Abstract: Objective:To optimize the fermentation conditions for high-yielding polysaccharide and analyze the polysaccharide composition from mycelia of Pholiota adipose. Methods:The effects of carbon source,nitrogen source and growth factor on the mycelia and polysaccharide content of Pholiota adipose were studied by single-factor experiments,and the medium formulation was optimized by orthogonal test. The effects of temperature,inoculation volume,pH,fermentation time of mycelia were further studied,the fermentation conditions were optimized by orthogonal test,the constituents of the polysaccharides were analyzed by chromatographic. Results showed that,the optimal conditions fermentation of high-yielding Pholiota adiposa mycelia were glucose 2.5%,peptone1%,VB₂ 0.006%,MgSO₄ 0.1%,temperature 25℃,inoculation amount 15%,pH4,fermentation time 10 d,under these conditions,the yield of mycelia was(11.50±0.29) g/L,and the yield of polysaccharide was(1.330±0.035) g/L. In addition,the mycelia of the Pholiota adiposa was composed of L-rhamnose,L-arabinose,and D-galactose. Conclusion:This condition could improve the yield of mycelia polysaccharide and could be used as the fermentation conditions of high-yielding polysaccharide from mycelia of Pholiota adipose.