Discovery of a reductase-producing strain recombinant E. coli CCZU-A13 using colorimetric screening and its whole cell-catalyzed biosynthesis of ethyl (R)-4-chloro-3-hydroxybutanoate

An NADH-dependent reductase (SsCR) was discovered by genome data mining. After SsCR was overexpressed in E. coli BL21, recombinant E. coli CCZU-A13 with high reductase activity and excellent stereoselectivity for the reduction of ethyl 4-chloro-3-oxobutanoate (COBE) into ethyl ( R )-4-chloro-3-hydroxybutanoate (( R )-CHBE) was screened using one high-throughput colorimetric screening strategy. After the reaction optimization, a highly stereoselective bioreduction of COBE into ( R )-CHBE (>99% ee ) with the resting cells of E. coli CCZU-A13 was successfully demonstrated in n -butyl acetate–water (10:90, v/v) biphasic system. Biotransformation of 600   mM COBE for 8   h in the biphasic system, ( R )-CHBE (>99% ee ) could be obtained in the high yield of 100%. Moreover, the broad substrate specificity in the reduction of aliphatic and aromatic carbonyl compounds was also found. Significantly, E. coli CCZU-A13 shows high potential in the industrial production of ( R )-CHBE (>99% ee ) and its derivatives.