Zhang Chun, Liu Ping-Xian, Huang Lu-Yi, Wei Si-Ping, Wang Li, Yang Sheng-Yong, Yu Xiao-Qi, Pu Lin, Wang Qin
Department of Medicinal Chemistry, Southwest Medical University, No. 319, Zhongshan Road, Luzhou, Sichuan, 646000, P. R. China.
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, No.17 People's South Road, Chengdu, Sichuan, 610041, P. R. China.
Chemistry. 2016 Jul 25;22(31):10969-75. doi: 10.1002/chem.201601176. Epub 2016 Jun 30.
P450 119 peroxygenase and its site-directed mutants are discovered to catalyze the enantioselective epoxidation of methyl-substituted styrenes. Two new site-directed P450 119 mutants, namely T213Y and T213M, which were designed to improve the enantioselectivity and activity for the epoxidation of styrene and its methyl substituted derivatives, were studied. The T213M mutant is found to be the first engineered P450 peroxygenase that shows highly enantioselective epoxidation of cis-β-methylstyrenes, with up to 91 % ee. Molecular modeling studies provide insights into the different catalytic activity of the T213M mutant and the T213Y mutant in the epoxidation of cis-β-methylstyrene. The results of the calculations also contribute to a better understanding of the substrate specificity and configuration control for the regio- and stereoselective peroxygenation catalyzed by the T213M mutant.
发现细胞色素P450 119过氧合酶及其定点突变体可催化甲基取代苯乙烯的对映选择性环氧化反应。研究了两个新的细胞色素P450 119定点突变体，即T213Y和T213M，它们旨在提高苯乙烯及其甲基取代衍生物环氧化反应的对映选择性和活性。发现T213M突变体是首个对顺式-β-甲基苯乙烯表现出高对映选择性环氧化反应的工程化细胞色素P450过氧合酶，对映体过量值高达91%。分子模拟研究揭示了T213M突变体和T213Y突变体在顺式-β-甲基苯乙烯环氧化反应中不同的催化活性。计算结果也有助于更好地理解T213M突变体催化的区域和立体选择性过氧化反应的底物特异性和构型控制。
