Chiral alcohol production by NADH-dependent phenylacetaldehyde reductase coupled with in situ regeneration of NADH.
Phenylacetaldehyde reductase (PAR) produced by styrene-assimilating Corynebacterium strain ST-10 was used to synthesize chiral alcohols. This enzyme with a broad substrate range reduced various prochiral aromatic ketones and beta-ketoesters to yield optically active secondary alcohols with an enantiomeric purity of more than 98% enantiomeric excess (e.e.). The Escherichia coli recombinant cells which expressed the par gene could efficiently produce important pharmaceutical intermediates; (R)-2-chloro-1-(3-chlorophenyl)ethanol (28 mg.mL-1) from m-chlorophenacyl chloride, ethyl (R)-4-chloro-3-hydroxy butanoate) (28 mg.mL-1) from ethyl 4-chloro-3-oxobutanoate and (S)-N-tert-butoxycarbonyl(Boc)-3-pyrrolidinol from N-Boc-3-pyrrolidinone (51 mg.mL-1), with more than 86% yields. The high yields were due to the fact that PAR could concomitantly reproduce NADH in the presence of 3-7% (v/v) 2-propanol in the reaction mixture. This biocatalytic process provided one of the best asymmetric reductions ever reported.
Itoh
Nobuya
N
Biotechnology research center, Toyama Prefectural University, Japan. itoh@pu-toyama.ac.jp
Matsuda
Michiko
M
Mabuchi
Mariko
M
Dairi
Tohru
T
Wang
Jincun
J
D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D000438:Alcohols / Q000378:metabolism*; D002384:Catalysis; D007700:Kinetics; D009243:NAD / Q000378:metabolism*; D013237:Stereoisomerism; D013379:Substrate Specificity
D000438:Alcohols; D009243:NAD; D000429:Alcohol Oxidoreductases; C110191:phenylacetaldehyde reductase