The umami amino acid and nucleotide contents extracted from 9 kinds of edible fungi were detected by high performance liquid chromatography and the equivalent umami concentration (EUC) value was calculated. Combined with sensory analysis and electronic tongue technology, the umami intensity of 9 kinds of edible fungi was evaluated. The results showed that the sensory score and the umami taste score of the electronic tongue of velvet antler mushroom were the highest among the 9 kinds of edible fungi and the EUC value was 160.56g/100g. Therefore, the velvet antler mushroom was selected as the raw material for the enzymatic extraction process study, and EUC value was used as the investigation index. Combination enzymes of cellulase and flavour protease were selected, and the extraction process of umami amino acids and nucleotides from velvet antler mushroom was optimized by single factor test and response surface method. The optimized enzymolysis conditions were solid-liquid ratio (g/mL) 1∶44.78, cellulase addition 1607.52U/g, flavor protease addition 3044U/g, enzymolysis time 6.14h, pH value 6.50. Three parallel experiments were carried out under optimal condition to obtain the actual EUC value. The actual EUC value was 406.74g/100g, and the relative error with the predicted value was 6.12%.