Discovery of Key Cytochrome P450 Monooxygenase (C20ox) Enables the Complete Synthesis of Tripterifordin and Neotripterifordin

作者：Jiadian Wang, Qin Xie, Xinmeng Wang, Mengfei Long, Yanying Chen, Zheng Liu, Xia Meng, Juan Guo, Zeping Wang, Rongfeng Wang, Siyu Shen, Yun Lü, Yan Yin, Yating Hu, Wei Gao, Xiao Zhang, Ping Su, Luqi Huang · 发表于：ACS Catalysis · 年份：2025 · DOI：10.1021/acscatal.4c07121 · 被引用次数：8 · 研究领域：Bioactive Natural Diterpenoids Research、Phytochemistry and Bioactive Compounds、Plant biochemistry and biosynthesis

C20-oxidized diterpenoids from the ent -kaurane family have long attracted interest because of their intriguing architectures and diverse biological activities. A direct hydroxylation strategy at the inert methyl (20) group of the ent -kaurane framework would simplify their synthesis substantially; however, contemporary chemical access remains a challenge because of their structural complexity. Furthermore, an enzymatic approach is limited by the scarcity of dedicated C20 oxidase reports. Herein, we report a key cytochrome P450 monooxygenase (CYP), C20ox, which catalyzes selective C–H oxidation at C20 of the ent -kaurane scaffold and reveals the complex biosynthetic networks of tripterifordin ( 1 ) and neotripterifordin ( 2 ), two C20-oxidized ent -kaurane diterpenoids with strong anti -HIV activity. We constructed engineered Saccharomyces cerevisiae to produce 1 and 2 from glucose. Simultaneously, we developed a concise chemoenzymatic strategy to synthesize compounds 1 and 2 from steviol. Our findings highlight the effectiveness of this strategy using plant CYPs for the scalable synthesis of C20-oxidized ent -kaurane diterpenoids.