EC Number |
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2.3.1.150 | Agrobacterium-mediated transfection of Papaver somniferum with the salutaridinol 7-O-acetyltransferase gene, 21 transgenic opium poppy plants over-expressing SalAT are analysed in three separate glasshouse trials, suppression of SalAT by RNAi, SalAT involved in alkaloid accumulation |
2.3.1.150 | expressed as His-tag fusion protein in Sf9 insect cells |
2.3.1.150 | expressed in Escherichia coli as a His-tagged fusion protein |
2.3.1.150 | real-time quantitative PCR expression analysis |
2.3.1.150 | salutaridine synthase (PsSAS), salutaridine reductase (PsSAR) and salutaridinol acetyltransferase (PsSAT) are functionally coexpressed in Saccharomyces cerevisiae and optimization of the pH conditions allowed for productive spontaneous rearrangement of salutaridinol-7-O-acetate and synthesis of thebaine from (R)-reticuline. A 7-gene pathway for the production of codeine and morphine from (R)-reticuline is reconstituted. Yeast cell feeding assays using (R)-reticuline, salutaridine or codeine as substrates show that all enzymes are functionally coexpressed in yeast and that activity of salutaridine reductase and codeine-O-demethylase likely limit flux to morphine synthesis. The results of this study describe a significant advance for the synthesis of morphinans in Saccharomyces cerevisiae and pave the way for their complete synthesis in recombinant microbes |