Biosynthesis of salvinorin A proceeds via the deoxyxylulose phosphate pathway.

Salvinorin A, a potent kappa-opioid receptor agonist from the hallucinogenic plant Salvia divinorum, is biosynthesized via the 1-deoxy-D-xylulose-5-phosphate (DXP) pathway, not the classic mevalonic acid pathway. This was determined by feeding microshoots with isotopically labeled precursors and analyzing the resulting salvinorin A (2.7 mg from 200 microshoots) using NMR and HR-ESI-MS. Incorporation of labeled glucose and 1-deoxy-D-xylulose confirmed the DXP pathway, while labeled methionine showed that methylation of the C-4 carboxyl group is catalyzed by a type III S-adenosyl-L-methionine-dependent O-methyltransferase.

Study at a glance

Abstract

Salvinorin A, a neoclerodane diterpenoid, isolated from the Mexican hallucinogenic plant Salvia divinorum, is a potent kappa-opioid receptor agonist. Its biosynthetic route was studied by NMR and HR-ESI-MS analysis of the products of the incorporation of [1-(13)C]-glucose, [Me-(13)C]-methionine, and [1-(13)C;3,4-(2)H2]-1-deoxy-D-xylulose into its structure. While the use of cuttings and direct-stem injection were unsuccessful, incorporation of (13)C into salvinorin A was achieved using in vitro sterile culture of microshoots. NMR spectroscopic analysis of salvinorin A (2.7 mg) isolated from 200 microshoots grown in the presence of [1-(13)C]-glucose established that this pharmacologically important diterpene is biosynthesized via the 1-deoxy-D-xylulose-5-phosphate pathway, instead of the classic mevalonic acid pathway. This was confirmed further in plants grown in the presence of [1-(13)C;3,4-(2)H2]-1-deoxy-D-xylulose. In addition, analysis of salvinorin A produced by plants grown in the presence of [Me-(13)C]-methionine indicates that methylation of the C-4 carboxyl group is catalyzed by a type III S-adenosyl-L-methionine-dependent O-methyltransferase.