Angewandte Chemie International Edition
August 1, 2017
Janis Fricke, Felix Blei, Dirk Hoffmeister
210 citations
Psilocybin, the psychoactive compound in magic mushrooms, is synthesized from tryptophan by four fungal enzymes: PsiD (a decarboxylase), PsiK (a kinase), PsiM (a methyltransferase), and PsiH (a monooxygenase). Using a combined reaction of PsiD, PsiK, and PsiM, the authors produced psilocybin enzymatically from 4-hydroxy-L-tryptophan in fewer steps than previously possible. This work identifies the complete biosynthetic pathway, which had been unknown for 60 years, and may enable biotechnological production of psilocybin for pharmaceutical use.
Metabolic Engineering
May 26, 2018
Sandra Hoefgen, Jun Lin, Janis Fricke et al.
113 citations
Expressing multiple genes from a biosynthetic pathway in eukaryotic hosts is challenging because each gene typically requires its own regulatory elements. A new vector system overcomes this by arranging genes as a single polycistron, using a picornavirus-inspired 'stop-carry on' mechanism so that all genes are controlled by one promoter. A split fluorescent reporter gene enables easy selection of transformed colonies. The method successfully produced high yields of the mushroom alkaloid psilocybin by expressing the entire biosynthetic gene cluster in the mould Aspergillus nidulans.
Chemistry - A European Journal
November 14, 2019
Felix Blei, Sebastian Dörner, Janis Fricke et al.
80 citations
The psychotropic effects of Psilocybe 'magic' mushrooms are caused by the alkaloid psilocybin, but their broader secondary metabolome is poorly understood. Analysis of four Psilocybe species identified harmane, harmine, and other tryptophan-derived β-carbolines as natural products, confirmed by NMR spectroscopy and stable-isotope labeling. MALDI-MS imaging showed β-carbolines accumulate toward hyphal apices. As potent monoamine oxidase inhibitors, these β-carbolines are neuroactive and interfere with psilocybin degradation, representing an unprecedented scenario where different natural product pathways from the same building block contribute directly or indirectly to the same pharmacological effects.
Chemistry - A European Journal
July 16, 2018
Janis Fricke, Claudius Lenz, Jonas Wick et al.
52 citations
Psilocybin, the prodrug to the psychotropic compound psilocin, is biosynthesized by numerous mushroom species in the fungal genus Psilocybe and other genera, colloquially known as 'magic mushrooms' for their hallucinogenic effects and recreational use. Clinical trials have recognized psilocybin as a valuable candidate for development into a medication against depression and anxiety. This review highlights the recently elucidated biosynthesis of psilocybin, the concurrently developed concept of enzymatic in vitro and heterologous in vivo production, along with previous synthetic routes. The prospect of psilocybin as a promising therapeutic may entail increased demand, which can be met by biotechnological production. The review also briefly touches on psilocybin's therapeutic relevance and pharmacology.
Chemistry - A European Journal
May 11, 2018
Felix Blei, Florian Baldeweg, Janis Fricke et al.
51 citations
Psilocybin, the main psychoactive alkaloid in 'magic mushrooms,' is being investigated as a potential treatment for depression and anxiety. This work describes an improved method for producing psilocybin enzymatically by adding the mushroom enzyme tryptophan synthase (TrpB) to the reaction. The new route uses cheaper starting materials—4-hydroxyindole and L-serine—to form psilocybin. The same approach also produced two other compounds: a non-natural alkaloid called isonorbaeocystin and the neurotransmitter serotonin. This enzymatic method offers a more cost-effective way to synthesize psilocybin and related molecules for research and potential pharmaceutical use.
Chemistry - A European Journal
February 26, 2020
Janis Fricke, Robert B. Kargbo, Lars Regestein et al.
48 citations
Psilocybin, the main psychoactive alkaloid in Psilocybe mushrooms, is being tested as a treatment for depression. Pharmaceutical psilocybin is currently made by synthetic chemistry. Replacing a difficult chemical phosphorylation step with the mushroom enzyme PsiK allowed production of one gram of psilocybin from psilocin in 20 minutes. A pilot-scale protocol also yielded 150 mg of active, soluble PsiK enzyme. This combination of tryptamine chemistry and enzymatic catalysis may provide access to psilocybin at potentially lower cost.
ChemBioChem
May 18, 2022
Sebastian Dörner, Kai Rogge, Janis Fricke et al.
43 citations
Psilocybe magic mushrooms are best known for producing psilocybin and psilocin, but their broader secondary metabolome is poorly understood. Genomes of five species (P. azurescens, P. cubensis, P. cyanescens, P. mexicana, and P. serbica) revealed much greater and unexplored metabolic diversity than chemical analyses alone. P. cyanescens and P. mexicana were identified as aeruginascin producers. Lumichrome and verpacamide A were also detected as Psilocybe metabolites. These findings support efforts to understand phenomena like paralytic effects attributed to some magic mushrooms.
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.
ChemBioChem
December 4, 2019
Richard Demmler, Janis Fricke, Sebastian Dörner et al.
31 citations
Psilocybe cubensis mushrooms produce psilocybin through a five-step pathway that consumes ATP and SAM, requiring efficient recycling of these co-substrates. The adenosine kinase and SAH hydrolase enzymes from the fungus help regenerate SAM. Gene expression increases in fungal primordia and fruiting bodies. A one-pot reaction combining the methyltransferase PsiM with SAH hydrolase shows that removing the byproduct SAH enables continued biosynthesis.
ChemBioChem
August 11, 2018
Felix Blei, Janis Fricke, Jonas Wick et al.
31 citations
Psilocybe mushrooms produce the psychoactive alkaloid psilocybin from L-tryptophan. A newly identified enzyme, TrpM, mono- and dimethylates L-tryptophan itself, unlike the previously known PsiM enzyme that methylates norbaeocystin. TrpM does not act on tryptamine, indicating a second L-tryptophan-dependent pathway separate from psilocybin biosynthesis. TrpM originated from an ancient duplication of part of the egtDB gene, which codes for an ergothioneine biosynthesis enzyme. This duplicated gene was mostly lost during mushroom evolution but independently re-evolved in various genera. The findings suggest a mechanism where weakly selected genes are preserved by being retained within a widely distributed, conserved metabolic pathway.
Angewandte Chemie
August 1, 2017
Janis Fricke, Felix Blei, Dirk Hoffmeister
26 citations
Psilocybin, the psychoactive tryptamine-derived natural product from Psilocybe mushrooms, has had its biosynthetic enzymes characterized. Four enzymes were identified: PsiD, a new class of fungal L-tryptophan decarboxylase; PsiK, catalyzing the phosphotransfer step; PsiM, a methyltransferase performing repeated N-methylation as the final step; and PsiH, a monooxygenase. A step-economical route synthesized psilocybin from 4-hydroxy-L-tryptophan using PsiD, PsiK, and PsiM in a combined reaction. These findings may enable biotechnological production, given renewed pharmaceutical interest.
Journal of Natural Products
March 5, 2021
Janis Fricke, Alexander M. Sherwood, Adam L. Halberstadt et al.
18 citations
A novel analogue of psilocybin, 5-methylpsilocybin, was produced through a hybrid chemoenzymatic synthesis, combining chemical synthesis of 5-methylpsilocin with enzymatic phosphorylation using a purified kinase from Psilocybe cubensis. The product was isolated with high purity via solvent-antisolvent precipitation. In a mouse head-twitch response assay, 5-methylpsilocybin showed psychedelic-like activity more potent than dimethyltryptamine but less potent than psilocybin.
Microbial Biotechnology
November 1, 2024
Slavica Janevska, Sophie Weiser, Ying Huang et al.
13 citations
By deleting genes involved in L-tryptophan catabolism, psilocybin production was increased fivefold in baker's yeast and tenfold in the filamentous fungus Aspergillus nidulans. Process optimization in A. nidulans batch cultures yielded a final psilocybin titre of 267 mg/L with a space-time-yield of 3.7 mg/L/h. The engineered strain demonstrates suitability as a production chassis for psilocybin and other tryptamine-derived pharmaceuticals.
Fungal biology and biotechnology
April 25, 2024
Paula Sophie Seibold, Sebastian Dörner, Janis Fricke et al.
8 citations
Psilocybin, a psychedelic alkaloid, can account for up to 2% of the dry mass of Psilocybe mushrooms, creating a high demand for its precursor L-tryptophan during carpophore (fruiting body) formation. Using Psilocybe mexicana, researchers found that genes for L-tryptophan biosynthesis (trpE1, trpD, trpB) were upregulated in carpophores, while genes for L-tryptophan-consuming pathways (idoA, iasA) were massively downregulated. The IasA enzyme was characterized as the first microbial L-tryptophan-preferring acetaldehyde synthase. Comparison with Psilocybe cubensis revealed species-specific differences in regulation. This coordination of primary and secondary metabolism routes L-tryptophan toward psilocybin production, providing initial insight into how Basidiomycota manage metabolic flux.
Common Library Network (Der Gemeinsame Bibliotheksverbund)
January 1, 2020
Janis Fricke
The structural diversity and genetic and enzymatic foundations of indole alkaloid biosynthesis in Basidiomycetes of Psilocybe spp. were investigated. Through in silico analyses and heterologous expression, the biosynthesis of psilocybin was elucidated, identifying the responsible genes and biochemically and phylogenetically characterizing the resulting enzymes. Psilocybin biosynthesis is catalyzed by four enzymes: L-tryptophan decarboxylase PsiD, cytochrome P450 monooxygenase PsiH, kinase PsiK, and N-methyltransferase PsiM, which are unique secondary metabolite enzymes whose substrate specificities revealed a sequence differing from the previously postulated pathway.