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Enhanced Enzymatic Production of Cephalexin at High Substrate Concentration with in situ Product Removal by Complexation

Dengchao Li ; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China, University of Science and Technology, CN-200237 Shanghai, PR China
Yewang Zhang ; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China, University of Science and Technology, CN-200237 Shanghai, PR China
Shiwei Cheng ; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China, University of Science and Technology, CN-200237 Shanghai, PR China
Qiong Gao ; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China, University of Science and Technology, CN-200237 Shanghai, PR China
Dongzhi Wei ; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China, University of Science and Technology, CN-200237 Shanghai, PR China


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Abstract

Cephalexin (CEX) was synthesized with 7-amino-3-deacetoxycephalosporanic acid (7-ADCA) and D(–)-phenylglycine methyl ester (PGME) using immobilized penicillin G acylase from Escherichia coli. It was found that substrate concentration and in situ product could remarkably influence the ratio of synthesis to hydrolysis (S/H) and the efficiency of CEX synthesis. The optimal ratio of enzyme to substrate was 65 IU/mM 7-ADCA. High substrate concentration improved the 7-ADCA conversion from 61 to 81 % in the process without in situ product removal (ISPR), while in the synthetic process with ISPR, high substrate concentration increased the 7-ADCA conversion from 88 to 98 %. CEX was easily separated from CEX/β-naphthol complex and its purity and overall yield were 99 and 70 %, respectively.

Keywords

cephalexin; complexation; enzymatic synthesis; in situ product removal; penicillin G acylase

Hrčak ID:

30425

URI

https://hrcak.srce.hr/30425

Publication date:

17.11.2008.

Article data in other languages: croatian

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