The aims of the present study were to optimize the operational parameters to maximize the yield of ultrasound-assisted polysaccharide extraction from Volvariella volvacea (straw mushroom) fruiting bodies by using for the first time one-factor-at-a-time and three-level Box-Behnken factorial designs. A maximum polysaccharide yield of 8.28 ± 0.23% was obtained under the optimized conditions of ultrasound power of 175 W, extraction temperature of 57 °C, extraction time of 33 min, and the ratio of liquid to raw material of 25:1, respectively. Compared to the hot-water extraction, the ultrasound-assistance favored the extraction of polysaccharides from V. volvacea for its higher polysaccharide yield and efficiency. Further preliminary polysaccharide structural characterization indicated that ultrasound treatment affected the monosaccharide compositions and ratios, and molecular weight range of polysaccharides extracted from V. volvacea.

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