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Dissimilar Reactions and Enzymes for Psilocybin Biosynthesis in Inocybe and Psilocybe Mushrooms

Tim Schäfer, Fabian Haun, Bernhard Rupp, Dirk Hoffmeister

Angewandte Chemie International Edition September 21, 2025 DOI: 10.1002/anie.202512017 (opens in new tab)

Study at a glance

AI-extracted from the abstract
Characteristics In vitro study Peer reviewed
Topics Psilocybin
Citations 7
Key points The psilocybin biosynthetic pathway in Inocybe corydalina is branched and uses different enzymes than in Psilocybe species, indicating independent evolution of psilocybin production.

Abstract

Abstract Psilocybin (4‐phosphoryloxy‐ N , N ‐dimethyltryptamine, 1 ) is the main indolethyl‐amine natural product of psychotropic (so‐called “magic”) mushrooms. The majority of 1 ‐producing species belongs to the eponymous genus Psilocybe , for which the biosynthetic events, beginning from l ‐tryptophan ( 2 ), and the involved enzymes have thoroughly been characterized. Some Inocybe (fiber cap) species, among them Inocybe corydalina , produce 1 as well. In product formation assays, we characterized four recombinantly produced biosynthesis enzymes of this species in vitro: IpsD, a pyridoxal‐5′‐phosphate‐dependent l ‐tryptophan decarboxylase, the kinase IpsK, and two near‐identical methyltransferases, IpsM1 and IpsM2. The fifth enzyme, the insoluble monooxygenase IpsH, was analyzed in silico. Surprisingly, none of the reactions intrinsic to the 1 pathway in Psilocybe species takes place in I. corydalina . Contrasting the situation in Psilocybe , the Inocybe pathway is branched and leads to baeocystin (4‐phosphoryloxy‐ N ‐methyltryptamine, 3 ) as a second end product. Our results demonstrate that mushrooms recruited distantly or entirely unrelated enzymes to evolve the metabolic capacity for 1 biosynthesis twice independently.