Abstract: In order to determine the hydrolysis degree of different proteases to the protein of Grifola frondosa and the antioxidant activity of the hydrolysates, the pepsin, trypsin, pepsin+trypsin, alkaline protease and papain were used to hydrolyze the protein of Grifola frondosa respectively, and the antioxidant activity of different proteases to the hydrolysates of the protein of Grifola frondosa was determined. The results showed that the hydrolysis degree of the five proteases to the protein of Grifola frondosa was from high to low in the order of alkaline protease>pepsin+trypsin>pepsin>trypsin>papain, and the hydrolysis degree of alkaline protease was the highest, reaching 35.48%. The antioxidant activity of the hydrolysates of alkaline protease, pepsin and pepsin+trypsin increased with the increase of concentration, and the antioxidant activity of the hydrolysates of alkaline protease was the strongest among the three proteases at the same concentration, when the concentration was 0.5 mg·mL^-1 and 1.0 mg·mL^-1, the clearance rate of DPPH free radical was more than 90%, which was similar to that of the control Vc.