文献类型：期刊文献

英文题名：Highly stereoselective reduction of prochiral ketones by a bacterial reductase coupled with cofactor regeneration

作者：Ni, Yan[1];Li, Chun-Xiu[1];Wang, Li-Juan[1];Zhang, Jie[1];Xu, Jian-He[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Biocatalysis & Synthet Biotechnol, Shanghai 200237, Peoples R China

年份：2011

卷号：9

期号：15

起止页码：5463

外文期刊名：ORGANIC & BIOMOLECULAR CHEMISTRY

收录：;EI(收录号：20112914157772);WOS:【SCI-EXPANDED(收录号:WOS:000292983400022)】；

基金：This work was financially supported by the National Natural Science Foundation of China (Nos. 20902023 & 31071604), Ministry of Science and Technology, P. R. China (Nos. 2011CB710800 & 2009ZX09501-016), and China National Special Fund for State Key Laboratory of Bioreactor Engineering (No. 2060204).

语种：英文

外文关键词：Aromatic compounds - Bacteriology - Enantiomers - Ketones

摘要：A carbonyl reductase gene (yueD) from Bacillus sp. ECU0013 was heterologously overexpressed in Escherichia coli, and the encoded protein (BYueD) was purified to homogeneity and characterized. The NADPH-dependent reductase showed a broad substrate spectrum towards different aromatic ketones, and alpha-and beta-ketoesters. Although the enantioselectivity was high to moderate for the reduction of alpha-ketoesters, all the tested beta-ketoesters and aromatic ketones were reduced to the corresponding chiral alcohols in enantiomerically pure forms. Furthermore, the practical applicability of this enzyme was evaluated for the reduction of ethyl 4-chloro-3-oxobutanoate (1a). Using Escherichia coli cells coexpressing BYueD and glucose dehydrogenase, 215 g L-1 (1.3 M) of 1a was stoichiometrically converted to ethyl (R)-4-chloro-3-hydroxybutanoate ((R)-1b) in an aqueous-toluene biphasic system by using a substrate fed-batch strategy, resulting in an overall hydroxyl product yield of 91.7% with enantiomeric purity of 99.6% ee.