Production of two enzymes, PsiD and PsiH, involved in the psilocybin biosynthesis pathway, in Trichoderma reesei
Student/in: Lisa Becker, BSc
Termin: 16.06.2026, 14:00 Uhr
Ort: Hörsaal F
1. Prüfer/in: Clara Baldin, Phd
2. Prüfer/in: Zuzanna Alicja Dutkiewicz-Kawula, Phd
Vorsitzende/r: Sabine Podmirseg, Mag. Dr. Priv. Doz.
Interessierte Kolleginnen und Kollegen sind herzlich willkommen!
Abstract
Psilocybin is a naturally occurring psychoactive compound produced by several species of the genus Psilocybe and has gained considerable attention due to its promising therapeutic potential in the treatment of depression, anxiety, and addiction disorders. Despite its medical relevance, the biosynthesis of psilocybin remains incompletely understood. In particular, the enzymes PsiD and PsiH, which catalyse key steps in the biosynthetic pathway, have not yet been successfully expressed in heterologous bacterial or yeast systems, limiting their structural and functional characterization.
The aim of this master’s thesis was to establish the heterologous production of the psilocybin biosynthesis enzymes PsiD and PsiH in the filamentous fungus Trichoderma reesei. Expression cassettes containing the respective genes was integrated into the azg1 locus of T. reesei using a CRISPR/Cas9 mediated transformation approach. Successful transformants were selected via 8-azaguanine resistance and verified by genotyping PCR. Subsequently, recombinant strains were cultivated under various culture conditions to identify optimal expression parameters. Protein production was assessed by SDS-PAGE analysis and Western Blot.