Production, Characterization and Evaluation of thrombolytic activity of Staphylokinase of Staphylococcus hominis

Order of Publishing in Issue: 
7
Volume :11
Issue :4
October, 2017 - December, 2017
Page No: 
389-395
Authors: 
C P Chandrappa[1]*, Megha Singh[1], N Chandrasekar[2] and M Govindappa[3]
Address: 
[1]Department of Biotechnology, Shridevi Institute of Engineering and Technology, Sira Road, Tumkur- 572106, Karnataka, India
Address: 
[2]Department of Chemistry, Shridevi Institute of Engineering and Technology, Sira Road, Tumkur-572106, Karnataka, India
Address: 
[3]Department of Biotechnology, Dayananda Sagar College of Engineering, Bengaluru, India

Staphylokinase isolated from Staphylococcus hominis has therapeutic function to dissolve the blood clots. Staphylococcus hominis was isolated from curd. Staphylococcus spp was confirmed by morphological, biochemical and molecular techniques such as 16s rDNA sequencing. Satoh’s medium was used for the production Staphylokinase. Cells were separated from culture broth by centrifugation and the supernatant fluid was added to 3 volume of acetone. After centrifugation of the mixture, the resultant precipitate was purified by ion exchange column chromatography (DEAE Cellulose). Sodium Dodecyl Sulfate Polyacrylamide gel electrophoresis (SDS-PAGE) was done by using 10 to 20% gradient polyacrylamide gel and a 4% stacking gel at 4°C. The purity of the Staphylokinase was determined by SDS-PAGE and HPLC. Finally the thrombolytic activity of Staphylokinase was evaluated by Radial Caseinolytic assay and Heated Plasma agar assay. It was suggested that 0.18 of enzyme is sufficient to dissolve the blood clot.

Keywords: 
Staphylokinase, Staphylococcus hominis, SDS-PAGE, Caseinolytic, Blood clot.
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