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Thermodynamic characterization of calcium-milk protein interaction by isothermal titration calorimetry

Author:
Canabady-Rochelle, Latha-Selvi, Sanchez, Christian, Mellema, Michel, Banon, Sylvie
Source:
Dairy science & technology 2009 v.89 no.3-4 pp. 257-267
ISSN:
1958-5586
Subject:
calcium, calcium chloride, calorimetry, casein, cows, dairy protein, electrophoresis, electrostatic interactions, heat production, hydrophobicity, ions, micelles, milk, models, particle size, titration
Abstract:
In milk, casein micelles are the natural vectors of calcium (Ca) and constitute an ideal vehicle for the delivery of additional Ca, through Ca-protein interactions. Isothermal titration calorimetry (ITC) is a technique of choice in the study of biomolecular interactions and especially in mineral-protein interactions. Calcium-milk protein interactions were thermodynamically characterized by titration of cow’s milk protein with calcium chloride (CC) using ITC. Experiments similar to that of ITC were reproduced and analyzed with other techniques (granulometry and electrophoretic mobility measurement) to better understand the nature of interactions. Binding titration curves were fitted with the “one set of sites model” to determine the thermodynamic parameters (N, 6 mol Ca·mol⁻¹ of cow’s milk protein; K, 1070 mol·L⁻¹; ΔH, 1910 cal·mol⁻¹ of CC; and ΔS, cal·mol⁻¹·K⁻¹). Milk proteins bound up to 8 mg Ca·g⁻¹ of protein at saturation. The global thermodynamic signal obtained upon titration was endothermic. The electrophoretic mobility variations of casein micelles occurring upon CC titration were indicative of Ca-protein interactions of electrostatic nature. Hence, the usual exothermic signal involved in electrostatic interactions was completely hidden by the strong endothermic signal coming from the release of water molecules, either from the hydration shell of the Ca ions and/or dehydration of hydrophobic core of proteins. Particle size variations were indicative of casein micelle retraction upon CC titration.
Agid:
5483062