Stabilizing and Suicide-Peroxide Protecting Effect of Ni2+ on Horseradish Peroxidase

The present research discusses the structure stabilizing and protecting effects of Ni2+ against suicide-peroxide inactivation of horseradish peroxidase (HRP). Suicide inactivation of HRP by hydrogen peroxide (3 mM) was monitored by measuring change in the absorbance of the colored product (tetraguaiacol) of the catalytic reaction cycle at 470 nm. Progress curves of the catalytic reaction cycle were obtained at 27 oC, phosphate buffer (5 mM), pH 7.0. The corresponding kinetic parameters (e.g., initial enzyme activity (ao) and the apparent rate constant (ki) of suicide inactivation of HRP by peroxide) were evaluated using a kinetic equation derived in this study. Comparative activatory and inhibitory effects of Ni2+ on the kinetics of suicide-peroxide inactivation of HRP are discussed.