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Efficient production of an ezetimibe intermediate using carbonyl reductase coupled with glucose dehydrogenase
Author(s) -
Zhang XiaoJian,
Zhou Rong,
Wu Di,
Tang YaQun,
Wang MengYing,
Liu ZhiQiang,
Zheng YuGuo
Publication year - 2020
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.3068
Subject(s) - cofactor , yield (engineering) , reductase , dehydrogenase , chemistry , substrate (aquarium) , nad+ kinase , biocatalysis , enantiomeric excess , nicotinamide adenine dinucleotide phosphate , biochemistry , ezetimibe , biosynthesis , catalysis , stereoselectivity , stereochemistry , enzyme , biology , enantioselective synthesis , reaction mechanism , materials science , cholesterol , ecology , oxidase test , metallurgy
Ezetimibe is a top‐selling hypolipidemic drug for the treatment of cardiovascular diseases. Biosynthesis of (4 S )‐3‐[(5 S )‐5‐(4‐fluorophenyl)‐5‐hydroxypentanoyl]‐4‐phenyl‐1,3‐oxazolidin‐2‐one (( S )‐ET‐5) using carbonyl reductase has shown advantages including high catalytic efficiency, excellent stereoselectivity, mild reaction conditions, and environmental friendness, and was considered as the key step for ezetimibe production. The regeneration efficiency of the cofactor, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) is one of the main restricted factor. Recombinant Escherichia coli strain (smCR125) coexpressing carbonyl reductase (CR125) and glucose dehydrogenase were successfully constructed and applied for the production of ( S )‐ET‐5 for the first time. Without extra addition of the coenzyme NADPH, the yield of 99.8% and the enantiomeric excess ( e.e. ) of 99.9% were achieved under ET‐4 concentration of 200 g/L. Using a substrate fed‐batch strategy, under the optimal conditions, the substrate ET‐4 concentration was increased to 250 g/L with the yield of 98.9% and the e.e. of 99.9% after 12 hr reaction. The space–time yield of 494.5 g L −1 d −1 and the space–time yield per gram biocatalyst of 24.7 g L −1 d −1 g −1 DCW were achieved, which were higher than ever reported for the biosynthesis of the ezetimibe intermediate.