Effect of antioxidants (digoxin, quercetin, and ascorbic acid) on catalytic properties of horseradish peroxidase.

Antioxidants (digoxin, quercetin, and ascorbic acid) inhibited the oxidation of o-dianisidine catalyzed by horseradish peroxidase. Digoxin bound with the enzyme-substrate complex which included a stable semioxidized product of o-dianisidine and inhibited horseradish peroxidase by the anticompetitive pattern, while the enzyme inhibition by quercetin followed the mixed pattern. The oxidation of ascorbic acid and o-dianisidine with their combined presence in the reaction mixture was differentiated. o-Dianisidine was oxidized after the oxidation of 90-100% of ascorbic acid was completed. The rate of peroxidase oxidation of ascorbic acid in the presence of o-dianisidine was more than two orders higher than the rate of its individual oxidation and 1.5-2.0 times higher than the rate of o-dianisidine oxidation. Feasible inhibition mechanisms of peroxidase oxidation of o-dianisidine are discussed.