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Updated: May 19, 2026

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
Published on: March 16, 2011
Iterative combinatorial mutagenesis as an effective strategy for generation of deacetoxycephalosporin C synthase with
Junjie Ji1, Keqiang Fan, Xiuyun Tian
1School of Chemical and Environmental Engineering, China University of Mining and Technology, Beijing, People's Republic of China.
Abstract:
An iterative combinatorial mutagenesis (ICM) strategy was used to engineer deacetoxycephalosporin C synthase of Streptomyces clavuligerus (scDAOCS) for improved activity toward penicillin G. Seven mutational sites were repeatedly combined onto a starter mutant (C155Y Y184H V275I C281Y) of scDAOCS. Eleven improved combinatorial mutants were identified from 24 mutants in four rounds of ICM.
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