Switching an Artificial P450 Peroxygenase into Peroxidase via Mechanism-Guided Protein Engineering

• DOI: 10.1021/acscatal.1c02698
• Publication Date: 2021-06-28T11:50:45Z
• Authors: Nana Ma; Wenhan Fang; Chuanfei Liu; Xiangquan Qin; Xiling Wang; Longyi Jin; Binju Wang; Zhiqi Cong
• Keywords: Protein Engineering; Guaiacol

Exploring the catalytic promiscuity of enzymes is a longstanding challenge and current topic interest. Our group previously modified cytochrome P450BM3 monooxygenase to perform peroxygenase activity with assistance from rationally designed dual-functional small molecule (DFSM). However, DFSM-facilitated P450-H2O2 system showed limited peroxidase activity. On basis mechanistic analysis possible competitive oxidation pathways, present work applies protein engineering strategy mutating redox-sensitive residues that enables achieve efficient The engineered exhibits one-electron toward various substrates, including guaiacol, 2,6-dimethoxyphenol, o-phenylenediamine, p-phenylenediamine. This attains best any P450 reported date rivals most natural peroxidases, suggesting significant potential for practical applications. provides insights strategies relevant expanding P450s through combining effects exogenous molecules.