Stereospecific Reduction of Methyl O-Chlorobenzoylformate at 300 g/L-1 without Additional Cofactor using a Carbonyl Reductase Mined from Candida glabrata

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논문

Stereospecific Reduction of Methyl O-Chlorobenzoylformate at 300 g/L-1 without Additional Cofactor using a Carbonyl Reductase Mined from Candida glabrata

학술지

Advanced synthesis & catalysis

저자명

Ma, Hongmin; Yang, Linlin; Ni, Yan; Zhang, Jie; Li, Chun‐ Xiu; Zheng, Gao‐ Wei; Yang, Huaiyu; Xu, Jian‐ He

초록

AbstractIn order to search for oxidoreductases suitable for the preparation of methyl (R)‐o‐chloromandelate [(R)‐CMM], the key intermediate for clopidogrel, the homologous proteins of Gre2p were expressed in Escherichia coli, among which CgKR1 showed the most satisfactory activity and stereoselectivity towards methyl o‐chlorobenzoylformate (CBFM). Using the crude enzyme of CgKR1 and glucose dehydrogenase (GDH), as much as 300 g⋅L−1 of CBFM was almost stoichiometrically converted to (R)‐CMM with excellent enantiomeric excess (98.7% ee). More importantly, the reaction could be performed without external addition of an expensive cofactor. The substrate profile indicates that keto esters serve as the most suitable substrate, which was confirmed by gram‐scale preparations. Homology modeling and docking analysis revealed the molecular basis for the high stereoselectivity of CgKR1. These demonstrate not only the feasibility of in silico mining of novel enzymes based on sequence homology but also the applicability of this new reductase for the practical production of optically active (R)‐CMM.

발행연도

2012

발행기관

WILEY‐VCH Verlag

ISSN

1615-4150

ISSN

1615-4169

권

354

호

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