Total Biosynthesis of the Tubulin-Binding Alkaloid Colchicine
Total Biosynthesis of the Tubulin-Binding Alkaloid Colchicine
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DOI：
10.1021/jacs.1c08659
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发表时间：
2021-11-24
影响因子：
15
通讯作者：
Sattely, Elizabeth S.
中科院分区：
文献类型：
--
作者：
Nett, Ryan S.;Sattely, Elizabeth S.
秋水仙碱(1)是一种从秋水仙属和金凤花属植物中提取的生物活性植物生物碱，被用作治疗炎性疾病的药物，包括痛风性关节炎和家族性地中海热。这种生物碱的活性归因于其结合微管蛋白二聚体和抑制微管组装的能力，这不仅促进了抗炎作用，而且使秋水仙碱成为一种有效的有丝分裂毒药。秋水仙碱生物合成的生化起源已经研究了50多年，但直到最近才清楚潜在的酶机制。在这里，我们报道了从荣耀草中发现的多种途径酶，这些途径酶允许重组到1的完整代谢途径。这包括三种酶，它们通过剪裁氮原子将先前建立的含tropolone的中间体加工成1。我们进一步证明了在模式植物中通过异源生产从初级代谢物中合成对映纯(-)-1的全生物合成，从而为该药用生物碱的代谢工程提供了未来的努力。此外，我们的研究结果为秋水仙碱生物合成所需的后期修饰的时间和组织特异性提供了见解，这可能与本地生产植物中这类药用生物碱的生物学功能有关。
Colchicine (1) is a bioactive plant alkaloid from Colchicum and Gloriosa species that is used as a pharmaceutical treatment for inflammatory diseases, including gouty arthritis and familial Mediterranean fever. The activity of this alkaloid is attributed to its ability to bind tubulin dimers and inhibit microtubule assembly, which not only promotes anti-inflammatory effects, but also makes colchicine a potent mitotic poison. The biochemical origins of colchicine biosynthesis have been investigated for over 50 years, but only recently has the underlying enzymatic machinery become clear. Here, we report the discovery of multiple pathway enzymes from Gloriosa superba that allows for the reconstitution of a complete metabolic route to 1. This includes three enzymes that process a previously established tropolone-containing intermediate into 1 via tailoring of the nitrogen atom. We further demonstrate the total biosynthesis of enantiopure (-)-1 from primary metabolites via heterologous production in a model plant, thus enabling future efforts for the metabolic engineering of this medicinal alkaloid. Additionally, our results provide insight into the timing and tissue specificity for the late stage modifications required in colchicine biosynthesis, which are likely connected to the biological functions for this class of medicinal alkaloids in native producing plants.