PMID:12655455
| Citation |
Yamamoto, H, Matsuyama, A and Kobayashi, Y (2003) Synthesis of ethyl ( S)-4-chloro-3-hydroxybutanoate using fabG-homologues. Appl. Microbiol. Biotechnol. 61:133-9 |
|---|---|
| Abstract |
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. |
| Links |
PubMed Online version:10.1007/s00253-002-1188-0 |
| Keywords |
Alcohol Oxidoreductases/genetics; Alcohol Oxidoreductases/metabolism; Bacillus subtilis/enzymology; Bacillus subtilis/genetics; Betaproteobacteria/enzymology; Betaproteobacteria/genetics; Butyrates/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Glucose 1-Dehydrogenase; Glucose Dehydrogenases/genetics; Glucose Dehydrogenases/metabolism; Hydroxybutyrates/metabolism; Plasmids; Polyesters/metabolism; Substrate Specificity |
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