Particle size might affect the inhibition behaviors of
gold nanoparticles
(AuNPs) on enzyme activity by influencing the density of binding sites
(ρ), the association constant (K
a), the steric hindrance of enzymes by AuNPs, the binding
orientations of the enzyme on AuNPs, as well as the structural changes
of enzymes. In previous studies, the effects of the above-mentioned
factors, which could not be ignored in the applications of enzymatic
electrochemistry, were often overshadowed by the effects of surface
area. In order to study the size effect on the inhibition types and
inhibitory ability of enzymes by AuNPs, we investigated the inhibition
behaviors of chymotrypsin (ChT) by AuNPs with three different sizes
(D1-AuNCs, D3-AuNPs, and D6-AuNPs) under the same surface area concentration.
The results showed that both of the inhibition types and the inhibition
ability varied with the particle size of AuNPs. D1-AuNCs inhibited
ChT noncompetitively, while D3/D6-AuNPs inhibited ChT competitively.
Contrary to the common sense, D6-AuNPs showed a weaker inhibitory
ability than D3-AuNPs. By means of zeta potential, agarose gel electrophoresis,
isothermal titration calorimetry, synchronous fluorescence spectroscopy,
and circular dichroism, the mechanism of the weak inhibitory ability
of D6-AuNPs was found to be the standing binding orientation caused
by the small curvature. This work had certain guiding significance
for the biosafety of AuNPs, the development of nanoinhibitors, as
well as the applications of AuNPs in enzymatic electrochemistry.