In this study protein hydrolysate was produced from the crucian carp using Alcalase 2.4L.The effect of temperature (40, 45, 50 and 55oC), time (60, 90, 120, 150, 180 & 210min) and enzyme/substrate (Protein) at ratio of (30, 60 and 90 Anson unit), on degree of hydrolysis and antioxidant activity of product were investigated in a in a completely randomized design. The highest degree of hydrolysis was observed at 45oC, after 180min and enzyme/substrate ratio of 60 Anson unit/ Kg substrate. Under these conditions, degree of hydrolysis was 39.38 %. The antioxidant activity of protein hydrolysate was studied using DPPH radical scavenging activity, reducing power and Fe++chelating activity. The most DPPH radical scavenging activity was 57.5%, that was obtained at 45ºC, enzyme activity of 60 Au/kg and 150min of time of hydrolysis. Highest Fe++chelating activity (44.56%) observed at 45oC, enzyme activity of 60 Au/kg, and hydrolysis time of 180min. The highest reducing ability of protein hydrolysate observed at 120min. Hydrolysis time which showed 67.32% reducing power compared to 100 ppm ascorbic acid (100 %).