Plant Biochemistry Laboratory, Department of Plant and Environment Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871, Frederiksberg C, Denmark.
Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100, Copenhagen, Denmark.
Nat Commun. 2022 Aug 25;13(1):5011. doi: 10.1038/s41467-022-32667-5.
The diterpenoid triepoxides triptolide and triptonide from Tripterygium wilfordii (thunder god wine) exhibit unique bioactivities with potential uses in disease treatment and as a non-hormonal male contraceptives. Here, we show that cytochrome P450s (CYPs) from the CYP71BE subfamily catalyze an unprecedented 18(4→3) methyl shift required for biosynthesis of the abeo-abietane core structure present in diterpenoid triepoxides and in several other plant diterpenoids. In combination with two CYPs of the CYP82D subfamily, four CYPs from T. wilfordii are shown to constitute the minimal set of biosynthetic genes that enables triptonide biosynthesis using Nicotiana benthamiana and Saccharomyces cerevisiae as heterologous hosts. In addition, co-expression of a specific T. wilfordii cytochrome b (Twcytb-A) increases triptonide output more than 9-fold in S. cerevisiae and affords isolation and structure elucidation by NMR spectroscopic analyses of 18 diterpenoids, providing insights into the biosynthesis of diterpenoid triepoxides. Our findings pave the way for diterpenoid triepoxide production via fermentation.
雷公藤中的二萜三环氧三萜内酯化合物雷公藤红素和雷公藤乙素具有独特的生物活性，有望用于疾病治疗和作为非激素男性避孕药。在这里，我们表明 CYP71BE 亚家族的细胞色素 P450（CYP）催化了前所未有的 18(4→3)甲基移位，这是二萜三环氧三萜内酯化合物和其他几种植物二萜化合物中存在的 abeo-abietane 核心结构生物合成所必需的。与 CYP82D 亚家族的两个 CYP 结合使用，来自 T. wilfordii 的四个 CYP 被证明是构成最小的生物合成基因集，使使用 Nicotiana benthamiana 和 Saccharomyces cerevisiae 作为异源宿主的雷公藤乙素生物合成成为可能。此外，特定的 T. wilfordii 细胞色素 b（Twcytb-A）的共表达使 S. cerevisiae 中的雷公藤乙素产量增加了 9 倍以上，并通过 NMR 光谱分析分离和阐明了 18 种二萜化合物，为二萜三环氧三萜内酯化合物的生物合成提供了深入了解。我们的发现为通过发酵生产二萜三环氧三萜内酯化合物铺平了道路。
