Abstract
A type of illness known as cardiovascular disease affects the heart or blood arteries. The flow of blood to the heart, brain, or body is reduced due to thrombosis (blood clotting). Streptokinase (SK) is an extracellular enzyme that converts plasminogen to plasmin, as a medication in thrombolytic treatment. The current study was focused on streptokinase production and enhanced activity using magnetic nanoparticles. Nutrient agar media and liquid state fermentation at pre-optimized conditions was used. The proteolytic activity of the enzyme was determined by using the casein digestion method and then Biuret test was performed for protein estimation. The product of bacterial culture was extracted, and the partial purification of streptokinase was done by ammonium sulfate precipitation, dialysis and then further chromatographic techniques and gel filtration were applied. Immobilization of streptokinase was done by using Fe3O4 magnetic nanoparticles to enhance its activity and protein yield. The immobilization of SK on magnetic nanoparticles was characterized by UV-Vis spectroscopy and Zetasizer. Fe3O4 magnetic nanoparticle showed absorption peak at 224.4 nm and size of 229.4 nm. When compared to the total activity of gel chromatography (5.10 U/ml), immobilization increased the total activity of SK to 7.106 U/ml. In this way enzyme activity of immobilized streptokinase increased by 1.39-fold. Immobilized enzyme was used in in vivo studies and then compared with native one. The activity analysis of immobilized streptokinase was compared with the native one. Immobilized streptokinase exhibited more thrombolytic activity in rabbits than the native form, as demonstrated by in vivo study.
Keywords: Streptokinase; Immobilized enzyme; Nanoparticle; Chromatography; Streptococcus pyogenes; Thrombosis