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

Dengchao Li, Yewang Zhang, Shiwei Cheng, Qiong Gao and 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

Article history:

Received March 30, 2006

Accepted March 26, 2007

Key words:

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


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.


*Corresponding author: 
                                               ++86 21 642 52 981
                                               ++86 21 642 50 068

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