An antioxidant material composed of halloysite nanotubes
(HNTs), protamine sulfate polyelectrolyte (PSP), and superoxide dismutase
(SOD) enzyme was prepared by self-assembly of the PSP and SOD biomacromolecules
on the nanoparticulate support. The structural, colloidal and biocatalytic
features were assessed. Adsorption of PSP on the oppositely charged
HNT surface at appropriate loadings gave rise to charge neutralization
and overcharging, which resulted in unstable and stable dispersions,
respectively. The formation of a saturated PSP layer on the HNT led
to the development of positive surface charge and to remarkable resistance
against salt-induced aggregation making the obtained HNT-PSP hybrid
suitable for immobilization of negatively charged SOD. No enzyme leakage
was observed from the HNT-PSP-SOD composite indicating sufficient
structural stability of this material due to electrostatic, hydrophobic,
and hydrogen bonding interactions taking place between the particles
and the biomacromolecules. Enzymatic assays revealed that SOD kept
its functional integrity upon immobilization and showed high activity
in superoxide radical dismutation. In this way, stable antioxidant
bionanocomposite dispersions were obtained, which can be used as antioxidants
in heterogeneous samples.