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Enantioselective synthesis of enantiopure chiral alcohols using carbonyl reductases screened from Yarrowia lipolytica.
J Appl Microbiol 2019; 126(1):127-137JA

Abstract

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, YaCRI and YaCRII, 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 YaCRI and YaCRII 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 BL21 (DE3)/pET-28a-yacrII 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.

Authors+Show Affiliations

Laboratory of Medicinal Chemistry and Molecular Diagnosis of the Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding, China.Laboratory of Medicinal Chemistry and Molecular Diagnosis of the Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding, China.Laboratory of Medicinal Chemistry and Molecular Diagnosis of the Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding, China.Laboratory of Medicinal Chemistry and Molecular Diagnosis of the Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding, China.Laboratory of Medicinal Chemistry and Molecular Diagnosis of the Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding, China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30291666

Citation

Zhang, H-L, et al. "Enantioselective Synthesis of Enantiopure Chiral Alcohols Using Carbonyl Reductases Screened From Yarrowia Lipolytica." Journal of Applied Microbiology, vol. 126, no. 1, 2019, pp. 127-137.
Zhang HL, Zhang C, Pei CH, et al. Enantioselective synthesis of enantiopure chiral alcohols using carbonyl reductases screened from Yarrowia lipolytica. J Appl Microbiol. 2019;126(1):127-137.
Zhang, H. L., Zhang, C., Pei, C. H., Han, M. N., & Li, W. (2019). Enantioselective synthesis of enantiopure chiral alcohols using carbonyl reductases screened from Yarrowia lipolytica. Journal of Applied Microbiology, 126(1), pp. 127-137. doi:10.1111/jam.14125.
Zhang HL, et al. Enantioselective Synthesis of Enantiopure Chiral Alcohols Using Carbonyl Reductases Screened From Yarrowia Lipolytica. J Appl Microbiol. 2019;126(1):127-137. PubMed PMID: 30291666.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enantioselective synthesis of enantiopure chiral alcohols using carbonyl reductases screened from Yarrowia lipolytica. AU - Zhang,H-L, AU - Zhang,C, AU - Pei,C-H, AU - Han,M-N, AU - Li,W, Y1 - 2018/10/29/ PY - 2018/03/30/received PY - 2018/09/12/revised PY - 2018/09/25/accepted PY - 2018/10/7/pubmed PY - 2018/10/7/medline PY - 2018/10/7/entrez KW - Yarrowia lipolytica KW - carbonyl reductase KW - chiral alcohols KW - stereoselective reduction KW - two-phase system SP - 127 EP - 137 JF - Journal of applied microbiology JO - J. Appl. Microbiol. VL - 126 IS - 1 N2 - 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, YaCRI and YaCRII, 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 YaCRI and YaCRII 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 BL21 (DE3)/pET-28a-yacrII 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. SN - 1365-2672 UR - https://www.unboundmedicine.com/medline/citation/30291666/Enantioselective_synthesis_of_enantiopure_chiral_alcohols_using_carbonyl_reductases_screened_from_Yarrowia_lipolytica_ L2 - https://doi.org/10.1111/jam.14125 DB - PRIME DP - Unbound Medicine ER -