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Bacillus sp. possesses strong fibrinolytic properties. Out of 25 samples collected from different soil and water sources, 25 isolates were selected on the basis of colonial characteristics and Gram staining as Bacillus sp. Out of 25 isolates, 4 were selected based on the result of ?- hemolysis and proteolytic activity. The result of invitro clot lysis and anti-thrombolytic assay revealed that N16 isolates showed highest activity (85%) and strong lysis with 100µl and 150µl of crude enzymes as compared to other 4 isolates. The molecular identification done by 16s r RNA sequencing of the selected 5 isolates showed 100% similarity with N-10 with Bacillus wiedmannii, N-14 with Bacillus safensis, N-15 with Bacillus inaqusorum, N-16 with Bacillus paramycoides. The study helps to find the role of fibrinolyticenzymes produced by Bacillus sp. with highest potential and to overcome the undesirable effects of commercially available thrombolytic agents.