Synthesis of ethyl ( S)-4-chloro-3-hydroxybutanoate using fabG-homologues.
This paper is a report on the successful application of bioinformatics to enzyme screening. The synthesis of ethyl ( S)-4-chloro-3-hydroxybutanoate (ECHB) by asymmetric reduction of ethyl 4-chloroacetoacetate (ECAA) using fabG-homologues was studied. beta-Ketoacyl-acyl carrier protein reductases from both Escherichia coli and Bacillus subtilis, which are components of type II fatty acid synthase, could reduce ECAA to ( S)-ECHB with 94-98% ee. Furthermore, acetoacetyl-CoA reductases (ARs) from both Ralstonia eutropha and Zoogloea ramigera, whose genes are significantly similar to fabG genes and play a physiological role in the biosynthesis of poly-beta-3-hydroxybutyrate, could also catalyze the asymmetric reduction of ECAA to ( S)-ECHB with >99% ee. ( S)-ECHB was synthesized to 48.7 g/l with an optical purity of 99.8% ee, using recombinant E. coli cells coexpressing AR from R. eutropha and glucose dehydrogenase from B. subtilis for the regeneration of NADPH.
Yamamoto
H
H
Life Science Development Center, CPI Company, Tsukuba Research Center, Daicel Chemical Industries, Ltd, 27 Miyukigaoka, Tsukuba 305-0841, Japan. h-yamamoto@daicel.co.jp
Matsuyama
A
A
Kobayashi
Y
Y
D064431:3-Oxoacyl-(Acyl-Carrier-Protein) Reductase; D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D020562:Betaproteobacteria / Q000201:enzymology / Q000235:genetics; D002087:Butyrates / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D042843:Glucose 1-Dehydrogenase; D005948:Glucose Dehydrogenases / Q000235:genetics / Q000378:metabolism; D006885:Hydroxybutyrates / Q000378:metabolism; D010957:Plasmids; D011091:Polyesters / Q000378:metabolism; D013379:Substrate Specificity; D000094122:Polyhydroxybutyrates
D064431:3-Oxoacyl-(Acyl-Carrier-Protein) Reductase; D000429:Alcohol Oxidoreductases; D002087:Butyrates; D042843:Glucose 1-Dehydrogenase; D005948:Glucose Dehydrogenases; D006885:Hydroxybutyrates; D011091:Polyesters; C121244:ethyl 4-chloro-3-hydroxybutanoate; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C023744:acetoacetyl-CoA reductase