Study on Mutations of Cephalosporin C Acylase

doi:10.13523/j.cb.20150209

China Biotechnology

2015, Vol. 35

Issue (2): 59-65 DOI: 10.13523/j.cb.20150209

Study on Mutations of Cephalosporin C Acylase

XU Xue-li^1,2, ZHANG Wei¹, LIU Yan¹, XIE Li-ping¹, HU You-jia¹

1. China State Institute of Pharmaceutical Industry Zhangjiang Institute, Shanghai 201203, China;
2. Shanghai Institute of Pharmaceutical Industry, Shanghai 200437, China

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Abstract

Cephalosporin C acylase can directly convert cephalosporin C (CPC) to 7-aminocephalosporanic acid (7-ACA), and this one-step enzymatic process has great economic value. A lot of researchers pay attention on it, especially on how to improve the activity and specificity of CPC acylase. The reported CPC acylase mutant S12 has an improved activity that contains 6 point mutations. On the basis of ecs50 gene, six individual mutations (V122A,G140S,F297N,I314T,I415V,S710L) were obtained by using the overlap PCR. Then recombinant proteins were purified after induction, enzymatic activities were tested to study the relationship between different mutation site and the activity of CPC acylase. The result showed that mutant V122A had an increase activity by 23.7% compared to ECS50.

Key words： CPC acylase Overlap extension PCR Enzyme activity Cephalosporin C 7-aminocephalosporanic acid

Received: 11 November 2014 Published: 25 February 2015

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Cite this article:

XU Xue-li, ZHANG Wei, LIU Yan, XIE Li-ping, HU You-jia. Study on Mutations of Cephalosporin C Acylase. China Biotechnology, 2015, 35(2): 59-65.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150209 OR https://manu60.magtech.com.cn/biotech/Y2015/V35/I2/59

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