2022 Annual Meeting
Towards the In Vivo Production of 5-Methoxy-N,N-Dimethyltryptamine in E. coli
For years, researchers have been interested in psychedelic compounds that have proven to be effective in treating the symptoms of addiction, depression, anxiety, and post-traumatic stress disorder. Because of the prevalence of depression in our world today, we are in desperate need to find treatments that are fast-acting, effective, safe, and relatively inexpensive to produce. For these reasons, I have concentrated my research on the Escherichia coli-based in vivo production of a tryptamine drug with therapeutic potential, 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT). Here, I present the development of a biosynthetic production platform in the highly engineerable microbe, E. coli. A combinatorial gene pathway for 5-MeO-DMT was established by combining PsiD, a tryptophan decarboxylase natively found in psilocybin mushrooms, an O-methyltransferase, and an N-methyltransferase. These genes were expressed in E. coli and induced in the presence of 5-hydroxyindole. The 5-hydroxyindole is condensed with serine through the action of the native tryptophan synthase, TrpB, forming the non-natural amino acid, 5-hydroxytryptophan. Then the activities of the 3 exogenous enzymes were tested, first individually, then together, to assemble a functional pathway for 5-MeO-DMT biosynthesis. Genetic and fermentation conditions such as pH, temperature, induction time, media composition, and promoter strength were varied through controlled experimentation to improve final titers. This production platform provides an alternative method for producing 5-MeO-DMT and is a step towards enabling the bioproduction of psychoactive molecules for both nonclinical and clinical use.