Engineered baker's yeast as whole-cell biocatalyst for one-pot stereo-selective conversion of amines to alcohols
(2014) In Microbial Cell Factories 13.- 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 omega-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... (More)
- 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 omega-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. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/4652855
- author
- Weber, Nora LU ; Gorwa-Grauslund, Marie-Francoise LU and Carlquist, Magnus LU
- organization
- publishing date
- 2014
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Transamination, Reduction, VAMT, Saccharomyces cerevisiae, Capsicum, chinense, Lactobacillus kefir, NADPH regeneration, Pyruvate, PLP, Kinetic resolution, Asymmetric synthesis
- in
- Microbial Cell Factories
- volume
- 13
- article number
- 118
- publisher
- BioMed Central (BMC)
- external identifiers
-
- wos:000340803400001
- scopus:84908390359
- pmid:25266107
- ISSN
- 1475-2859
- DOI
- 10.1186/s12934-014-0118-z
- language
- English
- LU publication?
- yes
- id
- db07a90f-fb5a-450b-81b9-27d6311c5a13 (old id 4652855)
- date added to LUP
- 2016-04-01 13:03:09
- date last changed
- 2022-03-13 21:50:21
@article{db07a90f-fb5a-450b-81b9-27d6311c5a13,
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 omega-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.}},
author = {{Weber, Nora and Gorwa-Grauslund, Marie-Francoise and Carlquist, Magnus}},
issn = {{1475-2859}},
keywords = {{Transamination; Reduction; VAMT; Saccharomyces cerevisiae; Capsicum; chinense; Lactobacillus kefir; NADPH regeneration; Pyruvate; PLP; Kinetic resolution; Asymmetric synthesis}},
language = {{eng}},
publisher = {{BioMed Central (BMC)}},
series = {{Microbial Cell Factories}},
title = {{Engineered baker's yeast as whole-cell biocatalyst for one-pot stereo-selective conversion of amines to alcohols}},
url = {{http://dx.doi.org/10.1186/s12934-014-0118-z}},
doi = {{10.1186/s12934-014-0118-z}},
volume = {{13}},
year = {{2014}},
}