Discovery and protein engineering of penicillin G acylase for biosynthesis of cefradine.

Lingyi LIU; Xiangying LI; Congcong LI; Lijuan MA; Bo YUAN; Zhoutong SUN

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Discovery and protein engineering of penicillin G acylase for biosynthesis of cefradine.

Author: Lingyi LIU ¹ ; Xiangying LI ² ; Congcong LI ² ; Lijuan MA ¹ ; Bo YUAN ² ; Zhoutong SUN ²
Author Information

1. School of Biological Engineering, Tianjin University of Science and Technology, Tianjin 300457, China.
2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
Publication Type:Journal Article
Keywords: cefradine; continuous flow reactions; enzymatic catalysis; enzyme immobilization; penicillin G acylase
MeSH: Penicillin Amidase/biosynthesis*; Protein Engineering/methods*; Cephradine/metabolism*; Escherichia coli/metabolism*; Enzymes, Immobilized/metabolism*; Recombinant Proteins/biosynthesis*
From: Chinese Journal of Biotechnology 2025;41(9):3630-3642
CountryChina
Language:Chinese
Abstract: Penicillin G acylases (PGAs) are industrially important enzymes primarily used for the synthesis of first- and second-generation cephalosporins or penicillins. This study aims to establish a high-efficiency biosynthetic system for cefradine on the purpose of significantly enhancing its catalytic efficiency in cefradine synthesis and developing its potentials for industrial application. In this study, we identified and engineered penicillin G acylase and obtained a highly active mutant KsPGA M7(M168F/F313G) for the synthesis of cefradine. The mutant achieved a conversion rate over 95% in the scaled-up reaction. To validate its industrial applicability, we immobilized both the wild-type and mutant enzymes and applied them in continuous flow reactions, which achieved a space-time yield of 2 800 g/(L·d). This study lays a foundation for the future applications of penicillin G acylases in the industrial synthesis of cefradine.