pH-Dependent behaviour of soluble protein aggregates formed during heat-treatment of milk at pH 6.5 or 7.2.
The pH-dependent behaviour of soluble protein aggregates produced by the pre-heating of reconstituted skim milk at 90 degrees 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 degrees C or GDL at 20 and 38 degrees 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 degrees 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 kappa-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.
Renan
Marie
M
UMR 1253 Science et Technologie du Lait et de l'Oeuf, Inra-Agrocampus Rennes, 65 rue de St-Brieuc, 35042 Rennes Cedex, France.
Mekmene
Omar
O
Famelart
Marie-Hélène
MH
Guyomarc'h
Fanny
F
Arnoult-Delest
Véronique
V
Pâquet
Denis
D
Brulé
Gérard
G
D000818:Animals; D011232:Chemical Precipitation; D002845:Chromatography / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005782:Gels / Q000737:chemistry; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008823:Micelles; D008892:Milk / Q000737:chemistry*; D008894:Milk Proteins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D010316:Particle Size; D012995:Solubility
D005782:Gels; D008823:Micelles; D008894:Milk Proteins