Effects of Aqueous Isotopic Substitution on the Adsorption Dynamics and Dilational Rheology of β-Lactoglobulin Layers at the Water/Air Interface

Abstract

The effect of the degree of isotopic substitution of the aqueous medium on the adsorption kinetics and the surface dilational rheological behavior at the water/air interface of the globular protein β-lactoglobulin was investigated. Aqueous solutions with fixed concentrations of 1 μM protein and 10 mM hydrogenous buffer with controlled pH 7 were prepared in H 2 O, D 2 O, and an isotopic mixture of 8.1% v/v D 2 O in H 2 O (called air contrast matched water, ACMW). Using a bubble shape analysis tensiometer, we obtained various experimental dependencies of the dilational viscoelasticity modulus E as a function of the dynamic surface pressure and of the frequency and amplitude of bubble surface area oscillations, either in the course of adsorption or after having reached a steady state. In general, the results revealed virtually no effect from substituting H 2 O by ACMW but distinct albeit relatively weak effects for intermediate adsorption times for D 2 O as the aqueous phase. In the final stage of adsorption, established after around 10 h, the equilibrium adsorption and the dilational rheological behavior of all protein layers under investigation are only very weakly affected by the presence of D 2 O. The obtained results help to design experimental protocols for protein adsorption studies, for example, by neutron reflectivity.