Milk clotting enzymes are one of the most significant cheese making raw materials impacting and regulating milk coagulation activity. Calf rennet is one of the enzymes which extracted from suckling calf abomasum. The rennet extracted was optimized for its clotting time t (s) by varying three levels of temperature (30, 35 and 40 o C), pH (4, 5 and 6) and CaCl 2 concentration (0.1, 0.2 and 0.3 gm/500 ml of milk) using response surface method (RSM) and also its milk clotting activity (MCA) was determined. Based on the optimized result, temperature 39.13 o C, pH 4 and CaCl 2 concentration of 0.21 gm/500ml had resulted minimum clotting time of 91.27 s. Using this minimum clotting time, the strength was found to be 1: 13148, establishing the fact that the calf rennet is the best natural coagulant of milk.