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.
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.).
School of Chemical and Chemistry Engineering, Yancheng Institute of Technology, Yancheng, China. daiyong_yc@163.com.
Yong
http://orcid.org/0000-0001-7954-2969
Y
Dai
School of Chemical and Chemistry Engineering, Yancheng Institute of Technology, Yancheng, China.
Bin
B
Huan
School of Chemical and Chemistry Engineering, Yancheng Institute of Technology, Yancheng, China.
Hai-Sheng
HS
Zhang
College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China.
Yu-Cai
YC
He
D000090:Acetoacetates / Q000378:metabolism*; D001711:Biotransformation / Q000187:drug effects; D002087:Butyrates / Q000378:metabolism*; D002794:Choline / Q000737:chemistry; D003470:Culture Media / Q000737:chemistry* / Q000494:pharmacology*; D004274:DNA, Recombinant / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000166:cytology* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D019855:Ethylene Glycol / Q000737:chemistry; D005973:Glutamine / Q000737:chemistry; D011136:Polysorbates / Q000737:chemistry; D014867:Water / Q000737:chemistry
D000090:Acetoacetates; D002087:Butyrates; D003470:Culture Media; D004274:DNA, Recombinant; D011136:Polysorbates; C121244:ethyl 4-chloro-3-hydroxybutanoate; D014867:Water; D005973:Glutamine; C111982:ethyl 4-chloro-3-oxobutanoate; D019855:Ethylene Glycol; D002794:Choline
Choline chloride; E. coli CCZU-T15; Ethyl (S)-4-chloro-3-hydroxybutanoate; Ethyl 4-chloro-3-oxobutanoate; Tween-80