Development of a substrate-coupled biocatalytic process driven by an NADPH-dependent sorbose reductase from Candida albicans for the asymmetric reduction of ethyl 4-chloro-3-oxobutanoate.
A substrate-coupled biocatalytic process was developed based on the reactions catalyzed by an NADPH-dependent sorbose reductase (SOU1) from Candida albicans in which ethyl 4-chloro-3-oxobutanoate (COBE) was reduced to (S)-4-chloro-3-hydroxybutanoate [(S)-CHBE], while NADPH was regenerated by the same enzyme via oxidation of sugar alcohols. (S)-CHBE yields of 1,140, 1,150, and 780 mM were obtained from 1,220 mM COBE when sorbitol, mannitol, and xylitol were used as co-substrates, respectively. Optimization of COBE and sorbitol proportions resulted in a maximum yield of (S)-CHBE (2,340 mM) from 2,500 mM COBE, and the enantiomeric excess was 99.6 %. The substrate-coupled system driven by SOU1 maintained a stable pH and a robust intracellular NADPH circulation; thus, pH adjustment and addition of extra coenzymes were unnecessary.
State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, People's Republic of China. tsaiping821218@gmail.com
Ping
P
Cai
Mingdong
M
An
Lin
L
Xu
Sheng
S
Xu
Ning
N
Hao
Yan
Y
Li
Kai
K
Guo
Ming
M
Yan
D000090:Acetoacetates / Q000378:metabolism*; D002087:Butyrates / Q000378:metabolism*; D002176:Candida albicans / Q000201:enzymology*; D003067:Coenzymes / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D009249:NADP / Q000378:metabolism*; D010084:Oxidation-Reduction; D013401:Sugar Alcohol Dehydrogenases / Q000378:metabolism*; D013402:Sugar Alcohols / Q000378:metabolism
D000090:Acetoacetates; D002087:Butyrates; D003067:Coenzymes; D013402:Sugar Alcohols; C121244:ethyl 4-chloro-3-hydroxybutanoate; D009249:NADP; C111982:ethyl 4-chloro-3-oxobutanoate; D013401:Sugar Alcohol Dehydrogenases; C506437:sorbose reductase