文献类型：期刊文献

英文题名：Efficient production of ethyl (R)-4-chloro-3-hydroxybutanoate by a novel alcohol dehydrogenase from Lactobacillus curieae S1L19

作者：Zhang, Yiping[1];Wang, Hualei[1];Chen, Lifeng[1];Wu, Kai[1];Xie, Jingli[1];Wei, Dongzhi[1]

机构：[1]East China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2016

卷号：134

起止页码：51

外文期刊名：JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC

收录：;EI(收录号：20163902845842);WOS:【SCI-EXPANDED(收录号:WOS:000391074600008)】；

基金：This work was supported by the National Natural Science Foundation of China (Grant No. 21406068/B060804), the Fundamental Research Funds for the Central Universities, and the National Basic Research Program of China (Grant No. 2012CB721103).

语种：英文

外文关键词：Cofactor; Co-expression; Asymmetric reduction; Ethyl-(R)-4-chloro-3-hydroxybutanoate

摘要：Ethyl (R)-4-chloro-3-hydroxybutanoate ester [(R)-CHBE] is an important chiral intermediate for the synthesis of chiral drugs. In this study, a novel short -chain, NADH-dependent dehydrogenase (LCRIII) from Lactobacillus curieae S1L19 was discovered to exhibit high activity and enantioselectivity in the production of (R)-CHBE by reduction of ethyl 4-chloroacetoacetate (COBE). LCRIII was heterologously overexpressed in Escherichia coli and the protein was purified to homogeneity. Characterization of LCRIII showed broad substrate specificity towards a variety of ketones. In addition, an efficient cofactor regeneration system was constructed by co-expressing LCRIII and glucose dehydrogenase (GDH) in E. coli cells. Up to 1.5 M (246.8 g/L) COBE could be completely reduced to (R)-CHBE with excellent enantiomeric excess ( > 99% ee) in a monophasic aqueous system. Moreover, the process could be performed even without external addition of cofactors. These results demonstrate the great potential of this process in industrial applications. (C) 2016 Elsevier B.V. All rights reserved.