Abstract: In order to expend the uses of Agaricus bisporus waste stipe, water was taken as extraction solvent and the effects of extraction temperature, extraction time, ratio of liquid to solid, extraction times, ratio of ethanol to extract and ethanol precipitation time on the yield of polysaccharides from Agaricus bisporus waste stipe were investigated by the single factor experiments.Technology parameters were optimized by orthogonal test.The results showed that the degree of the factors significantly influencing the yield of polysaccharides was in order of extraction times, extraction time, extraction temperature, ratio of liquid to solid. The optimum extraction conditions were as follows: volume of ethanol to extract for precipitation was 2 times, precipitation time was 2h, extraction temperature 85℃, extraction time 90min, ratio of liquid to solid 15∶1, extraction times 3.The yield of polysaccharides could be up to 5.50% under the optimum extraction conditions.