Directed evolution of Streptomyces clavuligerus deacetoxycephalosporin C synthase for enhancement of penicillin G expansion

Article Abstract:

The deacetoxycephalosporin C synthase from Streptomyces clavuligerus is directly modified for enhancement of penicillin G expansion into phenylacetyl-7-aminodeacetoxycephalosporanic acid. The analysis has provided promising enzymes for cephalosporin biosynthesis along with a large number of mutants, which has provided new insights into the structure-function relationship of the protein that should lead to further rational engineering.

User Contributions:

Comment about this article or add new information about this topic:

Comment:  (50-4000 characters)

Name:

E-mail:

Security Code:

Family shuffling of expandase genes to enhance substrate specificity for penicillin G

Article Abstract:

Screening of several new expandase genes from actinomycetes and enhancing the substrate specificity for penicillin G using DNA family shuffling is reported. The evolution of penicillin G-specific expandase was successfully directed and the results indicate the remarkable plasticity of the expandase, suggesting that more-active chimeras might be achievable with further rounds.

User Contributions:

Comment about this article or add new information about this topic:

Comment:  (50-4000 characters)

Name:

E-mail:

Security Code:

Engineering streptomyces clavuligerus deacetoxycephalosporin C synthase for optimal ring expansion activity toward penicillin G

Article Abstract:

Three mutants each by random- and site-directed mutagenesis of the Streptomyces clavuligerus show two- to sixfold and 6- to 14-fold increase in the catalysis rate of deacetoxycephalosporin C enzyme activity than that of wild-type, respectively. Furthermore, double and triple mutants show even higher enhancement of enzyme activity and reactivity toward pencillin G.

User Contributions:

Comment about this article or add new information about this topic:

Comment:  (50-4000 characters)

Name:

E-mail:

Security Code: