Enhancement of the functional properties of whey protein by conjugation with maltodextrin under dry‐heating conditions

Conjugation of whey protein isolate (WPI) and maltodextrin (MD, dextrose equivalent of 6) was achieved by dry‐heating at an initial pH of 7.0, at 60 °C and 79% relative humidity, with WPI: MD6 ratio of 1:1, for up to 24 h. Conjugation was achieved with limited development of colour and advanced Maillard products on 24 h of heating. Conjugation increased the protein solubility at pH 4.5, by 7.1–8.5%, compared to the unheated and heated WPI controls. Conjugation of WPI with MD6 enhanced the stability and retention of clarity in protein solutions heated at 85 °C for 10 min with 50 mM added NaCl.     

Transglutaminase cross-linking to enhance elastic properties of soy protein hydrogels with intercalated montmorillonite nanoclay

To strengthen the network of bio-nanocomposite hydrogels, layered montmorillonite (MMT) nanoclay was intercalated by surface-coating with soy protein (SP) before mixing with 6% w/v SP for cross-linking by microbial transglutaminase (mTGase). Dynamic rheology was performed to study variables of NaCl and mTGase concentrations, with and without 1% w/v MMT. Without mTGase, the highest storage modulus (G′) was observed at 100 mM for samples without MMT, which was twice of the highest G′ for samples with MMT, at 200 mM NaCl. With mTGase, a shorter gelation time and a stronger hydrogel were observed at a higher enzyme level. Overall, the non-gelling 6% w/v SP dispersion was transformed to a hydrogel with G′ of 1099 Pa after addition of 100 mM NaCl and 1% SP-coated MMT and treatments by 6.25 U/g-protein mTGase for 2 h and heating/cooling steps. The integration of surface-coating and mTGase cross-linking is promising to improve properties of the nanocomposite system.     

Microemulsions as nanoreactors to produce whey protein nanoparticles with enhanced heat stability by thermal pretreatment

Native whey proteins (NWPs) may form gels or aggregates after thermal processing. The goal of this work was to improve heat stability of NWPs by incorporating protein solutions in nanoscalar micelles of water/oil microemulsions to form whey protein nanoparticles (WPNs) by thermal pretreatment at 90 °C for 20 min. The produced WPNs smaller than 100 nm corresponded to a transparent dispersion. The WPNs produced at NWP solution pH of 6.8 had a better heat stability than those produced at pH 3.5. The salt concentration (0–400 mM NaCl) in NWP solutions did not significantly change the size of corresponding WPNs. Compared to NWPs, the 5% (w/v) dispersion of WPNs at pH 6.8, 100 mM NaCl did not form a gel after heating at 80 °C for 20 min. The improved heat stability and reduced turbidity of WPNs may enable novel applications of whey proteins in beverages.     

Effect of dynamic heat treatment on the physical properties of whey protein foams

The influence of dynamically heat-induced aggregates on whey protein foams was investigated as a function of the thermal treatment applied with the aim of determining the optimal temperature for the production of heat-induced aggregates dedicated to foaming. The native protein solutions (2% w/v WPI; 50 mM NaCl) at neutral pH were heat-treated using a tubular heat exchanger between 70 °C and 100 °C. Protein denaturation and aggregation were followed by micro-differential scanning calorimetry, size exclusion chromatography, laser diffraction and dynamic light scattering. The protein solutions were whipped using a kitchen mixer to produce foams. Foam overrun, stability against drainage, texture and bubble size distribution were measured.     

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.     

Stabilization of soybean oil body emulsions using ι-carrageenan: Effects of salt,thermal treatment and freeze-thaw cycling

Soybean oil bodies and oleosins are known to be useful in foods and other emulsion systems. The influence of NaCl addition (0–500 mM), thermal processing (30, 60, 90 and 120 °C, 30 min), and freeze-thaw cycling (−20 °C, 24 h/30 °C, 2 h) on the stability of uncoated and ι-carrageenan coated soybean oil body emulsions at pH 3 and 7 was analyzed using particle electrical charge, particle size distribution, creaming stability and confocal laser scanning microscopy measurements. The stability of the uncoated emulsions to NaCl addition depended on pH, which was attributed to electrostatic screening effects. For NaCl, the uncoated emulsions were relatively stable from 0 to 150 mM at pH 3, but aggregated at ≥ 50 mM at pH 7; however, the ι-carrageenan coated emulsions at pH 3 and 7 were stable at all NaCl concentrations. The thermal stability of uncoated and ι-carrageenan coated emulsions may be relative to pH and holding temperature. Both uncoated and ι-carrageenan coated emulsions were stable at pH 7, whereas the uncoated emulsions at pH 3 became unstable when heated above 90 °C, but the ι-carrageenan coated emulsion droplets at pH 3 only extensively coalesced at 120 °C. The ι-carrageenan coated emulsions at pH 3 and 7 exhibited little droplet aggregation after three freeze-thaw cycles in the presence of sucrose. These results suggest that ι-carrageenan coated soybean oil body emulsions have similar or improved stability compared to uncoated emulsions and may be utilized as functional soy products in the food and other industries.     

Comparison of properties of oil-in-water emulsions stabilized by coconut cream proteins with those stabilized by whey protein isolate

Coconut cream protein (CCP) fractions were isolated from coconuts using two different isolation procedures: isoelectric precipitation (CCP1-fraction) and freeze–thaw treatment (CCP2-fraction). The ability of these protein fractions to form and stabilize oil-in-water emulsions was compared with that of whey protein isolate (WPI). Protein solubility was a minimum at ∼pH 4, 4.5 and 5 for CCP1, CCP2, and WPI, respectively, and decreased with increasing salt concentration (0–200 mM NaCl) for the coconut proteins. All of the proteins studied were surface active, but WPI was more surface active than the two coconut cream proteins. The two coconut cream proteins were used to prepare 10 wt% corn oil-in-water emulsions (pH 6.2, 5 mM phosphate buffer). CCP2 emulsions had smaller mean droplet diameters (d₃₂ ≈ 2 μm) than CCP1 emulsions (d₃₂ ≈ 5 μm). Corn oil-in-water emulsions (10 wt%) stabilized by 0.2 wt% CCP2 and WPI were prepared with different pH values (3–8), salt concentrations (0–500 mM NaCl) and thermal treatments (50–90 °C for 30 min). Considerable droplet flocculation occurred in the emulsions near the isoelectric point of the proteins: CCP2 (pH ∼ 4.3); WPI (pH ∼ 4.8). Emulsions with monomodal particle size distributions, small mean droplet diameters, and good creaming stability could be produced at pH 7 for WPI, but CCP2 produced bimodal distributions at this pH. The CCP2 and WPI emulsions remained relatively stable to droplet aggregation and creaming at NaCl concentrations ⩽50 and ⩽100 mM, respectively. In the absence of salt, both CCP2 and WPI emulsions were quite stable to thermal treatments (50–90 °C for 30 min).     

Combined effect of dynamic heat treatment and ionic strength on the properties of whey protein foams – Part II

The aim was to investigate the effect of dynamic thermal treatment in a tubular heat exchanger on the denaturation and foaming properties of whey proteins, such as overrun, foam stability and texture. A 2% w/v WPI solution (pH 7.0), with and without NaCl addition (100 mM), was submitted to heat treatment at 100 °C. The results demonstrated that heat treatment slightly reduced overrun, whereas NaCl and heat treatment improved foam stability, enhanced texture and provided smaller bubble diameters with more homogeneous bubble size distributions in foams. The foaming properties of proteins, especially stability, were shown to depend not only on the amount of protein aggregates, but also on their size. While insoluble aggregates (larger than 1 μm diameter) accelerated drainage, soluble aggregates (about 200 nm diameter) played a key role on the stabilization of gas–liquid interfaces.     

Improvement of functional properties of whey protein isolate through glycation and phosphorylation by dry heating

Whey protein isolate (WPI) was glycated with maltopentaose (MP) through the Maillard reaction, and the MP-conjugated WPI (MP-WPI) was then phosphorylated by dry heating in the presence of pyrophosphate. Glycation occurred efficiently, and the sugar content of WPI increased approximately 19.9% through the Maillard reaction. The phosphorylation of MP-WPI was enhanced with an increase in the dry-heating time from 1 to 5 d, and the phosphorus content of WPI increased approximately 1.05% by dry heating at pH 4.0 and 85°C for 5 d in the presence of pyrophosphate. The electrophoretic mobility of WPI increased with an increase in the phosphorylation level. The stability of WPI against heat-induced insolubility at pH 7.0 was improved by conjugation with MP alone, and further improved by phosphorylation. Although the emulsifying activity of WPI was barely affected by glycation and phosphorylation, the emulsifying stability of phosphorylated MP-WPI (5 d), was 2.2 times higher than that of MP-WPI. Gelling properties such as hardness, resiliency, and water-holding capacity of heat-induced WPI gel were markedly improved, and the gel was rendered transparent by phosphorylation. The calcium phosphate-solubilizing ability of WPI was enhanced by phosphorylation. These results suggested that phosphorylation by dry heating in the presence of pyrophosphate after conjugation with MP is a useful method for improving the functional properties of WPI.     

Effect of thermal treatments on the properties of coconut milk emulsions prepared with surface-active stabilizers

Previously we have demonstrated improved stability of coconut milk emulsions homogenized with various surface-active stabilizers, i.e., 1 wt% sodium caseinate, whey protein isolate (WPI), sodium dodecyl sulfate (SDS), or polyoxyethylene sorbitan monolaurate (Tween 20) [Tangsuphoom, N., & Coupland, J. N. (2008). Effect of surface-active stabilizers on the microstructure and stability of coconut milk emulsions. Food Hydrocolloids, 22(7), 1233–1242]. This study examines the changes in bulk and microstructural properties of those emulsions following thermal treatments normally used to preserve coconut milk products (i.e., −20 °C, −10 °C, 5 °C, 70 °C, 90 °C, and 120 °C). Calorimetric methods were used to determine the destabilization of emulsions and the denaturation of coconut and surface-active proteins. Homogenized coconut milk prepared without additives was destabilized by freeze–thaw, (−20 °C and −10 °C) but not by chilling (5 °C). Samples homogenized with proteins were not affected by low temperature treatments while those prepared with surfactants were stable to chilling but partially or fully coalesced following freeze–thaw. Homogenized coconut milk prepared without additives coalesced and flocculated after being heated at 90 °C or 120 °C for 1 h in due to the denaturation and subsequent aggregation of coconut proteins. Samples emulsified with caseinate were not affected by heat treatments while those prepared with WPI showed extensive coalescence and phase separation after being treated at 90 °C or 120 °C. Samples prepared with SDS were stable to heating but those prepared with Tween 20 completely destabilized by heating at 120 °C.     

Improvement of the functional properties of whey protein hydrolysate by conjugation with maltodextrin

The impact of conjugation with maltodextrin on selected functional properties (i.e., solubility and thermal stability) of intact whey protein isolate (WPI) and whey protein hydrolysate (WPH) was determined. Conjugation of WPI and WPH (degree of hydrolysis 9.3%) with maltodextrin (MD) was achieved by heating solutions of 5% WPI or WPH with 5% MD, initial pH 8.2, at 90 °C for up to 24 h. The WPH had 55.4% higher levels of available amino groups compared with the WPI, which contributed to more rapid and extensive conjugation of WPH-MD, compared with WPI-MD. The WPI-MD and WPH-MD solutions heated for 8 h had significantly higher (P < 0.05) protein solubility than the respective WPI and WPH heated control solutions, in the pH range 4.0–5.0. Conjugation of WPI and WPH with MD enhanced the stability to heat-induced changes, such as turbidity development, gelation or precipitation, in the presence of 40 mm added NaCl.     

Influence of calcium-binding salts on heat stability and fouling of whey protein isolate dispersions

The effect of the calcium-binding salts (CBS), trisodium citrate (TSC), tripotassium citrate (TPC) and disodium hydrogen phosphate (DSHP) at concentrations of 1–45 mm on the heat stability and fouling of whey protein isolate (WPI) dispersions (3%, w/v, protein) was investigated. The WPI dispersions were assessed for heat stability in an oil bath at 95 °C for 30 min, viscosity changes during simulated high-temperature short-time (HTST) and fouling behaviour using a lab-scale fouling rig. Adding CBS at levels of 5–30 mm for TSC and TPC and 25–35 mm for DSHP improved thermal stability of WPI dispersions by decreasing the ionic calcium (Ca²⁺) concentration; however, lower or higher concentrations destabilised the systems on heating. Adding CBS improved heat transfer during thermal processing, and resulted in lower viscosity and fouling. This study demonstrates that adding CBS is an effective means of increasing WPI protein stability during HTST thermal processing.     

Effect of microbial transglutaminase treatment on thermal stability and pH-solubility of heat-shocked whey protein isolate

Whey protein isolate (WPI) dispersions (5% protein, pH 7.0) were subjected to heat-shock at 70 °C for 1, 5 and 10 min. The heat-shocked WPI dispersions were treated with microbial transglutaminase (MTGase) enzyme, and thermal properties and pH-solubility of the treated proteins were investigated. Heat-shocking of WPI for 10 min at 70 °C increased the thermal denaturation temperature (T_d) of β-lactoglobulin in WPI by about 1.5 °C. MTGase treatment (30 h, 37 °C) of the heat-shocked WPI significantly increased the T_d of β-lactoglobulin by about 6.3–7.3 °C when compared with heat-shocked only WPI at pH 7.0. The T_d increased by about 13–15 °C following pH adjustment to 2.5; however, the T_d of heat-shocked WPI was not substantially different from heat-shocked and MTGase-treated WPI at pH 2.5. Both the heat-shocked and the heat-shocked-MTGase-treated WPI exhibited U-shaped pH-solubility profiles with minimum solubility at pH 4.0–5.0. However, the extent of precipitation of MTGase-treated WPI samples at pH 4.0–5.0 was much greater than all heat-shocked and native WPI samples. The study revealed that while MTGase cross-linking significantly enhanced the thermal stability of β-lactoglobulin in heat-shocked WPI, it caused pronounced precipitation at pH 4.0–5.0 via decreasing the hydrophilic/hydrophobic ratio of the water-accessible protein surface.     

Whey protein nanoparticles prepared with desolvation with ethanol: Characterization,thermal stability and interfacial behavior

Whey protein isolate (WPI) nanoparticles were prepared by diluting an alkaline solution of protein in ethanol at concentrations varying between 50 and 80%. The nanoparticles were then immediately diluted in buffer. While the nanoparticles were not stable at pH 7, they showed no changes in size when diluted at pH 3. When 75–80% ethanol was added during preparation, the size of the WPI nanoparticles ranged between 10 and 100 nm, with no change in size after dilution and storage at pH 3 for 96 h at 22 °C. When heating was applied, particle aggregation occurred, and large aggregates (>1 μm) were observed at temperatures > 60 °C. The particle size of the heat-induced aggregates could be reduced by homogenization. The nanoparticles prepared by desolvation showed interfacial pressure values similar to those of the corresponding protein solutions, indicating similar interfacial properties and the potential to be used to stabilize emulsions but as supramolecular aggregates of WPI.     

Rheology and Oxidative Stability of Whey Protein Isolate-Stabilized Menhaden Oil-in-Water Emulsions as a Function of Heat Treatment

ABSTRACT:  Menhaden oil-in-water emulsions (20%, v/v) were stabilized by 2 wt% whey protein isolate (WPI) with 0.2 wt% xanthan gum (XG) in the presence of 10 mM CaCl₂ and 200 μM EDTA at pH 7. Droplet size, lipid oxidation, and rheological properties of the emulsions were investigated as a function of heating temperature and time. During heating, droplet size reached a maximum at 70 °C and then decreased at 90 °C, which can be attributed to both heating effect on increased hydrophobic attractions and the influence of CaCl₂ on decreased electrostatic repulsions. Combination of effects of EDTA and heat treatment contributed to oxidative stability of the heated emulsions. The rheological data indicate that the WPI/XG-stabilized emulsions undergo a state transition from being viscous like to an elastic like upon substantial thermal treatment. Heating below 70 °C or for less than 10 min at 70 °C favors droplet aggregation while heating at 90 °C or for 15 min or longer at 70 °C facilitates WPI adsorption and rearrangement. WPI adsorption leads to the formation of protein network around the droplet surface, which promotes oxidative stability of menhaden oil. Heating also aggravates thermodynamic incompatibility between XG and WPI, which contributes to droplet aggregation and the accumulation of more WPI around the droplet surfaces as well.     

Controlling denaturation and aggregation of whey proteins during thermal processing by modifying temperature and calcium concentration

Whey protein isolate solutions (8.00 g protein/100 g; pH 6.8) were treated for 2 min at 72, 85 or 85 °C with 2.2 mM added calcium Ca to produce four whey protein systems: unheated control (WPI‐UH), heated at 72 °C (WPI‐H72), heated at 85 °C (WPI‐H85) or heated at 85 °C with added Ca (WPI‐H85Ca). Total levels of whey protein denaturation increased with increasing temperature, while the extent of aggregation increased with the addition of Ca, contributing to differences in viscosity. Significant changes in Ca ion concentration, turbidity and colour on heating of WPI‐H85Ca, compared to WPI‐UH, demonstrated the role of Ca in whey protein aggregation.     

Viscoelastic behavior and microstructure of aqueous mixtures of cross-linked waxy maize starch,whey protein isolate and κ-carrageenan

The viscoelasticity and microstructure of mixtures of cross-linked waxy maize starch (CH10), whey protein isolate (WPI) and κ-carrageenan (κC) at pH 7.0 with 100 mM NaCl were investigated by oscillatory rheometry and confocal laser scanning microscopy (CLSM). Mixtures were heated to 90 °C (1.5 °C/min), held for 10 min at this temperature and cooled. Within the range of concentrations studied, CH10 swollen granules reinforced WPI and κC networks. The mechanical behavior of the three-component mixtures was modified by different WPI concentrations, but κC governed the overall response due to its gelling ability. CLSM images of three-component mixtures showed particulate systems in which swollen starch granules are surrounded by κC and WPI. CH10 granules were immersed in a single phase and a separate phase of κC and WPI, for low and high concentrations of these components, respectively. Therefore, it is possible to obtain two- and three-phase mixtures.     

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.     

Food-grade microemulsions,nanoemulsions and emulsions: Fabrication from sucrose monopalmitate & lemon oil

Sucrose monopalmitate (SMP) is a non-toxic, biodegradable, non-ionic surfactant suitable for use in foods and beverages. This study aimed to establish conditions where stable microemulsions, nanoemulsions or emulsions could be fabricated using SMP as a surfactant and lemon oil as an oil phase. Emulsions (r > 100 nm) or nanoemulsions (r < 100 nm) were formed at low surfactant-to-oil ratios (SOR < 1) depending on homogenization conditions, whereas microemulsions (r < 10 nm) were formed at higher ratios (SOR > 1). The impact of simple mixing, thermal treatment, and homogenization on the formation of the different colloidal systems was investigated. Blending/heating was needed to produce microemulsions or emulsions, whereas blending/heating/homogenization was needed to produce nanoemulsions. The impact of environmental stresses (pH, ionic strength, temperature) on the functional performance of nanoemulsions and microemulsions was examined. Relatively stable nanoemulsions could be formed at pH 6 and 7 and stable microemulsions at pH 5 and 6, but extensive particle growth/aggregation occurred at lower and higher pH values. Microemulsions were relatively stable to salt addition (0–200 mM NaCl), but nanoemulsions exhibited droplet aggregation/growth at ≥50 mM NaCl after 1 month storage at pH 7. Microemulsions formed gels at low temperatures (5 °C), were stable at ambient temperatures (23 °C), and exhibited particle growth at elevated temperatures (40 °C). Nanoemulsions were stable at refrigerator (5 °C) and ambient (23 °C) temperatures, but exhibited coalescence at elevated temperatures (40 °C). This study provides important information for optimizing the application of sucrose monoesters to form colloidal dispersions in food and beverage products.     

Influence of EDTA and citrate on thermal stability of whey protein stabilized oil-in-water emulsions containing calcium chloride

The influence of calcium ions and chelating agents on the thermal stability of model nutritional beverages was examined. Oil-in-water emulsions (6.94% (w/v) soybean oil, 0.35% (w/v) WPI, 0.02% (w/v) sodium azide, 20 mM Tris buffer, 0–10 mM CaCl₂, and 0–40 mM EDTA or citrate, pH 7.0) were stored at temperatures between 30 and 120 °C for 15 min. The particle size, particle charge, creaming stability, rheology, and free-calcium concentration of the emulsions were then measured. In the absence of chelating agents, appreciable droplet aggregation occurred in emulsions held at temperatures from 80 to 120 °C, which led to increased emulsion particle diameter, shear-thinning behavior, apparent viscosity, and creaming instability. Addition of chelating agents to the emulsions prior to heating decreased, but did not prevent, droplet aggregation in the emulsions. EDTA was more effective than citrate in decreasing droplet aggregation. Heat treatment increased the amount of chelating agents required to prevent droplet aggregation in the emulsions. Free-calcium concentration and droplet surface potential was independent of heat-treatment temperature, indicating that the performance of the chelating agents in binding calcium ions was not affected by the heat treatment. It was suggested that increased hydrophobic attractive interactions between the droplets occurred during heating, which induced droplet aggregation.