ScopeHuman milk is widely acknowledged as the best food for infants, and that is not just because of nutritional features. Human milk also contains a plethora of bioactive molecules, including a large set of human milk oligosaccharides (hMOs). Especially fucosylated hMOs have received attention for their anti-adhesive effects on pathogens by preventing attachment to the intestinal wall. Because hMOs are generally challenging to produce in sufficient quantities to study and ultimately apply in (medical) infant formula, hMO mimics are interesting compounds to produce and evaluate for their biological effects.Methods and resultsWe investigated the digestion, fermentation, and pathogen anti-adhesive capacity of the novel hMO mimic di-fucosyl-β-cyclodextrin (DFβCD). We establish that DFβCD is not digested by α-amylase and also resists fermentation by the microbiota from a 9 month-old infant. In addition, we reveal that DFβCD blocks adhesion of enterotoxigenic E. coli (ETEC) to Caco-2 cells, especially when DFβCD is pre-incubated with ETEC prior to addition to the Caco-2 cells.ConclusionOur results suggests that DFβCD functions through a decoy effect. We expect that our results inspire the generation and biological evaluation of other fucosylated hMOs and mimics, to obtain a comprehensive overview of the anti-adhesive power of fucosylated glycans.