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

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Abstract

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 Km and Vmax to ethyl 4-chloroacetoacetate is 1.85 mmol/L and 0.22 µmol/(min·mgprotein), 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.

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Acknowledgements

The authors acknowledge all the financial supports for this research by the National Natural Science Foundation of China (Grant No. 21376184), the Scientific Research Foundation for the Returned Overseas Chinese Scholars (State Education Ministry), Foundation from the Educational Commission of Hubei Province of China (Grant No. D20121108), the Ministry of Science and Technology of China (2017YFF0205803), and the National Institute of Metrology of China (21-AKY1615).

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Author notes

  1. Yan-Li Qin and Tao Ruan contributed equally to this work.

  2. Yan-Li Qin and Tao Ruan should be considered as co-first authors.

Authors and Affiliations

  1. School of Chemistry and Chemical Engineering, Wuhan University of Science and Technology, Wuhan, 430081, China

    Yan-Li Qin, Tao Ruan, He-Shuai Hou, Ya-Li Hou & Zhong-Hua Yang

  2. Division of Chemical Metrology and Analytical Science, National Institute of Metrology, Beijing, 100029, China

    Can Quan

  3. Key Laboratory of Hubei Province for Coal Conversion and New Carbon Materials, Wuhan University of Science and Technology, Wuhan, 430081, China

    Zhong-Hua Yang

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  1. Yan-Li Qin
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  2. Tao Ruan
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  3. He-Shuai Hou
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  4. Ya-Li Hou
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Qin, YL., Ruan, T., Hou, HS. et al. A Novel Thermal Stable Carbonyl Reductase from Bacillus cereus by Gene Mining as Biocatalyst for β-Carbonyl Ester Asymmetric Reduction Reaction. Catal Lett 149, 610–618 (2019). https://doi.org/10.1007/s10562-018-2645-4

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