期刊
ENZYME AND MICROBIAL TECHNOLOGY
卷 70, 期 -, 页码 18-27出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.enzmictec.2014.12.004
关键词
Carbonyl reductase; Asymmetric reduction; Anti-Prelog rule; Molecular modeling
资金
- National Key Basic Research Development Program of China (973 Program) [2012CB721003]
- Natural Science Foundation of China [21276084, 21002029]
- National Major Science and Technology Projects of China [2012ZX09304009]
- Technology Research and Development Program for Institute of Hangzhou [20130432B05]
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.
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