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Kinetic study of peroxidase-catalyzed oxidation of 1-hydroxypyrene. Development of a nanomolar hydrogen peroxide detection system

Kinetics of 1-hydroxypyrene (1-HP) oxidation catalyzed with recombinant Coprinus cinereus (rCiP) and horseradish (HRP) peroxidases was investigated with a special emphasis for developing a nanomolar hydrogen peroxide (H2O2) detection system. At pH 8.0 the bimolecular constants of 1-HP oxidation with the ferryl compounds of rCiP and HRP were equal to (1.0 ± 0.3) × 108 M−1 s−1 and (0.6 ± 0.2) × 108 M−1 s−1, respectively. High bimolecular constants and fluorescence quantum yield of 1-HP (0.66) permitted detection as low as 21 nM of H2O2. To optimize the detection system 1-HP oxidation was modeled at steady-state conditions in the range pH 5.0 to pH 8.0. The 1-HP based detection system was compared with the Amplex Red system. The peroxidase-catalyzed 1-HP oxidation system was used for determination of ozone in the air.