Influence of in vitro pectin fermentation on the human fecal microbiome and O-glycosylation of HT29-MTX cells

Pectin is a structurally complex heteropolysaccharide that affects intestinal microorganisms and mucin O-glycans. The present study employed an in vitro model to investigate dynamic changes in microbiota during pectin fermentation. Residual pectin fragments arising from its fermentation were applied to HT29-MTX cells to study the effect of pectin structure on mucin O-glycosylation. Prevotella, Bacteroides, and Parabacteroides were found to preferentially degrade galactose, arabinose, and on the rhamnogalacturonan RG-I side chain region and methyl esterification groups of pectin. Bifidobacterium, Enterococcus, Megamonas, and Dorea metabolized the galacturonic HG region on pectin to produce butyrate. All pectin fragments were found to up-regulate total O-glycans (1.55-2.73 fold) and neutral O-glycans (1.11-1.49 fold) on HT29-MTX mucins. The large HG fragment (81.04 kDa) increased significantly the amount of non-fucosylated glycans (by 2.46-fold); whereas the small HG fragment (16.02 kDa) promoted fucosylated (by 9.25 fold), and especially di-fucosylated O-glycans. Collectively, these results demonstrate that gut microorganisms degrade pectin fragments in the following order of utilization: RG-I, RG-II, and HG. The small fragment of HG improves the expression of fucosylated O-glycans in HT29-MTX cells, mainly owing to an increase in di-fucosylated O-glycans.