Objective: This study aimed to prepare a kind of mushroom paste with Lentinus edodes, Pleurotus eryngii and Agaricus bisporus as raw materials by enzymolysis with cellulase. Methods: The response surface method was used to optimize the optimal enzymatic hydrolysis process of the mushroom paste with amino acid nitrogen content as the index. Results: The optimal enzymolysis conditions were as follows: enzyme dosage 0.6%, pH 6, enzymolysis temperature 50 ℃, solid-liquid ratio 1∶30 （g/mL）. Under these conditions, the actual amino acid nitrogen content was 0.418 μg/100 g. Conclusion: The sensory evaluation and nutritional indexes of the mushroom paste are up to or better than the national standard, which indicates that the process is feasible and has certain popularization and application values.