Extended Data Fig. 2: Characterization of GsNMT.
From: Discovery and engineering of colchicine alkaloid biosynthesis
a, Co-expression of GsOMT1 and GsNMT in N. benthamiana with co-infiltrated 1 leads to consumption of putative 2 (m/z 300.1594) and the production of a new compound that corresponds to a methylation (m/z 314.1751), as shown by the LC–MS chromatograms. Activity of full-length GsNMT was confirmed in three separate experiments. b, MS/MS fragmentation spectrum of the generated m/z 314.1751 product (*) at a collision energy of 20 V. This experiment was performed twice, with similar results observed each time. c, Tabulated list and putative structures for ion fragments from MS/MS analysis of the m/z 314.1751 product. See Supplementary Information for a detailed analysis of MS/MS results. d, Untargeted metabolite analysis (XCMS) comparing the presence and absence of GsNMT in the transient co-expression system (n = 6 independent replicates for each experimental condition). The unique mass signatures (P < 0.1 between samples, as determined by XCMS) are shown in ranked order based on their increasing (top) or decreasing (bottom) fold change in abundance between the two conditions. The mass isotopologues (M0 and M1) of the presumed product (m/z 314.1751) are shown in red; the mass isotopologues of the presumed substrate (m/z 300.1594) are shown in blue. e, Proposed reaction catalysed by GsNMT as supported by MS/MS fragmentation and previously published labelling studies.
