Noscapine is an antitumor alkaloid from opium poppy that binds tubulin, arrests metaphase, and induces apoptosis in dividing human cells. Elucidation of the biosynthetic pathway will enable improvement in the commercial production of noscapine and related bioactive molecules. Transcriptomic analysis revealed the exclusive expression of 10 genes encoding five distinct enzyme classes in a high noscapine-producing poppy variety, HN1. Analysis of an F(2) mapping population indicated that these genes are tightly linked in HN1, and bacterial artificial chromosome sequencing confirmed that they exist as a complex gene cluster for plant alkaloids. Virus-induced gene silencing resulted in accumulation of pathway intermediates, allowing gene function to be linked to noscapine synthesis and a novel biosynthetic pathway to be proposed.

Morphinan-based painkillers are derived from opium poppy ( L.). We report a draft of the opium poppy genome, with 2.72 gigabases assembled into 11 chromosomes with contig N50 and scaffold N50 of 1.77 and 204 megabases, respectively. Synteny analysis suggests a whole-genome duplication at ~7.8 million years ago and ancient segmental or whole-genome duplication(s) that occurred before the Papaveraceae-Ranunculaceae divergence 110 million years ago. Syntenic blocks representative of phthalideisoquinoline and morphinan components of a benzylisoquinoline alkaloid cluster of 15 genes provide insight into how this cluster evolved. Paralog analysis identified P450 and oxidoreductase genes that combined to form the gene fusion essential for morphinan biosynthesis in opium poppy. Thus, gene duplication, rearrangement, and fusion events have led to evolution of specialized metabolic products in opium poppy.

Morphinan alkaloids from the opium poppy are used for pain relief. The direction of metabolites to morphinan biosynthesis requires isomerization of (S)- to (R)-reticuline. Characterization of high-reticuline poppy mutants revealed a genetic locus, designated STORR [(S)- to (R)-reticuline] that encodes both cytochrome P450 and oxidoreductase modules, the latter belonging to the aldo-keto reductase family. Metabolite analysis of mutant alleles and heterologous expression demonstrate that the P450 module is responsible for the conversion of (S)-reticuline to 1,2-dehydroreticuline, whereas the oxidoreductase module converts 1,2-dehydroreticuline to (R)-reticuline rather than functioning as a P450 redox partner. Proteomic analysis confirmed that these two modules are contained on a single polypeptide in vivo. This modular assembly implies a selection pressure favoring substrate channeling. The fusion protein STORR may enable microbial-based morphinan production.

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A study of the hydrodynamic and mass transfer performance for Karr reciprocating plate extraction columns has been presented for a range of operating conditions and column diameters of 50, 100, and 450 mm. Although the use of Karr columns has been widespread for many years for a range of important applications, the need for more reliable methods for predicting the performance and scale-up of such columns continues to be a matter of significant importance. The present study has examined the hydrodynamic performance in terms of the dispersed phase hold up and droplet size distribution and mass transfer performance which has incorporated the effects of backmixing in the continuous phase. Work was initially carried out in a 50 mm diameter laboratory scale Karr column using the system 10 v/v% tributyl phosphate/kerosene (continuous)-phenol-water (dispersed) where hydrodynamic and mass transfer performance was analyzed for both directions of mass transfer. Using these data, models were investigated and developed to predict performance over a range of operating conditions. An alternative system consisting of an organic solvent (continuous)-phenolic alkaloid-aqueous caustic (dispersed) was also studied, using both 100 and 450 mm diameter Karr columns. These data were used to validate the hydrodynamic and mass transfer performance models developed for the phenol system in the 50 mm diameter column. Dispersed phase holdup data were found to fit the correlation presented by Kumar and Hartland [Ind. Eng. Chem. Res. 1995, 34, 3925-3940], and the drop size distribution also agreed with the relationship presented by Kumar and Hartland [Ind. Eng. Chem. Res. 1996, 35, 2682-2695 within reasonable accuracy. The mass transfer performance results indicated that the continuous phase controlled the mass transfer rate, and thus, column design was simplified by assuming that the overall mass transfer coefficient can be obtained using a standard mass transfer correlation for the continuous phase. The current mass transfer results were found to be best predicted by a refitted form of the overall mass transfer coefficient correlation developed by Harikrishnan et al. [Chem.

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Highlights Mutants of P450 BM3 can metabolise noscapine. Noscapine is N -demethylated with high selectivity. The metabolites produced are of interest for drug development. The profile of metabolites generated resembles that of mammalian CYP3A4.