Uricase Isolated from Seeds of Leek (Allium ampeloprasum), Celery (Apium graveolens) and Arugula (Eruca sativa)


AbstractThis research included isolation of uricase from the seeds of leek (Allium ampeloprasum ), celery(Apium graveolens) and arugula(Eruca sativa). Studied the factors effecting its activity and determination of its molecular weight. One proteinous peak had been isolated by gel filtration (using sephadex )G-100)) from the proteinous supernatant obtained using ammonium sulfate saturation(70%). The apparent molecular weight of the isolated enzymes using gel filtration chromatography were (59600+ 209) Dalton for leek, (59430+ 221) Dalton for celery and (59800+ 190) Dalton for arugula .The results also showed that the optimum conditions for the activity of uricase isolated from seeds of leek, celery and arugula obtained at (100 µg/ml) of enzymes concentration using (100, 100 and 120 mmol/l) of uric acid as a substrate, with borate buffer solution at pH (7.0, 7.0, 6.5 ) for (15) minutes at (40, 40 and 35 C) respectively. Using Line Weaver–Burk plot, the values of maximum velocity (Vmax) and Michaelis constant (Km) were from to be (33.3 µmol/ min) and (83.3 mmol/l) for leek, (40 µmol/ min) and (285.7 mmol/l) for celery while (27 µmol/ min) and (166.6 mmol/l) respectively for arugula. When using 100 mM chemical compounds for mercuric chloride(HgCl2), ferrous chloride (FeCl2) and ethylene diamine tetra acetic acid (EDTA) showed decreased the activity of the enzyme, but calcium chloride (CaCl2), manganese sulphate (MnSO4), magnesium sulphate (MgSO4) and potassium chloride (KCl) increased the activity .