To evaluate the prebiotic potential and intestinal fermentation products of wheat bran-derived arabinoxylooligosaccharides (AXOS) in relation to their structure, 5 preparations with structurally different AXOS were included ( approximately 4% wt:wt) in rat diets that mimicked the average Western human diet composition. Xylooligosaccharides (XOS), fructooligosaccharides (FOS), and inulin were used as references. The observed effects mainly depended on the average degree of polymerization (avDP) of the AXOS preparations. The AXOS and XOS preparations with a low avDP (<or=3) resulted in increased colonic acetate and butyrate production and boosted bifidobacteria concentrations in the cecum, but did not significantly lower the concentrations of branched SCFA, which are considered to be markers of protein fermentation by intestinal microbiota. In contrast, an AXOS preparation with a higher avDP (61) effectively suppressed branched SCFA concentrations and thus tipped the balance away from protein fermentation. However, it neither increased colonic butyrate concentrations nor stimulated cecal bifidobacteria development. Two AXOS preparations with a similar avDP (12 and 15) but different average degrees of arabinose substitution (avDAS) (0.69 and 0.27) affected the measured intestinal characteristics similarly, suggesting that the influence of the avDAS was apparently limited and possibly overshadowed by that of the avDP. Among those tested, an AXOS preparation with an avDP of 5 and an avDAS of 0.27 exhibited the best combination of desirable effects on gut health characteristics. Compared with this optimal AXOS preparation, FOS and inulin resulted in similar bifidogenic effects with increased production of colonic acetate (inulin) but not of butyrate. These new insights into the structure-activity relation of AXOS open up new perspectives for the production and application of AXOS preparations with optimized prebiotic and fermentation properties.