摘要：

By genome data mining, a carbonyl reductase tool box was designed and developed for chiral alcohol synthesis. On the basis of systematic comparison of the specific activity and substrate tolerance toward α-chloroacetophenone among reductases in this tool box, KtCR, a highly substrate-/product-tolerant carbonyl reductase from Kluyveromyces thermotolerans, was identified. The reduction of a series of substituted aryl ketones was investigated using this newly mined biocatalyst. Almost all of the ketones tested were asymmetrically reduced into corresponding chiral alcohols in 99% ee. Substrates with substituents adjacent to the carbonyl group or those with substituents on the para position of the phenyl ring were easier to reduce. For α-choloacetophenone as a representative substrate, as much as 154 g/L (1.0 M) of the substrate was asymmetrically reduced within merely 12 h by lyophilized cells of Escherichia coli/pET28-KtCR, resulting in an isolated yield of 92%, an enantiopurity of >99% ee, and a total turno...

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