The composition and anti-tumor bioactivity of the exopolysaccharide monosaccharide was carried out by adding Rhizoma gastrodiae alcohol extract to the liquid fermentation system of Grifola frondosa. The HPLC was applied to analyse the monosaccharide composition of exopolysaccharide samples and results showed that the crude exopolysaccharide samples were composed of glucose, mannose, and a small amount of galactose. Whereas the mannose, rhamnose, glucose, galactose, arabinose, fucose etc were the main monosaccharide composition of pure exopolysaccharides. Furthermore, the addition of Rhizoma gastrodiae alcohol extract had a significant change in the content of the monosaccharide component of the exopolysaccharide, but did not produce the new monosaccharide component of the polysaccharide. In the sample of pure exopolysaccharides, the monosaccharide molar ratio of mannose:rhamnose:glucose:galactose, arabinose:fucose was 13.8∶3.9∶5∶3.7∶1∶1.7, while the pure exopolysaccharides fermented with Rhizoma gastrodiae monosaccharide mole ratio was 12.7∶3.2∶5.6∶3.5∶1∶1.6. The results about of the biological activity Grifola frondosa exopolysaccharide on mice macrophage (RAW264.7) showed that the pure exopolysaccharides fermented with Rhizoma gastrodiae had obvious activation on macrophage. The pure exopolysaccharides and exopolysaccharides obtained by fermentation with Rhizoma gastrodiae had certain activation on macrophage. The activation of macrophages by crude exopolysaccha-rides was not significant. In vitro, four polysaccharide samples showed no inhibitory effect on human hepatoma HepG 2 cells.