A yeast platform for high-level synthesis of tetrahydroisoquinoline alkaloids
A yeast platform for high-level synthesis of tetrahydroisoquinoline alkaloids
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DOI：
10.1038/s41467-020-17172-x
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发表时间：
2020-07-03
影响因子：
16.6
通讯作者：
Martin, Vincent J. J.
中科院分区：
文献类型：
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作者：
Pyne, Michael E.;Kevvai, Kaspar;Martin, Vincent J. J.
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特殊子结构。植物可生产 3,000 多种 THIQ 生物碱，包括阿片类药物吗啡和可待因。虽然微生物物种已被设计用于合成 THIQ 的苄基异喹啉生物碱 (BIA) 家族中的一些化合物，但低产品滴度阻碍了工业可行性并限制了对完整化学空间的利用。在这里，我们报告了一个酵母 THIQ 平台，将中央 BIA 中间体 (S)-网状番荔枝碱的产量提高到 4.6gL(-1)，比我们的第一代菌株提高了 57,000 倍。我们表明，BIA 输出的增加与氨基酸分解代谢产生的几种取代 THIQ 的形成同时发生。我们利用这些见解重新调整艾利希通路的用途并合成一系列 THIQ 结构。这项工作为构建多样化的生物碱支架提供了蓝图，并能够有针对性地过量生产数千种 THIQ 产品，包括天然和半合成阿片类药物。植物合成超过 3000 种四氢异喹啉 (THIQ) 生物碱，但其中只有少数是由工程微生物生产的，且滴度很低。在这里，作者将 (S)-网状番荔枝碱滴度提高至 4.6g/L，并重新利用酵母艾利希途径来合成多种 THIQ 支架。
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 4.6gL(-1), 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. Plants synthesize more than 3000 tetrahydroisoquinoline (THIQ) alkaloids, but only a few of them have been produced by engineered microbes and titers are very low. Here, the authors increase (S)-reticuline titer to 4.6g/L and repurpose the yeast Ehrlich pathway to synthesize a diverse array of THIQ scaffolds.