单萜吲哚生物碱 (MIA) 代表一大类植物天然产物，具有针对多种适应症的上市药物活性，包括癌症、疟疾和高血压。卤化 MIA 已显示出改善的药物特性；然而，新型卤代 MIA 的合成仍然是一个挑战。在这里，我们展示了在面包酵母酿酒酵母中从头生物合成两种 MIA（蛇纹石和阿尔斯通宁）的平台，并将其用于系统地探索重构的 MIA 途径用于生产卤化 MIA 的生物催化潜力。由此，我们证明了单个卤代吲哚衍生物可转化为总共 19 种不同的新天然卤代蛇纹石和卤代阿司通类似物。此外，通过微生物 MIA 平台中修饰卤化酶的工艺优化和异源表达，我们记录了氯代阿司通的从头卤化和生物合成。总之，这项研究强调了一个微生物平台，用于酶法探索和生产具有治疗潜力的复杂天然和新天然 MIA。© 2023。作者。

Monoterpenoid indole alkaloids (MIAs) represent a large class of plant natural products with marketed pharmaceutical activities against a wide range of indications, including cancer, malaria and hypertension. Halogenated MIAs have shown improved pharmaceutical properties; however, synthesis of new-to-nature halogenated MIAs remains a challenge. Here we demonstrate a platform for de novo biosynthesis of two MIAs, serpentine and alstonine, in baker's yeast Saccharomyces cerevisiae and deploy it to systematically explore the biocatalytic potential of refactored MIA pathways for the production of halogenated MIAs. From this, we demonstrate conversion of individual haloindole derivatives to a total of 19 different new-to-nature haloserpentine and haloalstonine analogs. Furthermore, by process optimization and heterologous expression of a modified halogenase in the microbial MIA platform, we document de novo halogenation and biosynthesis of chloroalstonine. Together, this study highlights a microbial platform for enzymatic exploration and production of complex natural and new-to-nature MIAs with therapeutic potential.© 2023. The Author(s).