This study was designed to facilitate enzymatic protein hydrolysis of catfish (Clarias batrachus) using subtilisin. The effect of incubation time (30, 60, 120, 180 min) with enzyme concentration of 1% (v/w substrate); pH = 8.0; 55°C was studied to determine the degree of hydrolysis (DH), peptide content, amino acid profile and proximate composition of catfish protein hydrolysate (CPH). Results showed that CPH obtained with maximum incubation time had the highest DH; 59% and peptide content; 7200 μg/ml. Th