Biosynthesis of medicinal tropane alkaloids in yeast

The alkaloid drugs hyoscyamine and scopolamine are synthesized from sugars and amino acids in yeast, using 26 genes from yeast, plants, bacteria and animals, protein engineering and a vacuole transporter to enable functional expression of a key acyltransferase. 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(1,2). Challenges in global supplies have resulted in frequent shortages of these drugs(3,4). Further vulnerabilities in supply chains have been revealed by events such as the Australian wildfires(5)and the COVID-19 pandemic(6). Rapidly deployable production strategies that are robust to environmental and socioeconomic upheaval(7,8)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.