Comprehensive analysis of 42 psilocybin-producing fungal strains reveals metabolite diversity and species-specific clusters.
Jonathan Cohen, Liron Sulimani, Shiri Procaccia, Yaniv Lerenthal, Looz Milay, Ido Taran, Anna Shapira, David Meiri
Scientific reports April 22, 2025 DOI: 10.1038/s41598-025-97710-z via PubMed
Summary
AI-generated from the abstractPsilocybin-producing fungi contain many less-studied metabolites beyond psilocybin, but most research treats them as a single group. By optimizing extraction and analysis methods, the metabolomes of 42 distinct fungal strains across 9 species were examined, revealing broad diversity within and between species. Optimal extraction of fruiting bodies used a 1:20 tissue-to-solvent ratio with 25:75 water:methanol at pH 9 for 1.5 hours, enabling quantification of 8 tryptophan-derived indolamines by HPLC-DAD and identification of putative hydroxypsilocybin by HPLC-MS/MS. A method mimicking in vivo dephosphorylation for in vitro setups was also developed. The work provides a standardized approach for studying these fungi and highlights their pharmaceutical potential.
Study at a glance
| Characteristics | Method development and metabolomic profiling Peer reviewed |
|---|---|
| Sample size | 42 |
| Population | Fungal strains of psilocybin-producing species |
| Topics | Psilocybin |
| Keywords | Dephosphorylation Fungal extraction Fungal metabolome Liquid chromatography Psilocybin-producing fungi |
| Citations | 4 |
| Key finding | Optimized extraction and analysis revealed substantial metabolite diversity across 42 strains from 9 psilocybin-producing species, including quantification of 8 indolamines and identification of putative hydroxypsilocybin. |
Abstract
Psilocybin-producing fungi have garnered attention due to accumulating evidence regarding the therapeutic potential of their principal component psilocybin. This diverse group of fungi harbors a wealth of less-studied metabolites, however, thus far most research has addressed them as a cohesive group. By optimizing an approach for extraction and analysis, we examined the metabolomes of 42 distinct fungi strains and show that the breadth and diversity of metabolites within and between 9 species. We integrated and validated the reproducible and reliable extraction of fruiting bodies followed by chromatographic separation, quantification and identification of their known and yet to be identified secondary metabolites. The optimal extraction of fruiting bodies for high yield of indole alkaloids was achieved using a 1:20 tissue:solvent ratio, 25:75 H2O:MeOH (pH = 9), for 1.5 h, followed by the quantification of 8 tryptophan-derived indolamines by HPLC-DAD and the identification of putative metabolite hydroxypsilocybin by HPLC-MS/MS. The metabolomic analysis revealed the diversity of metabolites within and between species. Finally, we developed and present a method that mimics the in vivo process of dephosphorylation that occurs upon ingestion for in vitro setups. Overall, our study summarizes a standardized approach for both in vitro and in vivo studies involving psilocybin-producing fungi, showcasing the unique metabolome of each strain and the rich diversity of these fungi, encompassing promising pharmaceutical potential.