DataSheet1_Rational Design of P450 aMOx for Improving Anti-Markovnikov Selectivity Based on the “Butterfly” Model.docx

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.

芳香醛是一类重要的工业原料，其主要通过对应芳香烯烃的反马氏（anti-Markovnikov, AM）氧化反应合成。反马氏产物的选择性至今仍是一大挑战。P450 aMOx是首个被报道可催化芳香烯烃反马氏氧化反应的酶。本研究基于P450 aMOx活性位点的“蝴蝶”模型，提出了一套理性设计策略，采用约束分子动力学模拟、关键残基结合能分析，并结合丙氨酸扫描实验开展验证。结果显示，突变体A275G的反马氏产物选择性可达99%以上。该研究同时证实，“蝴蝶”模型是一种行之有效的酶理性设计策略。