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Engineering of a novel carbonyl reductase with coenzyme regeneration in E. coli for efficient biosynthesis of enantiopure chiral alcohols.
J Biotechnol. 2016 Jul 20; 230:54-62.JB

Abstract

The novel anti-Prelog stereospecific carbonyl reductase from Acetobacter sp. CCTCC M209061 was successfully expressed in E. coli combined with glucose dehydrogenase (GDH) to construct an efficient whole-cell biocatalyst with coenzyme NADH regeneration. The enzymatic activity of GAcCR (AcCR with a GST tag) reached 304.9U/g-dcw, even 9 folds higher than that of wild strain, and the activity of GDH for NADH regeneration recorded 46.0U/mg-protein in the recombinant E. coli. As a whole-cell biocatalyst, the recombinant E. coli BL21(DE3)pLysS (pETDuet-gaccr-gdh) possessed a broad substrate spectrum for kinds of carbonyl compounds with encouraging yield and stereoselectivity. Besides, the asymmetric reduction of ethyl 4-chloroacetoacetate (COBE) to optically pure ethyl 4-chloro-3-hydroxybutyrate (CHBE) catalyzed by the whole-cell biocatalyst was systematically investigated. Under the optimal reaction conditions, the optical purity of CHBE was over 99% e.e. for (S)-enantiomer, and the initial rate and product yield reached 8.04μmol/min and 99.4%, respectively. Moreover, the space-time yield was almost 20 folds higher than that catalyzed by the wild strain. Therefore, a new, high efficiency biocatalyst for asymmetric reductions was constructed successfully, and the enantioselective reduction of prochiral compounds using the biocatalyst was a promising approach for obtaining enantiopure chiral alcohols.

Authors+Show Affiliations

Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, Guangdong, China; School of Chemistry and Chemical Engineering, South China University of Technology, Guangzhou 510640, Guangdong, China.Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, Guangdong, China.State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, Guangdong, China.Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, Guangdong, China; School of Chemistry and Chemical Engineering, South China University of Technology, Guangzhou 510640, Guangdong, China.Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, Guangdong, China. Electronic address: wylou@scut.edu.cn.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27211999

Citation

Wei, Ping, et al. "Engineering of a Novel Carbonyl Reductase With Coenzyme Regeneration in E. Coli for Efficient Biosynthesis of Enantiopure Chiral Alcohols." Journal of Biotechnology, vol. 230, 2016, pp. 54-62.
Wei P, Gao JX, Zheng GW, et al. Engineering of a novel carbonyl reductase with coenzyme regeneration in E. coli for efficient biosynthesis of enantiopure chiral alcohols. J Biotechnol. 2016;230:54-62.
Wei, P., Gao, J. X., Zheng, G. W., Wu, H., Zong, M. H., & Lou, W. Y. (2016). Engineering of a novel carbonyl reductase with coenzyme regeneration in E. coli for efficient biosynthesis of enantiopure chiral alcohols. Journal of Biotechnology, 230, 54-62. https://doi.org/10.1016/j.jbiotec.2016.05.004
Wei P, et al. Engineering of a Novel Carbonyl Reductase With Coenzyme Regeneration in E. Coli for Efficient Biosynthesis of Enantiopure Chiral Alcohols. J Biotechnol. 2016 Jul 20;230:54-62. PubMed PMID: 27211999.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Engineering of a novel carbonyl reductase with coenzyme regeneration in E. coli for efficient biosynthesis of enantiopure chiral alcohols. AU - Wei,Ping, AU - Gao,Jia-Xin, AU - Zheng,Gao-Wei, AU - Wu,Hong, AU - Zong,Min-Hua, AU - Lou,Wen-Yong, Y1 - 2016/05/17/ PY - 2016/02/09/received PY - 2016/04/24/revised PY - 2016/05/03/accepted PY - 2016/5/24/entrez PY - 2016/5/24/pubmed PY - 2017/2/25/medline KW - (S)-CHBE KW - Acetobacter sp KW - Asymmetric reduction KW - Chiral alcohols KW - GAcCR KW - Whole-cell biocatalysis SP - 54 EP - 62 JF - Journal of biotechnology JO - J. Biotechnol. VL - 230 N2 - The novel anti-Prelog stereospecific carbonyl reductase from Acetobacter sp. CCTCC M209061 was successfully expressed in E. coli combined with glucose dehydrogenase (GDH) to construct an efficient whole-cell biocatalyst with coenzyme NADH regeneration. The enzymatic activity of GAcCR (AcCR with a GST tag) reached 304.9U/g-dcw, even 9 folds higher than that of wild strain, and the activity of GDH for NADH regeneration recorded 46.0U/mg-protein in the recombinant E. coli. As a whole-cell biocatalyst, the recombinant E. coli BL21(DE3)pLysS (pETDuet-gaccr-gdh) possessed a broad substrate spectrum for kinds of carbonyl compounds with encouraging yield and stereoselectivity. Besides, the asymmetric reduction of ethyl 4-chloroacetoacetate (COBE) to optically pure ethyl 4-chloro-3-hydroxybutyrate (CHBE) catalyzed by the whole-cell biocatalyst was systematically investigated. Under the optimal reaction conditions, the optical purity of CHBE was over 99% e.e. for (S)-enantiomer, and the initial rate and product yield reached 8.04μmol/min and 99.4%, respectively. Moreover, the space-time yield was almost 20 folds higher than that catalyzed by the wild strain. Therefore, a new, high efficiency biocatalyst for asymmetric reductions was constructed successfully, and the enantioselective reduction of prochiral compounds using the biocatalyst was a promising approach for obtaining enantiopure chiral alcohols. SN - 1873-4863 UR - https://www.unboundmedicine.com/medline/citation/27211999/Engineering_of_a_novel_carbonyl_reductase_with_coenzyme_regeneration_in_E__coli_for_efficient_biosynthesis_of_enantiopure_chiral_alcohols_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0168-1656(16)30259-0 DB - PRIME DP - Unbound Medicine ER -