Engineered baker’s yeast as whole-cell biocatalyst for one-pot stereo-selective conversion of amines to alcohols

Microbial Cell Factories

Research

Marie Gorwa-Grauslund¹  Nora Weber¹  Magnus Carlquist¹ 

[1] Division of Applied Microbiology, Department of Chemistry, Lund University, SE-22100, Lund, Sweden;

关键词: Transamination;    Reduction;    VAMT;    Saccharomyces cerevisiae;    Capsicum chinense;    Lactobacillus kefir;    NADPH regeneration;    Pyruvate;    PLP;    Kinetic resolution;    Asymmetric synthesis;   

DOI  :  10.1186/s12934-014-0118-z

 received in 2014-06-04, accepted in 2014-08-02,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundOne-pot multi-step biocatalysis is advantageous over step-by-step synthesis as it reduces the number of process operation units, leading to significant process intensification. Whole-cell biocatalysis with metabolically active cells is especially valuable since all enzymes can be co-expressed in the cell whose metabolism can be exploited for supply of co-substrates and co-factors.ResultsIn this study, a heterologous enzymatic system consisting of ω-transaminase and ketone reductase was introduced in Saccharomyces cerevisiae, and evaluated for one-pot stereo-selective conversion of amines to alcohols. The system was applied for simultaneous kinetic resolution of racemic 1-phenylethylamine to (R)-1-phenylethylamine and reduction of the ketone intermediate to (R)-1-phenylethanol. Glucose was used as sole co-substrate for both the supply of amine acceptor and the regeneration of NADPH in the reduction step.ConclusionsThe whole-cell biocatalyst was shown to sustain transaminase-reductase-catalyzed enantioselective conversion of amines to alcohols with glucose as co-substrate. The transamination catalyzed by recombinant vanillin aminotransferase from Capsicum chinense proved to be the rate-limiting step as a three-fold increase in transaminase gene copy number led to a two-fold increased conversion. The (R)-selective NADPH-dependent alcohol dehydrogenase from Lactobacillus kefir proved to be efficient in catalyzing the reduction of the acetophenone generated in the transamination reaction.

【 授权许可】

Unknown   
© Weber et al.; licensee BioMed Central Ltd. 2014. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.