文献类型：期刊文献

英文题名：Engineering of a novel carbonyl reductase with coenzyme regeneration in E. coli for efficient biosynthesis of enantiopure chiral alcohols

作者：Wei, Ping[1];Gao, Jia-Xin[1];Zheng, Gao-Wei[2];Wu, Hong[1];Zong, Min-Hua[1,3];Lou, Wen-Yong[1]

机构：[1]S China Univ Technol, Sch Food Sci & Engn, Lab Appl Biocatalysis, Guangzhou 510640, Guangdong, Peoples R China;[2]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[3]S China Univ Technol, Sch Chem & Chem Engn, Guangzhou 510640, Guangdong, Peoples R China

年份：2016

卷号：230

起止页码：54

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20162202444556);WOS:【SCI-EXPANDED(收录号:WOS:000377981300012)】；

基金：We wish to thank the National Natural Science Foundation of China (21336002; 21376096), the Key Program of Guangdong Natural Science Foundation (S2013020013049), the Fundamental Research Funds for the Chinese Universities (2015PT002; 2015ZP009), and the Open Funding Project of the State Key Laboratory of Bioreactor Engineering for partially funding this work.

语种：英文

外文关键词：Asymmetric reduction; Whole-cell biocatalysis; Acetobacter sp; GAcCR; (S)-CHBE; Chiral alcohols

摘要：The novel anti-Prelog stereospecific carbonyl reductase from Acetobacter sp. CCTCC M209061 was successfully expressed in E. coli combined with glucose dehydrogenase (GDH) to construct an efficient whole-cell biocatalyst with coenzyme NADH regeneration. The enzymatic activity of GAcCR (AcCR with a GST tag) reached 304.9 U/g-dcw, even 9 folds higher than that of wild strain, and the activity of GDH for NADH regeneration recorded 46.0 U/mg-protein in the recombinant E. coli. As a whole-cell biocatalyst, the recombinant E. coli BL21(DE3)pLysS (pETDuet-gaccr-gdh) possessed a broad substrate spectrum for kinds of carbonyl compounds with encouraging yield and stereoselectivity. Besides, the asymmetric reduction of ethyl 4-chloroacetoacetate (COBE) to optically pure ethyl 4-chloro-3-hydroxybutyrate (CHBE) catalyzed by the whole-cell biocatalyst was systematically investigated. Under the optimal reaction conditions, the optical purity of CHBE was over 99% e.e. for (S)-enantiomer, and the initial rate and product yield reached 8.04 mu mol/min and 99.4%, respectively. Moreover, the space-time yield was almost 20 folds higher than that catalyzed by the wild strain. Therefore, a new, high efficiency biocatalyst for asymmetric reductions was constructed successfully, and the enantioselective reduction of prochiral compounds using the biocatalyst was a promising approach for obtaining enantiopure chiral alcohols. (C) 2016 Elsevier B.V. All rights reserved.