Protein Engineering of an Artificial P450BM3 Peroxygenase System Enables Highly Selective O-Demethylation of Lignin Monomers

文献类型： 外文期刊

第一作者： Li, Maosheng

作者： Li, Maosheng;Li, Maosheng;Miao, Hengmin;Li, Yanqing;Wang, Fang;Xu, Jiakun

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关键词： protein engineering; peroxygenase; lignin monomers; dual-functional small molecule; O-demethylation

期刊名称：MOLECULES （ 影响因子：4.927； 五年影响因子：5.11 ）

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年卷期： 2022 年 27 卷 10 期

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收录情况： SCI

摘要： The O-demethylation of lignin monomers, which has drawn substantial attention recently, is critical for the formation of phenols from aromatic ethers. The P450BM3 peroxygenase system was recently found to enable the O-demethylation of different aromatic ethers with the assistance of dual-functional small molecules (DFSM), but these prepared mutants only have either moderate O-demethylation activity or moderate selectivity, which hinders their further application. In this study, we improve the system by introducing different amino acids into the active site of P450BM3, and these amino acids with different side chains impacted the catalytic ability of enzymes due to their differences in size, polarity, and hydrophobicity. Among the prepared mutants, the combination of V78A/F87A/T268I/A264G and Im-C6-Phe efficiently catalyzed the O-demethylation of guaiacol (TON = 839) with 100% selectivity. Compared with NADPH-dependent systems, we offer an economical and practical bioconversion avenue.

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