Purification, characterization, and inhibition sensitivity of peroxidase from wheat (Triticum aestivum ssp vulgare)

The purification and characterization of peroxidase is currently growing interests since peroxidases have implications in various industrial and biochemical applications. In this study, wheat peroxidase was purified using (NH4)(2)SO4 precipitation, CM-Sephadex cation exchange, and gel filtration chromatographies. Enzyme purity and molecular mass were checked by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Enzyme kinetics was studied using two substrates: guaiacol and hydrogen peroxide (H2O2). K-m and V-max values were calculated from Lineweaver-Burk graph for each substrate. Wheat peroxidase activity has been enhanced 284-fold. Wheat peroxidase had molecular mass of 38.8 kDa. K-m values for guaiacol and H2O2 were found as 2.467 mM and 7.307 mM, respectively. The pH and temperature optima were 5.5 and 40.0 degrees C, respectively. Also, the enzyme was highly inhibited by citric acid and Cetyltrimethylammonium bromide.