数字农科院2.0

Synergistic Tuning of Conformational Dynamics, Electron Tunneling, and Substrate Positioning Enhances Electron Transfer in a P450 Chimera for Calcifediol Biosynthesis

文献类型： 外文期刊

作者： Liang, Ziqi;Song, Xitong;Cheng, Shuming;Shen, Yiwen;Zhang, Jie;Wang, Yaru;Luo, Huiying;Yao, Bin;Wang, Binju;Tu, Tao

作者机构：

关键词： calcifediol biocatalysis;conformational;dynamics modulation;electron transfer;multi-domain P450s engineering

期刊名称： ADVANCED SCIENCE

ISSN：

年卷期： 2026 年

页码：

收录情况： SCIE（2025版） ; ; EI（2025版）

摘要： Electron transfer (ET) efficiency dictates catalytic performance in multi-domain self-sufficient cytochrome P450s. Conventional engineering, however, predominantly focuses on localized optimization of either the active-site pocket or linker regions, overlooking inter-domain conformational transitions and ET chain integrity. Herein, we report a holistic ET-optimization strategy integrating conformational dynamics modulation, ET pathway engineering, and substrate positioning tuning, which was applied to enhance ET in a chimeric P450 VK1-CYP116B46-L21 (L21) for calcifediol biosynthesis. [2Fe-2S]-> Heme ET pathway engineering yielded variant L21-M2 (F346K/R354M), which decreased the conformational transition barrier by 4.5 kcal/mol and shortened the ET pathway by 4.05 & Aring;, leading to a 72-fold enhancement in ET rate. Heme domain engineering generated variant L21-M3 (P83A/A177M/K180F), which shortened the near-attack conformations distance to 4.19 & Aring; (from 4.62 & Aring;) and increased reactive conformation to 38 % (from 25 %). The pentuple variant L21-M5 combined both improvements, which demonstrated exceptional catalytic performance: an 8.2- fold higher catalytic efficiency, a coupling efficiency of 56.78 %, and a total turnover number (TTN) of 3222. In a semi-preparative-scale biotransformation, L21-M5 achieved 3.26 g/L production of calcifediol with 82 % conversion, underscoring its strong industrial potential. These results highlight the efficacy of the proposed ET-optimization strategy and provide a transferable workflow for engineering multi-domain redox biocatalysts.

分类号：

  • 相关文献

[1]ZmFdC2 Encoding a Ferredoxin Protein With C-Terminus Extension Is Indispensable for Maize Growth. Chen Yue,Zhong Deyi,Yang Xiu,Zhao Yonghui,Dai Liping,Zeng Dali,Wang Quan,Gao Lei,Li Shengben. 2021

[2]ZmFdC2 Encoding a Ferredoxin Protein With C-Terminus Extension Is Indispensable for Maize Growth. Chen Yue,Zhong Deyi,Yang Xiu,Zhao Yonghui,Dai Liping,Zeng Dali,Wang Quan,Gao Lei,Li Shengben. 2021

[3]Electron-rich Au nanocrystals/Co3O4 interface for enhanced electrochemical nitrate reduction into ammonia. Maolin Zhang,Kepeng Song,Chen Liu,Zedong Zhang,Wen Qing He,Hao Huang,Jialei Liu. 2023

[4]Fulvic acid more facilitated the soil electron transfer than humic acid. Side Yang,Kai Wang,Xin Yu,Yan Xu,Huike Ye,Mohan Bai,Lixia Zhao,Yang Sun,Xiaojing Li,Yongtao Li. 2024

[5]The role of semi-artificial photosynthetic systems in energy and environmental solutions: a critical review. Zhao, Shenggeng,Liu, Qiang,Li, Yeqing,Feng, Lu,Fu, Shanfei,Mazarji, Mahmoud,Pan, Junting. 2024

[6]How Biochar Addition Affects Denitrification and the Microbial Electron Transport System (ETSA): A Meta-Analysis Based on a Global Scale. Xiaolei Zhang,Qiwen Zhou,Lili Wang,Bo Wan,Qiannan Yang. 2024

[7]How Biochar Addition Affects Denitrification and the Microbial Electron Transport System (ETSA): A Meta-Analysis Based on a Global Scale. Xiaolei Zhang,Qiwen Zhou,Lili Wang,Bo Wan,Qiannan Yang. 2025

[8]Iron oxides accelerate soil electron transport to promote the degradation of organic pollutants: A review. Yu, Xin,Yang, Yuewei,Zhang, Yun,Sun, Jialu,Zhang, Xiaolin,Zhao, Xiaodong,Wang, Kai,Li, Tian,Li, Xiaojing. 2025

[9]Mn loading on montmorillonite surfaces for Cd stabilization: Insights from density functional theory calculations and surface complexation modeling. Meng Wang,Lei Yu,Jing Wang,Luyao Qin,Xiaoyi Sun,Jiaxiao Liu,Shibao Chen. 2025

[10]Accelerating electron transfer reduces CH4 and CO2 emissions in paddy soil. Yuewei Yang,Side Yang,Jialu Sun,Yun Zhang,Xin Yu,Penghui Li,Xiaolin Zhang,Xiaojing Li. 2025

[11]S–Mo conversion stimulates soil electron transfer and hydrocarbon removal for nearly one year. Jialu Sun,Xiaojuan Yi,Yuewei Yang,Yun Zhang,Xin Yu,Side Yang,Xiaolin Zhang,Ting Han,Xinhong Peng,Xiaojing Li. 2025

[12]Ferrihydrite redox enhanced biodegradation of petroleum hydrocarbons coupling with conversion of soil multi-elements by electron transfer. Xin Yu,Xiaolin Zhang,Pinpin Yang,Jialu Sun,Yuewei Yang,Xiaodong Zhao,Guang Yang,Penghui Li,Xiaojing Li. 2025

[13]Carbon-based mediators for extracellular electron transfer in environmental applications: a review. Side Yang,Xingping Chang,Xiaodong Zhao,Xiaolin Zhang,Kai Wang,Xiaojing Li,Yongtao Li. 2025

[14]Superoxide radical (·O₂⁻)–driven peroxymonosulfate activation via cation-deficient lanthanum ferrite perovskite oxides : Electronic structure modulation for high-efficiency estrogen degradation in dairy wastewater. Shiyu Lv,Tian Yuan,Xueyan Zhang,Menghan Feng,Yuan Luo,Weilin Fu,Kerong Fu,Yanli Luo,Feng Wang,Jixiu Wang. 2025

[15]Biochar loaded with Cu(I) to activate peroxyacetic acid for sulfamethoxazole removal: Effect of electron transfer and free radicals. Zeng, Chenyu,Ding, Anting,Chen, Xi,Ma, Yongfei,Lin, Hui,Ding, Han,Xie, Fuyu,Zang, Lu,Cui, Song,Li, Ping,Ma, Junwei,Zhang, Zulin. 2025

作者其他论文 更多>>