Covalent immobilization of lysozyme on stainless steel. Interface spectroscopic characterization and measurement of enzymatic activity

A new strategy aiming at the protection of metallic surfaces against the growth of biofilms is presented here. This work reports the grafting of primary amines by aminosilanization of oxidized stainless steel followed by chemical coupling of the glycosidase lysozyme from hen egg white using glutaraldehyde as homobifunctional cross-linking agent. Controlled characterization of a stainless steel surface by X-ray photoelectron spectroscopy and Fourier transform infrared reflection-absorption spectroscopy at each step enabled the mode of binding, coverage, and orientation of the grafted molecules to be addressed. As a result, the stainless steel samples covered with a covalently immobilized layer of lysozyme showed some lytic activity on a suspension of bacteria Micrococcus lysodeikticus.