcgc aggatggacg gattatgtgg
tctaccggca tgtatgggtt ccggtttgag 2520tgtgcgctgt atggtgatac gttccagccg
tatgggcagg ttatgattat ggatgcggat 2580gggggaaata agaagttgat gaccaactcg
atgtgggaag attcgatgcc gttgttcttg 2640ccgagggagg tactttagtt ggtggcggga
ggatgctctg atttttagag gatatgtgac 2700gtttgatgta tgataagaat tacacaagta
gttttgatca taacgcgttg aagaggatga 2760aaaaaaaccc ccgttgttcg acttgatctt
ggcgatccgg tgtagggttg ttggacgagc 2820aggatgtact gcttaattca agcaagtttg
ctactccgat tcagtatgga ggggcaatta 2880atgatgacga tattctgcac tttctactct
acctactact tatctgtata tgaatgagaa 2940aaaaatagta tgcaacgaat atagatcgta
ttgacagaac ctgtcagaga gaatgacctc 3000gatagtcgac tccatagcat ttgatctttt
actaacagcg aaagattccg gactaactat 3060ccatatatag tactcctaca
3080463290DNAAspergillus niger
46ttccaacttc ccatgaagtg atgtagtagt ctgctaataa tacctcaaga tagaatatag
60tatttactac ttatcgtcag ctttgtcgaa aattcaaagg tttgtggaag gccacctcat
120ccgcacctgt tgcctacagg gatagcatcc tgcgaggtgc ttatccgctt atcagcaaag
180gtggcagggg gggtaaggct atcccgggtt aggaaggccc gcggcacgat gcgatccttc
240cagactagga agctacctaa tagggacagt aagggaccaa tctcaggact aaaccgtgca
300gcctactgaa tctcaccact cggacctgat atttgccttc ggcttgcagg ggtatatcat
360ctcaatatcc tctagtggac ggaaactagc aactagtact ttgagctgac tcggccacta
420taaatatgcg gtcttatgct cctcaacgac aggcaactgc tctgactacc aaaatctgag
480taggactagc accgttgcga gctcctcttc acgataccga atcttttata agcaagtggt
540ccaatatcca ggtctaacgt atccatacaa tgcctccgga tgcaaaatcg cctggctacc
600agcctggtat ggcagtatta ccatctaggc cacatcctgc caagggaaaa gccattcgat
660tcctcctttc ccttgcattg gtcgcgtttg ctattgttca attatgtggt aatttccaca
720aaaataggag cgttgaacaa cagcttcaga gtcaaacact tgatgatgag tcctttaaat
780gggaagatgt gtgtaaatct atcacgttgg gatgtaacgt tctaacgcac tctattaggt
840tactcctacc aagcaactcg tataccatcc atgctttggt gatcacgaat gcgctcgctt
900gtcgcttcca atgaattgga accgaactga tggtgaaggg tcaaaaattg ccttggcggt
960tatcaaactt cctgccaagg tacctgtcac agatgcgcga tatggtggtg ccattcttct
1020gaatccaggt atgtagagcc atattgctac tcttcagcgg ctgtgctgag tttcaaatag
1080gtggtcctgg tggatccgga gtgagcatgg tctttagata cgggaaagct atccagacca
1140tcgtcgactc cccagaatca ccaagtgcag attcagcgag cggaaagtat ttcgatgttg
1200ttagctttga tccaagaggg gtcaacaaca caacacctaa tttttcctgc ttccctgacc
1260ccgcgacgag gaaagcgtgg ttactgcagt cagaggcaga gggtctactt gggagttctg
1320aaggagtctt cgatactcga tgggcaaggt acgaagcttt tggtatgagc tgcaatcaac
1380aaggagtcac agcgtcaaag gatggagaat ggataggaaa attcatgaat acggcccccg
1440tggtggcgga tatggttgaa ctcgtagagc gccatggaga gtggcgtgaa cgggaaacag
1500agcggctact ttcgacagct ccgaacactt tcccagttgg aacaaacgtt gacgccgaga
1560ggataaggct gcacaaccgt tggaaaaaag gggaggagaa gctgctatac tggggctttt
1620cctatgggac aatcctgggt tccacgtttg cggctatgca gcctcatcgc ataaaccgtg
1680ctgtcataga cggagtctgc aacgctgatg attattacgc cggcaactgg cttaccaatt
1740tacaagattc ggatgcagca ttcaataaat ttttcgagta ctgctacaca gctggcccat
1800cagcgtgtcc gtttgcgctc ggcggagatc ccgaagatct caagtctcgt tatgagcaga
1860ttttgaccaa tcttacatcg agccctattg ctgtgtctcc ttctggaaat aggggcccag
1920agataataac ctatagtgat gtgaagtcat tggtcgtgca agctctctat gtgcctttga
1980aattattcga tttggtggct aggctattag ctgagctcga gcaaggtaac ggctcttcat
2040tcgctgactt gaagtatgaa gccaaacaat ggccagtacc gcctccatgc gattcctcgt
2100ccacacaata caaagtacct ggcgagagtg atcaggaggc cgggaggaat atcctatgta
2160cagatggtcc aggcctcgac ggaactgcca aggaggattt ccggagctac tggaatatgc
2220tccggggaca aagtaaggcg gttggagatt tctgggccga ggttcgcatg tcgtgtgtca
2280aactggagac gcgacctgag tggcgctatg atggtaggtc cgaccaacac catattatac
2340acacaatata agctgacagg tatgcgtatc caagggccct tcgcaggcaa tacatcgcac
2400ccattgctgt ttatcgggaa tacttatgat ccagtaacgc cgctacggaa gtaagctttg
2460ttcacctgat cgtagcagac tttgaagtac agactgacgc ataggcagtg ctcatacgat
2520ggcgcgtgga tttcctgagt caatcgttct agagcagaac tctgtcggag tgagtacctt
2580atctgttccc caacccatat gcaagatgac taactgttca atgtcgcagc attgcacact
2640gagtggccca tccttgtgta cagcgaaagc gatacgccag tatttccaga ccggagagtt
2700acctgacccc ggaactgttt gccaggtaga ggagcttccc tttcgtcttg ccggatatga
2760gagaagtcag gtcatgtcgc caggtgacac agaattgatg tccgccttgc attcgctgag
2820cgagttccgc catctgctag gcgcgtgaaa aaggttaaat aagtgtcgcc agcgagcgct
2880gcagttatta atcctttaca tcataggagc tggctagaat gttaacgatg cagtttgccc
2940ctgtatctcg accaagagca tgaataacac attatatcag caaatgttat tgagcaaatc
3000aattattcac ctgcactcaa ttcaatttaa gatctgttgc ctagactacc agaattatta
3060aggcgacctt atcgaagcat ttgcagacca acccctgtcc atcgtcaggg gcaagacaga
3120catttcttct agaacccgga tgatgtataa tcgcggtggg gaagatcatc ctaaaccatt
3180gccccagact cgcctacctt aggtaatcca tacaaatgcc ggggaaatca ctgaggtgtt
3240gttcatcacg gcgttgaatc ggcttctgcg gtattaccac cataactgta
3290473080DNAAspergillus niger 47gaaaatcccg accgttaggc tacctaatct
gctgtcttta cccccgttag cccacttacc 60cagctttagt ctctcgaagg aaaagaaaac
aaagtaatcg ggagcctata atattcggcc 120agaagtgggg caatcacttc aaccgttcat
ctgttattga gttatcaagt ctgtcaagtt 180cgtcggagct ggataaggta gtcgccatcc
ccatttccgc tgtgcaatgc cgcagcgccg 240aagtgtctgc agctagctct attttgttca
gctacgtatc tgtctcttaa ccaaatggat 300gtcgccattc gcccactaac aacgccattc
cccacggccg acggactccg gcgcaagggc 360ctgataagcg ttagctccga ctccgagtta
gtcatgcggc ttattacgcg cccttctccc 420ctcacgatgt tgatgctttt tctggtatct
ctggcataac tggcaaagat caggctgtgg 480tcataagagt atcattgcct tcagcttctt
cgccatgttg agtagtctgc tgcttggggg 540tcttctgggt ctagcgaccg ctcaatttcc
tcccgagccg gaaggcatca ctgtgctcaa 600gtccaagttg catgagaatg tgactatttc
tttcaaagag gtgtgtgcag agtatctaga 660aatagctttt atgctcgatg ccgtgctgat
tgtcagcctg gaatttgcga aactacgccg 720ggtgtccgat cttattcggg ctatgtacac
cttccccccg gtttcctttc cgacgggaca 780ggagaagtgc aggattatcc tatcaacacg
taagccaatc tcgaaacatt ggaggatgag 840caattactga gcctcaacca gcttcttttg
gtttttcgaa gcccgcaaag atcccagcaa 900tgcgcctctg gccatctggc tcaatggcgg
tccgggtggc tcgtcgctca tggggctcct 960tgaagaatta ggtccttgtt ccattgcatc
agactccaag accacagtcc tcaatccttg 1020gagttggaac aatgaagtca atcttctatt
ccttgaccag ccaactcaag tcggcttctc 1080atacgatgtc ccaacaaatg gcactttggt
tcggactgcg gacggcgaag aagagatagt 1140ttccggtgat ttctccattg atgttcccca
gtccaacttc acccatcatg ttggtacctt 1200tgcaagccag aagcttgcac agacagctaa
tgggactgca ttcgcggctc acgctctatg 1260gcatttcgcg caaacctggt ttttcgagtt
cccacactac aagccaaacg atgatcgtgt 1320cagtctctgg gctgaaagtt acggaggcca
ttatggtcca ggcatctttc ggttcttcca 1380acagcagaat gacaaaatcg cagaggggac
tgcagaagac ggtgcacagt atttgcatct 1440cgacacgctt ggcattgtga acggcttgat
ggatatggtg atccaagaag aggcttacat 1500tacttggcca tacaataacg taaggctcgc
cccttcttca ttcaactcgc gtaatgccta 1560attcagttca gacctacggc ctcgaaatct
tcgataaacc cctctacgaa gaactgatgt 1620ataactggac gcatccagga ggctttcgcg
atcaggccct cgcctgcgaa gcggctttga 1680aagaacgcga ttccggcttg cctcactcag
ggaagaatat ctctgaaatt tgcggaggcc 1740ttgcactaga atggggagat ggccccatca
cctactacca caccttcaat cgcgggtggt 1800acgacatcgc ccatcctaag aacgacccat
tccctgccaa gcacatgctc ggatatttga 1860cgcaggagtc cgtccttgcc gctcttgggg
taccagtcaa tttcacatcg tcttcgagtg 1920ccgtggctac acagttcata aaaacctttg
atatcgtcca cggcggcttc ctggatgcaa 1980ttggctacct cctcgacagt ggtgtaaaag
tacacatgat gtacggagat cgtgattacg 2040cctgcaattg ggtcgggggc gaaaaagcca
gccttgcagt tccgtattcc cgtatcaccg 2100aatttgccga cacgggatac tccccactcc
ttacgcccga cgggatcagc ggcatgaccc 2160gccagctggg caactacagc ttcactcgcg
tcttccaagc cgggcatgag gtcccctcct 2220accagcctgt cgcggcgtat gagatcttca
tgcgggcgac attcaacaaa gatatcccta 2280ctggcctctt ggctgttgat gacgaattcc
agtcggttgg acctaaggat acgtggcata 2340tcaagaatat ccctcctatt atgccaaagc
cgcagtgcta tgttctaagt cccggcacgt 2400gtaccccgga ggtttgggag accgttttga
acggatccgc gacggtaaag gattggtatg 2460tcgtggatga tagcgcgggt gttgaggacc
acgaggggtt cagcattctt ggaggggatg 2520agttgtagta gcttagaagt ctaccggaaa
gccttgctag ctcttaaaat acgacacaaa 2580tagttgagcg acatagtcta ctctctcact
tacgaacacg tgatgaatct ctccagcaat 2640aaagtggacc aatcaattgg aggaaaatat
atccacgatg gtcggacggg ccaacgaaaa 2700gggccaacta gcgcccctct gaggatgatg
gtctagtctg caggatatga gattgttcac 2760atcttatggg tcttcaactg cacaaattag
gcggaggtcg cctcaccgaa agagacgtaa 2820gctgcttttt ggtggattca ttattagccg
ttatgccatt tactaaaaca ccgcagttcc 2880agatcgcgta gatggcctca aagtgttcgc
cagatttgat tgagggctac ttcgcccaag 2940cctatggcct gtagatgcca atttcttcaa
tacaaagata ggcgaaacat gttgaaaatc 3000catcatattc aattaaattt caaaacatag
cgagacatta agtattacat atatttcaat 3060gaatagagac ttcttgcaca
3080482520DNAAspergillus niger
48tctcgcctgc ctcgaccttc ctgctatggc aattctttcc ccaagctgag cggttccatc
60cgcttcccga ccgccaaact tatcaacccg agcgatacgg agaggggaaa ggctatcacg
120gcaatgattg cgcaactctg cgataagata accccatgtt ttgacgtagc ctcggccctg
180tcttctcctc attattatcc tccgattccc ccagggttcc tgtcttctgc agaaagtgct
240ctcggaatcg aagatgacca ggtttcaatt gcttcccctt gtcgcagggc tgcttgcccc
300ttcaattgca gcccttagca tcccttcccc gcagcagatc ctcgattctc tcactttcgg
360agagcacacc gacggctttt gtccgctggc acccaaggtt gaggttcctg acgatggttt
420ctttccagct ctcaagttcg tagaagatgc ctcgttcaag tcgcgccaag tcaatcgtct
480ctccagggcg gttcaagttc cgaccgcaat cgacgactac atgaaggatc cctacgacga
540aaagttcgcc ccattcctcg acttccagaa gctcctgcag accctctttc ccctcacgta
600cgtccccagt ccccgtcttc atggctccat ctaacgcaat ccagccactc ctacgcccgc
660gtagatcaca tcaaccgatt tggtctcgtc ttcaccctca atggcacaga tgactcgctc
720aagcccctgc tattcaccgc gcaccaggac gtcgtgccca tcaacgaccc tgccgactgg
780acctatcccc ccttcgatgg ccactacgac ggcgaatggc tctggggccg cggtgccagc
840gactgcaaga acgtcctgat cggtctcatg tccgttgttg aagacctact ctcccaaaag
900tgggagccaa cccgcacagt cgtcctggcc ttcggattcg acgaagaatc ccacggcttc
960ctcggcgccg gatccatcgc caaattcctt gagaagaaat acggaccgga cagcttcgaa
1020tttatcctcg acgaaggcgg catgggcctc gaagttctag acgacaacaa caacggcgtc
1080gtctacgctc tccccggcgt tggcgaaaag ggcagcatcg acgttgtgct cactctggcc
1140gtaccaggcg gccacagctc cgtgccccct ccacacacgg gaatcggcat catcgccgag
1200atcatctatg agctagaacg ccaggacctc ttcgtccccg tcctagacac tcaccacccg
1260acccgcaaga tgctcgaatg ccaagtccgc cactccccct cgcaagtcga accgtggctc
1320gcctccgccc tccaatcaag cgactacatc tccctagcag agaaactggc ctcctcgcgc
1380ggcgacaagt tccgcttcat cctccaaacc tcccaagcag cggacatcat caacggcggc
1440gtcaaatcca acgctctccc cgagaaaatc aacgccctcg tcaactaccg catcgctctg
1500caccaaaccc cagacgatat caagaaccgc gctgtggaga tcatctctcc catcgtcaag
1560aaatataacc tctccctcac ggccttcccg gaaagcgaca ccgttgaccc ctccctcaac
1620aaccacctca cccttactac cctcagcggc gccctcagtc ccgccccggt cagcccaacg
1680gacatcgaca ccgacgccgt ctgggcccgt ttctcgggcg tcactcgctc ggtcttcgaa
1740tctgtcccta gtctcgaggg cagaaaggtc gtcgtgagcg gcgacatcat gaccgggaat
1800acggatacga gattctactg ggctttgtcg aggaatattt acaggtggag tccgtcgagg
1860gcgggtaaag cgctgaatat tcatactgtt gatgagagga tcgatattga tattcatctt
1920gaggcgatga tgctgtatta cggtatgcat ctctcctttt atggatactc ctatcagact
1980aacttgtatc tctagatctt attcgctctt tcgatggacg gaccgattca tctgtcattt
2040ctgctgcgtc ggcagctgct gatgatgaac ttgctcacga cgtgctgtga gtctgtagga
2100actctcctac tctttagggt ggctcaggat gaatgatggt tatgaatcat gtggtctact
2160gcgccaaaag ttcatgtctt tcctggagtt caacgagaac cagatctata tttccacgaa
2220tagctccagt atatagccaa atgcaaccac gtatttgtct atcgatctga cagtctaatt
2280ctttacctta tactaggggt aagaaagggt aagaaaataa aataatattt gcagcggcct
2340atgaagataa ttatgtactc gacgttgccc aatgcactgt agcccctctc cctgtaacag
2400gagtagccat aaaagcaaaa cgaaagtggt aagaagacgg gggcaatatc cctgctttag
2460caatatacaa caatagtaac aatcgagagc tcaatatgaa tgtataacga gtgacctgca
2520492730DNAAspergillus niger 49tttagttgtg atctcaatcg caaatggaca
ggagatgata ttcatgtcat aatcagagtg 60ccatgtcagc gaccatgtct tgccaagagt
catcatttcc ggtaaaaccg acggaggtaa 120aaccccggtc attcactagt tcgagggagg
gaatagtctc gcaggtctct tcttctttct 180tcctcggggt ggcggatgga tcacttcttc
atttgatggt tgcctgccaa cgacccactt 240gactaggttt cccccttgat gggattcgtt
ggttttacca gacagagcct cttcacagcc 300cagcgcgtgt tccgcaaaag caacagccca
agggtctgaa ggctgacatt ggagcgggtc 360tgacgcagga tggcttgctg ttaaaaaaac
tcaggaacag gccaagacga agcgaccgcg 420caaggctgag cagttctccg atttccccgt
gcccctcctg cgcctcccat gggcgacttt 480tgatcccaga tccatcgttt ctgcaccgac
catataagaa gctccgcacc cccaaactcc 540cctggcaagt tcctgccccc aaattacgtg
agacagggaa ccatcacagc aacaccatga 600agagcaccac tcttctttcc ttggcctggg
ctgcccagtc cgcctattcc ctctctatcc 660acgagcgcga tgaacccgct actcttcagt
tcaactttga acgtcgtcag atcgccgacc 720ggtcccgtcg gaagcgatcg acggcctcgg
ccgacctcgt taacctggta tgttcccatc 780ccgagtctca aatcagggga attatgacgg
tcgctaatgc aggtttctag gctacgaatc 840ttggctacac gatgaacctc acactcggca
ctcccggcca ggaagtcagt gtgacgttgg 900acaccggcag cagcgatctc tgggtcaatg
gggccaactc gtccgtctgc ccctgtaccg 960attacggctc ttacaactca agcgcttctt
ccacctacac cttcgtgaac gatgagtttt 1020atatccagta tgtcgacggc agtgaagcca
caggcgacta tgtcaacgat actctaaagt 1080tctccaatgt gactttgacg aactttcaat
ttgccgtcgc atatgacggc gactccgagg 1140gtaagtcttc gctattccct cactttcatt
ttacactttg ctaacggttt taccatgcag 1200agggggtcct cggtatcgga tacgccagca
atgaagccag ccaggccacc gtcggtggtg 1260gtgaatacac caacttcccc gaagccctcg
tcgatcaagg cgcgatcaac tggccggcct 1320acagtctatg gctcgatgac ctcgacgaag
gaaaaggcac cattctgttc ggcggagtca 1380acaccgccaa gtactacggc agcctgcaga
ccctgcctat cgtctccatc gaagacatgt 1440acgtcgagtt cgcggtcaac ctgacggccg
tgcaccttga gaagaacggc aactccgtct 1500cggtcaacaa cagcgccacg caattcccca
tccccgccgt gctggacagc ggcacggccc 1560tgacctacat cccgacctcc gccgcagcca
gcatctacga ggccgtcggt gcccaatacc 1620tgagcgagta cgggtacgga gtgatcgagt
gcgacgtcaa ggacgaagac ttcaccttcc 1680tgttcgactt tggatccttc aacatgagcg
ttgacatcag cgagatgatc ctcgaggcca 1740gttccgacat gaccgacatg aacgtttgta
cgtttggcct cgcagtgatc gaaaatgagg 1800ccctgctggg cgataccttc ctgcgcagcg
catacgtcgt ctacgatctc ggaaacaacg 1860agatctccct ggccaaggcc aacttcaacc
ccggcgagga ccacgtcctg gagatcggca 1920ccggatcgga tgccgtgccc aaggcgacgg
gggcgacggc gaccggcgcg gcagccacat 1980ccacggcctc gagcgacaag tcggacaagg
agagttcggc tacagtgccg cgcagccaga 2040ttgtctcgct ggtggcggga gtcttggtcg
gtgttttctt ggttctgtaa atatagagat 2100ttcacgttgc atgttgatga tacataccat
agatttgctt ctaattgccc cttacatctc 2160gaaatgtttt ctgtttttag cttgatatcc
catcgcttca ggtttcagat atggaaatga 2220aatgaaaatc ggaatgcaag atctgatcga
atagttgttt cactgccgcg tggctggtcc 2280gatcggcacg aagccagtaa tcgacaacca
atcatacatt gtgatttcct gtcaacctcc 2340ttgcgactac tactaatttt atcatatcat
gccgttgtga cctatcctac tcggtgtccc 2400cacaccgatg atagccaccc gcgaccggat
aatcggtaac ctgccaagag tcgaacggct 2460ttcctgtgga aagggtggct gcaacctgtt
tgtttcagtt tttgtgttgc agaattgact 2520atccttctga cctgtgaatg atactgggat
ggtgacgtag tgaaaatgtt ggtggagcga 2580ttaaggtttg tgcctgtcag cctcgtcaag
tccatcatca tgtcttttcc ggactatcac 2640ttcctctact ctgaagtacc gggactcccc
gccagtattc gtttcgccaa tcagagcatt 2700cgatgtgaac cggattgaag agtgcccggt
2730503231DNAAspergillus niger
50ccgacgcgac ggcattagtt cctcgatagc gtgggaagac catggagaga ccggccgata
60agctgcagag gagctttctg tgggctgacc agtgcctatc agcaaatgga cacgcggtga
120cacccccgca atcgcaagac cgcctcgaca ccgtccatcc aggcaccatc gcaacggcaa
180accagcctct cctccaatca aaccatcgat gtgatattgc tagcggcggt gagaacagca
240ggtcgaatgt cgcaattaat tctcgcatcg tccctcactg caaggggccc actaccagag
300ctctgcggcg ttgttattta aatccgttgt tgggaccttt cggtcatcag ccactgtgtc
360tggtcactat cgcgctggtt caacctcaac atgttggtcc gtcagcttgc cctggctctg
420gccattgcgg ccttgtccga tgccattccg acatccatca agcatgtcct gcacgagaaa
480cgtcacaagc ccgcatccga ctgggtgaag ggtgcgcgcg ttgagagcga tgcggtcctg
540cctatgcgca ttggccttgc ccagaacaac ttggacaagg gctatgactt cctgatggaa
600gtgtgagtca aaatctacct tttttatgct atatgcctag ttttaggatc gcaatggatg
660acctcgaaac atgctgacat gagattggcg cagatcggac cccaagtctt ccaaatacgg
720ccagtactgg tcggcagacg aggtgcacga catcttttcg ccatccgagg aggctgttga
780ggcagtgaga gaatggcttg tcgcctctgg tatccatccg tcgcgggtgg tgcactccga
840caacaagggc tggctcgcgt tcgacgccta cgcccatgaa gccgagaggc tgttcatgac
900ggaattccac gagcacgaga gcgaccgaag tgctaagatc agggttggat gcgaccagta
960agaagattct tctatcacct tccatgagta gctattaatc ggaatctaga taccacgtcc
1020ccgaacacat ccagaagcac atcgactaca ttacccctgg agtgaagctc acccaggtcg
1080tgaagaggac caacaaagtc aagcgtgctt cccaactagc tcactcttcc aaggccaagt
1140ctgctgccca aggtccgcag ccactcccca acaaggccaa gttcctgcct gaagacctcc
1200gcggctgcgg ttacaacatc accccctcgt gtatcaaggc cttgtatcag atcccagacg
1260ctaagacggc gaccccgaac aacagcctgg gtctgtacga gcagggtgac tactttgcca
1320agtccgacct cgacctcttc tataaggagt atgcgccgtg ggttccccag ggtacctatc
1380ccatcccagc cctgattgat ggcgccaatt actcggttcc ttcctacagc tccctgaaca
1440cgggtgaatc cgacattgac attgacatgg cgtgagtcat ttctgcacct tgtcatcaga
1500cccctactga cgttttgaag ctactccctg ctctaccctc agcaggtgac cctctaccag
1560gttgacgacc agctctacga accagtcgag gtcgacacaa ccaatctgtt caacaccttc
1620ctcgacgctc tcgatggcgt gagtacagac ctcgttctca gtcttaccca gctaacaccc
1680ctagtcctac tgcacctaca gcgcctacgg cgagaccggc gatgacccgt cgatcgaccc
1740cgtatacccc gacacccgcc ccggcggcta caaaggtacc tacctacacc acctcttccc
1800catacaatcc aacctaacac accaacagga aagctccagt gcggcgtcta taagcccact
1860aacgtaatca gcgcctccta cggccaatcc gaagccgacc tccccgtcag ctacaccaag
1920cgccaatgca atgagttcat gaagctcggt ctacagggac actccatcct cttcgcgtct
1980ggcgactacg gcgtcgcgtc tttcgccggc gacggtgacg agaacggctg tctcggccca
2040gagggcaaga tcttcaaccc ccagtacccc tccaactgcc cctacgtcac ctccgttgga
2100ggtaccatgc tgtacggcta ccagaccgtc aacgacagcg agagcgtcat gcacgttaac
2160cttggcggaa ccgcaagtaa cttcagcact tctggtggct tctcgaatta cttcccccaa
2220ccggcatatc agtttgctgc tgtggagcaa tacttccagt ctgcgaacct gtcgtatccg
2280tattactcgg agtttgaggt cgatgttaac acgaccaagg gtctctacaa taggcttggt
2340cgtgcttatc cggatgtctc ggcgaatgga gcgcatttcc gcgcttatat ggatggatac
2400gattatcatt ggtatggatc gagtttggcg tcgcctttgt tcgcgtcggt tcttactttg
2460gtgagctttg tcacccccca ttactaatta ttgacacatg gctgaccgac ttagctcaac
2520gaggaacgct tcgctatcgg caagggcccc gtgggattcg tgaatcccgt gctttatgct
2580tatccgcaag tgctgaacga tatcactaat ggtactaatg ctgggtgtgg aacttatggg
2640tttagtgcta ttgaggggta agtgctcagt acttggttct gtcaggaggg gtgtgctaat
2700tgatgactat agatgggatc ccgctagtgg tttgggtacg cctaactacc cattgatgaa
2760ggagctgttc ctctctttgc cttaggattg aacggtgctg tgtcagaggg tgataggtgg
2820tcaagctgtg tatatatgtc tgatggggaa atatttacga tcataggata atgtgtcgac
2880gagcatgaat ggccaattat ctcgcctgtc accgtgaata aggtcaaatg tagatcggtt
2940taatagttca actacagaga attcttggat attgtcaaat gttgactatt cgctgtctct
3000ttatcgtcta atgtataata tcatcaaata acctaaccca agggatatca aaaacataag
3060aaaataataa accggtacta tgtgtcgaaa aaaggaatgt ttgtgaattt tttaaaaccg
3120ttcatacctc ccgtccatgt ccaccaaatg cacagcgctg gtcgaatccc tctccgaccg
3180tgtcgtatag tcaacggaga tattcactga cttggaaatc ttgctgttgg t
3231512660DNAAspergillus niger 51tcctgagcaa gcagctaccg gtaatctgag
acctaatcct ggtaagtgga tcgagttcat 60ttacctcatc atctaacctc gcttcatgat
tcgcaggcgc tttccgcgac tagtcgatgc 120cagtatccat ttgcctgcag atccgatgct
tccaccgacc accgttactt ctctccagac 180ccccatccgg ctttccgacg gcctcctctg
catcccctca caaagcaagc ggagacctgc 240ctgaataggg agagtttttt caatgagccc
acacgcagtt acctcgcttc cgcatgagat 300gggtccattc ttgacaatgc tgcctaatgc
gcagaagggc cgtcatgcgc ctggagaact 360acataaatag ggaccacgca tgccccgacg
atgtatgaat tgttcaaact tctccgccca 420agtgagttct cttttcattc cattccgaag
ggagaatcac cagtatgcgg gttaccacgg 480caattgcttc attactactg gtcggctcgg
ccaccagtct ccaaaatcct catcgtcggg 540ctgttccgcc ccctctctcg catcgcagcg
tagcgtctcg ctccgtgccc gttgagcgcc 600gaaccaccga ctttgagtat ttgactaaca
agactgcaag tgcgtgattc cgttttttaa 660ctaccgcatt tatcgttcta agatcaattg
caggattcct ggtcaatggc acaagcatcc 720ccgaagtcga tttcgacgtc ggcgagtcct
acgccggcct tctccccaat acgcccactg 780gcaattctag cctattcttc tggttcttcc
cctcgcaaaa tccagaggcc agcgatgagg 840ttagtggtcg ctctgttttt tccggtcatg
cgtcagccag ctaacaatta acaaagatca 900ccatctggct caacggcggc cccggatgta
gctccctaga cggcctgctt caagagaacg 960gcccattcct ctggcagcct ggcacttaca
agcccgttcc taatccatac tcatggacca 1020acctcaccaa tgtggtttac atcgaccaac
ccgccggcac aggcttctcc ccgggcccct 1080cgaccgtaaa taacgaggaa gacgtggctg
cccagttcaa cagctggttc aagcacttcg 1140tcgacacctt cgacctgcac ggccgcaagg
tctacatcac cggtgaaagc tacgcgggca 1200tgtacgtccc ctacattgcc gatgccatgc
tgaacgagga ggatacaacc tacttcaact 1260tgaagggtat ccagatcaac gacccgtcca
tcaacagcga ctcggtcatg atgtactgta 1320tgtttccctt catatacctc cacctccacc
accaccacca ctaacaacat cacccaccag 1380cccccgccgt ccgccatctg aaccactaca
acaacatctt ccagctaaac tccactttcc 1440tctcctacat caacgccaaa gccgacaagt
gcggctacaa cgccttcctc gacaaagcca 1500tcacctaccc accccccagt cccttcccca
ccgcccctga aatcaccgaa gactgccaag 1560tctgggacga agtcgtcatg gccgcctacg
acatcaaccc ctgcttcaat tactaccacc 1620tgatcgactt ctgcccctac ctctgggacg
tgctcggctt cccctccctc gcctccggcc 1680caaacaacta cttcaaccgc tccgacgtcc
agaagatcct gcacgtccct ccaacggact 1740actccgtgtg ctcggagacc gtcatcttcg
cgaacggcga cggcagcgac cccagctcct 1800ggggtcccct acccagcgtc atcgaacgca
ctaacaacac tatcatcggc cacggctggc 1860tcgattacct cctcttcttg aacggctcgc
tcgccacaat ccagaacatg acctggaacg 1920gtaagcaagg gttccagcgt cctcccgtgg
aaccgctctt cgtcccttac cattatggtc 1980tggctgagct gtactggggc gatgagcctg
acccgtataa ccttgatgct ggcgctggat 2040acctgggtac agcgcatacc gagcgcgggt
tgactttcag ctcggtgtat ttgtctggtc 2100atggtaagtt tattatatcc ccttggaagc
ggtatgatga acgttagaga gtgctgactg 2160ttgcttcctc ctcgtgatag aaatcccgca
gtatgttcct ggtgcggctt accgccagtt 2220ggagttcctg ctcggtagga ttagtagtct
ttcggcgaag gggaactata cctcttgatt 2280tcagcggatg acgacaaaaa gatgatgagg
aagatgattg gatgctttca gatgggaatg 2340cgtgcatagg agatgagatg agatgatgta
tcattgaggt gtgatgactt gtacatatgt 2400agatcggtag taaaagggat actataggat
atggtatttg atgtatcatt tatgtacacc 2460acgtggattc aattgagggg agcttcaatt
cctggttatt acttatcata tttccaacat 2520gtccgcgtat aaccggtaac aactaacggc
ttcatgtttg tcaagtgact gtcttggtac 2580aatactactg tttctatatt ctactgttgg
taacttaatc tggacatatt ctatatccac 2640gtacagattc tcgctagatt
2660523150DNAAspergillus niger
52ccaatcaacc acggatgatt gggttcaggt cctggaggca cggagtcgga ccaagacact
60aaggtacagc actagcgaga caatgtgtta tcgctattat tggcaaaatg gccgcgggat
120ctcttatgca gggttcggct ccatcctccc ccctcttcct atccagtcaa tccgcctcgg
180ttattgaagg agatcctgag ctgtttaact gacgcctcac cgatcaggcc ggaaatggtg
240gcgggataca acatcgtttc cacacaatag tgcttgtctc ctgcgatctg catggcatgc
300taatctccgc cagcatgtat cttctatcca ctggatatga attttcctcc cctcacacca
360tgtgggcctg ggggttttcc ctcaaacttt gtcgctcatg taacgatgta tataaagccc
420tgaggatggc atcccccacc catcggtctt ttgctgaccg ttctccttga agaaattctc
480gagtggcttg tggtgcatgt atagatttaa tcttcgaggg ttattaacta ggtatagctg
540tgactaagtc tgtccttgca ttgaacaaca caccatgcgt ggctctcggt tggtgctctt
600gttgcccctg gctgcactta gttgtgctat gcccgagaat gaatggtcat ctacgataag
660aaggcagtta ccaaaagcgt ccactggcgt caaatcgata aaaaccccaa acaatgtcac
720tatcaggtat aaagaaccag gaaccgaagg aatttgtgag acaacacctg gggtcaaatc
780atactccgga tatgtcgatc tttcgccaga gtcgcatact ttcttttggt ttttcgagtc
840acgccgtgac cccgaaaatg atccagtgac tctgtggctg aatggtggcc ctggaagcga
900ttccttgatt gggctttttg aaggttggcc aaatatcctg acggaaaaga taaaattcag
960cttgcatgtt ctgacgcctt cacaacagag ttgggtccgt gtcacatcac accagagtac
1020gaatcaatca tcaatcagta ctcctggaac gaggtcacca atcttctttt cttgtctcag
1080cccctcggtg tgggtatgga atattgctgc cttcatacat cctgagtaca ttgcttacgg
1140tcttatctgc gaagggttct cttacagtga aaccgaggcc gggtccttga atccatttac
1200tggagccgtc gagaacgcct cctttgctgg agttcagggt cgatacccag ttattgatgc
1260cactatcatc ggtaagttgt ccggtttgac tctcacctag cattctcctc aatgtcctac
1320tttacagaca cgaccgatat cgctgcacgc gcaacctggg aggtgcttca gggcttcctc
1380agtggcctgt cgcagctaga ttccgaagtc aagtccaagg agttcaacct gtggacagag
1440agttacggag ggtgagtgca actttcatac cagaccgacg taagctgact tgatcaagac
1500actatggacc agcggtaggt tgtcttttct ggttgcacac atattgatct aatgaccgaa
1560gttcttcaat catttctacg agcaaaattc gaagatcgct agcggggaag tcaatggcgt
1620ccaactgaat tttaactccc tcgggattat caacggcatc attgatgccg cgattcaggt
1680acttagaaat gcagctcgcg cagaggctgc ggcctagaag gacatcgcta aagtaattaa
1740taggcagact actacgcaga ctttgccgtt aataatacat atggaatcaa agctgtaagt
1800ttaaatacac gtacatcgtg gatttaagat caaccgtgct catgcttgct aggtcaatga
1860cacagtgtac aactatatga agttcgccaa cacgatgcca aatggatgcc aggatcaggt
1920tgcttcgtgt aaattgacca ataggacctc gctttctgat tatgctatat gtacagaagc
1980agccaatatg tgcagggaca atgtcggtga gtggttctac tgtttctctg caggggtgca
2040atgatgaagg actttgctaa gctgtcatgt acagaagggc cttactacca gtttggcggc
2100cgtggcgtgt atgatattcg gcacccctac aatgtaagtg gcaaggataa ggattgtact
2160ttccgaacag ggacactgct catatgtcaa cgtaggaccc gaccccgccg tcctactttg
2220ttgactacct caagaaagac tcagtcatgg atgctatcgg cgtggacatt aactacaccg
2280agtccagcgg cgaagtatat tatgcattcc agcagaccgg cgactttgta tggccgaatt
2340tcattgagga cctcgaagag atcctccaac tccccgtacg cgtgtcgttg atctacggcg
2400atgccgacta tatctgtaac tggttcggcg gtcaggccat ctcactcgca gttaactacc
2460cccatgcagc tcagttccgt gcagcgggat acacacccat gacagtagat ggggtcgaat
2520acggtgagac tcgcgagtat ggcaactttt cgttcacccg cgtatatcag gctgggcacg
2580aggttccata ctatcaaccg atcgcagcgt tgcagctgtt caaccgtact ttatttggat
2640gggatattgc agcgggtaca actcagattt ggcccgaata tagcaccaac gggacatcgc
2700aggctacaca cacggagtcg ttcgtgccac tgtccacggc gtcgagtacc accgtcaatt
2760aggattgggg gaaatttttc cctctttggt atggctaatg ctgtgttatc cattccgata
2820cttgtccata acctaaaaga gtggcacgga agctttccta gacatgcttg ctctagtcaa
2880cttttatcct accactgtgt ggtcctacca taatgtcatc ctaaacttat caggtgtctt
2940acatcatttt gcagtgacca taaaagtcat gtcatattaa gtccattacg gtagttcgta
3000acttgctgat ggcttacgta attgtgcctc agcaggatgt cgtgatacgc tccaaaaccc
3060aatctgatca tggatatcca gcggaataac acaaaagaaa aaataaacat aaccaaccac
3120caaaatgaca gaggctaatg cctggacaac
3150533221DNAAspergillus niger 53ccatcagctc ttgttgttat gttttatgtc
atagtgtaat gctagcctta gcgtgcgtgg 60tgtttgaatt tcagattggg catggatacc
tgggcgtctc gggatggata ctgggacgtg 120gtaagatgct tggaagcgtt gattttgagt
ttgacaatat atccctatgt ttctcttctc 180cacacccttt taccgtaatg cgacagcatt
gtgtagagtg cattgcctta attagaacct 240acgaatcctt gatttgtata tgacacccga
tcaccatcga gccacctaca tcatggcgcg 300ctctcagatc ctccgaacca aaacagtaat
gtcgatcgtc gcgcctgaac caaggctgat 360tttccctgct gaagcgtgtc ataaagtgaa
acatactttg tagataaact cggcaattaa 420ctgccactta ccccatacat cgctaccgga
gagtggagcg agtgcacggc catcaagtga 480aactgataac cggcgatccg ccggcttatc
ggcgatggcg ttatcagagg gaccttagta 540ggttgcacag ttcactagta tttattacta
agtacttcct tcgccttgct tattgcccat 600gattcgtcct tctctttttc tttggttaat
tgaactactc tccatgagag ttctgtacag 660gttgagatag accaaccacc accaccatgc
cttttccctt ttcgtccgct cttctcggct 720atatcttaac tacgagcact actctcacct
ccctagtcgc aggacagtat taccctccga 780cgcctgagga tctcaccgtt attcattcgg
agatattccc tggtgcgagg atctcctata 840agcaagtgag aaatacacca cccttctcct
caatcccaat ctaacacccc atactaatac 900tatgtgacca gcccctcggc atctgcacca
ccaccccctc cacccccagc tactccggct 960acatccacct ccccccacac acccttacca
atctctccat tccaggaatc agcatctcgc 1020aaccataccc tatcaatacc tttttctggt
actttccttc ccgccatcac cacaacaatg 1080atacatcccc actcaccatc tggatgaacg
gcgggcccgg cggatcctcc atgattgggc 1140tatttcaaga gaacgggcca tgtactgtga
atacggactc gaattccacg gcctataatc 1200cctggtcgtg gaatgagtac gtcgatatgt
tgtatattga gcagccggtg cagacgggat 1260ttagttatga tgtgttgagg aatgggacgt
tagatttggt tagtggggag atagatgtta 1320gtattagtga tggtgagagg gatggagtag
gacagaatga gacgtttttg gtggggacgt 1380tgccgagtca ggatgtgcat gggacggtga
atgggacggt taatggggga agggcgcttt 1440gggttgcgtt gcaggtttgg ttgggtgaat
tctctgaata tgtttcttct gttgacggga 1500atggtggtgg tgatgacagg gtgagtatat
ggacggagtc atatggggga cggtatggac 1560cggcatacac ggcgctcttt caggagatga
atgagaggat tgagagtggg gaggtaagca 1620ccgggaagaa gatccatttg gatacgctgg
gcattatcaa tgggtgtgtg gatttactcg 1680tgcaggtccc ttcgttccct gagcaggcgt
ataacaatac gtatgggatc gagggaatca 1740atcgcacgct ctacgaccgg gctatggata
gttggagcaa gcctggcggg tgcagggata 1800tgatcatcga gtgtcgcgat gctggcgagc
tcggagatcc cctcatgtat ggcgacaatg 1860agacggtaaa tagcatctgc gaggaggcgt
cggactactg ttcgcgggag atcaagagcc 1920tgtatacgaa tacctccggg cgaggatact
acgacatagc gcatttcacg ccggatgcag 1980ctctcgtgcc ttacttcgtc gggttcttga
atcgcccatg ggtgcaaaag gcacttgggg 2040tcccggtgaa ctataccatg tcgtcagagg
cagtggggaa cagtttcgcc tcgacgggcg 2100attatccgcg aaatgatccc cgcggaatga
tcggggatat tggatacttg cttgactccg 2160gtgtcaaggt ggctatggta tatggggacc
gggactatgc ttgtccgtgg cgcggcgggg 2220aagatgtcag cctgctggtg gagtacgagg
atgcggagaa gttccgtgct gctgggtatg 2280ccgaagtgca gacgaagtca tcctacgttg
ggggtctagt aaggcagtat gggaacttct 2340cgttcacgcg tgtctttcag gcgggccatg
aggtgccatt ttatcagccc gaaacggcgt 2400atgagatttt taatcgcgct cagtttaatt
gggatattgc gacgggaggc atttctctgg 2460agcagaatca gagctatggg acggagggac
cgtcgtcaac gtggcatatc aaaaacgaag 2520tgccggagag ccctgagccg acgtgctatt
tgttggcgat ggattcgact tgtacggatg 2580agcagaggga acgggtgctg agtggggatg
cggtggtgag ggattgggtt gttgttgatg 2640atattgaggc tgaaagctcg ttcagcggtg
ttggtgatca gctggcacag gtccctttgg 2700gacattgacg ttggttgcca tatgttgaca
gtgttggtgg atgaaagtga tatagatgga 2760taaagttagt gcataagtgc atagctgact
aagaatgagc atcattcata tatggataag 2820tctggctagc tcgatccggc aagtggccgg
ctgactgcct aaagtggaag actggaagcc 2880ttcagatttc catcagtcca tctcaatatc
tccatctggc ttgtcaaact cctatatcca 2940ccggccggac ccttctatta ttaacagcgc
cgctatcacc cggacaatcc gtatggcatc 3000acgggttgac ccctccattg cggccatcga
accttctcga accttctatt ggtcgaccgc 3060tgcggtcaac gtcacacgtc accgcaggct
cacctgaaag ccgccggcgt tgcactatac 3120atcagccctc gtcctcacac ctggctttct
ccgctaatcc tccaagtaca cccttcactg 3180tgccttgatg tcttgaatca acatctataa
aaactgcatt a 3221542590DNAAspergillus niger
54gagtgagtaa attgatctaa cggcgtcgcg ctgacaaaca tcatagctat caatcagaaa
60aagcgaagca tatttcacat acgatcggat agcctaccaa ggaactgagg atatacgtac
120agctttcgct tggctgacac gtaaagtcat ttggtgattt tggagcgact ttggatctcc
180ttccatgtaa gttataacct accgcaattc caaagtatag ctagtagctg tcagtcactc
240aagaacttgc agcaaggcga tggcaagaca aaacaatcga tggtacggag tagatagaag
300acactaaagc actttcacat gacactgttg acatagtgct tatgctcttt acactcaaca
360tgagtcactt tgataaggtg atttcgccag atggtggcat tattgactcc tgattggatc
420caccgataag gaaccacttt ttggctggca atgtactatg ttattgacag gcatcgaact
480tccccaaatc tcaggctggg ccgccagcaa atggcgcggt tccggactcc ggcgggggct
540gacagggcca accgttggct gcttgtccct tgccgccgga gagcttattt aactcttgtc
600tctcctcttc ccccacaaag ctcactcagc actcaaggcc tcaccgcgcc atagtcccgc
660tcttgatcat ccttagcaaa acggctgtaa atcatgagaa catctactct tttgctcctc
720tggagcactg caggagcagc tttggcttct ccgtacccgc ttcccgactc gcaagtagtc
780ttcgccgcgg atcacgaggt cccgaataca cagggcaaac acgtcgtgga cgaggccata
840ctctcggcgc tgaacgctca ttctgaccca gtcgctgcaa tggtgtctct acgtcccgaa
900actgcagctt ttctagctga acctcgtctc ttgcacattc ggggcgaaga gaaggcggaa
960tggatgaccg aaggcgacaa gctgcgcctc cgccaacgcg gaaagaagtt catggacatt
1020accgagcatc aggacttcta cgcagagcag gcgatggctt cgtttgctgg ggatcctagt
1080taatctccct tttgtcgagg taattaatac tgtgttaacg ccctttagat cttcccaagc
1140tgtcccataa aggtctcgtc aagccgctgt tctctcaaat cgagacggaa cgaatgcacg
1200atatcctgca gcacatgacc tcctactaca atcgatacta cggtgattat cacggcgaga
1260tgagctccga atggctgcac gactacattg ctgcggtatc tccaccccct acagccacaa
1320tttttgaaca tgaacttact caaccaagat catctccaaa tcgcctttcc gcacccacat
1380ctctctcgaa tacttcaccc atcctttccg ccaatcttca attattgcac gcttcgagcc
1440taaagttcgc agcttctccc aacctttgac catcattggt gcgcaccaag attcggccaa
1500ttatcttttt cccctgctgc ccgcccctgg cgctgacgat gactgttccg gcactgtcag
1560tatcctcgag gccttccgcg ttctggcgga gaatggctac acgcccaagg acgggcctgt
1620tgaattccat tggtatgcgg ctgaagaggc cgggctactg ggcagccaag ccatcgcgcg
1680gtacaagaag gagcagggcg ctaaaattga tgccatgatg gagtttgtag gttcccatca
1740tccaacggac ttggaatcac ttttgacttt cggttattaa catctgcata ggatatgacg
1800gcttttattg cccgtaacgc caccgagacc atcgggtttg ttgcaaccca agccgatgca
1860gcgctcacaa actgggccct caacctcagt cgagaataca tctccattcc ggcggaagtc
1920tatgaacttg gcccgtaaga ccccagaata tccccagagg catgtcaagt gctgaatgat
1980ttcttagcaa cgctggatcc gactacatgt catacactaa gctcaactac cccgctgcct
2040ttgcatccga aggcaacccg ctcgctgggg gctctttccc gggtgaaatg gacccctacg
2100tacacggcat caaggatagg atggacgttg acgatgaaac gggcgtcttc tctatcgaag
2160taagttgacc gcactcaacc tgtccatctt cttgctaacg atttgccagc acatggctcg
2220gttctccgag ttggctatcg catttgttgt cgagcaggct gggtgggata atacatggcg
2280gtagagtata tacagggatg tttccagctt catagcatat agagctggta tatagtcatt
2340gtgcttacaa tatataccat tctattgagt tcttgacaga gttattatgt cagaatggta
2400atcagtctaa atcaatgcgc ccttaaatca agaagtacac tagtgcccat gaccaaaaca
2460atttccactt acacatacca cctatctgcc tcactacgaa acccttacca tacaatgttt
2520acgttgtgct cctttcaata ctccatttcc atagttcatg ctatcgtggg ataaacatgc
2580acatatcctc
2590553290DNAAspergillus niger 55gagaggcaga aggagtcatt tatcacttgt
attccaatgt attttccatt tatagatact 60gcattcaaat gcaccgttta gcatagcatc
ccacattcta tttcattcca atctcatgcc 120attgccatcc ccggtattaa tttacttctc
cgccttatct tgcaatcttg caatctcttt 180ctcctcgtta tcacgcgttc ctgcaggcgc
acctccgatg gcactgcagc cggagtcccc 240gcggcgccgg cactactaaa gactaaagtg
tctagtctag cctccaatgt gctcacctcc 300atcagcatct catccattta tcttctgacg
atgtcatctg caggctccac cccctccggc 360cgccccgacg ctctccgacg gtgcacaaca
atcaattctg cagtcacgct caagattcgt 420ccctgccgga ctcctcatgc cgtgcctggt
ttaatctatg caatggagta aggtagtatc 480gcctagcagg agcggagttc ctgctgcgct
cacgccatgg tgccggcgca gacataaatc 540gctcgtttcc tccggcgctg gccgttctct
cgagccagtt tgtctgttgt ggttgtagga 600tcctctgttc ccctcgacag ctcacaatgc
gttccttctc cgttgtcgct gccgcgtcac 660tggcgctctc ttgggcgtct ctggcccagg
ctgctcgccc ccgtcttgtg cccaagccta 720tctctcggcc agcttcgagt aagtcggctg
cgactacggg tgaggcttat tttgagcagc 780tgctggacca tcacaacccg gagaagggaa
cgttttccca gcggtactgg tggagtactg 840aatactgggg tggacctggg tcaccggtgc
gtctctgaca tttggtctta tgaccggcca 900tattgaaact tagccggtgg caaggtccgc
aatcatgagg aacattgctg attaaactag 960gtggtcctct ttaaccctgg agaggtctct
gccgatggct atgaggggta tctcaccaac 1020gatactctca ctggtgtcta tgcgcaggag
atccagggtg ccgtcattct cattgaacgt 1080gagtgtcact gctaccatgg aaaaaagaca
ttcgctgatc gaccccaatc tagaccgcta 1140ctggggcgac tcttcgcctt atgaggtgct
caatgccgaa acacttcagt atctcacact 1200ggatcagtcc attctggaca tgacctactt
cgccgagacg gtaaagctgc agttcgataa 1260tagcagccgc agcaatgcgc agaatgctgt
atgttacctt caccgctcta tgtttctgat 1320aggtactgac aacgtagccc tgggtcatgg
tcggtggctc atacagcggt gccttgacgg 1380cttggaccga gtctatcgcg cctggaacgt
tctgggctta ccatgccacc agtgcgcctg 1440tggaggctat ctatgacttt gtaggtgtag
cctgctcttg ttatctatac ttgcagctaa 1500ccaagccagt ggcaatactt ctaccccatt
cagcaaggta tggcacagaa ctgcagcaag 1560gatgtgtctc tggtagccga gtatgtcgac
aaaattggga agaatggaac tgccaaggaa 1620cagcaggagc tcaaagaatt gtttggtctg
ggagctgttg agcattacga tgactttgcc 1680gcgtgagtac ttcaaagtct atagacgagc
ttttctgaca ggaacagtgt cctgcccaac 1740ggaccgtacc tctggcaaga caacgacttt
gtcacaggat actcttcctt cttccagttc 1800tgtgatgctg tcgaggtgag ttaccaccag
attcctcttg attgaagcaa tatactaacg 1860gacacagggt gtcgaagccg gcgcggcagt
gacccccggc cccgagggcg tcggacttga 1920aaaggccctg gccaactacg caaactggtt
caattcaacc atactcccta actgtatttc 1980accatctctt gtctcgttcc tctcccttat
cctcccagac taacctagtg acagactgcg 2040caagctacgg ctactggacc gacgaatgga
gcgtcgcctg tttcgacagc tataatgcct 2100cgagccccat cttcaccgac acctccgtgg
gtaaccctgt cgaccgccaa tgggaatggt 2160tcctctgcaa cgagcctttc ttctggtggc
aggagtgcgt accccttacc tcattcatga 2220taacacacga acaattccac taacaaagat
ccagcggtgc ccccgaggga acctccacta 2280ttgtgccccg gctcgtcagc gcctcctact
ggcaacgcca atgcccgctc tacttccccg 2340aagttaacgg ctacacgtac ggcagcgcga
agggtaaaaa ctccgctacg gtgaacagct 2400ggacgggtgg atgggatatg acccgcaaca
cgacgcggtt gatctggacg aacgggtagg 2460tctcccccta atttccgttg aatgtgatgt
gaagataaac tcaatgctaa taaattgaga 2520aggcaatatg acccctggcg cgactccggt
gtgtcgagca ctttccggcc cggtggtccg 2580ctggttagca cggcgaacga acccgtgcag
attattccgg gcgggttcca ttgctcggac 2640ttgtatatgg aggattacta tgcgaatgag
ggtgtgagga aggtggttga taatgaggtg 2700aagcagatta aggagtgggt ggaggagtat
tatgcttgat gaagatactg gtggacatat 2760ggagtgtaca taagatgaat ggtcataaaa
tgatgatggt agatacggct atggctgttg 2820attagatggt cctttcgcat ttcctaatta
ctgagcacgt gctccatggt atgggaagtg 2880gagacgttgc tatatatatt gactgtcggg
ctattgttca cggcgtagaa gctagacgct 2940ttgtctatgt ggccttcact aaagaccgtg
actctgccca gtcttccccc cttcgaggac 3000ctggtattag ccaaacccac ccacaaacct
aacaaagatc atcgtgacat tgaagtcact 3060ctaggtactg ctggcgctga ttacagtggc
tcaattcgaa catttcaaca gcacataagg 3120gaagggtcgc ttcacttgct accttgatac
gaaagcagcc acgcccaaca cttatagggg 3180tgacaaccat cggcatgctg ggttatctac
tatatctcct gattctgtgg atcctggaga 3240tcgatctggt acactaatct actacaatgc
atgtgaagta gggataggca 3290562044DNAAspergillus niger
56ggctttgttg ggctgagcgc tacttctttc tctctcttgg tctgttcgtt gctccgccag
60ttggttcact cagcctcgta acatcagtat accaggctaa gtcaggactt tggcccccat
120actgcttccc ctttttttat aaaactcaat ccttctggaa aggattctat ttctcaattc
180tcagactact taatacgttc tttgttttca aattgttttg tttctgaaac ttgccgggcc
240ctatcccctc ttttttatag tccgcctgtc gacatcatat ccagagtgag ccaccatgca
300gctcctccag tccctcattg ttgccgtttg cttcagctac ggcgtcctct ccttacccca
360tggcccgtca aaccagcaca aagcacgttc cttcaaggtt gaacgggtcc gtcgtggaac
420cggtgctctg catgggcccg ctgctctccg caaagcatac cggaagtacg gaatagctcc
480cagcagtttc aacatcgatc tggcagactt taaacccatt acgacaaccc atgctgctgc
540tgggagcgag attgcagagc ctgatcagac tggcgctgtc agtgctactt ccgtcgagaa
600cgatgccgag ttcgtttcgc ctgttcttat tggcggccag aagatcgtca tgacatttga
660cactggttct tctgacttgt aagtcttgga tgcagctgtt tactctttgg tacagtgatt
720aacgtcgatc tacagttggg tgttcgatac gaatctcaat gaaaccttga cgggacacac
780ggagtacaac ccttcgaact cctcgacctt caagaagatg gacggataca ccttcgatgt
840ctcgtatggt gacgactcgt acgcctctgg ccccgtcgga acggataccg tcaacattgg
900cggcgccatt gtcaaggagc aagccttcgg tgtccccgac caggtatccc agtcgttcat
960cgaggacacg aactccaacg gcctggtcgg gttgggcttt tcctccatca acaccatcaa
1020accggaggcg caagacacgt tcttcgccaa tgtcgcacca agtctggacg agcccgtcat
1080gaccgcctcg ctcaaggctg acggagtggg cgagtacgag ttcggcacga tcgacaaaga
1140caagtaccag ggcaacattg ccaacatcag cgtggactca tcgaacggat actggcagtt
1200ctccactccc aagtactccg tggcagacgg agagctgaag gacattggaa gcttgaacac
1260ctcgatcgcg gacaccggta cctcccttat gctgctggat gaagacgtgg ttactgccta
1320ctatgcgcaa gttcccaact cggtctacgt gagcagtgcc ggtggttaca tctacccctg
1380caacaccact cttcccagct tctcgcttgt cctcggcgag tcgagcctgg ccacgatccc
1440cggtaacctg atcaatttct ccaaggttgg caccaacacc accaccggac aggcctgtaa
1500gttgctcccc ttcttttgca tgattgaaca tgattgactg attgtgctgg ttagtgtgct
1560ttggcggcat tcaatccaac ggaaacacct cgctgcagat tctgggcgat attttcctga
1620aggccttttt cgttgtcttc gacatgcgcg gcccctcgct tggtgttgcc tctcccaaga
1680actagtttcc ttttcctgta cttttccccc gcgtgtaata atatcgtctg attttttgga
1740ctgtctccta cgtgggcaag atggatggat agtttgctca cgtgcattgc tttaccttgg
1800gtctgtgagt caaggcagga gtgcgtggct gtatctacaa ttcaagttac agtgccgacc
1860gttattgcct tccacatcga aaaacataga cactctttct aaccctaatc catgatacaa
1920gtatatactt cgagtccata ttatggtggt gtatcaaggc gccatgttta tatctaatga
1980aaccaacgta ggtctcatct tcatacgttg tttaaaaggt gccgaagaat atacgaagat
2040agat
2044573916DNAAspergillus niger 57ttcgcagata ttcgagtcaa taccttgtat
attaacggaa aagggtttgg gaagcgttgc 60cttcaaggat gaaattcaaa acgtagagta
tcttgacctc aatgacctca tgatcccctt 120gaccgtacag ctagtgatct cacggtggta
attgctcgat tcgattccaa actacagcaa 180gaagcatgcc cgacaggaaa gtcaagttac
aagagatcta gaacggggct gctacggctc 240tgaccaacct ttgtctgttc tagatgaggc
tgcagtgacg taactccggc ccaagaaaat 300gggtggtaac cgaccgttgg aaatcccatg
gaactggcgg ggttcaaatt cccgataact 360gtaccgaatc cttgtatccg gatataggcg
accgatagta gtagctatag tcagacgtag 420cacagcatta ctgtgtcccc ctggtcataa
gagtatttgt tacaataaga agggaccgtc 480tagttgtgta tcacaacaac tgacacttgc
ggaacaggta tcagatcctt gcgctggcat 540cgactccagc aatggcttgc tctgtttcgg
acgaatatca tcgggctaga ctgtgacctg 600aaggatcgat ccaggagttt gacggcttgg
gtttctaatc cgatctggaa gattattgtt 660tagtcaagac cgggaacagc aagaaccgga
agattatctg tttaactact ttgccaaaac 720agttattcaa tgtcatgctg agtgcgatgg
ggacccaagc tcacgagtga tgctgatcaa 780agggtccgtt cggtttggga aaggaagggc
gccctgtgct tgccgttgga atccacggaa 840cgattccgat tgactgcacg ggatttgcaa
actgactccc agtgggacga agaaagaaac 900ccatcttact ttgcggaggc aggatggcac
tttgtgtcgg aagggtggtg actcctggtg 960ttgccgagaa gatggaatgg tggagaattg
ccttgaaaat gaggctgtag gggcactaac 1020ccagtcgggc cgagcgtgac gcctttcccg
atgaggcagg tcaggtgggt gtcacgggtc 1080tatcttatcg cttatcatct gaccggttcc
tggcagaagc tccccttccc agctcctcgt 1140atttattcct ccccaggttg ttgtttgctc
ttctccctgt ctcgccttct ctccagcctc 1200cattcaacgg gctccatccc cttctctccg
gatccctcta tcccttattc ttccttagat 1260atcattattc atatagtgcc ttgtttcgct
caccatgcgc attgactccg cggcgctaca 1320tctggtccca gtcctcctgg gccaggtcgg
tgctttacaa ttacccttgg tccaagactc 1380caattcacag tggcagaaac caaatgcagg
tgataaaccc ctaattagct ctccgttgct 1440tcaagagcag gtcaaggcgg agaatctgtt
ggacagggcc cggcagcttt acaagattgc 1500ggagctggga gaagacgagt ataaccaccc
cactcgcgtc attggcagta aaggtacgat 1560atattttttc atcatgtccc tggaatacga
taattagctg acaacctttg ctccccaggt 1620caccttggca cgctcgacta catatactcc
acccttaccg acctcggtga ttattatact 1680gtcgtcaatc agtccttccc tgccgtgagc
ggtaatgtct tcgagtctcg ccttgtcctt 1740ggtcacgatg ttcccaagtc agctacacca
atgggtctca ctcccccaac gaggaataag 1800gagccggtat atggctccct ggttgctgta
tccaacctcg ggtgtgaggc ctcggactac 1860tcgtccaact tgaaaggcgc cgttgcattt
atcagtcggg gaagctgtcc gttcgggacc 1920aagtctcaat tagctggtaa agcgggagct
gttgctgccg tcatctacaa caacgagcgg 1980ggtgacctaa gcggaactct aggaaaccca
acccccgatc atgttgctac ctttggtatc 2040tcagacgagg atgctgcccc agtcctggag
aagttgaata aaggcgagaa ggtggacgct 2100atcgcctacg ttgatgcgat agtagagacc
atccacacca ccaatatcat cgcgcagacc 2160acggatggtg acccgaacaa ttgtgtaatg
ctgggtggcc acagtgacag cgtggccgag 2220ggcccgggta tcaatgacga cgggtccggt
actctgaccc ttttggagct tgccacattg 2280ctcacccagt tccgtgtcaa caactgcgtg
cgatttgctt ggtgggccgc cgaggaggaa 2340ggccttctcg gatctgacta ttacgtgtcc
gttctcacac cggaagagaa ccgcaagatc 2400cgcttgttca tggactacga catgctcggc
tcgccgaact ttgcgtacca agtttacaat 2460gccactaatg ctgtgaaccc cgagggatct
gaggagcttc gtgatctgta caccgacttt 2520tacgaagatc atgggttcaa ctacacgtac
attccgtttg acggacgcag cgactatgat 2580gccttcattc ggcatggtat cccgggtggt
ggcattgcca cgggagcaga gggtatcaag 2640actgtcgagg aagcggacat gtttggtggg
gttgctggcc aatggtatga cccgtgttac 2700catcagatct gcgatacggt ggccaatgtg
aacttgactg cgtgggagtg gaacaccaag 2760gtaggaccgc atccagcatt agttactgtt
atgccgatca ttcttttgct aactgggatg 2820atttacctta gctcgttgcc cactccattg
cgacttacgc caagtccttt gacggattcc 2880cggaacggtc cgatgaaccc atcagccctg
ctgcttttga ggaaccgaag taccatggcc 2940acgcgttgca attgtaagat ccgtgtgccc
caaagcttgg tagtggggca aatatctgta 3000gcagagacca agtcctttct cgaacctgag
tcccacttgg tctttccatc ttttgagttg 3060aacaagtaga ccgttactcg tcatgtagac
gaagacctgg cagaaccgtt gtactatgta 3120gacatttatc ggggattgtc tgttatgaat
tcatgtgctt gtgggatatg gttaggtatt 3180tctcgcaaat gaccacggct tcctctatca
tttcccgcct cgcaccctca taccgatttc 3240cgttaactta atatcatctg tagacttcgc
ggtaatacta cagggaccca gagcgtcctg 3300tggggagccc aaatccagaa tggaacagct
gcatcggtgc ttaatctatt gtccatacga 3360cgcagaggca ctttcagtct aagctaaaag
caagcattat tcagagatac agtgagtgtc 3420aatatgccgt tggttcatgc gcgattcgtg
caaaaacagc ccaccgccct atataacaga 3480gtatattgct ctctgcctct ccgggtagag
atcgacattt ccgggaccgc catgaaacgt 3540tggtgggcct cggagcgtcg aggattgaca
ctagggccgc gctagaatcc cccagggtct 3600gcaaaatacc ctatggggtg acctcgaaca
gggcgtgaaa cggcaattcc tccgaaagcg 3660ttgcgaaagt gtctacaaag tggtgatcca
gaccatcacg cggttccgga acaattgccc 3720ttcttatttg gcactttacg gcgtggagtg
gagtcatccc cctttgttga aacaacacgg 3780gcatgtttcc tttgccgtcc ggttcgttac
ctataaagac agggtaggtc tgttcgggaa 3840aaagacacaa tcagcgccta tcccgtccgt
cgaagtctat tgccataccc tatcacggat 3900catcagactc ataaca
3916581443DNAAspergillus niger
58atgcatctcc cacagcgtct cgttacagca gcgtgtcttt gcgccagtgc cacggctttc
60atcccataca ccatcaaact cgatacgtcg gacgacatct cagcccgtga ttcattagct
120cgtcgtttcc tgccagtacc aaaaccaagc gatgctctag cagacgattc cacctcatct
180gccagcgatg agtccctgtc actgaacatc aaaaggattc ccgttcgtcg tgacaatgat
240ttcaagattg tggtagcgga aactccctct tggtctaaca ccgccgctct cgatcaagat
300ggtagcgaca tttcatacat ctctgtcgtc aacattgggt ctgatgagaa atctatgtac
360atgttgctcg acacaggcgg ctctgatacc tgggttttcg gttccaactg cacgtccaca
420ccctgcacga tgcacaatac cttcggttcg gacgattctt cgacccttga aatgacatcg
480gaagagtgga gtgtgggcta tggaactggg tctgtcagcg gcttgctagg aaaagacaag
540ctcacgattg caaatgtcac tgtacgcatg actttcggac ttgcttccaa cgcatcggat
600aacttcgagt cgtacccaat ggacggcatt ctcggtctcg gtcgaaccaa cgatagttcc
660tacgacaacc caacattcat ggatgccgtt gcagaaagta acgttttcaa gtcgaatatc
720gttggcttcg ccctttcacg tagccccgcc aaggatggca cggtcagctt tggcactact
780gacaaggaca agtacaccgg cgatatcacc tacaccgata ccgtcggatc ggacagctat
840tggcgcattc ccgtggacga tgtctatgtt ggcggcactt catgcgattt ctccaacaaa
900tcagccatca tcgataccgg aacttcttat gctatgctgc cttcaagcga ctcgaagacg
960ctgcacagtc tcattcccgg cgccaaatct tcggggagct accacattat tccgtgcaac
1020acaactacta agctacaagt ggcattctct ggtgtgaatt acaccatctc gccgaaggac
1080tacgtgggag caacttcagg ttctggatgc gtttcgaaca ttatcagcta cgacttattt
1140ggtgatgaca tctggctcct gggtgacacg tttctcaaaa atgtgtatgc tgtgtttgac
1200tacgatgagt tacgggtcgg atttgcagag cgttcctcga acaccacctc tgcgtcgaac
1260tctacgagct ctggaacaag cagcacctcg ggatccacta caacgggcag ctcaacgact
1320acgacgagct ctgctagctc tagtagttca tctgatgctg aatcaggaag tagcatgacc
1380attcccgctc ctcagtattt cttctctgct ctggcgattg cttccttcat gctttggctc
1440tag
1443593300DNAAspergillus niger 59atgcttcgtg gtcttcgtga tgtcgtatta
ttacaatttg caatcccctt gttcttgcta 60ttgcatttta gattatcgct acggggtgtg
atcacaggat ttggttctaa atcacatttc 120cagagaccat tgagcaaaat gtcatctact
caaaagagcc atttcaagct actccagaag 180ttcaaaccgg agtactcgcc tagcgagttt
gctcagtatg agtcggagag aacaggcatg 240agggtagtgg tcattgacca aaaaggaccc
aaagtcacag gttattttgt tctagccaca 300gagattctcg atgattcagg tgctcctcac
acgttggagc acttgtgctt tatgggctcg 360cggaactata gatataaggg cttccttgac
aagctagcaa cacgtgttta ttcgagcacc 420aatgcctgga cggccacaga ccacacggcc
tacaccttgg acacagcagg ctgggaaggg 480ttcgctcaaa tcttgcccgt gtacctagag
catgttatag ctccaacact gacagatgaa 540gggtgctata ccgaagtgca tcatattgat
ggcgctggag acgacgctgg agtcgtctac 600tcggagatgc agggtgtgca gaataactct
gcagagttaa tcgatctaac cgctcgtcga 660ttgacttacc cgcatggtgt aggttttcgc
tacgagacag gcggtatgat ggagcagctc 720cgcgtcctca ccgcggaccg tatccgagcg
ttccatcgtg agatgtacca gcccaagaac 780ttatgcctaa tcatcacagg cgaagtagat
caccagaaca tgctggagac cttggacaag 840ttcgaagata ctattctaga tgtcattccc
agtcctgatt cacctttcaa gaggccgtgg 900gtagattcca agcaggcgcc gccattggag
aagtccattg tccagactgt ggaatttccg 960gaagaagatg aatctttcgg ggagatagaa
attagattcc tcggtccgga ctgtaccgac 1020cctgttcaaa ccggggctgt caatgttgca
ttgctgtatc tggccggttc atctgcttct 1080ctattggata acatcctggt tgagaaggag
cagctcgcca gtgctgtcta ttatgctacc 1140gaagatcatc ccagcattga gatccgcttc
acattaacca gtgtggagac agagaaactc 1200gcgaaggtag agcaacggtt tttcgaagtg
ctcaaggacg ctatggagaa agatttagac 1260atgaggtata tcaaggagtg cattgaccgg
caaagacgga cctggaagtt ctctaccgaa 1320agctccgcct cttcctttgc ggagtacgtg
atctcggatt ttcttttcgg aaagagagac 1380ggatcgacta tgcttgatgt tgcgaccttg
caagagtacg acgtgctgga gaagtggagt 1440gaagaacagt ggcgcagttt tatcaaaaca
tggatttctg atgccaacca tgtcactatc 1500cttggtgttc cgtccgttaa gatgtctgac
acattaaaga aggaggagga agctagagtc 1560gcagagcaaa agaagcgctt gggtgatgag
gggctgaaga agttggccga caagctggaa 1620aaagctaaag ctgaaaatga caaggagatc
cccaaggaga tgctggagag gttccaaatc 1680cctggaatag agtctatcca tttcgtggac
actactacag ccaggtctgg tgcagccctc 1740gatgccgggc gcccatccca caaggcgcaa
aaactggtgg atgctgatgg ctctgatctg 1800cccttgttca tccatttcga gcatatcccc
agtagcttcg tgcagctctc cctcctcatc 1860tcggcacagg ccgtacctgt gcagcttcgt
ccactgctgt ctgtgtatac tgaggcattc 1920ttcaacctgc ctgtcaaccg gaacggggaa
accatcaact ttgagcaggt ggttgtcgag 1980ttggaaaggg atactgttgg ctactccatg
gaaggagcta gaagcctagg aaactcggag 2040atgttgcgga tctcattcca ggtggagctt
gagaagtatc acacggcgat cgcatggatc 2100caggaacttt cctggaactc gattttcgat
gtcgagcgac tccgagcgat taccagtcga 2160ctgctctccg atgtgcccga ttccaagcgt
agtggcgacg acatgctcgc ggctgttcat 2220gtgatggtcc actatgcagc agagtctatt
gttcgggctc ggagcacctt ggtgaaggcg 2280cgttatttga aacggatcaa gaagcaatta
gcagaagagc cgaagtctgt cgttgcgcgg 2340atggaagaaa tcagagatgc gcttttccgt
ttcgagaaca tgcgagtctt agttatcgct 2400gacctggaga aacttcaaaa ccctgtgtca
gcatggaaac catttgctga gcgtttgggt 2460gcaggtgccc ctctacagcc tatcacgact
agaagaccgt tgctcagtga ggcaggccag 2520aagttgggcg gtaagtcgta tgtggttcct
atgccgacga ttgattcatc gttcgcatat 2580gctaccgcac gtggtttgga ttcttatgat
gatccaagac ttcctgcctt aatggttgca 2640attgcataca tgaacgcggt tgagggtccc
ctctgggttg cagttcgagg caagggtttg 2700gcatatggca cgaactttgc ctataacatt
gataccggat tcgtcaactt cgacgtttac 2760cgctccccca acgcccataa agccttcgac
tccagcaagc agattgttga ggatcacctc 2820tctggtgcga tgcccttcga tcccttgatg
ctggagggtt ccattagcag cattgtggta 2880agctttgcga atgaacagtc gacaattggt
agcgcagcct caggcagttt catccgacag 2940gtgattcggc gcctgcctag cgactacaag
gagcgggtgc tcaagcaggt gcgggctact 3000agcgttgatg acgtgaaagg cgctctgaag
gacatcattc tgcctttgtt taacccgtcc 3060acggccaata tcgtggttac ctgcgctaca
gtgcttgagg agactatcaa ggaaggtctc 3120caggcatcgg gattcacgcc tgcggtgcag
ccactcaaag aattcgaaga tgactatggg 3180ctgaaggtcg gcgatgacga ggacgaggag
tccgacgatg acgacgatga gtatgaaacc 3240ggatctgaag atgaagatga cagtgatgaa
gacatggagg atgacgaaga tgatgagtga 3300602181DNAAspergillus niger
60atgggagctc ttcagtggct gtccatcacg gctgctgcgg cctccgcagt gtcagccttg
60accccggagc agatgatcgg tgccccacgg agaaccgaag ttataccaaa cccctccggt
120gacaccggtc tattctcgac ctcccaatgg tcgtttgaca ctcattctga gagcacctgg
180tggagcttga tcgacctcca atcgggcaag accaccactc tcaccgatga tagcgatatc
240gaggagatca tctggcttgg ctctgacaat tctacgctcc tctacatcaa cagcaccaac
300gcgcaggttc ccggtggcgt ggagctgtgg attgcggact cttctgactt tgcaaatgct
360tacaaggcag cctctctctc cgccggtttt ctcggcatca aatcaaccgt gacagattcc
420ggcgacgtgc atttcatcct tcgtggaaag tcctatccca acggaacggc atacaatgat
480cagctcgccg agacctatcc cagtacagcc cgcatctacg acagcatctt tgtgcggcac
540tgggacactt acctgaccac cgcctcccac gctgtattct ccggtaccct gcaaagctcg
600accagcgacg acggcaatgt tcaatatacc tcttcagggg gattgaccaa cctggttaac
660ccagtcaagg gtgccgaaag cccattccct ccttttggag gcaacgacga ctatgacctc
720tcgcctgacg gcaaatgggt taccttcaag agcaaagcgc cagagctgcc tcttgctaac
780aacacggctg cctatgtcta tctcgtccca cacgacggct ctgcgactgc ctttgccgtc
840aacggccctg atagtcctgc aaccccggag ggagttgaag gagaatccaa caatcccgtg
900ttctcccctg atagcgacaa aatagcgtac ttccaaatgg ctactaatac atacgagtcg
960gaccgcaacg tgctatacgt atactccatc gccgatgaca ccatcactcc ccttgcaaag
1020gactgggacc gatcgcctag ctccgtgaca tgggtcgatg gagacaacct cgtcgtggca
1080agccaagatc taggacgaac cagacttttc gccatcccag gcgatgcagg ggacgacttc
1140aagcccacga acttcaccga cggcggctcc gtgtcggctc aatacgtcct atccaactct
1200accctcctcg tcacgtccag cgccttctgg acaagctgga gcgtctacac cgccagccct
1260gacgagggcg tgatcaacac actggcctca gccaacgaga tcgaccccga gcttagcggc
1320cttagttcct ccgactttga agagttctac tttgacggca actggactac cctccaagga
1380tggatcacct acccccaaga cttcgactca tccaagaaat accccctcgc cttcctcatc
1440cacggcggcc ccgaagacgc ctgggcggat gaatggaacc tgaaatggca ctccaaggtc
1500ttcgccgacc agggatacgt cgtcgtccag ccaaacccca caggaagcac cgggttcggc
1560cagcagctca cagacgctat ccaacttaac tggaccggcg ccgcctacga cgacctaacc
1620aaagcctggc aatacgtgca cgatacctac gacttcatcg acacggacaa cggcgtcgcc
1680gcgggtccca gcttcggcgc gttcatgatc acctggatcc agggcgatga ctttggacgc
1740aagttcaagg cgctggttag ccatgatggt ccgttcattg gcgatgcgtg ggtcgagacg
1800gatgagttat ggtttgttga gcatgagttc aacggcacct tctggcaagc gcgcgacgca
1860ttccacaaca cggatccatc cggccccagc cgcgtcctcg catacagcac cccccagctc
1920gtcatccaca gtgacaagga ttatcgcata cctgtggcaa atgggattgg actgtttaat
1980acgctgcagg agaggggcgt gcccagtcgg tttttgaatt tcccggatga ggatcattgg
2040gtcaccgggc aagaaaacag cctcgtctgg tatcagcagg tgctgggatg gattaatcgg
2100tattctgggg tgggagggtc gaatcctgat gcgattgctt tggaggatac ggtgaatccg
2160gtggtggatt tgaatccttg a
2181611695DNAAspergillus niger 61atgacgaggc agacttctct cgttcccagg
ctactaacgc tagcctcact agctgcactt 60tcacaagcag agctaggcaa gatccaatgg
aaaggatctt gcaacttgac cacttatccg 120gcattgatct gtggaacact agacgtgcca
tacgactaca cggagtcaaa ttccagcaag 180acactgactc tcgacatcgc caagtggcca
gcgaccaaga aaccagtctc ggagcccatc 240atatttaact ttggaggacc tggtgtcaat
tcgttcgagg gccttgggct ttatggagag 300gaatttcagg ctattcttgg aggtcacaat
gatttgatag cttttaacaa ccgaggcgtt 360ggaaacacca tcccgttctc ctgctacagc
gatgacgcca cccgtgaact cgtcgccctt 420caagctccta acgacggcag agcgtccagc
acggctttgg gagaaatctg ggcccagaac 480gcaaacatcg cacaggcatg ctatgctacg
aacaatcaaa ctggtagtct tattggaact 540agctttgctg caagggacat catgcaggtc
gctgatgcgc tcagtggaaa ggatagtttg 600gtcaactact ggggattctc atacggcact
acaatcggtg ctgttctcgc agccatgttc 660ccggatcgaa tggggaatgt cgcgcttgac
ggagtggaca accccagaga ggctctttat 720ggatacaacg cacaagcggt tgtggacgtc
gacaaagttt tcgaaggatt ctgcacgggc 780tgcatggccg caccggacct ctgccctatc
gccaaggagt acaccagcgc cgccaacttg 840gaagccgcaa tttacctgat gctggaaaac
ctcaagtaca acccgattgc cattcccgaa 900accggtggaa tcgtaacttg gagcgacgtc
aagtcgacca tttttgaggc catgtacctg 960ccaagctctt ggcccttgac ctctgagctt
ctttactacg tgcaaacccg caacacaacg 1020atccttggca actctgaagt atacgacacc
atcaaatcct acggtcaatc ggcttctttg 1080acttcggctt ccgatgaggt cggcacggcc
attacatgct ccgacaagca tcgatctgcc 1140accattaaag aggtcctccc gtacgtcaaa
gccagacagg ctctgaccaa gatcggaagt 1200gatggctcgg acggcgacat gagatgcgcg
cagtggaatc cgaagatgtt cgccaaggag 1260cgctactccg gtgactttga agtcaagaca
gccaacccgg tgttgattct gagcaacact 1320tacgatccag cgactcctct tcccgcagcg
aagaacctga cagagacctt tgagggaagt 1380gtcttgctcg agcagaacgg atacggtcat
actaccctgt ctatgccatc tctttgcact 1440gccaaggccg tccgggctta cttcaccaat
ggcacattgc ccgctgacgg aacgatctgc 1500caggtggacg tgcctctgtt cacgaacttg
acctacaagg atgtgtggcc gaagagtttc 1560caacggagcg ttgagtcgag ggatgatgcg
actatcctca aggctttgat gtcggtccgt 1620gataagatgt cgcgacgcag gatgtgtatt
tatttgtaca ccaacagcgc ttcatggaga 1680ccggaacttc cctga
1695621581DNAAspergillus niger
62atgtactact ctctctgggt tgctgccttg gtggccgcgc tgcccgtctc ccgggcccag
60tttgtggctc cgcccacgga tctcattccc accaagggat atctcgacat ccccgtccgc
120tacaaacagg tccccaccgg catttgtgag actgatccca gtgtcaagag cttctccggt
180tacgtcgatg tcgctgagca tgagcacatc ttcttctggt tcttcgaggc gcgcaaccaa
240gatcccaccg aggctccctt gaccgtctgg atcaatggag gcatgtctga ccccggtcct
300ggttcctcct ccatgatcgg cttgttccaa gagcacggcc catgcggcat tgacgccaat
360ggctccgtct acaacaaccc ctactcctgg aacaacgcca gcaacatgct ctacatcgac
420cagcccgtgc agaccggctt ctcctacagc attccggttc ccggctatgt ggattcttcc
480acagacaatg gttttatggg cgcatttcct cagtactcgc gcgaaacctt ccacttcacc
540acggagagtt atggcggcca ctacgggccc gtcttcaacg agtacatcga ggagcagaac
600gcccatctcc agccgggagc caagaagatc caactgggca gtgtgatgat cggcaatggc
660tggtatgacc cgattattca ataccaggcc tactacaact ttacggtata tccgggcaac
720acatacgact acctgccatt caacaagtcc atcagctcgc tgatgtacaa caacctctat
780ggccccggaa actgcctcga ccagctctac gactgcgccg cccgaggcat cgacgagatc
840tgcagcactg ccgacgattt ttgcgccaac gaggtcgaaa acgtctacga catttactcc
900ggtcgggatg agtatgactt tcgtgaactc actccggacc cgttccctta cgagttctac
960gttgactacc tgaacaaagc gtccgtgcag gccgccatcg gcgcatacat caattacacg
1020gagagcaaca acgctgttgg actcgccttt tcgtccaccg gtgacgacgg gcgactcatg
1080aacaccatcc aggatgtggg caagctgctc aaacagggtg tcacggtggt catgtacgcc
1140ggggatgccg actataactg caactggctg ggtggggaag ccgtgtcgtt gcaggtcaag
1200gccgccaact tcagtagtgc gggttacacc aacattgtca cctcggatgg agtgacacac
1260ggccaggtgc gccaggcggg gcaatttgcc tttgtgcgag tgtatgagag tggacatgag
1320gttcccttct atcaaccctt gcttgcgctg gagatgtttg agcgcgtcat tggcggcaag
1380gatgtggcga cgggaaagat tcccatctcg tcgagtttac agacggtggg cacgcccaag
1440agttactacc gggagggcaa cagcacgatt cagtgggagg tgttggattc tctggcgacg
1500tacaacacaa ccacgaatgc tccgaacccg gtgagccgga ggctgaagcg gatgggacca
1560gctttgcggt ttcagatgta g
1581633471DNAAspergillus niger 63atgtcttgcg tctggctcca catccacaaa
aggagcctac tgtctgtcgc tacgaacaat 60tctgttgcga gggccgctgc ctctacctcc
gccgcgccgc cgccgccgtc atcgccgccg 120cctggttcta atacttattc gcctctttat
cgccccatca ccaatcccat cggatttact 180ttgtcgcctg cgaggtcact agtttctcgc
aatcctaaat ttcctgccta tcggcgctct 240agtcgacact tttctttgtg cccggccgct
gcaacgcccg gtgtcaccac gagcatctgc 300cctggtcagg cgcccgtccg ctctctcagc
tcgctcatta tacactctac gagaccccgt 360gctatacgta tccgtaccga ccagatggat
ttgaatggag acgcaggcgc caagcgcaag 420cgcagctcca tcaccacacc cgccgaacgg
cccgtaaagc accttcgccc cgaatcgagc 480gcattgacac ccggggattc gacgcctgcc
aatgggactg tatacgatgt ggaggatgat 540gaagatgcga gtcgtctgct gcctgtaggg
cctgctcagg ccgactcacc ggaatggcaa 600gctaccatag aggaggttgt gaaaagcgta
gtgtctatcc acttctgtca gacctgctcc 660ttcgacacgg agctgtccat gagtagtcag
gctactgggt ttgtggtaga tgcagagaat 720gggtacatat tgacaaaccg acacgtggtt
tgcccgggac ctttctgggg atactgcatc 780tttgataacc atgaggaatg cgacgttcgt
cctgtgtatc gggaccctgt tcacgacttt 840ggaattttga aattcgaccc gaaggctatt
cgatatatga aattgaggga actgaaactg 900cagccggatg cagctaaagt gggatcagaa
attcgcgttg tgggtaatga tgcaggagaa 960aaactgagta ttctgtctgg tgtcattagt
cggctggata gaaacgcgcc cgaatacggc 1020gatggctaca gtgacttcaa tacgaattac
atccaggccg ccgcagcagc tagcggtgga 1080agttccggca gtcctgtagt taacattgat
ggccatgcga ttgctctgca ggccggtggt 1140cgtgcagacg gtgcagcgac ggattacttc
ctccctctgg accgaccgct acgcgcactg 1200gaatgcatcc gtcgcggaga gcctgtcacg
cgtggaacga ttcagacgca gtggatcttg 1260aagccgttcg acgagtgtcg tcggttgggc
ttgacgcctg agtgggaggc gaccgtgcgt 1320aaagcagcgc ccacggaaac cagcatgctg
gtggccgaga tcatcctgcc tgaaggcccg 1380gcggacggaa agctcgagga aggagacgtg
ctcctgcagg tcaacggggt gcttctcacc 1440caattcatcc ggttggatga catcctggat
tcgagtgttg ggcagacagt gcgtctgctt 1500gtccaaagag gcggtcagaa tgtggagatt
gagtgccagg ttggcgacct gcatgccatc 1560acgcccgacc ggttcgtgac ggtggctgga
ggcacgttcc ataacctgtc ttaccagcag 1620tcgcggctgt atgccatcgc tactcgcggt
gtctacgtct gcgaggctgc cggctccttc 1680aaactggaaa acacactgtc aggatggatc
atcgactcgg tggacaagcg gcccactcgc 1740aatctggatg agttcgtgga ggtgatgcga
acgattcccg atcgttcgcg cgtggtcatc 1800tcgtatcggc atattcgcga tctccacacc
cgaggcacca gcatcgtcta tatcgatcga 1860cactggcacc ccaagatgcg actggctgtg
cgcaacgacg acaccggtct gtgggacttt 1920tcggacctcg cggaccctat cccagctctt
cctccggttc cgaggaaagc cgatttcatt 1980caactcgatg gtgttagcca gcctgctgcg
gccgacattg tgcgcagctt cgtacgagta 2040tcctgtacga tgcccctgaa gctggacggc
tacccccagg ccaagaagac tgggttcgga 2100ttggtcgtcg atgcagagaa gggtttggtg
gttgtgtcgc gagcgatcgt gccgtacgac 2160ctctgcgaca tcaacgtcac ggtggccgac
tccatcatcg tgaacgctaa agtagttttc 2220ctgcatccgc tccaaaacta cagcatcatc
cagtacgacc caagcctggt gcaggcgccg 2280gttcagagtg ccaaactcgc caccgactac
atcaagcagg gacaggacac gatctttgtg 2340ggattcaacc agaacttccg gattgtcgtg
gccaagaccg ccgtaaccga catcaccact 2400gtttctattc cagccaacgc gtccgcaccg
cgctaccgcg cgatcaacct ggacgccatc 2460actgtggaca ccggactcag cgggcagtgt
tctaacggtg tcctgattgg cgaggacgga 2520gtggtgcagg cattgtggtt gaactatctt
ggagaacgca catctaattc gcataaggat 2580gtggaatacc atctaggatt tgcgactcca
tctcttcttc ctgtcctgtc gaaggtgcag 2640cagggagaga tgccggaatt gcggattctg
aacatggaga gctacgtggt ccagatgagt 2700caagctcgta tcatgggcgt gtcggaggaa
tggatcgaga aggtgacgca agctaaccca 2760tcgcggcatc agctcttcat ggtgcgcaag
gtcgattgcc caccgcctgg gttcaactca 2820gcggccgaca cgttcgagga gggtgatatc
atcctgacct tggacggaca gctgatcacc 2880cgcgtctcgg agttggatat catgtacgag
aaggatacgc tggaagccct gattgttcga 2940aatggacaag aaatgcggat ccaggtgccg
actgttccaa cagaggacct agagactgac 3000cgtgcggtcg tgttctgtgg tgctgtgttg
cagaaaccac accatgcggt ccgtcagcag 3060atttctaagc tacacagcga agtctacgtc
agcgcaagaa gtcgcggatc cccctcctac 3120caatacggct tggccccaac caatttcatc
accgccgtaa acggcgttcc aaccccgaac 3180ctggaccgct tctccgaaga agtgagcaaa
atccccgaca acacatattt ccgcctacgg 3240gcggtgacat tcgacaatgt gccgtgggta
gtgaccgtga agaagaacga tcattacttc 3300cccatgtccg agtatatcaa agaccagtcc
cagccttccg gttggcggac cgtgtctcac 3360gacaaggata aatataaaga cggcattgca
ccggatgctg cgaacttgaa cccggatgct 3420atggacgaag ggtttgatgg agtcagtgat
attgagccgg atttggagtg a 3471641611DNAAspergillus niger
64atgagagtcc ttccagctgc tatgctggtt ggagcggcca cggcggccgt tcctcccttc
60cagcaggtcc ttggaggtaa cggtgccaag cacggtgccg accatgcggc cgaggtccct
120gcggatcaca gtgccgacgg gttctccaag ccgctgcacg cattccagga ggaactgaag
180tctctctctg acgaggctcg taagctttgg gatgaggtgg ccagcttctt cccggagagc
240atggatcaga accctctctt ttccctcccc aagaagcaca accgccgtcc cgactcgcac
300tgggaccaca tcgtcgatgg caagctggaa gcctatgatc tcagggtcaa gaagaccgat
360cctggctctc ttggcatcga ccccggcgtg aagcagtaca ccggttatct cgatgacaac
420gagaatgata agcatttgtt ctactggttc ttcgagtctc gcaatgaccc cgagaatgat
480cccgttgttc tgtggctgaa cggtggccct gggtgctctt ccctcaccgg tctcttcatg
540gagcttggcc ctagcagcat caacaagaag atccagccgg tctacaatga ctacgcttgg
600aactccaacg cgtccgtgat cttccttgac cagcctgtca atgtcggtta ctcctacagt
660aactctgctg tcagcgacac ggtcgctgct ggcaaggacg tctatgcctt gcttaccctc
720ttcttcaaac aattccccga gtatgctaag caggacttcc acattgccgg tgaatcttat
780gctggtcact atatccccgt cttcgcttcg gagatcctgt ctcacaagaa gcgcaacatc
840aacctgcagt ccgttctcat tggcaacggt ctcaccgacg gatacaccca gtacgagtac
900taccgtccca tggcctgcgg tgacggcggt tacccagctg tcttggacga gagctcctgc
960cagtccatgg acaacgctct tcctcgctgc cagtctatga ttgagtcttg ctacagttcc
1020gagagcgctt gggtttgtgt cccggcctcc atctactgta acaacgccct ccttgcccct
1080taccagcgca ctgggcagaa cgtctatgat gtccgtggta agtgcgagga tagctctaac
1140ctttgctact cggctatggg ctacgtcagc gactacctga acaagcccga agtcatcgag
1200gctgttggcg ctgaggtcaa cggctacgac tcgtgcaact ttgacatcaa ccgcaacttc
1260ctcttccacg gtgactggat gaagccctac caccgcctcg ttccgggact cctggagcag
1320atccctgtct tgatctatgc cggtgatgct gatttcattt gcaactggct gggcaacaag
1380gcctggactg aagccctgga gtggcccgga caggctgaat atgcctccgc tgagctggag
1440gatctggtca ttgtcgacaa tgagcacacg ggcaagaaga ttggccaggt taagtcccat
1500ggcaacttca ccttcatgcg tctctatggt ggtggccaca tggtcccgat ggaccagccc
1560gagtcgagtc tcgagttctt caaccgctgg ttgggaggtg aatggttcta a
161165840DNAAspergillus niger 65atgaagttca caaattatct cttgacgact
gcaacgctcg caagcagtgt cctagcggct 60cctgctcccc gcaccggttt ggaggacaga
ctccgtgccc ggtcattgca gcgtcaatca 120catcctctgg cacctattcc acttgacaca
tccaccaaag agaattccag actcctcgaa 180gccgacgaga ataccaccca tgttacatac
agcagtaact gggcgggcgc agtgcgcgag 240caaccacctc cgcaaggcac gtattctgcc
gtgtcggcaa cctttcgtgt accagaaccc 300acggcgcaag gggggagcgg aacgcaggct
gggtcggcct gggtcgggat agatggcgac 360acatacagca acgccattct acagacagga
gtcgacttct acgtggaaaa cgggcagacg 420tacaacgatg cctggtatga gtggtaccca
gactatgcat atgacttcga cctagatgta 480agcacagggg acacgatcgt cgccaaggtg
gaagccatct cgccaagtca aggtgtagcc 540actattgaga acatatcgac ggggaagaag
gccacgcaga cgatcagagc cccagctgcg 600acagctaccc ttgccggcca gaatgccgac
tggatcgtgg aggatttcca gtctggcgac 660tcaatggtcg atctggctgg ctttggcgag
atcagcttct ggggcgtgca agcacaagga 720ggagggtcta catggggtgt agatgatgcg
actattgtcg aactgaagca gggcaacgaa 780gtgttgacag acgtggaggt gcaaagtgat
tcggccttta cggtgaaata tacgagctga 840661722DNAAspergillus niger
66atgatatatg tcaactatat cctgggactt ctgtccctct tacacaccgc tgtagccaca
60gctcctgatt atgtcgtggt agaccaactg aacagcatcc ccgacggatg gacaaaaggc
120gcagctcccc cgccatttac tccgatgaag ttctggttgt cgatgcatca cgagtacaag
180gcggacttcg agcagaaagt catcgatatc tcgacacccg gtcaccggga ttatggacgg
240catatgaaac gcaacgatgt catggccttt atgcgcccat ccgatcaggt ctcaaagatc
300atcttctctt ggcttgagtc ggagcatgtt ccaccaaatg ccatcgaaga tcgcggggat
360tgggtcgcct tcacagtccc gttggcccaa gcacaatcaa tgatgaagac cgatttttac
420aacttccacc acctggaaac aaacacaacc caaattagga ccctcaagta ctccgttccc
480gagcaagtcg atgctcatct gcaaatgatc cagccaacga ctcgcttcgg ccgacctaag
540acacaaacca gcctaccgag cctcatgcca gtgtcggtta acattgatga aataagcgaa
600gactgcttga caggcgtgac gcccatttgc cttcgccagc tctatggttt acctagcacc
660aaggcaagcc ccgactcgag aaacgtcctc ggaatttccg gctatctgga ccagtacgcg
720cgctacagtg acctcgacga gtttctagcc gtatactctc caaacagcgt agacgccgac
780ttctccgtag tatcgatcaa cggaggccaa aacccacaaa actcacaaga gggaagcaca
840gaggccagtc tcgacatcca atacgccctc tccatggcat ttgacgctaa cgcgactttc
900tacactaccg ccggacgtgc gccatccccg tatctcgaac agctccagta tctggtgggt
960cttccggacg aggatcttcc tgcagtgctt agcacgtctt acggcgagga tgagcaaagt
1020ctgccggagg aatacacaga agccacgtgc aatttatttg cccaattagg tgcacgcggg
1080gtctcggtga tcttcagcag cggagactcg ggcgtcggag gatcgtgtgt atctaacgac
1140ggaagccaga ggacccgctt tcagcctatc ttcccggcgt cgtgcccgtt tgttacatcc
1200gtgggtggga ctgagggcgt cgggccggaa aaggctgtgg acttttcgag tggagggttc
1260tccgagcgct ttgctcgccc gtcgtaccag aatgcgagtg tggaagcata ccttgcccgc
1320ttaggagata aatgggatgg attgtataat ccagacggac ggggtattcc tgatgtgtcg
1380gcccaggcta gcaactatgt aatcagggac catgggcaat ggctacaaac tgcgggaaca
1440agtgctgccg cccctgtctt tgcagcagtc atctctcgac tgaacgctgc acgtctcgag
1500cagggtaaac ctacactagg gtttctgaat ccttggctgt actcactcga ccagcaagga
1560tttacggata ttgtagacgg cggatcagtg ggttgtgacg ggtcaaatgg aggagctctt
1620gtcccgtatg ccagttggaa tgccaccaag ggatgggatc cggttactgg gctggggaca
1680cctctgtatc agactctgga gcagttggcg cagtctgctt ag
1722671758DNAAspergillus niger 67atgcgttctt ccggtctcta cacagcactc
ctgtgctccc tggccgcctc gaccaacgcg 60attgtccatg aaaagctcgc cgcggtcccc
tccggctggc atcatgtcga agatgctggc 120tccgaccacc agataagctt gtcgatcgcg
ctggcacgca agaacctcga tcagcttgaa 180tccaagctga aagacttgtc aacacctggc
gaatcgcaat acggccagtg gctggaccag 240gaggatgtcg acacgctgtt cccggtggcc
agcgacaagg ctgtgattaa ctggctgcgc 300agcgccaaca tcacccatat ttcccgccag
ggcagcttgg tgaactttgc gaccacggtc 360gataaggtga acaagcttct caacgccacc
tttgcctact accaaagcgg ctcttcccag 420agattgcgca caacagagta ctccatcccg
gatgatctgg tcgactcaat cgacctcatc 480tccccaacga ccttcttcgg caaggaaaag
accactgctg gtctgaacca gcgggcgcaa 540aagattgaca cccatgtggc caaacgctcc
aacagctcgt cctgtgccga tgtcatcacg 600ctgtcctgcc tgaaggagat gtacaatttt
ggcaactaca ctcccagcgc ctcgtcgggc 660agcaagctgg gcttcggcag cttcctgaac
gaatccgcct cgtattctga ccttgccaag 720ttcgagaagc tgtttaacct gccctcccag
agcttttccg tggagttggt caacggcggt 780gtcaatgatc agaatcaatc gacggcttcc
ttgaccgagg cggacctcga tgtggaattg 840ctcgtcggag ttgctcatcc cctcccggtg
actgagttca tcacttctgg cgaacctgcc 900gccgacaacg agaacgagcc ttacctccag
tactatgagt accttctctc caagcccaac 960tcggctctgc cccaagtgat ttccaactcc
tatggtgacg acgaacagac cgttccagag 1020tactacgcca agcgagtctg caacctgatc
ggacttgttg gcctgcgcgg catcagtgtc 1080ctcgagtcgt ccggtgacga aggtatcgga
tctggctgcc gaaccaccga cggcaccaac 1140cgaacccaat tcaaccccat cttcccggcc
acctgtccct acgtgactgc cgtgggagga 1200acaatgtcct atgcccccga aatcgcctgg
gaagccagtt ccggcggatt cagcaactac 1260ttcgagcggg cgtggttcca gaaggaagct
gtgcagaact acctggcgca ccacatcacc 1320aacgagacca agcagtacta ctcgcaattc
gccaacttta gcggtcgcgg atttcctgac 1380gttgctgccc atagctttga gccttcatat
gaggttatct tctacggcgc ccgctacggc 1440tccggcggta cctcagccgc gtgtcccctt
ttctctgcgc tagtgggcat gctgaacgat 1500gctcgtctgc gggcgggcaa gtccacgctg
ggtttcttga accccctgct ctatagcaag 1560gggtacagag cgttgactga tgtgacgggg
ggccagtcga tcggatgcaa tggcattgat 1620ccgcagaatg atgagactgt tgccggcgcg
ggcattatcc cgtgggcgca ctggaatgcc 1680acggtcggat gggatccggt gactggattg
ggacttcctg actttgagaa gttgaggcag 1740ttggtgctgt cgttgtag
175868798DNAAspergillus niger
68atgaagacta ctgctctctt gaccgccggc ctgctggcca ccactgctat ggccgctcct
60ctgacggcca agcgccaggc tgctcgggcc aagcgctcca cgaaccgcca gagcaaccct
120cccttcaagc ctggcaccaa cgaggtcctc gcccttaacg gcaccaagaa tgtggagtac
180agctccaact gggccggtgc cgtcctcatt ggcactggtt acactgccgt gaccgccgag
240ttcgtcgtgc ccaccccctc cgtgccctcc ggtggctcga gccgcgagga gtactgtgcc
300tccgcctggg tgggcattga cggtgacacc tgtgacactg ctatcctcca gaccggtgtg
360gacttttgtg tccagggcag cgaggtgagc ttcgatgcct ggtacgagtg gtaccccgac
420tacgcctacg acttcagcgg catctccatc tcggccggtg ataccatcaa ggtcaccgtc
480gatgccagca gcgacaccac cggtactgcc acgattgaga acgtgagcac tggtaccacg
540gtcacccaca gcttcacggg cggtgttgat ggtgatctgt gtgagtacaa cgctgagtgg
600atcgtcgagg acttcgagga ggatgactcc ctcgttccct ttgccgactt tggcaccgtg
660actttcacca gctgctccgc taccaaggat ggttcctctg ttggccctga ggatgctacc
720atcatcgaca tcgagcagaa tgaggtgctg acctccgttt ccgtctccag tagcgaggtc
780gttgtcaagt acgtctaa
798691743DNAAspergillus niger 69atggtcgcct tttcccgcat ctcggcaggc
ttcgcccttg ccgcccctgc cctggccagc 60gtcgtcctgg agaccgtcaa gtctgttccc
agcgactgga agctcgtgga ggctgctgat 120accagctcca caatttcttt gtccgttgct
ctggcgcgtc agaacctgga ccagttggag 180gagaagctcc tggccgtgtc cacccctggc
aaggacacct acggccagtt cttggatctg 240gacgacatca atgagcagtt tcctctcgca
gatgacgctg ctgttgtggc ttggctgaag 300aaggcaggcg tcacccagat ccataaggag
ggtggtctgc tgaactttgc gaccactgtg 360ggcacagcca accagcttct caacaccacc
ttctcggtgt acaagagcgg atctacccag 420aagctgcgca caacgcaata ctctgttccg
gatgagctga ccgggtccat tgatctcatc 480tcgccgactg ttttctttgg aaagtccaac
gctgcgcgct cggcggccgt gcgtgcttcg 540cagactacca aggagaccag cagaaagaag
agcagtaatg tgtgcgagta catcactccg 600gattgcctca aagagcagta tagcattgac
tatacgcccg aggcatcgtc gggaagtcgt 660gttgggtttg gcagtttctt gaacgagtcg
gccttgtact cggatttgga tctgttcacc 720cagtactttg acattcccca gcagagtttc
actgttgaga ctatcaacgg gggaatcaac 780aaccaggaga atgatccgga tggtgaagcc
gatctcgatg tccagaacat cgtgggcatc 840tcgcatccct tgccggtgac ggagtacatt
accggaggat ctcctccatt cattcccgac 900gtcgagacta ctaccgacga gaacgagcct
tacctgcagt actacgagta tctgctggcc 960aagaccaacg acgagctgcc actggttatc
agcaactcgt acggcgatga cgaagatacc 1020gttcccattg cctacgccac ccgcgtatgc
aacctcatcg gcctgatggg cacacgtggt 1080atctccatcc tcgagtcttc cggcgactct
ggtgtgggcg gcgcatgcat gtccaacgac 1140ggcaccgaca agaccgaatt cacccccatg
ttcccaggaa catgcccgta catcaccgcg 1200gtcggcggca cccaagacgt gcccgaagtc
gcctgggtgg acagctccgg cggcttcagc 1260aactacttct cgcagccgtc gtaccagtcg
gatcaggtgg agacctacct ggacaagtac 1320atctctgcct cgacgaagaa gtactacgag
cagtacacca acttcagcgg tcgcgcgttc 1380cctgacgtgt ctgcgtttgc aggttctcct
tactacgaaa cttatattga tggtcagctc 1440ggccttgtgg cgggtacttc tggcgctagc
cctgtgtttg cggggatcgt cgcgctgctg 1500aacgatgccc gtctgcgggc caacaagaca
tccttgggct tcctgaaccc ttggctgtac 1560tcgagcggct acaagagcct gaatgacatt
accagtggcg aggcagtggg ctgccaaggc 1620gatgtggagg gcgctggagt cattccttgg
gcgagctgga atgccacgac gggatgggat 1680ccggcgacag ggctgggaac gcctaatttt
gccaagctga aggaggcggt tcttgcgttg 1740taa
1743701896DNAAspergillus niger
70atgcatggtc tgcgcctagt atgcagcata gggacattgc ctttggttat cctggcatat
60ccggcggctt cattgcatac aacttcagca gccgtggact tggactccct tcgtctgacc
120tctaactccg aatacgtcaa ttctgtccat gtagacacga atcgatcagt cgcagtgtcc
180gctgaagaac attataccga tacagcagct cgactggttc agaacattgt tcctggagcg
240agctttcgtc tcatcgatga ccactttgtc ggcgacaatg gagttgcaca tgtatacttc
300cgccaaacgc tccatggtat tgacattgac aatgcggatt tcaatgttaa tattggaaaa
360gatggactgg tcttgtcttt cggacattcg ttcttcacag gcgcgttgcc gagcagccat
420ctggacaata ccaacgtttt gagtccggag gctgcactta gaggagcaag ggacgctata
480cagcttccac tgactattga caatgtttct actgaagctg cagaggggcg gaacgagtac
540atattcagag aggcagtggg agcggtatct gaccccaaag ctaagctagt ctaccttgtc
600aagccagaag ggactctggc gctcacctgg aggatagaaa cagacatgta tgagcactgg
660ctactgacat acattgatgc agagactacc actgtccacg gcgtggttga ctatgtcgca
720gacgcgacat atcaagttta tccctggggc acaaacgatc cagcagaagg acatcgcacc
780attgtcaccg acccctggga cctatccgca tccgcataca cctggataag cgatggacgg
840gacaactaca ccacaaccag aggcaacaat gccatcgcac actggaatcc gaccggcggt
900ggctcctatc tctacaacct acgtccatcc gaccccaact tgaatttcca atggccatac
960tccccaaaca tgtccccacc ccgatcatac atcaacgcct ccatcgtcca actcttctac
1020acagcaaacg cctaccacga cctcctctat acactcggct tcaccgaatc cgctggcaac
1080ttccaatgga ataacagcgc ccacggcggc cgagacaaag actacgtgat cctcaacgca
1140caagacggct ccgggttcag caacgcaaac tttgcaaccc cacccgatgg tatccccggc
1200cgtatgcgca tgtacatctg gatcgagtct actccgtcgc gtgatggaag ttttgacgcg
1260ggcattgtaa ttcacgaata cactcacggt gtatccaatc gtctcaccgg cggctcccac
1320aacgccggat gcctcagcgc cctcgaatcg ggtggcatgg gcgaaggctg gggcgacttt
1380atggcgacgg ccatccgaat caagcccaac gatacacgca caacgtctta cactatgggt
1440gcatgggcag ataatgataa atgtggtgtc cgggactatc cttattctac ctcctttact
1500gagaaccctt tgaactatac gagcgtgaat accatgaacg gcgtgcacgc catcggaact
1560gtctgggcaa ccatgctata cgaggtcttg tggaacctca tcgacaagta cgggaagaat
1620gatgggtcga ggccggtgtt tagaaacggg gtgcctacag atggaaagta cttgatgatg
1680aagttggtgg tggatgggat ggcactgcaa ccatgtaatc cgaacttcgt gcaagccagg
1740gacgcgatcc ttgacgcaga cattgtgttg actggcggga agaatcgctg tgagatctgg
1800agggggtttg cgaagagagg attggggcaa ggagcggctc atagtagttt aaattggatg
1860cggaggggga gtacacttct tcctacggga tgttag
1896711185DNAAspergillus niger 71atggtcgtct tcagcaaaac cgctgccctc
gttctgggtc tgtcctccgc cgtctctgcg 60gcgccggctc ctactcgcaa gggcttcacc
atcaaccaga ttgcccggcc tgccaacaag 120acccgcacca tcaacctgcc aggcatgtac
gcccgttccc tggccaagtt tggcggtacg 180gtgccccaga gcgtgaagga ggctgccagc
aagggtagtg ccgtgaccac gccccagaac 240aatgacgagg agtacctgac tcccgtcact
gtcggaaagt ccaccctcca tctggacttt 300gacaccggat ctgcagatct ctgggtcttc
tcggacgagc tcccttcctc ggagcagacc 360ggtcacgatc tgtacacgcc tagctccagc
gcgaccaagc tgagcggcta cacttgggac 420atctcctacg gtgacggcag ctcggccagc
ggagacgtgt accgggatac tgtcactgtc 480ggcggtgtca ccaccaacaa gcaggctgtt
gaagcagcca gcaagatcag ctccgagttc 540gttcagaaca cggccaatga cggccttttg
ggactggcct ttagctccat caacactgtc 600cagcccaagg cgcagaccac cttcttcgac
accgtcaagt cccagctgga ctctcccctt 660ttcgccgtgc agctgaagca cgacgccccc
ggtgtttacg actttggcta catcgatgac 720tccaagtaca ccggttctat cacctacacg
gatgccgata gctcccaggg ttactggggc 780ttcagcaccg acggctacag tatcggtgac
ggcagctcca gctccagcgg cttcagcgcc 840attgctgaca ccggtaccac cctcatcctc
ctcgatgacg aaatcgtctc cgcctactac 900gagcaggttt ctggcgctca ggagagcgag
gaagccggtg gctacgtttt ctcttgctcg 960accaaccccc ctgacttcac tgtcgtgatt
ggcgactaca aggccgttgt tccgggcaag 1020tacatcaact acgctcccat ctcgactggc
agctccacct gctttggcgg tatccagagc 1080aacagcggtc tgggactgtc catcctgggt
gatgttttct tgaagagcca gtacgtggtc 1140ttcaactctg agggccctaa gctgggattc
gccgctcagg cttag 1185721197DNAAspergillus niger
72atgaagtcag cctccttgct cacagcatcc gtgctgttgg gctgtgcctc cgccgaggtt
60cacaagctca agcttaacaa ggtgcctctg gaagagcagc tttacacgca taacatcgac
120gcccatgtcc gcgctctggg ccagaagtac atgggtatcc gcccgtccat ccacaaagag
180ctggtcgagg agaaccctat caatgacatg agccgtcatg atgttctggt ggacaacttc
240ctgaacgcac agtacttctc tgagatcgag ctgggtactc ccccccagaa gttcaaggtt
300gtcctggaca ctggcagctc gaacctttgg gttccttcga gcgaatgcag ctctatcgcc
360tgctacctcc acaacaagta tgattcgtct gcctccagta cgtatcacaa gaatggcagt
420gaattcgcca tcaagtacgg ctctggcagc cttagcggat tcatttctca ggacaccctg
480aagattggcg acctgaaggt caagggacag gacttcgctg aggcgaccaa tgagcctggc
540cttgcctttg ccttcggccg gttcgatggc attctcggct tgggttatga caccatctcc
600gtgaacaaga ttgttcctcc cttctacaac atgcttgacc agggactcct cgacgagccg
660gtctttgcct tctaccttgg agataccaac aaggagggtg acgagtccgt ggcgaccttc
720ggtggtgtcg acaaggacca ctacaccggc gagctgatca agattcccct ccgtcgcaag
780gcttactggg aggttgagct tgacgccatt gctcttggcg atgatgttgc tgagatggag
840aacaccggtg tcattctgga cactggtacc tccctgattg ctctgcctgc tgacctggct
900gagatgatca atgctcagat cggtgctaag aagggctgga ccggccagta caccgttgac
960tgcgacaagc gctcgtccct gcccgatgtt actttcaccc ttgccggcca caacttcacc
1020atctcctcgt atgactacac cttggaggtg cagggctctt gcgtcagtgc cttcatgggc
1080atggacttcc ctgagccggt tggtcccttg gccattttgg gcgatgcgtt cctgcgcaag
1140tggtacagcg tgtatgacct gggcaacagc gctgttggtc tggccaaggc caagtaa
1197731182DNAAspergillus niger 73atgcgcaagt accgcttcca tcccaccaag
cctggtccct acactctcag cagctccatc 60caacagaccg gtcgtccgta cactgaaaag
cccatcgggg gtcgggccca tatccggcag 120ctggtgcgga agaagagcac caccagcgat
gaggttggcg aggttccggc cgaagatgtg 180cagaacgact ccatgtatct ggcgaccgtg
gggatcggaa ccccggcgca gaacctgaag 240ttggactttg acactggttc agctgatctt
tgggtctggt ccaacaaact cccctcaacc 300cttctatccg agaacaagac ccatgcgatc
ttcgactcgt ccaaatcgag caccttcaag 360accttggaag gtgaatcctg gcaaatctcc
tacggagatg gatcctccgc atcagggagt 420gtgggcaccg acgacgtcaa cattggcggc
gtagtcgtca agaaccaagc cgttgagctg 480gcagagaaga tgtccagcac attcgcccaa
ggcgaagggg acggattgct cggtctagca 540ttcagcaaca tcaacacggt acagccaaag
tccgtgaaaa cgcccgtcga gaacatgatc 600ctgcaggatg acattcccaa gtcggctgag
ctgttcacgg ccaagctgga tacctggcgg 660gacactgatg acgagtcgtt ttacaccttt
ggcttcattg accaggatct ggtgaagacg 720gcaggtgaag aggtctacta cacccctgtc
gataacagtc aaggcttctg gctattcaac 780tcgacctccg cgacggtaaa tggaaagacc
attaaccggt cgggtaacac cgccattgct 840gataccggta cgacgctggc cttggtggac
gatgacacgt gtgaggccat ttatagtgca 900attgacggcg cctattatga tcaggaagta
cagggctgga tctatccgac cgatacggcg 960caggataagc tacccactgt gtcgtttgcc
gtgggtgaaa agcagttcgt ggtgcagaag 1020gaggacctgg cgttttcgga ggcgaagacg
ggctatgtct atggaggaat ccaaagtcgt 1080ggtgatatga ccatggacat cttgggagac
acatttttga agagtattta tgctgtaagt 1140gcattgctgt tggcgttaag gggtgatatc
gaagctcact aa 118274849DNAAspergillus niger
74atgaagttct ctaccatcct taccggctcc ctcttcgcca ctgccgctct ggctgctcct
60ctcactgaga agcgccgtgc tcgcaaggag gcccgcgccg ctggcaagcg ccacagcaac
120cctccctaca tccctggttc cgacaaggag atcctcaagc tgaacggcac ctccaacgag
180gattacagct ccaactgggc tggtgccgtc ctgatcggcg acggctacac caaggtcact
240ggcgagttca ctgtccccag tgtctctgct ggatctagca gctccagtgg ctacggcggt
300ggctacggct actacaagaa caagagacaa tccgaggagt actgcgcctc cgcttgggtt
360ggtatcgacg gtgacacctg cgagaccgct attctccaga ctggtgtcga cttctgctac
420gaggatggcc agacttccta cgatgcctgg tacgagtggt accccgacta cgcctacgac
480ttcaacgaca tcaccatctc cgagggtgac accatcaagg tcactgtcga ggccaccagc
540aagagcagcg gtagcgccac cgttgagaac ctgaccactg gccagtccgt cacccacacc
600ttcagcggca acgtcgaggg tgacctttgc gagaccaacg ccgagtggat cgtcgaggac
660ttcgagtctg gtgactctct tgtggctttc gctgacttcg gctccgttac cttcaccaat
720gctgaggcta ccagcgacgg ttccactgtc ggcccctctg acgctaccgt tatggacatt
780gagcaggatg gcaccgtcct caccgagacc tccgtctctg gcgacagcgt cactgtcacc
840tacgtttaa
84975822DNAAspergillus niger 75atgggagatt acggccccgg agtgtcgtca
ctcacggcac agctacctgg aaatccgcct 60gtctctgaaa cagatcagga tgagatctca
gtacttgtaa cgggctttgg gccattcaag 120tctaatctag tgaacgcctc atatctgata
gcctcgtccc taccaccctc tttcacattc 180tcacctgcat cttcagacgg ctctgatgct
gttccccgtc gagtttcgat aaatgtccat 240ccttcaccca tacccgttgc atattcatcg
gtgcggacga ccctccccgt cattctcgat 300gactatgcca agacgcacgg aggccgacgc
ccagacatcg tcatacacat tggcatagca 360gcaatgagga actactattc cgtggagacg
caggctcacc gtgatgggta tctgatgtcc 420gacatcaaag gcagatccgg gtacgaggat
ggcgagaagc tgtggaggga gctcgacttg 480ccactggtgc ttagggctgg cccttcagag
ggacacgcct cggagaagaa acatctcagc 540ccccgtccac cggacgaaga tttcctagca
gcatggaaga cattttgccc tccagaaacc 600gatgcgcgga tctccactga tgccggacgt
tatctctgcg agttcatcct gtacaccagc 660ttggcactgg cataccaggc gggtgaggat
cgcaatgtca ccttcttcca tgttcccgcg 720tcatgcttgg atgaggatat agagacgggc
aaggaggttg ccgtcgcgct aatcaaggct 780cttgtgacta gctggagtga gcagcagcac
agcgttccct ag 822761629DNAAspergillus niger
76atgggctcaa ggcagggaaa ggcccccttt ggctggggta ctcagtcact tgctcacttt
60ggtatcaacc cagaccttgg gttgcacaac cagcagaacc tcaactccct catttcacat
120tcagcgatgg ccactgcgtt ggagacggaa tatgccacca tccctattga ccataacaac
180gcatcggctg gcacttatca aaatcggttc tgggtcagcg atgaattcta tcagcctggc
240aacccgatat ttgtgtacga taccggggag tcggatggcg gatcgatagc ccagtcctac
300ctaacctcca ctctctcctt cttcagagaa ttcctgatcg aattcaacgc catgggaatc
360gcctgggagc acagatacta tggaaactcg accccggctc ccgtatccta tgaaactcca
420cccgaggcat ggcaatacct caccaccaag caggcgctcg cggaccttcc gtactttgct
480agtaacttta gccgcgagaa gtatcctgac atggacctga cgccgcaggg cacgccgtgg
540atcatggtgg gcggctcgta cgcagggatt cgtgctgcat taactcgcaa ggagtaccca
600gagacgatat tcgcagcctt ttcctcatcg tctccggtgg aagcacaggt caatatgagc
660gcgtattacg accaagtcta tcgtggcatg gttgccagcg gatggaccaa ctgctcggca
720gatatccacg ctgctctgga atatattgac gatcaacttt cggatgaaga tacagctacc
780tcggtcaaac aacttttctt cggatctggc gccgagacca actccaacgg tgatttcact
840gcagcgctaa ctgccatcta cggctacttc caaagttatg gtatggcggg aggtattgga
900ggtctaggcg cattctgcga gtatctcgaa attgatccca agacgaacgg gactacagga
960ccggatggcc ttgcccctac gtatggcggc cagtatgtcg ccgaacgatg ggccgcatgg
1020ccaacctttc tcgagctggt caatctgaat atggggacca actgcgggcc tcaggacgcg
1080tctcagccaa ttgactgtga cttttccaag ccatacggcg atccctcggc catcacttgg
1140acttggcaat actgcagcga atgggggttc ttccaggcga acaacgatgg gccgcactcg
1200ctggcctcgc gatatcagtc ggtggaatac cagcaagaag tatgtaaccg gcagttcccc
1260gatgcagtgg acaagggact gctgcctccg tcgccgcggg cggatgatgt caaccaagag
1320tttgggggat ggacgatccg cccgtccaat gtttacttca gcggaggaga attcgatccg
1380tggcgatcat tgtccattct gtcgacagaa gatttcgcac ctcaaggggt ggagtttacg
1440agcgcgatcc cagcctgtgg ggtgcagacc aatgaggaca ccgtctttgg atacgtcatg
1500cagaactcgg aacattgctt tgactttcaa gcgacgccga ccgtggggaa gttatcacgc
1560ggcatcttca catccgcctt gttgcaatgg ctcgaatgtt ttggacagaa ctcaagccaa
1620tccaggtga
1629771176DNAAspergillus niger 77atgaagctct caatagctct tgcactcggc
gcaacggctt cgacgggggt gttggctgct 60gttgtaccgc agcaagaacc gctgataacc
ccccaagatc ccccaactca tcatcatcag 120gagaagttct tgatcgagtt ggctccttat
cagacgagat gggttaccga ggaagaaaag 180tgggacttaa aactggatgg cgtgaacttc
atcgatatta ctgaagaacg aaacactggg 240ttctacccaa cgttgcatgc tggtagctat
gttcactatc cgccgacgat gaagcatgcg 300gagaaggtgg ttccccttct gcggggtctc
tccaaggaca acatggagca aaacctcaac 360aaatttacct catttcacac tcgctactat
aggtcgtcca ctggtattga gtccgcaaag 420tggctataca gtagggtttc ggatgtcatt
gagcagtcgg gtgcagcaga gtacggcgcc 480actgtggagc agttcgctca ctcatggggc
caattcagta tcattgctcg gatcccaggc 540cagactaaca aaactgttgt cctgggcgca
catcaggaca gcatcaatct tttcctcccc 600tccatcctag ctgcacctgg tgccgatgat
gacggaagtg gaaccgtgac tatactcgaa 660gctttgcgtg gtctgctgca gtcagacgcc
attgtccggg gcaacgcttc caacacaatc 720gaattccact ggtactcggc agaggaaggt
ggtatgcttg gttcgcaagc catattctct 780caatataaga gagataagcg agacatcaag
gcgatgcttc aacaggatat gactggttat 840acccagggag ctctggacgc cggtcgtcaa
gaagccattg ggattatggt tgactacgtt 900gatgagggac tgacacaatt cctcaaagat
gtcactactg agtattgtgg tattggctac 960atcgaaacca gatgtggcta cgcctgttcg
gaccacacgt ccgcaagcaa atatggctat 1020cccgcagcta tggcgacgga atccgaaatg
gaaaacagca acaagaggat ccacacgact 1080gatgacagca tccggtatct aagcttcgat
catatgctgg agcatgcgag gttgacactt 1140ggcttcgctt acgagctggc ctttgctcaa
ttctag 1176781329DNAAspergillus niger
78atgagaacta ctacgtcttt tgctaggctt gcattggcag tggcctcagt tggtattgtc
60tttgctagtc caacaaaaaa taacgatggg aaactggtat atggctcacc agaatccgtc
120ggcatgatat ccgccccttt gcaccaaatg gtccaaaatg ttagcgcata tacacatgct
180gccaactata gcaagttctc gtacgacaaa gtccatccca tcgagccagg gtctgttacc
240ctggtggctc tcgacggtgt catcgtcagc gaatttgcct tgggcaagag aaatctctac
300gccgatgtca acggcaccaa tttacctcga tacctgcagg aagacaccac cctggataca
360gtctacgata tggcaagcct cacgaagctg ttcaccacgg tagctgcttt acgggaactt
420gacgctggtc gaattgcgct taatgtaact gttgcaactt atataccgga ctttgcgacg
480aatgggaagg agaatattac tatcttggag ctgttcacgc atacaagcgg tttcgcttct
540gatccatcgc caccactttt ctctgcttat tatacgacgt atgatgaacg cattaaagca
600attttgacgc aaaaaattat caataccccc ggcagcacat acctctactt agatctcaac
660tttatgtcgc tgggcctcgt tatcgagacc gtaacgggac gtgccctgga tgatcttatt
720tatgacttca ccagaccgct tgaaatgaca tctaccttct tcaaccgcgg gaatatcgaa
780ggctctacac cccagtcacc caactacgac cgcacagccg tacaagaatt tcagatcgca
840gccctcggac cctcagaacc acagcgtcca caaccagtgc gcggcacagt tcacgacgag
900aacgcatggt ccctagacgg cgtatcaggt catgcaggtc tattctccac tgtgcgcgat
960acagcgacat tctgccagat gatcctcaac aacggcacat atgcaggcca acggatcctt
1020tctcgaacag cggtagacat gattttcaca aacttcaatg ccaggtttcc gggggatgct
1080cgtagtttag ggtttgagtt ggatcagtat tctactgcgg gaccgatggc gagtttgcaa
1140actgcgagtc acactggatt tactgggact acgttggtga tggataggac gtataacgcc
1200ttttggttgc attttagtaa ccgggtgcat ccgtctaggg catggtctag caatactatt
1260gtgagagagg ctattgggta ttgggttggg aagagcttgg ggttggatgt tgcgtttgct
1320ctgttgtaa
1329791839DNAAspergillus niger 79atggcgtcct ggttgctctc gacgctcctt
tttctgagcc cgtccttggt gtcagccaaa 60tcggccgcag actattatgt tcactccttg
cccggtgccc ccgaggggcc cttgctgaag 120atgcatgccg gccatattga ggtggatcca
cagaacaatg gaaatctttt cttctggcac 180taccagaatc gccatattgc caaccgccag
cggactgtga tctggttgaa cggtggtccc 240ggatgtagtt ccatggacgg cgcgttgatg
gaggtcggtc cgtatcgcct gaaggacaat 300gaaaccttga cctataatga gggttcctgg
gacgaattcg ccaatttgtt gttcgtcgat 360cagccagtcg gaaccgggtt cagttatgtc
aacacggaca gctatcttca tgagctcgat 420gagatgtcgg ctcagttcat tgtctttctg
gaagagtggt tcagattatt tccggagtat 480gaacgcgatg atatctacat tgccggcgag
tcttacgccg gtcagcatat tccatacatc 540gccaaagcca tccaggaacg gaacaagaac
gttcaaggga agaccatcgc ttcgtggaat 600ctaaaaggcc tattgattgg caatggttgg
atttctccta atgaacagta catgtcctac 660ttgccctacg catatgaaga aggccttatc
aaggaaggca gccggaccgc gaaggaactc 720gaagttttac agtcagtctg taagtccagg
ctggaaactg gcaagaacaa ggtccacctc 780aacgactgcg agaaggtcat gaatgctctg
ttggataaga cggtcgaaga caacaaatgt 840ctcaacatgt atgacatccg ccttcgtgac
accaccgatg catgcggtat gaactggccc 900accgacctgg aggacgtgaa gccctatctg
cagcgggaag atgtggttaa agcgcttaac 960atcaatccgg agaagaagtc tggctgggtg
gagtgttcag gtgcagtgag cagcgctttc 1020aatccgcaaa agtccccgcc ctcggttcaa
ctacttcccg gcttgctgga atcgggactt 1080caaatcctcc ttttcagcgg agacaaggac
ctgatttgca accatgttgg aacggaacag 1140ctcatcaata acatgaagtg gaacggaggc
acgggtttcg agacctcacc tggcgtctgg 1200gctcctcgac acgactggag tttcgaaggc
gagccggcgg gtatctatca atatgccaga 1260aacctgactt acgtgctcat ctacaacgca
agccatatgg ttccctacga ccttcctcgt 1320cagagccggg acatgctaga tcgcttcatg
aatgtcgata tcgcgagcat cggaggcagc 1380cccgccgact cgcgcattga cggcgagaag
ctgccccaga cgtcggtggg cggccatccc 1440aacagcaccg cggcggagga gcaggagaag
gagaggatca aggagacgga atggaaagcc 1500tacgccaagt caggcgaagc cgttctcctc
gtcgtcatta tcggtgtatt agtttggggc 1560ttcttcatct ggcgcagccg ccggcgtcac
cagggatacc ggggcgtctg gcataaggac 1620atgagcggaa gctctgttct cgagcggttc
cacaacaagc gcacgggagg cgcagacgtc 1680gaagcggggg atttcgacga ggcggagctc
gatgaccttc attctccaga cctcgaaaga 1740gaacactacg ccgtgggcga ggacagcgac
gaggatgata tttcacgaca gcattctcaa 1800caggcctccc gagccggggg cagtcataat
ctatcctag 1839801596DNAAspergillus niger
80atgtttctga tttcacctgc agtgacagtt gcggctgcac ttctgctgat caacggcgca
60ggagcaactc aatctgaacg aagtcgggct gccgctcatt tttccaaacg tcatccgacg
120taccgtgctg cgaccagagc ccagtcgagc aacacttccg actaccgatt cttcaataat
180aggaccaagc cccacttggt ggaaagctta cccgatgtgc acttcgatgt tggggagatg
240tactcggggt cgatccctat cgatgacagc aacaatggat ctcgatccct gttttatatc
300ttccaaccta agataggcga accttcagac gaccttacca tttacctcaa tggagggcca
360ggctgttcct ccgaacaggg attctttcag gaaaatggca ggttcacatg gcagcctggt
420acctatgcac ccgtcatcaa cgaatattct tgggtcaatt tgacgaacat gctatgggtt
480gaccaaccag tcggaaccgg attttccgtt ggaaatgtta cagccaccaa cgaagaagag
540attgccgccg attttctcga cttctttgaa aagtttgaag atctatacgg gataaagaac
600tttcgcattt tcatgaccgg tgagagctac gccggtcgct atgttcccta tatctcgtcg
660gcaatgctag acaagaacga caccacgcgt ttcaatctga gcggagccct tctttatgac
720gcctgcatcg gccaatggga ctacatccag gccgaactcc ctgcctaccc cttcgtcaag
780cagcacgctt cactattcaa cttcaatcag tcctacatga acgagcttga aaccacctac
840gaagaatgcg gctacaaggc ctacttcgat gagtactttg cctttccacc aagcggcatc
900caacccccaa aatacatgaa ctactccgag tgcgacatct ataacatgat ctactacgaa
960gcctataacc cgaacccatg cttcaatccc taccgcgtca ttgatgagtg tccacttctc
1020tgggacgtcc tgggctggcc gacagacttg gcatacgagc ctgcgcccac cacatacttc
1080aaccgtatcg atgtcaagaa ggccctgcac gcccccatgg atgtggaatg ggagctctgc
1140agctacgacc tcgtcttcgc tggaggcgac gctgacccgg gtccggagca gcaaggggat
1200gactcaccca accccaccga gggtgtcctc ccgcgtgtta ttgaggcgac caaccgcgtg
1260ctcattgcca acggtgactg ggactacctg attatcacca acggcaccct cctcgccatc
1320cagaatatga cctggaacgg ccagctgggc ttccagtccg cacctgccac accgatcgat
1380attcagatgc ccgatctcca gtgggttgag atttttgagg cccaggaggg atatggaggg
1440ctggatggcc ctcagggggt tatgggtgta caacattatg agcgcggttt gatgtgggcg
1500gagacatatc agtcggggca taagcaggct caggatcagg gccgtgtctc gtatcgccat
1560ctgcagtggc tgttggggca agttgagatt ctttag
1596811596DNAAspergillus niger 81atgctgtttc gcagtctgtt gtcgacggct
gtcctagccg tctcgctgtg cacggataat 60gcttcagctg ctaaacatgg tcgatttggc
caaaaagctc gcgacgccat gaacatcgcg 120aagcgttccg ctaacgccgt gaaacactcg
ttgaagatcc ctgtcgagga ctatcagttc 180ttgaacaaca agactaagcc ttaccgcgtg
gaaagcctgc ctgatgttca cttcgatctg 240ggcgagatgt attccggctt ggtccctatt
gagaagggca acgtgtcacg gtcccttttc 300tttgtcttcc agcccactat tggcgagcct
gtggatgaga tcaccatctg gctgaatggt 360ggccctggtt gcagttccct tgaggccttt
ctccaggaga atggtagatt cgtgtggcag 420cctggaacct accagcctgt tgagaaccca
tactcgtggg tgaatctcac caatgttctg 480tgggttgacc aacctgtggg aacgggattc
tctctgggtg tcccaaccgc tacgtccgag 540gaggagattg ctgaagactt tgtgaagttc
ttcaagaact ggcagcagat ctttgggatc 600aaaaacttca agatctatgt tactggagaa
agttatgcgg gccgttatgt tccttacata 660tccgctgctt tcctagatca gaatgataca
gaacacttca acctaaaagg tgcactggca 720tatgatccct gtattggtca gtttgactac
gtgcaggagg aagcacctgt tgttcccttt 780gtccagaaga acaatgccct cttcaatttc
aatgcaagct ttttggcgga actagagagc 840atccatgagc aatgtggata caaggatttc
atcgaccagt atctagtctt cccagcatcc 900ggtgtccagc cgccaaaggc tatgaactgg
agcgatccca cctgtgatgt ttatgacatc 960gttaataacg ccgtcctgga tcccaacccg
tgcttcaacc cctacgaaat caacgagatg 1020tgccccattc tctgggacgt tcttggattc
cccaccgaag tcgactatct ccctgcgggc 1080gccagcatct actttgaccg cgctgatgtt
aagcgtgcca tgcacgctcc taacatcacc 1140tggtccgagt gctcggtgga gagcgtcttt
gtcgggggcg acggcggtcc cgagcaggag 1200ggcgactact cggccaaccc catcgagcat
gtcttgcccc aggtcatcga aggcaccaac 1260cgagttctga tcggtaacgg tgattatgac
atggtcatcc ttaccaacgg cacccttctc 1320tcgatccaga acatgacatg gaatggaaag
cttggattcg acacggcccc cagcaccccc 1380atcaacatcg acatccctga cctgatgtac
aatgaagtgt tcattgagaa cggctatgac 1440ccacaaggtg gtcagggtgt catgggcatc
cagcactatg agcgtggtct tatgtgggct 1500gagaccttcc agagcggaca catgcagccc
caattccaac ccagagtgtc ataccgtcac 1560cttgagtggc tgcttggccg gcgggatacc
ctgtaa 1596821479DNAAspergillus niger
82atgaaaggtg cggcgctaat tcctcttgcg gcgggcattc cttttgccca tggcctgtct
60ctccataaac gcgacgggcc tgccgtcgtt cgtatgccca ttgagcgcag gagcgcccag
120tccttgcaga aacgagattc tacggtcggt gtgactttgc agaactggga tgcgacctat
180tacgcagtca acctgacgtt aggaacacct gcgcaaaagg tatcattagc tttggacact
240ggcagcagcg acctctgggt gaacaccggc aactcaactt actgctcaat cgacaatcta
300tgcacccctt atggcttgta caatgccagc gaatcgtcta ctgtaaagac cgtgggcaca
360cacctcaacg atacatatgc ggacggcaca aacctttacg gtccttatgt gaccgataag
420ctcacgatcg gcaacacaac aatcgataat atgcagtttg ggatcgccga gtcaacgact
480agtaaacgcg ggatcgccgg cgtcggttac aagatttcga cctaccaagc cgagcatgac
540gacaaagtct acgccaacct ccctcaggcc ctcgtcgaca gcggtgccat taagtctgct
600gcgtacagca tatggctaga tagtttggag gcgtcgactg gctccctcct tttcggaggt
660gtcaatacag ccaagtacaa gggcgatctg cagactcttc cgatcattcc tgtgtatggc
720aaatactact ccctcgccat cgcccttacg gagctcagcg ttgcgaccga ctccaactcc
780agtagcttca ccgacagtct ccccctctct gtgtcactcg atactggcac caccatgacg
840gcactgccca gcgacctggt caacaaggtc tacgatgcgc tcaacgcaac ctacgacaag
900acatacgaca tggcctacat cgactgcgac actagagagg cggattacaa tgtaacatac
960agtttctccg gggcaacgat caccgtgagc atgagtgagc tgattatccc cgcaacggag
1020ccggggtggc ccgacaacac gtgtgtcttg ggcctcgtgc ctagccagcc gggcgtgaac
1080ctgctcggtg atacattcct gcgcagtgcg tacgtcgtgt atgatctcga gaacaacgaa
1140atctctctcg ccaataccaa tttcaatcca ggcgacgatg atatcctcga aatcggaacg
1200ggaacgtctg ctgtgccagg agccacaccg gttccctctg ctgtctcttc tgcaactgga
1260aatggactga tctcgtctgg caccgcagtg cccacgctgt cgggtgtcac aataactgct
1320acagccacag caaccggctc aaccggcact ggctctagcg gtggttcgtc ggctgaagcc
1380acgagtactt cctcggaggg cgctgcggcg caagctacga gcaacccgat gaacctgctc
1440ccaggacttg cgggtatcgg cctacttctc gctctgtaa
1479831836DNAAspergillus niger 83atgctgtcgt ctctccttag ccagggagca
gccgtatccc tcgcggtgtt gtcgctgctc 60ccttcgcctg tagccgcgga gatcttcgaa
aagctatccg gcgtccccaa tggctggaga 120tacgccaaca atcctcaagg caacgaggtc
attcgcttgc aaatcgccct tcagcagcat 180gatgtcgctg gtttcgaaca agccgtgatg
gatatgtcca cccccggaca cgccgactat 240ggaaagcatt tccgcaccca cgatgagatg
aagcgcatgt tgctccccag cgagactgcc 300gtcgactcag tccgcgactg gctggaatcc
gccggtgtcc acaatatcca ggtcgacgcc 360gactgggtca agttccatac caccgtaaac
aaggccaatg ccctgctgga tgccgacttc 420aagtggtatg tcagcgacgc caagcatatt
cgtcgtctgc gcaccctgca atactccatc 480cccgacgccc tggtctcgca catcaacatg
atccagccca ccacccgctt tggccagatc 540cagcccaacc gtgccaccat gcgcagcaag
cccaagcacg ccgatgagac attcctcacc 600gcagccaccc tggcccagaa cacctcccac
tgcgactcca tcatcacacc gcactgtctg 660aagcagctgt acaacatcgg tgactaccag
gccgatccca agtccggcag caagatcggc 720tttgccagct accttgagga atacgcccgg
tatgccgatc tcgagaggtt cgagcagcac 780ctggctccca atgccatcgg ccagaacttc
agcgtcgtcc aattcaacgg cggcctcaac 840gatcagcttt catcgagtga cagcggcgaa
gccaacctcg acctgcagta catcctgggc 900gtcagcgctc ccgtccccat caccgagtac
agcaccggcg gacgcggcga actagtcccc 960gacctgagct cccccgaccc caacgacaac
agcaacgagc cctaccttga cttccttcag 1020ggaatcctca agcttaacaa ctccgacctc
ccacaagtca tctctacctc ctacggtgaa 1080gacgaacaga ctatccccgt cccctacgcc
cgcaccgtct gcaacctcta cgcccaactc 1140ggcagccgcg gcgtctctgt aatcttctcc
agcggcgact ccggcgtcgg cgccgcctgc 1200ctcaccaacg acggcaccaa ccgcacgcac
ttccctcctc aattccccgc ctcctgcccc 1260tgggtaacct ccgtcggcgc aacctccaag
acctcccccg agcaagccgt ctccttctcc 1320tccggcggct tctccgacct ctggccccgc
ccctcctacc aacacgccgc cgtgcaaacc 1380tacctcacca agcacctggg caacaagttc
tcggggcttt tcaacgcctc cggccgcgcc 1440ttccccgacg tctccgcgca gggcgtcaac
tacgctgttt acgacaaggg catgcttggc 1500cagttcgacg ggacgagttg ctccgcgccg
acgttcagtg gcgtcatcgc gttgttgaac 1560gatgcgagac tgagggccgg gttgcctgtg
atggggttct tgaatccgtt cctgtatggt 1620gtcggaagtg agaagggtgc gttgaatgat
attgtgaacg gcgggagtgt gggttgtgat 1680gggaggaatc ggttcggggg cacgcctaat
ggtagtcctg ttgtgccgtt tgctagttgg 1740aatgccacga ccgggtggga tcctgtgtcg
gggttgggaa cgccggattt tgcgaagttg 1800aaaggggtgg cgttgggtga ggagggtggt
aattaa 1836841437DNAAspergillus niger
84atgtggctct ttctcgtgtg cagtatcctg ctgccacttg gagtagtcaa cgcacagtct
60caatacttca acaacaaaac caaagaattc gtcgtcaatg gctctgctat tccttttgtc
120gatttcgaca ttggcgagtc ctatgcgggc tacctaccca acacgccttc tggaatctcg
180agtctatact tctggttctt tccatcttct gatcctgatg cgtctgatga gatcaccgtc
240tggctgaatg gcggcccagg atgcagctct ctggcaggca tcatgctcga gaacggcccc
300tttctatggc aacctggtac ctaccgaccc gtgcgcaacc cttatgcctg gaacaacctc
360acaaatatgg tgtacattga tcagcctgct ggaacgggat tctcgcttgg cccgtctacg
420gtggtctcag aatttgatgt agccagacag tttatggact tctggaggcg gttcatgaaa
480acattcgatc tgcagaatcg aaagatatat ctcactggcg agagctatgc gggccagtac
540atcccataca tcgcgtcgca gatgcttgac caggatgatg atgagtattt ccgggttgcc
600ggcatccaga tcaatgatcc ctacatcaat gagctgccag ttttgcaaga tgttgcgacc
660gtcaatcagc accgctccct ctttcccttt aatgacacct tcatgagtca aatcaccaag
720ctttccgacg attgtggcta cacttcgttt cttgacgatg cccttacctt tccaccccgt
780tctcaattcc catcagtgcc ctataatgct agctgcaaca tctgggatat cataaacaac
840gcttctctag ctctcaaccc atgcttcaac cgctaccata tccccgacgc ctgccccacc
900ccctggaacc cagtcggcgg ccccatcgtt ggacttggtc cgaccaacta cttcaaccgc
960agtgacgtcc agaaagccat caacgcgtac ccaacggact atttcgtctg caaggatgga
1020atcttcccga cggccaacgg actggacaca tcccctccaa gctccctggg accgctgccg
1080cgcgtcatcg aacagaccaa caataccatc attgcgcacg gcctgatgga tttcgagctg
1140ctggcgcagg gaaccctgat cagtatccag aatatgacct ggaatgggaa gcaggggttc
1200gagcgggagc cggtggagcc gttgttcgtg ccgtatggtg gatcatcggg aggaggcgtg
1260ctgggaacgg cacatacaga gcgtggattg acattttcga cagtatttag ttcaggacat
1320gaaatcccgg aatatgcacc gggggcggca tatcgccagc tggagttttt gctggggagg
1380gttgcgaatc tgtcgacaat tattgagcag gtgcagataa cagagcagaa tggttga
143785633DNAAspergillus niger 85atgtccaaac tctccgctgc tatctccaag
ctctccctct ccaccatagc caccactctg 60ctcctcctta cccccccaac caccgcctac
ttctacaaat atcccgccct cttcgtctac 120aaagacacca actgcaccga tatctccttc
tcacttgtct acccctccct gggtaactgc 180aacggcggat actacgacta cgcgggctca
ttccagatgt tcaatatcga tgctgcgtat 240acctgtaatg gcagtgactc gacactgatg
tttgagatgt ataatagctc cggctcggat 300tgtggagatg agagtgattt gttgtttaga
cagccggtga cggaggagtg tactgttgcg 360gatgtggaga gtccggggcc gttggagatg
ccggtttggt ttgagttggg gtcactattg 420gggaattgtg gtgggatggc tggtactatg
ttgttcggtg tggggattct tgagggtggg 480ttagagacta aattatactg gaaatgttat
tcatcaaggc tgaatacaag tgtaaccgtg 540cacagattat ctttgatact gtctatgggc
tgtacgagcg tctctgactc ctacaatgag 600ttagcggctg cacattacta tgaggacctg
tga 633861827DNAAspergillus niger
86atgcgtcacc tcttatcact gctggtgctt ctgatcgcat cggccgccct ggtctccgcc
60gtccccgccg gctccattat cactccacaa ccacccgtcg agcccgttca ccttctctct
120tcccagccct ctgatccccg aaggccatgg atccgcctcc gtgactggat catcgagtcc
180atatggggca tcgaaaaacc cgcatctcgt cgattcccac tcaacgattc cccgcgcaat
240cgctctcctc cctcccggat tctggcgcgc tacggtagtg acgtcgtact tcgtttcagc
300ctgcgcaatc acgatgaggc cgaggcattg gcccaggctg cagacattct attcctggac
360gtatgggcgt ctactccagc attcgtagat atccgactgg ccgaggaagt caccgcatat
420actcccctaa tagacaacct ggcagagaga atctatacga cctatccatc taaaaagccg
480ataggacttg aaggacaatc tggatttgcg tcctcgagtc gacctgcgcc aaagttcggt
540gacctttttt tccacgagta tcagcctttg tccgtcatta tcccctggat gcggctgctg
600gcttccatgt ttccatccca tgtgcgcatg attagcgttg gagtatctta cgagggtcgc
660gaaattcccg ccctccgact gagcgcaggc agctccaccg cggcgtcagg ccctcgtaaa
720acaatcatcg ttacgggtgg tagccatgcc cgcgaatgga ttggcacctc aaccgtgaac
780catgtaatgt acacgctcat taccaagtat ggcaaatcca aggccgttac ccgccttcta
840caggacttcg actggatcat gatccccacg atcaatcccg acggctatgt ttatacctgg
900gagacggacc gactatggcg caagaatcga cagcggacca gcctacgctt ctgtcccgga
960atcgatcttg accgcgcctg gggcttcgaa tgggacggcg gtcggacccg cgctaaccct
1020tgttcagaaa actatgctgg agacgagccc ttcgagggaa tggaagcaca acaattagca
1080cagtgggcgc tcaacgagac acaaaacaac aatgccgaca tcgtgagctt ccttgacctt
1140cactcttact ctcaaacaat tctctacccc ttctcctact cctgctcctc gatccctcca
1200acgctcgaga gcctggaaga gctaggcctt ggcctagcca aggccattcg gtacgcgact
1260cacgaaatct acgatgtcac ttctgcctgc gaaggcatcg tcacggccag tgcggcagat
1320aacaaccccg ggcggttctt ccccattggt ggcaactccg gtggcagtgc gttggactgg
1380ttttaccacc aagtgcacgc gacttattca taccagatca agcttcgtga tcgcggaagc
1440tacgggttcc tccttccgtc tgaacacatc atccccaccg gcaaggagat ctacaatgtt
1500gttctgaaat tgggatcctt cctcatcgga ggcgactcat ttgacgtcga ttgggaatca
1560gaactcttcg atctgtcaaa ggacgaatcc gatctggata gccgctattc aaaatccaat
1620gaccgctccc cggcgtatct acacaacgcc aacggccccc tgcccaacat tgacgaagac
1680gaagataagg aatgggtaat ggtggaggaa gaagactaca cagacgatga cgacgacgat
1740gatgatgatg atgaagaaga ggaagaggaa gaggaagata catattgggc caccgaacac
1800acatacgaat ttcggcgacg acgctga
1827871251DNAAspergillus niger 87atggctttcc tcaaacgcat tctcccgctg
ctggccctca tcttacctgc agttttcagt 60gccacagaac aggtccctca tccgaccatc
cagaccatcc cggggaagta cattgttact 120ttcaagtccg gcattgacaa tgcgaagatt
gagtctcatg ccgcatgggt aacggagctc 180cacaggcgca gcttagaagg ccgcagtaca
accgaagatg accttcccgc cgggatcgaa 240agaacgtaca gaattgccaa ttttgctggg
tacgcggggt ctttcgatga gaaaactatc 300gaggagatcc gcaaacatga ccatgtagct
tatgtggaac aagatcaggt ctggtatctc 360gatacgctag ttaccgaaag gcgagctcct
tggggactgg ggagcatttc tcaccgtggt 420gggtctagca ccgactacat ctatgatgac
agcgctgggg agggtacata cgcttatgta 480gtggacaccg gcatcttggc tacgcataat
gagtttggtg gtcgtgctag cctggcatat 540aatgctgcag ggggtgagca cgttgatgat
gttggacatg gtacacatgt agcagggacc 600atcgggggca aaacatacgg ggtttcgaaa
aacgctcacc tactgtccgt gaaggtgttt 660gtaggtgaat ccagctcgac atcggtcatt
ctggatggct tcaattgggc cgccaatgat 720attgtgagca agaaccggac cagtaaggcg
gcgataaata tgagtcttgg tggaggctac 780tcctatgcgt ttaacaatgc agttgagaat
gcttttgacg agggtgtgct ctcttgtgtt 840gccgctggaa atgagaatag agatgcagca
cggactagcc cggcttctgc acccgacgcc 900attactgttg ccgctatcaa cagaagcaat
gcccgtgcgt cattctcaaa ctacggctct 960gtggttgaca tttttgcccc gggagagcaa
gtactttctg catggaccgg ctcgaactcg 1020gccaccaaca cgatctccgg cacgtccatg
gctacacccc atgtgacagg tttgatcctc 1080tatttgatgg gcttgcggga ccttgctacc
ccagcggctg caacgaccga gctcaagagg 1140ttggctacgc ggaatgctgt caccaatgtg
gcgggtagcc ccaatcttct ggcctacaat 1200ggaaacagcg gcgtgtcaaa agggggtagc
gatgatggag atgaggacta g 1251881368DNAAspergillus niger
88atgatcaccc ttttgtcggc cctgttcggc agcgtagtat atgccgctac gcagaccgtg
60ttagggccag agggggctga tccctttacg gtgtttcgca gcccacactc accggcattt
120tcaattcgca tccaggagca gaatgactcg atctgtgatg ctcgttcacc ccaattcact
180ggttggctcg acattggccc gaagcatctt ttcttttggt attttgaaag ccagaatgac
240cccttccatg atcccctaac gctatggatg actgggggcc caggagactc gagtatgatt
300ggacttttcg aagaagttgg cccttgccgg attaatgagt ttgggaatgg aacagatcac
360aacccctggg cctggaccaa gaattcatca cttctttttg ttgaccagcc agtcgatgtc
420gggttttcct atatcgatga gggctatgag ctgcctcatg actcacgtga agccgcggtg
480gacatgcatc ggttcttgcg attattcata tccgagattt ttcctcacaa acagttcctt
540cccgttcacc tttccggtga atcttacgca ggccggtaca ttccttatct ggcgacccaa
600atcttggaac aaaatgaact gtataaagat agccccagga taccgctgaa atcgtgcttg
660gtgggtaacg gattcatgtc acccaaggat gcaacgttcg ggtattggga aacactgtgt
720actactaact caggagtccc atctcctatc ttcaatgaaa ctaggtgcga tattatggcg
780gcgaatatgc cgcactgtat ggatctatat gacatatgca ttcaacactc agaccccgcg
840atatgtcatg cggcccagtc cgtctgttac gatagtgttg tagggctcat ggccaaatta
900ttgctaagga tgacgacagt cactgcacct tgtgagatcg acgaaatgtg ctatatcgaa
960gcggctctaa ttgagagata tttgaattcg ccatctgttt gggaggccct gtcgccaccg
1020caacaggtta ccgaatacaa attcgtcgct acttctgtta ttgatgcatt tgctcaatca
1080gcggacggca tggtgtcgag ctcgaagcag atcgctttct tactcgcaaa taatgttgac
1140ttcttagcgt atcaaggcaa ccttgatctc gcctgtaata cggctggcaa cctacgttgg
1200gcgaactcgc tttcttggaa aggccagaca gaatttaccg caaagccctt acttccgtgg
1260gaaattcagg tttcggtcgg tgaagggacg gacgaaacgt cacgctttgc ctttgtgact
1320gtggacaacg ctggacacct gttgcgggac tcaaagattt caaactga
1368892376DNAAspergillus niger 89atgcggtttc tcacttattc cctgcccttc
attgcaagtg ctatctcgct ctttggggtc 60aatgtacaag ctcgatcaca agctccaagt
gccatccgtc atgtgtcgac gcttgaccag 120cccaccatca agacaccctc acagcgggtc
gatcaccttg accactttga catcaccttc 180aatattcatg acaagcacca gcggataaag
ctggagctgg agcccaacca tgacatcctg 240gcggaagacg catccgtaca gtatctcgac
gcggacggga acgtgcgacg gcacgagccc 300attgctccac atgagcataa ggtcttcaag
gggaggagtc tactcgggcg aggaaaaggc 360atgtgggatc cggtcggatg ggcgcggatc
tacttgaagc aggatggctc agagccacta 420tttgagggag tcttcagtat cgacggcgac
aaccatcacg ttcagctgaa atcggcatac 480atggagaaga aacgccccgt ggatgtcgac
cttcccgact cagcgactga ctatatgatc 540ttctaccggg attcggatat ggtgcgtcta
catacggaac tcaagcggtc gtcgctcgga 600tcgacctcgt gtcaagccga tcagctcggc
ttcaacacta accccaacca ccctgtgcta 660caaccgtatg gccaggcaga gaccgatacg
tggggagcaa tttcattgaa ctccttgttt 720ggactcaaca agcgccaatc cgatatcgga
agtgtgtctg gcaatgcggg cggagtcaat 780ctggcgtcga ccattggtga tacttcgggc
tgtccgagta cgaagcaagt agctttgatt 840ggtgttgcaa cggactgcgc ctttaccggc
tcattcaaca acgagactgc cgccaaggaa 900tgggtcatca gtactgtcaa cagcgcgtcc
aatgtctacg aaaagtcctt caacattacg 960attgggctgc ggaatctgac tatcaccgac
agctcatgcc ccgacaaccc gcccgcggcc 1020acggcatgga acatgccctg ctccagcggc
aatctcacct cccgactgga tctgttttcc 1080aagtggcgcg gtgagcaatc ggatgacaat
gcttattgga ccctgatgag cgattgcgcg 1140acgggcaacg aggtcggact gtcatggctt
ggccaactct gcaatagcga tgcttcttcg 1200gatggctcga gcacggtcag tggaactaac
gtcgtcgttc ggtcttccgg ctcggattgg 1260cagatctttg ctcatgaatc tggccacacc
tttggcgctg tccacgactg tgactcccag 1320acctgcgcgg aggatctcga agcctcgtcc
cagtgctgtc cgttgacctc gagcacctgc 1380aacgccaacg ggaaatacat catgaatcct
acaactggaa cagacatcac tgcgttctcg 1440caatgcacta tcggaaatat atgcgcagcc
ctgggccgca acagcgttaa gtccagttgt 1500ctctccgcca accgcgacgt caccacctac
actggcagcc agtgcggcaa cggaattgtc 1560gagtccggcg aagactgcga ttgtggcggg
gaagatggtt gcggcgacaa caactgctgc 1620gacgcgaaga catgcaagtt caagtcggga
gctgtgtgtg atgactccaa cgacagctgc 1680tgttcaagct gccaattctc ctcagctggg
acggtatgtc gtgccagtcg cggcgactgc 1740gacgtggcag agacctgcag cggcaactcc
agtacttgtc ctaccgactc gttcaagaag 1800gacggcacga gctgcggcag cagtggctcg
ggacttgcct gcgctagtgg ccaatgcacc 1860agccgcgact accagtgccg cagtgtgatg
ggcagtctcc tccacagcaa cgacacctac 1920gcctgttcct ccttcagttc ctcctgcgaa
ctggtctgca cctccccgaa gatcggcacg 1980tgctacagcg tcaaccaaaa cttcctcgac
ggcactccct gcggtagtgg cggctactgc 2040agcaacggcg actgcaaggg ccaaaacgtc
gaatcctgga tcaagaacca caaaggtatc 2100gtcattggtg tcgcctgcgc cgtaggcgcc
ctgatccttt tggccctgat gacctgcatc 2160gtaaaccgct gtcgccgggc tcgcgcgcca
aaacccgtcc cgcgtccagt gccttacggg 2220ccgtggcccg gcgctaggcc tcccccgccg
ccgcccatga accagtggcc ggcgcgaggc 2280tatcaaggct tagggaatga gccgccgccc
ccgtatccag gtgtacctgg tcagccagta 2340ccgcaacata tgcctcccca ggggcggtac
gcttga 2376901446DNAAspergillus niger
90atgcgtttcc taagcagtgc agccctattc ggcctggcgt atgcctccac ccaggcggtc
60ctccagccag aggaaccatc cgacttccgt acattccaca gcccatattc cccgcaccac
120tcgatccgca tccgccagca gaatgaatca atctgcgctg cccattccgc ccaatacacc
180ggctggctcg acatcggccg taaacatctc ttcttctggt actttgagag ccagaatgac
240cctgccaatg atcccctcac tctctggatg acaggagggc caggggggtc cagcatgatc
300ggtctgtttg aagaagtcgg gccatgtctg atcaatgagt acggcaatgg cacttactac
360aatccgtggg gctggtcccg gaactcctcc ctactatttg tcgatcagcc agtcgatgtg
420ggattttcgt acgtcgatga aggagaggac ctgccgggcg attcgcatca agctgcaatt
480gacatgcatc ggttcttgca gttgtttgtc tcggaggttt tcccgcaatt gcagactctt
540cccgttcatc tttctggtga atcgtatgct ggtcactatg tcccttacct cggcagtcag
600atcgtccaac agaacaagct ctatcccact gagccccagg tccttctgca ctcatgtctc
660gtaggcaacg gctactattc tcctcgcgac actacctacg gctactggga aaccctctgc
720accactaacc ctggagtccc cgagcccgtc ttcaaccgaa ccagatgcga catcatggcg
780gccaatatgc cgcgatgcat ggaagtatcc gacgtatgtg ttcggaaccc cgatccagct
840atctgccatg ctgcgtcgga ggtatgctac gagggcgtga tcggatggta tgatgacgag
900tctggtgaag gtggtcggaa taggtttgat ataaccgctc cctgcgccct tgacggcata
960tgctacatcg aggccgctcg catcgagcag tacctgaaca cacccgcagt ttgggctgct
1020ctatcaccac ccaaagaaat caaagaatac aaggttactt ccgacaatgt gtcgcgcgca
1080ttcgatctca cttcagacac gatgacgcca gcgtctgagc aagtcgcgtt cctgcttgcg
1140aatcaggtac atttcctggc gtatcagggc aatctcgatc tggcgtgtaa tacggcgggt
1200aatctgcgct gggcgcattc tctgccatgg agaggtcagg tcgagttcgc gtcgaaggcg
1260ctgcggccat ggagttgggt agatgtggta tctggaaaag gtggagtggc tggaacgacg
1320aaggaggagt cgaggtttgc gctagttacg gttgatgggg cgggacattt tcttcctcaa
1380gatagacctg atatcgcgtt ggatatgatg gtgcgctgga tatccggggc atcgtttact
1440gagtga
144691960DNAAspergillus niger 91atgacattgt tactcaactt ccacgcgctc
tttacagtca ttcttgttgc caatctttca 60accagatgca gcgcactgct ctctggacgt
gacttttgct ccacgccagc gcccggtgag 120tcactccgag cggagcatag gaggctgtat
gatgtacagg cccaacgtga cagcaccgcc 180gaggagagcc gggaggtggt gccttggatt
gaaatcgaga catggtttca tattgtaagc 240agcaatgaag cagcaaacac agtatcagac
gacatgatca ccagccagct ttcctatctt 300cagaaggcat atgaaagtgc gactatcacc
tatcggttgg agggcataac tcgtcacata 360aatgactcgt gggcgcgaaa tgatgatgaa
ctggggatga agaatgccct acgaaggggc 420aattatggca cattaaatgt ctatttccaa
acagatctcc aggcgtcatc cgacgagaat 480tctcgggact atccaaatga cggtaaccga
cgaacagatg tgtcagatca atcatcatca 540actgtcctag gcttctgtac gttgcctgac
ccgagtgtga attccagcag ccctcgttcc 600agctacatca aggatggttg taacgtgtta
gcggatatca tgccgggtgg tagtttagcg 660cagtacaaca aaggcggcac agcggttcat
gaggttggcc attggaatgg gctgctgcat 720acgttcgaag gtgaatcgtg ctcccctgat
aatgaaggag attacattga tgacaccccg 780gagcaatctg agcctacgag cggatgtccc
gccgagaaag attcatgccc cgatcttcct 840ggccttgatg ctattcataa ttttatggac
tattcatctg atgactgtta tgagagtttt 900actccagatc aagcggagag aatgaggagt
atgtggtccg ctatgcggga agggaagtga 960921920DNAAspergillus niger
92atgcatgtct cacttttcct actcagtgtt acggcagcgt ttgccagccc aacaccccat
60aactatgttg ttcatgagcg gcgcgatgca ttgcccagtg tctgggtaga agaaagccgg
120ctggacaaag gtgccctact gcctatgcgg atagggctta ctcagtccaa cctggatcgt
180ggccatgatt tattgatgga ggtgtctcat ccacaatcgt ctcgctacgg aaagcatctc
240tccagcgagg aggtgcacga cctatttgcc ccgtcgaatg aggccgtcga gaccgtccga
300acctggattg aatccgccgg aattgctcca agccgcatct cgcaatcata caacaagcag
360tggctacagt tcgatgccca tgcaagcgag gttgagcagc ttctgcagac ggaatactac
420atctacaccc atgccgacac gggaagttcc catgtgacat gccacgagta ccatgtgccc
480gaaaccatcc aatcgcacat cgactacata acaccaggag taaagatgct ggaagtgcgc
540ggcacgccct ccaaaaagag agatgcagag aagcgctctc ttggcagtct gcccccaatc
600ttagcaccac taccaatcaa tatcacgaag attttcgacg acccgctagc acactgcgat
660ctggcggtaa ccccagactg cattcgagcc atgtacaaca tcaccaaagg aacaacagcc
720acaaagggca acgagctcgg catcttcgag gacctaggag acatctacag ccaagatgac
780ctcaaccttt tcttcgccaa ctttgccagc gacatcccac agggaaccca tccaaccctc
840gactccatcg acggcgccac cgccccaaca gacgtcacca acgccggccc cgaatccgac
900ctggacttcc aaatcgccta cccaatcatc tggccccaga acaccatcct ctaccaaacc
960gacgacccca actacgaaga caactacaac ttcaaaggac tcctcaacaa cttcctctac
1020gccatcgacg gctcctattg caacgaaacc tcctctctag accctcaata cccagatccc
1080tccccaggcg gctactcctc ccccaagcaa tgcggcgtct acacccccac aaacgtaatc
1140tccatctcct acggcagccc cgaagccgac ctccccatcg cctaccaacg ccgccaatgc
1200cacgagttca tgaaactcgg ccttcagggc atcagcgtgg tcgtcgcatc gggcgactcc
1260ggcgtcgcct ccagcacggg cacctgcttt ggcgatgcag acaacgtctt cgtcccagat
1320ttcccagcca catgtcccta tctcaccgca gtaggaggca catacctccc cctaggcgca
1380gacgcagcca aggaccagga aatagcagtc acccgcttcc cctccggcgg cggcttcagc
1440aatatctacg cccgaccatc ctaccagaac cactccgtgg agacctattt ctccactacc
1500agcgacgacc tcacctaccc ttactactcc ggagtaaact acacagactt ctccaacaca
1560gatggggtat acaaccgcat cggacgagga taccccgatg tttcagctat cgcagacaat
1620atcatcatct acaaccaggg cgaagcgaca ctggtgggtg gtacgtctgc cgcggcgccg
1680gcgttcgcgg ccatgttgac gcgcattaac gaggagaggc tggcgaaggg gaagtccacg
1740gtggggtttg tgaacccggt gctgtatgaa catcctgagg cgtttaggga tgtgactgtt
1800gggtcgaatc ccgggtgtgg gactgatggg ttcccggttg ctggggggtg ggatccggtg
1860acggggttgg ggacgccgcg gtttgaggat ttgatggata tatttgtggg tgatgattga
1920931116DNAAspergillus niger 93atggcttcca agaccctcct actcattccg
gcactggcca cagccgctct gggaagtgta 60ttggacctag atatcaaggt agaccttgga
accccaggtg gcccgtttga tttgatgtac 120gacaccggat catcaacact ctgggtgctt
gatagcaatt gtacagatga ttgtccaaat 180gttagcgggt actcccgaca cggctacaac
ctcacctcta ctggtgtcaa cttaggtgtc 240aacgacagca ttgcttacag cggaggcact
gtcagcggct tcactgccac ggatattctc 300acggttcccg acaccaacgt ctcatatcgc
cagagctttg ccgtcattac cgacagtacc 360tgggcggcct tagcagccga tgggttcatc
ggcctggcat cgtctaccat cgcattcaag 420aatactacga cagccgtcga acagatgatg
caggatggac ttttggatga acctcgattc 480gccatatatg caggttcagg ggaatcgacc
gtgaccaacc ctaatccgga gaataatggc 540gtgttcacct ttggtggcag ccatgaggaa
acctatgcgg acggggaact gcaatggatg 600aagatgctct ccccctttga aatatacaaa
acaaatctcc ttggaattca gggacacaac 660aactccgatg gccaggccct gtcaagcgac
gtcctgaact ggtacggcca gactaatcta 720ttcaacgtcg caggtgcttc atcgataagc
attcccaacg accagattga ggcgatgtat 780gccctaacgc ctttctcata cgctgacatc
tcatctggat accgacctct gtgctccgat 840ttcaatgata catggtcgat ctcttttaca
atgggcttct atggcgaggg tgtcaccttc 900aatttgaccg gtgatcagct ggccgtgcct
ggctatcagg acgacgacca ctgcttccct 960cccttcaatc catgggacag ctacaacacg
attattggtc agcattggtt gagcaatttc 1020tatgctgtat tcgacttcgg atcattcgac
ccggagacat acgatatacg tgttgggctg 1080gctcctttga agaaggaata cctgccgagc
gcttga 1116941245DNAAspergillus niger
94atgtttccct gctctcgtat ttggtctctg ctcgttgcag ccgccaccgc tagtgctgta
60cccaccagtc tggccaccac gcacctgcaa tcggttgact tgcttctgac tcgcagttct
120tacgggtttc ttactgacat agcccttgga actccgggtc agagcctgcc gtatctggtt
180gactggacct ggaccggcca ctatgtggtg accaccttgt gctacaacga tcccaccgcc
240acctacgatt gtctcaacgt cgatcagaaa attttcaacc agactttgtc atccactttt
300atcaaccaaa ctgaccagta tggctatctt tactgggatc ccaaccactt ctactttacg
360gagcccgcag cagccgatgt ggcgacggac atgctgcgca tcggtcccac cgcggtgaac
420accaccatcc aagcagccaa tttcgtcttc aacgagacta ttagcgcatt ccctttctcg
480ggagtatatg gactctcacc tgtttttcag ggtgacaatc gatccgtgca agcgtccttc
540taccaaggat ggaggagcgg cgcctggcac tctccaattg tctcttttat ctactgccac
600gacaatgcca ccaaagcggt atgcagtggt tacgacggcc ttcagacact aggcggatac
660aacacctctc acgtccaggg agatatcacc tggtacgaca tcattgtcac ggaggcgatc
720aacacgctgg actttgtcta tgcgccagcc gtgattaatt attgggcgtt gaacctcacg
780cgcttctcta tcggagacga agagcaagag ctcaacaaga ccactactct ggatggaaag
840caagccgccg ttgccgcgtt cgaccacgct tcgtatggtc gcggtgcccc agtgtctgtg
900tacggttacc agcgtctagt cgagctggtc ggggcaaaag ccgtcacgct ttccgatcct
960ccaaataacg gtgagcaggg attctatcag ttcgattgcc ggaactcgag tttactgcca
1020ccgctgcggt atgagtttgc cgggtcagag cgggcgtggg agattgtgcc cgagaactat
1080gtggaggtgc tggcgaacgg aaccaataag tgcaccttta atgtacgcac cctgggagat
1140ggagcgatgg taatgggaaa ttttggcgag acatttgcca ttgataagta tgtcatgttt
1200gactttgaga agttgcaggt ggggattgca gacttcgcgt ggtaa
1245951443DNAAspergillus niger 95atgcatctcc cacagcgtct cgttacagca
gcgtgtcttt gcgccagtgc cacggctttc 60atcccataca ccatcaaact cgatacgtcg
gacgacatct cagcccgtga ttcattagct 120cgtcgtttcc tgccagtacc aaaaccaagc
gatgctctag cagacgattc cacctcatct 180gccagcgatg agtccctgtc actgaacatc
aaaaggattc ccgttcgtcg tgacaatgat 240ttcaagattg tggtagcgga aactccctct
tggtctaaca ccgccgctct cgatcaagat 300ggtagcgaca tttcatacat ctctgtcgtc
aacattgggt ctgatgagaa atctatgtac 360atgttgctcg acacaggcgg ctctgatacc
tgggttttcg gttccaactg cacgtccaca 420ccctgcacga tgcacaatac cttcggttcg
gacgattctt cgacccttga aatgacatcg 480gaagagtgga gtgtgggcta tggaactggg
tctgtcagcg gcttgctagg aaaagacaag 540ctcacgattg caaatgtcac tgtacgcatg
actttcggac ttgcttccaa cgcatcggat 600aacttcgagt cgtacccaat ggacggcatt
ctcggtctcg gtcgaaccaa cgatagttcc 660tacgacaacc caacattcat ggatgccgtt
gcagaaagta acgttttcaa gtcgaatatc 720gttggcttcg ccctttcacg tagccccgcc
aaggatggca cggtcagctt tggcactact 780gacaaggaca agtacaccgg cgatatcacc
tacaccgata ccgtcggatc ggacagctat 840tggcgcattc ccgtggacga tgtctatgtt
ggcggcactt catgcgattt ctccaacaaa 900tcagccatca tcgataccgg aacttcttat
gctatgctgc cttcaagcga ctcgaagacg 960ctgcacagtc tcattcccgg cgccaaatct
tcggggagct accacattat tccgtgcaac 1020acaactacta agctacaagt ggcattctct
ggtgtgaatt acaccatctc gccgaaggac 1080tacgtgggag caacttcagg ttctggatgc
gtttcgaaca ttatcagcta cgacttattt 1140ggtgatgaca tctggctcct gggtgacacg
tttctcaaaa atgtgtatgc tgtgtttgac 1200tacgatgagt tacgggtcgg atttgcagag
cgttcctcga acaccacctc tgcgtcgaac 1260tctacgagct ctggaacaag cagcacctcg
ggatccacta caacgggcag ctcaacgact 1320acgacgagct ctgctagctc tagtagttca
tctgatgctg aatcaggaag tagcatgacc 1380attcccgctc ctcagtattt cttctctgct
ctggcgattg cttccttcat gctttggctc 1440tag
1443961401DNAAspergillus niger
96atgacgtctt ctaccttgcg ccttgccgtc gcgttggctt tgtcaacttg cagcagtgcc
60ctatcgagcc agcgagatga ttcacttgtg gttccatttc cttttggcaa tcttgaggat
120gtccatattg ccaagcggga tagctccaag acagtagaag ctcctctagt gatatatggc
180gacagctact ggatgaacgc ctcaattgga acccctgcgc agtcactaag tttcctacta
240gatcttacgc gctcaagggt cgagcccgca tacaccctcg atgagaatta cgaatgttct
300gacgatgaac tctgctccga attcggcttc tacaaaccca ccgattcatc cacttatcag
360catctcacct acacacagag acacgatgca ggtgtcgact actcctacct tgataccata
420actcttggag atcacgcaac cgacaatgtc ccactggaca tgtatctttt gtcctacatt
480tcctacagct ccctcggtct ctcctccgtc aacaccagct tcccctacat cctggtcgat
540cgcggcctca ccacctcccc atccttcagc ctaatcggcg acaacggaaa caccaccacc
600cccagcatca tctttggagg catcaacacc tccaaattca acgggcccct gcaagccttc
660tccttcgcag accacagcat caccaacaat ccattcgtca ccgtcgaagc tgactccctc
720caactaacca ccaacaccaa cgataattcc acctacccta ttccctcctc cacccccatg
780atgctcagaa ccgaagaact aatcacctac ctccccaact cgaccgtcca atccctctac
840accgacctta acataaccat ggacggcgtg atctccactt caagattcta cggggtcctt
900ccctgcgccc gccaggaaac cgaatctcac acaatctctc tagccatcgg caacatgacc
960ttctctgtgt cctgggatga gctcttcgtc ccgtggacgc gtgacggact atgcaagttc
1020ggcattcagg cccaggattc agattacaaa actcgtgcgg agctgggtgt tccctttctg
1080agacggatgt atgtcgctgt ggattataat aatcagtttg tgggcgttgc gacgctgaag
1140gatgatgatg atcagaatgg aggtgaagat gagattgtgg agattggcac tgggacggcg
1200ttgcctagtg ctgtcgggga ttggccggct agtgttacgg cgtatacgcc tgctgcttct
1260acagggacgg cggctgcgac gttgacattc acgacggcga cgtctagcgg gggaggtgtg
1320gtgccgacgg gtctatcaga gttgggtagg gcgtttttgg tgccgggggt gctggggatg
1380gctgttttgc aggctgttta g
1401971632DNAAspergillus niger 97atgatgcgac cgatacttct ccccctactg
ggggtatttc tgcagacctc ctcggcatcc 60aatccctatg taatgagctg gtcttcccaa
gcctacggtc cagatggccc gtggcaggcc 120gtatccatcg acgtgggcag caaccagcag
acggtcgatc tttaccccgg agccaactat 180gctagcacga tcctgatgag cactctctgc
acgaacaaaa ccctgtcatc cacctgctac 240gctgccgaag caggcacgtt caaccaaaac
acctccacca ctgcctacac caccgccagc 300tcgtgggaaa caacttactg ggccgtcgag
ggtggaagcc aagaggctgt gctcggcgat 360gaggtcacct tagggtcgtt tgtcgtcccc
aatgtgagct tcgaagccat ctaccagacc 420taccagacct atcccaatgg catcgcctat
cctgtctcgg tcggcagtct ggccctgggg 480ggtccgtact tgtcggatac cgtctccaat
tcgacggtcc tgaacatgat cgcaggatgg 540ctttactcgt ccaacgacat tccgtcctac
tcgtacggca tgcatatcgg gtcggtagac 600cccaaaatcc caggctccct gatcttgggg
ggctacgata agagccgagt gatcggagac 660gtgagtgcgc agggagtagt gtcttcgagt
ggtcttttgg aacttgaatt aaaggatatt 720gggctgggtg ttgcggcggg ttcctctccc
ttcagcttca acaacgaaag tggcttgttt 780ctccaaagca gtggttcggt tcaggccaag
accgtccaga ttgatccaac caagccctac 840atgtaccttc cccaggcgac atgcgatgcc
atcacctcca ccatgccgat ctccttcaat 900tccagcttgg ggctatactt ctgggacacc
acgagcgatg attatctgaa tatcacgtct 960tccgccgcat acctctcctt tgtgttcaac
atgaatgggg tcaacaacaa gaacattacc 1020atcaagattc ccttttccca gctcaatctt
acgctgcaag aaccgctggt cgatcaaaac 1080gtcacctact tcccgtgctt cctcactacc
tccaccccgg tgctcggtcg agcctttctc 1140cagtccgcat tcgttggggt gaactggttc
aacgggaaca actcgggcac atggtttctg 1200gcacaggccc ccggcccggg ttacgccagt
gaagacatca cccggatcgc agtgagtgac 1260acgtcgcttt ctgcctctaa cggtacctgg
gaagagacct gggctacgta ctggggcatc 1320aaaacatccg acaactcgag cagctccaag
agtggcctgt cttccggtgc caaaattgga 1380attggcgtcg gggtgggtgt cggtggagca
gtgttgatcg cagcaggtat agccattgca 1440ttctgtcttc gccgtcgccg cggggcgagt
caagaggcgg ctggagagca acggaggtcg 1500atgtttaggg gctttgcgga gctaccggga
ggtgctcaca gtgaaccggc gaaggagttg 1560gatacgaaga tgcataagcc gccgcaggaa
atgatggctt cgcaggaggt agagcgatac 1620gagctggggt ga
1632982535DNAAspergillus niger
98atgcgtctta caggtggtgt cgctgcggct ctgggcctct gcgctgctgc ctccgcttct
60ctccatcccc atcgttccta cgagacccat gattacttcg ctctacacct tgatgaatcc
120acctcgccgg ccgacgtcgc ccaacgacta ggtgctcgcc acgaaggccc cgtcggagaa
180ttaccctcac atcatacctt ctcgataccc cgtgaaaaca gtgacgatgt ccatgcgctg
240ctggatcaat tgcgcgatcg tcggaggtta cgccgccgct ccggagatga cgccgctgtc
300cttccctcct tggtcgggcg agacgaaggt ctaggtggca ttctttggtc cgagaagctg
360gctccccaga gaaagctcca taaaagagtg ccgccgacag gatatgctgc cagatcgccc
420gtcaacactc agaatgaccc ccaagcgctt gcggcgcaga aacgcattgc ctcggaattg
480ggcatcgcgg accccatctt cggcgaacaa tggcatttgt ataatactgt tcagttgggc
540catgatctta acgtgacggg tatctggctg gagggcgtta cagggcaggg tgtcacgacg
600gctattgtcg atgacggttt ggacatgtac agcaacgatc ttaggccgaa ctattttgcg
660gcgggttctt atgactataa cgacaaagta ccagagccga ggccgcgctt gagcgatgac
720cgccacggta ctagatgcgc gggtgaaatc ggtgcggcga agaacgacgt gtgcggggtt
780ggtgttgcgt atgatagtcg catcgctggt attcggattc tctccgcacc cattgatgac
840actgatgagg ctgcggctat taactacgcc tatcaggaga acgatatcta ctcgtgttcc
900tggggtccct atgacgatgg cgccacaatg gaagccccgg gcaccctgat caagcgggcc
960atggtcaatg gtatccaaaa tggtcgtggt ggaaaaggct cggtttttgt gtttgcggct
1020ggtaacggtg ccattcatga cgataactgt aactttgacg gttacaccaa cagtatctac
1080agcatcacgg tgggtgccat tgatcgggag ggtaaccatc ctccgtattc ggaatcctgc
1140tcggcgcaac tggtggttgc ctacagcagc ggcgccagtg atgcaattca taccacggac
1200gtcggcacag acaagtgctc gactacccat ggtggaactt cggcggccgg cccgctcgct
1260gcgggaaccg tggcgctggc cctcagtgtg cggccggaac tcacctggcg tgacgttcag
1320tatttgatga ttgaggcggc agtgcctgtt catgaagatg atggaagctg gcaggacact
1380aagaacggga agaagttcag ccatgactgg ggatatggta aggtcgacac atatacgctg
1440gtgaaacggg cagagacctg ggatctggtg aagcctcaag cctggctcca ttccccctgg
1500cagcgggttg agcatgagat cccacagggc gagcagggct tggctagttc gtacgaggtg
1560acggaggata tgttgaaggg agccaacctg gaacggctgg agcatgtcac ggtcaccatg
1620aatgttaacc acacccgccg aggcgatctc agcgtggagt tacggagccc tgacggtcgg
1680gtcagtcacc tcagtacgcc ccggcggcca gataatcaag aggtgggcta tgttgactgg
1740accttcatga gcgttgctca ctggggcgag tccgggattg gcaaatggac tgtgattgtc
1800aaggacacca atgtcaacga gcatactggg caattcatcg attggcgact caacttgtgg
1860ggcgaggcga ttgacggagc cgagcagcct ctccacccca tgcctactga acacgatgac
1920gaccacagct atgaggaagg aaacgtggct accacgagca tcagcgccgt tcccacgaaa
1980accgagctgc ctgacaagcc cactggtggc gttgatcgcc cggtgaacgt taagcctaca
2040acatccgcga tgccgaccgg tagtcttaca gagcccatcg atgatgaaga actccagaag
2100acccctagta cagaggcaag ctcaacacca agtccttctc cgaccaccgc gtcagatagt
2160atcctgcctt ccttcttccc cacgttcggt gcgtcgaagc ggacgcaagt ttggatctac
2220gctgcgatcg gctccatcat tgtgttctgc attggcctgg gcgtctactt ccatgtgcag
2280cgccgcaaac gtattcgcga cgacagccgg gatgactacg atttcgagat gatcgaggac
2340gaggatgagc tacaggcaat gaacggacgg tcgaaccgtt cacgtcgccg gggtggcgag
2400ctgtacaatg cttttgcggg cgagagcgat gaggaaccgt tattcagtga tgaggatgat
2460gaaccgtatc gggatcgggg gatcagcggc gaacaagaac gggagggcgc agatggagag
2520cattctcgga gatga
253599450DNAAspergillus niger 99atgaagacct tctctaccgt cacctctctc
ctcgctctct tctcctcggc tctggccgca 60cccgttgaca gcgctgaagc cgccggcacc
accgtctctg tctcatacga cactgcctac 120gatgtctctg gagcttcctt gaccaccgtc
tcctgctcgg acggtgccaa cggcctgatc 180aataagggct actccaactt cggctccctt
ccgggcttcc ccaagattgg aggcgcccct 240accattgcag gctggaactc tcccaactgc
ggcaagtgct acgccctgac gtacaacggc 300cagacagtca acattctggc cattgattcc
gcacctggtg gcttcaacat cgctctggag 360gccatgaaca ccctcaccaa caaccaggcc
cagcagctgg gtcgtatcga agctacctat 420actgaggtgg atgtcagtct ttgcgcataa
450100891DNAAspergillus niger
100atggctcaaa tattctggct ttcactcttc ctgcttgtct cttgggtcag agccgagtcc
60aaccgcaccg aggtggacct gattttccca agaaatgata cctttgcgcc aatgcctttg
120atgccggttg tattcgccgt tcaagcccct tccgtcgccc ataaagttaa tacatacatc
180gagtacggct attacccagt aggccgtcca aatgaaacag ttattggcca gaccgaccat
240gtgtccgact caacaaacga aaccacttat ttcagtgtct ctggtatcgg cagaacgttc
300aataccactg gcagctggga gctgttttgg aggctgagat ggaccaattg ttcaatctca
360gaagactcga gatactacaa ccaatcctac ccctggatat cctccccata catcgacggt
420agcctcaaca tcgacaaggt ctatgagggc tttcactaca cagcatacaa tgtcattgtc
480gacagggtta ccttcagcac tcgcgaagat gctagccaac ccaacctcac gaccctcacc
540aatagcgaga actgcgataa agtctcgtct cttgctctat tgtcgattgt ggactcccta
600aggattccac cccagttacc ccaagaagat attgataccg tgtcaatgtg cccacaactc
660gccgatgcca ggctaaattc aacttcaact tcaagcccct gcagcgttag cattagtccc
720gaggttgagt ctaatatcct ggccaagatc gcagacaatg aatgcaataa cgcacttcac
780cccgctgtga gttgcaccac tgaagaaacc aaggaaggca gcgcgagcag ccatgaccac
840ggccatgctg tatggcttgt cattacgcta gcttttgcct tccttttcta a
891101933DNAAspergillus niger 101atgggtggtc gagatgtcgc cattctcagc
aggcactttg ctgtgacatc ctcacaaagt 60gttaatggcg ttgtctctgg gatgttccaa
cacacagtca cctcttcacc cagcttcact 120accaaccaat tcttcaagaa gaagttcact
gctgcaattg ctactgccat tttcgcaagc 180gttgccgtcg cagctcccca gcgtggcctc
gaggcccgcc tcaaggcccg cggcagcagc 240aagggatccc gacccctcca ggcagttgct
agacctgcat caaccaagaa ccagaccaac 300gttgagtaca gctccaactg gtccggtgcc
gtgctggtgg agcctccctc tgctgcagcg 360acctacactg cggtgaccgg caccttcact
gtccctgagc ccaccggcaa ctctggaggc 420agtcaggctg catctgcctg ggttggtatc
gacggtgata cctatggaaa cgccattctt 480cagaccggtg ttgacttcac cgtgaccgac
ggagaggcct cgttcgatgc ctggtatgag 540tggtacccgg attacgccta cgacttcagc
ggcatcgaca tctcggcagg cgatgagatt 600gttgccattg tggagtccta cacctcgact
accggtattg ccattattga gaacaagagc 660accggccaga aggtgtccaa ggagctgtcg
tccagctcca gcctcggtgg acagaacgct 720gagtggattg tggaagactt cgaggaaaat
ggttcgctcg tcaacctggt ggactttggc 780accgtcacct tcactggtgc tgttgccaag
gcggcgggtg gtgagagtgt tggacttacc 840gatgcgacca tcatcgagat tgaggagaat
ggccaggttg tcactgacgt taccatcgac 900agcgactctg aggtgaccat cacctacgag
taa 9331022046DNAAspergillus niger
102atgcgctgct ccctcatctc ccttctaggc ctggcggcca tcccggccct tggaggctgt
60cccttcgcac acactgcgaa catgggcatt gataacatgg tgaaagcaca cgctcacatg
120tcccgaccgt tgattgcctc caagagcagc ccctcaactg ttcctacctc ctctagcacc
180ccttctgtcg ggcagaaagg cgtgttcatg atgaaccgca ttgctcctgg cacatccgag
240ctctacattg ccaacacaga tggcagtaat gaacgcccac tcctctccaa ccccgtctac
300gagtaccatg cctccttctc cccggatgta gaatggatca ccttcaccag cgagcgcaat
360ggtgacggta actctgacat ctaccgcgta cggaccaacg gctccgatct ccaggaattg
420gttgccacgc ctgcagtgga agactccgtt gttatctctc ccaacggccg cctggcagcc
480tacgtctcca ccgccaacaa catgaaggca aacatctgga tccttgatct tcagaccggc
540gcgcagtgga acctcacaaa tacacccacc actgccgcca actcctccct catggagagc
600tatctccgtc ctgcctggtc tcctgatggc gaatggatcg ccttctcttc ggaccgcaac
660acccaatggg acggacacgg cgtaccgacc ttcctcggcc gcacgggctg ggagacgacg
720caagaactct ctctctacgc catccgtccc aatggctctg acttccgtca gatcatctcc
780aagccatact actctcttgg atctccgaaa tggtcagcag acggtaaacg catcgtctac
840tacgaaatga cccgggaaga cacctacaac gcccatcgtc cagaaaccat taccacagcc
900aactcgacga tcatgtccgt agacttcgag acaggcaccg atgtgcgcgt ggaagtcgcc
960ggctccggtg tcaagcaatt ccctcagtac ctggacaaga acggcaccat cgcctacacc
1020ctcaaaggcg gcaccagcga gggcttctac acgaccgcgg gactctacgt caacacgacc
1080tcggcgaccc tcaggtcccc ggcgtggtct cccgacggca agcaagtagt ctacgaaaag
1140agcacctgga gcatccgctc ggggtacaag cagctctaca gctgggacag tgactgggac
1200taccgcttca cggacgtctt ccctcaggtc tcgcaccagg agcgcgtcgc catcacacag
1260aagcagctgg gcaattcgtc catcgtgacg ttgaacacaa ccggaggcga cttgcaactc
1320gtctacgacc ccagcacggc ggactttgtc agcgatgacg aaaccacagg actgagcgct
1380taccagccca gctggtcacc ctgcggcgag tggctcgtct tcggcgtcgg attctggttc
1440gagacgagag aagcctcagg cggatggatc gtgcgggcca ccgccaacgg gagctactcg
1500gaggttctcg tgaacagcag ctactccatc accgaggatg gagccctgaa cagcgggttc
1560ccgagtttct cgccggatgg caagaaagtg gtgtatcggg tttggggagc cgacactgca
1620acctacggca acgccagcga gatcgggctg cgggtgctgg acctcgagac gcgaaagaca
1680accgtcctaa ccacagaatg ggacaatctg ccccagttct ctcccgatgg agagctcatc
1740ctattcacac gcaaaaccag cacgtacaat tacgatgtgt gcacgatccg gccggatggg
1800acagatctcc gcgtgttgac gagcagcggt gctaatgatg cgcatgcggt ctggtcgcag
1860gatggacgga ttatgtggtc taccggcatg tatgggttcc ggtttgagtg tgcgctgtat
1920ggtgatacgt tccagccgta tgggcaggtt atgattatgg atgcggatgg gggaaataag
1980aagttgatga ccaactcgat gtgggaagat tcgatgccgt tgttcttgcc gagggaggta
2040ctttag
20461031875DNAAspergillus niger 103atgcctccgg atgcaaaatc gcctggctac
cagcctggta tggcagtatt accatctagg 60ccacatcctg ccaagggaaa agccattcga
ttcctccttt cccttgcatt ggtcgcgttt 120gctattgttc aattatgtgg taatttccac
aaaaatagga gcgttgaaca acagcttcag 180agtcaaacac ttgatgatga gtcctttaaa
tgggaagatg ttactcctac caagcaactc 240gtataccatc catgctttgg tgatcacgaa
tgcgctcgct tgtcgcttcc aatgaattgg 300aaccgaactg atggtgaagg gtcaaaaatt
gccttggcgg ttatcaaact tcctgccaag 360gtacctgtca cagatgcgcg atatggtggt
gccattcttc tgaatccagg tggtcctggt 420ggatccggag tgagcatggt ctttagatac
gggaaagcta tccagaccat cgtcgactcc 480ccagaatcac caagtgcaga ttcagcgagc
ggaaagtatt tcgatgttgt tagctttgat 540ccaagagggg tcaacaacac aacacctaat
ttttcctgct tccctgaccc cgcgacgagg 600aaagcgtggt tactgcagtc agaggcagag
ggtctacttg ggagttctga aggagtcttc 660gatactcgat gggcaaggta cgaagctttt
gagcggctac tttcgacagc tccgaacact 720ttcccagttg gaacaaacgt tgacgccgag
aggataaggc tgcacaaccg ttggaaaaaa 780ggggaggaga agctgctata ctggggcttt
tcctatggga caatcctggg ttccacgttt 840gcggctatgc agcctcatcg cataaaccgt
gctgtcatag acggagtctg caacgctgat 900gattattacg ccggcaactg gcttaccaat
ttacaagatt cggatgcagc attcaataaa 960tttttcgagt actgctacac agctggccca
tcagcgtgtc cgtttgcgct cggcggagat 1020cccgaagatc tcaagtctcg ttatgagcag
attttgacca atcttacatc gagccctatt 1080gctgtgtctc cttctggaaa taggggccca
gagataataa cctatagtga tgtgaagtca 1140ttggtcgtgc aagctctcta tgtgcctttg
aaattattcg atttggtggc taggctatta 1200gctgagctcg agcaaggtaa cggctcttca
ttcgctgact tgaagtatga agccaaacaa 1260tggccagtac cgcctccatg cgattcctcg
tccacacaat acaaagtacc tggcgagagt 1320gatcaggagg ccgggaggaa tatcctatgt
acagatggtc caggcctcga cggaactgcc 1380aaggaggatt tccggagcta ctggaatatg
ctccggggac aaagtaaggc ggttggagat 1440ttctgggccg aggttcgcat gtcgtgtgtc
aaactggaga cgcgacctga gtggcgctat 1500gatggtatgc gtatccaagg gcccttcgca
ggcaatacat cgcacccatt gctgtttatc 1560gggaatactt atgatccagt aacgccgcta
cggaatgctc atacgatggc gcgtggattt 1620cctgagtcaa tcgttctaga gcagaactct
gtcggacatt gcacactgag tggcccatcc 1680ttgtgtacag cgaaagcgat acgccagtat
ttccagaccg gagagttacc tgaccccgga 1740actgtttgcc aggtagagga gcttcccttt
cgtcttgccg gatatgagag aagtcaggtc 1800atgtcgccag gtgacacaga attgatgtcc
gccttgcatt cgctgagcga gttccgccat 1860ctgctaggcg cgtga
18751041665DNAAspergillus niger
104atgttgagta gtctgctgct tgggggtctt ctgggtctag cgaccgctca atttcctccc
60gagccggaag gcatcactgt gctcaagtcc aagttgcatg agaatgtgac tatttctttc
120aaagagcctg gaatttgcga aactacgccg ggtgtccgat cttattcggg ctatgtacac
180cttccccccg cctcaaccag cttcttttgg tttttcgaag cccgcaaaga tcccagcaat
240gcgcctctgg ccatctggct caatggcggt ccgggtggct cgtcgctcat ggggctcctt
300gaagaattag gtccttgttc cattgcatca gactccaaga ccacagtcct caatccttgg
360agttggaaca atgaagtcaa tcttctattc cttgaccagc caactcaagt cggcttctca
420tacgatgtcc caacaaatgg cactttgaca gctaatggga ctgcattcgc ggctcacgct
480ctatggcatt tcgcgcaaac ctggtttttc gagttcccac actacaagcc aaacgatgat
540cgtgtcagtc tctgggctga aagttacgga ggccattatg gtccaggcat ctttcggttc
600ttccaacagc agaatgacaa aatcgcagag gggactgcag aagacggtgc acagtatttg
660catctcgaca cgcttggcat tgtgaacggc ttgatggata tggtgatcca agaagaggct
720tacattactt ggccatacaa taacgtaagg ctcgcccctt cttcattcaa ctcgcgaggc
780tttcgcgatc aggccctcgc ctgcgaagcg gctttgaaag aacgcgattc cggcttgcct
840cactcaggga agaatatctc tgaaatttgc ggaggccttg cactagaatg gggagatggc
900cccatcacct actaccacac cttcaatcgc gggtggtacg acatcgccca tcctaagaac
960gacccattcc ctgccaagca catgctcgga tatttgacgc aggagtccgt ccttgccgct
1020cttggggtac cagtcaattt cacatcgtct tcgagtgccg tggctacaca gttcataaaa
1080acctttgata tcgtccacgg cggcttcctg gatgcaattg gctacctcct cgacagtggt
1140gtaaaagtac acatgatgta cggagatcgt gattacgcct gcaattgggt cgggggcgaa
1200aaagccagcc ttgcagttcc gtattcccgt atcaccgaat ttgccgacac gggatactcc
1260ccactcctta cgcccgacgg gatcagcggc atgacccgcc agctgggcaa ctacagcttc
1320actcgcgtct tccaagccgg gcatgaggtc ccctcctacc agcctgtcgc ggcgtatgag
1380atcttcatgc gggcgacatt caacaaagat atccctactg gcctcttggc tgttgatgac
1440gaattccagt cggttggacc taaggatacg tggcatatca agaatatccc tcctattatg
1500ccaaagccgc agtgctatgt tctaagtccc ggcacgtgta ccccggaggt ttgggagacc
1560gttttgaacg gatccgcgac ggtaaaggat tggtatgtcg tggatgatag cgcgggtgtt
1620gaggaccacg aggggttcag cattcttgga ggggatgagt tgtag
16651051737DNAAspergillus niger 105atgaccaggt ttcaattgct tccccttgtc
gcagggctgc ttgccccttc aattgcagcc 60cttagcatcc cttccccgca gcagatcctc
gattctctca ctttcggaga gcacaccgac 120ggcttttgtc cgctggcacc caaggttgag
gttcctgacg atggtttctt tccagctctc 180aagttcgtag aagatgcctc gttcaagtcg
cgccaagtca atcgtctctc cagggcggtt 240caagttccga ccgcaatcga cgactacatg
aaggatccct acgacgaaaa gttcgcccca 300ttcctcgact tccagaagct cctgcagacc
ctctttcccc tcacccactc ctacgcccgc 360gtagatcaca tcaaccgatt tggtctcgtc
ttcaccctca atggcacaga tgactcgctc 420aagcccctgc tattcaccgc gcaccaggac
gtcgtgccca tcaacgaccc tgccgactgg 480acctatcccc ccttcgatgg ccactacgac
ggcgaatggc tctggggccg cggtgccagc 540gactgcaaga acgtcctgat cggtctcatg
tccgttgttg aagacctact ctcccaaaag 600tgggagccaa cccgcacagt cgtcctggcc
ttcggattcg acgaagaatc ccacggcttc 660ctcggcgccg gatccatcgc caaattcctt
gagaagaaat acggaccgga cagcttcgaa 720tttatcctcg acgaaggcgg catgggcctc
gaagttctag acgacaacaa caacggcgtc 780gtctacgctc tccccggcgt tggcgaaaag
ggcagcatcg acgttgtgct cactctggcc 840gtaccaggcg gccacagctc cgtgccccct
ccacacacgg gaatcggcat catcgccgag 900atcatctatg agctagaacg ccaggacctc
ttcgtccccg tcctagacac tcaccacccg 960acccgcaaga tgctcgaatg ccaagtccgc
cactccccct cgcaagtcga accgtggctc 1020gcctccgccc tccaatcaag cgactacatc
tccctagcag agaaactggc ctcctcgcgc 1080ggcgacaagt tccgcttcat cctccaaacc
tcccaagcag cggacatcat caacggcggc 1140gtcaaatcca acgctctccc cgagaaaatc
aacgccctcg tcaactaccg catcgctctg 1200caccaaaccc cagacgatat caagaaccgc
gctgtggaga tcatctctcc catcgtcaag 1260aaatataacc tctccctcac ggccttcccg
gaaagcgaca ccgttgaccc ctccctcaac 1320aaccacctca cccttactac cctcagcggc
gccctcagtc ccgccccggt cagcccaacg 1380gacatcgaca ccgacgccgt ctgggcccgt
ttctcgggcg tcactcgctc ggtcttcgaa 1440tctgtcccta gtctcgaggg cagaaaggtc
gtcgtgagcg gcgacatcat gaccgggaat 1500acggatacga gattctactg ggctttgtcg
aggaatattt acaggtggag tccgtcgagg 1560gcgggtaaag cgctgaatat tcatactgtt
gatgagagga tcgatattga tattcatctt 1620gaggcgatga tgctgtatta cgatcttatt
cgctctttcg atggacggac cgattcatct 1680gtcatttctg ctgcgtcggc agctgctgat
gatgaacttg ctcacgacgt gctgtga 17371061371DNAAspergillus niger
106atgaagagca ccactcttct ttccttggcc tgggctgccc agtccgccta ttccctctct
60atccacgagc gcgatgaacc cgctactctt cagttcaact ttgaacgtcg tcagatcgcc
120gaccggtccc gtcggaagcg atcgacggcc tcggccgacc tcgttaacct ggctacgaat
180cttggctaca cgatgaacct cacactcggc actcccggcc aggaagtcag tgtgacgttg
240gacaccggca gcagcgatct ctgggtcaat ggggccaact cgtccgtctg cccctgtacc
300gattacggct cttacaactc aagcgcttct tccacctaca ccttcgtgaa cgatgagttt
360tatatccagt atgtcgacgg cagtgaagcc acaggcgact atgtcaacga tactctaaag
420ttctccaatg tgactttgac gaactttcaa tttgccgtcg catatgacgg cgactccgag
480gagggggtcc tcggtatcgg atacgccagc aatgaagcca gccaggccac cgtcggtggt
540ggtgaataca ccaacttccc cgaagccctc gtcgatcaag gcgcgatcaa ctggccggcc
600tacagtctat ggctcgatga cctcgacgaa ggaaaaggca ccattctgtt cggcggagtc
660aacaccgcca agtactacgg cagcctgcag accctgccta tcgtctccat cgaagacatg
720tacgtcgagt tcgcggtcaa cctgacggcc gtgcaccttg agaagaacgg caactccgtc
780tcggtcaaca acagcgccac gcaattcccc atccccgccg tgctggacag cggcacggcc
840ctgacctaca tcccgacctc cgccgcagcc agcatctacg aggccgtcgg tgcccaatac
900ctgagcgagt acgggtacgg agtgatcgag tgcgacgtca aggacgaaga cttcaccttc
960ctgttcgact ttggatcctt caacatgagc gttgacatca gcgagatgat cctcgaggcc
1020agttccgaca tgaccgacat gaacgtttgt acgtttggcc tcgcagtgat cgaaaatgag
1080gccctgctgg gcgatacctt cctgcgcagc gcatacgtcg tctacgatct cggaaacaac
1140gagatctccc tggccaaggc caacttcaac cccggcgagg accacgtcct ggagatcggc
1200accggatcgg atgccgtgcc caaggcgacg ggggcgacgg cgaccggcgc ggcagccaca
1260tccacggcct cgagcgacaa gtcggacaag gagagttcgg ctacagtgcc gcgcagccag
1320attgtctcgc tggtggcggg agtcttggtc ggtgttttct tggttctgta a
13711071995DNAAspergillus niger 107atgttggtcc gtcagcttgc cctggctctg
gccattgcgg ccttgtccga tgccattccg 60acatccatca agcatgtcct gcacgagaaa
cgtcacaagc ccgcatccga ctgggtgaag 120ggtgcgcgcg ttgagagcga tgcggtcctg
cctatgcgca ttggccttgc ccagaacaac 180ttggacaagg gctatgactt cctgatggaa
gtatcggacc ccaagtcttc caaatacggc 240cagtactggt cggcagacga ggtgcacgac
atcttttcgc catccgagga ggctgttgag 300gcagtgagag aatggcttgt cgcctctggt
atccatccgt cgcgggtggt gcactccgac 360aacaagggct ggctcgcgtt cgacgcctac
gcccatgaag ccgagaggct gttcatgacg 420gaattccacg agcacgagag cgaccgaagt
gctaagatca gggttggatg cgaccaatac 480cacgtccccg aacacatcca gaagcacatc
gactacatta cccctggagt gaagctcacc 540caggtcgtga agaggaccaa caaagtcaag
cgtgcttccc aactagctca ctcttccaag 600gccaagtctg ctgcccaagg tccgcagcca
ctccccaaca aggccaagtt cctgcctgaa 660gacctccgcg gctgcggtta caacatcacc
ccctcgtgta tcaaggcctt gtatcagatc 720ccagacgcta agacggcgac cccgaacaac
agcctgggtc tgtacgagca gggtgactac 780tttgccaagt ccgacctcga cctcttctat
aaggagtatg cgccgtgggt tccccagggt 840acctatccca tcccagccct gattgatggc
gccaattact cggttccttc ctacagctcc 900ctgaacacgg gtgaatccga cattgacatt
gacatggcct actccctgct ctaccctcag 960caggtgaccc tctaccaggt tgacgaccag
ctctacgaac cagtcgaggt cgacacaacc 1020aatctgttca acaccttcct cgacgctctc
gatggctcct actgcaccta cagcgcctac 1080ggcgagaccg gcgatgaccc gtcgatcgac
cccgtatacc ccgacacccg ccccggcggc 1140tacaaaggaa agctccagtg cggcgtctat
aagcccacta acgtaatcag cgcctcctac 1200ggccaatccg aagccgacct ccccgtcagc
tacaccaagc gccaatgcaa tgagttcatg 1260aagctcggtc tacagggaca ctccatcctc
ttcgcgtctg gcgactacgg cgtcgcgtct 1320ttcgccggcg acggtgacga gaacggctgt
ctcggcccag agggcaagat cttcaacccc 1380cagtacccct ccaactgccc ctacgtcacc
tccgttggag gtaccatgct gtacggctac 1440cagaccgtca acgacagcga gagcgtcatg
cacgttaacc ttggcggaac cgcaagtaac 1500ttcagcactt ctggtggctt ctcgaattac
ttcccccaac cggcatatca gtttgctgct 1560gtggagcaat acttccagtc tgcgaacctg
tcgtatccgt attactcgga gtttgaggtc 1620gatgttaaca cgaccaaggg tctctacaat
aggcttggtc gtgcttatcc ggatgtctcg 1680gcgaatggag cgcatttccg cgcttatatg
gatggatacg attatcattg gtatggatcg 1740agtttggcgt cgcctttgtt cgcgtcggtt
cttactttgc tcaacgagga acgcttcgct 1800atcggcaagg gccccgtggg attcgtgaat
cccgtgcttt atgcttatcc gcaagtgctg 1860aacgatatca ctaatggtac taatgctggg
tgtggaactt atgggtttag tgctattgag 1920ggatgggatc ccgctagtgg tttgggtacg
cctaactacc cattgatgaa ggagctgttc 1980ctctctttgc cttag
19951081563DNAAspergillus niger
108atgcgggtta ccacggcaat tgcttcatta ctactggtcg gctcggccac cagtctccaa
60aatcctcatc gtcgggctgt tccgccccct ctctcgcatc gcagcgtagc gtctcgctcc
120gtgcccgttg agcgccgaac caccgacttt gagtatttga ctaacaagac tgcaagattc
180ctggtcaatg gcacaagcat ccccgaagtc gatttcgacg tcggcgagtc ctacgccggc
240cttctcccca atacgcccac tggcaattct agcctattct tctggttctt cccctcgcaa
300aatccagagg ccagcgatga gatcaccatc tggctcaacg gcggccccgg atgtagctcc
360ctagacggcc tgcttcaaga gaacggccca ttcctctggc agcctggcac ttacaagccc
420gttcctaatc catactcatg gaccaacctc accaatgtgg tttacatcga ccaacccgcc
480ggcacaggct tctccccggg cccctcgacc gtaaataacg aggaagacgt ggctgcccag
540ttcaacagct ggttcaagca cttcgtcgac accttcgacc tgcacggccg caaggtctac
600atcaccggtg aaagctacgc gggcatgtac gtcccctaca ttgccgatgc catgctgaac
660gaggaggata caacctactt caacttgaag ggtatccaga tcaacgaccc gtccatcaac
720agcgactcgg tcatgatgta ctcccccgcc gtccgccatc tgaaccacta caacaacatc
780ttccagctaa actccacttt cctctcctac atcaacgcca aagccgacaa gtgcggctac
840aacgccttcc tcgacaaagc catcacctac ccacccccca gtcccttccc caccgcccct
900gaaatcaccg aagactgcca agtctgggac gaagtcgtca tggccgccta cgacatcaac
960ccctgcttca attactacca cctgatcgac ttctgcccct acctctggga cgtgctcggc
1020ttcccctccc tcgcctccgg cccaaacaac tacttcaacc gctccgacgt ccagaagatc
1080ctgcacgtcc ctccaacgga ctactccgtg tgctcggaga ccgtcatctt cgcgaacggc
1140gacggcagcg accccagctc ctggggtccc ctacccagcg tcatcgaacg cactaacaac
1200actatcatcg gccacggctg gctcgattac ctcctcttct tgaacggctc gctcgccaca
1260atccagaaca tgacctggaa cggtaagcaa gggttccagc gtcctcccgt ggaaccgctc
1320ttcgtccctt accattatgg tctggctgag ctgtactggg gcgatgagcc tgacccgtat
1380aaccttgatg ctggcgctgg atacctgggt acagcgcata ccgagcgcgg gttgactttc
1440agctcggtgt atttgtctgg tcatgaaatc ccgcagtatg ttcctggtgc ggcttaccgc
1500cagttggagt tcctgctcgg taggattagt agtctttcgg cgaaggggaa ctatacctct
1560tga
15631091656DNAAspergillus niger 109atgcgtggct ctcggttggt gctcttgttg
cccctggctg cacttagttg tgctatgccc 60gagaatgaat ggtcatctac gataagaagg
cagttaccaa aagcgtccac tggcgtcaaa 120tcgataaaaa ccccaaacaa tgtcactatc
aggtataaag aaccaggaac cgaaggaatt 180tgtgagacaa cacctggggt caaatcatac
tccggatatg tcgatctttc gccagagtcg 240catactttct tttggttttt cgagtcacgc
cgtgaccccg aaaatgatcc agtgactctg 300tggctgaatg gtggccctgg aagcgattcc
ttgattgggc tttttgaaga gttgggtccg 360tgtcacatca caccagagta cgaatcaatc
atcaatcagt actcctggaa cgaggtcacc 420aatcttcttt tcttgtctca gcccctcggt
gtggggttct cttacagtga aaccgaggcc 480gggtccttga atccatttac tggagccgtc
gagaacgcct cctttgctgg agttcagggt 540cgatacccag ttattgatgc cactatcatc
gacacgaccg atatcgctgc acgcgcaacc 600tgggaggtgc ttcagggctt cctcagtggc
ctgtcgcagc tagattccga agtcaagtcc 660aaggagttca acctgtggac agagagttac
ggaggacact atggaccagc gttcttcaat 720catttctacg agcaaaattc gaagatcgct
agcggggaag tcaatggcgt ccaactgaat 780tttaactccc tcgggattat caacggcatc
attgatgccg cgattcaggc agactactac 840gcagactttg ccgttaataa tacatatgga
atcaaagctg tcaatgacac agtgtacaac 900tatatgaagt tcgccaacac gatgccaaat
ggatgccagg atcaggttgc ttcgtgtaaa 960ttgaccaata ggacctcgct ttctgattat
gctatatgta cagaagcagc caatatgtgc 1020agggacaatg tcgaagggcc ttactaccag
tttggcggcc gtggcgtgta tgatattcgg 1080cacccctaca atgacccgac cccgccgtcc
tactttgttg actacctcaa gaaagactca 1140gtcatggatg ctatcggcgt ggacattaac
tacaccgagt ccagcggcga agtatattat 1200gcattccagc agaccggcga ctttgtatgg
ccgaatttca ttgaggacct cgaagagatc 1260ctccaactcc ccgtacgcgt gtcgttgatc
tacggcgatg ccgactatat ctgtaactgg 1320ttcggcggtc aggccatctc actcgcagtt
aactaccccc atgcagctca gttccgtgca 1380gcgggataca cacccatgac agtagatggg
gtcgaatacg gtgagactcg cgagtatggc 1440aacttttcgt tcacccgcgt atatcaggct
gggcacgagg ttccatacta tcaaccgatc 1500gcagcgttgc agctgttcaa ccgtacttta
tttggatggg atattgcagc gggtacaact 1560cagatttggc ccgaatatag caccaacggg
acatcgcagg ctacacacac ggagtcgttc 1620gtgccactgt ccacggcgtc gagtaccgtc
aattag 16561101872DNAAspergillus niger
110atgccttttc ccttttcgtc cgctcttctc ggctatatct taactacgag cactactctc
60acctccctag tcgcaggaca gtattaccct ccgacgcctg aggatctcac cgttattcat
120tcggagatat tccctggtgc gaggatctcc tataagcaac ccctcggcat ctgcaccacc
180accccctcca cccccagcta ctccggctac atccacctcc ccccacacac ccttaccaat
240ctctccattc caggaatcag catctcgcaa ccatacccta tcaatacctt tttctggtac
300tttccttccc gccatcacca caacaatgat acatccccac tcaccatctg gatgaacggc
360gggcccggcg gatcctccat gattgggcta tttcaagaga acgggccatg tactgtgaat
420acggactcga attccacggc ctataatccc tggtcgtgga atgagtacgt cgatatgttg
480tatattgagc agccggtgca gacgggattt agttatgatg tgttgaggaa tgggacgtta
540gatttgaatg agacgttttt ggtggggacg ttgccgagtc aggatgtgca tgggacggtg
600aatgggacgg ttaatggggg aagggcgctt tgggttgcgt tgcaggtttg gttgggtgaa
660ttctctgaat atgtttcttc tgttgacggg aatggtggtg gtgatgacag ggtgagtata
720tggacggagt catatggggg acggtatgga ccggcataca cggcgctctt tcaggagatg
780aatgagagga ttgagagtgg ggaggtaagc accgggaaga agatccattt ggatacgctg
840ggcattatca atgggtgtgt ggatttactc gtgcaggtcc cttcgttccc tgagcaggcg
900tataacaata cgtatgggat cgagggaatc aatcgcacgc tctacgaccg ggctatggat
960agttggagca agcctggcgg gtgcagggat atgatcatcg agtgtcgcga tgctggcgag
1020ctcggagatc ccctcatcat ctgcgaggag gcgtcggact actgttcgcg ggagatcaag
1080agcctgtata cgaatacctc cgggcgagga tactacgaca tagcgcattt cacgccggat
1140gcagctctcg tgccttactt cgtcgggttc ttgaatcgcc catgggtgca aaaggcactt
1200ggggtcccgg tgaactatac catgtcgtca gaggcagtgg ggaacagttt cgcctcgacg
1260ggcgattatc cgcgaaatga tccccgcgga atgatcgggg atattggata cttgcttgac
1320tccggtgtca aggtggctat ggtatatggg gaccgggact atgcttgtcc gtggcgcggc
1380ggggaagatg tcagcctgct ggtggagtac gaggatgcgg agaagttccg tgctgctggg
1440tatgccgaag tgcagacgaa gtcatcctac gttgggggtc tagtaaggca gtatgggaac
1500ttctcgttca cgcgtgtctt tcaggcgggc catgaggtgc cattttatca gcccgaaacg
1560gcgtatgaga tttttaatcg cgctcagttt aattgggata ttgcgacggg aggcatttct
1620ctggagcaga atcagagcta tgggacggag ggaccgtcgt caacgtggca tatcaaaaac
1680gaagtgccgg agagccctga gccgacgtgc tatttgttgg cgatggattc gacttgtacg
1740gatgagcaga gggaacgggt gctgagtggg gatgcggtgg tgagggattg ggttgttgtt
1800gatgatattg aggctgaaag ctcgttcagc ggtgttggtg atcagctggc acaggtccct
1860ttgggacatt ga
18721111320DNAAspergillus niger 111atgagaacat ctactctttt gctcctctgg
agcactgcag gagcagcttt ggcttctccg 60tacccgcttc ccgactcgca agtagtcttc
gccgcggatc acgaggtccc gaatacacag 120ggcaaacacg tcgtggacga ggccatactc
tcggcgctga acgctcattc tgacccagtc 180gctgcaatgg tgtctctacg tcccgaaact
gcagcttttc tagctgaacc tcgtctcttg 240cacattcggg gcgaagagaa ggcggaatgg
atgaccgaag gcgacaagct gcgcctccgc 300caacgcggaa agaagttcat ggacattacc
gagcatcagg acttctacgc agagcaggcg 360atggcttcgt ttgctgggga tcctaatctt
cccaagctgt cccataaagg tctcgtcaag 420ccgctgttct ctcaaatcga gacggaacga
atgcacgata tcctgcagca catgacctcc 480tactacaatc gatactacgg tgattatcac
ggcgagatga gctccgaatg gctgcacgac 540tacattgctg cgatcatctc caaatcgcct
ttccgcaccc acatctctct cgaatacttc 600acccatcctt tccgccaatc ttcaattatt
gcacgcttcg agcctaaagt tcgcagcttc 660tcccaacctt tgaccatcat tggtgcgcac
caagattcgg ccaattatct ttttcccctg 720ctgcccgccc ctggcgctga cgatgactgt
tccggcactg tcagtatcct cgaggccttc 780cgcgttctgg cggagaatgg ctacacgccc
aaggacgggc ctgttgaatt ccattggtat 840gcggctgaag aggccgggct actgggcagc
caagccatcg cgcggtacaa gaaggagcag 900ggcgctaaaa ttgatgccat gatggagttt
gatatgacgg cttttattgc ccgtaacgcc 960accgagacca tcgggtttgt tgcaacccaa
gccgatgcag cgctcacaaa ctgggccctc 1020aacctcagtc gagaatacat ctccattccg
gcggaagtct atgaacttgg ccccaacgct 1080ggatccgact acatgtcata cactaagctc
aactaccccg ctgcctttgc atccgaaggc 1140aacccgctcg ctgggggctc tttcccgggt
gaaatggacc cctacgtaca cggcatcaag 1200gataggatgg acgttgacga tgaaacgggc
gtcttctcta tcgaacacat ggctcggttc 1260tccgagttgg ctatcgcatt tgttgtcgag
caggctgggt gggataatac atggcggtag 13201121581DNAAspergillus niger
112atgcgttcct tctccgttgt cgctgccgcg tcactggcgc tctcttgggc gtctctggcc
60caggctgctc gcccccgtct tgtgcccaag cctatctctc ggccagcttc gagtaagtcg
120gctgcgacta cgggtgaggc ttattttgag cagctgctgg accatcacaa cccggagaag
180ggaacgtttt cccagcggta ctggtggagt actgaatact ggggtggacc tgggtcaccg
240gtggtcctct ttaaccctgg agaggtctct gccgatggct atgaggggta tctcaccaac
300gatactctca ctggtgtcta tgcgcaggag atccagggtg ccgtcattct cattgaacac
360cgctactggg gcgactcttc gccttatgag gtgctcaatg ccgaaacact tcagtatctc
420acactggatc agtccattct ggacatgacc tacttcgccg agacggtaaa gctgcagttc
480gataatagca gccgcagcaa tgcgcagaat gctccctggg tcatggtcgg tggctcatac
540agcggtgcct tgacggcttg gaccgagtct atcgcgcctg gaacgttctg ggcttaccat
600gccaccagtg cgcctgtgga ggctatctat gacttttggc aatacttcta ccccattcag
660caaggtatgg cacagaactg cagcaaggat gtgtctctgg tagccgagta tgtcgacaaa
720attgggaaga atggaactgc caaggaacag caggagctca aagaattgtt tggtctggga
780gctgttgagc attacgatga ctttgccgct gtcctgccca acggaccgta cctctggcaa
840gacaacgact ttgtcacagg atactcttcc ttcttccagt tctgtgatgc tgtcgagggt
900gtcgaagccg gcgcggcagt gacccccggc cccgagggcg tcggacttga aaaggccctg
960gccaactacg caaactggtt caattcaacc atactcccta actactgcgc aagctacggc
1020tactggaccg acgaatggag cgtcgcctgt ttcgacagct ataatgcctc gagccccatc
1080ttcaccgaca cctccgtggg taaccctgtc gaccgccaat gggaatggtt cctctgcaac
1140gagcctttct tctggtggca ggacggtgcc cccgagggaa cctccactat tgtgccccgg
1200ctcgtcagcg cctcctactg gcaacgccaa tgcccgctct acttccccga agttaacggc
1260tacacgtacg gcagcgcgaa gggtaaaaac tccgctacgg tgaacagctg gacgggtgga
1320tgggatatga cccgcaacac gacgcggttg atctggacga acgggcaata tgacccctgg
1380cgcgactccg gtgtgtcgag cactttccgg cccggtggtc cgctggttag cacggcgaac
1440gaacccgtgc agattattcc gggcgggttc cattgctcgg acttgtatat ggaggattac
1500tatgcgaatg agggtgtgag gaaggtggtt gataatgagg tgaagcagat taaggagtgg
1560gtggaggagt attatgcttg a
15811131275DNAAspergillus niger 113atgcagctcc tccagtccct cattgttgcc
gtttgcttca gctacggcgt cctctcctta 60ccccatggcc cgtcaaacca gcacaaagca
cgttccttca aggttgaacg ggtccgtcgt 120ggaaccggtg ctctgcatgg gcccgctgct
ctccgcaaag cataccggaa gtacggaata 180gctcccagca gtttcaacat cgatctggca
gactttaaac ccattacgac aacccatgct 240gctgctggga gcgagattgc agagcctgat
cagactggcg ctgtcagtgc tacttccgtc 300gagaacgatg ccgagttcgt ttcgcctgtt
cttattggcg gccagaagat cgtcatgaca 360tttgacactg gttcttctga cttttgggtg
ttcgatacga atctcaatga aaccttgacg 420ggacacacgg agtacaaccc ttcgaactcc
tcgaccttca agaagatgga cggatacacc 480ttcgatgtct cgtatggtga cgactcgtac
gcctctggcc ccgtcggaac ggataccgtc 540aacattggcg gcgccattgt caaggagcaa
gccttcggtg tccccgacca ggtatcccag 600tcgttcatcg aggacacgaa ctccaacggc
ctggtcgggt tgggcttttc ctccatcaac 660accatcaaac cggaggcgca agacacgttc
ttcgccaatg tcgcaccaag tctggacgag 720cccgtcatga ccgcctcgct caaggctgac
ggagtgggcg agtacgagtt cggcacgatc 780gacaaagaca agtaccaggg caacattgcc
aacatcagcg tggactcatc gaacggatac 840tggcagttct ccactcccaa gtactccgtg
gcagacggag agctgaagga cattggaagc 900ttgaacacct cgatcgcgga caccggtacc
tcccttatgc tgctggatga agacgtggtt 960actgcctact atgcgcaagt tcccaactcg
gtctacgtga gcagtgccgg tggttacatc 1020tacccctgca acaccactct tcccagcttc
tcgcttgtcc tcggcgagtc gagcctggcc 1080acgatccccg gtaacctgat caatttctcc
aaggttggca ccaacaccac caccggacag 1140gccttgtgct ttggcggcat tcaatccaac
ggaaacacct cgctgcagat tctgggcgat 1200attttcctga aggccttttt cgttgtcttc
gacatgcgcg gcccctcgct tggtgttgcc 1260tctcccaaga actag
12751141647DNAAspergillus niger
114atgcgcattg actccgcggc gctacatctg gtcccagtcc tcctgggcca ggtcggtgct
60ttacaattac ccttggtcca agactccaat tcacagtggc agaaaccaaa tgcaggtgat
120aaacccctaa ttagctctcc gttgcttcaa gagcaggtca aggcggagaa tctgttggac
180agggcccggc agctttacaa gattgcggag ctgggagaag acgagtataa ccaccccact
240cgcgtcattg gcagtaaagg tcaccttggc acgctcgact acatatactc cacccttacc
300gacctcggtg attattatac tgtcgtcaat cagtccttcc ctgccgtgag cggtaatgtc
360ttcgagtctc gccttgtcct tggtcacgat gttcccaagt cagctacacc aatgggtctc
420actcccccaa cgaggaataa ggagccggta tatggctccc tggttgctgt atccaacctc
480gggtgtgagg cctcggacta ctcgtccaac ttgaaaggcg ccgttgcatt tatcagtcgg
540ggaagctgtc cgttcgggac caagtctcaa ttagctggta aagcgggagc tgttgctgcc
600gtcatctaca acaacgagcg gggtgaccta agcggaactc taggaaaccc aacccccgat
660catgttgcta cctttggtat ctcagacgag gatgctgccc cagtcctgga gaagttgaat
720aaaggcgaga aggtggacgc tatcgcctac gttgatgcga tagtagagac catccacacc
780accaatatca tcgcgcagac cacggatggt gacccgaaca attgtgtaat gctgggtggc
840cacagtgaca gcgtggccga gggcccgggt atcaatgacg acgggtccgg tactctgacc
900cttttggagc ttgccacatt gctcacccag ttccgtgtca acaactgcgt gcgatttgct
960tggtgggccg ccgaggagga aggccttctc ggatctgact attacgtgtc cgttctcaca
1020ccggaagaga accgcaagat ccgcttgttc atggactacg acatgctcgg ctcgccgaac
1080tttgcgtacc aagtttacaa tgccactaat gctgtgaacc ccgagggatc tgaggagctt
1140cgtgatctgt acaccgactt ttacgaagat catgggttca actacacgta cattccgttt
1200gacggacgca gcgactatga tgccttcatt cggcatggta tcccgggtgg tggcattgcc
1260acgggagcag agggtatcaa gactgtcgag gaagcggaca tgtttggtgg ggttgctggc
1320caatggtatg acccgtgtta ccatcagatc tgcgatacgg tggccaatgt gaacttgact
1380gcgtgggagt ggaacaccaa gctcgttgcc cactccattg cgacttacgc caagtccttt
1440gacggattcc cggaacggtc cgatgaaccc atcagccctg ctgcttttga ggaaccgaag
1500taccatggcc acgcgttgca attacttcgc ggtaatacta cagggaccca gagcgtcctg
1560tggggagccc aaatccagaa tggaacagct gcatcggtgc ttaatctatt gtccatacga
1620cgcagaggca ctttcagtct aagctaa
1647115480PRTAspergillus niger 115Met His Leu Pro Gln Arg Leu Val Thr Ala
Ala Cys Leu Cys Ala Ser1 5 10
15Ala Thr Ala Phe Ile Pro Tyr Thr Ile Lys Leu Asp Thr Ser Asp Asp
20 25 30Ile Ser Ala Arg Asp Ser
Leu Ala Arg Arg Phe Leu Pro Val Pro Lys 35 40
45Pro Ser Asp Ala Leu Ala Asp Asp Ser Thr Ser Ser Ala Ser
Asp Glu 50 55 60Ser Leu Ser Leu Asn
Ile Lys Arg Ile Pro Val Arg Arg Asp Asn Asp65 70
75 80Phe Lys Ile Val Val Ala Glu Thr Pro Ser
Trp Ser Asn Thr Ala Ala 85 90
95Leu Asp Gln Asp Gly Ser Asp Ile Ser Tyr Ile Ser Val Val Asn Ile
100 105 110Gly Ser Asp Glu Lys
Ser Met Tyr Met Leu Leu Asp Thr Gly Gly Ser 115
120 125Asp Thr Trp Val Phe Gly Ser Asn Cys Thr Ser Thr
Pro Cys Thr Met 130 135 140His Asn Thr
Phe Gly Ser Asp Asp Ser Ser Thr Leu Glu Met Thr Ser145
150 155 160Glu Glu Trp Ser Val Gly Tyr
Gly Thr Gly Ser Val Ser Gly Leu Leu 165
170 175Gly Lys Asp Lys Leu Thr Ile Ala Asn Val Thr Val
Arg Met Thr Phe 180 185 190Gly
Leu Ala Ser Asn Ala Ser Asp Asn Phe Glu Ser Tyr Pro Met Asp 195
200 205Gly Ile Leu Gly Leu Gly Arg Thr Asn
Asp Ser Ser Tyr Asp Asn Pro 210 215
220Thr Phe Met Asp Ala Val Ala Glu Ser Asn Val Phe Lys Ser Asn Ile225
230 235 240Val Gly Phe Ala
Leu Ser Arg Ser Pro Ala Lys Asp Gly Thr Val Ser 245
250 255Phe Gly Thr Thr Asp Lys Asp Lys Tyr Thr
Gly Asp Ile Thr Tyr Thr 260 265
270Asp Thr Val Gly Ser Asp Ser Tyr Trp Arg Ile Pro Val Asp Asp Val
275 280 285Tyr Val Gly Gly Thr Ser Cys
Asp Phe Ser Asn Lys Ser Ala Ile Ile 290 295
300Asp Thr Gly Thr Ser Tyr Ala Met Leu Pro Ser Ser Asp Ser Lys
Thr305 310 315 320Leu His
Ser Leu Ile Pro Gly Ala Lys Ser Ser Gly Ser Tyr His Ile
325 330 335Ile Pro Cys Asn Thr Thr Thr
Lys Leu Gln Val Ala Phe Ser Gly Val 340 345
350Asn Tyr Thr Ile Ser Pro Lys Asp Tyr Val Gly Ala Thr Ser
Gly Ser 355 360 365Gly Cys Val Ser
Asn Ile Ile Ser Tyr Asp Leu Phe Gly Asp Asp Ile 370
375 380Trp Leu Leu Gly Asp Thr Phe Leu Lys Asn Val Tyr
Ala Val Phe Asp385 390 395
400Tyr Asp Glu Leu Arg Val Gly Phe Ala Glu Arg Ser Ser Asn Thr Thr
405 410 415Ser Ala Ser Asn Ser
Thr Ser Ser Gly Thr Ser Ser Thr Ser Gly Ser 420
425 430Thr Thr Thr Gly Ser Ser Thr Thr Thr Thr Ser Ser
Ala Ser Ser Ser 435 440 445Ser Ser
Ser Asp Ala Glu Ser Gly Ser Ser Met Thr Ile Pro Ala Pro 450
455 460Gln Tyr Phe Phe Ser Ala Leu Ala Ile Ala Ser
Phe Met Leu Trp Leu465 470 475
4801161099PRTAspergillus niger 116Met Leu Arg Gly Leu Arg Asp Val
Val Leu Leu Gln Phe Ala Ile Pro1 5 10
15Leu Phe Leu Leu Leu His Phe Arg Leu Ser Leu Arg Gly Val
Ile Thr 20 25 30Gly Phe Gly
Ser Lys Ser His Phe Gln Arg Pro Leu Ser Lys Met Ser 35
40 45Ser Thr Gln Lys Ser His Phe Lys Leu Leu Gln
Lys Phe Lys Pro Glu 50 55 60Tyr Ser
Pro Ser Glu Phe Ala Gln Tyr Glu Ser Glu Arg Thr Gly Met65
70 75 80Arg Val Val Val Ile Asp Gln
Lys Gly Pro Lys Val Thr Gly Tyr Phe 85 90
95Val Leu Ala Thr Glu Ile Leu Asp Asp Ser Gly Ala Pro
His Thr Leu 100 105 110Glu His
Leu Cys Phe Met Gly Ser Arg Asn Tyr Arg Tyr Lys Gly Phe 115
120 125Leu Asp Lys Leu Ala Thr Arg Val Tyr Ser
Ser Thr Asn Ala Trp Thr 130 135 140Ala
Thr Asp His Thr Ala Tyr Thr Leu Asp Thr Ala Gly Trp Glu Gly145
150 155 160Phe Ala Gln Ile Leu Pro
Val Tyr Leu Glu His Val Ile Ala Pro Thr 165
170 175Leu Thr Asp Glu Gly Cys Tyr Thr Glu Val His His
Ile Asp Gly Ala 180 185 190Gly
Asp Asp Ala Gly Val Val Tyr Ser Glu Met Gln Gly Val Gln Asn 195
200 205Asn Ser Ala Glu Leu Ile Asp Leu Thr
Ala Arg Arg Leu Thr Tyr Pro 210 215
220His Gly Val Gly Phe Arg Tyr Glu Thr Gly Gly Met Met Glu Gln Leu225
230 235 240Arg Val Leu Thr
Ala Asp Arg Ile Arg Ala Phe His Arg Glu Met Tyr 245
250 255Gln Pro Lys Asn Leu Cys Leu Ile Ile Thr
Gly Glu Val Asp His Gln 260 265
270Asn Met Leu Glu Thr Leu Asp Lys Phe Glu Asp Thr Ile Leu Asp Val
275 280 285Ile Pro Ser Pro Asp Ser Pro
Phe Lys Arg Pro Trp Val Asp Ser Lys 290 295
300Gln Ala Pro Pro Leu Glu Lys Ser Ile Val Gln Thr Val Glu Phe
Pro305 310 315 320Glu Glu
Asp Glu Ser Phe Gly Glu Ile Glu Ile Arg Phe Leu Gly Pro
325 330 335Asp Cys Thr Asp Pro Val Gln
Thr Gly Ala Val Asn Val Ala Leu Leu 340 345
350Tyr Leu Ala Gly Ser Ser Ala Ser Leu Leu Asp Asn Ile Leu
Val Glu 355 360 365Lys Glu Gln Leu
Ala Ser Ala Val Tyr Tyr Ala Thr Glu Asp His Pro 370
375 380Ser Ile Glu Ile Arg Phe Thr Leu Thr Ser Val Glu
Thr Glu Lys Leu385 390 395
400Ala Lys Val Glu Gln Arg Phe Phe Glu Val Leu Lys Asp Ala Met Glu
405 410 415Lys Asp Leu Asp Met
Arg Tyr Ile Lys Glu Cys Ile Asp Arg Gln Arg 420
425 430Arg Thr Trp Lys Phe Ser Thr Glu Ser Ser Ala Ser
Ser Phe Ala Glu 435 440 445Tyr Val
Ile Ser Asp Phe Leu Phe Gly Lys Arg Asp Gly Ser Thr Met 450
455 460Leu Asp Val Ala Thr Leu Gln Glu Tyr Asp Val
Leu Glu Lys Trp Ser465 470 475
480Glu Glu Gln Trp Arg Ser Phe Ile Lys Thr Trp Ile Ser Asp Ala Asn
485 490 495His Val Thr Ile
Leu Gly Val Pro Ser Val Lys Met Ser Asp Thr Leu 500
505 510Lys Lys Glu Glu Glu Ala Arg Val Ala Glu Gln
Lys Lys Arg Leu Gly 515 520 525Asp
Glu Gly Leu Lys Lys Leu Ala Asp Lys Leu Glu Lys Ala Lys Ala 530
535 540Glu Asn Asp Lys Glu Ile Pro Lys Glu Met
Leu Glu Arg Phe Gln Ile545 550 555
560Pro Gly Ile Glu Ser Ile His Phe Val Asp Thr Thr Thr Ala Arg
Ser 565 570 575Gly Ala Ala
Leu Asp Ala Gly Arg Pro Ser His Lys Ala Gln Lys Leu 580
585 590Val Asp Ala Asp Gly Ser Asp Leu Pro Leu
Phe Ile His Phe Glu His 595 600
605Ile Pro Ser Ser Phe Val Gln Leu Ser Leu Leu Ile Ser Ala Gln Ala 610
615 620Val Pro Val Gln Leu Arg Pro Leu
Leu Ser Val Tyr Thr Glu Ala Phe625 630
635 640Phe Asn Leu Pro Val Asn Arg Asn Gly Glu Thr Ile
Asn Phe Glu Gln 645 650
655Val Val Val Glu Leu Glu Arg Asp Thr Val Gly Tyr Ser Met Glu Gly
660 665 670Ala Arg Ser Leu Gly Asn
Ser Glu Met Leu Arg Ile Ser Phe Gln Val 675 680
685Glu Leu Glu Lys Tyr His Thr Ala Ile Ala Trp Ile Gln Glu
Leu Ser 690 695 700Trp Asn Ser Ile Phe
Asp Val Glu Arg Leu Arg Ala Ile Thr Ser Arg705 710
715 720Leu Leu Ser Asp Val Pro Asp Ser Lys Arg
Ser Gly Asp Asp Met Leu 725 730
735Ala Ala Val His Val Met Val His Tyr Ala Ala Glu Ser Ile Val Arg
740 745 750Ala Arg Ser Thr Leu
Val Lys Ala Arg Tyr Leu Lys Arg Ile Lys Lys 755
760 765Gln Leu Ala Glu Glu Pro Lys Ser Val Val Ala Arg
Met Glu Glu Ile 770 775 780Arg Asp Ala
Leu Phe Arg Phe Glu Asn Met Arg Val Leu Val Ile Ala785
790 795 800Asp Leu Glu Lys Leu Gln Asn
Pro Val Ser Ala Trp Lys Pro Phe Ala 805
810 815Glu Arg Leu Gly Ala Gly Ala Pro Leu Gln Pro Ile
Thr Thr Arg Arg 820 825 830Pro
Leu Leu Ser Glu Ala Gly Gln Lys Leu Gly Gly Lys Ser Tyr Val 835
840 845Val Pro Met Pro Thr Ile Asp Ser Ser
Phe Ala Tyr Ala Thr Ala Arg 850 855
860Gly Leu Asp Ser Tyr Asp Asp Pro Arg Leu Pro Ala Leu Met Val Ala865
870 875 880Ile Ala Tyr Met
Asn Ala Val Glu Gly Pro Leu Trp Val Ala Val Arg 885
890 895Gly Lys Gly Leu Ala Tyr Gly Thr Asn Phe
Ala Tyr Asn Ile Asp Thr 900 905
910Gly Phe Val Asn Phe Asp Val Tyr Arg Ser Pro Asn Ala His Lys Ala
915 920 925Phe Asp Ser Ser Lys Gln Ile
Val Glu Asp His Leu Ser Gly Ala Met 930 935
940Pro Phe Asp Pro Leu Met Leu Glu Gly Ser Ile Ser Ser Ile Val
Val945 950 955 960Ser Phe
Ala Asn Glu Gln Ser Thr Ile Gly Ser Ala Ala Ser Gly Ser
965 970 975Phe Ile Arg Gln Val Ile Arg
Arg Leu Pro Ser Asp Tyr Lys Glu Arg 980 985
990Val Leu Lys Gln Val Arg Ala Thr Ser Val Asp Asp Val Lys
Gly Ala 995 1000 1005Leu Lys Asp
Ile Ile Leu Pro Leu Phe Asn Pro Ser Thr Ala Asn 1010
1015 1020Ile Val Val Thr Cys Ala Thr Val Leu Glu Glu
Thr Ile Lys Glu 1025 1030 1035Gly Leu
Gln Ala Ser Gly Phe Thr Pro Ala Val Gln Pro Leu Lys 1040
1045 1050Glu Phe Glu Asp Asp Tyr Gly Leu Lys Val
Gly Asp Asp Glu Asp 1055 1060 1065Glu
Glu Ser Asp Asp Asp Asp Asp Glu Tyr Glu Thr Gly Ser Glu 1070
1075 1080Asp Glu Asp Asp Ser Asp Glu Asp Met
Glu Asp Asp Glu Asp Asp 1085 1090
1095Glu117726PRTAspergillus niger 117Met Gly Ala Leu Gln Trp Leu Ser Ile
Thr Ala Ala Ala Ala Ser Ala1 5 10
15Val Ser Ala Leu Thr Pro Glu Gln Met Ile Gly Ala Pro Arg Arg
Thr 20 25 30Glu Val Ile Pro
Asn Pro Ser Gly Asp Thr Gly Leu Phe Ser Thr Ser 35
40 45Gln Trp Ser Phe Asp Thr His Ser Glu Ser Thr Trp
Trp Ser Leu Ile 50 55 60Asp Leu Gln
Ser Gly Lys Thr Thr Thr Leu Thr Asp Asp Ser Asp Ile65 70
75 80Glu Glu Ile Ile Trp Leu Gly Ser
Asp Asn Ser Thr Leu Leu Tyr Ile 85 90
95Asn Ser Thr Asn Ala Gln Val Pro Gly Gly Val Glu Leu Trp
Ile Ala 100 105 110Asp Ser Ser
Asp Phe Ala Asn Ala Tyr Lys Ala Ala Ser Leu Ser Ala 115
120 125Gly Phe Leu Gly Ile Lys Ser Thr Val Thr Asp
Ser Gly Asp Val His 130 135 140Phe Ile
Leu Arg Gly Lys Ser Tyr Pro Asn Gly Thr Ala Tyr Asn Asp145
150 155 160Gln Leu Ala Glu Thr Tyr Pro
Ser Thr Ala Arg Ile Tyr Asp Ser Ile 165
170 175Phe Val Arg His Trp Asp Thr Tyr Leu Thr Thr Ala
Ser His Ala Val 180 185 190Phe
Ser Gly Thr Leu Gln Ser Ser Thr Ser Asp Asp Gly Asn Val Gln 195
200 205Tyr Thr Ser Ser Gly Gly Leu Thr Asn
Leu Val Asn Pro Val Lys Gly 210 215
220Ala Glu Ser Pro Phe Pro Pro Phe Gly Gly Asn Asp Asp Tyr Asp Leu225
230 235 240Ser Pro Asp Gly
Lys Trp Val Thr Phe Lys Ser Lys Ala Pro Glu Leu 245
250 255Pro Leu Ala Asn Asn Thr Ala Ala Tyr Val
Tyr Leu Val Pro His Asp 260 265
270Gly Ser Ala Thr Ala Phe Ala Val Asn Gly Pro Asp Ser Pro Ala Thr
275 280 285Pro Glu Gly Val Glu Gly Glu
Ser Asn Asn Pro Val Phe Ser Pro Asp 290 295
300Ser Asp Lys Ile Ala Tyr Phe Gln Met Ala Thr Asn Thr Tyr Glu
Ser305 310 315 320Asp Arg
Asn Val Leu Tyr Val Tyr Ser Ile Ala Asp Asp Thr Ile Thr
325 330 335Pro Leu Ala Lys Asp Trp Asp
Arg Ser Pro Ser Ser Val Thr Trp Val 340 345
350Asp Gly Asp Asn Leu Val Val Ala Ser Gln Asp Leu Gly Arg
Thr Arg 355 360 365Leu Phe Ala Ile
Pro Gly Asp Ala Gly Asp Asp Phe Lys Pro Thr Asn 370
375 380Phe Thr Asp Gly Gly Ser Val Ser Ala Gln Tyr Val
Leu Ser Asn Ser385 390 395
400Thr Leu Leu Val Thr Ser Ser Ala Phe Trp Thr Ser Trp Ser Val Tyr
405 410 415Thr Ala Ser Pro Asp
Glu Gly Val Ile Asn Thr Leu Ala Ser Ala Asn 420
425 430Glu Ile Asp Pro Glu Leu Ser Gly Leu Ser Ser Ser
Asp Phe Glu Glu 435 440 445Phe Tyr
Phe Asp Gly Asn Trp Thr Thr Leu Gln Gly Trp Ile Thr Tyr 450
455 460Pro Gln Asp Phe Asp Ser Ser Lys Lys Tyr Pro
Leu Ala Phe Leu Ile465 470 475
480His Gly Gly Pro Glu Asp Ala Trp Ala Asp Glu Trp Asn Leu Lys Trp
485 490 495His Ser Lys Val
Phe Ala Asp Gln Gly Tyr Val Val Val Gln Pro Asn 500
505 510Pro Thr Gly Ser Thr Gly Phe Gly Gln Gln Leu
Thr Asp Ala Ile Gln 515 520 525Leu
Asn Trp Thr Gly Ala Ala Tyr Asp Asp Leu Thr Lys Ala Trp Gln 530
535 540Tyr Val His Asp Thr Tyr Asp Phe Ile Asp
Thr Asp Asn Gly Val Ala545 550 555
560Ala Gly Pro Ser Phe Gly Ala Phe Met Ile Thr Trp Ile Gln Gly
Asp 565 570 575Asp Phe Gly
Arg Lys Phe Lys Ala Leu Val Ser His Asp Gly Pro Phe 580
585 590Ile Gly Asp Ala Trp Val Glu Thr Asp Glu
Leu Trp Phe Val Glu His 595 600
605Glu Phe Asn Gly Thr Phe Trp Gln Ala Arg Asp Ala Phe His Asn Thr 610
615 620Asp Pro Ser Gly Pro Ser Arg Val
Leu Ala Tyr Ser Thr Pro Gln Leu625 630
635 640Val Ile His Ser Asp Lys Asp Tyr Arg Ile Pro Val
Ala Asn Gly Ile 645 650
655Gly Leu Phe Asn Thr Leu Gln Glu Arg Gly Val Pro Ser Arg Phe Leu
660 665 670Asn Phe Pro Asp Glu Asp
His Trp Val Thr Gly Gln Glu Asn Ser Leu 675 680
685Val Trp Tyr Gln Gln Val Leu Gly Trp Ile Asn Arg Tyr Ser
Gly Val 690 695 700Gly Gly Ser Asn Pro
Asp Ala Ile Ala Leu Glu Asp Thr Val Asn Pro705 710
715 720Val Val Asp Leu Asn Pro
725118564PRTAspergillus niger 118Met Thr Arg Gln Thr Ser Leu Val Pro Arg
Leu Leu Thr Leu Ala Ser1 5 10
15Leu Ala Ala Leu Ser Gln Ala Glu Leu Gly Lys Ile Gln Trp Lys Gly
20 25 30Ser Cys Asn Leu Thr Thr
Tyr Pro Ala Leu Ile Cys Gly Thr Leu Asp 35 40
45Val Pro Tyr Asp Tyr Thr Glu Ser Asn Ser Ser Lys Thr Leu
Thr Leu 50 55 60Asp Ile Ala Lys Trp
Pro Ala Thr Lys Lys Pro Val Ser Glu Pro Ile65 70
75 80Ile Phe Asn Phe Gly Gly Pro Gly Val Asn
Ser Phe Glu Gly Leu Gly 85 90
95Leu Tyr Gly Glu Glu Phe Gln Ala Ile Leu Gly Gly His Asn Asp Leu
100 105 110Ile Ala Phe Asn Asn
Arg Gly Val Gly Asn Thr Ile Pro Phe Ser Cys 115
120 125Tyr Ser Asp Asp Ala Thr Arg Glu Leu Val Ala Leu
Gln Ala Pro Asn 130 135 140Asp Gly Arg
Ala Ser Ser Thr Ala Leu Gly Glu Ile Trp Ala Gln Asn145
150 155 160Ala Asn Ile Ala Gln Ala Cys
Tyr Ala Thr Asn Asn Gln Thr Gly Ser 165
170 175Leu Ile Gly Thr Ser Phe Ala Ala Arg Asp Ile Met
Gln Val Ala Asp 180 185 190Ala
Leu Ser Gly Lys Asp Ser Leu Val Asn Tyr Trp Gly Phe Ser Tyr 195
200 205Gly Thr Thr Ile Gly Ala Val Leu Ala
Ala Met Phe Pro Asp Arg Met 210 215
220Gly Asn Val Ala Leu Asp Gly Val Asp Asn Pro Arg Glu Ala Leu Tyr225
230 235 240Gly Tyr Asn Ala
Gln Ala Val Val Asp Val Asp Lys Val Phe Glu Gly 245
250 255Phe Cys Thr Gly Cys Met Ala Ala Pro Asp
Leu Cys Pro Ile Ala Lys 260 265
270Glu Tyr Thr Ser Ala Ala Asn Leu Glu Ala Ala Ile Tyr Leu Met Leu
275 280 285Glu Asn Leu Lys Tyr Asn Pro
Ile Ala Ile Pro Glu Thr Gly Gly Ile 290 295
300Val Thr Trp Ser Asp Val Lys Ser Thr Ile Phe Glu Ala Met Tyr
Leu305 310 315 320Pro Ser
Ser Trp Pro Leu Thr Ser Glu Leu Leu Tyr Tyr Val Gln Thr
325 330 335Arg Asn Thr Thr Ile Leu Gly
Asn Ser Glu Val Tyr Asp Thr Ile Lys 340 345
350Ser Tyr Gly Gln Ser Ala Ser Leu Thr Ser Ala Ser Asp Glu
Val Gly 355 360 365Thr Ala Ile Thr
Cys Ser Asp Lys His Arg Ser Ala Thr Ile Lys Glu 370
375 380Val Leu Pro Tyr Val Lys Ala Arg Gln Ala Leu Thr
Lys Ile Gly Ser385 390 395
400Asp Gly Ser Asp Gly Asp Met Arg Cys Ala Gln Trp Asn Pro Lys Met
405 410 415Phe Ala Lys Glu Arg
Tyr Ser Gly Asp Phe Glu Val Lys Thr Ala Asn 420
425 430Pro Val Leu Ile Leu Ser Asn Thr Tyr Asp Pro Ala
Thr Pro Leu Pro 435 440 445Ala Ala
Lys Asn Leu Thr Glu Thr Phe Glu Gly Ser Val Leu Leu Glu 450
455 460Gln Asn Gly Tyr Gly His Thr Thr Leu Ser Met
Pro Ser Leu Cys Thr465 470 475
480Ala Lys Ala Val Arg Ala Tyr Phe Thr Asn Gly Thr Leu Pro Ala Asp
485 490 495Gly Thr Ile Cys
Gln Val Asp Val Pro Leu Phe Thr Asn Leu Thr Tyr 500
505 510Lys Asp Val Trp Pro Lys Ser Phe Gln Arg Ser
Val Glu Ser Arg Asp 515 520 525Asp
Ala Thr Ile Leu Lys Ala Leu Met Ser Val Arg Asp Lys Met Ser 530
535 540Arg Arg Arg Met Cys Ile Tyr Leu Tyr Thr
Asn Ser Ala Ser Trp Arg545 550 555
560Pro Glu Leu Pro119526PRTAspergillus niger 119Met Tyr Tyr Ser
Leu Trp Val Ala Ala Leu Val Ala Ala Leu Pro Val1 5
10 15Ser Arg Ala Gln Phe Val Ala Pro Pro Thr
Asp Leu Ile Pro Thr Lys 20 25
30Gly Tyr Leu Asp Ile Pro Val Arg Tyr Lys Gln Val Pro Thr Gly Ile
35 40 45Cys Glu Thr Asp Pro Ser Val Lys
Ser Phe Ser Gly Tyr Val Asp Val 50 55
60Ala Glu His Glu His Ile Phe Phe Trp Phe Phe Glu Ala Arg Asn Gln65
70 75 80Asp Pro Thr Glu Ala
Pro Leu Thr Val Trp Ile Asn Gly Gly Met Ser 85
90 95Asp Pro Gly Pro Gly Ser Ser Ser Met Ile Gly
Leu Phe Gln Glu His 100 105
110Gly Pro Cys Gly Ile Asp Ala Asn Gly Ser Val Tyr Asn Asn Pro Tyr
115 120 125Ser Trp Asn Asn Ala Ser Asn
Met Leu Tyr Ile Asp Gln Pro Val Gln 130 135
140Thr Gly Phe Ser Tyr Ser Ile Pro Val Pro Gly Tyr Val Asp Ser
Ser145 150 155 160Thr Asp
Asn Gly Phe Met Gly Ala Phe Pro Gln Tyr Ser Arg Glu Thr
165 170 175Phe His Phe Thr Thr Glu Ser
Tyr Gly Gly His Tyr Gly Pro Val Phe 180 185
190Asn Glu Tyr Ile Glu Glu Gln Asn Ala His Leu Gln Pro Gly
Ala Lys 195 200 205Lys Ile Gln Leu
Gly Ser Val Met Ile Gly Asn Gly Trp Tyr Asp Pro 210
215 220Ile Ile Gln Tyr Gln Ala Tyr Tyr Asn Phe Thr Val
Tyr Pro Gly Asn225 230 235
240Thr Tyr Asp Tyr Leu Pro Phe Asn Lys Ser Ile Ser Ser Leu Met Tyr
245 250 255Asn Asn Leu Tyr Gly
Pro Gly Asn Cys Leu Asp Gln Leu Tyr Asp Cys 260
265 270Ala Ala Arg Gly Ile Asp Glu Ile Cys Ser Thr Ala
Asp Asp Phe Cys 275 280 285Ala Asn
Glu Val Glu Asn Val Tyr Asp Ile Tyr Ser Gly Arg Asp Glu 290
295 300Tyr Asp Phe Arg Glu Leu Thr Pro Asp Pro Phe
Pro Tyr Glu Phe Tyr305 310 315
320Val Asp Tyr Leu Asn Lys Ala Ser Val Gln Ala Ala Ile Gly Ala Tyr
325 330 335Ile Asn Tyr Thr
Glu Ser Asn Asn Ala Val Gly Leu Ala Phe Ser Ser 340
345 350Thr Gly Asp Asp Gly Arg Leu Met Asn Thr Ile
Gln Asp Val Gly Lys 355 360 365Leu
Leu Lys Gln Gly Val Thr Val Val Met Tyr Ala Gly Asp Ala Asp 370
375 380Tyr Asn Cys Asn Trp Leu Gly Gly Glu Ala
Val Ser Leu Gln Val Lys385 390 395
400Ala Ala Asn Phe Ser Ser Ala Gly Tyr Thr Asn Ile Val Thr Ser
Asp 405 410 415Gly Val Thr
His Gly Gln Val Arg Gln Ala Gly Gln Phe Ala Phe Val 420
425 430Arg Val Tyr Glu Ser Gly His Glu Val Pro
Phe Tyr Gln Pro Leu Leu 435 440
445Ala Leu Glu Met Phe Glu Arg Val Ile Gly Gly Lys Asp Val Ala Thr 450
455 460Gly Lys Ile Pro Ile Ser Ser Ser
Leu Gln Thr Val Gly Thr Pro Lys465 470
475 480Ser Tyr Tyr Arg Glu Gly Asn Ser Thr Ile Gln Trp
Glu Val Leu Asp 485 490
495Ser Leu Ala Thr Tyr Asn Thr Thr Thr Asn Ala Pro Asn Pro Val Ser
500 505 510Arg Arg Leu Lys Arg Met
Gly Pro Ala Leu Arg Phe Gln Met 515 520
5251201156PRTAspergillus niger 120Met Ser Cys Val Trp Leu His Ile
His Lys Arg Ser Leu Leu Ser Val1 5 10
15Ala Thr Asn Asn Ser Val Ala Arg Ala Ala Ala Ser Thr Ser
Ala Ala 20 25 30Pro Pro Pro
Pro Ser Ser Pro Pro Pro Gly Ser Asn Thr Tyr Ser Pro 35
40 45Leu Tyr Arg Pro Ile Thr Asn Pro Ile Gly Phe
Thr Leu Ser Pro Ala 50 55 60Arg Ser
Leu Val Ser Arg Asn Pro Lys Phe Pro Ala Tyr Arg Arg Ser65
70 75 80Ser Arg His Phe Ser Leu Cys
Pro Ala Ala Ala Thr Pro Gly Val Thr 85 90
95Thr Ser Ile Cys Pro Gly Gln Ala Pro Val Arg Ser Leu
Ser Ser Leu 100 105 110Ile Ile
His Ser Thr Arg Pro Arg Ala Ile Arg Ile Arg Thr Asp Gln 115
120 125Met Asp Leu Asn Gly Asp Ala Gly Ala Lys
Arg Lys Arg Ser Ser Ile 130 135 140Thr
Thr Pro Ala Glu Arg Pro Val Lys His Leu Arg Pro Glu Ser Ser145
150 155 160Ala Leu Thr Pro Gly Asp
Ser Thr Pro Ala Asn Gly Thr Val Tyr Asp 165
170 175Val Glu Asp Asp Glu Asp Ala Ser Arg Leu Leu Pro
Val Gly Pro Ala 180 185 190Gln
Ala Asp Ser Pro Glu Trp Gln Ala Thr Ile Glu Glu Val Val Lys 195
200 205Ser Val Val Ser Ile His Phe Cys Gln
Thr Cys Ser Phe Asp Thr Glu 210 215
220Leu Ser Met Ser Ser Gln Ala Thr Gly Phe Val Val Asp Ala Glu Asn225
230 235 240Gly Tyr Ile Leu
Thr Asn Arg His Val Val Cys Pro Gly Pro Phe Trp 245
250 255Gly Tyr Cys Ile Phe Asp Asn His Glu Glu
Cys Asp Val Arg Pro Val 260 265
270Tyr Arg Asp Pro Val His Asp Phe Gly Ile Leu Lys Phe Asp Pro Lys
275 280 285Ala Ile Arg Tyr Met Lys Leu
Arg Glu Leu Lys Leu Gln Pro Asp Ala 290 295
300Ala Lys Val Gly Ser Glu Ile Arg Val Val Gly Asn Asp Ala Gly
Glu305 310 315 320Lys Leu
Ser Ile Leu Ser Gly Val Ile Ser Arg Leu Asp Arg Asn Ala
325 330 335Pro Glu Tyr Gly Asp Gly Tyr
Ser Asp Phe Asn Thr Asn Tyr Ile Gln 340 345
350Ala Ala Ala Ala Ala Ser Gly Gly Ser Ser Gly Ser Pro Val
Val Asn 355 360 365Ile Asp Gly His
Ala Ile Ala Leu Gln Ala Gly Gly Arg Ala Asp Gly 370
375 380Ala Ala Thr Asp Tyr Phe Leu Pro Leu Asp Arg Pro
Leu Arg Ala Leu385 390 395
400Glu Cys Ile Arg Arg Gly Glu Pro Val Thr Arg Gly Thr Ile Gln Thr
405 410 415Gln Trp Ile Leu Lys
Pro Phe Asp Glu Cys Arg Arg Leu Gly Leu Thr 420
425 430Pro Glu Trp Glu Ala Thr Val Arg Lys Ala Ala Pro
Thr Glu Thr Ser 435 440 445Met Leu
Val Ala Glu Ile Ile Leu Pro Glu Gly Pro Ala Asp Gly Lys 450
455 460Leu Glu Glu Gly Asp Val Leu Leu Gln Val Asn
Gly Val Leu Leu Thr465 470 475
480Gln Phe Ile Arg Leu Asp Asp Ile Leu Asp Ser Ser Val Gly Gln Thr
485 490 495Val Arg Leu Leu
Val Gln Arg Gly Gly Gln Asn Val Glu Ile Glu Cys 500
505 510Gln Val Gly Asp Leu His Ala Ile Thr Pro Asp
Arg Phe Val Thr Val 515 520 525Ala
Gly Gly Thr Phe His Asn Leu Ser Tyr Gln Gln Ser Arg Leu Tyr 530
535 540Ala Ile Ala Thr Arg Gly Val Tyr Val Cys
Glu Ala Ala Gly Ser Phe545 550 555
560Lys Leu Glu Asn Thr Leu Ser Gly Trp Ile Ile Asp Ser Val Asp
Lys 565 570 575Arg Pro Thr
Arg Asn Leu Asp Glu Phe Val Glu Val Met Arg Thr Ile 580
585 590Pro Asp Arg Ser Arg Val Val Ile Ser Tyr
Arg His Ile Arg Asp Leu 595 600
605His Thr Arg Gly Thr Ser Ile Val Tyr Ile Asp Arg His Trp His Pro 610
615 620Lys Met Arg Leu Ala Val Arg Asn
Asp Asp Thr Gly Leu Trp Asp Phe625 630
635 640Ser Asp Leu Ala Asp Pro Ile Pro Ala Leu Pro Pro
Val Pro Arg Lys 645 650
655Ala Asp Phe Ile Gln Leu Asp Gly Val Ser Gln Pro Ala Ala Ala Asp
660 665 670Ile Val Arg Ser Phe Val
Arg Val Ser Cys Thr Met Pro Leu Lys Leu 675 680
685Asp Gly Tyr Pro Gln Ala Lys Lys Thr Gly Phe Gly Leu Val
Val Asp 690 695 700Ala Glu Lys Gly Leu
Val Val Val Ser Arg Ala Ile Val Pro Tyr Asp705 710
715 720Leu Cys Asp Ile Asn Val Thr Val Ala Asp
Ser Ile Ile Val Asn Ala 725 730
735Lys Val Val Phe Leu His Pro Leu Gln Asn Tyr Ser Ile Ile Gln Tyr
740 745 750Asp Pro Ser Leu Val
Gln Ala Pro Val Gln Ser Ala Lys Leu Ala Thr 755
760 765Asp Tyr Ile Lys Gln Gly Gln Asp Thr Ile Phe Val
Gly Phe Asn Gln 770 775 780Asn Phe Arg
Ile Val Val Ala Lys Thr Ala Val Thr Asp Ile Thr Thr785
790 795 800Val Ser Ile Pro Ala Asn Ala
Ser Ala Pro Arg Tyr Arg Ala Ile Asn 805
810 815Leu Asp Ala Ile Thr Val Asp Thr Gly Leu Ser Gly
Gln Cys Ser Asn 820 825 830Gly
Val Leu Ile Gly Glu Asp Gly Val Val Gln Ala Leu Trp Leu Asn 835
840 845Tyr Leu Gly Glu Arg Thr Ser Asn Ser
His Lys Asp Val Glu Tyr His 850 855
860Leu Gly Phe Ala Thr Pro Ser Leu Leu Pro Val Leu Ser Lys Val Gln865
870 875 880Gln Gly Glu Met
Pro Glu Leu Arg Ile Leu Asn Met Glu Ser Tyr Val 885
890 895Val Gln Met Ser Gln Ala Arg Ile Met Gly
Val Ser Glu Glu Trp Ile 900 905
910Glu Lys Val Thr Gln Ala Asn Pro Ser Arg His Gln Leu Phe Met Val
915 920 925Arg Lys Val Asp Cys Pro Pro
Pro Gly Phe Asn Ser Ala Ala Asp Thr 930 935
940Phe Glu Glu Gly Asp Ile Ile Leu Thr Leu Asp Gly Gln Leu Ile
Thr945 950 955 960Arg Val
Ser Glu Leu Asp Ile Met Tyr Glu Lys Asp Thr Leu Glu Ala
965 970 975Leu Ile Val Arg Asn Gly Gln
Glu Met Arg Ile Gln Val Pro Thr Val 980 985
990Pro Thr Glu Asp Leu Glu Thr Asp Arg Ala Val Val Phe Cys
Gly Ala 995 1000 1005Val Leu Gln
Lys Pro His His Ala Val Arg Gln Gln Ile Ser Lys 1010
1015 1020Leu His Ser Glu Val Tyr Val Ser Ala Arg Ser
Arg Gly Ser Pro 1025 1030 1035Ser Tyr
Gln Tyr Gly Leu Ala Pro Thr Asn Phe Ile Thr Ala Val 1040
1045 1050Asn Gly Val Pro Thr Pro Asn Leu Asp Arg
Phe Ser Glu Glu Val 1055 1060 1065Ser
Lys Ile Pro Asp Asn Thr Tyr Phe Arg Leu Arg Ala Val Thr 1070
1075 1080Phe Asp Asn Val Pro Trp Val Val Thr
Val Lys Lys Asn Asp His 1085 1090
1095Tyr Phe Pro Met Ser Glu Tyr Ile Lys Asp Gln Ser Gln Pro Ser
1100 1105 1110Gly Trp Arg Thr Val Ser
His Asp Lys Asp Lys Tyr Lys Asp Gly 1115 1120
1125Ile Ala Pro Asp Ala Ala Asn Leu Asn Pro Asp Ala Met Asp
Glu 1130 1135 1140Gly Phe Asp Gly Val
Ser Asp Ile Glu Pro Asp Leu Glu 1145 1150
1155121536PRTAspergillus niger 121Met Arg Val Leu Pro Ala Ala Met
Leu Val Gly Ala Ala Thr Ala Ala1 5 10
15Val Pro Pro Phe Gln Gln Val Leu Gly Gly Asn Gly Ala Lys
His Gly 20 25 30Ala Asp His
Ala Ala Glu Val Pro Ala Asp His Ser Ala Asp Gly Phe 35
40 45Ser Lys Pro Leu His Ala Phe Gln Glu Glu Leu
Lys Ser Leu Ser Asp 50 55 60Glu Ala
Arg Lys Leu Trp Asp Glu Val Ala Ser Phe Phe Pro Glu Ser65
70 75 80Met Asp Gln Asn Pro Leu Phe
Ser Leu Pro Lys Lys His Asn Arg Arg 85 90
95Pro Asp Ser His Trp Asp His Ile Val Asp Gly Lys Leu
Glu Ala Tyr 100 105 110Asp Leu
Arg Val Lys Lys Thr Asp Pro Gly Ser Leu Gly Ile Asp Pro 115
120 125Gly Val Lys Gln Tyr Thr Gly Tyr Leu Asp
Asp Asn Glu Asn Asp Lys 130 135 140His
Leu Phe Tyr Trp Phe Phe Glu Ser Arg Asn Asp Pro Glu Asn Asp145
150 155 160Pro Val Val Leu Trp Leu
Asn Gly Gly Pro Gly Cys Ser Ser Leu Thr 165
170 175Gly Leu Phe Met Glu Leu Gly Pro Ser Ser Ile Asn
Lys Lys Ile Gln 180 185 190Pro
Val Tyr Asn Asp Tyr Ala Trp Asn Ser Asn Ala Ser Val Ile Phe 195
200 205Leu Asp Gln Pro Val Asn Val Gly Tyr
Ser Tyr Ser Asn Ser Ala Val 210 215
220Ser Asp Thr Val Ala Ala Gly Lys Asp Val Tyr Ala Leu Leu Thr Leu225
230 235 240Phe Phe Lys Gln
Phe Pro Glu Tyr Ala Lys Gln Asp Phe His Ile Ala 245
250 255Gly Glu Ser Tyr Ala Gly His Tyr Ile Pro
Val Phe Ala Ser Glu Ile 260 265
270Leu Ser His Lys Lys Arg Asn Ile Asn Leu Gln Ser Val Leu Ile Gly
275 280 285Asn Gly Leu Thr Asp Gly Tyr
Thr Gln Tyr Glu Tyr Tyr Arg Pro Met 290 295
300Ala Cys Gly Asp Gly Gly Tyr Pro Ala Val Leu Asp Glu Ser Ser
Cys305 310 315 320Gln Ser
Met Asp Asn Ala Leu Pro Arg Cys Gln Ser Met Ile Glu Ser
325 330 335Cys Tyr Ser Ser Glu Ser Ala
Trp Val Cys Val Pro Ala Ser Ile Tyr 340 345
350Cys Asn Asn Ala Leu Leu Ala Pro Tyr Gln Arg Thr Gly Gln
Asn Val 355 360 365Tyr Asp Val Arg
Gly Lys Cys Glu Asp Ser Ser Asn Leu Cys Tyr Ser 370
375 380Ala Met Gly Tyr Val Ser Asp Tyr Leu Asn Lys Pro
Glu Val Ile Glu385 390 395
400Ala Val Gly Ala Glu Val Asn Gly Tyr Asp Ser Cys Asn Phe Asp Ile
405 410 415Asn Arg Asn Phe Leu
Phe His Gly Asp Trp Met Lys Pro Tyr His Arg 420
425 430Leu Val Pro Gly Leu Leu Glu Gln Ile Pro Val Leu
Ile Tyr Ala Gly 435 440 445Asp Ala
Asp Phe Ile Cys Asn Trp Leu Gly Asn Lys Ala Trp Thr Glu 450
455 460Ala Leu Glu Trp Pro Gly Gln Ala Glu Tyr Ala
Ser Ala Glu Leu Glu465 470 475
480Asp Leu Val Ile Val Asp Asn Glu His Thr Gly Lys Lys Ile Gly Gln
485 490 495Val Lys Ser His
Gly Asn Phe Thr Phe Met Arg Leu Tyr Gly Gly Gly 500
505 510His Met Val Pro Met Asp Gln Pro Glu Ser Ser
Leu Glu Phe Phe Asn 515 520 525Arg
Trp Leu Gly Gly Glu Trp Phe 530
535122279PRTAspergillus niger 122Met Lys Phe Thr Asn Tyr Leu Leu Thr Thr
Ala Thr Leu Ala Ser Ser1 5 10
15Val Leu Ala Ala Pro Ala Pro Arg Thr Gly Leu Glu Asp Arg Leu Arg
20 25 30Ala Arg Ser Leu Gln Arg
Gln Ser His Pro Leu Ala Pro Ile Pro Leu 35 40
45Asp Thr Ser Thr Lys Glu Asn Ser Arg Leu Leu Glu Ala Asp
Glu Asn 50 55 60Thr Thr His Val Thr
Tyr Ser Ser Asn Trp Ala Gly Ala Val Arg Glu65 70
75 80Gln Pro Pro Pro Gln Gly Thr Tyr Ser Ala
Val Ser Ala Thr Phe Arg 85 90
95Val Pro Glu Pro Thr Ala Gln Gly Gly Ser Gly Thr Gln Ala Gly Ser
100 105 110Ala Trp Val Gly Ile
Asp Gly Asp Thr Tyr Ser Asn Ala Ile Leu Gln 115
120 125Thr Gly Val Asp Phe Tyr Val Glu Asn Gly Gln Thr
Tyr Asn Asp Ala 130 135 140Trp Tyr Glu
Trp Tyr Pro Asp Tyr Ala Tyr Asp Phe Asp Leu Asp Val145
150 155 160Ser Thr Gly Asp Thr Ile Val
Ala Lys Val Glu Ala Ile Ser Pro Ser 165
170 175Gln Gly Val Ala Thr Ile Glu Asn Ile Ser Thr Gly
Lys Lys Ala Thr 180 185 190Gln
Thr Ile Arg Ala Pro Ala Ala Thr Ala Thr Leu Ala Gly Gln Asn 195
200 205Ala Asp Trp Ile Val Glu Asp Phe Gln
Ser Gly Asp Ser Met Val Asp 210 215
220Leu Ala Gly Phe Gly Glu Ile Ser Phe Trp Gly Val Gln Ala Gln Gly225
230 235 240Gly Gly Ser Thr
Trp Gly Val Asp Asp Ala Thr Ile Val Glu Leu Lys 245
250 255Gln Gly Asn Glu Val Leu Thr Asp Val Glu
Val Gln Ser Asp Ser Ala 260 265
270Phe Thr Val Lys Tyr Thr Ser 275123573PRTAspergillus niger
123Met Ile Tyr Val Asn Tyr Ile Leu Gly Leu Leu Ser Leu Leu His Thr1
5 10 15Ala Val Ala Thr Ala Pro
Asp Tyr Val Val Val Asp Gln Leu Asn Ser 20 25
30Ile Pro Asp Gly Trp Thr Lys Gly Ala Ala Pro Pro Pro
Phe Thr Pro 35 40 45Met Lys Phe
Trp Leu Ser Met His His Glu Tyr Lys Ala Asp Phe Glu 50
55 60Gln Lys Val Ile Asp Ile Ser Thr Pro Gly His Arg
Asp Tyr Gly Arg65 70 75
80His Met Lys Arg Asn Asp Val Met Ala Phe Met Arg Pro Ser Asp Gln
85 90 95Val Ser Lys Ile Ile Phe
Ser Trp Leu Glu Ser Glu His Val Pro Pro 100
105 110Asn Ala Ile Glu Asp Arg Gly Asp Trp Val Ala Phe
Thr Val Pro Leu 115 120 125Ala Gln
Ala Gln Ser Met Met Lys Thr Asp Phe Tyr Asn Phe His His 130
135 140Leu Glu Thr Asn Thr Thr Gln Ile Arg Thr Leu
Lys Tyr Ser Val Pro145 150 155
160Glu Gln Val Asp Ala His Leu Gln Met Ile Gln Pro Thr Thr Arg Phe
165 170 175Gly Arg Pro Lys
Thr Gln Thr Ser Leu Pro Ser Leu Met Pro Val Ser 180
185 190Val Asn Ile Asp Glu Ile Ser Glu Asp Cys Leu
Thr Gly Val Thr Pro 195 200 205Ile
Cys Leu Arg Gln Leu Tyr Gly Leu Pro Ser Thr Lys Ala Ser Pro 210
215 220Asp Ser Arg Asn Val Leu Gly Ile Ser Gly
Tyr Leu Asp Gln Tyr Ala225 230 235
240Arg Tyr Ser Asp Leu Asp Glu Phe Leu Ala Val Tyr Ser Pro Asn
Ser 245 250 255Val Asp Ala
Asp Phe Ser Val Val Ser Ile Asn Gly Gly Gln Asn Pro 260
265 270Gln Asn Ser Gln Glu Gly Ser Thr Glu Ala
Ser Leu Asp Ile Gln Tyr 275 280
285Ala Leu Ser Met Ala Phe Asp Ala Asn Ala Thr Phe Tyr Thr Thr Ala 290
295 300Gly Arg Ala Pro Ser Pro Tyr Leu
Glu Gln Leu Gln Tyr Leu Val Gly305 310
315 320Leu Pro Asp Glu Asp Leu Pro Ala Val Leu Ser Thr
Ser Tyr Gly Glu 325 330
335Asp Glu Gln Ser Leu Pro Glu Glu Tyr Thr Glu Ala Thr Cys Asn Leu
340 345 350Phe Ala Gln Leu Gly Ala
Arg Gly Val Ser Val Ile Phe Ser Ser Gly 355 360
365Asp Ser Gly Val Gly Gly Ser Cys Val Ser Asn Asp Gly Ser
Gln Arg 370 375 380Thr Arg Phe Gln Pro
Ile Phe Pro Ala Ser Cys Pro Phe Val Thr Ser385 390
395 400Val Gly Gly Thr Glu Gly Val Gly Pro Glu
Lys Ala Val Asp Phe Ser 405 410
415Ser Gly Gly Phe Ser Glu Arg Phe Ala Arg Pro Ser Tyr Gln Asn Ala
420 425 430Ser Val Glu Ala Tyr
Leu Ala Arg Leu Gly Asp Lys Trp Asp Gly Leu 435
440 445Tyr Asn Pro Asp Gly Arg Gly Ile Pro Asp Val Ser
Ala Gln Ala Ser 450 455 460Asn Tyr Val
Ile Arg Asp His Gly Gln Trp Leu Gln Thr Ala Gly Thr465
470 475 480Ser Ala Ala Ala Pro Val Phe
Ala Ala Val Ile Ser Arg Leu Asn Ala 485
490 495Ala Arg Leu Glu Gln Gly Lys Pro Thr Leu Gly Phe
Leu Asn Pro Trp 500 505 510Leu
Tyr Ser Leu Asp Gln Gln Gly Phe Thr Asp Ile Val Asp Gly Gly 515
520 525Ser Val Gly Cys Asp Gly Ser Asn Gly
Gly Ala Leu Val Pro Tyr Ala 530 535
540Ser Trp Asn Ala Thr Lys Gly Trp Asp Pro Val Thr Gly Leu Gly Thr545
550 555 560Pro Leu Tyr Gln
Thr Leu Glu Gln Leu Ala Gln Ser Ala 565
570124585PRTAspergillus niger 124Met Arg Ser Ser Gly Leu Tyr Thr Ala Leu
Leu Cys Ser Leu Ala Ala1 5 10
15Ser Thr Asn Ala Ile Val His Glu Lys Leu Ala Ala Val Pro Ser Gly
20 25 30Trp His His Val Glu Asp
Ala Gly Ser Asp His Gln Ile Ser Leu Ser 35 40
45Ile Ala Leu Ala Arg Lys Asn Leu Asp Gln Leu Glu Ser Lys
Leu Lys 50 55 60Asp Leu Ser Thr Pro
Gly Glu Ser Gln Tyr Gly Gln Trp Leu Asp Gln65 70
75 80Glu Asp Val Asp Thr Leu Phe Pro Val Ala
Ser Asp Lys Ala Val Ile 85 90
95Asn Trp Leu Arg Ser Ala Asn Ile Thr His Ile Ser Arg Gln Gly Ser
100 105 110Leu Val Asn Phe Ala
Thr Thr Val Asp Lys Val Asn Lys Leu Leu Asn 115
120 125Ala Thr Phe Ala Tyr Tyr Gln Ser Gly Ser Ser Gln
Arg Leu Arg Thr 130 135 140Thr Glu Tyr
Ser Ile Pro Asp Asp Leu Val Asp Ser Ile Asp Leu Ile145
150 155 160Ser Pro Thr Thr Phe Phe Gly
Lys Glu Lys Thr Thr Ala Gly Leu Asn 165
170 175Gln Arg Ala Gln Lys Ile Asp Thr His Val Ala Lys
Arg Ser Asn Ser 180 185 190Ser
Ser Cys Ala Asp Val Ile Thr Leu Ser Cys Leu Lys Glu Met Tyr 195
200 205Asn Phe Gly Asn Tyr Thr Pro Ser Ala
Ser Ser Gly Ser Lys Leu Gly 210 215
220Phe Gly Ser Phe Leu Asn Glu Ser Ala Ser Tyr Ser Asp Leu Ala Lys225
230 235 240Phe Glu Lys Leu
Phe Asn Leu Pro Ser Gln Ser Phe Ser Val Glu Leu 245
250 255Val Asn Gly Gly Val Asn Asp Gln Asn Gln
Ser Thr Ala Ser Leu Thr 260 265
270Glu Ala Asp Leu Asp Val Glu Leu Leu Val Gly Val Ala His Pro Leu
275 280 285Pro Val Thr Glu Phe Ile Thr
Ser Gly Glu Pro Ala Ala Asp Asn Glu 290 295
300Asn Glu Pro Tyr Leu Gln Tyr Tyr Glu Tyr Leu Leu Ser Lys Pro
Asn305 310 315 320Ser Ala
Leu Pro Gln Val Ile Ser Asn Ser Tyr Gly Asp Asp Glu Gln
325 330 335Thr Val Pro Glu Tyr Tyr Ala
Lys Arg Val Cys Asn Leu Ile Gly Leu 340 345
350Val Gly Leu Arg Gly Ile Ser Val Leu Glu Ser Ser Gly Asp
Glu Gly 355 360 365Ile Gly Ser Gly
Cys Arg Thr Thr Asp Gly Thr Asn Arg Thr Gln Phe 370
375 380Asn Pro Ile Phe Pro Ala Thr Cys Pro Tyr Val Thr
Ala Val Gly Gly385 390 395
400Thr Met Ser Tyr Ala Pro Glu Ile Ala Trp Glu Ala Ser Ser Gly Gly
405 410 415Phe Ser Asn Tyr Phe
Glu Arg Ala Trp Phe Gln Lys Glu Ala Val Gln 420
425 430Asn Tyr Leu Ala His His Ile Thr Asn Glu Thr Lys
Gln Tyr Tyr Ser 435 440 445Gln Phe
Ala Asn Phe Ser Gly Arg Gly Phe Pro Asp Val Ala Ala His 450
455 460Ser Phe Glu Pro Ser Tyr Glu Val Ile Phe Tyr
Gly Ala Arg Tyr Gly465 470 475
480Ser Gly Gly Thr Ser Ala Ala Cys Pro Leu Phe Ser Ala Leu Val Gly
485 490 495Met Leu Asn Asp
Ala Arg Leu Arg Ala Gly Lys Ser Thr Leu Gly Phe 500
505 510Leu Asn Pro Leu Leu Tyr Ser Lys Gly Tyr Arg
Ala Leu Thr Asp Val 515 520 525Thr
Gly Gly Gln Ser Ile Gly Cys Asn Gly Ile Asp Pro Gln Asn Asp 530
535 540Glu Thr Val Ala Gly Ala Gly Ile Ile Pro
Trp Ala His Trp Asn Ala545 550 555
560Thr Val Gly Trp Asp Pro Val Thr Gly Leu Gly Leu Pro Asp Phe
Glu 565 570 575Lys Leu Arg
Gln Leu Val Leu Ser Leu 580
585125265PRTAspergillus niger 125Met Lys Thr Thr Ala Leu Leu Thr Ala Gly
Leu Leu Ala Thr Thr Ala1 5 10
15Met Ala Ala Pro Leu Thr Ala Lys Arg Gln Ala Ala Arg Ala Lys Arg
20 25 30Ser Thr Asn Arg Gln Ser
Asn Pro Pro Phe Lys Pro Gly Thr Asn Glu 35 40
45Val Leu Ala Leu Asn Gly Thr Lys Asn Val Glu Tyr Ser Ser
Asn Trp 50 55 60Ala Gly Ala Val Leu
Ile Gly Thr Gly Tyr Thr Ala Val Thr Ala Glu65 70
75 80Phe Val Val Pro Thr Pro Ser Val Pro Ser
Gly Gly Ser Ser Arg Glu 85 90
95Glu Tyr Cys Ala Ser Ala Trp Val Gly Ile Asp Gly Asp Thr Cys Asp
100 105 110Thr Ala Ile Leu Gln
Thr Gly Val Asp Phe Cys Val Gln Gly Ser Glu 115
120 125Val Ser Phe Asp Ala Trp Tyr Glu Trp Tyr Pro Asp
Tyr Ala Tyr Asp 130 135 140Phe Ser Gly
Ile Ser Ile Ser Ala Gly Asp Thr Ile Lys Val Thr Val145
150 155 160Asp Ala Ser Ser Asp Thr Thr
Gly Thr Ala Thr Ile Glu Asn Val Ser 165
170 175Thr Gly Thr Thr Val Thr His Ser Phe Thr Gly Gly
Val Asp Gly Asp 180 185 190Leu
Cys Glu Tyr Asn Ala Glu Trp Ile Val Glu Asp Phe Glu Glu Asp 195
200 205Asp Ser Leu Val Pro Phe Ala Asp Phe
Gly Thr Val Thr Phe Thr Ser 210 215
220Cys Ser Ala Thr Lys Asp Gly Ser Ser Val Gly Pro Glu Asp Ala Thr225
230 235 240Ile Ile Asp Ile
Glu Gln Asn Glu Val Leu Thr Ser Val Ser Val Ser 245
250 255Ser Ser Glu Val Val Val Lys Tyr Val
260 265126580PRTAspergillus niger 126Met Val Ala Phe
Ser Arg Ile Ser Ala Gly Phe Ala Leu Ala Ala Pro1 5
10 15Ala Leu Ala Ser Val Val Leu Glu Thr Val
Lys Ser Val Pro Ser Asp 20 25
30Trp Lys Leu Val Glu Ala Ala Asp Thr Ser Ser Thr Ile Ser Leu Ser
35 40 45Val Ala Leu Ala Arg Gln Asn Leu
Asp Gln Leu Glu Glu Lys Leu Leu 50 55
60Ala Val Ser Thr Pro Gly Lys Asp Thr Tyr Gly Gln Phe Leu Asp Leu65
70 75 80Asp Asp Ile Asn Glu
Gln Phe Pro Leu Ala Asp Asp Ala Ala Val Val 85
90 95Ala Trp Leu Lys Lys Ala Gly Val Thr Gln Ile
His Lys Glu Gly Gly 100 105
110Leu Leu Asn Phe Ala Thr Thr Val Gly Thr Ala Asn Gln Leu Leu Asn
115 120 125Thr Thr Phe Ser Val Tyr Lys
Ser Gly Ser Thr Gln Lys Leu Arg Thr 130 135
140Thr Gln Tyr Ser Val Pro Asp Glu Leu Thr Gly Ser Ile Asp Leu
Ile145 150 155 160Ser Pro
Thr Val Phe Phe Gly Lys Ser Asn Ala Ala Arg Ser Ala Ala
165 170 175Val Arg Ala Ser Gln Thr Thr
Lys Glu Thr Ser Arg Lys Lys Ser Ser 180 185
190Asn Val Cys Glu Tyr Ile Thr Pro Asp Cys Leu Lys Glu Gln
Tyr Ser 195 200 205Ile Asp Tyr Thr
Pro Glu Ala Ser Ser Gly Ser Arg Val Gly Phe Gly 210
215 220Ser Phe Leu Asn Glu Ser Ala Leu Tyr Ser Asp Leu
Asp Leu Phe Thr225 230 235
240Gln Tyr Phe Asp Ile Pro Gln Gln Ser Phe Thr Val Glu Thr Ile Asn
245 250 255Gly Gly Ile Asn Asn
Gln Glu Asn Asp Pro Asp Gly Glu Ala Asp Leu 260
265 270Asp Val Gln Asn Ile Val Gly Ile Ser His Pro Leu
Pro Val Thr Glu 275 280 285Tyr Ile
Thr Gly Gly Ser Pro Pro Phe Ile Pro Asp Val Glu Thr Thr 290
295 300Thr Asp Glu Asn Glu Pro Tyr Leu Gln Tyr Tyr
Glu Tyr Leu Leu Ala305 310 315
320Lys Thr Asn Asp Glu Leu Pro Leu Val Ile Ser Asn Ser Tyr Gly Asp
325 330 335Asp Glu Asp Thr
Val Pro Ile Ala Tyr Ala Thr Arg Val Cys Asn Leu 340
345 350Ile Gly Leu Met Gly Thr Arg Gly Ile Ser Ile
Leu Glu Ser Ser Gly 355 360 365Asp
Ser Gly Val Gly Gly Ala Cys Met Ser Asn Asp Gly Thr Asp Lys 370
375 380Thr Glu Phe Thr Pro Met Phe Pro Gly Thr
Cys Pro Tyr Ile Thr Ala385 390 395
400Val Gly Gly Thr Gln Asp Val Pro Glu Val Ala Trp Val Asp Ser
Ser 405 410 415Gly Gly Phe
Ser Asn Tyr Phe Ser Gln Pro Ser Tyr Gln Ser Asp Gln 420
425 430Val Glu Thr Tyr Leu Asp Lys Tyr Ile Ser
Ala Ser Thr Lys Lys Tyr 435 440
445Tyr Glu Gln Tyr Thr Asn Phe Ser Gly Arg Ala Phe Pro Asp Val Ser 450
455 460Ala Phe Ala Gly Ser Pro Tyr Tyr
Glu Thr Tyr Ile Asp Gly Gln Leu465 470
475 480Gly Leu Val Ala Gly Thr Ser Gly Ala Ser Pro Val
Phe Ala Gly Ile 485 490
495Val Ala Leu Leu Asn Asp Ala Arg Leu Arg Ala Asn Lys Thr Ser Leu
500 505 510Gly Phe Leu Asn Pro Trp
Leu Tyr Ser Ser Gly Tyr Lys Ser Leu Asn 515 520
525Asp Ile Thr Ser Gly Glu Ala Val Gly Cys Gln Gly Asp Val
Glu Gly 530 535 540Ala Gly Val Ile Pro
Trp Ala Ser Trp Asn Ala Thr Thr Gly Trp Asp545 550
555 560Pro Ala Thr Gly Leu Gly Thr Pro Asn Phe
Ala Lys Leu Lys Glu Ala 565 570
575Val Leu Ala Leu 580127631PRTAspergillus niger 127Met
His Gly Leu Arg Leu Val Cys Ser Ile Gly Thr Leu Pro Leu Val1
5 10 15Ile Leu Ala Tyr Pro Ala Ala
Ser Leu His Thr Thr Ser Ala Ala Val 20 25
30Asp Leu Asp Ser Leu Arg Leu Thr Ser Asn Ser Glu Tyr Val
Asn Ser 35 40 45Val His Val Asp
Thr Asn Arg Ser Val Ala Val Ser Ala Glu Glu His 50 55
60Tyr Thr Asp Thr Ala Ala Arg Leu Val Gln Asn Ile Val
Pro Gly Ala65 70 75
80Ser Phe Arg Leu Ile Asp Asp His Phe Val Gly Asp Asn Gly Val Ala
85 90 95His Val Tyr Phe Arg Gln
Thr Leu His Gly Ile Asp Ile Asp Asn Ala 100
105 110Asp Phe Asn Val Asn Ile Gly Lys Asp Gly Leu Val
Leu Ser Phe Gly 115 120 125His Ser
Phe Phe Thr Gly Ala Leu Pro Ser Ser His Leu Asp Asn Thr 130
135 140Asn Val Leu Ser Pro Glu Ala Ala Leu Arg Gly
Ala Arg Asp Ala Ile145 150 155
160Gln Leu Pro Leu Thr Ile Asp Asn Val Ser Thr Glu Ala Ala Glu Gly
165 170 175Arg Asn Glu Tyr
Ile Phe Arg Glu Ala Val Gly Ala Val Ser Asp Pro 180
185 190Lys Ala Lys Leu Val Tyr Leu Val Lys Pro Glu
Gly Thr Leu Ala Leu 195 200 205Thr
Trp Arg Ile Glu Thr Asp Met Tyr Glu His Trp Leu Leu Thr Tyr 210
215 220Ile Asp Ala Glu Thr Thr Thr Val His Gly
Val Val Asp Tyr Val Ala225 230 235
240Asp Ala Thr Tyr Gln Val Tyr Pro Trp Gly Thr Asn Asp Pro Ala
Glu 245 250 255Gly His Arg
Thr Ile Val Thr Asp Pro Trp Asp Leu Ser Ala Ser Ala 260
265 270Tyr Thr Trp Ile Ser Asp Gly Arg Asp Asn
Tyr Thr Thr Thr Arg Gly 275 280
285Asn Asn Ala Ile Ala His Trp Asn Pro Thr Gly Gly Gly Ser Tyr Leu 290
295 300Tyr Asn Leu Arg Pro Ser Asp Pro
Asn Leu Asn Phe Gln Trp Pro Tyr305 310
315 320Ser Pro Asn Met Ser Pro Pro Arg Ser Tyr Ile Asn
Ala Ser Ile Val 325 330
335Gln Leu Phe Tyr Thr Ala Asn Ala Tyr His Asp Leu Leu Tyr Thr Leu
340 345 350Gly Phe Thr Glu Ser Ala
Gly Asn Phe Gln Trp Asn Asn Ser Ala His 355 360
365Gly Gly Arg Asp Lys Asp Tyr Val Ile Leu Asn Ala Gln Asp
Gly Ser 370 375 380Gly Phe Ser Asn Ala
Asn Phe Ala Thr Pro Pro Asp Gly Ile Pro Gly385 390
395 400Arg Met Arg Met Tyr Ile Trp Ile Glu Ser
Thr Pro Ser Arg Asp Gly 405 410
415Ser Phe Asp Ala Gly Ile Val Ile His Glu Tyr Thr His Gly Val Ser
420 425 430Asn Arg Leu Thr Gly
Gly Ser His Asn Ala Gly Cys Leu Ser Ala Leu 435
440 445Glu Ser Gly Gly Met Gly Glu Gly Trp Gly Asp Phe
Met Ala Thr Ala 450 455 460Ile Arg Ile
Lys Pro Asn Asp Thr Arg Thr Thr Ser Tyr Thr Met Gly465
470 475 480Ala Trp Ala Asp Asn Asp Lys
Cys Gly Val Arg Asp Tyr Pro Tyr Ser 485
490 495Thr Ser Phe Thr Glu Asn Pro Leu Asn Tyr Thr Ser
Val Asn Thr Met 500 505 510Asn
Gly Val His Ala Ile Gly Thr Val Trp Ala Thr Met Leu Tyr Glu 515
520 525Val Leu Trp Asn Leu Ile Asp Lys Tyr
Gly Lys Asn Asp Gly Ser Arg 530 535
540Pro Val Phe Arg Asn Gly Val Pro Thr Asp Gly Lys Tyr Leu Met Met545
550 555 560Lys Leu Val Val
Asp Gly Met Ala Leu Gln Pro Cys Asn Pro Asn Phe 565
570 575Val Gln Ala Arg Asp Ala Ile Leu Asp Ala
Asp Ile Val Leu Thr Gly 580 585
590Gly Lys Asn Arg Cys Glu Ile Trp Arg Gly Phe Ala Lys Arg Gly Leu
595 600 605Gly Gln Gly Ala Ala His Ser
Ser Leu Asn Trp Met Arg Arg Gly Ser 610 615
620Thr Leu Leu Pro Thr Gly Cys625
630128394PRTAspergillus niger 128Met Val Val Phe Ser Lys Thr Ala Ala Leu
Val Leu Gly Leu Ser Ser1 5 10
15Ala Val Ser Ala Ala Pro Ala Pro Thr Arg Lys Gly Phe Thr Ile Asn
20 25 30Gln Ile Ala Arg Pro Ala
Asn Lys Thr Arg Thr Ile Asn Leu Pro Gly 35 40
45Met Tyr Ala Arg Ser Leu Ala Lys Phe Gly Gly Thr Val Pro
Gln Ser 50 55 60Val Lys Glu Ala Ala
Ser Lys Gly Ser Ala Val Thr Thr Pro Gln Asn65 70
75 80Asn Asp Glu Glu Tyr Leu Thr Pro Val Thr
Val Gly Lys Ser Thr Leu 85 90
95His Leu Asp Phe Asp Thr Gly Ser Ala Asp Leu Trp Val Phe Ser Asp
100 105 110Glu Leu Pro Ser Ser
Glu Gln Thr Gly His Asp Leu Tyr Thr Pro Ser 115
120 125Ser Ser Ala Thr Lys Leu Ser Gly Tyr Thr Trp Asp
Ile Ser Tyr Gly 130 135 140Asp Gly Ser
Ser Ala Ser Gly Asp Val Tyr Arg Asp Thr Val Thr Val145
150 155 160Gly Gly Val Thr Thr Asn Lys
Gln Ala Val Glu Ala Ala Ser Lys Ile 165
170 175Ser Ser Glu Phe Val Gln Asn Thr Ala Asn Asp Gly
Leu Leu Gly Leu 180 185 190Ala
Phe Ser Ser Ile Asn Thr Val Gln Pro Lys Ala Gln Thr Thr Phe 195
200 205Phe Asp Thr Val Lys Ser Gln Leu Asp
Ser Pro Leu Phe Ala Val Gln 210 215
220Leu Lys His Asp Ala Pro Gly Val Tyr Asp Phe Gly Tyr Ile Asp Asp225
230 235 240Ser Lys Tyr Thr
Gly Ser Ile Thr Tyr Thr Asp Ala Asp Ser Ser Gln 245
250 255Gly Tyr Trp Gly Phe Ser Thr Asp Gly Tyr
Ser Ile Gly Asp Gly Ser 260 265
270Ser Ser Ser Ser Gly Phe Ser Ala Ile Ala Asp Thr Gly Thr Thr Leu
275 280 285Ile Leu Leu Asp Asp Glu Ile
Val Ser Ala Tyr Tyr Glu Gln Val Ser 290 295
300Gly Ala Gln Glu Ser Glu Glu Ala Gly Gly Tyr Val Phe Ser Cys
Ser305 310 315 320Thr Asn
Pro Pro Asp Phe Thr Val Val Ile Gly Asp Tyr Lys Ala Val
325 330 335Val Pro Gly Lys Tyr Ile Asn
Tyr Ala Pro Ile Ser Thr Gly Ser Ser 340 345
350Thr Cys Phe Gly Gly Ile Gln Ser Asn Ser Gly Leu Gly Leu
Ser Ile 355 360 365Leu Gly Asp Val
Phe Leu Lys Ser Gln Tyr Val Val Phe Asn Ser Glu 370
375 380Gly Pro Lys Leu Gly Phe Ala Ala Gln Ala385
390129398PRTAspergillus niger 129Met Lys Ser Ala Ser Leu Leu Thr
Ala Ser Val Leu Leu Gly Cys Ala1 5 10
15Ser Ala Glu Val His Lys Leu Lys Leu Asn Lys Val Pro Leu
Glu Glu 20 25 30Gln Leu Tyr
Thr His Asn Ile Asp Ala His Val Arg Ala Leu Gly Gln 35
40 45Lys Tyr Met Gly Ile Arg Pro Ser Ile His Lys
Glu Leu Val Glu Glu 50 55 60Asn Pro
Ile Asn Asp Met Ser Arg His Asp Val Leu Val Asp Asn Phe65
70 75 80Leu Asn Ala Gln Tyr Phe Ser
Glu Ile Glu Leu Gly Thr Pro Pro Gln 85 90
95Lys Phe Lys Val Val Leu Asp Thr Gly Ser Ser Asn Leu
Trp Val Pro 100 105 110Ser Ser
Glu Cys Ser Ser Ile Ala Cys Tyr Leu His Asn Lys Tyr Asp 115
120 125Ser Ser Ala Ser Ser Thr Tyr His Lys Asn
Gly Ser Glu Phe Ala Ile 130 135 140Lys
Tyr Gly Ser Gly Ser Leu Ser Gly Phe Ile Ser Gln Asp Thr Leu145
150 155 160Lys Ile Gly Asp Leu Lys
Val Lys Gly Gln Asp Phe Ala Glu Ala Thr 165
170 175Asn Glu Pro Gly Leu Ala Phe Ala Phe Gly Arg Phe
Asp Gly Ile Leu 180 185 190Gly
Leu Gly Tyr Asp Thr Ile Ser Val Asn Lys Ile Val Pro Pro Phe 195
200 205Tyr Asn Met Leu Asp Gln Gly Leu Leu
Asp Glu Pro Val Phe Ala Phe 210 215
220Tyr Leu Gly Asp Thr Asn Lys Glu Gly Asp Glu Ser Val Ala Thr Phe225
230 235 240Gly Gly Val Asp
Lys Asp His Tyr Thr Gly Glu Leu Ile Lys Ile Pro 245
250 255Leu Arg Arg Lys Ala Tyr Trp Glu Val Glu
Leu Asp Ala Ile Ala Leu 260 265
270Gly Asp Asp Val Ala Glu Met Glu Asn Thr Gly Val Ile Leu Asp Thr
275 280 285Gly Thr Ser Leu Ile Ala Leu
Pro Ala Asp Leu Ala Glu Met Ile Asn 290 295
300Ala Gln Ile Gly Ala Lys Lys Gly Trp Thr Gly Gln Tyr Thr Val
Asp305 310 315 320Cys Asp
Lys Arg Ser Ser Leu Pro Asp Val Thr Phe Thr Leu Ala Gly
325 330 335His Asn Phe Thr Ile Ser Ser
Tyr Asp Tyr Thr Leu Glu Val Gln Gly 340 345
350Ser Cys Val Ser Ala Phe Met Gly Met Asp Phe Pro Glu Pro
Val Gly 355 360 365Pro Leu Ala Ile
Leu Gly Asp Ala Phe Leu Arg Lys Trp Tyr Ser Val 370
375 380Tyr Asp Leu Gly Asn Ser Ala Val Gly Leu Ala Lys
Ala Lys385 390 395130393PRTAspergillus
niger 130Met Arg Lys Tyr Arg Phe His Pro Thr Lys Pro Gly Pro Tyr Thr Leu1
5 10 15Ser Ser Ser Ile
Gln Gln Thr Gly Arg Pro Tyr Thr Glu Lys Pro Ile 20
25 30Gly Gly Arg Ala His Ile Arg Gln Leu Val Arg
Lys Lys Ser Thr Thr 35 40 45Ser
Asp Glu Val Gly Glu Val Pro Ala Glu Asp Val Gln Asn Asp Ser 50
55 60Met Tyr Leu Ala Thr Val Gly Ile Gly Thr
Pro Ala Gln Asn Leu Lys65 70 75
80Leu Asp Phe Asp Thr Gly Ser Ala Asp Leu Trp Val Trp Ser Asn
Lys 85 90 95Leu Pro Ser
Thr Leu Leu Ser Glu Asn Lys Thr His Ala Ile Phe Asp 100
105 110Ser Ser Lys Ser Ser Thr Phe Lys Thr Leu
Glu Gly Glu Ser Trp Gln 115 120
125Ile Ser Tyr Gly Asp Gly Ser Ser Ala Ser Gly Ser Val Gly Thr Asp 130
135 140Asp Val Asn Ile Gly Gly Val Val
Val Lys Asn Gln Ala Val Glu Leu145 150
155 160Ala Glu Lys Met Ser Ser Thr Phe Ala Gln Gly Glu
Gly Asp Gly Leu 165 170
175Leu Gly Leu Ala Phe Ser Asn Ile Asn Thr Val Gln Pro Lys Ser Val
180 185 190Lys Thr Pro Val Glu Asn
Met Ile Leu Gln Asp Asp Ile Pro Lys Ser 195 200
205Ala Glu Leu Phe Thr Ala Lys Leu Asp Thr Trp Arg Asp Thr
Asp Asp 210 215 220Glu Ser Phe Tyr Thr
Phe Gly Phe Ile Asp Gln Asp Leu Val Lys Thr225 230
235 240Ala Gly Glu Glu Val Tyr Tyr Thr Pro Val
Asp Asn Ser Gln Gly Phe 245 250
255Trp Leu Phe Asn Ser Thr Ser Ala Thr Val Asn Gly Lys Thr Ile Asn
260 265 270Arg Ser Gly Asn Thr
Ala Ile Ala Asp Thr Gly Thr Thr Leu Ala Leu 275
280 285Val Asp Asp Asp Thr Cys Glu Ala Ile Tyr Ser Ala
Ile Asp Gly Ala 290 295 300Tyr Tyr Asp
Gln Glu Val Gln Gly Trp Ile Tyr Pro Thr Asp Thr Ala305
310 315 320Gln Asp Lys Leu Pro Thr Val
Ser Phe Ala Val Gly Glu Lys Gln Phe 325
330 335Val Val Gln Lys Glu Asp Leu Ala Phe Ser Glu Ala
Lys Thr Gly Tyr 340 345 350Val
Tyr Gly Gly Ile Gln Ser Arg Gly Asp Met Thr Met Asp Ile Leu 355
360 365Gly Asp Thr Phe Leu Lys Ser Ile Tyr
Ala Val Ser Ala Leu Leu Leu 370 375
380Ala Leu Arg Gly Asp Ile Glu Ala His385
390131282PRTAspergillus niger 131Met Lys Phe Ser Thr Ile Leu Thr Gly Ser
Leu Phe Ala Thr Ala Ala1 5 10
15Leu Ala Ala Pro Leu Thr Glu Lys Arg Arg Ala Arg Lys Glu Ala Arg
20 25 30Ala Ala Gly Lys Arg His
Ser Asn Pro Pro Tyr Ile Pro Gly Ser Asp 35 40
45Lys Glu Ile Leu Lys Leu Asn Gly Thr Ser Asn Glu Asp Tyr
Ser Ser 50 55 60Asn Trp Ala Gly Ala
Val Leu Ile Gly Asp Gly Tyr Thr Lys Val Thr65 70
75 80Gly Glu Phe Thr Val Pro Ser Val Ser Ala
Gly Ser Ser Ser Ser Ser 85 90
95Gly Tyr Gly Gly Gly Tyr Gly Tyr Tyr Lys Asn Lys Arg Gln Ser Glu
100 105 110Glu Tyr Cys Ala Ser
Ala Trp Val Gly Ile Asp Gly Asp Thr Cys Glu 115
120 125Thr Ala Ile Leu Gln Thr Gly Val Asp Phe Cys Tyr
Glu Asp Gly Gln 130 135 140Thr Ser Tyr
Asp Ala Trp Tyr Glu Trp Tyr Pro Asp Tyr Ala Tyr Asp145
150 155 160Phe Asn Asp Ile Thr Ile Ser
Glu Gly Asp Thr Ile Lys Val Thr Val 165
170 175Glu Ala Thr Ser Lys Ser Ser Gly Ser Ala Thr Val
Glu Asn Leu Thr 180 185 190Thr
Gly Gln Ser Val Thr His Thr Phe Ser Gly Asn Val Glu Gly Asp 195
200 205Leu Cys Glu Thr Asn Ala Glu Trp Ile
Val Glu Asp Phe Glu Ser Gly 210 215
220Asp Ser Leu Val Ala Phe Ala Asp Phe Gly Ser Val Thr Phe Thr Asn225
230 235 240Ala Glu Ala Thr
Ser Asp Gly Ser Thr Val Gly Pro Ser Asp Ala Thr 245
250 255Val Met Asp Ile Glu Gln Asp Gly Thr Val
Leu Thr Glu Thr Ser Val 260 265
270Ser Gly Asp Ser Val Thr Val Thr Tyr Val 275
280132273PRTAspergillus niger 132Met Gly Asp Tyr Gly Pro Gly Val Ser Ser
Leu Thr Ala Gln Leu Pro1 5 10
15Gly Asn Pro Pro Val Ser Glu Thr Asp Gln Asp Glu Ile Ser Val Leu
20 25 30Val Thr Gly Phe Gly Pro
Phe Lys Ser Asn Leu Val Asn Ala Ser Tyr 35 40
45Leu Ile Ala Ser Ser Leu Pro Pro Ser Phe Thr Phe Ser Pro
Ala Ser 50 55 60Ser Asp Gly Ser Asp
Ala Val Pro Arg Arg Val Ser Ile Asn Val His65 70
75 80Pro Ser Pro Ile Pro Val Ala Tyr Ser Ser
Val Arg Thr Thr Leu Pro 85 90
95Val Ile Leu Asp Asp Tyr Ala Lys Thr His Gly Gly Arg Arg Pro Asp
100 105 110Ile Val Ile His Ile
Gly Ile Ala Ala Met Arg Asn Tyr Tyr Ser Val 115
120 125Glu Thr Gln Ala His Arg Asp Gly Tyr Leu Met Ser
Asp Ile Lys Gly 130 135 140Arg Ser Gly
Tyr Glu Asp Gly Glu Lys Leu Trp Arg Glu Leu Asp Leu145
150 155 160Pro Leu Val Leu Arg Ala Gly
Pro Ser Glu Gly His Ala Ser Glu Lys 165
170 175Lys His Leu Ser Pro Arg Pro Pro Asp Glu Asp Phe
Leu Ala Ala Trp 180 185 190Lys
Thr Phe Cys Pro Pro Glu Thr Asp Ala Arg Ile Ser Thr Asp Ala 195
200 205Gly Arg Tyr Leu Cys Glu Phe Ile Leu
Tyr Thr Ser Leu Ala Leu Ala 210 215
220Tyr Gln Ala Gly Glu Asp Arg Asn Val Thr Phe Phe His Val Pro Ala225
230 235 240Ser Cys Leu Asp
Glu Asp Ile Glu Thr Gly Lys Glu Val Ala Val Ala 245
250 255Leu Ile Lys Ala Leu Val Thr Ser Trp Ser
Glu Gln Gln His Ser Val 260 265
270Pro133542PRTAspergillus niger 133Met Gly Ser Arg Gln Gly Lys Ala Pro
Phe Gly Trp Gly Thr Gln Ser1 5 10
15Leu Ala His Phe Gly Ile Asn Pro Asp Leu Gly Leu His Asn Gln
Gln 20 25 30Asn Leu Asn Ser
Leu Ile Ser His Ser Ala Met Ala Thr Ala Leu Glu 35
40 45Thr Glu Tyr Ala Thr Ile Pro Ile Asp His Asn Asn
Ala Ser Ala Gly 50 55 60Thr Tyr Gln
Asn Arg Phe Trp Val Ser Asp Glu Phe Tyr Gln Pro Gly65 70
75 80Asn Pro Ile Phe Val Tyr Asp Thr
Gly Glu Ser Asp Gly Gly Ser Ile 85 90
95Ala Gln Ser Tyr Leu Thr Ser Thr Leu Ser Phe Phe Arg Glu
Phe Leu 100 105 110Ile Glu Phe
Asn Ala Met Gly Ile Ala Trp Glu His Arg Tyr Tyr Gly 115
120 125Asn Ser Thr Pro Ala Pro Val Ser Tyr Glu Thr
Pro Pro Glu Ala Trp 130 135 140Gln Tyr
Leu Thr Thr Lys Gln Ala Leu Ala Asp Leu Pro Tyr Phe Ala145
150 155 160Ser Asn Phe Ser Arg Glu Lys
Tyr Pro Asp Met Asp Leu Thr Pro Gln 165
170 175Gly Thr Pro Trp Ile Met Val Gly Gly Ser Tyr Ala
Gly Ile Arg Ala 180 185 190Ala
Leu Thr Arg Lys Glu Tyr Pro Glu Thr Ile Phe Ala Ala Phe Ser 195
200 205Ser Ser Ser Pro Val Glu Ala Gln Val
Asn Met Ser Ala Tyr Tyr Asp 210 215
220Gln Val Tyr Arg Gly Met Val Ala Ser Gly Trp Thr Asn Cys Ser Ala225
230 235 240Asp Ile His Ala
Ala Leu Glu Tyr Ile Asp Asp Gln Leu Ser Asp Glu 245
250 255Asp Thr Ala Thr Ser Val Lys Gln Leu Phe
Phe Gly Ser Gly Ala Glu 260 265
270Thr Asn Ser Asn Gly Asp Phe Thr Ala Ala Leu Thr Ala Ile Tyr Gly
275 280 285Tyr Phe Gln Ser Tyr Gly Met
Ala Gly Gly Ile Gly Gly Leu Gly Ala 290 295
300Phe Cys Glu Tyr Leu Glu Ile Asp Pro Lys Thr Asn Gly Thr Thr
Gly305 310 315 320Pro Asp
Gly Leu Ala Pro Thr Tyr Gly Gly Gln Tyr Val Ala Glu Arg
325 330 335Trp Ala Ala Trp Pro Thr Phe
Leu Glu Leu Val Asn Leu Asn Met Gly 340 345
350Thr Asn Cys Gly Pro Gln Asp Ala Ser Gln Pro Ile Asp Cys
Asp Phe 355 360 365Ser Lys Pro Tyr
Gly Asp Pro Ser Ala Ile Thr Trp Thr Trp Gln Tyr 370
375 380Cys Ser Glu Trp Gly Phe Phe Gln Ala Asn Asn Asp
Gly Pro His Ser385 390 395
400Leu Ala Ser Arg Tyr Gln Ser Val Glu Tyr Gln Gln Glu Val Cys Asn
405 410 415Arg Gln Phe Pro Asp
Ala Val Asp Lys Gly Leu Leu Pro Pro Ser Pro 420
425 430Arg Ala Asp Asp Val Asn Gln Glu Phe Gly Gly Trp
Thr Ile Arg Pro 435 440 445Ser Asn
Val Tyr Phe Ser Gly Gly Glu Phe Asp Pro Trp Arg Ser Leu 450
455 460Ser Ile Leu Ser Thr Glu Asp Phe Ala Pro Gln
Gly Val Glu Phe Thr465 470 475
480Ser Ala Ile Pro Ala Cys Gly Val Gln Thr Asn Glu Asp Thr Val Phe
485 490 495Gly Tyr Val Met
Gln Asn Ser Glu His Cys Phe Asp Phe Gln Ala Thr 500
505 510Pro Thr Val Gly Lys Leu Ser Arg Gly Ile Phe
Thr Ser Ala Leu Leu 515 520 525Gln
Trp Leu Glu Cys Phe Gly Gln Asn Ser Ser Gln Ser Arg 530
535 540134391PRTAspergillus niger 134Met Lys Leu Ser Ile
Ala Leu Ala Leu Gly Ala Thr Ala Ser Thr Gly1 5
10 15Val Leu Ala Ala Val Val Pro Gln Gln Glu Pro
Leu Ile Thr Pro Gln 20 25
30Asp Pro Pro Thr His His His Gln Glu Lys Phe Leu Ile Glu Leu Ala
35 40 45Pro Tyr Gln Thr Arg Trp Val Thr
Glu Glu Glu Lys Trp Asp Leu Lys 50 55
60Leu Asp Gly Val Asn Phe Ile Asp Ile Thr Glu Glu Arg Asn Thr Gly65
70 75 80Phe Tyr Pro Thr Leu
His Ala Gly Ser Tyr Val His Tyr Pro Pro Thr 85
90 95Met Lys His Ala Glu Lys Val Val Pro Leu Leu
Arg Gly Leu Ser Lys 100 105
110Asp Asn Met Glu Gln Asn Leu Asn Lys Phe Thr Ser Phe His Thr Arg
115 120 125Tyr Tyr Arg Ser Ser Thr Gly
Ile Glu Ser Ala Lys Trp Leu Tyr Ser 130 135
140Arg Val Ser Asp Val Ile Glu Gln Ser Gly Ala Ala Glu Tyr Gly
Ala145 150 155 160Thr Val
Glu Gln Phe Ala His Ser Trp Gly Gln Phe Ser Ile Ile Ala
165 170 175Arg Ile Pro Gly Gln Thr Asn
Lys Thr Val Val Leu Gly Ala His Gln 180 185
190Asp Ser Ile Asn Leu Phe Leu Pro Ser Ile Leu Ala Ala Pro
Gly Ala 195 200 205Asp Asp Asp Gly
Ser Gly Thr Val Thr Ile Leu Glu Ala Leu Arg Gly 210
215 220Leu Leu Gln Ser Asp Ala Ile Val Arg Gly Asn Ala
Ser Asn Thr Ile225 230 235
240Glu Phe His Trp Tyr Ser Ala Glu Glu Gly Gly Met Leu Gly Ser Gln
245 250 255Ala Ile Phe Ser Gln
Tyr Lys Arg Asp Lys Arg Asp Ile Lys Ala Met 260
265 270Leu Gln Gln Asp Met Thr Gly Tyr Thr Gln Gly Ala
Leu Asp Ala Gly 275 280 285Arg Gln
Glu Ala Ile Gly Ile Met Val Asp Tyr Val Asp Glu Gly Leu 290
295 300Thr Gln Phe Leu Lys Asp Val Thr Thr Glu Tyr
Cys Gly Ile Gly Tyr305 310 315
320Ile Glu Thr Arg Cys Gly Tyr Ala Cys Ser Asp His Thr Ser Ala Ser
325 330 335Lys Tyr Gly Tyr
Pro Ala Ala Met Ala Thr Glu Ser Glu Met Glu Asn 340
345 350Ser Asn Lys Arg Ile His Thr Thr Asp Asp Ser
Ile Arg Tyr Leu Ser 355 360 365Phe
Asp His Met Leu Glu His Ala Arg Leu Thr Leu Gly Phe Ala Tyr 370
375 380Glu Leu Ala Phe Ala Gln Phe385
390135442PRTAspergillus niger 135Met Arg Thr Thr Thr Ser Phe Ala Arg
Leu Ala Leu Ala Val Ala Ser1 5 10
15Val Gly Ile Val Phe Ala Ser Pro Thr Lys Asn Asn Asp Gly Lys
Leu 20 25 30Val Tyr Gly Ser
Pro Glu Ser Val Gly Met Ile Ser Ala Pro Leu His 35
40 45Gln Met Val Gln Asn Val Ser Ala Tyr Thr His Ala
Ala Asn Tyr Ser 50 55 60Lys Phe Ser
Tyr Asp Lys Val His Pro Ile Glu Pro Gly Ser Val Thr65 70
75 80Leu Val Ala Leu Asp Gly Val Ile
Val Ser Glu Phe Ala Leu Gly Lys 85 90
95Arg Asn Leu Tyr Ala Asp Val Asn Gly Thr Asn Leu Pro Arg
Tyr Leu 100 105 110Gln Glu Asp
Thr Thr Leu Asp Thr Val Tyr Asp Met Ala Ser Leu Thr 115
120 125Lys Leu Phe Thr Thr Val Ala Ala Leu Arg Glu
Leu Asp Ala Gly Arg 130 135 140Ile Ala
Leu Asn Val Thr Val Ala Thr Tyr Ile Pro Asp Phe Ala Thr145
150 155 160Asn Gly Lys Glu Asn Ile Thr
Ile Leu Glu Leu Phe Thr His Thr Ser 165
170 175Gly Phe Ala Ser Asp Pro Ser Pro Pro Leu Phe Ser
Ala Tyr Tyr Thr 180 185 190Thr
Tyr Asp Glu Arg Ile Lys Ala Ile Leu Thr Gln Lys Ile Ile Asn 195
200 205Thr Pro Gly Ser Thr Tyr Leu Tyr Leu
Asp Leu Asn Phe Met Ser Leu 210 215
220Gly Leu Val Ile Glu Thr Val Thr Gly Arg Ala Leu Asp Asp Leu Ile225
230 235 240Tyr Asp Phe Thr
Arg Pro Leu Glu Met Thr Ser Thr Phe Phe Asn Arg 245
250 255Gly Asn Ile Glu Gly Ser Thr Pro Gln Ser
Pro Asn Tyr Asp Arg Thr 260 265
270Ala Val Gln Glu Phe Gln Ile Ala Ala Leu Gly Pro Ser Glu Pro Gln
275 280 285Arg Pro Gln Pro Val Arg Gly
Thr Val His Asp Glu Asn Ala Trp Ser 290 295
300Leu Asp Gly Val Ser Gly His Ala Gly Leu Phe Ser Thr Val Arg
Asp305 310 315 320Thr Ala
Thr Phe Cys Gln Met Ile Leu Asn Asn Gly Thr Tyr Ala Gly
325 330 335Gln Arg Ile Leu Ser Arg Thr
Ala Val Asp Met Ile Phe Thr Asn Phe 340 345
350Asn Ala Arg Phe Pro Gly Asp Ala Arg Ser Leu Gly Phe Glu
Leu Asp 355 360 365Gln Tyr Ser Thr
Ala Gly Pro Met Ala Ser Leu Gln Thr Ala Ser His 370
375 380Thr Gly Phe Thr Gly Thr Thr Leu Val Met Asp Arg
Thr Tyr Asn Ala385 390 395
400Phe Trp Leu His Phe Ser Asn Arg Val His Pro Ser Arg Ala Trp Ser
405 410 415Ser Asn Thr Ile Val
Arg Glu Ala Ile Gly Tyr Trp Val Gly Lys Ser 420
425 430Leu Gly Leu Asp Val Ala Phe Ala Leu Leu
435 440136612PRTAspergillus niger 136Met Ala Ser Trp Leu
Leu Ser Thr Leu Leu Phe Leu Ser Pro Ser Leu1 5
10 15Val Ser Ala Lys Ser Ala Ala Asp Tyr Tyr Val
His Ser Leu Pro Gly 20 25
30Ala Pro Glu Gly Pro Leu Leu Lys Met His Ala Gly His Ile Glu Val
35 40 45Asp Pro Gln Asn Asn Gly Asn Leu
Phe Phe Trp His Tyr Gln Asn Arg 50 55
60His Ile Ala Asn Arg Gln Arg Thr Val Ile Trp Leu Asn Gly Gly Pro65
70 75 80Gly Cys Ser Ser Met
Asp Gly Ala Leu Met Glu Val Gly Pro Tyr Arg 85
90 95Leu Lys Asp Asn Glu Thr Leu Thr Tyr Asn Glu
Gly Ser Trp Asp Glu 100 105
110Phe Ala Asn Leu Leu Phe Val Asp Gln Pro Val Gly Thr Gly Phe Ser
115 120 125Tyr Val Asn Thr Asp Ser Tyr
Leu His Glu Leu Asp Glu Met Ser Ala 130 135
140Gln Phe Ile Val Phe Leu Glu Glu Trp Phe Arg Leu Phe Pro Glu
Tyr145 150 155 160Glu Arg
Asp Asp Ile Tyr Ile Ala Gly Glu Ser Tyr Ala Gly Gln His
165 170 175Ile Pro Tyr Ile Ala Lys Ala
Ile Gln Glu Arg Asn Lys Asn Val Gln 180 185
190Gly Lys Thr Ile Ala Ser Trp Asn Leu Lys Gly Leu Leu Ile
Gly Asn 195 200 205Gly Trp Ile Ser
Pro Asn Glu Gln Tyr Met Ser Tyr Leu Pro Tyr Ala 210
215 220Tyr Glu Glu Gly Leu Ile Lys Glu Gly Ser Arg Thr
Ala Lys Glu Leu225 230 235
240Glu Val Leu Gln Ser Val Cys Lys Ser Arg Leu Glu Thr Gly Lys Asn
245 250 255Lys Val His Leu Asn
Asp Cys Glu Lys Val Met Asn Ala Leu Leu Asp 260
265 270Lys Thr Val Glu Asp Asn Lys Cys Leu Asn Met Tyr
Asp Ile Arg Leu 275 280 285Arg Asp
Thr Thr Asp Ala Cys Gly Met Asn Trp Pro Thr Asp Leu Glu 290
295 300Asp Val Lys Pro Tyr Leu Gln Arg Glu Asp Val
Val Lys Ala Leu Asn305 310 315
320Ile Asn Pro Glu Lys Lys Ser Gly Trp Val Glu Cys Ser Gly Ala Val
325 330 335Ser Ser Ala Phe
Asn Pro Gln Lys Ser Pro Pro Ser Val Gln Leu Leu 340
345 350Pro Gly Leu Leu Glu Ser Gly Leu Gln Ile Leu
Leu Phe Ser Gly Asp 355 360 365Lys
Asp Leu Ile Cys Asn His Val Gly Thr Glu Gln Leu Ile Asn Asn 370
375 380Met Lys Trp Asn Gly Gly Thr Gly Phe Glu
Thr Ser Pro Gly Val Trp385 390 395
400Ala Pro Arg His Asp Trp Ser Phe Glu Gly Glu Pro Ala Gly Ile
Tyr 405 410 415Gln Tyr Ala
Arg Asn Leu Thr Tyr Val Leu Ile Tyr Asn Ala Ser His 420
425 430Met Val Pro Tyr Asp Leu Pro Arg Gln Ser
Arg Asp Met Leu Asp Arg 435 440
445Phe Met Asn Val Asp Ile Ala Ser Ile Gly Gly Ser Pro Ala Asp Ser 450
455 460Arg Ile Asp Gly Glu Lys Leu Pro
Gln Thr Ser Val Gly Gly His Pro465 470
475 480Asn Ser Thr Ala Ala Glu Glu Gln Glu Lys Glu Arg
Ile Lys Glu Thr 485 490
495Glu Trp Lys Ala Tyr Ala Lys Ser Gly Glu Ala Val Leu Leu Val Val
500 505 510Ile Ile Gly Val Leu Val
Trp Gly Phe Phe Ile Trp Arg Ser Arg Arg 515 520
525Arg His Gln Gly Tyr Arg Gly Val Trp His Lys Asp Met Ser
Gly Ser 530 535 540Ser Val Leu Glu Arg
Phe His Asn Lys Arg Thr Gly Gly Ala Asp Val545 550
555 560Glu Ala Gly Asp Phe Asp Glu Ala Glu Leu
Asp Asp Leu His Ser Pro 565 570
575Asp Leu Glu Arg Glu His Tyr Ala Val Gly Glu Asp Ser Asp Glu Asp
580 585 590Asp Ile Ser Arg Gln
His Ser Gln Gln Ala Ser Arg Ala Gly Gly Ser 595
600 605His Asn Leu Ser 610137531PRTAspergillus niger
137Met Phe Leu Ile Ser Pro Ala Val Thr Val Ala Ala Ala Leu Leu Leu1
5 10 15Ile Asn Gly Ala Gly Ala
Thr Gln Ser Glu Arg Ser Arg Ala Ala Ala 20 25
30His Phe Ser Lys Arg His Pro Thr Tyr Arg Ala Ala Thr
Arg Ala Gln 35 40 45Ser Ser Asn
Thr Ser Asp Tyr Arg Phe Phe Asn Asn Arg Thr Lys Pro 50
55 60His Leu Val Glu Ser Leu Pro Asp Val His Phe Asp
Val Gly Glu Met65 70 75
80Tyr Ser Gly Ser Ile Pro Ile Asp Asp Ser Asn Asn Gly Ser Arg Ser
85 90 95Leu Phe Tyr Ile Phe Gln
Pro Lys Ile Gly Glu Pro Ser Asp Asp Leu 100
105 110Thr Ile Tyr Leu Asn Gly Gly Pro Gly Cys Ser Ser
Glu Gln Gly Phe 115 120 125Phe Gln
Glu Asn Gly Arg Phe Thr Trp Gln Pro Gly Thr Tyr Ala Pro 130
135 140Val Ile Asn Glu Tyr Ser Trp Val Asn Leu Thr
Asn Met Leu Trp Val145 150 155
160Asp Gln Pro Val Gly Thr Gly Phe Ser Val Gly Asn Val Thr Ala Thr
165 170 175Asn Glu Glu Glu
Ile Ala Ala Asp Phe Leu Asp Phe Phe Glu Lys Phe 180
185 190Glu Asp Leu Tyr Gly Ile Lys Asn Phe Arg Ile
Phe Met Thr Gly Glu 195 200 205Ser
Tyr Ala Gly Arg Tyr Val Pro Tyr Ile Ser Ser Ala Met Leu Asp 210
215 220Lys Asn Asp Thr Thr Arg Phe Asn Leu Ser
Gly Ala Leu Leu Tyr Asp225 230 235
240Ala Cys Ile Gly Gln Trp Asp Tyr Ile Gln Ala Glu Leu Pro Ala
Tyr 245 250 255Pro Phe Val
Lys Gln His Ala Ser Leu Phe Asn Phe Asn Gln Ser Tyr 260
265 270Met Asn Glu Leu Glu Thr Thr Tyr Glu Glu
Cys Gly Tyr Lys Ala Tyr 275 280
285Phe Asp Glu Tyr Phe Ala Phe Pro Pro Ser Gly Ile Gln Pro Pro Lys 290
295 300Tyr Met Asn Tyr Ser Glu Cys Asp
Ile Tyr Asn Met Ile Tyr Tyr Glu305 310
315 320Ala Tyr Asn Pro Asn Pro Cys Phe Asn Pro Tyr Arg
Val Ile Asp Glu 325 330
335Cys Pro Leu Leu Trp Asp Val Leu Gly Trp Pro Thr Asp Leu Ala Tyr
340 345 350Glu Pro Ala Pro Thr Thr
Tyr Phe Asn Arg Ile Asp Val Lys Lys Ala 355 360
365Leu His Ala Pro Met Asp Val Glu Trp Glu Leu Cys Ser Tyr
Asp Leu 370 375 380Val Phe Ala Gly Gly
Asp Ala Asp Pro Gly Pro Glu Gln Gln Gly Asp385 390
395 400Asp Ser Pro Asn Pro Thr Glu Gly Val Leu
Pro Arg Val Ile Glu Ala 405 410
415Thr Asn Arg Val Leu Ile Ala Asn Gly Asp Trp Asp Tyr Leu Ile Ile
420 425 430Thr Asn Gly Thr Leu
Leu Ala Ile Gln Asn Met Thr Trp Asn Gly Gln 435
440 445Leu Gly Phe Gln Ser Ala Pro Ala Thr Pro Ile Asp
Ile Gln Met Pro 450 455 460Asp Leu Gln
Trp Val Glu Ile Phe Glu Ala Gln Glu Gly Tyr Gly Gly465
470 475 480Leu Asp Gly Pro Gln Gly Val
Met Gly Val Gln His Tyr Glu Arg Gly 485
490 495Leu Met Trp Ala Glu Thr Tyr Gln Ser Gly His Lys
Gln Ala Gln Asp 500 505 510Gln
Gly Arg Val Ser Tyr Arg His Leu Gln Trp Leu Leu Gly Gln Val 515
520 525Glu Ile Leu
530138531PRTAspergillus niger 138Met Leu Phe Arg Ser Leu Leu Ser Thr Ala
Val Leu Ala Val Ser Leu1 5 10
15Cys Thr Asp Asn Ala Ser Ala Ala Lys His Gly Arg Phe Gly Gln Lys
20 25 30Ala Arg Asp Ala Met Asn
Ile Ala Lys Arg Ser Ala Asn Ala Val Lys 35 40
45His Ser Leu Lys Ile Pro Val Glu Asp Tyr Gln Phe Leu Asn
Asn Lys 50 55 60Thr Lys Pro Tyr Arg
Val Glu Ser Leu Pro Asp Val His Phe Asp Leu65 70
75 80Gly Glu Met Tyr Ser Gly Leu Val Pro Ile
Glu Lys Gly Asn Val Ser 85 90
95Arg Ser Leu Phe Phe Val Phe Gln Pro Thr Ile Gly Glu Pro Val Asp
100 105 110Glu Ile Thr Ile Trp
Leu Asn Gly Gly Pro Gly Cys Ser Ser Leu Glu 115
120 125Ala Phe Leu Gln Glu Asn Gly Arg Phe Val Trp Gln
Pro Gly Thr Tyr 130 135 140Gln Pro Val
Glu Asn Pro Tyr Ser Trp Val Asn Leu Thr Asn Val Leu145
150 155 160Trp Val Asp Gln Pro Val Gly
Thr Gly Phe Ser Leu Gly Val Pro Thr 165
170 175Ala Thr Ser Glu Glu Glu Ile Ala Glu Asp Phe Val
Lys Phe Phe Lys 180 185 190Asn
Trp Gln Gln Ile Phe Gly Ile Lys Asn Phe Lys Ile Tyr Val Thr 195
200 205Gly Glu Ser Tyr Ala Gly Arg Tyr Val
Pro Tyr Ile Ser Ala Ala Phe 210 215
220Leu Asp Gln Asn Asp Thr Glu His Phe Asn Leu Lys Gly Ala Leu Ala225
230 235 240Tyr Asp Pro Cys
Ile Gly Gln Phe Asp Tyr Val Gln Glu Glu Ala Pro 245
250 255Val Val Pro Phe Val Gln Lys Asn Asn Ala
Leu Phe Asn Phe Asn Ala 260 265
270Ser Phe Leu Ala Glu Leu Glu Ser Ile His Glu Gln Cys Gly Tyr Lys
275 280 285Asp Phe Ile Asp Gln Tyr Leu
Val Phe Pro Ala Ser Gly Val Gln Pro 290 295
300Pro Lys Ala Met Asn Trp Ser Asp Pro Thr Cys Asp Val Tyr Asp
Ile305 310 315 320Val Asn
Asn Ala Val Leu Asp Pro Asn Pro Cys Phe Asn Pro Tyr Glu
325 330 335Ile Asn Glu Met Cys Pro Ile
Leu Trp Asp Val Leu Gly Phe Pro Thr 340 345
350Glu Val Asp Tyr Leu Pro Ala Gly Ala Ser Ile Tyr Phe Asp
Arg Ala 355 360 365Asp Val Lys Arg
Ala Met His Ala Pro Asn Ile Thr Trp Ser Glu Cys 370
375 380Ser Val Glu Ser Val Phe Val Gly Gly Asp Gly Gly
Pro Glu Gln Glu385 390 395
400Gly Asp Tyr Ser Ala Asn Pro Ile Glu His Val Leu Pro Gln Val Ile
405 410 415Glu Gly Thr Asn Arg
Val Leu Ile Gly Asn Gly Asp Tyr Asp Met Val 420
425 430Ile Leu Thr Asn Gly Thr Leu Leu Ser Ile Gln Asn
Met Thr Trp Asn 435 440 445Gly Lys
Leu Gly Phe Asp Thr Ala Pro Ser Thr Pro Ile Asn Ile Asp 450
455 460Ile Pro Asp Leu Met Tyr Asn Glu Val Phe Ile
Glu Asn Gly Tyr Asp465 470 475
480Pro Gln Gly Gly Gln Gly Val Met Gly Ile Gln His Tyr Glu Arg Gly
485 490 495Leu Met Trp Ala
Glu Thr Phe Gln Ser Gly His Met Gln Pro Gln Phe 500
505 510Gln Pro Arg Val Ser Tyr Arg His Leu Glu Trp
Leu Leu Gly Arg Arg 515 520 525Asp
Thr Leu 530139492PRTAspergillus niger 139Met Lys Gly Ala Ala Leu Ile
Pro Leu Ala Ala Gly Ile Pro Phe Ala1 5 10
15His Gly Leu Ser Leu His Lys Arg Asp Gly Pro Ala Val
Val Arg Met 20 25 30Pro Ile
Glu Arg Arg Ser Ala Gln Ser Leu Gln Lys Arg Asp Ser Thr 35
40 45Val Gly Val Thr Leu Gln Asn Trp Asp Ala
Thr Tyr Tyr Ala Val Asn 50 55 60Leu
Thr Leu Gly Thr Pro Ala Gln Lys Val Ser Leu Ala Leu Asp Thr65
70 75 80Gly Ser Ser Asp Leu Trp
Val Asn Thr Gly Asn Ser Thr Tyr Cys Ser 85
90 95Ile Asp Asn Leu Cys Thr Pro Tyr Gly Leu Tyr Asn
Ala Ser Glu Ser 100 105 110Ser
Thr Val Lys Thr Val Gly Thr His Leu Asn Asp Thr Tyr Ala Asp 115
120 125Gly Thr Asn Leu Tyr Gly Pro Tyr Val
Thr Asp Lys Leu Thr Ile Gly 130 135
140Asn Thr Thr Ile Asp Asn Met Gln Phe Gly Ile Ala Glu Ser Thr Thr145
150 155 160Ser Lys Arg Gly
Ile Ala Gly Val Gly Tyr Lys Ile Ser Thr Tyr Gln 165
170 175Ala Glu His Asp Asp Lys Val Tyr Ala Asn
Leu Pro Gln Ala Leu Val 180 185
190Asp Ser Gly Ala Ile Lys Ser Ala Ala Tyr Ser Ile Trp Leu Asp Ser
195 200 205Leu Glu Ala Ser Thr Gly Ser
Leu Leu Phe Gly Gly Val Asn Thr Ala 210 215
220Lys Tyr Lys Gly Asp Leu Gln Thr Leu Pro Ile Ile Pro Val Tyr
Gly225 230 235 240Lys Tyr
Tyr Ser Leu Ala Ile Ala Leu Thr Glu Leu Ser Val Ala Thr
245 250 255Asp Ser Asn Ser Ser Ser Phe
Thr Asp Ser Leu Pro Leu Ser Val Ser 260 265
270Leu Asp Thr Gly Thr Thr Met Thr Ala Leu Pro Ser Asp Leu
Val Asn 275 280 285Lys Val Tyr Asp
Ala Leu Asn Ala Thr Tyr Asp Lys Thr Tyr Asp Met 290
295 300Ala Tyr Ile Asp Cys Asp Thr Arg Glu Ala Asp Tyr
Asn Val Thr Tyr305 310 315
320Ser Phe Ser Gly Ala Thr Ile Thr Val Ser Met Ser Glu Leu Ile Ile
325 330 335Pro Ala Thr Glu Pro
Gly Trp Pro Asp Asn Thr Cys Val Leu Gly Leu 340
345 350Val Pro Ser Gln Pro Gly Val Asn Leu Leu Gly Asp
Thr Phe Leu Arg 355 360 365Ser Ala
Tyr Val Val Tyr Asp Leu Glu Asn Asn Glu Ile Ser Leu Ala 370
375 380Asn Thr Asn Phe Asn Pro Gly Asp Asp Asp Ile
Leu Glu Ile Gly Thr385 390 395
400Gly Thr Ser Ala Val Pro Gly Ala Thr Pro Val Pro Ser Ala Val Ser
405 410 415Ser Ala Thr Gly
Asn Gly Leu Ile Ser Ser Gly Thr Ala Val Pro Thr 420
425 430Leu Ser Gly Val Thr Ile Thr Ala Thr Ala Thr
Ala Thr Gly Ser Thr 435 440 445Gly
Thr Gly Ser Ser Gly Gly Ser Ser Ala Glu Ala Thr Ser Thr Ser 450
455 460Ser Glu Gly Ala Ala Ala Gln Ala Thr Ser
Asn Pro Met Asn Leu Leu465 470 475
480Pro Gly Leu Ala Gly Ile Gly Leu Leu Leu Ala Leu
485 490140611PRTAspergillus niger 140Met Leu Ser Ser Leu
Leu Ser Gln Gly Ala Ala Val Ser Leu Ala Val1 5
10 15Leu Ser Leu Leu Pro Ser Pro Val Ala Ala Glu
Ile Phe Glu Lys Leu 20 25
30Ser Gly Val Pro Asn Gly Trp Arg Tyr Ala Asn Asn Pro Gln Gly Asn
35 40 45Glu Val Ile Arg Leu Gln Ile Ala
Leu Gln Gln His Asp Val Ala Gly 50 55
60Phe Glu Gln Ala Val Met Asp Met Ser Thr Pro Gly His Ala Asp Tyr65
70 75 80Gly Lys His Phe Arg
Thr His Asp Glu Met Lys Arg Met Leu Leu Pro 85
90 95Ser Glu Thr Ala Val Asp Ser Val Arg Asp Trp
Leu Glu Ser Ala Gly 100 105
110Val His Asn Ile Gln Val Asp Ala Asp Trp Val Lys Phe His Thr Thr
115 120 125Val Asn Lys Ala Asn Ala Leu
Leu Asp Ala Asp Phe Lys Trp Tyr Val 130 135
140Ser Asp Ala Lys His Ile Arg Arg Leu Arg Thr Leu Gln Tyr Ser
Ile145 150 155 160Pro Asp
Ala Leu Val Ser His Ile Asn Met Ile Gln Pro Thr Thr Arg
165 170 175Phe Gly Gln Ile Gln Pro Asn
Arg Ala Thr Met Arg Ser Lys Pro Lys 180 185
190His Ala Asp Glu Thr Phe Leu Thr Ala Ala Thr Leu Ala Gln
Asn Thr 195 200 205Ser His Cys Asp
Ser Ile Ile Thr Pro His Cys Leu Lys Gln Leu Tyr 210
215 220Asn Ile Gly Asp Tyr Gln Ala Asp Pro Lys Ser Gly
Ser Lys Ile Gly225 230 235
240Phe Ala Ser Tyr Leu Glu Glu Tyr Ala Arg Tyr Ala Asp Leu Glu Arg
245 250 255Phe Glu Gln His Leu
Ala Pro Asn Ala Ile Gly Gln Asn Phe Ser Val 260
265 270Val Gln Phe Asn Gly Gly Leu Asn Asp Gln Leu Ser
Ser Ser Asp Ser 275 280 285Gly Glu
Ala Asn Leu Asp Leu Gln Tyr Ile Leu Gly Val Ser Ala Pro 290
295 300Val Pro Ile Thr Glu Tyr Ser Thr Gly Gly Arg
Gly Glu Leu Val Pro305 310 315
320Asp Leu Ser Ser Pro Asp Pro Asn Asp Asn Ser Asn Glu Pro Tyr Leu
325 330 335Asp Phe Leu Gln
Gly Ile Leu Lys Leu Asn Asn Ser Asp Leu Pro Gln 340
345 350Val Ile Ser Thr Ser Tyr Gly Glu Asp Glu Gln
Thr Ile Pro Val Pro 355 360 365Tyr
Ala Arg Thr Val Cys Asn Leu Tyr Ala Gln Leu Gly Ser Arg Gly 370
375 380Val Ser Val Ile Phe Ser Ser Gly Asp Ser
Gly Val Gly Ala Ala Cys385 390 395
400Leu Thr Asn Asp Gly Thr Asn Arg Thr His Phe Pro Pro Gln Phe
Pro 405 410 415Ala Ser Cys
Pro Trp Val Thr Ser Val Gly Ala Thr Ser Lys Thr Ser 420
425 430Pro Glu Gln Ala Val Ser Phe Ser Ser Gly
Gly Phe Ser Asp Leu Trp 435 440
445Pro Arg Pro Ser Tyr Gln His Ala Ala Val Gln Thr Tyr Leu Thr Lys 450
455 460His Leu Gly Asn Lys Phe Ser Gly
Leu Phe Asn Ala Ser Gly Arg Ala465 470
475 480Phe Pro Asp Val Ser Ala Gln Gly Val Asn Tyr Ala
Val Tyr Asp Lys 485 490
495Gly Met Leu Gly Gln Phe Asp Gly Thr Ser Cys Ser Ala Pro Thr Phe
500 505 510Ser Gly Val Ile Ala Leu
Leu Asn Asp Ala Arg Leu Arg Ala Gly Leu 515 520
525Pro Val Met Gly Phe Leu Asn Pro Phe Leu Tyr Gly Val Gly
Ser Glu 530 535 540Lys Gly Ala Leu Asn
Asp Ile Val Asn Gly Gly Ser Val Gly Cys Asp545 550
555 560Gly Arg Asn Arg Phe Gly Gly Thr Pro Asn
Gly Ser Pro Val Val Pro 565 570
575Phe Ala Ser Trp Asn Ala Thr Thr Gly Trp Asp Pro Val Ser Gly Leu
580 585 590Gly Thr Pro Asp Phe
Ala Lys Leu Lys Gly Val Ala Leu Gly Glu Glu 595
600 605Gly Gly Asn 610141478PRTAspergillus niger
141Met Trp Leu Phe Leu Val Cys Ser Ile Leu Leu Pro Leu Gly Val Val1
5 10 15Asn Ala Gln Ser Gln Tyr
Phe Asn Asn Lys Thr Lys Glu Phe Val Val 20 25
30Asn Gly Ser Ala Ile Pro Phe Val Asp Phe Asp Ile Gly
Glu Ser Tyr 35 40 45Ala Gly Tyr
Leu Pro Asn Thr Pro Ser Gly Ile Ser Ser Leu Tyr Phe 50
55 60Trp Phe Phe Pro Ser Ser Asp Pro Asp Ala Ser Asp
Glu Ile Thr Val65 70 75
80Trp Leu Asn Gly Gly Pro Gly Cys Ser Ser Leu Ala Gly Ile Met Leu
85 90 95Glu Asn Gly Pro Phe Leu
Trp Gln Pro Gly Thr Tyr Arg Pro Val Arg 100
105 110Asn Pro Tyr Ala Trp Asn Asn Leu Thr Asn Met Val
Tyr Ile Asp Gln 115 120 125Pro Ala
Gly Thr Gly Phe Ser Leu Gly Pro Ser Thr Val Val Ser Glu 130
135 140Phe Asp Val Ala Arg Gln Phe Met Asp Phe Trp
Arg Arg Phe Met Lys145 150 155
160Thr Phe Asp Leu Gln Asn Arg Lys Ile Tyr Leu Thr Gly Glu Ser Tyr
165 170 175Ala Gly Gln Tyr
Ile Pro Tyr Ile Ala Ser Gln Met Leu Asp Gln Asp 180
185 190Asp Asp Glu Tyr Phe Arg Val Ala Gly Ile Gln
Ile Asn Asp Pro Tyr 195 200 205Ile
Asn Glu Leu Pro Val Leu Gln Asp Val Ala Thr Val Asn Gln His 210
215 220Arg Ser Leu Phe Pro Phe Asn Asp Thr Phe
Met Ser Gln Ile Thr Lys225 230 235
240Leu Ser Asp Asp Cys Gly Tyr Thr Ser Phe Leu Asp Asp Ala Leu
Thr 245 250 255Phe Pro Pro
Arg Ser Gln Phe Pro Ser Val Pro Tyr Asn Ala Ser Cys 260
265 270Asn Ile Trp Asp Ile Ile Asn Asn Ala Ser
Leu Ala Leu Asn Pro Cys 275 280
285Phe Asn Arg Tyr His Ile Pro Asp Ala Cys Pro Thr Pro Trp Asn Pro 290
295 300Val Gly Gly Pro Ile Val Gly Leu
Gly Pro Thr Asn Tyr Phe Asn Arg305 310
315 320Ser Asp Val Gln Lys Ala Ile Asn Ala Tyr Pro Thr
Asp Tyr Phe Val 325 330
335Cys Lys Asp Gly Ile Phe Pro Thr Ala Asn Gly Leu Asp Thr Ser Pro
340 345 350Pro Ser Ser Leu Gly Pro
Leu Pro Arg Val Ile Glu Gln Thr Asn Asn 355 360
365Thr Ile Ile Ala His Gly Leu Met Asp Phe Glu Leu Leu Ala
Gln Gly 370 375 380Thr Leu Ile Ser Ile
Gln Asn Met Thr Trp Asn Gly Lys Gln Gly Phe385 390
395 400Glu Arg Glu Pro Val Glu Pro Leu Phe Val
Pro Tyr Gly Gly Ser Ser 405 410
415Gly Gly Gly Val Leu Gly Thr Ala His Thr Glu Arg Gly Leu Thr Phe
420 425 430Ser Thr Val Phe Ser
Ser Gly His Glu Ile Pro Glu Tyr Ala Pro Gly 435
440 445Ala Ala Tyr Arg Gln Leu Glu Phe Leu Leu Gly Arg
Val Ala Asn Leu 450 455 460Ser Thr Ile
Ile Glu Gln Val Gln Ile Thr Glu Gln Asn Gly465 470
475142210PRTAspergillus niger 142Met Ser Lys Leu Ser Ala Ala Ile
Ser Lys Leu Ser Leu Ser Thr Ile1 5 10
15Ala Thr Thr Leu Leu Leu Leu Thr Pro Pro Thr Thr Ala Tyr
Phe Tyr 20 25 30Lys Tyr Pro
Ala Leu Phe Val Tyr Lys Asp Thr Asn Cys Thr Asp Ile 35
40 45Ser Phe Ser Leu Val Tyr Pro Ser Leu Gly Asn
Cys Asn Gly Gly Tyr 50 55 60Tyr Asp
Tyr Ala Gly Ser Phe Gln Met Phe Asn Ile Asp Ala Ala Tyr65
70 75 80Thr Cys Asn Gly Ser Asp Ser
Thr Leu Met Phe Glu Met Tyr Asn Ser 85 90
95Ser Gly Ser Asp Cys Gly Asp Glu Ser Asp Leu Leu Phe
Arg Gln Pro 100 105 110Val Thr
Glu Glu Cys Thr Val Ala Asp Val Glu Ser Pro Gly Pro Leu 115
120 125Glu Met Pro Val Trp Phe Glu Leu Gly Ser
Leu Leu Gly Asn Cys Gly 130 135 140Gly
Met Ala Gly Thr Met Leu Phe Gly Val Gly Ile Leu Glu Gly Gly145
150 155 160Leu Glu Thr Lys Leu Tyr
Trp Lys Cys Tyr Ser Ser Arg Leu Asn Thr 165
170 175Ser Val Thr Val His Arg Leu Ser Leu Ile Leu Ser
Met Gly Cys Thr 180 185 190Ser
Val Ser Asp Ser Tyr Asn Glu Leu Ala Ala Ala His Tyr Tyr Glu 195
200 205Asp Leu 210143608PRTAspergillus
niger 143Met Arg His Leu Leu Ser Leu Leu Val Leu Leu Ile Ala Ser Ala Ala1
5 10 15Leu Val Ser Ala
Val Pro Ala Gly Ser Ile Ile Thr Pro Gln Pro Pro 20
25 30Val Glu Pro Val His Leu Leu Ser Ser Gln Pro
Ser Asp Pro Arg Arg 35 40 45Pro
Trp Ile Arg Leu Arg Asp Trp Ile Ile Glu Ser Ile Trp Gly Ile 50
55 60Glu Lys Pro Ala Ser Arg Arg Phe Pro Leu
Asn Asp Ser Pro Arg Asn65 70 75
80Arg Ser Pro Pro Ser Arg Ile Leu Ala Arg Tyr Gly Ser Asp Val
Val 85 90 95Leu Arg Phe
Ser Leu Arg Asn His Asp Glu Ala Glu Ala Leu Ala Gln 100
105 110Ala Ala Asp Ile Leu Phe Leu Asp Val Trp
Ala Ser Thr Pro Ala Phe 115 120
125Val Asp Ile Arg Leu Ala Glu Glu Val Thr Ala Tyr Thr Pro Leu Ile 130
135 140Asp Asn Leu Ala Glu Arg Ile Tyr
Thr Thr Tyr Pro Ser Lys Lys Pro145 150
155 160Ile Gly Leu Glu Gly Gln Ser Gly Phe Ala Ser Ser
Ser Arg Pro Ala 165 170
175Pro Lys Phe Gly Asp Leu Phe Phe His Glu Tyr Gln Pro Leu Ser Val
180 185 190Ile Ile Pro Trp Met Arg
Leu Leu Ala Ser Met Phe Pro Ser His Val 195 200
205Arg Met Ile Ser Val Gly Val Ser Tyr Glu Gly Arg Glu Ile
Pro Ala 210 215 220Leu Arg Leu Ser Ala
Gly Ser Ser Thr Ala Ala Ser Gly Pro Arg Lys225 230
235 240Thr Ile Ile Val Thr Gly Gly Ser His Ala
Arg Glu Trp Ile Gly Thr 245 250
255Ser Thr Val Asn His Val Met Tyr Thr Leu Ile Thr Lys Tyr Gly Lys
260 265 270Ser Lys Ala Val Thr
Arg Leu Leu Gln Asp Phe Asp Trp Ile Met Ile 275
280 285Pro Thr Ile Asn Pro Asp Gly Tyr Val Tyr Thr Trp
Glu Thr Asp Arg 290 295 300Leu Trp Arg
Lys Asn Arg Gln Arg Thr Ser Leu Arg Phe Cys Pro Gly305
310 315 320Ile Asp Leu Asp Arg Ala Trp
Gly Phe Glu Trp Asp Gly Gly Arg Thr 325
330 335Arg Ala Asn Pro Cys Ser Glu Asn Tyr Ala Gly Asp
Glu Pro Phe Glu 340 345 350Gly
Met Glu Ala Gln Gln Leu Ala Gln Trp Ala Leu Asn Glu Thr Gln 355
360 365Asn Asn Asn Ala Asp Ile Val Ser Phe
Leu Asp Leu His Ser Tyr Ser 370 375
380Gln Thr Ile Leu Tyr Pro Phe Ser Tyr Ser Cys Ser Ser Ile Pro Pro385
390 395 400Thr Leu Glu Ser
Leu Glu Glu Leu Gly Leu Gly Leu Ala Lys Ala Ile 405
410 415Arg Tyr Ala Thr His Glu Ile Tyr Asp Val
Thr Ser Ala Cys Glu Gly 420 425
430Ile Val Thr Ala Ser Ala Ala Asp Asn Asn Pro Gly Arg Phe Phe Pro
435 440 445Ile Gly Gly Asn Ser Gly Gly
Ser Ala Leu Asp Trp Phe Tyr His Gln 450 455
460Val His Ala Thr Tyr Ser Tyr Gln Ile Lys Leu Arg Asp Arg Gly
Ser465 470 475 480Tyr Gly
Phe Leu Leu Pro Ser Glu His Ile Ile Pro Thr Gly Lys Glu
485 490 495Ile Tyr Asn Val Val Leu Lys
Leu Gly Ser Phe Leu Ile Gly Gly Asp 500 505
510Ser Phe Asp Val Asp Trp Glu Ser Glu Leu Phe Asp Leu Ser
Lys Asp 515 520 525Glu Ser Asp Leu
Asp Ser Arg Tyr Ser Lys Ser Asn Asp Arg Ser Pro 530
535 540Ala Tyr Leu His Asn Ala Asn Gly Pro Leu Pro Asn
Ile Asp Glu Asp545 550 555
560Glu Asp Lys Glu Trp Val Met Val Glu Glu Glu Asp Tyr Thr Asp Asp
565 570 575Asp Asp Asp Asp Asp
Asp Asp Asp Glu Glu Glu Glu Glu Glu Glu Glu 580
585 590Asp Thr Tyr Trp Ala Thr Glu His Thr Tyr Glu Phe
Arg Arg Arg Arg 595 600
605144416PRTAspergillus niger 144Met Ala Phe Leu Lys Arg Ile Leu Pro Leu
Leu Ala Leu Ile Leu Pro1 5 10
15Ala Val Phe Ser Ala Thr Glu Gln Val Pro His Pro Thr Ile Gln Thr
20 25 30Ile Pro Gly Lys Tyr Ile
Val Thr Phe Lys Ser Gly Ile Asp Asn Ala 35 40
45Lys Ile Glu Ser His Ala Ala Trp Val Thr Glu Leu His Arg
Arg Ser 50 55 60Leu Glu Gly Arg Ser
Thr Thr Glu Asp Asp Leu Pro Ala Gly Ile Glu65 70
75 80Arg Thr Tyr Arg Ile Ala Asn Phe Ala Gly
Tyr Ala Gly Ser Phe Asp 85 90
95Glu Lys Thr Ile Glu Glu Ile Arg Lys His Asp His Val Ala Tyr Val
100 105 110Glu Gln Asp Gln Val
Trp Tyr Leu Asp Thr Leu Val Thr Glu Arg Arg 115
120 125Ala Pro Trp Gly Leu Gly Ser Ile Ser His Arg Gly
Gly Ser Ser Thr 130 135 140Asp Tyr Ile
Tyr Asp Asp Ser Ala Gly Glu Gly Thr Tyr Ala Tyr Val145
150 155 160Val Asp Thr Gly Ile Leu Ala
Thr His Asn Glu Phe Gly Gly Arg Ala 165
170 175Ser Leu Ala Tyr Asn Ala Ala Gly Gly Glu His Val
Asp Asp Val Gly 180 185 190His
Gly Thr His Val Ala Gly Thr Ile Gly Gly Lys Thr Tyr Gly Val 195
200 205Ser Lys Asn Ala His Leu Leu Ser Val
Lys Val Phe Val Gly Glu Ser 210 215
220Ser Ser Thr Ser Val Ile Leu Asp Gly Phe Asn Trp Ala Ala Asn Asp225
230 235 240Ile Val Ser Lys
Asn Arg Thr Ser Lys Ala Ala Ile Asn Met Ser Leu 245
250 255Gly Gly Gly Tyr Ser Tyr Ala Phe Asn Asn
Ala Val Glu Asn Ala Phe 260 265
270Asp Glu Gly Val Leu Ser Cys Val Ala Ala Gly Asn Glu Asn Arg Asp
275 280 285Ala Ala Arg Thr Ser Pro Ala
Ser Ala Pro Asp Ala Ile Thr Val Ala 290 295
300Ala Ile Asn Arg Ser Asn Ala Arg Ala Ser Phe Ser Asn Tyr Gly
Ser305 310 315 320Val Val
Asp Ile Phe Ala Pro Gly Glu Gln Val Leu Ser Ala Trp Thr
325 330 335Gly Ser Asn Ser Ala Thr Asn
Thr Ile Ser Gly Thr Ser Met Ala Thr 340 345
350Pro His Val Thr Gly Leu Ile Leu Tyr Leu Met Gly Leu Arg
Asp Leu 355 360 365Ala Thr Pro Ala
Ala Ala Thr Thr Glu Leu Lys Arg Leu Ala Thr Arg 370
375 380Asn Ala Val Thr Asn Val Ala Gly Ser Pro Asn Leu
Leu Ala Tyr Asn385 390 395
400Gly Asn Ser Gly Val Ser Lys Gly Gly Ser Asp Asp Gly Asp Glu Asp
405 410 415145455PRTAspergillus
niger 145Met Ile Thr Leu Leu Ser Ala Leu Phe Gly Ser Val Val Tyr Ala Ala1
5 10 15Thr Gln Thr Val
Leu Gly Pro Glu Gly Ala Asp Pro Phe Thr Val Phe 20
25 30Arg Ser Pro His Ser Pro Ala Phe Ser Ile Arg
Ile Gln Glu Gln Asn 35 40 45Asp
Ser Ile Cys Asp Ala Arg Ser Pro Gln Phe Thr Gly Trp Leu Asp 50
55 60Ile Gly Pro Lys His Leu Phe Phe Trp Tyr
Phe Glu Ser Gln Asn Asp65 70 75
80Pro Phe His Asp Pro Leu Thr Leu Trp Met Thr Gly Gly Pro Gly
Asp 85 90 95Ser Ser Met
Ile Gly Leu Phe Glu Glu Val Gly Pro Cys Arg Ile Asn 100
105 110Glu Phe Gly Asn Gly Thr Asp His Asn Pro
Trp Ala Trp Thr Lys Asn 115 120
125Ser Ser Leu Leu Phe Val Asp Gln Pro Val Asp Val Gly Phe Ser Tyr 130
135 140Ile Asp Glu Gly Tyr Glu Leu Pro
His Asp Ser Arg Glu Ala Ala Val145 150
155 160Asp Met His Arg Phe Leu Arg Leu Phe Ile Ser Glu
Ile Phe Pro His 165 170
175Lys Gln Phe Leu Pro Val His Leu Ser Gly Glu Ser Tyr Ala Gly Arg
180 185 190Tyr Ile Pro Tyr Leu Ala
Thr Gln Ile Leu Glu Gln Asn Glu Leu Tyr 195 200
205Lys Asp Ser Pro Arg Ile Pro Leu Lys Ser Cys Leu Val Gly
Asn Gly 210 215 220Phe Met Ser Pro Lys
Asp Ala Thr Phe Gly Tyr Trp Glu Thr Leu Cys225 230
235 240Thr Thr Asn Ser Gly Val Pro Ser Pro Ile
Phe Asn Glu Thr Arg Cys 245 250
255Asp Ile Met Ala Ala Asn Met Pro His Cys Met Asp Leu Tyr Asp Ile
260 265 270Cys Ile Gln His Ser
Asp Pro Ala Ile Cys His Ala Ala Gln Ser Val 275
280 285Cys Tyr Asp Ser Val Val Gly Leu Met Ala Lys Leu
Leu Leu Arg Met 290 295 300Thr Thr Val
Thr Ala Pro Cys Glu Ile Asp Glu Met Cys Tyr Ile Glu305
310 315 320Ala Ala Leu Ile Glu Arg Tyr
Leu Asn Ser Pro Ser Val Trp Glu Ala 325
330 335Leu Ser Pro Pro Gln Gln Val Thr Glu Tyr Lys Phe
Val Ala Thr Ser 340 345 350Val
Ile Asp Ala Phe Ala Gln Ser Ala Asp Gly Met Val Ser Ser Ser 355
360 365Lys Gln Ile Ala Phe Leu Leu Ala Asn
Asn Val Asp Phe Leu Ala Tyr 370 375
380Gln Gly Asn Leu Asp Leu Ala Cys Asn Thr Ala Gly Asn Leu Arg Trp385
390 395 400Ala Asn Ser Leu
Ser Trp Lys Gly Gln Thr Glu Phe Thr Ala Lys Pro 405
410 415Leu Leu Pro Trp Glu Ile Gln Val Ser Val
Gly Glu Gly Thr Asp Glu 420 425
430Thr Ser Arg Phe Ala Phe Val Thr Val Asp Asn Ala Gly His Leu Leu
435 440 445Arg Asp Ser Lys Ile Ser Asn
450 455146791PRTAspergillus niger 146Met Arg Phe Leu Thr
Tyr Ser Leu Pro Phe Ile Ala Ser Ala Ile Ser1 5
10 15Leu Phe Gly Val Asn Val Gln Ala Arg Ser Gln
Ala Pro Ser Ala Ile 20 25
30Arg His Val Ser Thr Leu Asp Gln Pro Thr Ile Lys Thr Pro Ser Gln
35 40 45Arg Val Asp His Leu Asp His Phe
Asp Ile Thr Phe Asn Ile His Asp 50 55
60Lys His Gln Arg Ile Lys Leu Glu Leu Glu Pro Asn His Asp Ile Leu65
70 75 80Ala Glu Asp Ala Ser
Val Gln Tyr Leu Asp Ala Asp Gly Asn Val Arg 85
90 95Arg His Glu Pro Ile Ala Pro His Glu His Lys
Val Phe Lys Gly Arg 100 105
110Ser Leu Leu Gly Arg Gly Lys Gly Met Trp Asp Pro Val Gly Trp Ala
115 120 125Arg Ile Tyr Leu Lys Gln Asp
Gly Ser Glu Pro Leu Phe Glu Gly Val 130 135
140Phe Ser Ile Asp Gly Asp Asn His His Val Gln Leu Lys Ser Ala
Tyr145 150 155 160Met Glu
Lys Lys Arg Pro Val Asp Val Asp Leu Pro Asp Ser Ala Thr
165 170 175Asp Tyr Met Ile Phe Tyr Arg
Asp Ser Asp Met Val Arg Leu His Thr 180 185
190Glu Leu Lys Arg Ser Ser Leu Gly Ser Thr Ser Cys Gln Ala
Asp Gln 195 200 205Leu Gly Phe Asn
Thr Asn Pro Asn His Pro Val Leu Gln Pro Tyr Gly 210
215 220Gln Ala Glu Thr Asp Thr Trp Gly Ala Ile Ser Leu
Asn Ser Leu Phe225 230 235
240Gly Leu Asn Lys Arg Gln Ser Asp Ile Gly Ser Val Ser Gly Asn Ala
245 250 255Gly Gly Val Asn Leu
Ala Ser Thr Ile Gly Asp Thr Ser Gly Cys Pro 260
265 270Ser Thr Lys Gln Val Ala Leu Ile Gly Val Ala Thr
Asp Cys Ala Phe 275 280 285Thr Gly
Ser Phe Asn Asn Glu Thr Ala Ala Lys Glu Trp Val Ile Ser 290
295 300Thr Val Asn Ser Ala Ser Asn Val Tyr Glu Lys
Ser Phe Asn Ile Thr305 310 315
320Ile Gly Leu Arg Asn Leu Thr Ile Thr Asp Ser Ser Cys Pro Asp Asn
325 330 335Pro Pro Ala Ala
Thr Ala Trp Asn Met Pro Cys Ser Ser Gly Asn Leu 340
345 350Thr Ser Arg Leu Asp Leu Phe Ser Lys Trp Arg
Gly Glu Gln Ser Asp 355 360 365Asp
Asn Ala Tyr Trp Thr Leu Met Ser Asp Cys Ala Thr Gly Asn Glu 370
375 380Val Gly Leu Ser Trp Leu Gly Gln Leu Cys
Asn Ser Asp Ala Ser Ser385 390 395
400Asp Gly Ser Ser Thr Val Ser Gly Thr Asn Val Val Val Arg Ser
Ser 405 410 415Gly Ser Asp
Trp Gln Ile Phe Ala His Glu Ser Gly His Thr Phe Gly 420
425 430Ala Val His Asp Cys Asp Ser Gln Thr Cys
Ala Glu Asp Leu Glu Ala 435 440
445Ser Ser Gln Cys Cys Pro Leu Thr Ser Ser Thr Cys Asn Ala Asn Gly 450
455 460Lys Tyr Ile Met Asn Pro Thr Thr
Gly Thr Asp Ile Thr Ala Phe Ser465 470
475 480Gln Cys Thr Ile Gly Asn Ile Cys Ala Ala Leu Gly
Arg Asn Ser Val 485 490
495Lys Ser Ser Cys Leu Ser Ala Asn Arg Asp Val Thr Thr Tyr Thr Gly
500 505 510Ser Gln Cys Gly Asn Gly
Ile Val Glu Ser Gly Glu Asp Cys Asp Cys 515 520
525Gly Gly Glu Asp Gly Cys Gly Asp Asn Asn Cys Cys Asp Ala
Lys Thr 530 535 540Cys Lys Phe Lys Ser
Gly Ala Val Cys Asp Asp Ser Asn Asp Ser Cys545 550
555 560Cys Ser Ser Cys Gln Phe Ser Ser Ala Gly
Thr Val Cys Arg Ala Ser 565 570
575Arg Gly Asp Cys Asp Val Ala Glu Thr Cys Ser Gly Asn Ser Ser Thr
580 585 590Cys Pro Thr Asp Ser
Phe Lys Lys Asp Gly Thr Ser Cys Gly Ser Ser 595
600 605Gly Ser Gly Leu Ala Cys Ala Ser Gly Gln Cys Thr
Ser Arg Asp Tyr 610 615 620Gln Cys Arg
Ser Val Met Gly Ser Leu Leu His Ser Asn Asp Thr Tyr625
630 635 640Ala Cys Ser Ser Phe Ser Ser
Ser Cys Glu Leu Val Cys Thr Ser Pro 645
650 655Lys Ile Gly Thr Cys Tyr Ser Val Asn Gln Asn Phe
Leu Asp Gly Thr 660 665 670Pro
Cys Gly Ser Gly Gly Tyr Cys Ser Asn Gly Asp Cys Lys Gly Gln 675
680 685Asn Val Glu Ser Trp Ile Lys Asn His
Lys Gly Ile Val Ile Gly Val 690 695
700Ala Cys Ala Val Gly Ala Leu Ile Leu Leu Ala Leu Met Thr Cys Ile705
710 715 720Val Asn Arg Cys
Arg Arg Ala Arg Ala Pro Lys Pro Val Pro Arg Pro 725
730 735Val Pro Tyr Gly Pro Trp Pro Gly Ala Arg
Pro Pro Pro Pro Pro Pro 740 745
750Met Asn Gln Trp Pro Ala Arg Gly Tyr Gln Gly Leu Gly Asn Glu Pro
755 760 765Pro Pro Pro Tyr Pro Gly Val
Pro Gly Gln Pro Val Pro Gln His Met 770 775
780Pro Pro Gln Gly Arg Tyr Ala785
790147481PRTAspergillus niger 147Met Arg Phe Leu Ser Ser Ala Ala Leu Phe
Gly Leu Ala Tyr Ala Ser1 5 10
15Thr Gln Ala Val Leu Gln Pro Glu Glu Pro Ser Asp Phe Arg Thr Phe
20 25 30His Ser Pro Tyr Ser Pro
His His Ser Ile Arg Ile Arg Gln Gln Asn 35 40
45Glu Ser Ile Cys Ala Ala His Ser Ala Gln Tyr Thr Gly Trp
Leu Asp 50 55 60Ile Gly Arg Lys His
Leu Phe Phe Trp Tyr Phe Glu Ser Gln Asn Asp65 70
75 80Pro Ala Asn Asp Pro Leu Thr Leu Trp Met
Thr Gly Gly Pro Gly Gly 85 90
95Ser Ser Met Ile Gly Leu Phe Glu Glu Val Gly Pro Cys Leu Ile Asn
100 105 110Glu Tyr Gly Asn Gly
Thr Tyr Tyr Asn Pro Trp Gly Trp Ser Arg Asn 115
120 125Ser Ser Leu Leu Phe Val Asp Gln Pro Val Asp Val
Gly Phe Ser Tyr 130 135 140Val Asp Glu
Gly Glu Asp Leu Pro Gly Asp Ser His Gln Ala Ala Ile145
150 155 160Asp Met His Arg Phe Leu Gln
Leu Phe Val Ser Glu Val Phe Pro Gln 165
170 175Leu Gln Thr Leu Pro Val His Leu Ser Gly Glu Ser
Tyr Ala Gly His 180 185 190Tyr
Val Pro Tyr Leu Gly Ser Gln Ile Val Gln Gln Asn Lys Leu Tyr 195
200 205Pro Thr Glu Pro Gln Val Leu Leu His
Ser Cys Leu Val Gly Asn Gly 210 215
220Tyr Tyr Ser Pro Arg Asp Thr Thr Tyr Gly Tyr Trp Glu Thr Leu Cys225
230 235 240Thr Thr Asn Pro
Gly Val Pro Glu Pro Val Phe Asn Arg Thr Arg Cys 245
250 255Asp Ile Met Ala Ala Asn Met Pro Arg Cys
Met Glu Val Ser Asp Val 260 265
270Cys Val Arg Asn Pro Asp Pro Ala Ile Cys His Ala Ala Ser Glu Val
275 280 285Cys Tyr Glu Gly Val Ile Gly
Trp Tyr Asp Asp Glu Ser Gly Glu Gly 290 295
300Gly Arg Asn Arg Phe Asp Ile Thr Ala Pro Cys Ala Leu Asp Gly
Ile305 310 315 320Cys Tyr
Ile Glu Ala Ala Arg Ile Glu Gln Tyr Leu Asn Thr Pro Ala
325 330 335Val Trp Ala Ala Leu Ser Pro
Pro Lys Glu Ile Lys Glu Tyr Lys Val 340 345
350Thr Ser Asp Asn Val Ser Arg Ala Phe Asp Leu Thr Ser Asp
Thr Met 355 360 365Thr Pro Ala Ser
Glu Gln Val Ala Phe Leu Leu Ala Asn Gln Val His 370
375 380Phe Leu Ala Tyr Gln Gly Asn Leu Asp Leu Ala Cys
Asn Thr Ala Gly385 390 395
400Asn Leu Arg Trp Ala His Ser Leu Pro Trp Arg Gly Gln Val Glu Phe
405 410 415Ala Ser Lys Ala Leu
Arg Pro Trp Ser Trp Val Asp Val Val Ser Gly 420
425 430Lys Gly Gly Val Ala Gly Thr Thr Lys Glu Glu Ser
Arg Phe Ala Leu 435 440 445Val Thr
Val Asp Gly Ala Gly His Phe Leu Pro Gln Asp Arg Pro Asp 450
455 460Ile Ala Leu Asp Met Met Val Arg Trp Ile Ser
Gly Ala Ser Phe Thr465 470 475
480Glu148319PRTAspergillus niger 148Met Thr Leu Leu Leu Asn Phe His
Ala Leu Phe Thr Val Ile Leu Val1 5 10
15Ala Asn Leu Ser Thr Arg Cys Ser Ala Leu Leu Ser Gly Arg
Asp Phe 20 25 30Cys Ser Thr
Pro Ala Pro Gly Glu Ser Leu Arg Ala Glu His Arg Arg 35
40 45Leu Tyr Asp Val Gln Ala Gln Arg Asp Ser Thr
Ala Glu Glu Ser Arg 50 55 60Glu Val
Val Pro Trp Ile Glu Ile Glu Thr Trp Phe His Ile Val Ser65
70 75 80Ser Asn Glu Ala Ala Asn Thr
Val Ser Asp Asp Met Ile Thr Ser Gln 85 90
95Leu Ser Tyr Leu Gln Lys Ala Tyr Glu Ser Ala Thr Ile
Thr Tyr Arg 100 105 110Leu Glu
Gly Ile Thr Arg His Ile Asn Asp Ser Trp Ala Arg Asn Asp 115
120 125Asp Glu Leu Gly Met Lys Asn Ala Leu Arg
Arg Gly Asn Tyr Gly Thr 130 135 140Leu
Asn Val Tyr Phe Gln Thr Asp Leu Gln Ala Ser Ser Asp Glu Asn145
150 155 160Ser Arg Asp Tyr Pro Asn
Asp Gly Asn Arg Arg Thr Asp Val Ser Asp 165
170 175Gln Ser Ser Ser Thr Val Leu Gly Phe Cys Thr Leu
Pro Asp Pro Ser 180 185 190Val
Asn Ser Ser Ser Pro Arg Ser Ser Tyr Ile Lys Asp Gly Cys Asn 195
200 205Val Leu Ala Asp Ile Met Pro Gly Gly
Ser Leu Ala Gln Tyr Asn Lys 210 215
220Gly Gly Thr Ala Val His Glu Val Gly His Trp Asn Gly Leu Leu His225
230 235 240Thr Phe Glu Gly
Glu Ser Cys Ser Pro Asp Asn Glu Gly Asp Tyr Ile 245
250 255Asp Asp Thr Pro Glu Gln Ser Glu Pro Thr
Ser Gly Cys Pro Ala Glu 260 265
270Lys Asp Ser Cys Pro Asp Leu Pro Gly Leu Asp Ala Ile His Asn Phe
275 280 285Met Asp Tyr Ser Ser Asp Asp
Cys Tyr Glu Ser Phe Thr Pro Asp Gln 290 295
300Ala Glu Arg Met Arg Ser Met Trp Ser Ala Met Arg Glu Gly Lys305
310 315149639PRTAspergillus niger 149Met His
Val Ser Leu Phe Leu Leu Ser Val Thr Ala Ala Phe Ala Ser1 5
10 15Pro Thr Pro His Asn Tyr Val Val
His Glu Arg Arg Asp Ala Leu Pro 20 25
30Ser Val Trp Val Glu Glu Ser Arg Leu Asp Lys Gly Ala Leu Leu
Pro 35 40 45Met Arg Ile Gly Leu
Thr Gln Ser Asn Leu Asp Arg Gly His Asp Leu 50 55
60Leu Met Glu Val Ser His Pro Gln Ser Ser Arg Tyr Gly Lys
His Leu65 70 75 80Ser
Ser Glu Glu Val His Asp Leu Phe Ala Pro Ser Asn Glu Ala Val
85 90 95Glu Thr Val Arg Thr Trp Ile
Glu Ser Ala Gly Ile Ala Pro Ser Arg 100 105
110Ile Ser Gln Ser Tyr Asn Lys Gln Trp Leu Gln Phe Asp Ala
His Ala 115 120 125Ser Glu Val Glu
Gln Leu Leu Gln Thr Glu Tyr Tyr Ile Tyr Thr His 130
135 140Ala Asp Thr Gly Ser Ser His Val Thr Cys His Glu
Tyr His Val Pro145 150 155
160Glu Thr Ile Gln Ser His Ile Asp Tyr Ile Thr Pro Gly Val Lys Met
165 170 175Leu Glu Val Arg Gly
Thr Pro Ser Lys Lys Arg Asp Ala Glu Lys Arg 180
185 190Ser Leu Gly Ser Leu Pro Pro Ile Leu Ala Pro Leu
Pro Ile Asn Ile 195 200 205Thr Lys
Ile Phe Asp Asp Pro Leu Ala His Cys Asp Leu Ala Val Thr 210
215 220Pro Asp Cys Ile Arg Ala Met Tyr Asn Ile Thr
Lys Gly Thr Thr Ala225 230 235
240Thr Lys Gly Asn Glu Leu Gly Ile Phe Glu Asp Leu Gly Asp Ile Tyr
245 250 255Ser Gln Asp Asp
Leu Asn Leu Phe Phe Ala Asn Phe Ala Ser Asp Ile 260
265 270Pro Gln Gly Thr His Pro Thr Leu Asp Ser Ile
Asp Gly Ala Thr Ala 275 280 285Pro
Thr Asp Val Thr Asn Ala Gly Pro Glu Ser Asp Leu Asp Phe Gln 290
295 300Ile Ala Tyr Pro Ile Ile Trp Pro Gln Asn
Thr Ile Leu Tyr Gln Thr305 310 315
320Asp Asp Pro Asn Tyr Glu Asp Asn Tyr Asn Phe Lys Gly Leu Leu
Asn 325 330 335Asn Phe Leu
Tyr Ala Ile Asp Gly Ser Tyr Cys Asn Glu Thr Ser Ser 340
345 350Leu Asp Pro Gln Tyr Pro Asp Pro Ser Pro
Gly Gly Tyr Ser Ser Pro 355 360
365Lys Gln Cys Gly Val Tyr Thr Pro Thr Asn Val Ile Ser Ile Ser Tyr 370
375 380Gly Ser Pro Glu Ala Asp Leu Pro
Ile Ala Tyr Gln Arg Arg Gln Cys385 390
395 400His Glu Phe Met Lys Leu Gly Leu Gln Gly Ile Ser
Val Val Val Ala 405 410
415Ser Gly Asp Ser Gly Val Ala Ser Ser Thr Gly Thr Cys Phe Gly Asp
420 425 430Ala Asp Asn Val Phe Val
Pro Asp Phe Pro Ala Thr Cys Pro Tyr Leu 435 440
445Thr Ala Val Gly Gly Thr Tyr Leu Pro Leu Gly Ala Asp Ala
Ala Lys 450 455 460Asp Gln Glu Ile Ala
Val Thr Arg Phe Pro Ser Gly Gly Gly Phe Ser465 470
475 480Asn Ile Tyr Ala Arg Pro Ser Tyr Gln Asn
His Ser Val Glu Thr Tyr 485 490
495Phe Ser Thr Thr Ser Asp Asp Leu Thr Tyr Pro Tyr Tyr Ser Gly Val
500 505 510Asn Tyr Thr Asp Phe
Ser Asn Thr Asp Gly Val Tyr Asn Arg Ile Gly 515
520 525Arg Gly Tyr Pro Asp Val Ser Ala Ile Ala Asp Asn
Ile Ile Ile Tyr 530 535 540Asn Gln Gly
Glu Ala Thr Leu Val Gly Gly Thr Ser Ala Ala Ala Pro545
550 555 560Ala Phe Ala Ala Met Leu Thr
Arg Ile Asn Glu Glu Arg Leu Ala Lys 565
570 575Gly Lys Ser Thr Val Gly Phe Val Asn Pro Val Leu
Tyr Glu His Pro 580 585 590Glu
Ala Phe Arg Asp Val Thr Val Gly Ser Asn Pro Gly Cys Gly Thr 595
600 605Asp Gly Phe Pro Val Ala Gly Gly Trp
Asp Pro Val Thr Gly Leu Gly 610 615
620Thr Pro Arg Phe Glu Asp Leu Met Asp Ile Phe Val Gly Asp Asp625
630 635150371PRTAspergillus niger 150Met Ala Ser
Lys Thr Leu Leu Leu Ile Pro Ala Leu Ala Thr Ala Ala1 5
10 15Leu Gly Ser Val Leu Asp Leu Asp Ile
Lys Val Asp Leu Gly Thr Pro 20 25
30Gly Gly Pro Phe Asp Leu Met Tyr Asp Thr Gly Ser Ser Thr Leu Trp
35 40 45Val Leu Asp Ser Asn Cys Thr
Asp Asp Cys Pro Asn Val Ser Gly Tyr 50 55
60Ser Arg His Gly Tyr Asn Leu Thr Ser Thr Gly Val Asn Leu Gly Val65
70 75 80Asn Asp Ser Ile
Ala Tyr Ser Gly Gly Thr Val Ser Gly Phe Thr Ala 85
90 95Thr Asp Ile Leu Thr Val Pro Asp Thr Asn
Val Ser Tyr Arg Gln Ser 100 105
110Phe Ala Val Ile Thr Asp Ser Thr Trp Ala Ala Leu Ala Ala Asp Gly
115 120 125Phe Ile Gly Leu Ala Ser Ser
Thr Ile Ala Phe Lys Asn Thr Thr Thr 130 135
140Ala Val Glu Gln Met Met Gln Asp Gly Leu Leu Asp Glu Pro Arg
Phe145 150 155 160Ala Ile
Tyr Ala Gly Ser Gly Glu Ser Thr Val Thr Asn Pro Asn Pro
165 170 175Glu Asn Asn Gly Val Phe Thr
Phe Gly Gly Ser His Glu Glu Thr Tyr 180 185
190Ala Asp Gly Glu Leu Gln Trp Met Lys Met Leu Ser Pro Phe
Glu Ile 195 200 205Tyr Lys Thr Asn
Leu Leu Gly Ile Gln Gly His Asn Asn Ser Asp Gly 210
215 220Gln Ala Leu Ser Ser Asp Val Leu Asn Trp Tyr Gly
Gln Thr Asn Leu225 230 235
240Phe Asn Val Ala Gly Ala Ser Ser Ile Ser Ile Pro Asn Asp Gln Ile
245 250 255Glu Ala Met Tyr Ala
Leu Thr Pro Phe Ser Tyr Ala Asp Ile Ser Ser 260
265 270Gly Tyr Arg Pro Leu Cys Ser Asp Phe Asn Asp Thr
Trp Ser Ile Ser 275 280 285Phe Thr
Met Gly Phe Tyr Gly Glu Gly Val Thr Phe Asn Leu Thr Gly 290
295 300Asp Gln Leu Ala Val Pro Gly Tyr Gln Asp Asp
Asp His Cys Phe Pro305 310 315
320Pro Phe Asn Pro Trp Asp Ser Tyr Asn Thr Ile Ile Gly Gln His Trp
325 330 335Leu Ser Asn Phe
Tyr Ala Val Phe Asp Phe Gly Ser Phe Asp Pro Glu 340
345 350Thr Tyr Asp Ile Arg Val Gly Leu Ala Pro Leu
Lys Lys Glu Tyr Leu 355 360 365Pro
Ser Ala 370151414PRTAspergillus niger 151Met Phe Pro Cys Ser Arg Ile
Trp Ser Leu Leu Val Ala Ala Ala Thr1 5 10
15Ala Ser Ala Val Pro Thr Ser Leu Ala Thr Thr His Leu
Gln Ser Val 20 25 30Asp Leu
Leu Leu Thr Arg Ser Ser Tyr Gly Phe Leu Thr Asp Ile Ala 35
40 45Leu Gly Thr Pro Gly Gln Ser Leu Pro Tyr
Leu Val Asp Trp Thr Trp 50 55 60Thr
Gly His Tyr Val Val Thr Thr Leu Cys Tyr Asn Asp Pro Thr Ala65
70 75 80Thr Tyr Asp Cys Leu Asn
Val Asp Gln Lys Ile Phe Asn Gln Thr Leu 85
90 95Ser Ser Thr Phe Ile Asn Gln Thr Asp Gln Tyr Gly
Tyr Leu Tyr Trp 100 105 110Asp
Pro Asn His Phe Tyr Phe Thr Glu Pro Ala Ala Ala Asp Val Ala 115
120 125Thr Asp Met Leu Arg Ile Gly Pro Thr
Ala Val Asn Thr Thr Ile Gln 130 135
140Ala Ala Asn Phe Val Phe Asn Glu Thr Ile Ser Ala Phe Pro Phe Ser145
150 155 160Gly Val Tyr Gly
Leu Ser Pro Val Phe Gln Gly Asp Asn Arg Ser Val 165
170 175Gln Ala Ser Phe Tyr Gln Gly Trp Arg Ser
Gly Ala Trp His Ser Pro 180 185
190Ile Val Ser Phe Ile Tyr Cys His Asp Asn Ala Thr Lys Ala Val Cys
195 200 205Ser Gly Tyr Asp Gly Leu Gln
Thr Leu Gly Gly Tyr Asn Thr Ser His 210 215
220Val Gln Gly Asp Ile Thr Trp Tyr Asp Ile Ile Val Thr Glu Ala
Ile225 230 235 240Asn Thr
Leu Asp Phe Val Tyr Ala Pro Ala Val Ile Asn Tyr Trp Ala
245 250 255Leu Asn Leu Thr Arg Phe Ser
Ile Gly Asp Glu Glu Gln Glu Leu Asn 260 265
270Lys Thr Thr Thr Leu Asp Gly Lys Gln Ala Ala Val Ala Ala
Phe Asp 275 280 285His Ala Ser Tyr
Gly Arg Gly Ala Pro Val Ser Val Tyr Gly Tyr Gln 290
295 300Arg Leu Val Glu Leu Val Gly Ala Lys Ala Val Thr
Leu Ser Asp Pro305 310 315
320Pro Asn Asn Gly Glu Gln Gly Phe Tyr Gln Phe Asp Cys Arg Asn Ser
325 330 335Ser Leu Leu Pro Pro
Leu Arg Tyr Glu Phe Ala Gly Ser Glu Arg Ala 340
345 350Trp Glu Ile Val Pro Glu Asn Tyr Val Glu Val Leu
Ala Asn Gly Thr 355 360 365Asn Lys
Cys Thr Phe Asn Val Arg Thr Leu Gly Asp Gly Ala Met Val 370
375 380Met Gly Asn Phe Gly Glu Thr Phe Ala Ile Asp
Lys Tyr Val Met Phe385 390 395
400Asp Phe Glu Lys Leu Gln Val Gly Ile Ala Asp Phe Ala Trp
405 410152480PRTAspergillus niger 152Met His Leu Pro
Gln Arg Leu Val Thr Ala Ala Cys Leu Cys Ala Ser1 5
10 15Ala Thr Ala Phe Ile Pro Tyr Thr Ile Lys
Leu Asp Thr Ser Asp Asp 20 25
30Ile Ser Ala Arg Asp Ser Leu Ala Arg Arg Phe Leu Pro Val Pro Lys
35 40 45Pro Ser Asp Ala Leu Ala Asp Asp
Ser Thr Ser Ser Ala Ser Asp Glu 50 55
60Ser Leu Ser Leu Asn Ile Lys Arg Ile Pro Val Arg Arg Asp Asn Asp65
70 75 80Phe Lys Ile Val Val
Ala Glu Thr Pro Ser Trp Ser Asn Thr Ala Ala 85
90 95Leu Asp Gln Asp Gly Ser Asp Ile Ser Tyr Ile
Ser Val Val Asn Ile 100 105
110Gly Ser Asp Glu Lys Ser Met Tyr Met Leu Leu Asp Thr Gly Gly Ser
115 120 125Asp Thr Trp Val Phe Gly Ser
Asn Cys Thr Ser Thr Pro Cys Thr Met 130 135
140His Asn Thr Phe Gly Ser Asp Asp Ser Ser Thr Leu Glu Met Thr
Ser145 150 155 160Glu Glu
Trp Ser Val Gly Tyr Gly Thr Gly Ser Val Ser Gly Leu Leu
165 170 175Gly Lys Asp Lys Leu Thr Ile
Ala Asn Val Thr Val Arg Met Thr Phe 180 185
190Gly Leu Ala Ser Asn Ala Ser Asp Asn Phe Glu Ser Tyr Pro
Met Asp 195 200 205Gly Ile Leu Gly
Leu Gly Arg Thr Asn Asp Ser Ser Tyr Asp Asn Pro 210
215 220Thr Phe Met Asp Ala Val Ala Glu Ser Asn Val Phe
Lys Ser Asn Ile225 230 235
240Val Gly Phe Ala Leu Ser Arg Ser Pro Ala Lys Asp Gly Thr Val Ser
245 250 255Phe Gly Thr Thr Asp
Lys Asp Lys Tyr Thr Gly Asp Ile Thr Tyr Thr 260
265 270Asp Thr Val Gly Ser Asp Ser Tyr Trp Arg Ile Pro
Val Asp Asp Val 275 280 285Tyr Val
Gly Gly Thr Ser Cys Asp Phe Ser Asn Lys Ser Ala Ile Ile 290
295 300Asp Thr Gly Thr Ser Tyr Ala Met Leu Pro Ser
Ser Asp Ser Lys Thr305 310 315
320Leu His Ser Leu Ile Pro Gly Ala Lys Ser Ser Gly Ser Tyr His Ile
325 330 335Ile Pro Cys Asn
Thr Thr Thr Lys Leu Gln Val Ala Phe Ser Gly Val 340
345 350Asn Tyr Thr Ile Ser Pro Lys Asp Tyr Val Gly
Ala Thr Ser Gly Ser 355 360 365Gly
Cys Val Ser Asn Ile Ile Ser Tyr Asp Leu Phe Gly Asp Asp Ile 370
375 380Trp Leu Leu Gly Asp Thr Phe Leu Lys Asn
Val Tyr Ala Val Phe Asp385 390 395
400Tyr Asp Glu Leu Arg Val Gly Phe Ala Glu Arg Ser Ser Asn Thr
Thr 405 410 415Ser Ala Ser
Asn Ser Thr Ser Ser Gly Thr Ser Ser Thr Ser Gly Ser 420
425 430Thr Thr Thr Gly Ser Ser Thr Thr Thr Thr
Ser Ser Ala Ser Ser Ser 435 440
445Ser Ser Ser Asp Ala Glu Ser Gly Ser Ser Met Thr Ile Pro Ala Pro 450
455 460Gln Tyr Phe Phe Ser Ala Leu Ala
Ile Ala Ser Phe Met Leu Trp Leu465 470
475 480153466PRTAspergillus niger 153Met Thr Ser Ser Thr
Leu Arg Leu Ala Val Ala Leu Ala Leu Ser Thr1 5
10 15Cys Ser Ser Ala Leu Ser Ser Gln Arg Asp Asp
Ser Leu Val Val Pro 20 25
30Phe Pro Phe Gly Asn Leu Glu Asp Val His Ile Ala Lys Arg Asp Ser
35 40 45Ser Lys Thr Val Glu Ala Pro Leu
Val Ile Tyr Gly Asp Ser Tyr Trp 50 55
60Met Asn Ala Ser Ile Gly Thr Pro Ala Gln Ser Leu Ser Phe Leu Leu65
70 75 80Asp Leu Thr Arg Ser
Arg Val Glu Pro Ala Tyr Thr Leu Asp Glu Asn 85
90 95Tyr Glu Cys Ser Asp Asp Glu Leu Cys Ser Glu
Phe Gly Phe Tyr Lys 100 105
110Pro Thr Asp Ser Ser Thr Tyr Gln His Leu Thr Tyr Thr Gln Arg His
115 120 125Asp Ala Gly Val Asp Tyr Ser
Tyr Leu Asp Thr Ile Thr Leu Gly Asp 130 135
140His Ala Thr Asp Asn Val Pro Leu Asp Met Tyr Leu Leu Ser Tyr
Ile145 150 155 160Ser Tyr
Ser Ser Leu Gly Leu Ser Ser Val Asn Thr Ser Phe Pro Tyr
165 170 175Ile Leu Val Asp Arg Gly Leu
Thr Thr Ser Pro Ser Phe Ser Leu Ile 180 185
190Gly Asp Asn Gly Asn Thr Thr Thr Pro Ser Ile Ile Phe Gly
Gly Ile 195 200 205Asn Thr Ser Lys
Phe Asn Gly Pro Leu Gln Ala Phe Ser Phe Ala Asp 210
215 220His Ser Ile Thr Asn Asn Pro Phe Val Thr Val Glu
Ala Asp Ser Leu225 230 235
240Gln Leu Thr Thr Asn Thr Asn Asp Asn Ser Thr Tyr Pro Ile Pro Ser
245 250 255Ser Thr Pro Met Met
Leu Arg Thr Glu Glu Leu Ile Thr Tyr Leu Pro 260
265 270Asn Ser Thr Val Gln Ser Leu Tyr Thr Asp Leu Asn
Ile Thr Met Asp 275 280 285Gly Val
Ile Ser Thr Ser Arg Phe Tyr Gly Val Leu Pro Cys Ala Arg 290
295 300Gln Glu Thr Glu Ser His Thr Ile Ser Leu Ala
Ile Gly Asn Met Thr305 310 315
320Phe Ser Val Ser Trp Asp Glu Leu Phe Val Pro Trp Thr Arg Asp Gly
325 330 335Leu Cys Lys Phe
Gly Ile Gln Ala Gln Asp Ser Asp Tyr Lys Thr Arg 340
345 350Ala Glu Leu Gly Val Pro Phe Leu Arg Arg Met
Tyr Val Ala Val Asp 355 360 365Tyr
Asn Asn Gln Phe Val Gly Val Ala Thr Leu Lys Asp Asp Asp Asp 370
375 380Gln Asn Gly Gly Glu Asp Glu Ile Val Glu
Ile Gly Thr Gly Thr Ala385 390 395
400Leu Pro Ser Ala Val Gly Asp Trp Pro Ala Ser Val Thr Ala Tyr
Thr 405 410 415Pro Ala Ala
Ser Thr Gly Thr Ala Ala Ala Thr Leu Thr Phe Thr Thr 420
425 430Ala Thr Ser Ser Gly Gly Gly Val Val Pro
Thr Gly Leu Ser Glu Leu 435 440
445Gly Arg Ala Phe Leu Val Pro Gly Val Leu Gly Met Ala Val Leu Gln 450
455 460Ala Val465154543PRTAspergillus
niger 154Met Met Arg Pro Ile Leu Leu Pro Leu Leu Gly Val Phe Leu Gln Thr1
5 10 15Ser Ser Ala Ser
Asn Pro Tyr Val Met Ser Trp Ser Ser Gln Ala Tyr 20
25 30Gly Pro Asp Gly Pro Trp Gln Ala Val Ser Ile
Asp Val Gly Ser Asn 35 40 45Gln
Gln Thr Val Asp Leu Tyr Pro Gly Ala Asn Tyr Ala Ser Thr Ile 50
55 60Leu Met Ser Thr Leu Cys Thr Asn Lys Thr
Leu Ser Ser Thr Cys Tyr65 70 75
80Ala Ala Glu Ala Gly Thr Phe Asn Gln Asn Thr Ser Thr Thr Ala
Tyr 85 90 95Thr Thr Ala
Ser Ser Trp Glu Thr Thr Tyr Trp Ala Val Glu Gly Gly 100
105 110Ser Gln Glu Ala Val Leu Gly Asp Glu Val
Thr Leu Gly Ser Phe Val 115 120
125Val Pro Asn Val Ser Phe Glu Ala Ile Tyr Gln Thr Tyr Gln Thr Tyr 130
135 140Pro Asn Gly Ile Ala Tyr Pro Val
Ser Val Gly Ser Leu Ala Leu Gly145 150
155 160Gly Pro Tyr Leu Ser Asp Thr Val Ser Asn Ser Thr
Val Leu Asn Met 165 170
175Ile Ala Gly Trp Leu Tyr Ser Ser Asn Asp Ile Pro Ser Tyr Ser Tyr
180 185 190Gly Met His Ile Gly Ser
Val Asp Pro Lys Ile Pro Gly Ser Leu Ile 195 200
205Leu Gly Gly Tyr Asp Lys Ser Arg Val Ile Gly Asp Val Ser
Ala Gln 210 215 220Gly Val Val Ser Ser
Ser Gly Leu Leu Glu Leu Glu Leu Lys Asp Ile225 230
235 240Gly Leu Gly Val Ala Ala Gly Ser Ser Pro
Phe Ser Phe Asn Asn Glu 245 250
255Ser Gly Leu Phe Leu Gln Ser Ser Gly Ser Val Gln Ala Lys Thr Val
260 265 270Gln Ile Asp Pro Thr
Lys Pro Tyr Met Tyr Leu Pro Gln Ala Thr Cys 275
280 285Asp Ala Ile Thr Ser Thr Met Pro Ile Ser Phe Asn
Ser Ser Leu Gly 290 295 300Leu Tyr Phe
Trp Asp Thr Thr Ser Asp Asp Tyr Leu Asn Ile Thr Ser305
310 315 320Ser Ala Ala Tyr Leu Ser Phe
Val Phe Asn Met Asn Gly Val Asn Asn 325
330 335Lys Asn Ile Thr Ile Lys Ile Pro Phe Ser Gln Leu
Asn Leu Thr Leu 340 345 350Gln
Glu Pro Leu Val Asp Gln Asn Val Thr Tyr Phe Pro Cys Phe Leu 355
360 365Thr Thr Ser Thr Pro Val Leu Gly Arg
Ala Phe Leu Gln Ser Ala Phe 370 375
380Val Gly Val Asn Trp Phe Asn Gly Asn Asn Ser Gly Thr Trp Phe Leu385
390 395 400Ala Gln Ala Pro
Gly Pro Gly Tyr Ala Ser Glu Asp Ile Thr Arg Ile 405
410 415Ala Val Ser Asp Thr Ser Leu Ser Ala Ser
Asn Gly Thr Trp Glu Glu 420 425
430Thr Trp Ala Thr Tyr Trp Gly Ile Lys Thr Ser Asp Asn Ser Ser Ser
435 440 445Ser Lys Ser Gly Leu Ser Ser
Gly Ala Lys Ile Gly Ile Gly Val Gly 450 455
460Val Gly Val Gly Gly Ala Val Leu Ile Ala Ala Gly Ile Ala Ile
Ala465 470 475 480Phe Cys
Leu Arg Arg Arg Arg Gly Ala Ser Gln Glu Ala Ala Gly Glu
485 490 495Gln Arg Arg Ser Met Phe Arg
Gly Phe Ala Glu Leu Pro Gly Gly Ala 500 505
510His Ser Glu Pro Ala Lys Glu Leu Asp Thr Lys Met His Lys
Pro Pro 515 520 525Gln Glu Met Met
Ala Ser Gln Glu Val Glu Arg Tyr Glu Leu Gly 530 535
540155844PRTAspergillus niger 155Met Arg Leu Thr Gly Gly Val
Ala Ala Ala Leu Gly Leu Cys Ala Ala1 5 10
15Ala Ser Ala Ser Leu His Pro His Arg Ser Tyr Glu Thr
His Asp Tyr 20 25 30Phe Ala
Leu His Leu Asp Glu Ser Thr Ser Pro Ala Asp Val Ala Gln 35
40 45Arg Leu Gly Ala Arg His Glu Gly Pro Val
Gly Glu Leu Pro Ser His 50 55 60His
Thr Phe Ser Ile Pro Arg Glu Asn Ser Asp Asp Val His Ala Leu65
70 75 80Leu Asp Gln Leu Arg Asp
Arg Arg Arg Leu Arg Arg Arg Ser Gly Asp 85
90 95Asp Ala Ala Val Leu Pro Ser Leu Val Gly Arg Asp
Glu Gly Leu Gly 100 105 110Gly
Ile Leu Trp Ser Glu Lys Leu Ala Pro Gln Arg Lys Leu His Lys 115
120 125Arg Val Pro Pro Thr Gly Tyr Ala Ala
Arg Ser Pro Val Asn Thr Gln 130 135
140Asn Asp Pro Gln Ala Leu Ala Ala Gln Lys Arg Ile Ala Ser Glu Leu145
150 155 160Gly Ile Ala Asp
Pro Ile Phe Gly Glu Gln Trp His Leu Tyr Asn Thr 165
170 175Val Gln Leu Gly His Asp Leu Asn Val Thr
Gly Ile Trp Leu Glu Gly 180 185
190Val Thr Gly Gln Gly Val Thr Thr Ala Ile Val Asp Asp Gly Leu Asp
195 200 205Met Tyr Ser Asn Asp Leu Arg
Pro Asn Tyr Phe Ala Ala Gly Ser Tyr 210 215
220Asp Tyr Asn Asp Lys Val Pro Glu Pro Arg Pro Arg Leu Ser Asp
Asp225 230 235 240Arg His
Gly Thr Arg Cys Ala Gly Glu Ile Gly Ala Ala Lys Asn Asp
245 250 255Val Cys Gly Val Gly Val Ala
Tyr Asp Ser Arg Ile Ala Gly Ile Arg 260 265
270Ile Leu Ser Ala Pro Ile Asp Asp Thr Asp Glu Ala Ala Ala
Ile Asn 275 280 285Tyr Ala Tyr Gln
Glu Asn Asp Ile Tyr Ser Cys Ser Trp Gly Pro Tyr 290
295 300Asp Asp Gly Ala Thr Met Glu Ala Pro Gly Thr Leu
Ile Lys Arg Ala305 310 315
320Met Val Asn Gly Ile Gln Asn Gly Arg Gly Gly Lys Gly Ser Val Phe
325 330 335Val Phe Ala Ala Gly
Asn Gly Ala Ile His Asp Asp Asn Cys Asn Phe 340
345 350Asp Gly Tyr Thr Asn Ser Ile Tyr Ser Ile Thr Val
Gly Ala Ile Asp 355 360 365Arg Glu
Gly Asn His Pro Pro Tyr Ser Glu Ser Cys Ser Ala Gln Leu 370
375 380Val Val Ala Tyr Ser Ser Gly Ala Ser Asp Ala
Ile His Thr Thr Asp385 390 395
400Val Gly Thr Asp Lys Cys Ser Thr Thr His Gly Gly Thr Ser Ala Ala
405 410 415Gly Pro Leu Ala
Ala Gly Thr Val Ala Leu Ala Leu Ser Val Arg Pro 420
425 430Glu Leu Thr Trp Arg Asp Val Gln Tyr Leu Met
Ile Glu Ala Ala Val 435 440 445Pro
Val His Glu Asp Asp Gly Ser Trp Gln Asp Thr Lys Asn Gly Lys 450
455 460Lys Phe Ser His Asp Trp Gly Tyr Gly Lys
Val Asp Thr Tyr Thr Leu465 470 475
480Val Lys Arg Ala Glu Thr Trp Asp Leu Val Lys Pro Gln Ala Trp
Leu 485 490 495His Ser Pro
Trp Gln Arg Val Glu His Glu Ile Pro Gln Gly Glu Gln 500
505 510Gly Leu Ala Ser Ser Tyr Glu Val Thr Glu
Asp Met Leu Lys Gly Ala 515 520
525Asn Leu Glu Arg Leu Glu His Val Thr Val Thr Met Asn Val Asn His 530
535 540Thr Arg Arg Gly Asp Leu Ser Val
Glu Leu Arg Ser Pro Asp Gly Arg545 550
555 560Val Ser His Leu Ser Thr Pro Arg Arg Pro Asp Asn
Gln Glu Val Gly 565 570
575Tyr Val Asp Trp Thr Phe Met Ser Val Ala His Trp Gly Glu Ser Gly
580 585 590Ile Gly Lys Trp Thr Val
Ile Val Lys Asp Thr Asn Val Asn Glu His 595 600
605Thr Gly Gln Phe Ile Asp Trp Arg Leu Asn Leu Trp Gly Glu
Ala Ile 610 615 620Asp Gly Ala Glu Gln
Pro Leu His Pro Met Pro Thr Glu His Asp Asp625 630
635 640Asp His Ser Tyr Glu Glu Gly Asn Val Ala
Thr Thr Ser Ile Ser Ala 645 650
655Val Pro Thr Lys Thr Glu Leu Pro Asp Lys Pro Thr Gly Gly Val Asp
660 665 670Arg Pro Val Asn Val
Lys Pro Thr Thr Ser Ala Met Pro Thr Gly Ser 675
680 685Leu Thr Glu Pro Ile Asp Asp Glu Glu Leu Gln Lys
Thr Pro Ser Thr 690 695 700Glu Ala Ser
Ser Thr Pro Ser Pro Ser Pro Thr Thr Ala Ser Asp Ser705
710 715 720Ile Leu Pro Ser Phe Phe Pro
Thr Phe Gly Ala Ser Lys Arg Thr Gln 725
730 735Val Trp Ile Tyr Ala Ala Ile Gly Ser Ile Ile Val
Phe Cys Ile Gly 740 745 750Leu
Gly Val Tyr Phe His Val Gln Arg Arg Lys Arg Ile Arg Asp Asp 755
760 765Ser Arg Asp Asp Tyr Asp Phe Glu Met
Ile Glu Asp Glu Asp Glu Leu 770 775
780Gln Ala Met Asn Gly Arg Ser Asn Arg Ser Arg Arg Arg Gly Gly Glu785
790 795 800Leu Tyr Asn Ala
Phe Ala Gly Glu Ser Asp Glu Glu Pro Leu Phe Ser 805
810 815Asp Glu Asp Asp Glu Pro Tyr Arg Asp Arg
Gly Ile Ser Gly Glu Gln 820 825
830Glu Arg Glu Gly Ala Asp Gly Glu His Ser Arg Arg 835
840156149PRTAspergillus niger 156Met Lys Thr Phe Ser Thr Val Thr Ser
Leu Leu Ala Leu Phe Ser Ser1 5 10
15Ala Leu Ala Ala Pro Val Asp Ser Ala Glu Ala Ala Gly Thr Thr
Val 20 25 30Ser Val Ser Tyr
Asp Thr Ala Tyr Asp Val Ser Gly Ala Ser Leu Thr 35
40 45Thr Val Ser Cys Ser Asp Gly Ala Asn Gly Leu Ile
Asn Lys Gly Tyr 50 55 60Ser Asn Phe
Gly Ser Leu Pro Gly Phe Pro Lys Ile Gly Gly Ala Pro65 70
75 80Thr Ile Ala Gly Trp Asn Ser Pro
Asn Cys Gly Lys Cys Tyr Ala Leu 85 90
95Thr Tyr Asn Gly Gln Thr Val Asn Ile Leu Ala Ile Asp Ser
Ala Pro 100 105 110Gly Gly Phe
Asn Ile Ala Leu Glu Ala Met Asn Thr Leu Thr Asn Asn 115
120 125Gln Ala Gln Gln Leu Gly Arg Ile Glu Ala Thr
Tyr Thr Glu Val Asp 130 135 140Val Ser
Leu Cys Ala145157296PRTAspergillus niger 157Met Ala Gln Ile Phe Trp Leu
Ser Leu Phe Leu Leu Val Ser Trp Val1 5 10
15Arg Ala Glu Ser Asn Arg Thr Glu Val Asp Leu Ile Phe
Pro Arg Asn 20 25 30Asp Thr
Phe Ala Pro Met Pro Leu Met Pro Val Val Phe Ala Val Gln 35
40 45Ala Pro Ser Val Ala His Lys Val Asn Thr
Tyr Ile Glu Tyr Gly Tyr 50 55 60Tyr
Pro Val Gly Arg Pro Asn Glu Thr Val Ile Gly Gln Thr Asp His65
70 75 80Val Ser Asp Ser Thr Asn
Glu Thr Thr Tyr Phe Ser Val Ser Gly Ile 85
90 95Gly Arg Thr Phe Asn Thr Thr Gly Ser Trp Glu Leu
Phe Trp Arg Leu 100 105 110Arg
Trp Thr Asn Cys Ser Ile Ser Glu Asp Ser Arg Tyr Tyr Asn Gln 115
120 125Ser Tyr Pro Trp Ile Ser Ser Pro Tyr
Ile Asp Gly Ser Leu Asn Ile 130 135
140Asp Lys Val Tyr Glu Gly Phe His Tyr Thr Ala Tyr Asn Val Ile Val145
150 155 160Asp Arg Val Thr
Phe Ser Thr Arg Glu Asp Ala Ser Gln Pro Asn Leu 165
170 175Thr Thr Leu Thr Asn Ser Glu Asn Cys Asp
Lys Val Ser Ser Leu Ala 180 185
190Leu Leu Ser Ile Val Asp Ser Leu Arg Ile Pro Pro Gln Leu Pro Gln
195 200 205Glu Asp Ile Asp Thr Val Ser
Met Cys Pro Gln Leu Ala Asp Ala Arg 210 215
220Leu Asn Ser Thr Ser Thr Ser Ser Pro Cys Ser Val Ser Ile Ser
Pro225 230 235 240Glu Val
Glu Ser Asn Ile Leu Ala Lys Ile Ala Asp Asn Glu Cys Asn
245 250 255Asn Ala Leu His Pro Ala Val
Ser Cys Thr Thr Glu Glu Thr Lys Glu 260 265
270Gly Ser Ala Ser Ser His Asp His Gly His Ala Val Trp Leu
Val Ile 275 280 285Thr Leu Ala Phe
Ala Phe Leu Phe 290 295158310PRTAspergillus niger
158Met Gly Gly Arg Asp Val Ala Ile Leu Ser Arg His Phe Ala Val Thr1
5 10 15Ser Ser Gln Ser Val Asn
Gly Val Val Ser Gly Met Phe Gln His Thr 20 25
30Val Thr Ser Ser Pro Ser Phe Thr Thr Asn Gln Phe Phe
Lys Lys Lys 35 40 45Phe Thr Ala
Ala Ile Ala Thr Ala Ile Phe Ala Ser Val Ala Val Ala 50
55 60Ala Pro Gln Arg Gly Leu Glu Ala Arg Leu Lys Ala
Arg Gly Ser Ser65 70 75
80Lys Gly Ser Arg Pro Leu Gln Ala Val Ala Arg Pro Ala Ser Thr Lys
85 90 95Asn Gln Thr Asn Val Glu
Tyr Ser Ser Asn Trp Ser Gly Ala Val Leu 100
105 110Val Glu Pro Pro Ser Ala Ala Ala Thr Tyr Thr Ala
Val Thr Gly Thr 115 120 125Phe Thr
Val Pro Glu Pro Thr Gly Asn Ser Gly Gly Ser Gln Ala Ala 130
135 140Ser Ala Trp Val Gly Ile Asp Gly Asp Thr Tyr
Gly Asn Ala Ile Leu145 150 155
160Gln Thr Gly Val Asp Phe Thr Val Thr Asp Gly Glu Ala Ser Phe Asp
165 170 175Ala Trp Tyr Glu
Trp Tyr Pro Asp Tyr Ala Tyr Asp Phe Ser Gly Ile 180
185 190Asp Ile Ser Ala Gly Asp Glu Ile Val Ala Ile
Val Glu Ser Tyr Thr 195 200 205Ser
Thr Thr Gly Ile Ala Ile Ile Glu Asn Lys Ser Thr Gly Gln Lys 210
215 220Val Ser Lys Glu Leu Ser Ser Ser Ser Ser
Leu Gly Gly Gln Asn Ala225 230 235
240Glu Trp Ile Val Glu Asp Phe Glu Glu Asn Gly Ser Leu Val Asn
Leu 245 250 255Val Asp Phe
Gly Thr Val Thr Phe Thr Gly Ala Val Ala Lys Ala Ala 260
265 270Gly Gly Glu Ser Val Gly Leu Thr Asp Ala
Thr Ile Ile Glu Ile Glu 275 280
285Glu Asn Gly Gln Val Val Thr Asp Val Thr Ile Asp Ser Asp Ser Glu 290
295 300Val Thr Ile Thr Tyr Glu305
310159681PRTAspergillus niger 159Met Arg Cys Ser Leu Ile Ser Leu
Leu Gly Leu Ala Ala Ile Pro Ala1 5 10
15Leu Gly Gly Cys Pro Phe Ala His Thr Ala Asn Met Gly Ile
Asp Asn 20 25 30Met Val Lys
Ala His Ala His Met Ser Arg Pro Leu Ile Ala Ser Lys 35
40 45Ser Ser Pro Ser Thr Val Pro Thr Ser Ser Ser
Thr Pro Ser Val Gly 50 55 60Gln Lys
Gly Val Phe Met Met Asn Arg Ile Ala Pro Gly Thr Ser Glu65
70 75 80Leu Tyr Ile Ala Asn Thr Asp
Gly Ser Asn Glu Arg Pro Leu Leu Ser 85 90
95Asn Pro Val Tyr Glu Tyr His Ala Ser Phe Ser Pro Asp
Val Glu Trp 100 105 110Ile Thr
Phe Thr Ser Glu Arg Asn Gly Asp Gly Asn Ser Asp Ile Tyr 115
120 125Arg Val Arg Thr Asn Gly Ser Asp Leu Gln
Glu Leu Val Ala Thr Pro 130 135 140Ala
Val Glu Asp Ser Val Val Ile Ser Pro Asn Gly Arg Leu Ala Ala145
150 155 160Tyr Val Ser Thr Ala Asn
Asn Met Lys Ala Asn Ile Trp Ile Leu Asp 165
170 175Leu Gln Thr Gly Ala Gln Trp Asn Leu Thr Asn Thr
Pro Thr Thr Ala 180 185 190Ala
Asn Ser Ser Leu Met Glu Ser Tyr Leu Arg Pro Ala Trp Ser Pro 195
200 205Asp Gly Glu Trp Ile Ala Phe Ser Ser
Asp Arg Asn Thr Gln Trp Asp 210 215
220Gly His Gly Val Pro Thr Phe Leu Gly Arg Thr Gly Trp Glu Thr Thr225
230 235 240Gln Glu Leu Ser
Leu Tyr Ala Ile Arg Pro Asn Gly Ser Asp Phe Arg 245
250 255Gln Ile Ile Ser Lys Pro Tyr Tyr Ser Leu
Gly Ser Pro Lys Trp Ser 260 265
270Ala Asp Gly Lys Arg Ile Val Tyr Tyr Glu Met Thr Arg Glu Asp Thr
275 280 285Tyr Asn Ala His Arg Pro Glu
Thr Ile Thr Thr Ala Asn Ser Thr Ile 290 295
300Met Ser Val Asp Phe Glu Thr Gly Thr Asp Val Arg Val Glu Val
Ala305 310 315 320Gly Ser
Gly Val Lys Gln Phe Pro Gln Tyr Leu Asp Lys Asn Gly Thr
325 330 335Ile Ala Tyr Thr Leu Lys Gly
Gly Thr Ser Glu Gly Phe Tyr Thr Thr 340 345
350Ala Gly Leu Tyr Val Asn Thr Thr Ser Ala Thr Leu Arg Ser
Pro Ala 355 360 365Trp Ser Pro Asp
Gly Lys Gln Val Val Tyr Glu Lys Ser Thr Trp Ser 370
375 380Ile Arg Ser Gly Tyr Lys Gln Leu Tyr Ser Trp Asp
Ser Asp Trp Asp385 390 395
400Tyr Arg Phe Thr Asp Val Phe Pro Gln Val Ser His Gln Glu Arg Val
405 410 415Ala Ile Thr Gln Lys
Gln Leu Gly Asn Ser Ser Ile Val Thr Leu Asn 420
425 430Thr Thr Gly Gly Asp Leu Gln Leu Val Tyr Asp Pro
Ser Thr Ala Asp 435 440 445Phe Val
Ser Asp Asp Glu Thr Thr Gly Leu Ser Ala Tyr Gln Pro Ser 450
455 460Trp Ser Pro Cys Gly Glu Trp Leu Val Phe Gly
Val Gly Phe Trp Phe465 470 475
480Glu Thr Arg Glu Ala Ser Gly Gly Trp Ile Val Arg Ala Thr Ala Asn
485 490 495Gly Ser Tyr Ser
Glu Val Leu Val Asn Ser Ser Tyr Ser Ile Thr Glu 500
505 510Asp Gly Ala Leu Asn Ser Gly Phe Pro Ser Phe
Ser Pro Asp Gly Lys 515 520 525Lys
Val Val Tyr Arg Val Trp Gly Ala Asp Thr Ala Thr Tyr Gly Asn 530
535 540Ala Ser Glu Ile Gly Leu Arg Val Leu Asp
Leu Glu Thr Arg Lys Thr545 550 555
560Thr Val Leu Thr Thr Glu Trp Asp Asn Leu Pro Gln Phe Ser Pro
Asp 565 570 575Gly Glu Leu
Ile Leu Phe Thr Arg Lys Thr Ser Thr Tyr Asn Tyr Asp 580
585 590Val Cys Thr Ile Arg Pro Asp Gly Thr Asp
Leu Arg Val Leu Thr Ser 595 600
605Ser Gly Ala Asn Asp Ala His Ala Val Trp Ser Gln Asp Gly Arg Ile 610
615 620Met Trp Ser Thr Gly Met Tyr Gly
Phe Arg Phe Glu Cys Ala Leu Tyr625 630
635 640Gly Asp Thr Phe Gln Pro Tyr Gly Gln Val Met Ile
Met Asp Ala Asp 645 650
655Gly Gly Asn Lys Lys Leu Met Thr Asn Ser Met Trp Glu Asp Ser Met
660 665 670Pro Leu Phe Leu Pro Arg
Glu Val Leu 675 680160624PRTAspergillus niger
160Met Pro Pro Asp Ala Lys Ser Pro Gly Tyr Gln Pro Gly Met Ala Val1
5 10 15Leu Pro Ser Arg Pro His
Pro Ala Lys Gly Lys Ala Ile Arg Phe Leu 20 25
30Leu Ser Leu Ala Leu Val Ala Phe Ala Ile Val Gln Leu
Cys Gly Asn 35 40 45Phe His Lys
Asn Arg Ser Val Glu Gln Gln Leu Gln Ser Gln Thr Leu 50
55 60Asp Asp Glu Ser Phe Lys Trp Glu Asp Val Thr Pro
Thr Lys Gln Leu65 70 75
80Val Tyr His Pro Cys Phe Gly Asp His Glu Cys Ala Arg Leu Ser Leu
85 90 95Pro Met Asn Trp Asn Arg
Thr Asp Gly Glu Gly Ser Lys Ile Ala Leu 100
105 110Ala Val Ile Lys Leu Pro Ala Lys Val Pro Val Thr
Asp Ala Arg Tyr 115 120 125Gly Gly
Ala Ile Leu Leu Asn Pro Gly Gly Pro Gly Gly Ser Gly Val 130
135 140Ser Met Val Phe Arg Tyr Gly Lys Ala Ile Gln
Thr Ile Val Asp Ser145 150 155
160Pro Glu Ser Pro Ser Ala Asp Ser Ala Ser Gly Lys Tyr Phe Asp Val
165 170 175Val Ser Phe Asp
Pro Arg Gly Val Asn Asn Thr Thr Pro Asn Phe Ser 180
185 190Cys Phe Pro Asp Pro Ala Thr Arg Lys Ala Trp
Leu Leu Gln Ser Glu 195 200 205Ala
Glu Gly Leu Leu Gly Ser Ser Glu Gly Val Phe Asp Thr Arg Trp 210
215 220Ala Arg Tyr Glu Ala Phe Glu Arg Leu Leu
Ser Thr Ala Pro Asn Thr225 230 235
240Phe Pro Val Gly Thr Asn Val Asp Ala Glu Arg Ile Arg Leu His
Asn 245 250 255Arg Trp Lys
Lys Gly Glu Glu Lys Leu Leu Tyr Trp Gly Phe Ser Tyr 260
265 270Gly Thr Ile Leu Gly Ser Thr Phe Ala Ala
Met Gln Pro His Arg Ile 275 280
285Asn Arg Ala Val Ile Asp Gly Val Cys Asn Ala Asp Asp Tyr Tyr Ala 290
295 300Gly Asn Trp Leu Thr Asn Leu Gln
Asp Ser Asp Ala Ala Phe Asn Lys305 310
315 320Phe Phe Glu Tyr Cys Tyr Thr Ala Gly Pro Ser Ala
Cys Pro Phe Ala 325 330
335Leu Gly Gly Asp Pro Glu Asp Leu Lys Ser Arg Tyr Glu Gln Ile Leu
340 345 350Thr Asn Leu Thr Ser Ser
Pro Ile Ala Val Ser Pro Ser Gly Asn Arg 355 360
365Gly Pro Glu Ile Ile Thr Tyr Ser Asp Val Lys Ser Leu Val
Val Gln 370 375 380Ala Leu Tyr Val Pro
Leu Lys Leu Phe Asp Leu Val Ala Arg Leu Leu385 390
395 400Ala Glu Leu Glu Gln Gly Asn Gly Ser Ser
Phe Ala Asp Leu Lys Tyr 405 410
415Glu Ala Lys Gln Trp Pro Val Pro Pro Pro Cys Asp Ser Ser Ser Thr
420 425 430Gln Tyr Lys Val Pro
Gly Glu Ser Asp Gln Glu Ala Gly Arg Asn Ile 435
440 445Leu Cys Thr Asp Gly Pro Gly Leu Asp Gly Thr Ala
Lys Glu Asp Phe 450 455 460Arg Ser Tyr
Trp Asn Met Leu Arg Gly Gln Ser Lys Ala Val Gly Asp465
470 475 480Phe Trp Ala Glu Val Arg Met
Ser Cys Val Lys Leu Glu Thr Arg Pro 485
490 495Glu Trp Arg Tyr Asp Gly Met Arg Ile Gln Gly Pro
Phe Ala Gly Asn 500 505 510Thr
Ser His Pro Leu Leu Phe Ile Gly Asn Thr Tyr Asp Pro Val Thr 515
520 525Pro Leu Arg Asn Ala His Thr Met Ala
Arg Gly Phe Pro Glu Ser Ile 530 535
540Val Leu Glu Gln Asn Ser Val Gly His Cys Thr Leu Ser Gly Pro Ser545
550 555 560Leu Cys Thr Ala
Lys Ala Ile Arg Gln Tyr Phe Gln Thr Gly Glu Leu 565
570 575Pro Asp Pro Gly Thr Val Cys Gln Val Glu
Glu Leu Pro Phe Arg Leu 580 585
590Ala Gly Tyr Glu Arg Ser Gln Val Met Ser Pro Gly Asp Thr Glu Leu
595 600 605Met Ser Ala Leu His Ser Leu
Ser Glu Phe Arg His Leu Leu Gly Ala 610 615
620161554PRTAspergillus niger 161Met Leu Ser Ser Leu Leu Leu Gly Gly
Leu Leu Gly Leu Ala Thr Ala1 5 10
15Gln Phe Pro Pro Glu Pro Glu Gly Ile Thr Val Leu Lys Ser Lys
Leu 20 25 30His Glu Asn Val
Thr Ile Ser Phe Lys Glu Pro Gly Ile Cys Glu Thr 35
40 45Thr Pro Gly Val Arg Ser Tyr Ser Gly Tyr Val His
Leu Pro Pro Ala 50 55 60Ser Thr Ser
Phe Phe Trp Phe Phe Glu Ala Arg Lys Asp Pro Ser Asn65 70
75 80Ala Pro Leu Ala Ile Trp Leu Asn
Gly Gly Pro Gly Gly Ser Ser Leu 85 90
95Met Gly Leu Leu Glu Glu Leu Gly Pro Cys Ser Ile Ala Ser
Asp Ser 100 105 110Lys Thr Thr
Val Leu Asn Pro Trp Ser Trp Asn Asn Glu Val Asn Leu 115
120 125Leu Phe Leu Asp Gln Pro Thr Gln Val Gly Phe
Ser Tyr Asp Val Pro 130 135 140Thr Asn
Gly Thr Leu Thr Ala Asn Gly Thr Ala Phe Ala Ala His Ala145
150 155 160Leu Trp His Phe Ala Gln Thr
Trp Phe Phe Glu Phe Pro His Tyr Lys 165
170 175Pro Asn Asp Asp Arg Val Ser Leu Trp Ala Glu Ser
Tyr Gly Gly His 180 185 190Tyr
Gly Pro Gly Ile Phe Arg Phe Phe Gln Gln Gln Asn Asp Lys Ile 195
200 205Ala Glu Gly Thr Ala Glu Asp Gly Ala
Gln Tyr Leu His Leu Asp Thr 210 215
220Leu Gly Ile Val Asn Gly Leu Met Asp Met Val Ile Gln Glu Glu Ala225
230 235 240Tyr Ile Thr Trp
Pro Tyr Asn Asn Val Arg Leu Ala Pro Ser Ser Phe 245
250 255Asn Ser Arg Gly Phe Arg Asp Gln Ala Leu
Ala Cys Glu Ala Ala Leu 260 265
270Lys Glu Arg Asp Ser Gly Leu Pro His Ser Gly Lys Asn Ile Ser Glu
275 280 285Ile Cys Gly Gly Leu Ala Leu
Glu Trp Gly Asp Gly Pro Ile Thr Tyr 290 295
300Tyr His Thr Phe Asn Arg Gly Trp Tyr Asp Ile Ala His Pro Lys
Asn305 310 315 320Asp Pro
Phe Pro Ala Lys His Met Leu Gly Tyr Leu Thr Gln Glu Ser
325 330 335Val Leu Ala Ala Leu Gly Val
Pro Val Asn Phe Thr Ser Ser Ser Ser 340 345
350Ala Val Ala Thr Gln Phe Ile Lys Thr Phe Asp Ile Val His
Gly Gly 355 360 365Phe Leu Asp Ala
Ile Gly Tyr Leu Leu Asp Ser Gly Val Lys Val His 370
375 380Met Met Tyr Gly Asp Arg Asp Tyr Ala Cys Asn Trp
Val Gly Gly Glu385 390 395
400Lys Ala Ser Leu Ala Val Pro Tyr Ser Arg Ile Thr Glu Phe Ala Asp
405 410 415Thr Gly Tyr Ser Pro
Leu Leu Thr Pro Asp Gly Ile Ser Gly Met Thr 420
425 430Arg Gln Leu Gly Asn Tyr Ser Phe Thr Arg Val Phe
Gln Ala Gly His 435 440 445Glu Val
Pro Ser Tyr Gln Pro Val Ala Ala Tyr Glu Ile Phe Met Arg 450
455 460Ala Thr Phe Asn Lys Asp Ile Pro Thr Gly Leu
Leu Ala Val Asp Asp465 470 475
480Glu Phe Gln Ser Val Gly Pro Lys Asp Thr Trp His Ile Lys Asn Ile
485 490 495Pro Pro Ile Met
Pro Lys Pro Gln Cys Tyr Val Leu Ser Pro Gly Thr 500
505 510Cys Thr Pro Glu Val Trp Glu Thr Val Leu Asn
Gly Ser Ala Thr Val 515 520 525Lys
Asp Trp Tyr Val Val Asp Asp Ser Ala Gly Val Glu Asp His Glu 530
535 540Gly Phe Ser Ile Leu Gly Gly Asp Glu
Leu545 550162578PRTAspergillus niger 162Met Thr Arg Phe
Gln Leu Leu Pro Leu Val Ala Gly Leu Leu Ala Pro1 5
10 15Ser Ile Ala Ala Leu Ser Ile Pro Ser Pro
Gln Gln Ile Leu Asp Ser 20 25
30Leu Thr Phe Gly Glu His Thr Asp Gly Phe Cys Pro Leu Ala Pro Lys
35 40 45Val Glu Val Pro Asp Asp Gly Phe
Phe Pro Ala Leu Lys Phe Val Glu 50 55
60Asp Ala Ser Phe Lys Ser Arg Gln Val Asn Arg Leu Ser Arg Ala Val65
70 75 80Gln Val Pro Thr Ala
Ile Asp Asp Tyr Met Lys Asp Pro Tyr Asp Glu 85
90 95Lys Phe Ala Pro Phe Leu Asp Phe Gln Lys Leu
Leu Gln Thr Leu Phe 100 105
110Pro Leu Thr His Ser Tyr Ala Arg Val Asp His Ile Asn Arg Phe Gly
115 120 125Leu Val Phe Thr Leu Asn Gly
Thr Asp Asp Ser Leu Lys Pro Leu Leu 130 135
140Phe Thr Ala His Gln Asp Val Val Pro Ile Asn Asp Pro Ala Asp
Trp145 150 155 160Thr Tyr
Pro Pro Phe Asp Gly His Tyr Asp Gly Glu Trp Leu Trp Gly
165 170 175Arg Gly Ala Ser Asp Cys Lys
Asn Val Leu Ile Gly Leu Met Ser Val 180 185
190Val Glu Asp Leu Leu Ser Gln Lys Trp Glu Pro Thr Arg Thr
Val Val 195 200 205Leu Ala Phe Gly
Phe Asp Glu Glu Ser His Gly Phe Leu Gly Ala Gly 210
215 220Ser Ile Ala Lys Phe Leu Glu Lys Lys Tyr Gly Pro
Asp Ser Phe Glu225 230 235
240Phe Ile Leu Asp Glu Gly Gly Met Gly Leu Glu Val Leu Asp Asp Asn
245 250 255Asn Asn Gly Val Val
Tyr Ala Leu Pro Gly Val Gly Glu Lys Gly Ser 260
265 270Ile Asp Val Val Leu Thr Leu Ala Val Pro Gly Gly
His Ser Ser Val 275 280 285Pro Pro
Pro His Thr Gly Ile Gly Ile Ile Ala Glu Ile Ile Tyr Glu 290
295 300Leu Glu Arg Gln Asp Leu Phe Val Pro Val Leu
Asp Thr His His Pro305 310 315
320Thr Arg Lys Met Leu Glu Cys Gln Val Arg His Ser Pro Ser Gln Val
325 330 335Glu Pro Trp Leu
Ala Ser Ala Leu Gln Ser Ser Asp Tyr Ile Ser Leu 340
345 350Ala Glu Lys Leu Ala Ser Ser Arg Gly Asp Lys
Phe Arg Phe Ile Leu 355 360 365Gln
Thr Ser Gln Ala Ala Asp Ile Ile Asn Gly Gly Val Lys Ser Asn 370
375 380Ala Leu Pro Glu Lys Ile Asn Ala Leu Val
Asn Tyr Arg Ile Ala Leu385 390 395
400His Gln Thr Pro Asp Asp Ile Lys Asn Arg Ala Val Glu Ile Ile
Ser 405 410 415Pro Ile Val
Lys Lys Tyr Asn Leu Ser Leu Thr Ala Phe Pro Glu Ser 420
425 430Asp Thr Val Asp Pro Ser Leu Asn Asn His
Leu Thr Leu Thr Thr Leu 435 440
445Ser Gly Ala Leu Ser Pro Ala Pro Val Ser Pro Thr Asp Ile Asp Thr 450
455 460Asp Ala Val Trp Ala Arg Phe Ser
Gly Val Thr Arg Ser Val Phe Glu465 470
475 480Ser Val Pro Ser Leu Glu Gly Arg Lys Val Val Val
Ser Gly Asp Ile 485 490
495Met Thr Gly Asn Thr Asp Thr Arg Phe Tyr Trp Ala Leu Ser Arg Asn
500 505 510Ile Tyr Arg Trp Ser Pro
Ser Arg Ala Gly Lys Ala Leu Asn Ile His 515 520
525Thr Val Asp Glu Arg Ile Asp Ile Asp Ile His Leu Glu Ala
Met Met 530 535 540Leu Tyr Tyr Asp Leu
Ile Arg Ser Phe Asp Gly Arg Thr Asp Ser Ser545 550
555 560Val Ile Ser Ala Ala Ser Ala Ala Ala Asp
Asp Glu Leu Ala His Asp 565 570
575Val Leu163456PRTAspergillus niger 163Met Lys Ser Thr Thr Leu Leu
Ser Leu Ala Trp Ala Ala Gln Ser Ala1 5 10
15Tyr Ser Leu Ser Ile His Glu Arg Asp Glu Pro Ala Thr
Leu Gln Phe 20 25 30Asn Phe
Glu Arg Arg Gln Ile Ala Asp Arg Ser Arg Arg Lys Arg Ser 35
40 45Thr Ala Ser Ala Asp Leu Val Asn Leu Ala
Thr Asn Leu Gly Tyr Thr 50 55 60Met
Asn Leu Thr Leu Gly Thr Pro Gly Gln Glu Val Ser Val Thr Leu65
70 75 80Asp Thr Gly Ser Ser Asp
Leu Trp Val Asn Gly Ala Asn Ser Ser Val 85
90 95Cys Pro Cys Thr Asp Tyr Gly Ser Tyr Asn Ser Ser
Ala Ser Ser Thr 100 105 110Tyr
Thr Phe Val Asn Asp Glu Phe Tyr Ile Gln Tyr Val Asp Gly Ser 115
120 125Glu Ala Thr Gly Asp Tyr Val Asn Asp
Thr Leu Lys Phe Ser Asn Val 130 135
140Thr Leu Thr Asn Phe Gln Phe Ala Val Ala Tyr Asp Gly Asp Ser Glu145
150 155 160Glu Gly Val Leu
Gly Ile Gly Tyr Ala Ser Asn Glu Ala Ser Gln Ala 165
170 175Thr Val Gly Gly Gly Glu Tyr Thr Asn Phe
Pro Glu Ala Leu Val Asp 180 185
190Gln Gly Ala Ile Asn Trp Pro Ala Tyr Ser Leu Trp Leu Asp Asp Leu
195 200 205Asp Glu Gly Lys Gly Thr Ile
Leu Phe Gly Gly Val Asn Thr Ala Lys 210 215
220Tyr Tyr Gly Ser Leu Gln Thr Leu Pro Ile Val Ser Ile Glu Asp
Met225 230 235 240Tyr Val
Glu Phe Ala Val Asn Leu Thr Ala Val His Leu Glu Lys Asn
245 250 255Gly Asn Ser Val Ser Val Asn
Asn Ser Ala Thr Gln Phe Pro Ile Pro 260 265
270Ala Val Leu Asp Ser Gly Thr Ala Leu Thr Tyr Ile Pro Thr
Ser Ala 275 280 285Ala Ala Ser Ile
Tyr Glu Ala Val Gly Ala Gln Tyr Leu Ser Glu Tyr 290
295 300Gly Tyr Gly Val Ile Glu Cys Asp Val Lys Asp Glu
Asp Phe Thr Phe305 310 315
320Leu Phe Asp Phe Gly Ser Phe Asn Met Ser Val Asp Ile Ser Glu Met
325 330 335Ile Leu Glu Ala Ser
Ser Asp Met Thr Asp Met Asn Val Cys Thr Phe 340
345 350Gly Leu Ala Val Ile Glu Asn Glu Ala Leu Leu Gly
Asp Thr Phe Leu 355 360 365Arg Ser
Ala Tyr Val Val Tyr Asp Leu Gly Asn Asn Glu Ile Ser Leu 370
375 380Ala Lys Ala Asn Phe Asn Pro Gly Glu Asp His
Val Leu Glu Ile Gly385 390 395
400Thr Gly Ser Asp Ala Val Pro Lys Ala Thr Gly Ala Thr Ala Thr Gly
405 410 415Ala Ala Ala Thr
Ser Thr Ala Ser Ser Asp Lys Ser Asp Lys Glu Ser 420
425 430Ser Ala Thr Val Pro Arg Ser Gln Ile Val Ser
Leu Val Ala Gly Val 435 440 445Leu
Val Gly Val Phe Leu Val Leu 450
455164664PRTAspergillus niger 164Met Leu Val Arg Gln Leu Ala Leu Ala Leu
Ala Ile Ala Ala Leu Ser1 5 10
15Asp Ala Ile Pro Thr Ser Ile Lys His Val Leu His Glu Lys Arg His
20 25 30Lys Pro Ala Ser Asp Trp
Val Lys Gly Ala Arg Val Glu Ser Asp Ala 35 40
45Val Leu Pro Met Arg Ile Gly Leu Ala Gln Asn Asn Leu Asp
Lys Gly 50 55 60Tyr Asp Phe Leu Met
Glu Val Ser Asp Pro Lys Ser Ser Lys Tyr Gly65 70
75 80Gln Tyr Trp Ser Ala Asp Glu Val His Asp
Ile Phe Ser Pro Ser Glu 85 90
95Glu Ala Val Glu Ala Val Arg Glu Trp Leu Val Ala Ser Gly Ile His
100 105 110Pro Ser Arg Val Val
His Ser Asp Asn Lys Gly Trp Leu Ala Phe Asp 115
120 125Ala Tyr Ala His Glu Ala Glu Arg Leu Phe Met Thr
Glu Phe His Glu 130 135 140His Glu Ser
Asp Arg Ser Ala Lys Ile Arg Val Gly Cys Asp Gln Tyr145
150 155 160His Val Pro Glu His Ile Gln
Lys His Ile Asp Tyr Ile Thr Pro Gly 165
170 175Val Lys Leu Thr Gln Val Val Lys Arg Thr Asn Lys
Val Lys Arg Ala 180 185 190Ser
Gln Leu Ala His Ser Ser Lys Ala Lys Ser Ala Ala Gln Gly Pro 195
200 205Gln Pro Leu Pro Asn Lys Ala Lys Phe
Leu Pro Glu Asp Leu Arg Gly 210 215
220Cys Gly Tyr Asn Ile Thr Pro Ser Cys Ile Lys Ala Leu Tyr Gln Ile225
230 235 240Pro Asp Ala Lys
Thr Ala Thr Pro Asn Asn Ser Leu Gly Leu Tyr Glu 245
250 255Gln Gly Asp Tyr Phe Ala Lys Ser Asp Leu
Asp Leu Phe Tyr Lys Glu 260 265
270Tyr Ala Pro Trp Val Pro Gln Gly Thr Tyr Pro Ile Pro Ala Leu Ile
275 280 285Asp Gly Ala Asn Tyr Ser Val
Pro Ser Tyr Ser Ser Leu Asn Thr Gly 290 295
300Glu Ser Asp Ile Asp Ile Asp Met Ala Tyr Ser Leu Leu Tyr Pro
Gln305 310 315 320Gln Val
Thr Leu Tyr Gln Val Asp Asp Gln Leu Tyr Glu Pro Val Glu
325 330 335Val Asp Thr Thr Asn Leu Phe
Asn Thr Phe Leu Asp Ala Leu Asp Gly 340 345
350Ser Tyr Cys Thr Tyr Ser Ala Tyr Gly Glu Thr Gly Asp Asp
Pro Ser 355 360 365Ile Asp Pro Val
Tyr Pro Asp Thr Arg Pro Gly Gly Tyr Lys Gly Lys 370
375 380Leu Gln Cys Gly Val Tyr Lys Pro Thr Asn Val Ile
Ser Ala Ser Tyr385 390 395
400Gly Gln Ser Glu Ala Asp Leu Pro Val Ser Tyr Thr Lys Arg Gln Cys
405 410 415Asn Glu Phe Met Lys
Leu Gly Leu Gln Gly His Ser Ile Leu Phe Ala 420
425 430Ser Gly Asp Tyr Gly Val Ala Ser Phe Ala Gly Asp
Gly Asp Glu Asn 435 440 445Gly Cys
Leu Gly Pro Glu Gly Lys Ile Phe Asn Pro Gln Tyr Pro Ser 450
455 460Asn Cys Pro Tyr Val Thr Ser Val Gly Gly Thr
Met Leu Tyr Gly Tyr465 470 475
480Gln Thr Val Asn Asp Ser Glu Ser Val Met His Val Asn Leu Gly Gly
485 490 495Thr Ala Ser Asn
Phe Ser Thr Ser Gly Gly Phe Ser Asn Tyr Phe Pro 500
505 510Gln Pro Ala Tyr Gln Phe Ala Ala Val Glu Gln
Tyr Phe Gln Ser Ala 515 520 525Asn
Leu Ser Tyr Pro Tyr Tyr Ser Glu Phe Glu Val Asp Val Asn Thr 530
535 540Thr Lys Gly Leu Tyr Asn Arg Leu Gly Arg
Ala Tyr Pro Asp Val Ser545 550 555
560Ala Asn Gly Ala His Phe Arg Ala Tyr Met Asp Gly Tyr Asp Tyr
His 565 570 575Trp Tyr Gly
Ser Ser Leu Ala Ser Pro Leu Phe Ala Ser Val Leu Thr 580
585 590Leu Leu Asn Glu Glu Arg Phe Ala Ile Gly
Lys Gly Pro Val Gly Phe 595 600
605Val Asn Pro Val Leu Tyr Ala Tyr Pro Gln Val Leu Asn Asp Ile Thr 610
615 620Asn Gly Thr Asn Ala Gly Cys Gly
Thr Tyr Gly Phe Ser Ala Ile Glu625 630
635 640Gly Trp Asp Pro Ala Ser Gly Leu Gly Thr Pro Asn
Tyr Pro Leu Met 645 650
655Lys Glu Leu Phe Leu Ser Leu Pro 660165520PRTAspergillus
niger 165Met Arg Val Thr Thr Ala Ile Ala Ser Leu Leu Leu Val Gly Ser Ala1
5 10 15Thr Ser Leu Gln
Asn Pro His Arg Arg Ala Val Pro Pro Pro Leu Ser 20
25 30His Arg Ser Val Ala Ser Arg Ser Val Pro Val
Glu Arg Arg Thr Thr 35 40 45Asp
Phe Glu Tyr Leu Thr Asn Lys Thr Ala Arg Phe Leu Val Asn Gly 50
55 60Thr Ser Ile Pro Glu Val Asp Phe Asp Val
Gly Glu Ser Tyr Ala Gly65 70 75
80Leu Leu Pro Asn Thr Pro Thr Gly Asn Ser Ser Leu Phe Phe Trp
Phe 85 90 95Phe Pro Ser
Gln Asn Pro Glu Ala Ser Asp Glu Ile Thr Ile Trp Leu 100
105 110Asn Gly Gly Pro Gly Cys Ser Ser Leu Asp
Gly Leu Leu Gln Glu Asn 115 120
125Gly Pro Phe Leu Trp Gln Pro Gly Thr Tyr Lys Pro Val Pro Asn Pro 130
135 140Tyr Ser Trp Thr Asn Leu Thr Asn
Val Val Tyr Ile Asp Gln Pro Ala145 150
155 160Gly Thr Gly Phe Ser Pro Gly Pro Ser Thr Val Asn
Asn Glu Glu Asp 165 170
175Val Ala Ala Gln Phe Asn Ser Trp Phe Lys His Phe Val Asp Thr Phe
180 185 190Asp Leu His Gly Arg Lys
Val Tyr Ile Thr Gly Glu Ser Tyr Ala Gly 195 200
205Met Tyr Val Pro Tyr Ile Ala Asp Ala Met Leu Asn Glu Glu
Asp Thr 210 215 220Thr Tyr Phe Asn Leu
Lys Gly Ile Gln Ile Asn Asp Pro Ser Ile Asn225 230
235 240Ser Asp Ser Val Met Met Tyr Ser Pro Ala
Val Arg His Leu Asn His 245 250
255Tyr Asn Asn Ile Phe Gln Leu Asn Ser Thr Phe Leu Ser Tyr Ile Asn
260 265 270Ala Lys Ala Asp Lys
Cys Gly Tyr Asn Ala Phe Leu Asp Lys Ala Ile 275
280 285Thr Tyr Pro Pro Pro Ser Pro Phe Pro Thr Ala Pro
Glu Ile Thr Glu 290 295 300Asp Cys Gln
Val Trp Asp Glu Val Val Met Ala Ala Tyr Asp Ile Asn305
310 315 320Pro Cys Phe Asn Tyr Tyr His
Leu Ile Asp Phe Cys Pro Tyr Leu Trp 325
330 335Asp Val Leu Gly Phe Pro Ser Leu Ala Ser Gly Pro
Asn Asn Tyr Phe 340 345 350Asn
Arg Ser Asp Val Gln Lys Ile Leu His Val Pro Pro Thr Asp Tyr 355
360 365Ser Val Cys Ser Glu Thr Val Ile Phe
Ala Asn Gly Asp Gly Ser Asp 370 375
380Pro Ser Ser Trp Gly Pro Leu Pro Ser Val Ile Glu Arg Thr Asn Asn385
390 395 400Thr Ile Ile Gly
His Gly Trp Leu Asp Tyr Leu Leu Phe Leu Asn Gly 405
410 415Ser Leu Ala Thr Ile Gln Asn Met Thr Trp
Asn Gly Lys Gln Gly Phe 420 425
430Gln Arg Pro Pro Val Glu Pro Leu Phe Val Pro Tyr His Tyr Gly Leu
435 440 445Ala Glu Leu Tyr Trp Gly Asp
Glu Pro Asp Pro Tyr Asn Leu Asp Ala 450 455
460Gly Ala Gly Tyr Leu Gly Thr Ala His Thr Glu Arg Gly Leu Thr
Phe465 470 475 480Ser Ser
Val Tyr Leu Ser Gly His Glu Ile Pro Gln Tyr Val Pro Gly
485 490 495Ala Ala Tyr Arg Gln Leu Glu
Phe Leu Leu Gly Arg Ile Ser Ser Leu 500 505
510Ser Ala Lys Gly Asn Tyr Thr Ser 515
520166551PRTAspergillus niger 166Met Arg Gly Ser Arg Leu Val Leu Leu Leu
Pro Leu Ala Ala Leu Ser1 5 10
15Cys Ala Met Pro Glu Asn Glu Trp Ser Ser Thr Ile Arg Arg Gln Leu
20 25 30Pro Lys Ala Ser Thr Gly
Val Lys Ser Ile Lys Thr Pro Asn Asn Val 35 40
45Thr Ile Arg Tyr Lys Glu Pro Gly Thr Glu Gly Ile Cys Glu
Thr Thr 50 55 60Pro Gly Val Lys Ser
Tyr Ser Gly Tyr Val Asp Leu Ser Pro Glu Ser65 70
75 80His Thr Phe Phe Trp Phe Phe Glu Ser Arg
Arg Asp Pro Glu Asn Asp 85 90
95Pro Val Thr Leu Trp Leu Asn Gly Gly Pro Gly Ser Asp Ser Leu Ile
100 105 110Gly Leu Phe Glu Glu
Leu Gly Pro Cys His Ile Thr Pro Glu Tyr Glu 115
120 125Ser Ile Ile Asn Gln Tyr Ser Trp Asn Glu Val Thr
Asn Leu Leu Phe 130 135 140Leu Ser Gln
Pro Leu Gly Val Gly Phe Ser Tyr Ser Glu Thr Glu Ala145
150 155 160Gly Ser Leu Asn Pro Phe Thr
Gly Ala Val Glu Asn Ala Ser Phe Ala 165
170 175Gly Val Gln Gly Arg Tyr Pro Val Ile Asp Ala Thr
Ile Ile Asp Thr 180 185 190Thr
Asp Ile Ala Ala Arg Ala Thr Trp Glu Val Leu Gln Gly Phe Leu 195
200 205Ser Gly Leu Ser Gln Leu Asp Ser Glu
Val Lys Ser Lys Glu Phe Asn 210 215
220Leu Trp Thr Glu Ser Tyr Gly Gly His Tyr Gly Pro Ala Phe Phe Asn225
230 235 240His Phe Tyr Glu
Gln Asn Ser Lys Ile Ala Ser Gly Glu Val Asn Gly 245
250 255Val Gln Leu Asn Phe Asn Ser Leu Gly Ile
Ile Asn Gly Ile Ile Asp 260 265
270Ala Ala Ile Gln Ala Asp Tyr Tyr Ala Asp Phe Ala Val Asn Asn Thr
275 280 285Tyr Gly Ile Lys Ala Val Asn
Asp Thr Val Tyr Asn Tyr Met Lys Phe 290 295
300Ala Asn Thr Met Pro Asn Gly Cys Gln Asp Gln Val Ala Ser Cys
Lys305 310 315 320Leu Thr
Asn Arg Thr Ser Leu Ser Asp Tyr Ala Ile Cys Thr Glu Ala
325 330 335Ala Asn Met Cys Arg Asp Asn
Val Glu Gly Pro Tyr Tyr Gln Phe Gly 340 345
350Gly Arg Gly Val Tyr Asp Ile Arg His Pro Tyr Asn Asp Pro
Thr Pro 355 360 365Pro Ser Tyr Phe
Val Asp Tyr Leu Lys Lys Asp Ser Val Met Asp Ala 370
375 380Ile Gly Val Asp Ile Asn Tyr Thr Glu Ser Ser Gly
Glu Val Tyr Tyr385 390 395
400Ala Phe Gln Gln Thr Gly Asp Phe Val Trp Pro Asn Phe Ile Glu Asp
405 410 415Leu Glu Glu Ile Leu
Gln Leu Pro Val Arg Val Ser Leu Ile Tyr Gly 420
425 430Asp Ala Asp Tyr Ile Cys Asn Trp Phe Gly Gly Gln
Ala Ile Ser Leu 435 440 445Ala Val
Asn Tyr Pro His Ala Ala Gln Phe Arg Ala Ala Gly Tyr Thr 450
455 460Pro Met Thr Val Asp Gly Val Glu Tyr Gly Glu
Thr Arg Glu Tyr Gly465 470 475
480Asn Phe Ser Phe Thr Arg Val Tyr Gln Ala Gly His Glu Val Pro Tyr
485 490 495Tyr Gln Pro Ile
Ala Ala Leu Gln Leu Phe Asn Arg Thr Leu Phe Gly 500
505 510Trp Asp Ile Ala Ala Gly Thr Thr Gln Ile Trp
Pro Glu Tyr Ser Thr 515 520 525Asn
Gly Thr Ser Gln Ala Thr His Thr Glu Ser Phe Val Pro Leu Ser 530
535 540Thr Ala Ser Ser Thr Val Asn545
550167623PRTAspergillus niger 167Met Pro Phe Pro Phe Ser Ser Ala Leu
Leu Gly Tyr Ile Leu Thr Thr1 5 10
15Ser Thr Thr Leu Thr Ser Leu Val Ala Gly Gln Tyr Tyr Pro Pro
Thr 20 25 30Pro Glu Asp Leu
Thr Val Ile His Ser Glu Ile Phe Pro Gly Ala Arg 35
40 45Ile Ser Tyr Lys Gln Pro Leu Gly Ile Cys Thr Thr
Thr Pro Ser Thr 50 55 60Pro Ser Tyr
Ser Gly Tyr Ile His Leu Pro Pro His Thr Leu Thr Asn65 70
75 80Leu Ser Ile Pro Gly Ile Ser Ile
Ser Gln Pro Tyr Pro Ile Asn Thr 85 90
95Phe Phe Trp Tyr Phe Pro Ser Arg His His His Asn Asn Asp
Thr Ser 100 105 110Pro Leu Thr
Ile Trp Met Asn Gly Gly Pro Gly Gly Ser Ser Met Ile 115
120 125Gly Leu Phe Gln Glu Asn Gly Pro Cys Thr Val
Asn Thr Asp Ser Asn 130 135 140Ser Thr
Ala Tyr Asn Pro Trp Ser Trp Asn Glu Tyr Val Asp Met Leu145
150 155 160Tyr Ile Glu Gln Pro Val Gln
Thr Gly Phe Ser Tyr Asp Val Leu Arg 165
170 175Asn Gly Thr Leu Asp Leu Asn Glu Thr Phe Leu Val
Gly Thr Leu Pro 180 185 190Ser
Gln Asp Val His Gly Thr Val Asn Gly Thr Val Asn Gly Gly Arg 195
200 205Ala Leu Trp Val Ala Leu Gln Val Trp
Leu Gly Glu Phe Ser Glu Tyr 210 215
220Val Ser Ser Val Asp Gly Asn Gly Gly Gly Asp Asp Arg Val Ser Ile225
230 235 240Trp Thr Glu Ser
Tyr Gly Gly Arg Tyr Gly Pro Ala Tyr Thr Ala Leu 245
250 255Phe Gln Glu Met Asn Glu Arg Ile Glu Ser
Gly Glu Val Ser Thr Gly 260 265
270Lys Lys Ile His Leu Asp Thr Leu Gly Ile Ile Asn Gly Cys Val Asp
275 280 285Leu Leu Val Gln Val Pro Ser
Phe Pro Glu Gln Ala Tyr Asn Asn Thr 290 295
300Tyr Gly Ile Glu Gly Ile Asn Arg Thr Leu Tyr Asp Arg Ala Met
Asp305 310 315 320Ser Trp
Ser Lys Pro Gly Gly Cys Arg Asp Met Ile Ile Glu Cys Arg
325 330 335Asp Ala Gly Glu Leu Gly Asp
Pro Leu Ile Ile Cys Glu Glu Ala Ser 340 345
350Asp Tyr Cys Ser Arg Glu Ile Lys Ser Leu Tyr Thr Asn Thr
Ser Gly 355 360 365Arg Gly Tyr Tyr
Asp Ile Ala His Phe Thr Pro Asp Ala Ala Leu Val 370
375 380Pro Tyr Phe Val Gly Phe Leu Asn Arg Pro Trp Val
Gln Lys Ala Leu385 390 395
400Gly Val Pro Val Asn Tyr Thr Met Ser Ser Glu Ala Val Gly Asn Ser
405 410 415Phe Ala Ser Thr Gly
Asp Tyr Pro Arg Asn Asp Pro Arg Gly Met Ile 420
425 430Gly Asp Ile Gly Tyr Leu Leu Asp Ser Gly Val Lys
Val Ala Met Val 435 440 445Tyr Gly
Asp Arg Asp Tyr Ala Cys Pro Trp Arg Gly Gly Glu Asp Val 450
455 460Ser Leu Leu Val Glu Tyr Glu Asp Ala Glu Lys
Phe Arg Ala Ala Gly465 470 475
480Tyr Ala Glu Val Gln Thr Lys Ser Ser Tyr Val Gly Gly Leu Val Arg
485 490 495Gln Tyr Gly Asn
Phe Ser Phe Thr Arg Val Phe Gln Ala Gly His Glu 500
505 510Val Pro Phe Tyr Gln Pro Glu Thr Ala Tyr Glu
Ile Phe Asn Arg Ala 515 520 525Gln
Phe Asn Trp Asp Ile Ala Thr Gly Gly Ile Ser Leu Glu Gln Asn 530
535 540Gln Ser Tyr Gly Thr Glu Gly Pro Ser Ser
Thr Trp His Ile Lys Asn545 550 555
560Glu Val Pro Glu Ser Pro Glu Pro Thr Cys Tyr Leu Leu Ala Met
Asp 565 570 575Ser Thr Cys
Thr Asp Glu Gln Arg Glu Arg Val Leu Ser Gly Asp Ala 580
585 590Val Val Arg Asp Trp Val Val Val Asp Asp
Ile Glu Ala Glu Ser Ser 595 600
605Phe Ser Gly Val Gly Asp Gln Leu Ala Gln Val Pro Leu Gly His 610
615 620168439PRTAspergillus niger 168Met Arg
Thr Ser Thr Leu Leu Leu Leu Trp Ser Thr Ala Gly Ala Ala1 5
10 15Leu Ala Ser Pro Tyr Pro Leu Pro
Asp Ser Gln Val Val Phe Ala Ala 20 25
30Asp His Glu Val Pro Asn Thr Gln Gly Lys His Val Val Asp Glu
Ala 35 40 45Ile Leu Ser Ala Leu
Asn Ala His Ser Asp Pro Val Ala Ala Met Val 50 55
60Ser Leu Arg Pro Glu Thr Ala Ala Phe Leu Ala Glu Pro Arg
Leu Leu65 70 75 80His
Ile Arg Gly Glu Glu Lys Ala Glu Trp Met Thr Glu Gly Asp Lys
85 90 95Leu Arg Leu Arg Gln Arg Gly
Lys Lys Phe Met Asp Ile Thr Glu His 100 105
110Gln Asp Phe Tyr Ala Glu Gln Ala Met Ala Ser Phe Ala Gly
Asp Pro 115 120 125Asn Leu Pro Lys
Leu Ser His Lys Gly Leu Val Lys Pro Leu Phe Ser 130
135 140Gln Ile Glu Thr Glu Arg Met His Asp Ile Leu Gln
His Met Thr Ser145 150 155
160Tyr Tyr Asn Arg Tyr Tyr Gly Asp Tyr His Gly Glu Met Ser Ser Glu
165 170 175Trp Leu His Asp Tyr
Ile Ala Ala Ile Ile Ser Lys Ser Pro Phe Arg 180
185 190Thr His Ile Ser Leu Glu Tyr Phe Thr His Pro Phe
Arg Gln Ser Ser 195 200 205Ile Ile
Ala Arg Phe Glu Pro Lys Val Arg Ser Phe Ser Gln Pro Leu 210
215 220Thr Ile Ile Gly Ala His Gln Asp Ser Ala Asn
Tyr Leu Phe Pro Leu225 230 235
240Leu Pro Ala Pro Gly Ala Asp Asp Asp Cys Ser Gly Thr Val Ser Ile
245 250 255Leu Glu Ala Phe
Arg Val Leu Ala Glu Asn Gly Tyr Thr Pro Lys Asp 260
265 270Gly Pro Val Glu Phe His Trp Tyr Ala Ala Glu
Glu Ala Gly Leu Leu 275 280 285Gly
Ser Gln Ala Ile Ala Arg Tyr Lys Lys Glu Gln Gly Ala Lys Ile 290
295 300Asp Ala Met Met Glu Phe Asp Met Thr Ala
Phe Ile Ala Arg Asn Ala305 310 315
320Thr Glu Thr Ile Gly Phe Val Ala Thr Gln Ala Asp Ala Ala Leu
Thr 325 330 335Asn Trp Ala
Leu Asn Leu Ser Arg Glu Tyr Ile Ser Ile Pro Ala Glu 340
345 350Val Tyr Glu Leu Gly Pro Asn Ala Gly Ser
Asp Tyr Met Ser Tyr Thr 355 360
365Lys Leu Asn Tyr Pro Ala Ala Phe Ala Ser Glu Gly Asn Pro Leu Ala 370
375 380Gly Gly Ser Phe Pro Gly Glu Met
Asp Pro Tyr Val His Gly Ile Lys385 390
395 400Asp Arg Met Asp Val Asp Asp Glu Thr Gly Val Phe
Ser Ile Glu His 405 410
415Met Ala Arg Phe Ser Glu Leu Ala Ile Ala Phe Val Val Glu Gln Ala
420 425 430Gly Trp Asp Asn Thr Trp
Arg 435169526PRTAspergillus niger 169Met Arg Ser Phe Ser Val Val
Ala Ala Ala Ser Leu Ala Leu Ser Trp1 5 10
15Ala Ser Leu Ala Gln Ala Ala Arg Pro Arg Leu Val Pro
Lys Pro Ile 20 25 30Ser Arg
Pro Ala Ser Ser Lys Ser Ala Ala Thr Thr Gly Glu Ala Tyr 35
40 45Phe Glu Gln Leu Leu Asp His His Asn Pro
Glu Lys Gly Thr Phe Ser 50 55 60Gln
Arg Tyr Trp Trp Ser Thr Glu Tyr Trp Gly Gly Pro Gly Ser Pro65
70 75 80Val Val Leu Phe Asn Pro
Gly Glu Val Ser Ala Asp Gly Tyr Glu Gly 85
90 95Tyr Leu Thr Asn Asp Thr Leu Thr Gly Val Tyr Ala
Gln Glu Ile Gln 100 105 110Gly
Ala Val Ile Leu Ile Glu His Arg Tyr Trp Gly Asp Ser Ser Pro 115
120 125Tyr Glu Val Leu Asn Ala Glu Thr Leu
Gln Tyr Leu Thr Leu Asp Gln 130 135
140Ser Ile Leu Asp Met Thr Tyr Phe Ala Glu Thr Val Lys Leu Gln Phe145
150 155 160Asp Asn Ser Ser
Arg Ser Asn Ala Gln Asn Ala Pro Trp Val Met Val 165
170 175Gly Gly Ser Tyr Ser Gly Ala Leu Thr Ala
Trp Thr Glu Ser Ile Ala 180 185
190Pro Gly Thr Phe Trp Ala Tyr His Ala Thr Ser Ala Pro Val Glu Ala
195 200 205Ile Tyr Asp Phe Trp Gln Tyr
Phe Tyr Pro Ile Gln Gln Gly Met Ala 210 215
220Gln Asn Cys Ser Lys Asp Val Ser Leu Val Ala Glu Tyr Val Asp
Lys225 230 235 240Ile Gly
Lys Asn Gly Thr Ala Lys Glu Gln Gln Glu Leu Lys Glu Leu
245 250 255Phe Gly Leu Gly Ala Val Glu
His Tyr Asp Asp Phe Ala Ala Val Leu 260 265
270Pro Asn Gly Pro Tyr Leu Trp Gln Asp Asn Asp Phe Val Thr
Gly Tyr 275 280 285Ser Ser Phe Phe
Gln Phe Cys Asp Ala Val Glu Gly Val Glu Ala Gly 290
295 300Ala Ala Val Thr Pro Gly Pro Glu Gly Val Gly Leu
Glu Lys Ala Leu305 310 315
320Ala Asn Tyr Ala Asn Trp Phe Asn Ser Thr Ile Leu Pro Asn Tyr Cys
325 330 335Ala Ser Tyr Gly Tyr
Trp Thr Asp Glu Trp Ser Val Ala Cys Phe Asp 340
345 350Ser Tyr Asn Ala Ser Ser Pro Ile Phe Thr Asp Thr
Ser Val Gly Asn 355 360 365Pro Val
Asp Arg Gln Trp Glu Trp Phe Leu Cys Asn Glu Pro Phe Phe 370
375 380Trp Trp Gln Asp Gly Ala Pro Glu Gly Thr Ser
Thr Ile Val Pro Arg385 390 395
400Leu Val Ser Ala Ser Tyr Trp Gln Arg Gln Cys Pro Leu Tyr Phe Pro
405 410 415Glu Val Asn Gly
Tyr Thr Tyr Gly Ser Ala Lys Gly Lys Asn Ser Ala 420
425 430Thr Val Asn Ser Trp Thr Gly Gly Trp Asp Met
Thr Arg Asn Thr Thr 435 440 445Arg
Leu Ile Trp Thr Asn Gly Gln Tyr Asp Pro Trp Arg Asp Ser Gly 450
455 460Val Ser Ser Thr Phe Arg Pro Gly Gly Pro
Leu Val Ser Thr Ala Asn465 470 475
480Glu Pro Val Gln Ile Ile Pro Gly Gly Phe His Cys Ser Asp Leu
Tyr 485 490 495Met Glu Asp
Tyr Tyr Ala Asn Glu Gly Val Arg Lys Val Val Asp Asn 500
505 510Glu Val Lys Gln Ile Lys Glu Trp Val Glu
Glu Tyr Tyr Ala 515 520
525170424PRTAspergillus niger 170Met Gln Leu Leu Gln Ser Leu Ile Val Ala
Val Cys Phe Ser Tyr Gly1 5 10
15Val Leu Ser Leu Pro His Gly Pro Ser Asn Gln His Lys Ala Arg Ser
20 25 30Phe Lys Val Glu Arg Val
Arg Arg Gly Thr Gly Ala Leu His Gly Pro 35 40
45Ala Ala Leu Arg Lys Ala Tyr Arg Lys Tyr Gly Ile Ala Pro
Ser Ser 50 55 60Phe Asn Ile Asp Leu
Ala Asp Phe Lys Pro Ile Thr Thr Thr His Ala65 70
75 80Ala Ala Gly Ser Glu Ile Ala Glu Pro Asp
Gln Thr Gly Ala Val Ser 85 90
95Ala Thr Ser Val Glu Asn Asp Ala Glu Phe Val Ser Pro Val Leu Ile
100 105 110Gly Gly Gln Lys Ile
Val Met Thr Phe Asp Thr Gly Ser Ser Asp Phe 115
120 125Trp Val Phe Asp Thr Asn Leu Asn Glu Thr Leu Thr
Gly His Thr Glu 130 135 140Tyr Asn Pro
Ser Asn Ser Ser Thr Phe Lys Lys Met Asp Gly Tyr Thr145
150 155 160Phe Asp Val Ser Tyr Gly Asp
Asp Ser Tyr Ala Ser Gly Pro Val Gly 165
170 175Thr Asp Thr Val Asn Ile Gly Gly Ala Ile Val Lys
Glu Gln Ala Phe 180 185 190Gly
Val Pro Asp Gln Val Ser Gln Ser Phe Ile Glu Asp Thr Asn Ser 195
200 205Asn Gly Leu Val Gly Leu Gly Phe Ser
Ser Ile Asn Thr Ile Lys Pro 210 215
220Glu Ala Gln Asp Thr Phe Phe Ala Asn Val Ala Pro Ser Leu Asp Glu225
230 235 240Pro Val Met Thr
Ala Ser Leu Lys Ala Asp Gly Val Gly Glu Tyr Glu 245
250 255Phe Gly Thr Ile Asp Lys Asp Lys Tyr Gln
Gly Asn Ile Ala Asn Ile 260 265
270Ser Val Asp Ser Ser Asn Gly Tyr Trp Gln Phe Ser Thr Pro Lys Tyr
275 280 285Ser Val Ala Asp Gly Glu Leu
Lys Asp Ile Gly Ser Leu Asn Thr Ser 290 295
300Ile Ala Asp Thr Gly Thr Ser Leu Met Leu Leu Asp Glu Asp Val
Val305 310 315 320Thr Ala
Tyr Tyr Ala Gln Val Pro Asn Ser Val Tyr Val Ser Ser Ala
325 330 335Gly Gly Tyr Ile Tyr Pro Cys
Asn Thr Thr Leu Pro Ser Phe Ser Leu 340 345
350Val Leu Gly Glu Ser Ser Leu Ala Thr Ile Pro Gly Asn Leu
Ile Asn 355 360 365Phe Ser Lys Val
Gly Thr Asn Thr Thr Thr Gly Gln Ala Leu Cys Phe 370
375 380Gly Gly Ile Gln Ser Asn Gly Asn Thr Ser Leu Gln
Ile Leu Gly Asp385 390 395
400Ile Phe Leu Lys Ala Phe Phe Val Val Phe Asp Met Arg Gly Pro Ser
405 410 415Leu Gly Val Ala Ser
Pro Lys Asn 420171548PRTAspergillus niger 171Met Arg Ile Asp
Ser Ala Ala Leu His Leu Val Pro Val Leu Leu Gly1 5
10 15Gln Val Gly Ala Leu Gln Leu Pro Leu Val
Gln Asp Ser Asn Ser Gln 20 25
30Trp Gln Lys Pro Asn Ala Gly Asp Lys Pro Leu Ile Ser Ser Pro Leu
35 40 45Leu Gln Glu Gln Val Lys Ala Glu
Asn Leu Leu Asp Arg Ala Arg Gln 50 55
60Leu Tyr Lys Ile Ala Glu Leu Gly Glu Asp Glu Tyr Asn His Pro Thr65
70 75 80Arg Val Ile Gly Ser
Lys Gly His Leu Gly Thr Leu Asp Tyr Ile Tyr 85
90 95Ser Thr Leu Thr Asp Leu Gly Asp Tyr Tyr Thr
Val Val Asn Gln Ser 100 105
110Phe Pro Ala Val Ser Gly Asn Val Phe Glu Ser Arg Leu Val Leu Gly
115 120 125His Asp Val Pro Lys Ser Ala
Thr Pro Met Gly Leu Thr Pro Pro Thr 130 135
140Arg Asn Lys Glu Pro Val Tyr Gly Ser Leu Val Ala Val Ser Asn
Leu145 150 155 160Gly Cys
Glu Ala Ser Asp Tyr Ser Ser Asn Leu Lys Gly Ala Val Ala
165 170 175Phe Ile Ser Arg Gly Ser Cys
Pro Phe Gly Thr Lys Ser Gln Leu Ala 180 185
190Gly Lys Ala Gly Ala Val Ala Ala Val Ile Tyr Asn Asn Glu
Arg Gly 195 200 205Asp Leu Ser Gly
Thr Leu Gly Asn Pro Thr Pro Asp His Val Ala Thr 210
215 220Phe Gly Ile Ser Asp Glu Asp Ala Ala Pro Val Leu
Glu Lys Leu Asn225 230 235
240Lys Gly Glu Lys Val Asp Ala Ile Ala Tyr Val Asp Ala Ile Val Glu
245 250 255Thr Ile His Thr Thr
Asn Ile Ile Ala Gln Thr Thr Asp Gly Asp Pro 260
265 270Asn Asn Cys Val Met Leu Gly Gly His Ser Asp Ser
Val Ala Glu Gly 275 280 285Pro Gly
Ile Asn Asp Asp Gly Ser Gly Thr Leu Thr Leu Leu Glu Leu 290
295 300Ala Thr Leu Leu Thr Gln Phe Arg Val Asn Asn
Cys Val Arg Phe Ala305 310 315
320Trp Trp Ala Ala Glu Glu Glu Gly Leu Leu Gly Ser Asp Tyr Tyr Val
325 330 335Ser Val Leu Thr
Pro Glu Glu Asn Arg Lys Ile Arg Leu Phe Met Asp 340
345 350Tyr Asp Met Leu Gly Ser Pro Asn Phe Ala Tyr
Gln Val Tyr Asn Ala 355 360 365Thr
Asn Ala Val Asn Pro Glu Gly Ser Glu Glu Leu Arg Asp Leu Tyr 370
375 380Thr Asp Phe Tyr Glu Asp His Gly Phe Asn
Tyr Thr Tyr Ile Pro Phe385 390 395
400Asp Gly Arg Ser Asp Tyr Asp Ala Phe Ile Arg His Gly Ile Pro
Gly 405 410 415Gly Gly Ile
Ala Thr Gly Ala Glu Gly Ile Lys Thr Val Glu Glu Ala 420
425 430Asp Met Phe Gly Gly Val Ala Gly Gln Trp
Tyr Asp Pro Cys Tyr His 435 440
445Gln Ile Cys Asp Thr Val Ala Asn Val Asn Leu Thr Ala Trp Glu Trp 450
455 460Asn Thr Lys Leu Val Ala His Ser
Ile Ala Thr Tyr Ala Lys Ser Phe465 470
475 480Asp Gly Phe Pro Glu Arg Ser Asp Glu Pro Ile Ser
Pro Ala Ala Phe 485 490
495Glu Glu Pro Lys Tyr His Gly His Ala Leu Gln Leu Leu Arg Gly Asn
500 505 510Thr Thr Gly Thr Gln Ser
Val Leu Trp Gly Ala Gln Ile Gln Asn Gly 515 520
525Thr Ala Ala Ser Val Leu Asn Leu Leu Ser Ile Arg Arg Arg
Gly Thr 530 535 540Phe Ser Leu Ser545
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