Title : Discovery, characterization and mechanism of a Microbacterium esterase for key d-biotin chiral intermediate synthesis - Li_2024_Bioresour.Bioprocess_11_59 |
Author(s) : Li X , Yu H , Liu S , Ma B , Wu X , Zheng X , Xu Y |
Ref : Bioresour Bioprocess , 11 :59 , 2024 |
Abstract :
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 degreesC and 10.0, with K(m) and k(cat) 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. |
PubMedSearch : Li_2024_Bioresour.Bioprocess_11_59 |
PubMedID: 38879848 |
Gene_locus related to this paper: 9mico-EstSIT01 , 9mico-a0a0m8mqm5 , 9mico-a0a0m9vm10 |
Gene_locus | 9mico-EstSIT01 9mico-a0a0m8mqm5 9mico-a0a0m9vm10 |
Li X, Yu H, Liu S, Ma B, Wu X, Zheng X, Xu Y (2024)
Discovery, characterization and mechanism of a Microbacterium esterase for key d-biotin chiral intermediate synthesis
Bioresour Bioprocess
11 :59
Li X, Yu H, Liu S, Ma B, Wu X, Zheng X, Xu Y (2024)
Bioresour Bioprocess
11 :59