Abstract
BackgroundActivity retention upon enzyme adsorption on inorganic nanostructures depends on different system parameters such as structure and composition of the support, composition of the medium as well as enzyme loading. Qualitative and quantitative characterization work, which aims at an elucidation of the microscopic details governing enzymatic activity, requires well-defined model systems.ResultsVapor phase-grown and thermally processed anatase TiO2 nanoparticle powders were transformed into aqueous particle dispersions and characterized by dynamic light scattering and laser Doppler electrophoresis. Addition of β-galactosidase (β-gal) to these dispersions leads to complete enzyme adsorption and the generation of β-gal/TiO2 heteroaggregates. For low enzyme loadings (~4% of the theoretical monolayer coverage) we observed a dramatic activity loss in enzymatic activity by a factor of 60–100 in comparison to that of the free enzyme in solution. Parallel ATR-IR-spectroscopic characterization of β-gal/TiO2 heteroaggregates reveals an adsorption-induced decrease of the β-sheet content and the formation of random structures leading to the deterioration of the active site.ConclusionsThe study underlines that robust qualitative and quantitative statements about enzyme adsorption and activity retention require the use of model systems such as anatase TiO2 nanoparticle agglomerates featuring well-defined structural and compositional properties.
Highlights
Activity retention upon enzyme adsorption on inorganic nanostructures depends on different system parameters such as structure and composition of the support, composition of the medium as well as enzyme loading
Since protein- and particle-related changes are strongly sensitive to a variety of chemical, physical and biological factors governing the interaction of the biomolecules with the inorganic material, enormous efforts have been devoted to the elucidation of related microscopic details [10,11,12,13,14,15]
Nanoparticle synthesis Anatase TiO2 nanocrystals were prepared by metal organic chemical vapor synthesis (MOCVS) based on the decomposition of titanium (IV) isopropoxide at T = 1073 K in a hot wall reactor system [28, 29]
Summary
Activity retention upon enzyme adsorption on inorganic nanostructures depends on different system parameters such as structure and composition of the support, composition of the medium as well as enzyme loading. Since protein- and particle-related changes are strongly sensitive to a variety of chemical, physical and biological factors governing the interaction of the biomolecules with the inorganic material, enormous efforts have been devoted to the elucidation of related microscopic details [10,11,12,13,14,15]. For this purpose more systematic and quantitative studies involving reference systems are needed [13]
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