pH-Dependent behaviour of soluble protein aggregates formed during heat-treatment of milk at pH 6·5 or 7·2

Abstract

The pH-dependent behaviour of soluble protein aggregates produced by the pre-heating of reconstituted skim milk at 90 °C for 10 min was studied, in order to understand the role of these aggregates in acid gelation of heated milk. The following milk samples were prepared: (1) control (unheated reconstituted milk, pH 6·5); (2) milk heat-treated at pH 6·5 (mHtd6·5) and (3) milk heat-treated at pH 7·2 (mHtd7·2). They were centrifuged and the supernatants (SPNT 1) pH-adjusted to yield a series of pH values ranging from 6·5 or 7·2 to 4·6 using HCl at 20 °C or GDL at 20 and 38 °C. pH-Adjusted SPNTs 1 were re-centrifuged. The resulting supernatants (SPNTs 2) were analysed by OD (at 600 and 280 nm) and SDS-PAGE in order to characterise proteins still soluble as a function of pH. Particle size in SPNTs 1 was analysed by Steric Exclusion Chromatography. The OD600 nm revealed that during acidification soluble casein in both control and heat-treated samples exhibits variations in its optical properties or size as previously shown with micellar casein. In heat-treated samples, soluble casein and heat-induced covalent soluble aggregates precipitate at the same pH value. A progressive acidification of the soluble phase did not separate them. Increasing the temperature of acidification from 20 to 38 °C resulted in an increase in the precipitation pH of the proteins. However choice of acidifier did not have a significant effect on OD profiles. The soluble covalent aggregates from mHtd7·2 were smaller, more numerous, and had a higher content of κ-casein than mHtd6·5. Both types of aggregates began to precipitate at the same pH value but precipitation occurred over a narrower pH-range for soluble aggregates prepared from mHtd7·2. This may explain the higher gelation pH of mHtd7·2 compared with mHtd6·5.