Journal
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
Volume 78, Issue 8, Pages 1350-1356Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/09168451.2014.925775
Keywords
carbonyl reductase; enantioselective reduction; chiral alcohols
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Funding
- National Key Basic Research Development Program of China [973 Program] [2012CB721003]
- Natural Science Foundation of China [21002029/B020706, 21276084/B060804]
- National Major Science and Technology Projects of China [2012ZX09304009]
- Natural Science Foundation of Zhejiang Province [LY13C050004]
- Science and Technology Development Foundation of Hangzhou [20101131N05]
- Technology Research and Development Program for Institute of Hangzhou [20130432B05]
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The versatile carbonyl reductases from Gluconobacter oxydans in the enantioselective reduction of ketones to the corresponding alcohols were exploited by genome search approach. All purified enzymes showed activities toward the tested ketoesters with different activities. In the reduction of 4-phenyl-2-butanone with in situ NAD(P)H regeneration system, (S)-alcohol was obtained with an e.e. of up to 100% catalyzed by Gox0644. Under the same experimental condition, all enzymes catalyzed ethyl 4-chloroacetoacetate to give chiral products with an excellent e.e. of up to 99%, except Gox0644. Gox2036 had a strict requirement for NADH as the cofactor and showed excellent enantiospecificity in the synthesis of ethyl (R)-4-chloro-3-hydroxybutanoate. For the reduction of ethyl 2-oxo-4-phenylbutyrate, excellent e.e. (>99%) and high conversion (93.1%) were obtained by Gox0525, whereas the other enzymes showed relatively lower e.e. and conversions. Among them, Gox2036 and Gox0525 showed potentials in the synthesis of chiral alcohols as useful biocatalysts.
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