Inactivation of urate oxidase by a system composed of lactoperoxidase, hydrogen peroxide and bromide

Urate oxidase from Candida utilis, an enzyme containing an essential thiol, was examined for its sensitivity to lactoperoxidase, an oxidant present in breast milk. Upon exposure to a system composed of lactoperoxidase, hydrogen peroxide and bromide at moderately alkaline pH, the urate oxidase exhibited comparable activity to the untreated enzyme; but upon exposure at moderately acidic pH, it lost its activity completely. Thus the lactoperoxidase-H2O2-bromide system significantly inactivated urate oxidase only at moderately acidic pH. This inactivation was prevented by the presence of N-acetylmethionine, a methionine analogue, or glutathione, which is a thiol compound analogous to an amino acid, indicating that it was probably due to the oxidation and damage of the methionine residue and/or the thiol group in the urate oxidase by the lactoperoxidase system, that loss of catalytic activity of the urate oxidase occurred.