Facchini, Peter JamesDinsmore, Donald Reed2015-12-162015-12-162015-12-162015Dinsmore, D. R. (2015). Functional Genomics and Metabolite Profiling as Tools for Alkaloid Biosynthetic Gene Discovery (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/26246http://hdl.handle.net/11023/2686The benzylisoquinoline alkaloids (BIAs) are diverse group of plant specialized metabolites found in the families Papaveracea, Ranunculaceae, Berberidaceae and Menispermaceae. Plants remain the only commercial source for BIAs and their biosynthesis is poorly understood. O-methyltransferases (OMTs) are wide spread in BIA biosynthesis. Putative OMTs were found in stem and root Next-Generation Sequencing transcriptomic databases. Putative OMT cDNAs were isolated from Papaver somniferum and commercially synthesized. Recombinant protoberberine 2-O-methyltransferase (2OMT) was heterologously expressed in Escherichia coli and assayed. 2OMT demonstrated the 2-O-methylation of protoberberine alkaloids and the 7-O-methylation of simple BIAs. The substrate range and tissue specific expression of 2OMT suggest its in vivo role is converting (S)-cheilanthifoline to (S)-sinactine. A LC-MS based targeted alkaloid profiling of twenty BIA producing species from the families Papaveracea, Ranunculaceae, Berberidaceae and Menispermaceae was conducted.engUniversity of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.BioinformaticsBotanyGeneticsMicrobiologyBiology--MolecularPlant PhysiologyChemistry--AnalyticalBiochemistryPlantsMetabolismMass spectrometryEnzymeOpium poppymaster thesis10.11575/PRISM/26246