1- hidroksipireno peroksidazinės oksidacijos tyrimas

Abstract

The oxidation of 1-hydroxypyrene (1-HP) catalyzed with re combinant Coprinus cinereus (rCiP) peroxidase was investigated. The high fluorescence quantum yield (Of = 0.66) for 1-HP and the high bimolecular constant of 1-HP oxidation (kox = (1.1 ± 0,4) • 108 M 's ') allowed using this reaction for determination of submicro- molar hydrogen peroxide concentration. The hydrogen peroxide de- termination analysis was performed at pH 5.0-8.0. The established hydrogen peroxide detection limit is equal to 0.6 nM with pH 8.0. For comparison the same experiments as with 1-HP were carried out with Amplex Red (AR) compound which is a patent reagent frequently used for low hydrogen peroxide concentration detection. In order to explain the achieved results the mathematical modeling and optimization of 1-HP and AR initial oxidation rate dependence on hydrogen peroxide concentration and pH in stationary conditions was performed. The resembling results have proved the superiority of 1-HP peroxidase oxidation for determination of submicromolar hydrogen peroxide concentration.