title: Iterative combinatorial mutagenesis as an effective strategy for generation of deacetoxycephalosporin C synthase with improved activity toward penicillin G.
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.
authors.affiliation: School of Chemical and Environmental Engineering, China University of Mining and Technology, Beijing, People's Republic of China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Ji
authors.forename: Keqiang
authors.initials: K
authors.lastname: Fan
authors.forename: Xiuyun
authors.initials: X
authors.lastname: Tian
authors.forename: Xia
authors.initials: X
authors.lastname: Zhang
authors.forename: Yuxiu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Keqian
authors.initials: K
authors.lastname: Yang
mesh_terms: D055162:Biocatalysis; D019751:Intramolecular Transferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D007700:Kinetics; D016296:Mutagenesis; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010400:Penicillin G / Q000378:metabolism*; D046915:Penicillin-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010406:Penicillins / Q000378:metabolism*; D015202:Protein Engineering; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics / Q000378:metabolism
chemical_list: D046915:Penicillin-Binding Proteins; D010406:Penicillins; D019751:Intramolecular Transferases; C038371:deacetoxycephalosporin C synthetase; D010400:Penicillin G