How human neuroblastoma cells make morphine.

Recently, our laboratory demonstrated that human neuroblastoma cells (SH-SY5Y) are capable of synthesizing morphine, the major active metabolite of opium poppy. Now our experiments are further substantiated by extending the biochemical studies to the entire morphine pathway in this human cell line. L-[1,2,3-13C3]- and [ring-2',5',6'-2H3]dopa showed high isotopic enrichment and incorporation in both the isoquinoline and the benzyl moiety of the endogenous morphine.

A high-performance liquid chromatography method for the analysis of intermediates of the deoxyxylulose phosphate pathway.

A sensitive and versatile ion pair radio high-performance liquid chromatography (HPLC) method for the investigation of the deoxyxylulose phosphate (DXP) pathway has been developed, allowing the simultaneous separation of phosphorylated, nonphosphorylated, and nucleotide moieties bearing intermediates. Moreover, this method addresses the problem of separating the isomers isopentenyl diphosphate (IDP) and dimethylallyl diphosphate (DMADP). Because the majority of the intermediates of this isoprenoid pathway lack a chromophore, the combination with an on-line radiodetector provides a highly sensitive tool for their detection.

Early steps of deoxyxylulose phosphate pathway in chromoplasts of higher plants.

1-Deoxy-D-xylulose 5-phosphate and 2C-methyl-D-erythritol 4-phosphate have been shown as intermediates of the deoxyxylulose phosphate pathway used for terpenoid biosynthesis in plants and many microorganisms. In plants this non-mevalonate pathway is located in plastids. In order to investigate the formation of five carbon intermediates, chromoplasts from Capsicum annuum and Narcissus pseudonarcissus were incubated with isotope-labeled 1-deoxy-D-xylulose 5-phosphate or 2C-methyl-D-erythritol 4-phosphate.

Studies on the nonmevalonate pathway of terpene biosynthesis. The role of 2C-methyl-D-erythritol 2,4-cyclodiphosphate in plants.

2C-methyl-D-erythritol 2,4-cyclodiphosphate was recently shown to be formed from 2C-methyl-D-erythritol 4-phosphate by the consecutive action of IspD, IspE, and IspF proteins in the nonmevalonate pathway of terpenoid biosynthesis. To complement previous work with radiolabelled precursors, we have now demonstrated that [U-13C5]2C-methyl-D-erythritol 4-phosphate affords [U-13C5]2C-methyl-D-erythritol 2,4-cyclodiphosphate in isolated chromoplasts of Capsicum annuum and Narcissus pseudonarcissus. Moreover, chromoplasts are shown to efficiently convert 2C-methyl-D-erythritol 4-phosphate as well as 2C-methyl-D-erythritol 2,4-cyclodiphosphate into the carotene precursor phytoene.

Biosynthesis of cannabinoids. Incorporation experiments with (13)C-labeled glucoses.

The biosynthesis of cannabinoids was studied in cut sprouts of Cannabis sativa by incorporation experiments using mixtures of unlabeled glucose and [1-(13)C]glucose or [U-(13)C(6)]glucose. (13)C-labeling patterns of cannabichromenic acid and tetrahydrocannabinolic acid were analyzed by quantitative NMR spectroscopy. (13)C enrichments and coupling patterns show that the C(10)-terpenoid moiety is biosynthesized entirely or predominantly (> 98%) via the recently discovered deoxyxylulose phosphate pathway.