Isolation and Purification of Lipoxygenase from the Serum of Bronchial Asthma Patient and Studying the Effect of Natural Products of Horse Tail ( Equisetum arvense.L) Plant on it's Activity

Abstract

This study includes the isolation and purification of the Lipoxygenase (LOX) enzyme from the serum of an Asthma patient by using different techniques. The result showed the Ion – Exchange chromatography which occurred in the column containing the ion exchange (DEAE- cellulose) diethylamino ethyl – cellulose of the proteinous precipitateth which is produced after dialysis. Two bands of protein and the first band (A) showed the highest activity of the enzyme with the specific activity (2.68 U/mg) and the number times of the purification was (10.30) once. Then, the optimum conditions of LOX enzyme showed the highest activity which it was at the fifth minute at pH = 8 and at (40°C), substrate concentration was about (0.5 mmol/ L) and the maximum Velocity (Vmax) was (168 unit / L), the value of (Km) by using Mikael’s Menten and Linwifer – Burk plot was (0.13) and (0.14) molar respectively. In addition, the determining of the approximate molecular weight of LOX enzyme (the first protein band A) by electrical migration technology (Electrophoresis) using SDS-PAGE and molecular weight and it was (69000) Dalton. Also the study includes the isolation of th the natural products of the horse tail plant (oils) and identificated by GC (Gas-Liquid chromatography) and the flavonoids isolated and identificated by (HPLC) to study their effect on the activity of the partially purified enzyme, the results showed that oils contain an Undecanoic and Lauric acid, while flavonoids contain Catechine, Qurectine, Ferulic acid and Rutin, and it’s effect on the activity of partially purified enzyme showed that flavonoids reduced high-volume lipoxygenase enzyme and then sabonin followed by oils.