The bases or stipes of mushrooms are normally discarded as low-economic value animal feed and compost. There are no known reports on deriving polysaccharides from these mushroom wastes for use as prebiotics. This study showed that the relatively low concentration (0.1% to 0.5%) of polysaccharides from Lentinula edodes stipe, Pleurotus eryngii base, and Flammulina velutipes base can enhance the survival rate of Lactobacillus acidophilus, Lactobacillus casei, and Bifidobacterium longum subsp. longum during cold storage. The polysaccharides had synergistic effects with the peptides and amino acids from a yogurt culture to maintain probiotics above 10(7) CFU/mL during cold storage, and they also had significant protective effects on these probiotics in simulated gastric and bile juice conditions to achieve beneficial effects in the host. These results showed that mushroom wastes, which are cheaper than other sources, could be an important, new, alternative source of prebiotics.

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