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Catalysis Letters

, Volume 149, Issue 2, pp 610–618 | Cite as

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

  • Yan-Li Qin
  • Tao Ruan
  • He-Shuai Hou
  • Ya-Li Hou
  • Zhong-Hua YangEmail author
  • Can QuanEmail author
Article
  • 123 Downloads

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.

Keywords

Gene mining Carbonyl reductase Bacillus cereus Asymmetric reduction Chiral alcohol 

Notes

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).

Compliance with Ethical Standards

Conflict of interest Statement

The authors declare no conflict of interest.

Supplementary material

10562_2018_2645_MOESM1_ESM.docx (3.1 mb)
Supplementary material 1 (DOCX 3147 KB)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Chemistry and Chemical EngineeringWuhan University of Science and TechnologyWuhanChina
  2. 2.Division of Chemical Metrology and Analytical ScienceNational Institute of MetrologyBeijingChina
  3. 3.Key Laboratory of Hubei Province for Coal Conversion and New Carbon MaterialsWuhan University of Science and TechnologyWuhanChina

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