Kinetics of formation and physicochemical characterization of thermally-induced beta-lactoglobulin aggregates.
The kinetics of heat denaturation and aggregation for beta-lactoglobulin dispersions (5% w/v) were studied at 3 pHs (6, 6.4, and 6.8) and at a heating temperature of 80 degrees C. Protein aggregates were characterized for hydrodynamic diameter, microstructure, and molecular weight by means of dynamic light scattering, transmission electron microscopy, and polyacrylamide gel electrophoresis, respectively. Concentration of native beta-lactoglobulin decreased with holding time and with a decrease in the pH. Apparent rate constants were calculated for beta-lactoglobulin denaturation applying the general kinetic equation solved for a reaction order of 1.5. Values of the apparent reaction rate constant k = 7.5, 6.3 and 5.6 x 10(-3) s(-1) were found for pH 6, 6.4, and 6.8, respectively. Decreasing the pH of the dispersions produced higher aggregate sizes. After a holding time of 900 s, average hydrodynamic diameters for beta-lactoglobulin aggregates at pH 6, 6.4, and 6.8 were 96, 49, and 42 nm, respectively. These results were confirmed by transmission electron microscopy images, where a shift in the size and morphology of aggregates was found, from large and spherical at pH 6 to smaller and linear aggregates at pH 6.8. beta-Lactoglobulin formed disulfide-linked intermediates (dimers, trimers, tetramers) and so on) which then formed high molecular weight aggregates. From the results obtained by DLS, TEM, and SDS-PAGE a mechanism for beta-lactoglobulin aggregation was proposed. This study shows that heat treatment can be used to produce protein aggregates with different sizes and morphologies to be utilized as ingredients in foods.
Zúñiga
R N
RN
Dept. of Chemical & Bioprocess Engineering, Pontificia Univ. Católica de Chile, Avenida Vicuña Mackenna 4860, Macul, Santiago, Chile. rnzuniga@uc.cl
Tolkach
A
A
Kulozik
U
U
Aguilera
J M
JM
D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000032:analysis / Q000737:chemistry*; D046529:Microscopy, Electron, Transmission / Q000379:methods; D010316:Particle Size; D011489:Protein Denaturation; D013499:Surface Properties; D013816:Thermodynamics
D007782:Lactoglobulins