Heat-induced aggregation of whey proteins: comparison of cheese WPC with acid WPC and relevance of mineral composition

Heat-induced aggregation of whey proteins in solutions made from two commercial whey protein concentrates (WPCs), one derived from mineral acid whey (acid WPC) and the other from cheese whey (cheese WPC), was studied using polyacrylamide gel electrophoresis (PAGE), size exclusion chromatography (SEC), and transmission electron microscopy (TEM). Heat treatment (75 degrees C) of acid WPC solutions (12.0%, w/w, pH 6.9) resulted in formation of relatively small "soluble" aggregates that were predominantly disulfide-linked. By contrast, heat treatment of the cheese WPC solutions (under the same conditions) caused formation of relatively large aggregates, containing high proportions of aggregates linked by noncovalent associations. The rate of aggregation of both beta-lactoglobulin and alpha-lactalbumin at 75 degrees C, measured as the loss of native proteins by PAGE, was higher in the cheese WPC solution than in the acid WPC solution. Cross dialysis of the two WPC solutions resulted in alteration of the mineral composition of each WPC solution and reversing their heat-induced aggregation behavior. The results demonstrated that the mineral composition is very important in controlling the aggregation behavior of WPC products.     

Pressure-induced unfolding and aggregation of the proteins in whey protein concentrate solutions

Whey protein concentrate solutions (12% w/v, pH 6.65 +/- 0.05) were pressure treated at 800 MPa for 20-120 min and then examined using size exclusion chromatography (SEC), small deformation rheology, transmission electron microscopy, and various types of one-dimensional (1D) and two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE). The pressure-treated samples showed a time-dependent loss of native whey proteins by SEC and 1D PAGE and a corresponding increase in non-native proteins and protein aggregates of different sizes. These aggregates altered the viscosity and opacity of the samples and were shown to be cross-linked by intermolecular disulfide bonds and by noncovalent interactions using 1D PAGE [alkaline (or native), sodium dodecyl sulfate (SDS), and SDS of reduced samples (SDS(R))] and 2D PAGE (native:SDS and SDS:SDS(R)). The sensitivity of the major whey proteins to pressure was in the order beta-lactoglobulin B (beta-LG B) > beta-LG A > bovine serum albumin (BSA) > alpha-lactalbumin (alpha-LA), and the large internal hydrophobic cavity of beta-LG may have been partially responsible for its sensitivity to high-pressure treatments. It seemed likely that, at 800 MPa, the formation of a beta-LG disulfide-bonded network preceded the formation of disulfide bonds between alpha-LA or BSA and beta-LG to form multiprotein aggregates, possibly because the disulfide bonds of alpha-LA and BSA are less exposed than those of beta-LG either during or after pressure treatment. It may be possible that intermolecular disulfide bond formation occurred at high pressure and that hydrophobic association became important after the high-pressure treatment.     

Effects of adding low levels of a disulfide reducing agent on the disulfide interactions of β-lactoglobulin and κ-casein in skim milk

Low concentrations of a disulfide reducing agent were added to unheated and heated (80 °C for 30 min) skim milk, with and without added whey protein. The reduction of the β-lactoglobulin and κ-casein disulfide bonds was monitored over time using electrophoresis. The distribution of the proteins between the colloidal and serum phases was also investigated. κ-Casein disulfide bonds were reduced in preference to those of β-lactoglobulin in both unheated and heated skim milk (with or without added whey protein). In addition, in heated skim milk, while the serum κ-casein was reduced more readily than the colloidal κ-casein, the distribution of κ-casein between the two phases was not affected.     

Formation of new protein structures in heated mixtures of BSA and alpha-lactalbumin

The heat-induced protein-protein interactions of alpha-lactalbumin (alpha-La) and bovine serum albumin (BSA), dispersed in a pH 6.8, 10% whey protein concentrates (WPC) permeate, were followed using alkaline and sodium dodecyl sulfate (SDS) 1D and 2D polyacrylamide gel electrophoresis (PAGE) and size-exclusion high-performance liquid chromatography (SE-HPLC). Heated (75 degrees C) 5% BSA solution contained large disulfide-bonded BSA aggregates, although some monomer BSA (SDS-monomeric BSA) could be dissociated from the aggregates by SDS. In contrast, similarly heated alpha-La solutions contained small quantities of several monomeric forms of alpha-La and dimeric alpha-La but no large aggregates. When 10% solutions of 1:1 (w/w) mixtures of alpha-La and BSA were heated, large disulfide-bonded aggregates and SDS-monomeric BSA and alpha-La were present. However, heated 2% mixtures contained more modified alpha-La monomers, alpha-La-dimers, and alpha-La-trimers, fewer large disulfide-bonded aggregates, and less SDS-monomeric alpha-La or BSA. These results suggest that BSA forms disulfide-bonded aggregates that contain available thiol groups that can catalyze the formation of differently structured alpha-La monomers, dimers, higher polymers, and adducts of alpha-La with BSA.     

Body mass index, physical activity and dietary behaviours among adolescents in the Kingdom of Tonga

OBJECTIVES: To compare the prevalence of overweight and obesity among adolescents using international and ethnic-specific cut-off points and to examine patterns of physical activity and dietary behaviours. DESIGN: Cross-sectional analytical study. SETTING: Schools on Tongatapu, Vava'u and Hapa'ai islands in the Kingdom of Tonga. SUBJECTS: A total of 443 school students aged 11-16 years underwent anthropometric measures of height and weight and provided self-reported measures of physical activity and dietary behaviours.Results Mean body mass index (BMI) was higher among girls than boys (23.7 kg m(-2) vs. 21.8 kg m(-2) and tended to increase with age. A total of 36.0% of boys and 53.8% of girls were overweight or obese using the international cut-off points, whereas 25.0% of boys and 37.6% of girls were classified in this way using Polynesian-specific cut-off points. Tinned mutton or beef was the food that most participants (56.9%) reported eating once or more per day. Over half of the young people did not eat taro, fruit or vegetables at least once per day. Regular physical activity outside of school hours was reported by 20.7% of respondents, and 58.2% watched 1 h or more of television per day. Physical activity participation was the only behaviour independently associated with a lower risk of overweight or obesity.Conclusions Using Polynesian-specific cut-off points for overweight and obesity the prevalence of these conditions was still among the highest found in adolescents. The prevalence of physical inactivity and poor dietary habits indicate that risk factors for chronic disease are well established during adolescence in Tonga.     

Behavior of protein in the presence of calcium during heating of whey protein concentrate solutions

The effect of added CaCl(2) on heat-induced changes in whey protein (WP) solutions prepared from whey protein isolate (WP1), acid whey protein concentrate (WP2), and cheese whey protein concentrate (WP3) was investigated. The loss of native-like, proteins, aggregation, and gel firmness of WP were maximum at certain levels of added CaCl(2). These levels were different for different WP products. The effect of added CaCl(2) on these changes appeared to be related to the initial calcium concentrations of these solutions. The higher the calcium content of the product, the less available sites for added CaCl(2) to bind. It was considered that addition of CaCl(2) changed the types of protein interactions that formed the protein aggregates during heating. Added calcium caused dramatic decreases in fracture stress of WP gels due to the formation of large protein aggregates.