Extended Data Fig. 6: Characterization of SoBAHD1.
a. Structure of saponarioside B (13), a product of SoBAHD1 when acting in combination with the S. officinalis enzymes required to produce 12. Modification performed by SoBAHD1 has been highlighted and a table showing relevant calculated adducts and fragments of 13 included. b. N. benthamiana leaves transiently co-expressing various genes were extracted and analysed using HPLC-MS, representative (n = 6) extracted ion chromatograms (EIC) and MS/MS spectra are shown. EIC displayed are for m/z 1699.7206, the calculated mass of the [M-H]- adduct of 13. The negative controls used were extracts from N. benthamiana leaves co-expressing only AstHMGR (tHMGR control) or co-expressing the S. officinalis genes required to produce 12 (tHMGR, SobAS1, CYP716A379, CYP72A984, SoCSL1, UGT73DL1, UGT73CC6, UGT74CD1, SoSDR1, UGT79T1, UGT79L3, UGT73M2 and SoGH1) (QA-TriF(Q)RXX). The additional activity of SoBAHD1 produced two product peaks (13 and 13′) which are identified as SpB and SO1699 (see Extended Data Fig. 7) by comparison to authentic standards.
