Format

Send to

Choose Destination
PLoS One. 2015 Apr 23;10(4):e0124459. doi: 10.1371/journal.pone.0124459. eCollection 2015.

Synthesis of Morphinan Alkaloids in Saccharomyces cerevisiae.

Author information

1
Department of Biology, Concordia University, Montréal, Québec, Canada; Centre for Structural and Functional Genomics, Concordia University, Montréal, Québec, Canada.
2
Centre for Structural and Functional Genomics, Concordia University, Montréal, Québec, Canada.

Abstract

Morphinan alkaloids are the most powerful narcotic analgesics currently used to treat moderate to severe and chronic pain. The feasibility of morphinan synthesis in recombinant Saccharomyces cerevisiae starting from the precursor (R,S)-norlaudanosoline was investigated. Chiral analysis of the reticuline produced by the expression of opium poppy methyltransferases showed strict enantioselectivity for (S)-reticuline starting from (R,S)-norlaudanosoline. In addition, the P. somniferum enzymes salutaridine synthase (PsSAS), salutaridine reductase (PsSAR) and salutaridinol acetyltransferase (PsSAT) were functionally co-expressed in S. cerevisiae and optimization of the pH conditions allowed for productive spontaneous rearrangement of salutaridinol-7-O-acetate and synthesis of thebaine from (R)-reticuline. Finally, we reconstituted a 7-gene pathway for the production of codeine and morphine from (R)-reticuline. Yeast cell feeding assays using (R)-reticuline, salutaridine or codeine as substrates showed that all enzymes were functionally co-expressed 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 S. cerevisiae and pave the way for their complete synthesis in recombinant microbes.

PMID:
25905794
PMCID:
PMC4408053
DOI:
10.1371/journal.pone.0124459
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Public Library of Science Icon for PubMed Central
Loading ...
Support Center