Isolation, production, purification, assay and characterization of fibrinolytic enzymes (Nattokinase, Streptokinase and Urokinase) from bacterial sources

Nattokinase, Streptokinase and Urokinase are novel fibrinolytic enzymes which are isolated from Bacillus subtilis, β-haemolytic Streptococci and urine sample. The fibrinolytic enzyme Nattokinase, Streptokinase and Urokinase was purified from supernatant of Bacillus subtilis, β-haemolytic Streptococci and recombinant E.coli containing short fragment genomic DNA of Pseudomonas sp. Culture broth and showed thermophilic, hydrophilic, and strong fibrinolytic activity. The optimum temperature and pH of Nattokinase, Streptokinase and Urokinase were 37-55°C and 9, 27-37°C and 7 and 55°C and 9, respectively. The molecular weight of Nattokinase, Streptokinase and Urokinase was approximately 28 kDa, 47 kDa and 34 kDa, respectively, as determined by sodium dodecyl sulfatepolyacrylamide gel electrophoresis. The caseinolytic activity of Nattokinase, Streptokinase and Urokinase were 576.73 U, 467.73 U and 785.73 U, respectively, while fibrinolytic activity achieved by fibrin plate method were 10 U, 5 U and 15 U, respectively.