A novel reductase from Candida albicans for the production of ethyl (S)-4-chloro-3-hydroxybutanoate.
A novel NADPH-dependent reductase (CaCR) from Candida albicans was cloned for the first time. It catalyzed asymmetric reduction to produce ethyl (S)-4-chloro-3-hydroxybutanoate ((S)-CHBE). It contained an open reading frame of 843 bp encoding 281 amino acids. When co-expressed with a glucose dehydrogenase in Escherichia coli, recombinant CaCR exhibited an activity of 5.7 U/mg with ethyl 4-chloro-3-oxobutanoate (COBE) as substrate. In the biocatalysis of COBE to (S)-CHBE, 1320 mM (S)-CHBE was obtained without extra NADP+/NADPH in a water/butyl acetate system, and the optical purity of the (S)-isomer was higher than 99% enantiomeric excess.
College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, PR China.
Mingdong
M
An
Ping
P
Cai
Ming
M
Yan
Ning
N
Hao
Shanshan
S
Wang
Huan
H
Liu
Yan
Y
Li
Lin
L
Xu
D000085:Acetates / Q000737:chemistry; D000090:Acetoacetates / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D055162:Biocatalysis; D002087:Butyrates / Q000138:chemical synthesis / Q000378:metabolism*; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005948:Glucose Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009249:NADP / Q000378:metabolism; D016366:Open Reading Frames; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D013237:Stereoisomerism; D014867:Water / Q000737:chemistry
D000085:Acetates; D000090:Acetoacetates; D002087:Butyrates; D029968:Escherichia coli Proteins; D005656:Fungal Proteins; C121244:ethyl 4-chloro-3-hydroxybutanoate; D014867:Water; C006848:butyl acetate; D009249:NADP; C111982:ethyl 4-chloro-3-oxobutanoate; D010088:Oxidoreductases; D005948:Glucose Dehydrogenases