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Regioselective aromatic O-demethylation with an artificial P450BM3/sugar alcohol oxidase peroxygenase system

文献类型： 外文期刊

作者： Zhao, Sijia 1 ; Lin, Yingwu 4 ; Wang, Jinghan 1 ; Li, Dong 1 ; Wang, Fang 1 ; Shoji, Osami 3 ; Xu, Jiakun 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, State Key Lab Mariculture Biobreeding & Sustainabl, Key Lab Sustainable Dev Polar Fisheries,Minist Agr, Qingdao 266071, Peoples R China

2.Ocean Univ China, Coll Food Sci & Engn, Qingdao Marine Sci & Technol Ctr, Lab Marine Drugs & Bioprod, Qingdao 266003, Peoples R China

3.Nagoya Univ, Grad Sch Sci, Dept Chem, Furo Cho,Chikusa Ku, Nagoya 4648602, Japan

4.Univ South China, Sch Chem & Chem Engn, Hengyang 421001, Peoples R China

关键词： Lignin; P450BM3; O-demethylation

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.5； 五年影响因子：8.7 ）

ISSN： 0141-8130

年卷期： 2025 年 309 卷

页码：

收录情况： SCI

摘要： The enzymatic demethylation of aromatic compounds presents a major challenge in the valorization of lignin. The main goal was to develop an efficient artificial peroxygenase system combining engineered P450BM3 with AldO (sugar alcohol oxidase) and DFSM (dual function small molecule) for the regioselective O-demethylation of lignin-derived aromatic ethers. P450BM3 serves as a versatile biocatalyst, and its engineered variants demonstrate expanded substrate promiscuity toward non-native substrates. AldO, served as the H2O2 in situ generation system. The DFSM, a rationally designed catalytic auxiliary, facilitates precise control of enzymatic reactions and enhances the efficiency of O-demethylation. We hypothesize that by combining P450BM3 with AldO and DFSM, we can better control the generation of H2O2 and direct the enzymatic system toward efficient O-demethylation. The engineered P450BM3 F87A/V78A/T268D/A328F mutant achieved a TON of 1895 +/- 4 for guaiacol, more than double that of the native P450BM3/H2O2 system (TON = 872 +/- 7). Moreover, the F87A/T268D mutant efficiently catalyzed double-demethylation of syringol, achieving the highest turnover number (TON) of 483 +/- 7. This DFSM-assisted P450BM3/AldO system represents a significant advancement in the biocatalytic degradation of lignin and offers a cost-effective and scalable alternative to traditional NADPH-dependent P450 monooxygenases. Our findings open new pathways for sustainable biotechnological applications in lignin valorization and aromatic compound catabolism.

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