Abstract: The production of Fab’ fragment from specific egg yolk antibody against tyrosinase and its inhibitory activity on tyrosinase were investigated in this paper. The optimal conditions were obtained as follows: pH 4.0, ratio of egg yolk antibody to pepsin 1:150 (mg/U), and reaction time 8 h. Fab’ fragment purified by immunoaffinity column chromatography to a purity of over 95% exhibited good physicochemical stability. It was demonstrated as a non-competitive inhibitor with an IC50 value of 11.16 μmol/L and a binding constant of 5.79 × 105, whose inhibitory effect was much higher than that of IgY. The results of this study can provide a theoretical basis for the application of egg yolk antibody Fab’ fragments in food, cosmetics and pharmaceutical industries as a potential enzyme inhibitor.

Key words: tyrosinase, egg yolk antibody, enzymatic proteolysis, fragment antigen binding (Fab’ fragment), enzyme activity inhibition