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Abstract: In the present study, Grifola frondosa strain BUAGF-02 was used for evaluation of glucose and peptone concentrations on extracellular polysaccharides production. Soluble starch, glucose, yeast extract, peptone, etc were used as basic fermentation medium. 5, 10, 15, 20 and 25 g/L of glucose and 1, 1.5, 2, 2.5 and 3 g/L of peptone were used in glucose and peptone assay respectively. The media were shaken and cultured at 25℃ and 170 r/min for 10 d followed by a centrifugation. Mycelia and supernatant were collected. Polysaccharides were collected using the ethanol precipitation method. Both mycelia and polysaccharides were dried and weighted. The results showed that yield of polysaccharides reached the highest level of 0.3 g/100 mL at glucose concentration of 20 g/L, while the highest mycelial weight of 1.82 g/100 mL was achieved at glucose concentration of 25 g/L. The yield of polysaccharides reached the highest level of 0.2 g/100 mL at peptone concentration of 3 g/L, while the highest mycelial weight of 1.26 g/100 mL was obtained at peptone concentration of 2.5 g/L. The study is of certain values to further fermentation researches and applications of G. frondosa polysaccharides.