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Enantioselective synthesis of enantiopure chiral alcohols using carbonyl reductases screened from Yarrowia lipolytica
Author(s) -
Zhang H.L.,
Zhang C.,
Pei C.H.,
Han M.N.,
Li W.
Publication year - 2019
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.14125
Subject(s) - yarrowia , enantiopure drug , escherichia coli , enantioselective synthesis , biocatalysis , reductase , chemistry , substrate (aquarium) , yeast , stereochemistry , biochemistry , enzyme , biology , catalysis , gene , reaction mechanism , ecology
Aims We aimed to explore Yarrowia lipolytica carbonyl reductases as effective biocatalysts and to develop efficient asymmetric reduction systems for chiral alcohol synthesis. Methods and Results Yarrowia lipolytica carbonyl reductase genes were obtained via homologous sequence amplification strategy. Two carbonyl reductases, Ya CRI and Ya CRII , were identified and characterized, and used to catalyse the conversion of 2‐hydroxyacetophenone (2‐ HAP ) to optically pure (S)‐1‐phenyl‐1,2‐ethanediol. Enzymatic assays revealed that Ya CRI and Ya CRII exhibited specific activities of 6·96 U mg −1 (99·8% e.e .) and 7·85 U mg −1 (99·9% e.e .), respectively, and showed moderate heat resistance at 40–50°C and acid tolerance at pH 5·0–6·0. An efficient whole‐cell two‐phase system was established using reductase‐expressing recombinant Escherichia coli . The conversion of 2‐ HAP (20·0 g l −1 ) conversion with the solvent of dibutyl phthalate was approximately 70‐fold higher than in water. Furthermore, the two recombinant E. coli displayed biocatalyst activity and enantioselectivity towards several different carbonyl compounds, and E. coli BL 21 ( DE 3)/ pET ‐28a‐ yacr II showed a broad substrate spectrum. Conclusions A new whole‐cell recombinant E. coli ‐based bioreduction system for enantiopure alcohol synthesis with high enantioselectivity at high substrate concentrations was developed. Significance and Impact of the Study We proposed a promising approach for the efficient preparation of enantiopure chiral alcohols.