Purification and Characterization of Fibrinolytic Enzyme Produced by Bacillus subtilis Egy.

Document Type : Original Article

Authors

1 Microbial Chemistry Department, Genetic Engineering and Biotechnology Research Division, National Research Centre, Egypt

2 Faculty of Science, Ain Shams University, Cairo, Egypt

3 Microbial Biotechnology Department, Genetic Engineering and Biotechnology Research Division, National Research Centre, Egypt

Abstract

A novel extracellular enzyme with strong fibrinolytic activity, produced by Bacillus subtilis Egy, which was isolated from the Egyptian soil was purified and characterized. The enzyme was secreted by cultured B. subtilis Egy under solid state fermentation and purified 21.5 purification fold using precipitation by ammonium sulphate followed by ion exchange chromatography, and gel filtration on Sephadex G- 100 chromatography. Purified enzyme showed optimum activity at 50oCreaction temperature and pH 8. Enzyme stability studies revealed that the enzymeis stable up to 30 °C and it retained 80% of its activity at 40oC.In addition, the purified enzyme is stable at pH 8-9. The fibrinolytic activity was enhanced by Mn2+,Cu2+, Ca2+, Na+,  and Ba2+ and inhibited by, Zn2+, and Hg2+. Moreover, the activity was decreased by EDTA and PMSF at 2 mM final  concentration.

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