Blood clots are formed from fibrinogen by thrombin and are lysed by plasmin, which is activated from t issue plasminogen activator. It was developed for the tre atment of thrombosis because of its efficacy and st rong affinity to fibrin. Due to the expense and undesirable side eff ects such as gastrointestinal bleeding, allergic re action, and resistance to repercussion. Therefore the search fo r thrombolytic agents from other source is needed. Production of fibrinolytic enzyme from Bacillus subtilis GBRC1is done by using nutrient broth medium. In addition, a strong fibrinolytic enzyme was purified from the cultivati on media. The purified enzyme was almost homogeneou s, as examined by SDS−PAGE and sephadex G-75 column chrom atography. The enzyme had an optimal pH of 7-12 , a n optimal temperature of 50 °C, for fibrin hydrolysi s. The molecular mass estimated by gel filtration w as 24.6 to 33.0kDa. Phenyl methyl sulphonyl fluoride almost co mpletely inhibited the activity of the enzyme. Futh er studies are necessary for the elucidation of their medicinal ap plications and molecular biological characteristics .