Hongxun Gao, Yakun Fan, Xuwei He, Xiaogang Peng, Zhen Li, Yanxin Zheng, Shengbiao Ji, Longwu Ye, Aitao Li*, Binju Wang* and Jing Zhao*,
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引用次数: 0
Abstract
The P450 monooxygenase TleB (CYP107E48) catalyzes intramolecular C–S bond formation in a thiol-containing substrate, yielding two sulfur-containing indolactam derivatives (P1 and P2). However, the key sites influencing TleB’s product selectivity and the molecular mechanisms underlying the selective C–S bond formation are not fully understood. To address this, we created an artificial self-sufficient P450, TleB-CYP116B46, by fusing TleB with the reductase domain of CYP116B46. Structure-guided engineering of TleB-CYP116B46 generates variant L85G with 99% selectivity for P1 and variant I282L/Q387L/I234F with 95% selectivity for P2. Exploring TleB homologues and generating corresponding mutants elucidate the identified sites’ crucial role in product selectivity. Computational studies suggest a diradical mechanism for C–S bond formation for both P1 and P2 products. Intriguingly, we found that the substrate radical could undergo conformational changes in both the S–H and indole groups. The L85G variant facilitates the conformational switch of the indole radical group, thereby leading to the selective C–S bond formation for the P1 product. By contrast, the I282L/Q387L/I234F variant barricades the conformational switch of the indole radical group, affording the P2 product. Our simulations highlight that the protein environment can dictate the dynamics and positioning of the substrate radical, thereby leading to the selective C–S bond formation in P450s.
期刊介绍:
ACS Catalysis is an esteemed journal that publishes original research in the fields of heterogeneous catalysis, molecular catalysis, and biocatalysis. It offers broad coverage across diverse areas such as life sciences, organometallics and synthesis, photochemistry and electrochemistry, drug discovery and synthesis, materials science, environmental protection, polymer discovery and synthesis, and energy and fuels.
The scope of the journal is to showcase innovative work in various aspects of catalysis. This includes new reactions and novel synthetic approaches utilizing known catalysts, the discovery or modification of new catalysts, elucidation of catalytic mechanisms through cutting-edge investigations, practical enhancements of existing processes, as well as conceptual advances in the field. Contributions to ACS Catalysis can encompass both experimental and theoretical research focused on catalytic molecules, macromolecules, and materials that exhibit catalytic turnover.