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Process optimization for enzymatic synthesis of cis-cefprozil by a mutant penicillin G acylase*(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2016 03
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Process optimization for enzymatic synthesis of cis-cefprozil by a mutant penicillin G acylase*
ZHAO Yan12 ZHANG Dalong2 LI Jianjiong2 FENG Jinhui2 BAO Jinku1** WU Qiaqing2** & ZHU Dunming2
1College of Life Sciences, Sichuan University, Chengdu 610064, China 2Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China
penicillin G acylase mutants cis-cefprozil immobilization
TQ464 : Q939.9

Cefprozil is a second generation cephalosporin antibiotics widely used in the treatment of upper and lower respiratory tract infections as well as skin and soft tissue infections caused by sensitive bacteria. Because of the great advantages of biocatalysis over the conventional chemical synthesis by avoiding toxic reagents, solvents and harsh reaction conditions, it is desirable to develop a feasible enzymatic synthesis of cefprozil. In this work, we chose the key sites βF24、αF146 of pecicillin G acylase from Escherichia coli for site-directed mutagenesis. By comparing the synthesis efficiency of WT, βF24A and αF146Y/βF24A, the obtained mutant βF24A which has high synthesis/hydrolysis ratio was used in the enzymatic synthesis of cis-cefprozil from cis-7-APRA and d-HPGME. Based on the aqueous-phase synthesis system, the yield was dependent on the multiple factors. The most suitable temperature and pH was 25 °C and 6.0, respectively, the optimum molar ratio of d-HPGME/cis-7-APRA 3:1 and the enzyme loading 2.6 U/mL. At the optimized conditions, 99% yield was attained with 100 mmol/L of cis-7-APRA. The maximum yield was 99% with immobilized βF24A at the optimized conditions with 100 mmol/L cis-7-APRA. Yields remained 52% after 60 consecutive batch reactions. This paper studies on the selection of mutants and optimization for enzyme catalysis conditions, which provides useful information for enzymatic synthesis of cis-cefprozil.


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Last Update: 2016-06-25