Emulsification of Commercial Dairy Proteins with Exhaustively Washed Muscle

The addition of dairy proteins to exhaustively washed chicken breast muscle improved the emulsion stability in heated cream layers (emulsions) containing whey protein concentrate (WPC) or whey protein isolate (WPI). The initial weight of the heated cream layers made with WPC or WPI was heavier than those for sodium caseinate (CNate) or milk protein isolate (MPI). The addition of CNate or MPI resulted in decreased emulsion stability and increased inhibition of myosin heavy chain and actin participation in the emulsion formation compared to WPC or WPI.     

Influence of xanthan gum on the formation and stability of sodium caseinate oil-in-water emulsions

Studies have been made of the changes in droplet sizes, surface coverage and creaming stability of emulsions formed with 30% (w/w) soya oil, and aqueous solution containing 1 or 3% (w/w) sodium caseinate and varying concentrations of xanthan gum. Addition of xanthan prior to homogenization had no significant effect on average emulsion droplet size and surface protein concentration in all emulsions studied. However, addition of low levels of xanthan (≤0.2 wt%) caused flocculation of droplets that resulted in a large decrease in creaming stability and visual phase separation. At higher xanthan concentrations, the creaming stability improved, apparently due to the formation of network of flocculated droplets. It was found that emulsions formed with 3% sodium caseinate in the absence of xanthan showed extensive flocculation that resulted in very low creaming stability. The presence of xanthan in these emulsions increased the creaming stability, although the emulsion droplets were still flocculated. It appears that creaming stability of emulsions made with mixtures of sodium caseinate and xanthan was more closely related to the structure and rheology of the emulsion itself rather than to the rheology of the aqueous phase.     

Emulsifying Capacity and Emulsion Stability of Milk Proteins and Corn Germ Protein Flour

Comparative studies of emulsifying capacity (EC) and emulsion stability (ES) of corn germ protein flour (CGPF), nonfat dry milk (NFDM), whey protein concentrates (WPC), and sodium caseinate (SC) were carried out using response surface methodology. CGPF was an effective stabilizer of fat-protein-water emulsions. Tested proteins showed the following trend in EC and ES: SC > WPC = NFDM > CGPF. EC and ES increased with increased concentration of protein. No effect of incubation temperature on EC and ES of these proteins was detected. All protein samples showed a maximum EC at high pH (near 8.0) and 1.0% concentration under test conditions, except NFDM (about pH 7.0). Minimal pH effect on ES was found.     

Stability of model emulsions prepared using whey and muscle proteins

The amount of water (SW) and oil (SO) separated from model emulsions and emulsion stability (ES) of these emulsions prepared from corn oil and of fluid whey, total muscle protein (TMP), whey + TMP and sarcoplasmic protein (SP) were examined. The SW value of whey + TMP emulsions was lower (26,33%) than that of TMP (31,33%), SP (39,0%) practically the same as that of whey only. However, the SO value of whey emulsions was higher (9,40%) than that of muscle protein emulsions (0,0%). It was found that there was no oil separation in whey + TMP emulsions. Whey proteins had the lowest ES (64,6 ± 0,96) among the proteins studied. Nevertheless, whey + TMP emulsions had the highest ES (73,67 ± 0,58).     

Effect of high-methoxy pectin on properties of casein-stabilized emulsions

We report on the effect of high-methoxy pectin on the stability and rheological properties of fine sunflower oil-in-water emulsions prepared with α_s1-casein, β-casein or sodium caseinate. The aqueous phase was buffered at pH 7.0 or 5.5 and the ionic strength was adjusted with sodium chloride in the range 0.01–0.2 M. Average emulsion droplet sizes were found to be slightly larger at the lower pH and/or with pectin present during emulsification. Analysis of the serum phase after centrifugation indicated that some pectin becomes incorporated into the interfacial layer at pH 5.5 but not at pH 7.0. This was also supported by electrophoretic mobility measurements on protein-coated emulsion droplets and surface shear viscometry of adsorbed layers at the planar oil–water interface. A low pectin concentration (0.1 wt%) was found to give rapid serum separation of moderately dilute emulsions (11 vol% oil, 0.6 wt% protein) and highly pseudoplastic rheological behaviour of concentrated emulsions (40 vol% oil, 2 wt% protein). We attribute this to reversible depletion flocculation of protein-coated droplets by non-adsorbed pectin. At ionic strength below 0.1 M, the initial average droplet sizes, the creaming behaviour, and the rheology were found to be similar for emulsions made with either of the individual caseins (α_s1 and β) or with sodium caseinate. At higher ionic strength, however, whereas emulsions containing β-casein or sodium caseinate were stable, the corresponding α_s1-casein emulsions exhibited irreversible salt-induced flocculation which was not inhibited by the presence of the pectin.     

Rheological properties of emulsions containing milk proteins mixed with xanthan gum

Oil in water emulsions (30% w/w) containing mixtures of milk proteins with xanthan gum were rheologically characterized at ambient temperature and the evolution of their properties was measured during a month under cold storage. The milk proteins used were sodium caseinate and whey concentrate at 2% mixed with xanthan gum at 0.3% or 0.5%. Emulsions properties were compared to those of respective aqueous systems and in general showed same rheological behaviour as their respective aqueous system, however, emulsions presented higher consistency index, due to oil droplets concentration. The flow behaviour index showed a small variation, increasing its value slightly. The consistency of emulsions with xanthan was similar, independently of the milk protein used, confirming that xanthan rheology predominates on emulsion rheology.     

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

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

Whey protein–carboxymethylcellulose interaction in solution and in oil-in-water emulsion systems. Effect on emulsion stability

Carboxymethylcellulose (CMC) was used as coagulation aid to precipitate the whey proteins from defatted milk serum and the ability of the resulting whey protein concentrate (WPC, protein content: 63.69%) to aid in the physicochemical stabilization of oil-in-water emulsions, during ageing or following the application of heat or freeze–thaw treatment, was investigated, along with the stability of emulsion systems prepared with a commercial whey protein isolate. The stability of WPC emulsions against droplet flocculation and creaming, and to a lesser extent against droplet coalescence, depended on the presence of the CMC molecules in the emulsion continuous phase and the extent of adsorbed protein–polysaccharide interactions as affected by the emulsion pH. Studies on whey protein–CMC interaction were conducted, both in biopolymer mixture solutions and emulsion systems, by applying zeta potential measurement and viscometry techniques. These results were combined with data on protein surface hydrophobicity and on methylene blue-binding ability of CMC molecules and indicated that whey protein–CMC interaction may take place in solution, both at neutral as well as at acidic environments, leading, depending on pH, to the formation of soluble or non-soluble amphiphilic conjugates. In emulsion systems, however, conjugate formation is observed only at relatively acidic pH environments, probably because at a neutral or at a slightly acidic pH whey protein adsorption to the emulsion droplet surface and molecular unfolding does not favour protein–polysaccharide interaction.     

Creaming Stability of Oil-in-Water Emulsions Formed with Sodium and Calcium Caseinates

Creaming stability of emulsions formed with calcium caseinate, determined after storage of emulsions at 20 °C for 24 h, increased gradually with an increase in protein concentration from 0.5% to 2.0%; further increases in caseinate concentration had much less effect. In contrast, the creaming stability of sodium caseinate emulsions showed a decreased with an increase in protein concentration from 0.5% to 3.0%. Confocal laser micrographs of emulsions formed with >2% sodium caseinate showed extensive flocculation of oil droplets with the appearance of a network structure. However, emulsions formed with calcium caseinate or emulsions formed with low concentrations of sodium caseinate (0.5% and 1.0%) were homogenous with no sign of flocculation.     

Effect of protein oxidation on kinetics of droplets stability probed by microrheology in O/W and W/O emulsions of whey protein concentrate

Whey protein concentrate (WPC) was oxidized by peroxyl radicals derived from 2,2′-azobis (2-amidinopropane) dihydrochloride (AAPH) and the kinetics of droplet stability in O/W and W/O emulsions stabilized by oxidized WPC were evaluated by studying the micro-rheology. Degrees of protein oxidation were indicated by carbonyl concentration and emulsion types were distinguished by fluorescence microscopy. Oxidation resulted in free sulfhydryl groups degradation and surface hydrophobicity decrease. Moderate protein oxidation promoted to form diminutive droplets, which aggregated quickly to gel-network structure and decreased the motion rate of droplets, leading to the increased elasticity and viscosity, which led to better stability. Over-oxidation underwent severe droplet aggregation and sediment with increased motion rate, which resulted in instability of emulsions. The W/O emulsions of oxidized WPC were more inclined to block the motion of droplets and form a stable structure with higher viscosity, compared with the O/W emulsions.     

Oil-in-Water Emulsions Stabilized by Sodium Caseinate or Whey Protein Isolate as influenced by Glycerol Monostearate

Competitive adsorption between glycerol monostearate (GMS) and whey protein isolate (WPI) or sodium caseinate was studied in oil-in-water emulsions (20 wt % soya oil, deionized water, pH 7). Addition of GMS resulted in partial displacement of WPI or sodium caseinate from the emulsion interface. SDS-PAGE showed that GMS altered the adsorbed layer composition in sodium caseinate stabilized emulsions containing < 1.0 wt % protein. Predominance of β-casein at the interface in the absence of surfactant was reduced in the presence of GMS. The distribution of α-lactalbumin and β-lactoglobulin between the aqueous bulk phase and the fat surface in emulsions stabilized with WPI was independent of the concentration of added protein or surfactant.     

Functional characterization of protein stabilized emulsions: Emulsifying behaviour of proteins in a valve homogenizer

Protein stabilised emulsions have been prepared in a valve homogeniser incorporated into a recirculating emulsification system, where the power input and number of passes have been varied. The food proteins studied were a soy-bean protein isolate, a whey protein concentrate (WPC) and a sodium caseinate. The emulsions obtained were characterized in terms of particle size distribution and amount of protein adsorbed on to the fat surface (protein load). Generally, the final fat surface area of the emulsions obtained increases more as a function of power input than as a function of number of passes. Distribution width, c_s, decreases mostly with increasing power supply and number of passes, but at the highest power input c_s increases. The protein load on the fat globules is largely determined by the fat surface area and by the type of protein adsorbed. The soy proteins give a high protein load and the caseinates give a low protein adsorption at small fat surface areas created. This relation is reversed at large surface areas of the fat globules. The relation between percentage protein adsorbed from bulk as a function of surface area suggests that the caseinates mainly cover the newly created interface by adsorption from the bulk, whereas the soy proteins fulfil this task mostly by spreading at the interface. Salt addition to 0.2M-NaCl enhances protein adsorption at the fat globule interface in the case of soy protein and caseinate, but for the whey proteins protein load is higher in distilled water.     

Rheological and equilibrium properties of milk proteins and tara gum mixtures

The stability of aqueous purified tara gum (TG) mixtures with sodium caseinate (NaCas) and whey protein concentrate (WPC) was investigated. Phase diagram of TG–milk proteins mixtures was obtained by measuring the sedimented fraction or the rheological parameters. The rheological behaviour was also evaluated as function of NaCas and WPC concentration, considering the zero-shear rate viscosity and the relaxation time as response variables. The rheological parameters were determined by rotational rheometry. The studied solutions presented a meta-stable condition provided by the increase in viscosity, while mixtures with NaCas at low TG concentrations sedimentation by volume exclusion occurred. The rheological behaviour of NaCas and WPC aqueous solutions approached the Newtonian and shear-thickening model, respectively. The mixtures with TG showed a pseudoplastic behaviour, approaching the Cross model. The results indicated interaction between TG and the studied proteins, expressed by an increase of zero-shear viscosity and the relaxation time at higher gum concentration.     

Survival of Bifidobacterium longum 1941 microencapsulated with proteins and sugars after freezing and freeze drying

Five types of proteins and three types of sugars were examined for their effectiveness in protecting B. longum after freeze drying, including their acid and bile tolerance, surface hydrophobicity, retention of β-glucosidase, lactate dehydrogenase and adenosine triphosphatase. Sodium caseinate 12%, whey protein concentrate 12%, sodium caseinate:whey protein concentrate 6%:6%, skim milk 12%, or soy protein isolate 12% was combined with glycerol (3% w/v), mannitol (3% w/v) or maltodextrin (3% w/v). Fifteen emulsion systems containing sugars were obtained. Concentrated B. longum 1941 was incorporated into each emulsion system at a ratio of 1:4 (bacteria:emulsion). All the mixtures were then freeze dried. Water activity (a_w) of freeze dried microcapsules was in the range of 0.30 to 0.35. WPC–CAS GLY provided high stability of bacteria (99.2%) during freezing, while high stability of cells after freeze drying and during exposure to acid and bile environment was achieved when CAS–MAN was applied (97.4%, 81.6% and 99.3%, respectively). High retention of β-glu of freeze-dried bacteria was achieved using SM–MAN as protectant (94.6%). ATPase and LDH were successfully retained by SM–GLY (94.9 and 83.6%, respectively) but there was no significant difference in protection effect using CAS–MAN (93.8 and 82.6%, respectively). Overall, milk proteins were superior to SPI and sugar alcohols provided more protection than MD.     

Interactions between Whey Protein Isolate and Soy Protein Fractions at Oil–Water Interfaces: Effects of Heat and Concentration of Protein in the Aqueous Phase

ABSTRACT:  The 2 main storage proteins of soy—glycinin (11S) and β-conglycinin (7S)—exhibit unique behaviors during processing, such as gelling, emulsifying, or foaming. The objective of this work was to observe the interactions between soy protein isolates enriched in 7S or 11S and whey protein isolate (WPI) in oil–water emulsion systems. Soy oil emulsion droplets were stabilized by either soy proteins (7S or 11S rich fractions) or whey proteins, and then whey proteins or soy proteins were added to the aqueous phase. Although the emulsifying behavior of these proteins has been studied separately, the effect of the presence of mixed protein systems at interfaces on the bulk properties of the emulsions has yet to be characterized. The particle size distribution and viscosity of the emulsions were measured before and after heating at 80 and 90 °C for 10 min. In addition, SDS-PAGE electrophoresis was carried out to determine if protein adsorption or exchanges at the interface occurred after heating. When WPI was added to soy protein emulsions, gelling occurred with heat treatment at WPI concentrations >2.5%. In addition, whey proteins were found adsorbed at the oil–water interface together with 7S or 11S proteins. When 7S or 11S fractions were added to WPI-stabilized emulsions, no gelation occurred at concentrations up to 2.5% soy protein. In this case also, 7S or 11S formed complexes at the interface with whey proteins during heating.     

High Pressure Effects on Emulsifying Behavior of Whey Protein Concentrate

Oil-in-water emulsions (0.4 wt% protein, 20 vol%n-tetradecane, pH 7) prepared with solutions of pressure-treated (up to 800 MPa) whey protein concentrate (WPC) as emulsifier give a broader droplet-size distribution than emulsions made with native untreated protein. There was a decrease in emulsifying efficiency with increasing applied pressure and treatment time. In contrast, pressure treatment of corresponding WPC emulsions made with the native protein had little effect on emulsion stability. In the pressure-treated emulsion the protein is probably already conformationally modified so that pressure has little additional effect. However, in solution the native structure of the whey protein is modified by pressure resulting in loss of emulsifying efficiency.     

Effects of varying time/temperature-conditions of pre-heating and enzymatic cross-linking on techno-functional properties of reconstituted dairy ingredients

The effects of varying time/temperature-conditions of pre-heating and cross-linking with transglutaminase (TG) on the functional properties of reconstituted products from skim milk, WPC and sodium caseinate was analyzed. The degree of cross-linking (DC) of skim milk proteins could be increased from 54.4% to 70.5% by varying process conditions. Thereby the water-holding capacity (WHC) increased from 10% to 20%, while the heat stability decreased. The burning-on was lower than that of the non-treated products at optimum pre-heating conditions (90 °C/30 s). Using sodium caseinate as substrate for TG the DC increased from 39.2% to 100% due to the improvement of the process. As a result the WHC increased by 30% and the heat stability up to 380%. However, the burning-on of casein increased as well. TG-treated sodium caseinate started to gel at 10% protein, whereas untreated sodium caseinate gelled not before 15% protein. The WHC of enzyme-treated whey proteins was lowered. The heat stability of WPC could be doubled by TG-treatment, and the burning-on of the products was, especially at optimum pre-heating conditions, less pronounced. The degree of denaturation of TG-treated whey proteins was 2–5% higher than that of untreated samples.     

Transglutaminase cross-linking kinetics of sodium caseinate is changed after emulsification

Enzymatic cross-linking is an important method of modifying the structure of food products to control their texture and stability. In this paper we look at the effect that adsorption to the oil–water interface of triglyceride oil-in-water emulsion has on rates of cross-linking of sodium caseinate by microbial transglutaminase. The kinetics of cross-linking has also been assessed for the individual casein proteins within the caseinate. In solution the rates were α_s2-casein > β-casein > α_s1-casein > κ-casein. This order is not as expected given the rheomorphic nature of the proteins and the number of glutamine and lysine residues in each protein. In particular, the α_s1-casein was cross-linked much more slowly than expected. When sodium caseinate was adsorbed to an emulsion the rates for all constituent caseins were decreased but the cross-linking rate for α_s1-casein was markedly reduced, indicating the most significant change in accessibility following adsorption. This knowledge will facilitate optimal production of cross-linked emulsions for use in future studies aimed at engineering emulsions with improved nutritional quality.     

Impact of whey protein isolates and concentrates on the formation of protein nanoparticles‐stabilised Pickering emulsions

Whey protein nanoparticles (NPs) were prepared by heat‐induced method. The influences of whey protein isolates (WPIs) and concentrates (WPCs) on the formation of NPs were first investigated. Then Pickering emulsions were produced by protein NPs and their properties were evaluated. After heat treatment, WPC NPs showed larger particle size, higher stability against NaCl, lower negative charge and contact angle between air and water. Dispersions of WPC NPs appeared as higher turbidity and viscosity than those of WPI NPs. The interfacial tension of WPC NPs (~7.9 mN/m at 3 wt% NPs) was greatly lower than that of WPI NPs (~12.1 mN/m at 3 wt% NPs). WPC NPs‐stabilised emulsions had smaller particle size and were more homogeneous than WPI NPs‐stabilised emulsions. WPC NPs‐stabilised emulsions had higher stability against NaCl, pH and coalescence during storage.     

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.