Abstract:
The thrombolytic and fibrinolytic enzyme streptokinase is derived from a strain of ß-hemolytic Streptococci. By breaking the fibrin clot, enzyme is commonly utilized in clinical settings to treat cerebral stroke, ischemia, myocardial infarction, and deep vein thrombosis. It works by transforming plasminogen from its zymogen form into active plasmin. Streptokinase production from Streptococcus pyogenes was the main goal of this study. Optimal conditions were used, including a pH of 7.0, a temperature of 38°C, with standard media, and inoculum with a 24-hour incubation period. Ammonium sulfate, ion chromatography, and gel filtration chromatography were used for purification. Enzyme activity was measured at different pH levels, temperatures, and by the application of various inhibitors and activators. Lineweaver-Burk plot was used to calculate the Michaelis-Menten constants, Km and Vmax. Enzyme activation energy was determined and enzyme frequency was calculated by using the Arrhenius equation. With a high recovery rate and fold purification, the enzyme had activity of 9.593 U/ml, protein content of 2.09 mg/ml, and specific activity of 4.589 U/mg. SDS-PAGE analysis was used to quantify the weight of the sample. At different temperatures, enzyme's enthalpy, entropy and free energy were computed. The streptokinase was thermally stable as shown by a negative value for ?S*. Enzyme entropy's negative score indicated a slight degree of instability. Because the free energy for thermal denaturation is larger at higher temperatures, streptokinase is resistant to thermal unfolding. ANOVA was used to see the significance of results at various steps and analysis of means and standard error was done. The results suggested that streptokinase can be used as a potential source to break the fibrin clot, thus can be used for clinical purposes.
Page(s):
71-71
DOI:
DOI not available
Published:
Journal: Abstract Book on Global Science Technology and Management Conference, Volume: 0, Issue: 0, Year: 2023
Keywords:
Streptokinase
,
purification
,
characterization
,
Enzyme production
,
Clinical Use
,
Streptococcus pyogenes