Discovery,characterization and mechanism of a Microbacterium esterase for key d-biotin chiral intermediate synthesis  

作　　者：Xinjia Li Haoran Yu Shengli Liu Baodi Ma Xiaomei Wu Xuesong Zheng Yi Xu 

机构地区：[1]School of Chemical and Environmental Engineering,Shanghai Institute of Technology,100 Haiquan Road,Shanghai 201418,China [2]Xianghu Laboratory,Hangzhou 311231,China [3]Institute of Bioengineering,College of Chemical and Biological Engineering,Zhejiang University,Hangzhou,Zhejiang 310027,China [4]Shandong Lonct Enzymes Co.,Ltd,Linyi 276400,China [5]School of Perfume and Aroma Technology,Shanghai Institute of Technology,100 Haiquan Road,Shanghai 201418,China

出　　处：《Bioresources and Bioprocessing》2024年第1期796-808,共13页生物资源与生物加工(英文)

基　　金：supported by the Shanghai Committee of Science and Technology(No.13430503400);Project of Leading Talents in Shandong Taishan Industry(Grant No.LJNY202019);the Scientific Research Foundation for the Returned Overseas Chinese Scholars,State Education Ministry(No.ZX2012-05);Innovation Program of Shanghai Municipal Education Commission(No.11YZ227).

摘　　要：Esterases are crucial biocatalysts in chiral compound synthesis.Herein,a novel esterase EstSIT01 belonging to family V was identified from Microbacterium chocolatum SIT101 through genome mining and phylogenetic analysis.EstSIT01 demonstrated remarkable efficiency in asymmetrically hydrolyzing meso-dimethyl ester[Dimethyl cis-1,3-Dibenzyl-2-imidazolidine-4,5-dicarboxyate],producing over 99%yield and 99%enantiomeric excess(e.e.)for(4S,5R)-monomethyl ester,a crucial chiral intermediate during the synthesis of d-biotin.Notably,the recombinant E.coli expressing EstSIT01 exhibited over 40-fold higher activity than that of the wild strain.EstSIT01 displays a preference for short-chain p-NP esters.The optimal temperature and pH were 45°C and 10.0,with Km and kcat values of 0.147 mmol/L and 5.808 s−1,respectively.Molecular docking and MD simulations suggest that the high stereoselectivity for meso-diester may attribute to the narrow entrance tunnel and unique binding pocket structure.Collectively,EstSIT01 holds great potential for preparing chiral carboxylic acids and esters.

关 键 词：Genome mining ESTERASE Stereoselective hydrolysis D-BIOTIN 

分 类 号：O62[理学—有机化学]