Extended Data Fig. 9: Functional characterization of Snv11H.

a. Reaction catalyzed by Snv11H and SnvOMT. b. Transient expression of Snv10H, SnvOMT, and Snv11H in N. benthamiana with co-infiltration of strychnine 10. Extracted ion chromatograms for β-colubrine 13 (m/z [M+H]⁺ = 365.1859 ± 0.05, left), 11-deMe brucine 14 (m/z [M+H]⁺ = 381.1808 ± 0.05, middle), and brucine 15 (m/z [M+H]⁺ = 395.1965 ± 0.05, right). This experiment was repeated three times with similar results. c. Transient expression of Snv11H in N. benthamiana with co-infiltration of β-colubrine 13. Extracted ion chromatograms for 11-deMe brucine 14 (m/z [M+H]⁺ = 381.1808 ± 0.05). This experiment was repeated three times with similar results. d. Transient expression of Snv11H and SnvOMT in N. benthamiana with co-infiltration of β-colubrine 13. Extracted ion chromatograms for 11-deMe brucine 14 (m/z [M+H]⁺ = 381.1808 ± 0.05, left) and brucine 15 (m/z [M+H]⁺ = 395.1965 ± 0.05, right). This experiment was repeated three times with similar results. e. Transient expression of SnvOMT in N. benthamiana with co-infiltration of 11-deMe brucine 14. Extracted ion chromatograms for brucine 15 (m/z [M+H]⁺ = 395.1965 ± 0.05). This experiment was repeated three times with similar results. f. In vitro assays using purified SnvOMT from SoluBL21 E. coli. Extracted ion chromatograms for 11-deMe brucine 14 (m/z [M+H]⁺ = 381.1808 ± 0.05, left) and brucine 15 (m/z [M+H]⁺ = 395.1965 ± 0.05, right). This experiment was repeated three times with similar results. g. MS/MS (20 to 50 eV) spectra of generated 11-deMe brucine 14 and brucine 15 (blue) compared to standards (red).