Asymmetric synthesis of (S)-ethyl-4-chloro-3-hydroxy butanoate using a Saccharomyces cerevisiae reductase: enantioselectivity and enzyme-substrate docking studies.
Ethyl (S)-4-chloro-3-hydroxy butanoate (ECHB) is a building block for the synthesis of hypercholesterolemia drugs. In this study, various microbial reductases have been cloned and expressed in Escherichia coli. Their reductase activities toward ethyl-4-chloro oxobutanoate (ECOB) have been assayed. Amidst them, Baker's yeast YDL124W, YOR120W, and YOL151W reductases showed high activities. YDL124W produced (S)-ECHB exclusively, whereas YOR120W and YOL151W made (R)-form alcohol. The homology models and docking models with ECOB and NADPH elucidated their substrate specificities and enantioselectivities. A glucose dehydrogenase-coupling reaction was used as NADPH recycling system to perform continuously the reduction reaction. Recombinant E. coli cell co-expressing YDL124W and Bacillus subtilis glucose dehydrogenase produced (S)-ECHB exclusively.
Jung
Jihye
J
Division of Biotechnology, The Catholic University of Korea, Bucheon 420-743, Republic of Korea.
Park
Hyun Joo
HJ
Uhm
Ki-Nam
KN
Kim
Dooil
D
Kim
Hyung-Kwoun
HK
D001412:Bacillus subtilis / Q000378:metabolism; D002087:Butyrates / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D042843:Glucose 1-Dehydrogenase / Q000378:metabolism; D008956:Models, Chemical; D009249:NADP / Q000378:metabolism; D010088:Oxidoreductases / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D013237:Stereoisomerism; D013379:Substrate Specificity
D002087:Butyrates; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C121244:ethyl 4-chloro-3-hydroxybutanoate; D009249:NADP; D010088:Oxidoreductases; D042843:Glucose 1-Dehydrogenase