Brewster angle microscopy of adsorbed protein films at air–water and oil–water interfaces after compression, expansion and heat processing

Recent Brewster angle microscopy (BAM) observations of adsorbed films of proteins at the air-water (A–W) and oil–water (O–W) interfaces are reviewed and compared. At the A–W interface β-lactoglobulin (β-L) and ovalbumin (OA) were studied at pH 7 and 5. At the O–W interface β-L, αs1-, β- and κ-caseins were studied at pH 7. The adsorbed films were periodically subjected to compression and expansion cycles such that the film area was typically varied between 125% and 50% of the original film area. At the A–W interface, little structure was observable on compression or expansion or aging, especially at pH 7. For ovalbumin at pH 5, some cracks and ridges in the films appeared. But, for both β-L and OA, such features became much more obvious on addition to the interface of a low area fraction (<0.01%) of 20 μm polystyrene latex (PS) particles. With particles present the structuring was also more obvious at pH 5 (closer to the protein isoelectric point) than at pH 7, and for greater adsorption times and/or higher bulk protein concentration (Cb). Particle addition was not necessary to highlight folding and ridges that occurred at the O–W interface for β-L. After heating to 80 and 90 °C, β-L films adsorbed from low Cb (0.005 wt%) even greater film structuring was evident. However, β-L films adsorbed from higher Cb (≥0.05 wt%) showed fewer, but more pronounced ridges and cracks. The caseins at the O–W interface showed comparatively little evidence of structuring, either before or after heating. A measure of the dilatational elastic modulus of the films correlated with the observed variations in the structural integrity of the films. Clearly, protein films subjected to these types of thermal and mechanical perturbations can become highly inhomogeneous, depending on the type of protein and the bulk solution conditions, with implications for the stability of the corresponding foams and emulsions. This is in agreement with earlier computer simulations. Protein films at the O–W interface (particularly after heating) appear to be more resilient and less aggregated than films at the A–W interface.