文献类型：期刊文献

英文题名：Cloning, expression, and characterization of an anti-Prelog stereospecific carbonyl reductase from Gluconobacter oxydans DSM2343

作者：Chen, Rong[1,2];Liu, Xu[1];Wang, Jiale[1];Lin, Jinping[1];Wei, Dongzhi[1]

机构：[1]E China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Hangzhou Normal Univ, Ctr Biomed & Hlth, Div Basical Med, Hangzhou 310012, Zhejiang, Peoples R China

年份：2015

卷号：70

起止页码：18

外文期刊名：ENZYME AND MICROBIAL TECHNOLOGY

收录：;EI(收录号：20150300434660);WOS:【SCI-EXPANDED(收录号:WOS:000350094500003)】；

基金：This work was financially supported by the National Key Basic Research Development Program of China ("973" Program, No. 2012CB721003), the Natural Science Foundation of China (No. 21276084 and 21002029), National Major Science and Technology Projects of China (No. 2012ZX09304009), and Technology Research and Development Program for Institute of Hangzhou (20130432B05).

语种：英文

外文关键词：Carbonyl reductase; Asymmetric reduction; Anti-Prelog rule; Molecular modeling

摘要：A new anti-Prelog stereospecific carbonyl reductase (GoKR) from Gluconobacter oxydans DSM2343 was cloned and identified in Escherichia coli. This GoKR formed a homo-tetramer with a subunit size of approximately 27.0 kDa. GoKR exhibited full activity with NADPH but not with NADH as a cofactor. The optimal pH and temperature were 9.0 and 30 degrees C, respectively. GoKR reduced various ketones, including aliphatic and aromatic ketones, alpha- and beta-keto esters. Aromatic ketones were reduced to (R)-enantiomers, whereas keto esters were reduced to (S)-hydroxy esters with different enantioselectivities. The data indicate that GoKR does not obey Prelog's rule and exhibits anti-Prelog enantiopreference. Enzyme-substrate-cofactor docking analysis showed that hydride transfer occurred at the si faces of carbonyl group for ethyl 4-chloro-3-oxobutanoate (COBE), which was then selectively reduced to the chiral (S)-alcohol. Excellent enantioselectivities were obtained for reducing CUBE and ethyl 2-oxo-4-phenylbutyrate into the corresponding (S)-type products. These products are important for synthesizing HMG-CoA reductase (statins) and angiotensin-converting enzyme inhibitors, respectively. (C) 2014 Elsevier Inc. All rights reserved.