CARBONIC ANHYDRASE ENZYME FROM THE Alectoris chukar LIVER PURIFICATION CHARACTERIZATION AND ASSESSMENT OF ENZYME KINETICS AGAINST SOME PESTICIDES AND METAL TOXICITY

Abstract

Carbonic anhydrases (CAs) take major roles in many pathological situations associated with convulsions, carcinoma, glaucoma, weight problems, etc. In former researches; using tissues and erythrocytes of a wide range of organisms like; dogs, swine, sheep, chickens, bees, fish, goats, bovines, bacteria, and humans, CA enzymes have been purified and characterized. With this research, the CA enzyme has purified from Alectoris chukar liver tissue with 1.23 EU x mg(-1) of specific activity, 55.5% purification yield, and 460.7 purification folds. The purity of the purified enzyme has been approved by SDS-PAGE. The molecular weight was estimated to be about 28.4 kDa. As the characterization of CA enzyme's in the Alectoris chukar liver has been done; the optimum ionic strength-50 mM Tris-SO4. the optimum pH-8.0, and the optimum temperature-35 degrees C have been decided. K-M and V-max values were calculated by Lineweaver Burk graph. K-M constants were estimated as 1.035 mM, V-max, values as 0.02 molxmin(-1) for 4-nitrophenyl acetate. Inhibitory consequences of some pesticides (Spinosad and Dimethoate) and metal ions (Al3+, Cu2+, Ba2+, Fe2+, Mn2+, Se2+) have been analyzed on the purified CA enzyme. Inhibition graphics were drawn to find the IC50 values of metals and pesticides displaying inhibition.