Extended Data Fig. 3: Activity and affinity of sulfatases to targeted substrates.
From: A single sulfatase is required to access colonic mucin by a gut bacterium
a, Recombinant enzymes (1 μM) were incubated with 1 mM of substrate in 10 mM MES pH6.5 with 5 mM CaCl2 for 16h at 37 °C. Sulfated disaccharides were generated by adding 1 μM of a characterized α1,3/1,4-fucosidase (BT1625) in the enzymatic reaction. Control reactions without sulfatases were carried in the same conditions. Samples were analysed by mass spectrometry and the intensity of the substrate and reaction products was used for comparison of the relative abundance of these sugars after incubation with the respective enzymes. b, Affinity studies looking at the effect of ligand binding on the melting temperature of 3S and 6S-Gal sulfatases. All reactions were performed in 100 mM BTP, pH 7.0 with 150 mM NaCl. For sample melting temperatures see Supplementary Table 11. c, Activity of 3S-Gal/GalNAc sulfatases (10 μM) against 3S-GalNAc (10 mM). Reactions were performed in 10 mM Hepes, pH 7.0, with 150 mM NaCl and 5 mM CaCl2. The data shown are one representative from the biological replicates conducted (n = 3).
