Rheological properties of rennet gels containing milk protein concentrates

Different milk protein concentrates (MPC), with protein concentrations of 56, 70, and 90%, were dispersed in water under different treatments (hydration, shear, heat, and overnight storage at 4°C), as well as in a combination of all the treatments in a factorial design. The particle size distribution of the dispersions was then measured to determine the optimal conditions for the dispersion. Heating at 60°C for 30 min with 5 min of shear was chosen as the best condition to dissolve MPC powders. The samples were also characterized for composition, presence of protein aggregates, and ratio of calcium to protein. The total calcium present in MPC increased with increasing concentration of protein; however, the total calcium-to-protein ratio was lower in MPC90 than in MPC56 and MPC70. The level of whey protein denaturation, the presence of κ-casein-whey protein aggregates in the supernatant after centrifugation, and the amount of caseins dissociated from the micelle increased as the protein concentration in the powder increased. The total amount of casein macropeptide released was lower in samples from powders with a higher protein concentration than for MPC56 or the skim milk control. The gelation behavior of reconstituted MPC was tested in systems dispersed in water (5% protein) as well as in systems dispersed in skim milk (6% protein). The gelation time of MPC dispersions was considerably lower and the gel modulus was higher than those of reconstituted skim milk with the same protein concentration. When MPC dispersions were dialyzed against skim milk, a significant decrease in the gelation time and modulus were shown, with a complete loss of gelling functionality in MPC90 dispersed in water. This demonstrated that the ionic equilibrium was key to the functionality of MPC.     

Rheological and structural characterization of gels from whey protein hydrolysates/locust bean gum mixed systems

The gelling ability of whey proteins can be changed by limited hydrolysis and by the addition of other components such as polysaccharides. In this work the effect of the concentration of locust bean gum (LBG) on the heat-set gelation of aqueous whey protein hydrolysates (10% w/w) from pepsin and trypsin was assessed at pH 7.0. Whey protein concentrate (WPC) mild hydrolysis (up to 2.5% in the case of pepsin and 1.0% in the case of trypsin) ameliorates the gelling ability. The WPC synergism with LBG is affected by the protein hydrolysis. For a WPC concentration of 10% (w/w), no maximum value was found in the G′ dependence on LBG content in the case of the hydrolysates, unlike the intact WPC. However, for higher protein concentrations, the behaviour of gels from whey proteins or whey protein hydrolysates towards the presence of LBG becomes very similar. In this case, a small amount of LBG in the presence of salt leads to a big enhancement in the gel strength. Further increases in the LBG concentration led to a decrease in the gel strength.     

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.     

Rheological properties of reduced-fat and low-fat ice cream containing whey protein isolate and inulin

Instrumental analyses were used to evaluate the rheological properties of regular (10%), reduced-fat (6%) and low-fat (3%) ice cream mixes and frozen ice creams stored at −18 °C. The reduced-fat and low-fat ice creams were prepared using 4% whey protein isolate (WPI) or 4% inulin as the fat replacement ingredient. The composition, colour, apparent viscosity, consistency coefficient, flow behaviour index, hardness and melting characteristics were measured. No effect of WPI or inulin was obtained on the colour values. Compared with regular ice cream, WPI changed rheological properties, resulting in significantly higher apparent viscosities, consistency indices and greater deviations from Newtonian flow. In addition, both hardness and melting resistance significantly increased by using WPI in reduced-fat and low-fat ice creams. Inulin also increased the hardness in comparison to regular ice cream, but the products made with inulin melted significantly faster than the other samples.     

Gelation of casein-whey protein mixtures

Heated milk consists of a mixture of whey protein-coated casein micelles and soluble whey protein aggregates. The acid-induced gelation properties of heated milk are consistently different from those of unheated milk—i.e., a shift in gelation pH, stronger gels, and a different microstructure of the gels. In this study we investigated the role of the different fractions of denatured whey proteins on the acid-induced gelation, the gel hardness, and the microstructure. Both whey protein fractions contribute to the observed shift in gelation pH, although by a different mechanism. Obtaining gels with high gel hardness occurs most effectively when all denatured whey proteins are present as whey protein aggregates. It was observed that disulfide bridge exchange reactions during the acid-induced gelation at ambient temperature play an important role for both whey protein fractions. Additionally, disulfide interactions seem to occur between the aggregates and the casein micelles during the gel state. In this study, we show the development of a new approach for confocal scanning laser microscopy measurements—i.e., separate staining of the proteins in milk. By using this method, we were able to determine that, although whey protein aggregates are not linked to the casein micelles, they nevertheless gel at the same moment. This work adds to a better understanding of the role of denatured whey proteins during acid-induced gelation and could improve the effective use of whey proteins.     

Rheological characterization and electrokinetic phenomena of charged whey protein dispersions of defined sizes

A multi-step processing technique produced large colloidal particles from whey proteins, prompting instantaneous thickening upon hydration. Analysis of the rheological characteristics and zeta potential of the modified whey suspensions of defined particle sizes allowed investigation into the role of size on ingredient functionality. Preliminarily, the modified protein powders were sieved to achieve three size ranges, and analyzes were conducted on each of the three distributions and the non-sieved fractions. Following hydration, steady and oscillatory shear analyzes were performed using a controlled stress rheometer to determine rheological characteristics. Intrinsic viscosity was determined with a capillary viscometer and application of the Huggins equation. Zeta potential was calculated from colloidal electrophoretic mobility, measured with a ZetaPALS analyser. After thorough hydration, particle-size analysis revealed a size increase of >1.3 times for each fraction. When analysed on a protein basis, increasing particle size yielded an increase to intrinsic viscosity, flow behavior index, zero shear viscosity, and a decreased zeta potential and consistency coefficient. Knowledge of the interrelationship between zeta potential, rheological properties, and particle size of the modified whey ingredient will further advance an understanding of the functionality of this protein ingredient.     

酸度和钙质量分数对复原乳酶凝胶质地特性的影响

考察了不同酸度、钙质量分数调节后复原乳酶凝胶的质地特性,并对其质地变量进行了主成分分析。结果表明,不同工艺处理样品间的差异可以清晰地表现在两个新变量中。酸度、钙质量分数调整可对凝胶的表观黏度和持水力产生很大影响。此外,通过对照分析可以发现,不同前处理的复原乳样品具有与原料乳相近的凝胶特性。     

Gelation of casein micelles in ��-casein reduced milk prepared using membrane filtration

Pasteurized skim milk was subjected to membrane filtration using a molecular weight cut-off of 80 kDa and a plate and frame pilot scale system at temperatures below 10 °C. Via this process, transmission of whey proteins and ??-casein through the membrane was achieved. The milk was concentrated to two times (based on volume reduction), and whey protein-free permeate was added to return to the original volume fraction of casein micelles in milk. This diafiltration process was carried out four times, and the retentate obtained was nearly free of whey proteins and with approximately 20% of ??-casein removed. The same membrane filtration was also carried out at 25 °C to achieve transmission of whey protein but not of ??-casein, and to obtain whey protein-depleted milk without depletion of ??-casein.The gelling behaviour of these samples, reconstituted to the original casein volume fraction, was examined using rheology and diffusing wave spectroscopy. When compared to the original skim milk it was found that there were no statistically significant differences in gelation behaviour during acidification, but differences were noted in gelation time and final stiffness modulus for samples undergoing renneting. These differences were attributed mostly to the changes in ionic composition, as when the serum composition of the retentates was re-equilibrated against the original skim milk by dialysis; the gelation behaviour of the samples was comparable to that of skim milk. The results clearly indicate the importance of the milk's overall ionic balance in the early stages of aggregation of rennet-induced gelation of milk.     

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 whey and pea protein blends on the rheological and sensory properties of protein-based systems flavoured with cocoa

Whey and pea protein combined in different proportions (100W:0P, 75W:25P, 50W:50P, 25W:75P, 0W:100P) were used to prepare protein-based systems flavoured with cocoa and containing κ-carrageenan or κ-carrageenan/xanthan gum as thickeners. Steady and dynamic shear rheological properties of samples were measured at 10 °C and sensory differences were evaluated. Protein-based systems exhibited a shear-thinning flow behaviour that was fitted to the simplified Carreau model. Samples showed different viscoelastic properties, ranging from fluid-like to weak gel behaviour. For both types of system (with and without xanthan gum) viscosity, pseudoplasticity and elasticity rose on increasing the pea protein proportion in the blend. The sample with only whey protein obeyed the Cox-Merz rule, while in the rest of the samples complex viscosity was higher than apparent viscosity. Regarding sensory properties, the protein blend ratio mainly affected sample thickness, which rose as pea protein proportion increased. However, at the same time, the chocolate flavour and sweetness decreased and the off-flavour increased.     

Impact of protein self-assemblages on foam properties

The influence of dynamically heat-induced aggregates on whey protein foams was investigated as a function of the thermal treatment applied to WPI using a bubbling technique. The aim was to determine the interplay between the size/shape/proportion of the heat-induced aggregates and the properties of protein foams (formation and stability). Results showed that insoluble protein aggregates were highly branched and cohesive, whereas soluble aggregates were constituted by subunits, associated by hydrophobic bonds and formed by α-La and β-Lg monomers linked by disulfide bridges. Using the bubbling procedure, protein aggregates were shown to slow down significantly foam formation. However, the rate of foam formation remained nearly unchanged for wet foams when the amount of insoluble aggregates was inferior to 5% and when their size remained lower than 100 μm. Similarly, protein aggregates did not seem to affect the destabilisation kinetics of wet foams, regardless of amount, size, shape and proportion.     

Effect of storage time and temperature of milk protein concentrate (MPC85) on the renneting properties of skim milk fortified with MPC85

This study investigated the effect of storage temperature (20–50 °C) and time (0–60 days) on the renneting properties of milk protein concentrate with 85% protein (MPC85). Reconstituted skim milk was fortified with the MPC85 (2.5% w/w) and the renneting properties of the skim milk/MPC85 systems were investigated using rheology. It was found that the final complex modulus (final G∗) and the yield stress of the rennet-induced skim milk/MPC85 gels decreased exponentially with storage time of the MPC85 for storage temperatures greater than 20 °C, with a greater effect at the higher storage temperatures. Changes in the solubility of MPC85 with storage time were correlated with the rheological properties. The primary phase of renneting (cleavage of κ-casein) was not affected by the storage of the MPC85; hence the effect was related to the secondary stage of renneting (aggregation/coagulation of rennet-treated casein micelles). Using a temperature–time superposition method, a master curve was formed from the final G∗, yield stress and solubility results. This suggested that the same physical processes affected the solubility and rennet gelation properties of the milks. It is proposed that the MPC85 protein in rennet-treated skim milk/MPC85 solutions may transform from an interacting material, when solubility is high, to an inert or weakly interacting material, when solubility is low, and that this results in the reduced final G∗ and yield stress of the rennet gels when MPC85 is stored at elevated temperatures for long periods.     

Functional properties of whey protein concentrate texturized at acidic pH: Effect of extrusion temperature

Reactive supercritical fluid extrusion (RSCFX) process at acidic condition (pH 3.0) was used to generate texturized whey protein concentrate (TWPC) and the impacts of process temperature on product's physicochemical properties were evaluated. TWPC extruded at 50 and 70 °C formed soft-textured aggregates with high solubility than that extruded at 90 °C that formed protein aggregates with low solubility. Total free sulfhydryl contents and solubility studies in selected buffers indicated that TWPC is primarily stabilized by non-covalent interactions. Proteins texturized at 90 °C showed an increased affinity for 1-anilino-naphthalene-8-sulfonate (ANS) and a decreased affinity for cis-parinaric acid (CPA), indicating changes in protein structure. Water dispersion of TWPC at room temperature showed thickening function with pseudoplastic behavior. Secondary gelation occurred in TWPC obtained at 50 and 70 °C by heating the cold-set gels to 95 °C. TWPC texturized at 90 °C produced cold-set gels with good thermal stability. Compared to control, TWPC formed stable oil-in-water emulsions. Factors such as degree of protein denaturation and the balance of surface hydrophobicity and solubility influenced the heat- and cold-gelation and emulsifying properties of the protein ingredients. TWPC generated by low and high temperature extrusions can thus be utilized for different products requiring targeted physicochemical functionalities.     

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.     

Rheological,thermal and microstructural properties of whey protein isolate‐modified cassava starch mixed gels at different pH values

The objective of this study was to investigate the rheological, thermal and microstructural properties of whey protein isolate (WPI)‐hydroxypropylated cassava starch (HPCS) gels and WPI‐cross‐linked cassava starch (CLCS) gels at different pH values (5.75, 7.00 and 9.00). The rheological results showed that the WPI‐modified starch gels had greater storage modulus (G?) values than the WPI‐native cassava starch gels at pH 5.75 and 7.00. Differential scanning calorimetry curves suggested that the phase transition order of the WPI and modified starch changed as the pH increased. Scanning electron microscopy images showed that the addition of HPCS and CLCS contributed to the formation of a compact microstructure at pH 5.75 and 7.00. A comprehensive analysis showed that the gelling properties of the WPI‐modified starch were affected by the difference between the WPI denaturation temperature and modified starch gelatinisation temperature and by the granular properties of the modified starch during gelatinisation. These results may contribute to the application of WPI‐modified starch mixtures in food preparation.     

Interfacial dynamic properties of whey protein concentrate/polysaccharide mixtures at neutral pH

The effect of two non-surface active polysaccharides (sodium alginate, SA, and λ-carrageenan, λ-C) in the aqueous phase on the surface dynamic properties (dynamic surface pressure and surface dilatational properties) of a commercial milk whey protein concentrate (WPC) adsorbed film at the air–water interface has been studied. A whey protein isolate (WPI) was used as reference. The WPC and WPI concentration (at 1.0% wt), temperature (at 20 °C), pH (7), and ionic strength (at 0.05 M) were maintained constant, while the effect of polysaccharide (PS) was evaluated within the concentration range 0.0–1.0% wt. The surface dynamic properties of the adsorbed films were measured in an automatic pendant drop tensiometer. At short adsorption time and in the presence of PS, the rate of diffusion of WPC to the interface was affected by the interactions with PS in the aqueous phase, which could limit protein availability for the adsorption. On the other hand, at long-term adsorption, the magnitudes of the molecular penetration and configurational rearrangement rates of WPC in mixed systems (WPC/PS) reflected the viscoelastic characteristics of the adsorbed films. The attractive interactions between WPC and PS and/or the WPC aggregation in the presence of PS, which depend on the proper polysaccharide and its concentration in the aqueous phase, have an effect on the adsorption kinetic parameters, the amount of WPC adsorbed at the air–water interface, and the dilatational viscoelastic characteristics of WPC/PS mixed systems.     

Rheological, texture and sensory properties of low-fat mayonnaise with different fat mimetics

Application of whey protein isolate and low-methoxy pectin-based fat mimetics in mayonnaise was studied. Fat was partially substituted by different fat mimetics at levels of 50%, respectively, which the fat mimetics were referred to as PFM1 (microparticulate pectin gel), PFM2 (pectin weak-gel), and PFM3 (microparticulated combination of WPI and pectin). The full fat (Ff) (100% oil) mayonnaise without fat mimetic was used as a control experiment. Physicochemical, rheological, texture analysis, and sensory evaluation of the Ff and low fat (Lf) mayonnaises were performed. The results indicated that all Lf mayonnaises had significantly lower energy content, but higher water content than their Ff counterpart. In terms of texture, the formulation with pectin weak-gel as fat mimetic showed similar texture values as those of the Ff sample. Both Ff and Lf mayonnaises exhibited thixothopic shear thinning behaviour under steady shear tests and were rheologically classified as weak gels under small amplitude oscillatory shear tests. Sensory evaluation demonstrated that mayonnaises substituted with low-methoxy pectin were acceptable. This study shows good potential for pectin weak-gel and microparticulated pectin gel to be used as a fat mimetic in mayonnaise.     

Rheological investigation of soy protein hydrogels induced by Maillard-type reaction

Rheological measurements were used to determine the power law relationship between gel elastic modulus and protein concentration (solid volume fraction) for soy protein hydrogels. The slope calculated from the log–log plot was used to determine gel fractal dimension. The validity of the prefactor in the power law relationship was also studied. Soy protein at 6.5–9.5% was gelled using monofunctional (glyceraldehyde) or bifunctional (glutaraldehyde) Maillard cross-linkers with or without salt.Gel fractal dimension did not vary with cross-linker type or concentration, while prefactor values increased with cross-linker concentration and when salt was used. The invariability of the fractal dimension is believed related to the absence of effect of the cross-linking agents on protein ionic repulsions, while prefactor value is believed inversely proportional to the number of inter-floc particle–particle interactions in the three-dimensional network and to floc size. The prefactor value was thus shown to increase as cross-linking degree increased and gel lacunarity decreased.     

The effect of the pH at cooking on the properties of processed cheese spreads containing whey proteins

Processed cheese spreads were made with and without whey proteins under varying cooking pH conditions. The processed cheeses were cooked at one pH value and at the end of the cooking process the pH was adjusted to the final product pH of 5.7. The rheological properties and whey protein denaturation levels of the processed cheese spreads were measured. The rheological properties and texture of the processed cheeses containing whey proteins could be markedly modified by varying the cooking pH during processing, whereas those without whey proteins were unaffected. These textural modifications could not be explained solely by the changes in whey protein denaturation during cooking. It is proposed that the interactions of the whey proteins during cooking affect the processed cheese texture, and that these interactions are affected by the pH of the processed cheese during processing.     

Physicochemical and textural properties of heat-induced pea protein isolate gels

Chemical and thermal properties of pea protein isolates (laboratory prepared or native; PPIn and commercial; PPIc) and textural properties of heat-set gels obtained from pea protein isolates were compared with homologous soy protein isolates (laboratory prepared, or native; SPIn and commercial; SPIc). The protein banding pattern resulting from electrophoresis separation confirmed the presence of predominant storage proteins of pea and soy seeds in the respective protein products. PPIc and SPIc had lower nitrogen solubility than their native counterparts, likely due to their denaturated state which was further confirmed by the absence of distinct endotherms in these commercial materials compared to the laboratory prepared ones. Addition of NaCl at 1.0–2.0% (w/w) to PPIn and SPIn slurries increased thermal transition temperatures for both proteins.