Purification of Urokinase Enzyme from Serum Blood of Patients with Pneumonia

Abstract

Aya Jasim Mohammed, Asmaa Hashim Shaker

Among the most common lung diseases such as Tuberculosis, Chronic Obstructive Pulmonary, Asthma, Lung Cancer, Cystic fibrosis, etc., Pneumonia includes all lung diseases that affect the airway and other parts of the lung. Urokinase (UK) [EC 3.4.21.73] is strong plasminogen activator, also known urokinase type plasminogen activator (uPA), which is serine protein. that activates an inactive proenzyme plasminogen to an active plasmin via cleavage of Arg–Val bond. The active plasmin causes break down the fibrin polymers of blood clots. In this study, the UK enzyme was purified from serum blood of patients with pneumonia and the activity of the purified enzyme were measured measured. The purification step includes the precipitation by Ammonium sulfate salt saturation (60%) followed by isolating the enzyme using chromatography Gel filtration by sephadex G–100 and DEAE–Cellulose ion exchange to produce iso-enzyme. The apparent molecular weight of the isolated UK using SDS–PAGE. The results show that the enzyme activity values in male patients is 191.0 ± 11.8 (I.U.) while in female patients is 158.1± 9.59 (I.U.). The enzyme activity in male control 87.8 ± 5.9 (I.U.) and Female 82.5 ± 4.15 (I.U.). Further, the kinetics of the purified enzyme suggest that the value of the reaction rate (Km) is 0.4 M. The enzymes shown an optimum pH at 8.8, and optimum temperature at 37 Cº.