Department of Biology, Concordia University, Montréal, QC, Canada.
Centre for Applied Synthetic Biology, Concordia University, Montréal, QC, Canada.
Nat Commun. 2020 Jul 3;11(1):3337. doi: 10.1038/s41467-020-17172-x.
The tetrahydroisoquinoline (THIQ) moiety is a privileged substructure of many bioactive natural products and semi-synthetic analogs. Plants manufacture more than 3,000 THIQ alkaloids, including the opioids morphine and codeine. While microbial species have been engineered to synthesize a few compounds from the benzylisoquinoline alkaloid (BIA) family of THIQs, low product titers impede industrial viability and limit access to the full chemical space. Here we report a yeast THIQ platform by increasing production of the central BIA intermediate (S)-reticuline to 4.6 g L, a 57,000-fold improvement over our first-generation strain. We show that gains in BIA output coincide with the formation of several substituted THIQs derived from amino acid catabolism. We use these insights to repurpose the Ehrlich pathway and synthesize an array of THIQ structures. This work provides a blueprint for building diverse alkaloid scaffolds and enables the targeted overproduction of thousands of THIQ products, including natural and semi-synthetic opioids.
四氢异喹啉（THIQ）部分是许多生物活性天然产物和半合成类似物的优势结构。植物制造了超过 3000 种 THIQ 生物碱，包括阿片类药物吗啡和可待因。虽然已经对微生物进行了工程改造，以合成几种苯并异喹啉生物碱（BIA）家族的 THIQ 化合物，但低产物滴度阻碍了工业可行性，并限制了对完整化学空间的访问。在这里，我们通过将（S）-网状碱（BIA 的核心中间产物）的产量增加到 4.6 g/L，报告了一个酵母 THIQ 平台，比我们的第一代菌株提高了 57000 倍。我们表明，BIA 产量的提高与氨基酸分解代谢衍生的几种取代的 THIQ 的形成相一致。我们利用这些见解重新利用 Ehrlich 途径并合成了一系列 THIQ 结构。这项工作为构建多样化的生物碱支架提供了蓝图，并能够靶向过量生产数千种 THIQ 产品，包括天然和半合成阿片类药物。
