Hyaluronan (HA) has various biological functions and is used extensively as a dietary supplement. Previous studies have shown that the probiotic effects of polysaccharides are closely associated with their molecular properties. The intestinal microbiota has been demonstrated to degrade HA; however, the regulatory effects of different molecular weights (MW) of HA on gut microbiota and metabolites are unknown. In the present study, we performedin vitrofermentation of human-derived feces for three MWs of HA (HA1, 32.3 kDa; HA2, 411 kDa; and HA3, 1510 kDa) to investigate the differences in the fermentation properties of HA with different MWs. We found that gut microbiota can utilize all HAs and, consequently, produce large amounts of short-chain fatty acids (SCFAs). In addition, we showed that all three HA MWs promoted the growth ofBacteroides,Parabacteroides, andFaecalibacterium,with HA1 being more effective at promoting the growth ofBacteroides. HAs have various regulatory effects on the structure and metabolites of the gut microbiota. Spearman’s correlation analysis revealed that alterations in gut microbiota and their metabolites were significantly correlated with changes in metabolic markers. For instance, HA1 enriched α-eleostearic acid and DL-3-aminoisobutyric acid by regulating the abundance ofBacteroides, and HA3 enriched Thymidin by regulatingFaecalibacterium. Collectively, the fermentation properties of HA vary across MW, and our results provide insights into the potential association between the MW of HA and its fermentation characteristics by the gut microbiota. These findings provide insights into the influence of the gut microbiota and HAs on the health of the host.