Covalent modification of glucose oxidase from Aspergillus niger by the palmitic acid ester of N-hydroxysuccinimide at a molar ratio ester:protein of 56:1 results in the formation of the enzyme derivative with 11 attached palmitic chains. Surface hydrophobicity measurements by a fluorescent probe, 8-anilino-1-naphthalenesulfonate, indicate a drastic increase in the hydrophobicity index of glucose oxidase after such a modification. The modified glucose oxidase displays a much higher adsorption affinity for hydrophilic (silica) as well as for hydrophobic (silica coated by phosphatidyl choline and cholesterol monolayers and polystyrene latex beads) surfaces, and forms more compact surface layers compared to the native glucose oxidase. Such a difference results from a spontaneous formation of micelle-like aggregates (clusters) of the hydrophobized enzyme molecules (average size 500 nm), which come into contact with a surface. A possible structure of the glucose oxidase surface layers and the nature of the forces determining the adsorption of the enzyme on various adsorbents are discussed.
- Glucose oxidase
- Lipid monolayer