A Novel NADH-Dependent Carbonyl Reductase from Kluyveromyces aestuarii and Comparison of NADH-Regeneration System for the Synthesis of Ethyl (S)-4-Chloro-3-hydroxybutanoate

Hiroaki YAMAMOTO; Kazuya MITSUHASHI; Norihiro KIMOTO; Akinobu MATSUYAMA; Nobuyoshi ESAKI; Yoshinori KOBAYASHI

doi:10.1271/bbb.68.638

Microbiology & Fermentation Technology Regular Paper

A Novel NADH-Dependent Carbonyl Reductase from Kluyveromyces aestuarii and Comparison of NADH-Regeneration System for the Synthesis of Ethyl (S)-4-Chloro-3-hydroxybutanoate

Hiroaki YAMAMOTO, Kazuya MITSUHASHI, Norihiro KIMOTO, Akinobu MATSUYAMA, Nobuyoshi ESAKI, Yoshinori KOBAYASHI

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Keywords: carbonyl reductase, ethyl (S)-4-chloro-3-hydroxybutanoate, formate dehydrogenase, Kluyveromyces aestuarii, NADH regeneration

JOURNAL FREE ACCESS

2004 Volume 68 Issue 3 Pages 638-649

DOI https://doi.org/10.1271/bbb.68.638

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Abstract

To compare NADH-regeneration systems for the synthesis of (S)-4-chloro-3-hydroxybutanoate (ECHB), a novel NADH-dependent carbonyl reductase (KaCR1), which reduced ethyl 4-chloroacetoacetate (ECAA) to form (S)-ECHB, was screened and purified from Kluyveromyces aestuarii and a gene encoding KaCR1 was cloned. Glucose dehydrogenase (GDH) and formate dehydrogenase (FDH) were compared as enzymes for NADH regeneration using Escherichia coli cells coexpressing each enzyme with KaCR1. E. coli cells coexpressing GDH produced 45.6 g/l of (S)-ECHB from 50 g/l of ECAA and E. coli cells coexpressing FDH, alternatively, produced only 19.0 g/l. The low productivity in the case of FDH was suggested to result from the low activity and instability of FDH.

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