Department of Bioengineering, Stanford University, Stanford, CA, USA.
Chan Zuckerberg Biohub, San Francisco, CA, USA.
Nature. 2020 Sep;585(7826):614-619. doi: 10.1038/s41586-020-2650-9. Epub 2020 Sep 2.
Tropane alkaloids from nightshade plants are neurotransmitter inhibitors that are used for treating neuromuscular disorders and are classified as essential medicines by the World Health Organization. Challenges in global supplies have resulted in frequent shortages of these drugs. Further vulnerabilities in supply chains have been revealed by events such as the Australian wildfires and the COVID-19 pandemic. Rapidly deployable production strategies that are robust to environmental and socioeconomic upheaval are needed. Here we engineered baker's yeast to produce the medicinal alkaloids hyoscyamine and scopolamine, starting from simple sugars and amino acids. We combined functional genomics to identify a missing pathway enzyme, protein engineering to enable the functional expression of an acyltransferase via trafficking to the vacuole, heterologous transporters to facilitate intracellular routing, and strain optimization to improve titres. Our integrated system positions more than twenty proteins adapted from yeast, bacteria, plants and animals across six sub-cellular locations to recapitulate the spatial organization of tropane alkaloid biosynthesis in plants. Microbial biosynthesis platforms can facilitate the discovery of tropane alkaloid derivatives as new therapeutic agents for neurological disease and, once scaled, enable robust and agile supply of these essential medicines.
茄科植物中的托烷生物碱是神经递质抑制剂，用于治疗神经肌肉紊乱，被世界卫生组织列为基本药物。全球供应方面的挑战导致这些药物经常短缺。澳大利亚森林大火和 COVID-19 大流行等事件揭示了供应链的进一步脆弱性。需要快速部署的生产策略，这些策略对环境和社会经济动荡具有弹性。在这里，我们从简单的糖和氨基酸开始，通过工程改造酵母来生产药用生物碱莨菪碱和东莨菪碱。我们结合功能基因组学来鉴定缺失的途径酶，通过靶向液泡来实现酰基转移酶的功能表达，利用异源转运蛋白来促进细胞内运输，并进行菌株优化以提高产量。我们的集成系统将来自酵母、细菌、植物和动物的二十多种蛋白质定位于六个亚细胞位置，以重现植物中托烷生物碱生物合成的空间组织。微生物生物合成平台可以促进发现托烷生物碱衍生物作为治疗神经疾病的新治疗剂，并且一旦规模化，就能够为这些基本药物提供强大和灵活的供应。
