文献类型：会议论文

中文题名：Expending biocatalyst tool-box: Genome mining for robust carbonyl reductases

作者：Guo-Chao Xu[1];Hui-Lei Yu[1];Jian-He Xu[1];

机构：[1]Laboratory of Biocatalysis and Synthetic Biotechnology, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, People''s Republic of China;

会议论文集：2012年全国博士生学术论坛——发酵工程论文集

会议日期：20120813

会议地点：上海

主办单位：教育部学位管理与研究生教育司,国务院学位办

语种：英文

中文关键词：Carbonyl;reductase;Asymmetric;hydrogen;transfer;Aryl;halohydrin;Kluyveromyces;thermotolerans;Genome;data;mining;Substrate/product;tolerance

摘要：By genome data mining, a carbonyl reductase tool-box was designed and developed for chiral alcohol synthesis.Based on systematic comparison of the specific activity and substrate tolerance towards 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 para-position of the phenyl ring were easier to be reduced.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 E.coli/pET28-KtCR, resulting in an isolated yield of 92%, an enantiopurity of >99% ee, and a total turnover number of 5000 which was five times higher than the highest record reported so far.These results indicate the great potential of KtCR in practical synthesis of valuable aryl halohydrins as versatile chiral synthons.