Fig. 4
From: UGT74S1 is the key player in controlling secoisolariciresinol diglucoside (SDG) formation in flax
Western blot of UGT74S1, UGT7 4S3 (Lus10006353) and UGT74S4 (Lus10014148) and reaction products observed from enzyme assays using purified native enzymes. a, Western blot of the HisTag-purified UGT74S1, UGT74S3 and UGT74S4 probed using AntiXpressTM antibody. The 37 and 50 kDa protein molecular marker sizes corresponding to the Western C precision plus protein marker mixed with conjugant (BioRad) are pinpointed. b, Reaction products formed by each of UGT74S1, UGT74S3 and UGT74S4 after enzyme reaction with SECO as substrate and UDP-glucose or UDP-galactose as sugar donor. SDG secoisolarciresinol diglucoside, SMG secoisolariciresinol monoglucoside, SMGal secoisolariciresinol monogalactoside. c. UPLC chromatograms (280 nm) showing the reaction products of 50 μg of purified proteins for the three UGTs using SECO as substrate and UDP-glucose as sugar donor. Insets show 10× magnification to visualize trace peaks of SMG in UGT74S3 and UGT74S4 reaction products. Appreciable accumulation of SDG (peak 1) and SMG (peak 2) was only observed for UGT74S1. The unreacted SECO (peak 3) is present in all three chromatograms