Bioproduction of enantiopure (r)- and (s)-2-phenylglycinol from styrene and renewable feedstocks via artificial enzyme cascade
AssigneeNational University of Singapore
InventorsZhi Li, Balaji Sundara Sekar, Benedict Ryan Lukito
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Abstract
Disclosed herein are methods of bioproduction of enantiomerically pure or enantiomerically enriched 2-phenylglycinol or a derivative thereof by multiple enzyme-catalyzed chemical transformations in a one-pot reaction system.
Claims27
A method for producing an enantiomerically pure or enantiomerically enriched (R)- or (S)-2-phenylglycinol or a derivative thereof using one or more recombinant microbial cells genetically engineered to overexpress multiple enzymes, which method comprises subjecting styrene or a derivative thereof to multiple enzyme-catalyzed chemical transformations in a one-pot reaction system.
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An isolated nucleic acid molecule encoding at least one heterologous catalytic enzyme selected from the group comprising:
- (a) an epoxidase for conducting an epoxidation reaction to form styrene oxide or a derivative thereof from styrene or a derivative thereof and an epoxide hydrolase for conducting a hydrolysis reaction on the epoxide to provide phenylethanediol or a derivative thereof, or an oxygenase for generating phenylethanediol or a derivative thereof from styrene or a derivative thereof by a dihydroxylation reaction:
- (b) an alcohol dehydrogenase or a secondary alcohol oxidase for generating 2-hydroxyacetophenone or a derivative thereof from phenylethanediol by an oxidation reaction:
- (c) a transaminase or an amine dehydrogenase for generating enantiomerically pure or enantiomerically enriched 2-phenylglycinol or a derivative thereof by a transamination reaction or a reductive amination from 2-hydroxyacetophenone or a derivative thereof;
- of (d) an ammonia lyase for generating trans-cinnamic acid or a derivative thereof from L-phenylalanine or a derivative thereof by a deamination reaction;
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- (e) a decarboxylase for generating styrene or a derivative thereof from trans-cinnamic acid or a derivative thereof in a decarboxylation reaction.
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Dependencies are inferred from the claim text.