(280e) Gram-Scale Production of the Psychedelic Natural Product, Psilocybin, in E. coli

The development of a psilocybin production platform in a highly engineerable microbe could lead to rapid advances towards the bioproduction of psilocybin for use in ongoing clinical trials for treatment resistant depression, post-traumatic stress disorder (PTSD), and addiction. Here, we present our on-going progress towards the development of a microbial cell factory for the production of the hallucinogenic natural product, psilocybin. We will address our efforts in traditional genetic and fermentation conditions optimization, which have yielded titers over 1.0 g/L in fed batch fermentation. This process includes screening of various defined and random transcriptionally varied promoter libraries as well as a combinatorial analysis on the effects of varying induction conditions, temperature, and media supplements on overall product titer.

Ongoing and future efforts to deregulate and enhance intracellular precursor and co-factor flux in our psilocybin production host will also be presented. Specifically, we have targeted the serine, indole, and S-Adenosyl-L-methionine (SAM) biosynthesis pathways and their associated transcriptional regulation. The evaluation of strains and conditions was accomplished through the use of medium-throughput screening in 48-well plates coupled with HPLC analysis of metabolite concentrations. We discuss our work with this unique approach to develop and characterize the production landscape of our novel production host. It can be expected that new and unpublished results will be presented.