Abstract: In this study, extraction of proteins from base of Flammulina velutipes was investigated. Materials were crushed into 100 mesh powders, and 0.14mol/L NaCl buffer was used to extract for twice. Based on single factor experiments, solid-liquid ratio, extraction temperature, extraction time, and extraction pH were optimized using Box-Behnken design and response surface methodology for achieving maximum extraction rate of proteins. Factor contribution rate obtained from the F-test were as follows:solid-liquid ratio >extraction pH >extraction time >extraction temperature. The optimum extraction parameters obtained from response surface analysis were as follows:solid-liquid ratio 1 ∶16g/mL, extraction temperature 30℃, extraction time 30min, and extraction pH10. Under the optimized conditions, the predicated extraction rate of proteins was 64.75%, meanwhile the rate of proteins was 64.91% in the validated experiment with the relative deviation value of 0.25%.