Abstract

PRODUCTION, CHARECTERIZATION AND IN-VITRO STUDY OF FIBRINOLYTIC ENZYME FROM LOCALLY ISOLATED MICROCOCCUS LUTEUS B-07

Pallavi K. Aradhye* and Meera D. Chavan

ABSTRACT

Background: Thrombosis, stroke and other cardiovascular diseases are major causes to increase the mortality rate than any other disorders. Microbial fibrinolytic enzyme system is finding very safe, stable and effective treatment on such thrombolytic disorders. Being non-pathogenic, saprophytic in nature Micrococcus luteus B-07 is selected for current study. Objective: To find out the efficiency of microorganism to degrade blood clot at normal physiological conditions. Methodology: Protease producing Micrococcus luteus B-07 was screened on milk nutrient agar plates; its identification was done by biochemical tests and genetic analysis. Production of fibrinolytic enzyme from Micrococcus luteus B-07 was achieved in soy meal wheat powder broth for which conditions were maintained at 330C ±20C and pH 7.2. Fibrinolytic activity checked on fibrin agar plate against plasmin as standard and in-vitro blood clot lyses with 5% of blood clot. The effect of various environmental factors such as temperature, pH and various metal ions on crude enzyme was read at 660 nm. Results: Clear zone of protease production was observed around growth of B-07 isolate. Further in this study it was revealed that the B-07 enzyme produces 665.5 U/ml of fibrinolytic units. Complete dissolution of 5% blood clot within 19 h. The B-07 enzyme showed maximum absorption at 660nm for 40 0C temperature and pH 7.0. The same enzyme gets inhibited with Co++, Cu++, Mg++ and Fe++ ions and the activity was enhanced by Zn++ and Ca++ ions. Conclusion: The fibrinolytic enzyme isolated from Micrococcus luteus B-07 has definite and obvious thrombolytic effects in vitro. The enzyme exhibited similar fibrinolytic activity as that of plasmin on fibrin agar plate. It suggests that the enzyme acts like plasmin without the need of plasminogen activator.

Keywords: Fibrinolytic enzyme, Micrococcus luteus, In-vitro blood clot lysis.