数字农科院2.0

Efficient C25-Hydroxylation of Vitamin D3 Utilizing an Artificial Self-Sufficient Whole-Cell Cytochrome P450 Biocatalyst

文献类型： 外文期刊

作者： Liang, Ziqi;Zhou, Qiao;Li, Yicheng;Liu, Xiaoqing;Shen, Yiwen;Tian, Jian;Wang, Xiaolu;Qin, Xing;Wang, Yuan;Luo, Huiying;Yang, Xiaojun;Yao, Bin;Tu, Tao

作者机构：

关键词： 25-hydroxyvitamin D-3;P450s;self-sufficientP450s;local energetic frustration;linker engineering

期刊名称： JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY

ISSN： 0021-8561

年卷期： 2025 年

页码：

收录情况： SCIE（2025版） ; ; EI（2025版）

摘要： Cytochrome P450 enzymes (P450s) are promising candidates for the biosynthesis of 25-hydroxyvitamin D-3 (25(OH)VD3). However, their industrial application is limited by challenges, such as low stability, inefficient catalysis, and uncoupling reactions. The construction of self-sufficient P450s offers a strategic solution to these limitations, but requires linker optimization to regulate interdomain conformational dynamics. In this study, we integrated whole-cell biocatalyst screening with systematic optimization of reaction conditions, including cosolvents, cell concentrations, and plasmid selection, to enhance catalytic performance. Under optimized conditions, the heme domain Vdh-K1 achieved a 91.6% conversion efficiency and was subsequently selected for chimeric enzyme assembly. By employing local energetic frustration analysis to evaluate protein flexibility and allosteric dynamics, we identified chimeric P450 variants with highly frustrated linkers. The optimal variant, VK1-CYP116B46-L21, exhibited improved thermostability, catalytic activity, and coupling efficiency, achieving a yield of 4.89 mM (1.96 g/L) 25(OH)VD3 in Escherichia coli whole-cell catalysis-the highest reported yield to date. This work underscores the utility of computational frustration analysis in refining linker dynamics for multidomain enzymes and establishes a scalable, cost-effective framework to advance P450s systems for industrial biosynthesis of high-value compounds.

分类号：

  • 相关文献

[1]CYP6DW3 Metabolizes Imidacloprid to Imidacloprid-urea in Whitefly (Bemisia tabaci). Xue, Hu,Fu, Buli,Huang, Mingjiao,He, Chao,Liang, Jinjin,Yang, Jing,Wei, Xuegao,Liu, Shaonan,Du, Tianhua,Ji, Yao,Yin, Cheng,Gong, Peipan,Hu, JinYu,Du, He,Zhang, Rong,Wang, Chao,Khajehali, Jahangir,Su, Qi,Yang, Xin,Zhang, Youjun. 2023

[2]Pro-197-Ser Mutation and Cytochrome P450-Mediated Metabolism Conferring Resistance to Flucarbazone-Sodium in Bromus japonicus. Yuning Lan,Xinxin Zhou,Shenyuan Lin,Yi Cao,Shouhui Wei,Hongjuan Huang,Wenyu Li,Zhaofeng Huang. 2022

[3]Cytochrome P450 CYP6EM1 confers resistance to thiamethoxam in the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) via detoxification metabolism. Mingjiao Huang,Peipan Gong,Cheng Yin,Jing Yang,Shaonan Liu,Buli Fu,Xuegao Wei,Jinjin Liang,Hu Xue,Chao He,Tianhua Du,Chao Wang,Yao Ji,Jin Yu Hu,Rong Zhang,Natalia A. Belyakova,Youjun Zhang,Xin Yang. 2025

[4]Expression of Echinochloa oryzoides CYP71A1 and GSTU23 catalyzes the metabolism of thiobencarb, thereby conferring resistance. Jinhui Li,Yi Zhang,Longlong Li,Shouhui Wei,Zhaofeng Huang,Lu Chen,Hongjuan Huang. 2025

作者其他论文 更多>>