Afini, A.V.M., Sooraj S. Nath, Smitha, K.V., and Kunhi A.A.M. (2016) Production and partial characterization of fibrinolytic enzyme from a soil isolate Aspergillus carbonarius S-CSR-0007. Int. J. Pharm. Pharm. Sci., 8(12): 142-148. Doi. 10.22159/ijpps.2016v8i12.15069

ABSTRACT

Objective: This work was undertaken with the aim of isolating and screening fungal soil isolates with fibrinolytic activity.

Methods: Soil sample near slaughter house was collected and screened for fibrinolytic activity by using fibrin-agar. Enzyme production was optimized under various parameters like pH, temperature, substrate concentration and purified partially by ammonium sulphate precipitation. The stability of the partially purified enzyme was analyzed under the influence of a wide range of pH, temperature, and substrate concentrations.

Results: Among the seven isolates screened, Aspergillus carbonarius S-CSR-0007 exhibited largest clear zone and was selected for further studies. Among the various substrates tested casein was found to support the highest caseinolytic activity of 816 U/ml and fibrinolytic activity of 510 U/ml. The culture supernatant of A. carbonarius S-CSR-0007 was fractionated by ammonium sulfate precipitation followed by dialysis, and maximum activity was obtained in the fraction with 80% ammonium sulfate, with an enzyme activity of 1200 U/ml using tyrosine as standard. The partially purified fibrinolytic enzyme showed optimal activity at 45 °C and pH 7.0. The enzyme was stable up to a temperature of 50 °C and pH 8.0, and the optimum substrate concentration was 4%.

Conclusion: The crude enzyme showed high blood clot lysis activity, which may be a good candidate in the pharmaceutical industry. However, more studies need to be carried out to establish its clinical use.

Keywords: Ammonium sulphate precipitation, Aspergillus carbonarius S-CSR-0007, Fibrin, Fibrinolytic enzyme