Abstract

Renaturation of horseradish peroxidase from guainidine hydrochloride has been studied. Although refolding of the secondary structure was complete, only partial heme incorporation and recovery of enzymatic activity were observed. Heme capturing became less efficient at lower peroxidase concentrations: the refolding yield decreased from 60% at 1 μM to 10% at 0.1 μM concentration of the protein. Probing with conformation-sensitive antibodies indicated structural differences between peroxidase refolded at low concentration and the holo-enzyme.

Keywords: horseradish peroxidase, apo-enzyme, Refolding, heme, guanidine hydrochloride