The effect of graded supplemental doses of an exogenous protease on the growth performance, nutrient digestibility, and nitrogen excretion of white leg shrimp (Litopenaeus vannamei) was assessed: A basal diet formulation mimicking a commercial shrimp feed was supplemented with 0, 100, 200 and 300 mg/kg equivalent to 30,000, 60,000 and 90,000 NFP/kg (diets CTRL, PRO100, PRO200 and PRO300, respectively) of an exogenous protease. Quintuplicate groups of 65 shrimp (IBW = 0.97 ± 0.1 g) were fed one of the four diets. After 62 days of feeding, final body weight (FBW) and specific growth rate (SGR) were not significantly affected by dietary treatments. Shrimp fed PRO200 and PRO300 diets showed a significantly lower feed intake (FI) and feed conversion ratio (FCR) than those fed the CTRL and PRO100 diets. Inversely, shrimp fed the diets with the highest enzyme inclusion (PRO200 and PRO300) demonstrated a significantly higher protein efficacy ratio (PER) than those fed the CTRL and PRO100 diets. Shrimp fed the PRO200 diet benefited from a significantly higher whole-body protein retention than those fed the CTRL diet. Irrespective of the dose, protease supplementation led to a significant increase of dry matter and protein digestibility. At the exception of Isoleucine (Ile) and Leucine (Leu), protease supplementation significantly increased the digestibility of all other amino acids. Shrimp fed all the protease-supplemented diets showed significantly lower excretion of total ammonia nitrogen (TAN), than those fed the CTRL diet. Overall data supports a beneficial effect of supplemental protease in shrimp, supported by an increase of protein and amino acid digestibility, which resulted in a significant improvement of feed utilization criteria (FCR and PER) and a lower excretion of TAN into the water. To demonstrate full efficacy, future trials should target more stringent scenarios of dietary protein supply, including diets formulated with lower crude protein levels and/or protein sources of lower quality.