(160aj) Production of Non-Natural, Bioactive, and Potentially Therapeutic Tryptamines Using the Psilocybin Biosynthesis Pathway in an E. coli Host

Natural tryptamines, such as psilocybin and N,N-Dimethyltryptamine (DMT), have structural similarities to the neurotransmitter serotonin and impart vivid hallucinations to humans upon ingestion. Psilocybin, despite being currently listed as a Schedule I drug in the United States, is currently undergoing promising clinical trials to investigate its therapeutic efficacy for a number of indications, including treatment-resistant depression. Our lab has published on the in vivo production of psilocybin in E. coli, which leverages the promiscuity of E. coli’s native tryptophan synthase, TrpB, to synthesize the non-natural tryptophan derivative, 4-hydroxytryptophan, from serine and 4-hydroxindole. 4-hydroxytryptophan is then decarboxylated by the first psilocybin pathway enzyme, PsiD, to form 4-hydroxytryptamine. Here, we investigate whether other non-natural tryptamines could be produced utilizing E. coli’s native tryptophan synthase, along with the genes from the Psilocybe cubensis psilocybin biosynthesis pathway. We tested a panel of 20 substituted indoles in vivo to produce non-natural tryptophan and tryptamine derivatives using our modified E. coli strains. A subset of successfully biosynthesized tryptamine derivatives was then screened against a panel of three N-methyltransferases previously shown to have activity on tryptamines. New compounds with promising yields were then purified and tested for biological activity using a head-twitch response model in rats. This work serves to expand upon the production modalities for non-natural tryptamine derivatives that may have potential for therapeutic use.