A "Magic Mushroom" Multi-Product Sesquiterpene Synthase.

Area of Science:

• Mycology
• Natural Product Chemistry
• Biochemistry

Background:

• Psilocybe mushrooms are known for psychotropic tryptamines.
• The diversity of other specialized metabolites, particularly terpenoids, in these fungi is largely unexplored.
• Understanding these natural products is crucial for assessing their potential role in modulating psychotropic effects.

Purpose of the Study:

• To investigate the metabolic diversity of specialized metabolites in Psilocybe cubensis, focusing on terpenoids.
• To characterize the function of the sesquiterpene synthase CubA.
• To explore the presence of related enzymes in other Psilocybe species.

Main Methods:

• Heterologous expression of the P. cubensis CubA gene in Escherichia coli.
• In vitro enzymatic assays to characterize CubA activity.
• In vivo product formation assays in Aspergillus niger.
• Gas chromatography-mass spectrometry (GC-MS) for metabolite analysis.
• Chromatographic analysis of mature P. cubensis fruiting bodies.

Main Results:

• CubA was confirmed as a multi-product sesquiterpene synthase.
• Major products identified from CubA activity included cubebol, β-copaene, δ-cadinene, and germacrene D, depending on reaction conditions.
• β-copaene and δ-cadinene were also detected in P. cubensis fruiting bodies.
• Genomic analysis revealed enzymes related to CubA in other Psilocybe species.

Conclusions:

• The study elucidates the terpenoid biosynthetic capacity of P. cubensis.
• CubA contributes to the production of diverse sesquiterpenes.
• These findings offer insights into the broader metabolic potential of the Psilocybe genus.