Structure dependent fermentation kinetics of dietary carrot rhamnogalacturonan-I in an in vitro gut model

Plant derived dietary polysaccharides are important for gut health and have the potential to modulate the gut microbial community. Dietary rhamnogalacturonan-I obtained by enzymatic treatment of carrot pomace has shown prebiotic properties. In the present study, fermentability of carrot rhamnogalacturonan-I (cRG-I) by faecal microbiota of four donors was studied in an adapted M-SHIME® intestinal model. Despite its complex structure, cRG-I was degraded rapidly in the proximal colon compartment and fermentation became quicker and more complete during three weeks of repeated supplementation. Tracking the change in the molecular weight distribution pattern of cRG-I during the supplementation showed two main donor-dependent gut microbial fermentation strategies designated as either the general or preferential pathway. In the general fermentation pathway, different cRG-I structures were hydrolysed concomitantly, while in the preferential pathway discrete structures were sequentially fermented in a selective manner. Especially arabinan sidechains were utilized before the RG-I backbone, which correlated with an increase in Bifidobacterium longum absolute abundance over the three weeks period. MALDI-TOF MS confirmed that arabinan-, galactan- and arabinogalactan-sidechains were first to be released and degraded. Donor specific production of all SCFA increased over time with a general trend of higher levels of acetate and propionate than butyrate. Strikingly, although the host's baseline gut microbiota composition led to distinct cRG-I hydrolysis routes, the final RG-I consumption was almost complete for both routes, leading to similar metabolic profiles at the end of the three weeks treatment period.