Fibrinolytic enzymes from actinomycetes

Fibrinolytic enzymes have nowadays found important place in the medical field. Only few studies had reported the fibrinolytic enzyme production from actinomycetes. So the microbial fibrinolytic enzymes, especially those from actinomycetes, have the potential to be developed as drugs to prevent cardiovascular diseases. Actinomycetes were producing strong enzymes with no side effects, non pathogenic and non expensive. In the present study actinomycetes were isolated from slaughter house soil samples of in and around Salem. Out of 20 actinomycete strains isolated, 7 were found protease positive and further used for the screening for fibrinolytic enzymes on fibrin agar medium. The actinomycete strain S4 was found as potential strain. The actinomycetes species S4 was further subjected for submerged fermentation for 5 days at 28oC and the supernatant of the culture filtrate was processed for characterization of enzyme and fibrinolytic activity. The optimization studies indicated that the best carbon source as sucrose, nitrogen source as yeast extract, pH of 6 and 7, temperature of 28oC and incubation time of 6 days were the favourable parameters for the effective production of fibrinolytic enzyme from actinomycetes stain S4. The purification of the enzyme was done by ammonium sulphate precipitation and dialysis. Protein concentration was determined by the method of Lowry’s using bovine serum albumin as standard and the molecular weight of the enzyme was determined by SDS-PAGE. The protein concentration was measured about 0.47 mg/ml and molecular weight of the enzyme based on electrophoresis was determined as 43 kDa. The in vitro assay for fibrinolytic activity using fibrin plate method revealed that the complete degradation of fibrin within 1 hour. The in vitro degradation of artificial blood clot within 2 hours clearly confirmed the fibrinolytic activity of the enzyme. The in vitro degradation of artificial blood clot within 2 hours confirmed the fibrinolytic activity of the enzyme. Thus the present study gains its novelty by bringing new information to literature on the possibility of exploring actinomycetes as a potential microbial source for the production of an efficacious fibrinolytic enzyme.