Yue Pan, Jinxiao Bao, Xingyi Zhang, Hui Ni, Yue Zhao, Fengdong Zhi, Bohuan Fang, Xiao He*, John Z. H. Zhang* and Lujia Zhang*
Aromatic aldehydes are important industrial raw materials mainly synthesized by anti Markovnikov (AM) oxidation of corresponding aromatic olefins. The AM product selectivity remains a big challenge. P450 aMOx is the first reported enzyme that could catalyze AM oxidation of aromatic olefins. Here, we reported a rational design strategy based on the “butterfly” model of the active site of P450 aMOx. Constrained molecular dynamic simulations and a binding energy analysis of key residuals combined with an experimental alanine scan were applied. As a result, the mutant A275G showed high AM selectivity of >99%. The results also proved that the “butterfly” model is an effective design strategy for enzymes.
