Purification and characterization of peroxidase enzyme from Sevketibostan (cnicus benedictus) plant

In this study, Peroxidase (POD) enzyme was purified from Şevketibostan (Cnicus benedictus) and its kinetic properties were investigated. First, POD enzyme was purified by the triple-phase separation technique (TPP) and its purity was checked by SDS-PAGE method. The molecular weight of the POD enzyme was found to be 63 kDa. In the kinetic characterization studies of the enzyme, 4-methyl catechol, H2O2, o-phenylene diamine, gallic acid, caffeic acid and pyrogallol substrates were used to determine the substrate specificity. Michaelis-Menten (Km) constant and maximum reaction rate (Vmax) values were calculated for all substrates used. In the characterization studies of the enzyme, optimum pH, optimum temperature, optimum ionic strength, salt tolerance and storage stability of the enzyme were investigated. POD enzyme from Sevketibostan (Cnicus benedictus) was found to have an optimum pH of 7.0 and an optimum temperature of 30 oC. Sevketibostan POD enzyme ascorbic acid, sodium azide (NaN3), Fe+3, Fe+2, Hg+2, Sn+2, Pb+2, Cd+2, Cu+2, K+, Na+, Ca+ metals, DMSO, DMF inhibits ethanol, acetone and t-butanol organic solvents.