The highest concentrations of psilocybin and baeocystin in six species of higher fungi were obtained by extraction with pure methanol. Psilocin content appeared higher when using aqueous alcohol solutions, but this was an artifact of enzymatic breakdown of psilocybin. Extraction with dilute acetic acid outperformed water-containing alcohols. Methanol extraction remains the safest one-step method for obtaining genuine alkaloids from fungal biomass.
Mycelial cultures of Psilocybe cubensis that naturally produce psilocybin and psilocin can efficiently add a hydroxyl group to tryptamine derivatives at the 4-position. When synthetic N,N-diethyltryptamine was added, the fungus converted it into high amounts of 4-hydroxy-N,N-diethyltryptamine (up to 3.3%) and smaller amounts of 4-phosphoryloxy-N,N-diethyltryptamine (0.01–0.8%) in the fruiting bodies. This marks the first example of directed biosynthesis of tryptamine substances by fungi. Surface cultures of Psilocybe semilanceata also effectively transformed N-methyltryptamine. Baeocystin, a possible natural precursor of psilocybin, was detected and quantified in the biomasses, but no alkaloids were found in the culture medium.