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Construction of P450 scaffold biocatalysts for the biodegradation of five chloroanilines

文献类型： 外文期刊

作者： Yang, Ridong 1 ; Shoji, Osami 3 ; Lin, Yingwu 4 ; Wang, Fang 1 ; Che, Hongxia 2 ; Xu, Jiakun 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, State Key Lab Mariculture Biobreeding & Sustainabl, Lab Marine Drugs & Bioprod,Qingdao Marine Sci & Te, Qingdao 266071, Peoples R China

2.Qingdao Univ Sci & Technol, Coll Biol Engn, Shandong Prov Key Lab Biochem Engn, Qingdao 266042, Shandong, 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： P450BM3; Artificial metalloenzyme; Chloroanilines; Biodegradation

期刊名称：JOURNAL OF HAZARDOUS MATERIALS （ 影响因子：11.3； 五年影响因子：12.4 ）

ISSN： 0304-3894

年卷期： 2025 年 488 卷

页码：

收录情况： SCI

摘要： Chloroanilines represent a class of persistent and highly toxic environmental pollutants, posing significant challenges for green remediation strategies. While P450BM3 monooxygenases are renowned for their ability to catalyze the monooxidation of inert C-H bonds, costly NAD(P)H and complex electron transport systems required for P450BM3 catalysis limit their practical applications. This study pioneers the development of innovative artificial biocatalysts by strategically engineering the active site of P450BM3. Specifically, the substitution of the highly conserved threonine 268 with aspartic acid effectively induces peroxygenase activity, allowing for enhanced catalytic efficiency. Remarkably, the engineered P450BM3 mutants achieved degradation rates of 98.38-99.18 % for five chloroanilines (4-chloroaniline, 2-chloroaniline, 2,4-dichloroaniline, 3,4-dichloroaniline, and 3,5-dichloroaniline) in just 10-15 min, all without the need for NAD(P)H or dual-functional small molecules. Comprehensive degradation mechanism analysis via UPLC-MS corroborated the remarkable performance of these biocatalysts. This research not only demonstrates a novel approach for engineering P450 monooxygenases to exhibit peroxygenase activity but also significantly broadens their potential applications in synthetic chemistry and synthetic biology, paving the way for greener and more sustainable remediation technologies.

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