Abstract: The methods of enzymatic hydrolysis, the three enzymes hydrolysis together were used to break the cell wall of flammulina velutipes.The contents of proteins, polysaccharides, amino nitrogen and flavor of the extract were examined through single factor test and orthogonal test.This indicated that the most optimal extracting process was adding 2% of three enzymes to hydrolysis for 3h at 40℃, pH6.0. Under this condition, the flammulina velutipes extract contained protein 153.80mg /g, amino nitrogen 13.44mg /g, polysaccharide 49.67mg /g and obteined flavor score 70 points.