Abstract:[Objective] To research the effect of enzymatic hydrolysate of Lentinula edodes on immune function and intestinal flora in mice. [Methods] Firstly, the immunosuppression model was produced by cyclophosphamide (CY) modeling method. And then one group was filled with no enzymatic hydrolysate by stomach and another group with enzymatic hydrolysate. Lastly, curative effect, immune index, the total number of intestinal bacteria, E. coli, Lactobacillus and Fungi in mice were determined. [Results] Enzymatic hydrolysate of Lentinula edodes could make immune suppression of mice, such as active degree, feces, fur and weight, return to normal levels. It also could improve the spleen and thymus index (P<0.05), promote phagocytic activity of macrophages (P<0.01). Meanwhile, it could improve the number of Lactobacillus which is the main beneficial bacteria in mice intestine (P<0.01) . And it could restrain the growth of pathogenic bacteria such as E. coli (P<0.01). [Conclusion] The enzymatic hydrolysate of Lentinula edodes could improve the immune function and the intestinal microflora environment. It could adjust intestinal function of mice. The enzymatic hydrolysate of Lentinula edodes had high edible and medicinal value.