This work was developed to evaluate the impact of the addition of proteases on the performance characteristics, egg quality, relative weight of digestive organs, and intestinal morphometry of laying hens. 390 Hy Line W36® hens were allocated into five treatments and six replicates with 13 animals. The treatments were: 1) Control (standard formulation), 2) Negative control A -NCA (nutritional reduction according to protease A matrix), 3) Negative control B -NCB (nutritional reduction according to protease B matrix), 4) NCA+protease A (0.250 g/kg of feed) and 5) NCB+protease B (0.125 g/kg of feed). Hens fed the NCA, NCB, and NCA+protease A diets showed reductions in feed intake and egg mass. The addition of protease B provided better results for egg production in both percentage and per dozen as compared to the group fed with the NCA+protease A diets. The hens subjected to diets NCA and NCB showed eggs with a reduced eggshell and thickness percentage. However, supplementation with proteases A and B improved these parameters to values similar to the controls. There was no significant effect of the treatments on the relative weight of the liver, proventricle, gizzard, pancreas, and small intestine. However, the addition of protease A resulted in a decreased value for the relative weight of the large intestine. The jejunum and ileum crypt depths were, respectively, smaller in hens fed the control diet in relation to the NCB diet and the NCA and NCB diets. As it can be concluded, Protease B supplementation provided the best performance results.