Interactions between whey proteins and salivary proteins as related to astringency of whey protein beverages at low pH

Whey protein beverages have been shown to be astringent at low pH. In the present study, the interactions between model whey proteins (β-lactoglobulin and lactoferrin) and human saliva in the pH range from 7 to 2 were investigated using particle size, turbidity, and ζ-potential measurements and sodium dodecyl sulfate-PAGE. The correlation between the sensory results of astringency and the physicochemical data was discussed. Strong interactions between β-lactoglobulin and salivary proteins led to an increase in the particle size and turbidity of mixtures of both unheated and heated β-lactoglobulin and human saliva at pH ∼3.4. However, the large particle size and high turbidity that occurred at pH 2.0 were the result of aggregation of human salivary proteins. The intense astringency in whey protein beverages may result from these increases in particle size and turbidity at these pH values and from the aggregation and precipitation of human salivary proteins alone at pH <3.0. The involvement of salivary proteins in the interaction is a key factor in the perception of astringency in whey protein beverages. At any pH, the increases in particle size and turbidity were much smaller in mixtures of lactoferrin and saliva, which suggests that aggregation and precipitation may not be the only mechanism linked to the perception of astringency in whey protein.     

Roles of charge interactions on astringency of whey proteins at low pH

Whey proteins are a major ingredient in sports drink and functional beverages. At low pH, whey proteins are astringent, which may be undesirable in some applications. Understanding the astringency mechanism of whey proteins at low pH could lead to developing ways to minimize the astringency. This study compared the astringency of β-lactoglobulin (β-LG) at low pH with phosphate buffer controls having the same amount of phosphate and at similar pH. Results showed that β-LG samples were more astringent than phosphate buffers, indicating that astringency was not caused by acid alone and that proteins contribute to astringency. When comparing among various whey protein isolates (WPI) and lactoferrin at pH 3.5, 4.5, and 7.0, lactoferrin was astringent at pH 7.0 where no acid was added. In contrast, astringency of all WPI decreased at pH 7.0. This can be explained by lactoferrin remaining positively charged at pH 7.0 and able to interact with negatively charged saliva proteins, whereas the negatively charged WPI would not interact. Charge interactions were further supported by β-LG or lactoferrin and salivary proteins precipitating when mixed at conditions where β-LG, lactoferrin, or saliva themselves did not precipitate. It can be concluded that interactions between positively charged whey proteins and salivary proteins play a role in astringency of proteins at low pH.     

Consumer perception of astringency in clear acidic whey protein beverages

Acidic whey protein beverages are a growing component of the functional food and beverage market. These beverages are also astringent, but astringency is an expected and desirable attribute of many beverages (red wine, tea, coffee) and may not necessarily be a negative attribute of acidic whey protein beverages. The goal of this study was to define the consumer perception of astringency in clear acidic whey protein beverages. Six focus groups (n=49) were held to gain understanding of consumer knowledge of astringency. Consumers were presented with beverages and asked to map them based on astringent mouthfeel and liking. Orthonasal thresholds for whey protein isolate (WPI) in water and flavored model beverages were determined using a 7-series ascending forced choice method. Mouthfeel/basic taste thresholds were determined for WPI in water. Acceptance tests on model beverages were conducted using consumers (n=120) with and without wearing nose clips. Consumers in focus groups were able to identify astringency in beverages. Astringency intensity was not directly related to dislike. The orthonasal threshold for WPI in water was lower (P < 0.05) than the mouthfeel/basic taste threshold of WPI in water. Consumer acceptance of beverages containing WPI was lower (P < 0.05) when consumers were not wearing nose clips compared to acceptance scores of beverages when consumers were wearing nose clips. These results suggest that flavors contributed by WPI in acidic beverages are more objectionable than the astringent mouthfeel and that both flavor and astringency should be the focus of ongoing studies to improve the palatability of these products.     

酒度、总酸、pH值以及饮用温度对干红葡萄酒涩味的影响

利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulphate polyacrylamide gel electrophoresis,SDS-PAGE）分析唾液蛋白与模拟酒反应后蛋白减少比例,并将其表示为涩味强度;同时,以酒度、总酸、pH值以及饮用温度为考察因素,利用二次正交旋转组合设计分析各因子对涩味强度的影响。结果表明：pH值对涩度影响最大,其次是酸度和温度,酒度影响最小;其中pH值和酸度互作效应对涩味的影响显著。     

Ready-to-drink protein beverages: Effects of milk protein concentration and type on flavor

This study evaluated the role of protein concentration and milk protein ingredient [serum protein isolate (SPI), micellar casein concentrate (MCC), or milk protein concentrate (MPC)] on sensory properties of vanilla ready-to-drink (RTD) protein beverages. The RTD beverages were manufactured from 5 different liquid milk protein blends: 100% MCC, 100% MPC, 18:82 SPI:MCC, 50:50 SPI:MCC, and 50:50 SPI:MPC, at 2 different protein concentrations: 6.3% and 10.5% (wt/wt) protein (15 or 25 g of protein per 237 mL) with 0.5% (wt/wt) fat and 0.7% (wt/wt) lactose. Dipotassium phosphate, carrageenan, cellulose gum, sucralose, and vanilla flavor were included. Blended beverages were preheated to 60°C, homogenized (20.7 MPa), and cooled to 8°C. The beverages were then preheated to 90°C and ultrapasteurized (141°C, 3 s) by direct steam injection followed by vacuum cooling to 86°C and homogenized again (17.2 MPa first stage, 3.5 MPa second stage). Beverages were cooled to 8°C, filled into sanitized bottles, and stored at 4°C. Initial testing of RTD beverages included proximate analyses and aerobic plate count and coliform count. Volatile sulfur compounds and sensory properties were evaluated through 8-wk storage at 4°C. Astringency and sensory viscosity were higher and vanillin flavor was lower in beverages containing 10.5% protein compared with 6.3% protein, and sulfur/eggy flavor, astringency, and viscosity were higher, and sweet aromatic/vanillin flavor was lower in beverages with higher serum protein as a percentage of true protein within each protein content. Volatile compound analysis of headspace vanillin and sulfur compounds was consistent with sensory results: beverages with 50% serum protein as a percentage of true protein and 10.5% protein had the highest concentrations of sulfur volatiles and lower vanillin compared with other beverages. Sulfur volatiles and vanillin, as well as sulfur/eggy and sweet aromatic/vanillin flavors, decreased in all beverages with storage time. These results will enable manufacturers to select or optimize protein blends to better formulate RTD beverages to provide consumers with a protein beverage with high protein content and desired flavor and functional properties.     

美国乳清蛋白在即饮饮料中的应用

乳清在乳制品加工中的应用前景非常广阔,本刊特邀美国乳品出口协会合作开辟专栏,就乳清在食品中应用的最新资讯进行专题系列报道。     

Ability of whey protein isolate and/or fish gelatin to inhibit physical separation and lipid oxidation in fish oil-in-water beverage emulsion

The effect of pH on the capability of whey protein isolate (WPI) and fish gelatin (FG), alone and in conjugation, to form and stabilize fish oil-in-water emulsions was examined. Using layer-by-layer interfacial deposition technique for WPI–FG conjugate, a total of 1% protein was used to prepare 10% fish oil emulsions. The droplets size distributions and electrical charge, surface protein concentration, flow and dynamic rheological properties and physiochemical stability of emulsions were characterize at two different pH of 3.4 and 6.8 which were selected based on the ranges of citrus and milk beverages pHs, respectively. Emulsions prepared with WPI–FG conjugate had superior physiochemical stability compare to the emulsions prepared with individual proteins. Higher rate of coalescence was associated with reduction in net charge and consequent decrease of the repulsion between coated oil droplets due to the proximity of pH to the isoelectric point of proteins. The noteworthy shear thinning viscosity, as an indication of flocculation onset, was associated with whey protein stabilized fish oil emulsion prepared at pH of 3.4 and gelatin stabilized fish oil emulsion made at pH of 6.8. At pH 3.4, it appeared that lower surface charge and higher surface area of WPI stabilized emulsions promoted lipid oxidation and production of hexanal.     

Astringency of bovine milk whey protein

Whey protein solutions at pH 3.5 elicited an astringent taste sensation. The astringency of whey protein isolate (WPI), the process whey protein (PWP) that was prepared by heating WPI at pH 7.0, and the process whey protein prepared at pH 3.5 (aPWP) were adjusted to pH 3.5 and evaluated by 2 sensory analyses (the threshold method and the scalar scoring method) and an instrumental analysis (taste sensor method). The taste-stimulating effects of bovine and porcine gelatin were also evaluated. The threshold value of astringency of WPI, PWP, and aPWP was 1.5, 1.0, and 0.7 mg/mL, respectively, whereas the gelatins did not give definite astringency. It was confirmed by the scalar scoring method that the astringency of these proteins increased with the increase in protein concentration, and these proteins elicited strong astringency at 10 mg/mL under acidic conditions. On the other hand, the astringency was not elicited at pH 3.5 by 2 types of gelatin. A taste sensor gave specific values for whey proteins at pH 3.5, which corresponded well to those obtained by the sensory analysis. Elicitation of astringency induced by whey protein under acidic conditions would be caused by aggregation and precipitation of protein molecules in the mouth.     

Correlations between saliva protein composition and some T–I parameters of astringency

Saliva samples were collected from 12 panellists immediately before and after the sensory assessment of two red wine samples and the salivary proteins in all samples were analysed by high performance liquid chromatography (HPLC). Three peaks appeared in the majority of the chromatograms and the areas of these peaks were individually correlated for each assessor against four astringency time–intensity (T–I) parameters (time to maximum intensity, total duration, maximum intensity and area under the T–I curve). Astringency was not correlated with the total area in the saliva chromatograms. However, statistically significant correlations were obtained between the area of particular proteins and the sensory data indicating that the concentration of individual proteins in saliva might be more important for astringency than the total protein content. Significant correlations were also obtained between the relative concentration of individual proteins and the T–I data.     

The effect of protein profile and preheating on denaturation of whey proteins and development of viscosity in milk protein beverages during heat treatment

The effect of preheat temperature (63 or 77 °C for 30 s; final heat 120 °C for 30 s) and casein to whey protein ratio on the physical characteristics of 3.3%, w/w, dairy protein beverages was investigated. Dispersions preheated at 77 °C had lower viscosity than dispersions preheated at 63 °C. Casein‐containing dispersions had significantly lower levels of α‐lactalbumin denaturation than whey protein‐only dispersions. A higher proportion of casein improved the thermal stability of protein dispersions. Overall, alteration of preheat temperature and casein to whey protein ratio can influence dairy beverage quality, with increasing levels of casein reducing physical changes due to heat treatment.     

Astringency sub-qualities of red wines and the influence of wine–saliva aggregates

Astringency is a sensory attribute, related to the quality and mouthfeel of red wines. However, the origin of astringency sub-qualities, such as the typical drying astringency found in immature grapes, is still unknown. Astringency of red wines with similar tannin content but different astringency sub-qualities, from different harvest dates, is studied. Astringency was characterised in terms of friction coefficient, polyphenol content, sensory analysis and tannin/salivary–proteins aggregates characterisation. A different evolution during ripening was found for both Cabernet Sauvignon and Carménère, and tannin–protein aggregates showed differences in size, shape and surface. The velvety sub-quality appears to be related to aggregates with low precipitation, and with specific surface characteristics as roundness and Feret diameter. Results from this work propose an effect of aggregates on sensory perception and opens the possibility to explore their effect on oral lubrication.     

Formulation of apple juice beverages containing whey protein isolate or whey protein hydrolysate based on sensory and physicochemical analysis

Whey protein isolate (WPI) or its bioactive hydrolysate (WPH) was mixed with apple juice along with sweetener, obtaining a series of beverages with various pH values. Sedimentation of WPI‐apple juice and WPH‐apple juice beverages was inhibited at pH values of 3.15 and 3.47, respectively. The higher the whey protein content, the more undesirable was the taste of samples. A clearer appearance with smaller particle size was obtained with WPH‐apple juice formulations compared to WPI‐apple juice formulations at pH values closer to the pI of the whey proteins. Intrinsic viscosity measurements revealed the weaker associations of peptides compared with protein molecules.     

Turbidity and Protein Aggregation in Whey Protein Beverages

ABSTRACT:  During storage of heat-treated acidic (pH ≤ 4.6) whey protein beverages, formation of protein aggregates can create undesirable turbidity and sedimentation. In this study, we found that a slow protein aggregation process controlled the rate of formation of sediment and turbidity. A heat-treated model beverage containing 12.5 g/L whey protein at a pH of 4 was stored for 6 wk at 3 different storage temperatures and analyzed for turbidity, soluble protein, and protein aggregates. One sample was stored without further treatment and the other sample was filtered to remove protein aggregates formed during heating. This was done to test the hypothesis that aggregates formed during heat treatment served as nuclei for deposition of soluble protein. Turbidity increased and soluble protein decreased as protein aggregates formed during storage. Increasing the storage temperature accelerated this process. The loss of soluble protein during storage was fit to first- and second-order kinetic equations, allowing the prediction of the effect of protein concentration, storage time, and storage temperature on the formation of protein aggregates.     

Rheology of whey protein isolate-xanthan mixed solutions and gels. Effect of pH and xanthan concentration

Flow properties at pH 5.5-7.5 of whey protein isolate (WPI)-xanthan solutions containing 0-0.5 w/w% xanthan were studied by viscosimetry, although rigidity and fracture properties of the corresponding heat-set gels (90°C, 30 min) were determined by uniaxial compression. All the studied solutions displayed generalized shearthinning flow behaviour. Synergistic WPI-xanthan interactions has been revealed by observing that rheological parameters [σ_msf, K, n, η (γ)] characterizing blends were larger than those calculated from the two separated solutions. Such a behaviour was attributed to segregative phase separation of whey proteins and xanthan. Effects of xanthan on WPI-xanthan gel properties both depended on pH and xanthan concentration. Simultaneous increased xanthan concentration and decreased pH inhibited gelation of WPI-xanthan blends. Regarding gel strength, synergistic WPI-xanthan interactions were observed at pH >7.0 and low xanthan concentration (0.05 or 0.1 w/w%). Antagonism between the two macromolecules occurred at low xanthan concentration and pH ≤6.5, and high xanthan concentration (0.2 or 0.5 w/w%) at all pH tested. Low xanthan concentration rendered mixed gels more brittle than protein gels, and high xanthan concentration decreased pH effects on gel stress-strain relationships. The balance between strong thermal aggregation of concentrated whey proteins - in presence of incompatible xanthan -, high viscosity of blends and repulsive surface forces of protein molecules was thought to be at the origin of WPI-xanthan gel mechanical properties.     

Whey beverages decrease blood pressure in prehypertensive and hypertensive young men and women

Whey protein beverages reduced blood pressure in young men and women in a six week controlled intervention. There were no differences in systolic blood pressure (SBP), diastolic blood pressure (DBP), or mean arterial pressure (MAP) observed between groups consuming 28 g per day of either hydrolyzed or non-hydrolyzed whey protein in a beverage. However, in young adults with elevated DBP and SBP, whey beverage consumption significantly decreased SBP, DBP, and MAP by 8.0, 8.6, and 6.4 mm Hg, respectively (P ≤ 0.001 for all comparisons). In subjects with elevated SBP only, SBP significantly decreased by 3.8 mm Hg (P ≤ 0.04) after the whey beverage intervention. Subjects with normal blood pressure had no change in SBP, DBP, and MAP. Whey beverages also significantly decreased total and low-density lipoprotein cholesterol concentrations (P ≤ 0.001 and 0.05, respectively). Whey protein beverages may be useful for the dietary treatment of prehypertension and/or stage 1 hypertension.     

Effects of Calcium Addition on Stability and Sensory Properties of Soy Beverage

Pasteurized or thermally processed soy beverages (6% soy solids) were fortified to a comparable level of cow's milk with 25 mM (or 30 mM) calcium using mixtures of calcium citrate and tricalcium phosphate. These fortified pasteurized products had acceptable sensory properties. Addition of these calcium salts did not adversely affect protein stability of the beverage. Calcium citrate addition caused a decrease in beverage pH and viscosity. Thermally processed (still retort and agitort) canned beverages containing calcium salts were stable for 6 months when stored at 1°C or at room temperature.     

Interactions of enological tannins with the protein fraction of saliva and astringency perception are affected by pH

The effects of pH on both tannin-induced astringency and tannin–salivary protein interactions were investigated. A trained sensory panel evaluated astringency perception. Tannin–salivary protein interactions were assessed in vitro by examining the effects of either a condensed enological tannin or an hydrolyzable enological tannin on two physicochemical properties of the protein fraction of saliva, namely, its mode of diffusion on cellulose membranes and its precipitation. Comparative assays mimicking the degree of dilution experienced by saliva during a tasting assay were performed at pH 3.5 and pH 7.0. Results indicated that both enological tannins were perceived as clearly more astringent at pH 3.5 compared with pH 7.0. In addition, the effects of tannins on protein diffusion and protein precipitation were markedly exacerbated at pH 3.5.