Fig. 1: Activity profiles of the A. muciniphila fucosidases on mucin O-glycans.

a Overview of the fucosylated epitopes present in the analyzed conjugated O-glycans from porcine gastric and colonic mucins as well as fetuin. b The fucosidase activity heat map on different epitopes. c An example of enzyme sensitivity toward the fucosylation position in the glycan chain. d–h Examples illustrating the impact of sulphation (red lower case “s”) and sialyl substitutions on fucosidase activity. i–p Examples of activity differences of the α1,2-fucosidases. Enzymes were incubated with mucin substrates blotted on membranes for 24 h, then O-glycans from enzyme-treated and non-treated controls were released and analysed. Relative abundances were calculated by integration of the LC-ESI/MS ion chromatogram area under the curve (AUC) of each glycan peak normalized to the total. Relative abundances are also depicted in the activity heat map in panel b. Data are from a single experiment. The “x” marked data are obtained from a single glycan structure, due to the low abundance of assigned Le^a epitopes based on the MS data. Linkages of the sulphatyl substituent are only given if assignment was feasible. For isobaric (same m/z) glycans the additional number (/n) is the corresponding structure in the LC-ESI/MS data (Supplementary Data File 1). The slightly higher relative abundances compared to the controls reflect the noise due to minor variations in the amount of mucin used in each incubation.