A Novel Thermal Stable Carbonyl Reductase from Bacillus cereus by Gene Mining as Biocatalyst for β-Carbonyl Ester Asymmetric Reduction Reaction

Carbonyl reductase, as biocatalyst, is very important to chiral alcohols production through asymmetric reduction of carbonyl compound. A novel thermal stable carbonyl reductase from Bacillus cereus (BcCR) dependant on NADPH was obtained through a new genome mining strategy proposed in this work. By...

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Bibliographic Details
Published in:Catalysis letters 2019-02, Vol.149 (2), p.610-618
Main Authors: Qin, Yan-Li, Ruan, Tao, Hou, He-Shuai, Hou, Ya-Li, Yang, Zhong-Hua, Quan, Can
Format: Article
Language:English
Subjects:
Alcohols
Asymmetry
Biocatalysts
Carbonyl compounds
Carbonyls
Catalysis
Chemical reduction
Chemistry
Chemistry and Materials Science
Deoxyribonucleic acid
DNA
E coli
Gene expression
Industrial Chemistry/Chemical Engineering
Organometallic Chemistry
Physical Chemistry
Reductases
Thermal stability
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Summary:Carbonyl reductase, as biocatalyst, is very important to chiral alcohols production through asymmetric reduction of carbonyl compound. A novel thermal stable carbonyl reductase from Bacillus cereus (BcCR) dependant on NADPH was obtained through a new genome mining strategy proposed in this work. By analyzing its amino acid sequence and structure, the BcCR should be a thermal stability and wide pH tolerance carbonyl reductase. Its gene was cloned by PCR with B. cereus genomic DNA as template. Its heterologous expression system, E. coli BL21 (DE3) plysS/pET28a- bccr , was constructed, and BcCR was successfully expressed. Enzymatic properties show that at 57.5 °C and pH 7.0 it can reach maximum reaction rate. Its K m and V max to ethyl 4-chloroacetoacetate is 1.85 mmol/L and 0.22 µmol/(min·mg protein ), respectively. It can catalyze the asymmetric reduction of the β-carbonyl compound, such as ethyl 4-chloroacetoacetate to ethyl S -4-chloro-3-hydroxybutyrate. This paper proposes a practical method for discovery of new carbonyl reductases, and provides a novel enzyme as biocatalyst for asymmetric reduction of β-carbonyl compound.
ISSN:1011-372X
1572-879X
DOI:10.1007/s10562-018-2645-4