Isolation of lipoxygenase enzyme from patients with allergic rhinitis and study of the effect of natural products isolated from buckthorn plants (rhamnaus L) on its effectiveness

Abstract

This thesis included the isolation and purification of the enzyme lipoxygenase (LOX) from the blood serum of a person with allergic rhinitis disease, the study of several factors affecting its effectiveness, in addition to determining its molecular weight, and then the effect of natural products isolated from buckthorn (rhamnaus L) on the effectiveness of the enzyme partially purified from the blood serum.  In which the DEAD-Cellulose ion exchanger was used for the protein leachate produced by deposition with ammonium sulfate (65-0%) after membrane sorting (dialysis). The package showed high efficacy of the enzyme, as the specific effectiveness was (0.105U/mg) and the number of purification times was (2.77) and the package was relied on in determining the molecular weight of the enzyme by electrical migration technology using SDS-PAGE, which amounted to (66,000) kiloDalton approximately. Then the optimum condition of the effectiveness of the partially purified enzyme were studied and the results showed that the highest effectiveness was at the sixth minute, pH=8, temperature (40°C), (0.6 mmol/L) concentration of substrate (linoleic acid), that the maximum velocity (Vmax) was (184 units / liter), and the value of (Km) by applying Michales – Menton equation and Lineweaver-Burke plot was (0.18) and (0.19) mM respectively. As for the concentration of the enzyme, the results showed that the higher enzyme concentration will increased the activity of enzyme The study also involved isolating natural products from buckthorn (sidr) (flavonoids and oils) by a soxhlet device using various solvents, then the oils were diagnosed with GC gas chromatography and flavonoids were diagnosed with high-performance liquid chromatography technology (HPLC) The results showed that there were several bands in GC chromatography, two of them belong to Eicosenoic acid (Cis11-Eicosenoic acid), while the HPLC was characterized by the presence of five bands belonging to (P-Coumaric acid , Qurectine , Apigenin, Rutin  and Chlorogenic acid).