ChemBioChem
May 31, 2019
Janis Fricke, Alexander M. Sherwood, Robert B. Kargbo et al.
38 citations
Psilocybin and its precursor baeocystin are indole alkaloids from psychotropic Psilocybe mushrooms, currently under clinical investigation for depression and anxiety. A biocatalytic route was developed to synthesize 6-methylated psilocybin and baeocystin from 4-hydroxy-6-methyl-L-tryptophan, using the Psilocybe cubensis enzymes PsiD and PsiK for decarboxylation and phosphorylation, and PsiM for N-methylation. An in silico structural model of PsiM revealed a well-conserved SAM-binding core with peripheral nonconserved elements that likely determine substrate preferences.
Nature Communications
March 28, 2024
Jesse Hudspeth, Kai Rogge, Sebastian Dörner et al.
24 citations
Psilocybin, the natural hallucinogen in magic mushrooms, is produced in a final biosynthetic step where the enzyme PsiM adds two methyl groups to norbaeocystin. Atomic-resolution crystal structures (0.9 Å) of PsiM at different reaction stages reveal its detailed methylation mechanism. Structural and phylogenetic evidence indicates PsiM evolved from METTL16-family RNA methyltransferases, and its bound substrates mimic RNA. Limitations inherited from its ancestral scaffold prevent efficient psilocybin assembly and block trimethylation to aeruginascin. These insights will aid bioengineering efforts to create psilocybin variants with improved therapeutic properties.
ChemBioChem
November 3, 2022
Tim Schäfer, Kristina Kramer, Sebastiaan Werten et al.
20 citations
The enzyme PsiD from the mushroom Psilocybe cubensis initiates the production of psilocybin, the psychedelic compound in magic mushrooms that is being investigated as a treatment for major depressive disorder. Unlike most similar enzymes, PsiD does not require pyridoxal phosphate (PLP) and instead resembles type II phosphatidylserine decarboxylases. Through biochemical experiments and computer modeling, researchers identified and confirmed a non-standard serine protease triad that enables the enzyme to cleave itself into its active form. This finding clarifies the molecular mechanism behind the first step of psilocybin biosynthesis.
Angewandte Chemie International Edition
September 21, 2025
Tim Schäfer, Fabian Haun, Bernhard Rupp et al.
7 citations
Psilocybin, the main psychoactive compound in magic mushrooms, is also produced by some Inocybe species. Researchers characterized four enzymes from Inocybe corydalina and found that none of the reactions used in Psilocybe species occur in this species. Instead, the Inocybe pathway is branched and produces baeocystin as a second end product. These results show that mushrooms evolved the ability to make psilocybin twice independently, using distantly related or entirely different enzymes.
FEBS Letters
October 24, 2024
Kai Rogge, Tobias Wagner, Dirk Hoffmeister et al.
4 citations
Psilocybin, the hallucinogen from magic mushrooms, is being developed as a treatment for depression and other mental health conditions. Its biosynthesis from the amino acid L-tryptophan requires four sequential steps, the third of which is ATP-dependent phosphorylation of the intermediate 4-hydroxytryptamine, catalyzed by the enzyme PsiK. A crystallographic analysis and structure-based mutagenesis study of PsiK reveals how it recognizes its substrate. These findings will aid future bioengineering to create psilocybin variants with improved therapeutic properties.
ChemBioChem
October 16, 2024
Jesse Hudspeth, Kai Rogge, Tobias Wagner et al.
1 citation
The enzyme PsiM from the mushroom Psilocybe cubensis catalyzes the final step in psilocybin biosynthesis, adding two methyl groups to the substrate norbaeocystin. A single amino acid change, M247N, allowed this enzyme to evolve from ancestral monomethylating RNA methyltransferases into a dimethylating enzyme. Mutating this asparagine back to methionine (N247M) or alanine (N247A) eliminated the ability to perform the second methylation. High-resolution crystal structures and kinetic measurements show that Asn247 provides necessary space in the active site for multiple methylations and stabilizes nearby secondary structures through hydrogen bonds, enabling efficient substrate binding and catalysis.
Angewandte Chemie
September 21, 2025
Tim Schäfer, Fabian Haun, Bernhard Rupp et al.
Psilocybin, the main psychoactive alkaloid in magic mushrooms, is produced by several fungal genera. In Psilocybe species the biosynthetic pathway from L-tryptophan is well characterized. This work examined the pathway in Inocybe corydalina, a distantly related mushroom that also makes psilocybin. Four recombinant enzymes from I. corydalina were characterized in vitro: a decarboxylase (IpsD), a kinase (IpsK), and two methyltransferases (IpsM1 and IpsM2); a fifth monooxygenase (IpsH) was analyzed in silico. None of the reactions matched those in Psilocybe. The Inocybe pathway is branched and also produces baeocystin as a second end product. The findings indicate that psilocybin biosynthesis evolved twice independently using unrelated enzymes.