Purification of catalase enzyme with a novel affinity gel and investigation of its affinity against some pesticides

Catalase (E.C 1.11.1.1.6) is an antioxidant enzyme that breaks down hydrogen peroxide into water and oxygen. The enzyme plays an important role in oxidative stress by protecting cells against reactive oxygen species. The enzyme is found in virtually all aerobic organisms. Pesticides are chemicals used to increase agricultural productivity and control harmful organisms. These pesticides lead to water, soil and air pollution and affect living organisms. Therefore, some measures need to be taken to prevent the spread of pesticides into the environment and to control their effects. In this study, a new affinity gel (ω-aminohexyl agarose-3-amino-1,2,4-triazole-5-carboxylic acid) was synthesized to purify catalase enzyme from human erythrocytes. The parameters affecting the purification (pH, ionic strength, temperature) were optimized on the synthesized affinity gel. Catalase enzyme was purified 529.60-fold with 0.421% yield with the synthesized novel affinity gel. SDS-PAGE method was used to control enzyme purity and molecular mass. The molecular mass of the catalase enzyme was determined to be approximately 60 kDa. Km and Vmax kinetic constants of the enzyme were calculated for H2O2 and o-phenylenediamine substrate. In addition, the effects of fifteen different pesticides on catalase enzyme activity were investigated and Ki and inhibition types were determined.