Abstract
Biocatalytic reduction catalyzed by aldo-keto reductases (AKRs) is a valuable approach for asymmetric synthesis of chiral alcohols. In this study, four novel aldo-keto reductases with significant activity and stereoselectivity toward a variety of α-keto esters and halogen-substituted acetophenones were identified by genome mining. Through analysis of the crystal structure and multiple-sequence alignment of the starting AKR YvgN from Bacillus subtilis, residues F25 and W113 were proposed as the key positions that might control the stereoselectivity of YvgN. F25S and F25S/W113F variants of YvgN were able to improve its activity and stereoselectivity toward some α-keto ester compounds and halogen-substituted acetophenone derivatives. In addition, similar enhancement of catalytic activity and stereoselectivity was also found in the other three AKRs with corresponding mutations of starting YvgN.
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Funding
This work was financially supported by the 2018 Special Funds from the Central Treasury for the Reformation and Development of Local Areas: Development Platform for in vitro Diagnosis and Small Molecular Therapeutic Drugs, the Young Elite Scientists Sponsorship Program by CAST (2016QNRC001), National Natural Science Foundation of China (81602993), Science and Technology Research Projects from the Ministry of Education of the People’s Republic of China (213007A), and Project from the Department of Education of Liaoning Province.
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Zhang, W., Zhu, T., Li, H. et al. Key sites insight on the stereoselectivity of four mined aldo-keto reductases toward α-keto esters and halogen-substituted acetophenones. Appl Microbiol Biotechnol 103, 6119–6128 (2019). https://doi.org/10.1007/s00253-019-09932-7
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DOI: https://doi.org/10.1007/s00253-019-09932-7