A complex of ovotransferrin and lysozyme was directly isolated from egg white using an anti-transferrin antibody-immobilized membrane after antiserum proteins were separated by non-denaturing two-dimensional electrophoresis and transferred onto a membrane. The complex retained lysozyme activity that catalyzes the breakdown of peptidoglycans in the bacterial cell wall at the β1-4 bond between N-acetylmuramic acid and N-acetylglucosamine residues. The activity of the purified lysozyme was suppressed to 6.4% in the presence of 1 μmol Fe²⁺, whereas that of the mixture of the purified lysozyme and ovotransferrin was maintained at 58%. The activity of the purified lysozyme was suppressed to 35% in the presence of 10 nmol Fe³⁺, whereas that of the mixture of the purified lysozyme and ovotransferrin was maintained at 66%. Furthermore, the bacteriolytic activity against Bacillus subtilis of egg white with reduced glycoproteins such as ovotransferrin was assessed, and the bacteriolytic activity was found to be suppressed in the presence of Fe²⁺ and Fe³⁺. This suppression was ions, thereby alleviating the inhibition of lysozyme activity by iron ions. A complex of ovotransferrin and lysozyme is efficient because ovotransferrin effectively captures iron ions near lysozyme. Thus, protein complexes containing enzymes can be applied to control their activity.

Graphical Abstract