Biosynthesis of ethyl (S)-4-chloro-3-hydroxybutanoate with an NADH-dependent reductase (ClCR) discovered by genome data mining using a modified colorimetric screening strategy.
An NADH-dependent reductase (ClCR) was discovered by genome data mining. After ClCR was overexpressed in E. coli BL21, recombinant E. coli CCZU-T15 with high reductase activity and excellent stereoselectivity for the reduction of ethyl 4-chloro-3-oxobutanoate (COBE) into ethyl (S)-4-chloro-3-hydroxybutanoate [(S)-CHBE] was screened using a modified high-throughput colorimetric screening strategy. After the reaction optimization, a highly stereoselective bioreduction of COBE into (S)-CHBE (>99% ee) with the resting cells of E. coli CCZU-T15 was successfully demonstrated in toluene-water (50:50, v/v) biphasic system. Biotransformation of 1000 mM COBE for 24 h in the biphasic system, (S)-CHBE (>99% ee) could be obtained in the high yield of 96.4%. Significantly, E. coli CCZU-T15 shows high potential in the industrial production of (S)-CHBE (>99% ee).
a Laboratory of Biocatalysis and Bioprocessing ; College of Pharmaceutical Engineeing and Life Sciences; Changzhou University , Changzhou , PR China.
Yu-Cai
YC
He
Dan-Ping
DP
Zhang
Yun
Y
Lu
Zhi-Cheng
ZC
Tao
Yun
Y
Ding
Li-Qun
LQ
Wang
Feng
F
Liu
D000090:Acetoacetates / Q000737:chemistry*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002087:Butyrates / Q000737:chemistry*; D057225:Data Mining; D004926:Escherichia coli; D057166:High-Throughput Screening Assays; D007218:Industrial Microbiology; D009243:NAD / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism
D000090:Acetoacetates; D001426:Bacterial Proteins; D002087:Butyrates; C089121:ClcR protein, bacteria; D015534:Trans-Activators; C121244:ethyl 4-chloro-3-hydroxybutanoate; D009243:NAD; C111982:ethyl 4-chloro-3-oxobutanoate
asymmetric biotransformation; ethyl (S)-4-chloro-3-hydroxybutanoate; ethyl 4-chloro-3-oxobutanoate; recombinant E. coli CCZU-T15; reductase