Fibrinolytic proteases are enzymes that degrade the fibrin, a main component of the blood clot. Thus, the present study aimed to understand the evaluation of aPTT, TT and PT on plasma treated with a fibrinolytic protease produced by Mucor subtillissimus. To assess the anticoagulant effect, the protease was used in different concentrations of 0.5-2.5mg/mL. The tests showed that the enzyme promoted a significant prolonged time over the PT clotting time as concentration increased. In the aPTT assay, fibrinolytic protease practically did not prolong the clotting time, even with increased enzyme concentration. In TT it was verified that at all times of contact with thrombin, be it 5-30 minutes, there was no interference on the action of thrombin or fibrinogen on the formation of thrombi. Since only the prothrombin time among all those studied was altered, it is suggested that the fibrinolytic protease under study affects the extrinsic pathway of coagulation. Thus, it was possible to observe that the serine protease promoted a significant prolonged time on the clotting time of the PT as the enzyme concentration increased, but the serine protease did not cause any type of change in TT and aPTT.