Collective Biosynthesis of Plant Spirooxindole Alkaloids through Enzyme Discovery and Engineering

作者：Danni Chu, Hongkui Wang, Zhiwen Nie, Kai Li, Jiaqing Cao, Meijing Yang, Qin Yin, Yang Gu, Yindi Jiang · 发表于：Journal of the American Chemical Society · 年份：2025 · DOI：10.1021/jacs.5c02990 · 被引用次数：14 · 研究领域：Chemical synthesis and alkaloids、Alkaloids: synthesis and pharmacology、Carbohydrate Chemistry and Synthesis

Collective synthesis is a well-established strategy to produce natural product collections from common intermediates. Extending this concept to enzymatic systems, collective biosynthesis offers a sustainable approach to diverse molecular collections but faces distinct challenges, including incomplete pathway knowledge and narrow substrate specificity of biosynthetic enzymes. Here, we overcome these limitations through enzyme discovery and protein engineering to achieve collective biosynthesis of spirooxindole alkaloids, an important class of natural products with diverse biological activities. We identified two key enzymes that catalyze enzymatic epimerization, working sequentially with a previously discovered cytochrome P450 enzyme to fully elucidate the biosynthetic pathway of pentacyclic spirooxindole alkaloids. Structure-guided engineering enhanced substrate recognition capability, enabling collective biosynthesis of 12 tetracyclic and pentacyclic spirooxindole natural products. We also generated new-to-nature fluorinated and deuterated derivatives through precursor-directed biosynthesis. This work provides crucial insights into plant spirooxindole alkaloid biosynthesis while establishing a powerful approach for sustainable production of complex natural products.