Mixed gels of fine-stranded and particulate networks of gelatin and whey proteins

Mixed gels of gelatin and whey protein concentrate were investigated, as well as their pure systems, by tensile tests and by dynamic oscillatory measurements. The systems were studied for homogeneous particulate whey protein gels at pH 5.4 and for inhomogeneous particulate whey protein gels at pH 4.6. The influence on the systems of the Bloom number of the gelatin component has also been investigated. Results of the fracture properties, such as stress and strain at fracture, indicate a transition in rheological properties. Results of the elastic modulus, obtained by tensile measurements, as well as the storage modulus, obtained by dynamic oscillatory measurements, both agree with predictions for phase inversions from the Takayanagi models as modified by Clark, which are in agreement with the fracture properties. The transition points are different for the different mixed gel series but take place between 1 and 3 wt% gelatin and 8 wt% whey protein concentrate, depending on factors such as the microstructure of the whey protein concentrate. Dynamic oscillatory measurements showed that gel formation of whey protein concentrate is unaffected by the presence of gelatin, which is in agreement with light microscopy results. Light microscopy revealed that the mixed gel systems were bicontinuous and that the whey protein network structure was unaffected by the presence of gelatin. It is postulated that the predicted phase inversions of the mixed gels are due to a shift in rheological properties without any phase inversions in the microstructure.     

Mathematical modeling of swelling in high moisture whey protein gels

Gels prepared from whey proteins can be used for controlled release of nutrients or active ingredients in food systems. The objective of this study was to characterize the water uptake by these hydrophilic gels to aid in the design of release systems. Whey protein isolate (WPI) gels (17% w/w protein) of different aspect ratios were submersed in aqueous solution at pH 7.0. Modeling of mass uptake is presented in terms of Case I (Fickian diffusion) and Case II (kinetic) models. Due to the extent of swelling, the Fickian diffusion with moving boundaries provided the most realistic reflection of the physics. An optimization routine provided the best fit values for the diffusivity. The average diffusivity for the smaller gels (with an initial radius of 6.7 mm) was 1.40 × 10⁻¹⁰ m²/s. The average diffusivity of the larger gels (with an initial radius of 8.5 mm) was 0.79 × 10⁻¹⁰ m²/s. The average diffusivities differed due to the slight variation in the composition of the gels. The model also yielded instantaneous values of the radius and sample length. The functionality of moisture uptake and total surface area was linear. The Fickian diffusion with moving boundary model can be extended to evaluate different geometries for controlled release systems.     

Effect of protein composition on the rheological properties of acid-induced whey protein gels

Protein dispersions with different ratios of α-lactalbumin to β-lactoglobulin were heat-denatured at pH 7.5 and then acidified with glucono-δ-lactone to form gels at room temperature. Heat treatment induced the formation of whey protein polymers with reactive thiol group concentrations ranging from 1 to 50 μmol/g, depending on protein composition. During acidification, the first sign of aggregation occurred when the zeta potential reached −18.2 mV. Increasing the proportion of α-lactalbumin in the polymer dispersions resulted in more turbid gels characterized by an open microstructure. Elastic and viscous moduli were reduced, while the relaxation coefficient and the stress decay rate constants were increased by raising the proportion of α-lactalbumin in the gel. After one week of storage at 5 °C, gel hardness increased by 12%. The effect of protein composition on acid-induced gelation of whey protein is discussed in relation to the availability and reactivity of thiol groups during gel formation and storage.     

Effect of protein concentration on whey protein gels obtained by a two-stage heating process

 The influence of protein concentration on the properties of gels obtained by a two-stage heating process was determined. In the first stage, whey protein dispersion (3–10%) was heated at pH 8.0, and in the second stage it was diluted to 3% protein, adjusted to pH 7.0 and heated again. Increased protein concentration in the first stage of polymerization resulted in the gels obtained in the second stage having a lower phase angle, increased storage modulus and increased hardness. Increased protein concentration also resulted in gels with an increased optical density, which suggests thathigher protein concentration leads to more and larger aggregates. Gels obtained from dispersions preheated at a higher protein concentration had higher permeability coefficient (B _gel) values. The increase in B _gel suggests that the higher protein concentration increased the size of the aggregates, which in a second stage of heating formed a gel matrix with a larger pore size. Received: 11 February 1999     

Effects of thiol reagents and ethanol on strength of whey protein gels

Chemical reagents which interfere with secondary interactions affected gel network formation and the properties of gels made from 20% whey protein isolates. Thus, p-hydroxymercuribenzoate and N-ethylmaleimide 0–16 mmol/dm³ and dithiothreitol 0–32 mmol/dm³ reduced the hardness and cohesiveness of whey protein gels apparently by interfering with formation of disulfide bonds. The addition of ethanol (0–15%) increased the hardness of gels presumably by enhancing electrostatic interactions and hydrogen bonding.     

Characterization of cold-set gels produced from heated emulsions stabilized by whey protein

This paper reports the cold gelation of preheated emulsions stabilized by whey protein, in contrast to, in previous reports, the cold gelation of emulsions formed with preheated whey protein polymers. Emulsions formed with different concentrations of whey protein isolate (WPI) and milk fat were heated at 90 °C for 30 min at low ionic strength and neutral pH. The stable preheated emulsions formed gels through acidification or the addition of CaCl₂ at room temperature. The storage modulus (G′) of the acid-induced gels increased with increasing preheat temperature, decreasing size of the emulsion droplets and increasing fat content. The adsorbed protein denatures and aggregates at the surface of the emulsion droplets during heat treatment, providing the initial step for subsequent formation of the cold-set emulsion gels, suggesting that these preheated emulsion droplets coated by whey protein constitute the structural units responsible for the three-dimensional gel network.     

Characterisation of carrageenan and whey protein gels using NMR PGSTE diffusion experiments

NMR PGSTE diffusion experiments are used to characterise the gelling behaviour of carrageenan mixtures and whey protein mixtures. The structure of developing networks can be observed as a function of the composition of the mixture (concentration of hydrocolloids, sugars and salts), the measuring temperature, and the protein denaturation, depending on the denaturation temperature and duration. Besides the fundamental aspects of these preliminary studies, these data are helpful for modelling processes sufficiently precisely for industrial production, to guarantee specific properties concerning the flow-, gelling- and syneresis/storage-behaviour and the texture and sensorial behaviour of food products, and to realise a process and quality control of thermal protein denaturation and hydrocolloid gelling processes. Therefore, further quantitative correlations between macroscopic quantities and the available NMR parameters must be established to trigger industrial applications of NMR diffusion experiments.     

Utilizing whey protein isolate and polysaccharide complexes to stabilize aerated dairy gels

Heated soluble complexes of whey protein isolate (WPI) with polysaccharides may be used to modify the properties of aerated dairy gels, which could be formulated into novel-textured high-protein desserts. The objective of this study was to determine the effect of polysaccharide charge density and concentration within a WPI-polysaccharide complex on the physical properties of aerated gels. Three polysaccharides having different degrees of charge density were chosen: low-methoxyl pectin, high-methoxyl type D pectin, and guar gum. Heated complexes were prepared by heating the mixed dispersions (8% protein, 0 to 1% polysaccharide) at pH 7. To form aerated gels, 2% glucono-δ-lactone was added to the dispersions of skim milk powder and heated complex and foam was generated by whipping with a handheld frother. The foam set into a gel as the glucono-δ-lactone acidified to a final pH of 4.5. The aerated gels were evaluated for overrun, drainage, gel strength, and viscoelastic properties. Without heated complexes, stable aerated gels could not be formed. Overrun of aerated gel decreased (up to 73%) as polysaccharide concentration increased from 0.105 to 0.315% due to increased viscosity, which limited air incorporation. A negative relationship was found between percent drainage and dispersion viscosity. However, plotting of drainage against dispersion viscosity separated by polysaccharide type revealed that drainage decreased most in samples with high-charge-density, low-methoxyl pectin followed by those with low-charge-density, high-methoxyl type D pectin. Aerated gels with guar gum (no charge) did not show improvement to stability. Rheological results showed no significant difference in gelation time among samples; therefore, stronger interactions between WPI and high-charge-density polysaccharide were likely responsible for increased stability. Stable dairy aerated gels can be created from WPI-polysaccharide complexes. High-charge-density polysaccharides, at concentrations that provide adequate viscosity, are needed to achieve stability while also maintaining dispersion overrun capabilities.     

Effect of monoglycerides and diglycerol-esters on viscoelasticity of heat-set whey protein emulsion gels

Summary Effects of small-molecule surfactants (emulsifiers) on the small-deformation viscoelastic properties of heat-set whey protein emulsion gels have been investigated using a controlled stress rheometer. The surfactants used in this investigation were the water-soluble diglycerol monolaurate (DGML) and diglycerol monooleate (DGMO), and the oil-soluble glycerol monooleate (GMO). The elastic modulus of the emulsion gel was found to decrease in the presence of a small amount of surfactant, but then to recover at higher surfactant concentrations. The initial reduction in modulus correlates with protein displacement from the oil droplet surface. The recovery of the storage and loss moduli at higher surfactant concentrations of DGML or DGMO may be due to the depletion flocculation of the emulsion prior to heat-treatment. However, for systems containing high content of GMO in the oil phase, the recovery of the moduli is probably owing mainly to the smaller average particle size. Effects of surface monolayer composition, droplet aggregation and average particle size were discussed. The behaviour obtained here was compared with results for previously investigated whey protein emulsion gel systems containing different emulsifiers.     

Effect of whey protein enzymatic hydrolysis on the rheological properties of acid-induced gels

A protein dispersion blend of β-lactoglobulin and α-lactalbumin was heat-denatured at pH 7.5, hydrolyzed by α-chymotrypsin and then acidified with glucono-δ-lactone to form gels at room temperature. Heat treatment induced the formation of whey protein polymers with high concentration of reactive thiol groups (37 μmol/g). The reactive thiol group concentration was reduced by half after 40 min enzymatic hydrolysis. It was further reduced after enzyme thermal deactivation. During acidification, the first sign of aggregation for hydrolyzed polymers occurred earlier than for non hydrolyzed polymers. Increasing the hydrolysis duration up to 30 min resulted in more turbid gels characterized by an open microstructure. Elastic and viscous moduli were both reduced, while the relaxation coefficient and the stress decay rate constants were increased by increasing the hydrolysis duration. After one week storage at 5 °C, the hardness of gels made from hydrolyzed polymers increased by more than 50%. The effect of polymer hydrolysis on acid-induced gelation is discussed in relation to the availability and reactivity of thiol groups during gel formation and storage.     

Effect of denatured whey protein concentrate and its fractions on rennet-induced milk gels

Denatured whey protein concentrate was fractionated by centrifugation to study the effect of its different components (sedimentable aggregates, non-sedimentable component, and diffusible component) on rennet-induced coagulation of milk and gel contraction capacity. Milk coagulation properties were characterized by optical density measurement and dynamic rheometry. The contraction kinetics of the gel during cooking was also characterized. The diffusible component of denatured whey protein concentrate showed no significant effect on coagulation or contraction parameters. Sedimentable aggregates negatively influenced the kinetics of rennet gel formation, as measured by rheology; these aggregates also reduced the contraction capacity of the gel. The non-sedimentable component negatively influenced milk coagulation properties, as measured with both optical and rheological methods, and decreased the contraction capacity of the gel. The results suggest that, beyond the effect of sedimentable whey protein aggregates, soluble proteinaceous complexes (non-sedimentable and non-diffusible) could interact with renneted casein micelles and limit gel formation and contraction.     

Kinetics of chemical marker formation in whey protein gels for studying microwave sterilization

The kinetics of 4-hydroxy-5-methy-3(2H)-furanone (M-2) formation in a model food system (20% whey protein gel) was determined for studying cumulative time–temperature effects in high-temperature-short-time processes. M-2 was formed from -ribose and amines through non-enzymatic browning reactions and enolization under low acid conditions (pH > 5). The order of the reaction for M-2 formation was determined by non-linear regression analysis and further confirmed by graphical method. M-2 formation followed a first-order kinetics and the rate constant temperature dependence was described using an Arrhenius relationship. The reaction rates and activation energy were determined using two-step, multi-linear and non-linear regression analyses. This study also demonstrated the use of M-2 formation in determining the cumulative heating effect in a model food system subjected to 915 MHz microwave heating.     

The breakdown properties of heat-set whey protein emulsion gels in the human mouth

Differently structured whey protein emulsion gels were formed by heating at different concentrations of NaCl. The formation of gels was monitored by oscillatory rheometry. The large deformation properties relevant to breakdown properties in the human mouth were measured by a uniaxial compression test and fracture wedge set test using a texture analyzer. A panel of 8 subjects was used to examine the in-mouth behaviours of gels including mastication parameters, degree of fragmentation and oil droplet release. The results showed that in general the gel hardness increased with increasing NaCl concentration. The gels containing 10/25 and 100/200 mM NaCl were characterized as being soft and hard, respectively. These soft and hard gels had different breakdown patterns in the mouth. On the other hand, sensory experiments showed the gel with 10 mM NaCl needed a significantly lower number of chewing cycles (19.4 ± 2.1) compared with gels with higher NaCl. The values of median size of particles in masticated gels containing 10, 25, 100 and 200 mM NaCl were about 4.00, 2.85, 1.05 and 0.95 mm, respectively, which suggested that higher hardness led to greater fragmentation in the human mouth. The fragmentation of the gel was highly correlated with functions of the mechanical properties. There was no obvious coalescence of the oil droplets during oral processing and only very few oil droplets were released from protein matrix during mastication.     

Microstructural features of composite whey protein/polysaccharide gels characterized at different length scales

Mixed biopolymer gels are often used to model semi-solid food products. Understanding of their functional properties requires knowledge about structural elements composing these systems at various length scales. This study has been focused on investigating the structural features of mixed cold-set gels consisting of whey protein isolate and different polysaccharides at different length scales by using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Whey protein cold-set gels were prepared at different concentrations to emulate stiffness of various semi-solid foods. Mixed gels contained different concentrations of gellan gum, high methyl pectin or locust bean gum. Results obtained with CLSM, at the micrometer length scale, indicated the homogeneous nature of the investigated gels. Results obtained with SEM, at the sub-micron length scale, indicated the presence of spherical protein aggregates. During the gel preparation (acidification), the presence of polysaccharides in the whey protein gels led to on initially segragative phase separation into a gelled protein phase and a polysaccharide/serum phase at a micrometer length scale. At the final pH of the gels (pH 4.8, i.e. below the pI of whey proteins), the negatively charged polysaccharides interacted with the protein phase and their spatial distribution was effected by charge density. Polysaccharides with a higher charge density were more homogeneously distributed within the protein phase. Neutral polysaccharide, locust bean gum, did not interact with the protein aggregates but was present in the serum phase. Using SEM, a new type of microstructure formed in the whey protein/polysaccharide gels was characterized. It composed of a protein continuous, porous network at the length scale of 100 μm, coexisting next to the pools of serum which contained spherical protein-rich domains. Heterogeneity of the structure strongly related to the macroscopic behavior of the gels under large deformation. Upon uniaxial compression these heterogeneous gels releases a large amount of serum. Combination of the results of two microscopic techniques, CLSM and SEM, appeared to offer unique possibilities to characterize the structural elements of whey protein/polysaccharide cold-set gels over a wide range of length scales.     

Polyelectrolyte screening effects on the dissolution of whey protein gels at high pH conditions

The literature reports an optimum NaOH concentration for the alkaline cleaning of whey deposits or gels; at NaOH concentrations higher than this optimum, cleaning proceeds much more slowly. Although this phenomenon is of great importance in the cleaning of dairy equipment, no conclusive physical explanation has yet been presented. In this study, we present strong evidence that the dissolution rate is affected by the equilibrium-swelling ratio in β-lactoglobulin (βLg) gels. The swelling ratio is greatly reduced in the presence of salts due to the polyelectrolyte screening effect of the cations. This has been observed in free-swelling βLg gels using gravimetrical analysis and in the uniaxial swelling of WPC gel deposits using fluid dynamic gauging. At high dissolution pH (>13.3), the high Na⁺ concentration reduces swelling in spite of the high surface charge of the protein. It is proposed that the reduction of the free volume inside the gel impedes the transport of the protein aggregates out of the NaOH penetration zone. We have also observed that the final dissolution rate of gels pre-soaked in 1 M NaOH or NaCl is similar, despite the difference in pH, and much lower than for untreated gels: the high Na⁺ concentration in the soaked gels hinders swelling, inhibiting the disentanglement of the protein clusters regardless of the high pH.     

Combined microscopic and dynamic rheological methods for studying the structural breakdown properties of whey protein gels and emulsion filled gels

The microstructural and large deformation rheological properties of model food gels were studied by performing notch propagation tensile testing on the gels using a tensile stage and observing changes in the microstructure of the gels during tensile testing using confocal laser scanning microscopy (CLSM). Heat-set whey protein (WP) gels containing either added sodium caseinate (NaCN) or sunflower oil droplets emulsified with WP or NaCN as the emulsifier protein were prepared in 0 or 50 mM NaCl. The WP gel structure strengthened in the presence of added NaCl and NaCN. The rheological properties of WP gels containing sunflower oil droplets emulsified with WP or NaCN were influenced by the NaCl concentration, oil concentration and extent of oil droplet aggregation in the gel or by the type of emulsifier protein used. During tensile testing, the notch length in all gels increased above a certain critical stress, leading to fracture of the gels through the notch. Also, the microstructural changes in the oil phase of emulsion filled gels subjected to tensile testing were influenced by the structural properties of the WP gel matrix and the proximity of the oil droplet to the fracture path.     

Cold‐set whey protein gels induced by calcium or sodium salt addition

Cold‐set whey protein isolate (WPI) gels formed by sodium or calcium chloride diffusion through dialysis membranes were evaluated by mechanical properties, water‐holding capacity and microscopy. The increase of WPI concentration led to a decrease of porosity of the gels and to an increase of hardness, elasticity and water‐holding capacity for both systems (CaCl₂ and NaCl). WPI gels formed by calcium chloride addition were harder, more elastic and opaque, but less deformable and with decreased ability to hold water in relation to sodium gels. The non linear part of stress–strain data was evaluated by the Blatz, Sharda, and Tschoegl equation and cold‐set gels induced by calcium and sodium chloride addition showed strain‐weakening and strain‐hardening behaviour, respectively. The fractal structure of the gels indicated a weak‐link behaviour. For WPI gels results suggest intrafloc links, formed at heating step, which were more rigid than the interfloc links, promoted by salt addition.     

Effect of heating temperature,pH, concentration and starch/whey protein ratio on the viscoelastic properties of corn starch/whey protein mixed gels

The viscoelastic properties of corn starch (CS) gels were more dependent on heating temperature, while the properties of whey protein isolate (WPI) gels were more dependent on pH. Thus heating temperature (75, 85, 95 °C) and pH (5, 7, 9) were varied to obtain a series of mixed gels with interesting viscoelastic properties. WPI gels showed extensive stress relaxation (SR) indicative of a highly transient network structure, while CS gels relaxed very little in 2000 s. Based on SR results, it appeared that CS/WPI mixed gels with 25 and 50% CS formed compatible network structures at 15% total solids only at pH 9. This supposition was supported by SEM microstructures obtained for dehydrated gels and a synergistic increase in the large‐strain fracture stress for these gels. Some synergy was also found for mixed gels at 30% total solids at pH 9, while at pH 7 the mixed gels seemed to contain separate additive WPI and CS networks unlike the case for pH 7 at 15% total solids. In both cases (15 and 30% total solids) the degree of elasticity of the mixed gels decreased as the WPI content increased. Mixed gels (CS:WPI = 0.5) at pH 9 showed increased fracture stress and fracture strain relative to the same gels at pH 7. This suggests that a unique chemical compatibility exists at pH 9 and results in gels that combine the elasticity of CS and the internal stress dissipation of WPI. © 2001 Society of Chemical Industry