This study aimed to compare the in vitro fermentation characteristics of polysaccharides from Bergamot and Laoxianghuang (fermented 1, 3, and 5 years from bergamot) using the stable in vitro human gut fermentation model. Results showed that bergamot polysaccharide (BP) and Laoxianghuang polysaccharides (LPs) with different surface topographies were characterized as mannorhamnan (comprising Mannose and Rhamnose) and polygalacturonic acid (comprising Galacturonic acid and Galactose), respectively. The distinct effects on the gut microbiota and metabolome of BP and LPs may be due to their different monosaccharide compositions and surface morphologies. BP decreased harmful Fusobacterium and promoted beneficial Bifidobacterium, which was positively correlated with health-enhancing metabolites such as acetic acid, propionic acid, and pyridoxamine. Lactobacillus, increased by LPs, was positively correlated with 4-Hydroxybenzaldehyde, acetic acid, and butyric acid. Overall, this study elucidated gut microbiota and the metabolome regulatory discrepancies of BP and LPs, potentially contributing to their development as prebiotics in healthy foods.