A novel insight of enhancing the hydrogen peroxide tolerance of unspecific peroxygenase from Daldinia caldariorum based on structure  

作　　者：Tiantian Li Ruochen Jin Bin Wu Dongming Lan Yunjian Ma Yonghua Wang 

机构地区：[1]School of Food Science and Engineering,South China University of Technology,Guangzhou 510640,China [2]School of Bioscience and Bioengineering,South China University of Technology,Guangzhou 510006,China [3]Guangdong Youmei Institute of Intelligent Bio-manufacturing Co.,Ltd.,Foshan 528200,China

出　　处：《Chinese Chemical Letters》2024年第4期426-430,共5页中国化学快报（英文版）

基　　金：supported by the National Natural Science Foundation of China (No.32001633);the Key Program of Natural Science Foundation of China (No.31930084);Guangzhou Science and technology planning project (No.202102020370)。

摘　　要：Unspecific peroxygenases(UPOs, EC 1.11.2.1) is a kind of thioheme enzyme capable of catalyzing various oxidations of inert C–H bonds using H_(2)O_(2) as an oxygen donor without cofactors. However, the enhancement of the H_(2)O_(2) tolerance of UPOs is always challenging. In this study, the A161C mutant of r Dca UPO,which originates from Daldinia caldariorum, was found to be highly H_(2)O_(2)-resistant. Compared with the wild type, the mutant r Dca UPO-A161C showed a 10-h prolonged half-life and a 64% improved enzyme activity when incubated in 10 mmol/L H_(2)O_(2). The crystal structure analysis at 1.47 A showed that r Dca UPOA161C exhibited 10 α-helixes(cyan) and a series of ordered rings, forming a single asymmetric spherical structure. The two conserved domains near heme formed an active site with the catalytic PCP and EHD regions(Glu86, His87, Asp88 residues). The H_(2)O_(2) tolerance of r Dca UPO-A161C was preliminarily explored by comparing its structure with the wild type. Notably, r Dca UPO-A161C showed significantly higher catalytic efficiency than the wild type for the production of hydroxyl fatty acids. This study is anticipated to provide an insight into the structure-function relationship and expand potential applications of UPOs.

关 键 词：Unspecific peroxygenase Molecular modification Hydrogen peroxide stability Crystal structure Hydroxyl fatty acids 

分 类 号：TQ426.97[化学工程]