Salvia divinorum (diviner's sage) produces clerodane-type diterpenoids, including the bioactive salvinorin A, the first non-nitrogenous natural compound that acts as an opioid-receptor agonist. Two diterpene synthases were discovered and characterized: SdCPS1, an ent-copalyl diphosphate synthase, and SdCPS2, a clerodienyl diphosphate synthase. SdCPS2 catalyzes the committed step in salvinorin A biosynthesis, supported by its trichome-specific expression and the absence of other class II diTPSs. Structure-guided mutagenesis identified four catalytic residues that allowed reprogramming SdCPS2 to produce four distinct products, advancing understanding of neo-functionalization in plant diterpene synthases and offering potential for synthetic biology platforms.
Peyote (Lophophora williamsii) produces mescaline, a phenethylamine protoalkaloid with psychedelic effects used in Indigenous ceremonies for over 5800 years and now being tested for treating mental disorders. Using transcriptomics and homology-guided gene discovery, researchers identified most enzymes in the mescaline biosynthesis pathway from L-tyrosine, including LwCYP76AD94 (3-hydroxylation), LwTyDC1 (decarboxylation), LwOMT10 and LwOMT2 (O-methylations), and LwNMT (N-methylation). The enzyme catalyzing 5-hydroxylation remains unknown. The findings enable industrial-scale mescaline production via synthetic biosystems, offering an alternative to harvesting the endangered peyote cactus.