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


ABSTRACT: BACKGROUND: One-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. RESULTS: In this study, a heterologous enzymatic system consisting of ?-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. CONCLUSIONS: The 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.

SUBMITTER: Weber N 

PROVIDER: S-EPMC4423645 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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

Weber Nora N   Gorwa-Grauslund Marie M   Carlquist Magnus M  

Microbial cell factories 20140812


<h4>Background</h4>One-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.<h4>Results</h4>In this study, a heterologous enzymatic system consisting of ω-transaminase and ke  ...[more]

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