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头孢菌素 C 酰化酶的生物合成:最佳培养基设计、纯化及特性分析

Biosynthesis of cephalosporin-C acylase enzyme: optimal media design, purification, and characterization.

作者信息

Gaurav Kumar, Kundu Kanika, Kundu Subir

机构信息

School of Biochemical Engineering, Institute of Technology, Banaras Hindu University, Varanasi, India.

出版信息

Artif Cells Blood Substit Immobil Biotechnol. 2010 Oct;38(5):277-83. doi: 10.3109/10731199.2010.482036.

Abstract

7-aminocephalosporanic acid (7-ACA) is the key intermediate for the synthesis of semisynthetic cephalosporin antibiotics and enzyme cephalosporin-C acylase (CPC acylase) plays an important role in the conversion of cephalosporin-C to 7-ACA. With an aim to increase the yield of 7-ACA production by Micrococcus luteus, a stepwise strategy, statistical medium was applied for optimizing the medium composition for the production of CPC acylase. Purified enzyme was found to be of 80 kDa. The optimum pH and temperature for the production of 7-ACA were 7.6 and 340C, respectively. The Km and Vmax were estimated to be 9.43 mg/mL and 7.65 U/mL, respectively.

摘要

7-氨基头孢烷酸(7-ACA)是合成半合成头孢菌素抗生素的关键中间体,而酶头孢菌素-C酰基转移酶(CPC酰基转移酶)在头孢菌素-C转化为7-ACA的过程中起着重要作用。为了提高藤黄微球菌生产7-ACA的产量,采用逐步策略,利用统计培养基优化用于生产CPC酰基转移酶的培养基组成。发现纯化后的酶分子量为80 kDa。生产7-ACA的最适pH值和温度分别为7.6和34℃。米氏常数(Km)和最大反应速度(Vmax)估计分别为9.43 mg/mL和7.65 U/mL。

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