Shear-induced structuring of particulate whey protein gels

The effects of steady shear on particulate whey protein isolate (WPI) gels, at pH 5.4, have been investigated by light microscopy (LM) and dynamic oscillatory measurements. The steady shear was performed on suspensions at constant rates between 0.5 and 126/s. The gel point under static conditions (T_g) was around 78 °C and the shearing was performed during heating from 20 to 76 or to 82 °C. The gel point was postponed by the shear up to 82 °C. Steady shear up to 76 °C, at rates less than 6/s, resulted in a weaker storage modulus (G′), less frequency dependence and a higher stress at fracture compared to the unsheared gel. Steady shear up to 82 °C, at rates below 6/s, resulted in the formation of two different types of network structure. One structure was similar in appearance to the unsheared network, showing pores in the range of 50 μm. The other structure was dense, composed of smaller particles than the unsheared network and with pores in the range of 10 μm. The gels composed of two structures showed a lower G′ and stress at fracture compared to the unsheared gel. A shear rate above 24/s up to 76 °C resulted in irregular networks, which were composed of two different types of structures. One was loose and open, similar in appearance to the unsheared network structure. The other structure was dense and compact, and was present as individual aggregates. These gels also showed a weaker G′ than the unsheared gel. A shearing up to 82 °C at rates above 24/s resulted in a coarse, inhomogeneous network structure. The gels showed a weak G′, indicating aggregate break-up during the steady shear.     

Influence of Shear Treatments on Consistency and Gelling Properties of Whey Protein Isolate Suspensions

Suspensions of whey protein isolate (WPI) subjected to controlled shearing by capillary extrusion, and Theologically tested after shearing, showed increasing consistency index as applied shear rate increased. Sheared suspensions gelled more rapidly than unsheared ones and produced stronger gels. SEM micrographs showed the development of a fine fibrous-like structure in the sheared suspensions, indicating numerous sites of protein-protein interaction. Sheared WPI suspensions, added as a replacement for fat in frankfurter formulations, showed excellent water-holding capacity, producing low-fat frankfurters with elastic moduli in the same order of magnitude as commercial frankfurters.     

Study on the rheological properties of heat-induced whey protein isolate-dextran conjugate gel

Effect of glycosylation on the rheological properties of whey protein isolate (WPI) during the heat-induced gelation process was evaluated. Significant changes in browning intensity, free amino groups content and SDS-PAGE profile showed that the conjugate of WPI and dextran (150 kDa) was successfully prepared using the traditional dry-heating treatment. For the conjugate, during the heating and cooling cycle, the curves of G′ and G″ were considerably shifted to lower values and their shapes varied comparing to the corresponding spectra of initial WPI and WPI + dextran mixture. After holding at 25 °C, G' reached a value of about 2200 Pa, only a tenth of the value that obtained in the initial WPI gel. Moreover, frequency sweep measurements revealed that the stiffness of gel was greatly reduced in the conjugate, although a typical elastic gel was still formed. All data showed that the rheological properties of thermal gelation could be modified upon the covalent attachment of dextran.     

Thermodynamic incompatibility between denatured whey protein and konjac glucomannan

The current study investigated the effect of a neutral polysaccharide, konjac glucomannan, on the heat-induced gelation of whey protein isolate (WPI) at pH 7. Oscillatory rheology (1 rad/s; 0.5% strain), differential scanning calorimetry and confocal laser scanning microscopy were used to investigate the effect of addition of konjac in the range 0-0.5% w/w, on the thermal gelation properties of WPI. The minimum gelling concentration for WPI samples was 11% w/w; the concentration was therefore held constant at this value. Gelation of WPI was induced by heating the samples from 20 to 80 °C, holding at 80 °C for 30 min, cooling to 20 °C, and holding at 20 °C for a further 30 min. On incorporation of increasing concentrations of konjac the gelation time decreased, while the storage modulus (G′) of the mixed gel systems increased to ∼1450 Pa for 11% w/w WPI containing 0.5% w/w konjac gels, compared to 15 Pa for 11% w/w WPI gels (no konjac). This increase in gel strength was attributed to segregative interactions between denatured whey proteins and konjac glucomannan.     

Physical properties of acid milk gels prepared at 37°C up to gelation but at different incubation temperatures for the remainder of fermentation

We investigated the effect of altering temperature immediately after gels were formed at 37°C. We defined instrumentally measurable gelation (IMG) as the point at which gels had a storage modulus (G′) ≥5 Pa. Gels were made at constant incubation temperature (IT) of 37°C up to IMG, and then cooled to 30 or 33.5, or heated to 40.5 or 44°C, at a rate of 1°C/min and maintained at those temperatures until pH 4.6. Control gel was made at 37°C (i.e., no temperature change during gelation/gel development). Gel formation was monitored using small strain dynamic oscillatory rheology, and the resulting structure and physical properties at pH 4.6 were studied by fluorescence microscopy, large deformation rheology, whey separation (WS), and permeability (B). A single strain of Streptococcus thermophilus was used to avoid variations in the ratios of strains that could have resulted from changes in temperature during fermentation. Total time required to reach pH 4.6 was similar for samples made at constant IT of 37°C or by cooling after IMG from 37 to either 30 or 33.5°C, but gels heated to 40 or 44°C needed less time to reach pH 4.6. Cooling gels after IMG resulted in an increase in G′ values at pH 4.6, a decrease in LT_max, WS, and B, and an increase in the area of protein aggregates of micrographs compared with the control gel made at constant IT of 37°C. Heating gels after IMG resulted in a decrease in G′ values at pH 4.6 and an increase in LT_max values and WS. The physical properties of acid milk gels were dominated by the temperature profile during the gel-strengthening phase that occurs after IMG. This study indicates that the final properties of yogurt greatly depend on the environmental conditions (e.g., temperature, time/rate of pH change) experienced by the casein particles/clusters during the critical early gel development phase when bonding between and within particles is still labile. Cooling of gels may encourage inter-cluster strand formation, whereas heating of gels may promote intra-cluster fusion and the breakage of strands between clusters.     

Influence of shearing on the physical characteristics and rheological behaviour of an aqueous whey protein isolate–κappa-carrageenan mixture

The incompatibility of whey protein isolate (WPI) and κappa-carrageenan (κ-car) in aqueous mixtures has been extensively studied under quiescent conditions; however, the effect of shear on segregative phase separation is still not fully understood. The present work reports for the first time quantitatively the effect of shearing on the segregative phase separation behaviour of these two polymers. Demixing was observed at pH 7.0 and 22 °C, determining the phase diagram and rheological properties of the mixtures. Phase diagrams were derived after heating and cooling mixes at a constant shear rate (28 s⁻¹). The phase behaviour was compared to that of the same mixtures under quiescent conditions. The shearing process affects segregative phase separation, causing a shift in the position of the binodal towards lower concentrations of WPI. The bottom layer contained a higher ratio of WPI while the upper layer was enriched in κ-car. The addition of κ-car to WPI solutions led to a much stiffer heat-induced gel than that prepared with WPI heated in isolation. The height of the plateau of the final elastic modulus G′_∞(t) depends on the position of the system on the phase diagram. Using a selected tie line, the viscosity of different systems measured at 80 °C was more influenced by the amount of WPI, than by the κ-car concentration. Shear treatment of segregative phase separating systems offer a way to modulate the functional properties of the ingredients and the texture of the final product.     

The effect of shear on the rheology and structure of heat‐induced whey protein gels

The influence of mechanical shearing on the small deformation properties and microstructure of heat‐induced whey protein gel has been studied. The viscoelastic properties of these gels at different concentrations of 10% and 20% (w/w) exposed to different shear rates of 0, 50, 100, 200 and 500 s^?1 during gelation were measured using dynamic oscillatory rheometry. The structure of both the shear treated and unsheared gels was then investigated using light microscopy. The results showed that the storage modulus of the gels at both concentrations was increased by increasing the shear rate exposure during gelation while the shear‐treated gels were more elastic and showed frequency‐independent behaviour. As the total protein concentration of the gel increased, the viscoelastic properties of the gels also increased significantly and the gels showed greater elasticity. The gels obtained from the higher shear rate exposure were stronger with higher elastic moduli at both protein concentrations. Images of the gels obtained using light microscopy showed that shearing resulted in phase separation and some aggregation in the structure of the gels at both concentrations. However, the shearing rates applied in this study were not enough to cause aggregation breakdown in the gel network.     

Gelation of whey protein and xanthan mixture: Effect of heating rate on rheological properties

The effects of heating rate and xanthan addition on the gelation of a 15% w/w whey protein solution at pH 7 and in 0.1 M phosphate buffer were studied using small-amplitude oscillatory shear (SAOS) rheological measurements and uniaxial compression tests. WPI solutions were heated from 25 to 90 °C at five heating rates (0.1, 1, 5, 10 and 20 °C/min). Gelation temperature of WPI decreased with decreasing of heating rates and with xanthan addition. Under uniaxial compression, the WPI gels prepared with no more than 0.2% w/w xanthan exhibited distinct fracture point and were tougher (i.e. higher fracture stress and fracture strain) than the gels prepared with no less than 0.5% w/w xanthan. In general, the fracture strain of WPI gels increased with heating rate, though not significantly, at all xanthan contents investigated. However, the fracture stress of WPI gels, generally, decreased with heating rate when xanthan content was 0–0.2% and increased with heating rate when xanthan content was 0.5 and 1%.     

Rheological properties of rennet casein-whey protein gels prepared at different mixing speeds

The rheological properties at small (oscillatory shear) and large (uniaxial compression) deformations of heat-induced gels (80 °C for 20 min, pH 7.3) containing 25% rennet casein (RCN), 2.5% disodium phosphate and 0%, 2.3% or 6.3% of whey protein isolate (WPI) were measured for samples cooked in a torque-rheometer at mixing speeds within a range of 20–200 rpm (shear rates: ∼15–230 s⁻¹). In addition, microstructure analyses were performed, separately staining RCN and WPI, by Confocal Scanning Laser Microscopy (CSLM). Both small and large deformation tests indicated that increasing addition of WPI prior to the cooking process of RCN resulted in gels exhibiting higher storage and deformability moduli than WPI-free samples. Increasing shear rates during cooking also affected the rheological properties of RCN–WPI gels, and stronger gels were formed as the shear rate during cooking was increased. Despite the data dispersion among replicates, the effect of shear rate on gel strength were evident for RCN gels with 6.3% WPI and relatively clear for gels with 2.3% WPI; however, the trend was uncertain for WPI-free RCN gels. Possible explanations for this observation are that when increasing WPI levels in the presence of RCN and heat, disulfide-thiol exchange reactions between denatured WPI and κ-casein (κ-CN) are increased and possibly promoted by shear rate, resulting in stronger and more cross-linked gel structure. CSLM results were not conclusive to support this hypothesis.     

Modulation of rheological properties by heat-induced aggregation of whey protein solution

Heat-induced protein aggregation at low protein concentrations generally leads to higher viscosities. We here report that aggregated protein can yield weaker gels than those from native protein at the same concentration. Aggregated protein was produced by heating a solution of whey protein isolate (WPI) at 3% and 9% w/w. The higher protein concentration resulted in a larger aggregate size and a higher intrinsic viscosity. The protein fraction in native WPI had the smallest size and the lowest intrinsic viscosity. The same trend was observed for the shear viscosity after concentrating the suspensions containing aggregates to around 15% w/w. Suspensions containing aggregates that were produced from a higher concentration possessed a higher viscosity. After reheating the concentrated suspensions, the suspension from the 9% w/w aggregate system produced the weakest gel, followed by the one from 3% w/w, while the native WPI yielded the strongest gel. Reactivity of the aggregates was also an important factor that influenced the resulting gel properties. We conclude that aggregation of whey protein solution is a feasible route to manipulate the gel strength of concentrated protein systems, without having to alter the concentration of the protein.     

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.     

Modification of rheological properties of whey protein isolates by limited proteolysis

Whey protein isolate (WPI) was subjected to limited tryptic hydrolysis and the effect of the limited hydrolysis on the rheological properties of WPI was examined and compared with those of untreated WPI. At 10% concentration (w/v in 50 mM TES buffer, pH 7.0, containing 50 mM NaCl), both WPI and the enzyme-treated WPI (EWPI) formed heat-induced viscoelastic gels. However, EWPI formed weaker gels (lower storage modulus) than WPI gels. Moreover, a lower gelation point (77 °C) was obtained for EWPI as compared with that of WPI which gelled at 80 °C only after holding 1.4 min. Thermal analysis and aggregation studies indicated that limited proteolysis resulted in changes in the denaturation and aggregation properties. As a consequenece, EWPI formed particulated gels, while WPI formed fine-stranded gels. In keeping with the formation of a particulate gel, Texture Profile Analysis (TPA) of the heat-induced gels (at 80 °C for 30 min) revealed that EWPI gels possessed significantly higher (p < 0.05) cohesiveness, hardness, gumminess, and chewiness but did not fracture at 75% deformation. The results suggest that the domain peptides, especially β-lactoglobulin domains released by the limited proteolysis, were responsible for the altered gelation properties.     

Impact of Residual Lactose on Dry Heat-Induced Pre-texturization of Whey Proteins

Controlling denaturation/aggregation of whey proteins during their pre-texturization is highly critical to avoid variability in their functional properties. We investigated how the dry heat-induced (16 h at 100 °C and a_w?=?0.23) pre-texturization of whey protein isolate (WPI) is affected by traces of remaining lactose (0.3–2.0%) and how it influences its subsequent gelling properties. Lactose even in trace quantities developed intense browning of WPI. Dry-heating conditions used in this study mainly developed soluble aggregates stabilized by covalent crosslinks other than disulfide bonds. The extent of aggregation and size of aggregates were drastically increased with increasing lactose. Intermediate quantity (39–46%) of soluble aggregates improved the gel strength, while excessive aggregation (>?50%) resulted in loss of gel strength. Elasticity of gels was also increased by increasing protein aggregates. This study suggests that the traces of lactose that remain in WPI are critical for controlling its pre-texturization by dry heating and its subsequent gelling properties.     

Factors Influencing Whey Protein Gel Rheology: Dialysis and Calcium Chelation

Influence of dialyzable compounds on the Theological properties (shear stress and shear strain at failure) of heat-induced whey protein concentrate (WPC) and whey protein isolate (WPI) gels was examined. Dialyzing WPC and WPI suspensions prior to gelation increased the stress of two of three WPC gels and a WPI gel. Dialysis also significantly increased the strain of the same two WPC gels, normalizing all strain values. Replacement of calcium lost through dialysis did not significantly change gel rheology. However, chelating calcium caused a significant decrease in the stress of all gels: a minimum amount of calcium and/or a calcium complex appears to have a major role in whey protein gelation.     

Effect of non-starch polysaccharides on the in vitro digestibility and rheological properties of rice starch gel

The starch digestibility and rheological properties of gels were evaluated in the presence of three non-starch polysaccharides (agar, xanthan gum and konjac glucomannan) with rice starch. Each polysaccharide was added to 30% (w/w) rice starch suspension at defined concentrations and starch gels were prepared. The extent of starch gel digestibility was determined by an in vitro method and rheological properties by a dynamic oscillatory test and a compression test. The added polysaccharides suppressed starch hydrolysis in the gels compared with the control, and a concentration dependency of this suppressive effect was observed. Adding agar and xanthan gum increased the storage shear modulus (G′) of starch gels, while adding konjac glucomannan decreased G′ values. The results indicate that the suppressive effect of non-starch polysaccharides on starch digestibility appears to be not only due to the rigidity of the gel, but also the interaction between starch and non-starch polysaccharides.     

Solubilized micellar calcium induced low methoxyl-pectin aggregation during milk acidification

Low methoxyl (LM) pectin was combined with 3-kDa molecular weight cut-off permeates from milk subjected to pH 6.7 to 5 and 7°C or 40°C with the objective of studying the effect of solubilized micellar calcium on viscoelastic properties of LM-pectin-milk mixes. Lowering the pH of skim milk with hydrochloric acid during ultrafiltration gradually promoted permeates to exhibit gel-like behavior when combined with LM-pectin. The onset of the gel-like behavior (G′ > 1) occurred at a higher pH when permeates were obtained from milk filtered at 7°C compared with 40°C. As pH value during ultrafiltration approached 5 and regardless of temperature, G′ for permeate-pectin mixes approached the same values (∼70 Pa) as G′ for skim milk-pectin mixes. In all cases G′ was highly correlated with free calcium concentration (r > 0.95). The gradual acidification of skim milk-LM-pectin using glucono-δ-lactone, promoted a sharp increase in storage modulus as pH approached 5.2 and a maximum G′ increment (ΔG′) at pH ∼4.9. From pH 4.9 to 4, G′ continued to increase but at smaller increments. It was concluded that LM-pectin-casein micelle interaction in milk is a 2-step process: 1) solubilized micellar calcium dependent pectin-pectin interaction as pH approaches 5.0 to 4.9, and 2) pectin-casein micelle interaction in the 5.0-4.9 to 4.0 pH range.     

Whey protein aggregate formation during heating in the presence of κ-carrageenan

The effect of a negatively charged polymer, κ-carrageenan, on the aggregation behaviour of whey proteins during heating was studied. Aqueous solutions of whey protein isolate (WPI) at 0.5% were heated in the presence of κ-carrageenan (0.1%) at pH 7.0. This concentration was chosen as optimal in the detection of the intermediate aggregates during chromatographic analysis. The residual unaggregated protein, the intermediate aggregates and the soluble aggregates were all examined as a function of heating time and temperature, using size-exclusion chromatography coupled with light scattering detection. The presence of κ-carrageenan did not affect the aggregation of whey proteins heated at 75 °C; however, a change in the mechanism of aggregation seemed to occur at higher temperatures, and intermediates with higher molecular mass formed at 85 °C. At 90 °C, in the presence of κ-carrageenan, the extent of WPI aggregation was much larger, as soluble aggregates were no longer present and less residual protein was recovered in the unaggregated peak.     

Xanthan Enhances Water Binding and Gel Formation of Transglutaminase-Treated Porcine Myofibrillar Proteins

ABSTRACT: In this study, the effect of xanthan on dynamic rheological properties, textural profile, and water binding of transglutaminase (TG)-treated myofibrillar protein (MP) gels was investigated. In experiment 1, MP suspensions (40 mg/mL protein, 0.6 M NaCl) at pH 6.45 with or without 0.05% xanthan were treated with 0%, 0.1%, 0.2%, 0.3%, 0.4%, and 0.5% TG; in experiment 2, MP suspensions (40 mg/mL protein, 0.6 M NaCl) at pHs 6.13, 6.30, 6.45, 6.69 with or without 0.05% xanthan were treated with 0.3% TG. Treated samples were analyzed with differential scanning calorimetry for thermal stability and oscillatory rheometry and Instron penetration tests for gelation properties. The TG treatments lowered the transition temperature (T_m) of MP by as much as 6 °C (P < 0.05) but increased apparent enthalpy of denaturation. However, there was no detectable thermal stability difference between MP samples with or without xanthan. The shear storage modulus (G′) of MP gels increased markedly upon treatments with ≥0.3% TG, and the presence of xanthan further enhanced the gel strength (P < 0.05). The addition of 0.05% xanthan decreased cooking loss of TG-treated MP gels by 17% to 23% when compared with gels without xanthan at all pH levels evaluated (6.13 to 6.67). Thus, the combination of TG and xanthan offered a feasible means to promote cross-linking and gelation of MP while reducing cooking losses.     

Comparative effect of thermal treatment on the physicochemical properties of whey and egg white protein foams

The optimization of the functionalities of commercial protein ingredients still constitutes a key objective of the food industry. Our aim was therefore to compare the effect of thermal treatments applied in typical industrial conditions on the foaming properties of whey protein isolate (WPI) and egg white proteins (EWP): EWP was pasteurized in dry state from 1 to 5 days and from 60 °C to 80 °C, while WPI was heat-treated between 80 °C and 100 °C under dynamic conditions using a tubular heat exchanger. Typical protein concentrations of the food industry were also used, 2% (w/v) WPI and 10% (w/v) EWP at pH 7, which provided solutions of similar viscosity. Consequently, WPI exhibited a higher foamability than EWP. For WPI, heat treatment induced a slight decrease of overrun when temperature was above 90 °C, i.e. when aggregation reduced too considerably the amount of monomers that played the key role on foam formation; conversely, it increased foamability for EWP due to the lower aggregation degree resulting from dry heating compared to heat-treated WPI solutions. As expected, thermal treatments improved significantly the stability of WPI and EWP foams, but stability always passed through a maximum as a function of the intensity of heat treatment. In both cases, optimum conditions for foam stability that did not impair foamability corresponded to about 20% soluble protein aggregates. A key discrepancy was finally that the dry heat treatment of EWP provided softer foams, despite more rigid than the WPI-based foams, whereas dynamically heat-treated WPI gave firmer foams than native proteins.     

Reducing the stiffness of concentrated whey protein isolate (WPI) gels by using WPI microparticles

Concentrated protein gels were prepared using native whey protein isolate (WPI) and WPI based microparticles. WPI microparticles were produced by making gel pieces from a concentrated WPI suspension (40% w/w), which were dried and milled. The protein within the microparticles was denatured and the protein concentration after drying was similar to the native WPI powder. WPI microparticles had irregular shape with an average size of about 70 μm. They absorbed water when dispersed in water, but the dispersion did not gel upon heating. Replacing part of the native WPI powder with WPI microparticles in the protein gel resulted in lower gel stiffness compared with a gel with the same overall protein concentration but without microparticles. However, microparticles also strengthened the continuous phase because they take up water from this phase. This might increase gel stiffness more than would be expected from an inert particle/filler. There was also good bonding between the microparticles and the WPI continuous phase in the gel, which contributed to gel stiffness.