Mushroom polysaccharides are a type of bioactive macromolecular which isolated from fruiting bodies, mycelia or fermentation broths of edible or medicinal fungus. Recently, mushroom polysaccharides have attracted a lot of attention for regulating gut microbiota via reducing the levels of pathogens and stimulating the growth of beneficial microorganisms, thus creating new possibilities for their use in nutraceutical and functional foods industries. The current review summarizes the isolation, purification and structural characterization methods of mushroom polysaccharides, the degradation of mushroom polysaccharides in intestine, the impacts of mushroom polysaccharides on gut microbiota community and short chain fatty acids (SCFAs) productivity, and the beneficial effects of mushroom polysaccharides to host by targeting gut microbiota. We hope this article can offer some theoretical bases and inspirations for the mechanism study of the bioactivity of mushroom polysaccharides.