Psychedelics have shown promise for the treatment of several mental disorders such as post-traumatic stress disorder, anxiety, treatment resistant depression, and addiction. These small biomolecules are difficult to mass produce through chemical means or through extraction from the original organism. Recently, psilocybin has been produced on a gram per liter scale in E. coli using 4-OH indole as a substrate. That psilocybin biosynthetic pathway, consisting of PsiD, PsiK, and PsiM in operonic form, was chromosomally integrated into E. coli using transposase to increase the stability of the genes and the consistency of expression compared to plasmid-based systems. Additionally, the Cas9 and λ-Red recombinase system was used to optimize promoter choice. We present a system of pathway integration and subsequent in vivo optimization as well as the resultant promoter-optimized psilocybin production strain.
Author: Phil O’Dell
Faculty Advisor: J. Andrew Jones, Chemical, Paper, and Biomedical Engineering