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Effective Biotransformation of Ethyl 4-Chloro-3-Oxobutanoate into Ethyl (S)-4-Chloro-3-Hydroxybutanoate by Recombinant E. coli CCZU-T15 Whole Cells in [ChCl][Gly]–Water Media

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Abstract

To increase the biocatalytic activity of Escherichia coli CCZU-T15 whole cells, choline chloride/glycerol ([ChCl][Gly]) was firstly used as biocompatible solvent for the effective biotransformation of ethyl 4-chloro-3-oxobutanoate (COBE) into ethyl (S)-4-chloro-3-hydroxybutanoate [(S)-CHBE]. Furthermore, L-glutamine (150 mM) was added into [ChCl][Gly]–water ([ChCl][Gly] 12.5 vol%, pH 6.5) media instead of NAD+ for increasing the biocatalytic efficiency. To further improve the biosynthesis of (S)-CHBE (>99 % e.e.) by E. coli CCZU-T15 whole cells, Tween-80 (7.5 mM) was also added into this reaction media, and (S)-CHBE (>9 % e.e.) could be effectively synthesized from 2000 and 3000 mM COBE in the yields of 100 and 93.0 % by whole cells of recombinant E. coli CCZU-T15, respectively. TEM image indicated that the cell membrane was permeabilized and lost its integrity and when the cell was exposed to [ChCl][Gly]–water media with Tween-80. Clearly, this bioprocess has high potential for the effective biosynthesis of (S)-CHBE (>99 % e.e.).

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (No. 21102011), Higher Education Natural Science Foundation of Jiangsu Province (No. 13KJB430025), Environmental Science Research Project of Jiangsu Province (No. 2013004), the Natural Science Foundation of Jiangsu Province (No. BK20141172), Yancheng City Industrial Science and Technology Support Project (2013), and the Teaching Reform Project of Changzhou University (No. GJY2014068).

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Authors and Affiliations

  1. School of Chemical and Chemistry Engineering, Yancheng Institute of Technology, Yancheng, China

    Yong Dai, Bin Huan & Hai-Sheng Zhang

  2. College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China

    Yu-Cai He

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  1. Yong Dai
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Correspondence to Yong Dai.

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Dai, Y., Huan, B., Zhang, HS. et al. Effective Biotransformation of Ethyl 4-Chloro-3-Oxobutanoate into Ethyl (S)-4-Chloro-3-Hydroxybutanoate by Recombinant E. coli CCZU-T15 Whole Cells in [ChCl][Gly]–Water Media. Appl Biochem Biotechnol 181, 1347–1359 (2017). https://doi.org/10.1007/s12010-016-2288-0

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