Heat-induced destabilization of oil-in-water emulsions formed from hydrolyzed whey protein.

The emulsifying ability, heat stability, and coalescence stability of oil-in-water emulsions prepared with whey protein of varied degrees of hydrolysis (DH), and at varied protein contents, was studied. Whey protein hydrolysates (WPH) with a DH of 4% and 10% had poorer emulsifying ability than non-hydrolyzed whey protein concentrate (WPC), but were more heat stable. Increasing DH between 10 and 27% improved emulsifying ability and further improved the heat stability of the emulsion droplets. Increasing DH from 27 to 35% led to a big decrease in both emulsifying ability and heat stability. The quiescent coalescence stability of WPH emulsions was relatively good up to a DH of 27%. Above DH 27% emulsions become highly unstable. It appears that two mechanisms of instability are at work here. At low DH heat-induced denaturation and aggregation occur. In the DH range of 4-20% heat stability increases as protein globular structure is disrupted. At a DH greater than 27% we see a change from a hydrolysis-induced increase in heat-stability to coalescence instability, with a resultant large increase in emulsion breakdown during heating.     

Correlations between biochemical characteristics and foam-forming and -stabilizing ability of whey and casein hydrolysates

Whey protein and casein were hydrolyzed with 11 commercially available enzymes. Foam properties of 44 samples were measured and were related to biochemical properties of the hydrolysates using statistical data analysis. All casein hydrolysates formed high initial foam levels, whereas whey hydrolysates differed in their foam-forming abilities. Regression analysis using the molecular weight distribution of whey hydrolysates as predictors showed that the hydrolysate fraction containing peptides of 3-5 kDa was most strongly related to foam formation. Foam stability of whey hydrolysates and of most casein hydrolysates was inferior to that of the intact proteins. The foam stability of casein hydrolysate foams was correlated to the molecular weight distribution of the hydrolysates; a high proportion of peptides >7 kDa, composed of both intact casein and high molecular weight peptides, was positively related to foam stability.     

FTIR spectra of whey and casein hydrolysates in relation to their functional properties

Mid-infrared spectra of whey and casein hydrolysates were recorded using Fourier transform infrared (FTIR) spectroscopy. Multivariate data analysis techniques were used to investigate the capacity of FTIR spectra to classify hydrolysates and to study the ability of the spectra to predict bitterness, solubility, emulsifying, and foaming properties of hydrolysates. Principal component analysis revealed that hydrolysates prepared from different protein sources or with different classes of proteolytic enzymes are distinguished effectively on basis of their FTIR spectra. Moreover, multivariate regression analysis showed satisfactory to good prediction of functional parameters; the coefficient of determination (R(2)) varied from 0.60 to 0.92. The accurate prediction of bitterness and emulsion forming ability of hydrolysates by using only one uncomplicated and rapid analytical method has not been reported before. FTIR spectra in combination with multivariate data analysis proved to be valuable in protein hydrolysate fingerprinting and can be used as an alternative for laborious functionality measurements.     

Lipid oxidation in corn oil-in-water emulsions stabilized by casein,whey protein isolate,and soy protein isolate

Proteins can be used to produce cationic oil-in-water emulsion droplets at pH 3.0 that have high oxidative stability. This research investigated differences in the physical properties and oxidative stability of corn oil-in-water emulsions stabilized by casein, whey protein isolate (WPI), or soy protein isolate (SPI) at pH 3.0. Emulsions were prepared with 5% corn oil and 0.2-1.5% protein. Physically stable, monomodal emulsions were prepared with 1.5% casein, 1.0 or 1.5% SPI, and > or =0.5% WPI. The oxidative stability of the different protein-stabilized emulsions was in the order of casein > WPI > SPI as determined by monitoring both lipid hydroperoxide and headspace hexanal formation. The degree of positive charge on the protein-stabilized emulsion droplets was not the only factor involved in the inhibition of lipid oxidation because the charge of the emulsion droplets (WPI > casein > or = SPI) did not parallel oxidative stability. Other potential reasons for differences in oxidative stability of the protein-stabilized emulsions include differences in interfacial film thickness, protein chelating properties, and differences in free radical scavenging amino acids. This research shows that differences can be seen in the oxidative stability of protein-stabilized emulsions; however, further research is needed to determine the mechanisms for these differences.     

Emulsion properties of sunflower (Helianthus annuus) proteins

Emulsions were made with sunflower protein isolate (SI), helianthinin, and sunflower albumins (SFAs). Emulsion formation and stabilization were studied as a function of pH and ionic strength and after heat treatment of the proteins. The emulsions were characterized with respect to average droplet size, surface excess, and the occurrence of coalescence and/or droplet aggregation. Sunflower proteins were shown to form stable emulsions, with the exception of SFAs at neutral and alkaline pH values. Droplet aggregation occurred in emulsions made with SI, helianthinin, and SFAs. Droplet aggregation and subsequent coalescence of emulsions made with SFAs could be prevented at pH 3. Calcium was found to cause droplet aggregation of emulsions made with helianthinin, at neutral and alkaline pH values. Treatments that increase conformational flexibility of the protein molecule improved the emulsion properties of sunflower proteins.     

Influence of calcium, magnesium, or potassium ions on the formation and stability of emulsions prepared using highly hydrolyzed whey proteins

Oil-in-water emulsions (4 wt % soy oil) containing 4 wt % whey protein hydrolysate (WPH) (27% degree of hydrolysis) and different levels of calcium, magnesium, or potassium chloride were prepared in a two-stage homogenizer. Other emulsions containing 4 wt % WPH but including 0.35 wt % hydroxylated lecithin and different levels of the above minerals were similarly prepared. The formation and stability of these emulsions were determined by measuring oil droplet size distributions using laser light scattering and by confocal scanning laser microscopy and a gravity creaming test. Both lecithin-free and lecithin-containing emulsions showed no change in droplet size distributions with increasing concentration of potassium in the range 0-37.5 mM. In contrast, the diameter of emulsion droplets increased with increasing calcium or magnesium concentration >12.5 mM. Emulsions containing hydroxylated lecithin were more sensitive to the addition of calcium or magnesium than the lecithin-free emulsions. Storage of emulsions at 20 degrees C for 24 h further increased the diameter of droplets and resulted in extensive creaming in emulsions containing >25 mM calcium or magnesium. It appears that both flocculation and coalescence processes were involved in the destabilization of emulsions induced by the addition of divalent cations.     

Rheology and stability of acidified food emulsions treated with high pressure

The stability and rheology of acidified model oil-in-water emulsions (pH 3.6 +/- 0.1) were evaluated before and after high-pressure treatments. Varying concentrations of canola oil (0-50% w/w), whey protein isolate, polysorbate 60, soy lecithin (0.1-1.5% w/w each), and xanthan (0.0-0.2% w/w) were chosen. Exposure to high pressures (up to 800 MPa for 5 min at 30 degrees C) did not significantly affect the equivalent surface mean diameter D[3,2], flow behavior, and viscoelasticity of the whey protein isolate and polysorbate 60-stabilized emulsions. Pressure treatments had negligible effects on emulsion stability in these systems, except when xanthan (0.2% w/w) was present in which pressure improved the stability of polysorbate 60-stabilized emulsions. Soy lecithin-stabilized emulsions had larger mean particles sizes and lower emulsion volume indices than the others, indicating potential instability, and application of pressure further destabilized these emulsions.     

Influence of polysaccharides on the rate of coalescence in oil-in-water emulsions formed with highly hydrolyzed whey proteins

The objective of this study was to examine the effects of added xanthan gum, guar gum, or kappa-carrageenan on the formation and properties of emulsions (4 wt % corn oil) formed with an extensively hydrolyzed commercial whey protein (WPH) product under a range of conditions. The rate of coalescence was calculated on the basis of the changes in the droplet size of emulsions during storage of the emulsions at 20 degrees C. Compared with the emulsion made without the addition of polysaccharides, the rate of creaming and coalescence in emulsions containing xanthan gum, guar gum, or kappa-carrageenan was markedly enhanced with increasing concentration of polysaccharides during storage for up to 7 days. At a given concentration, the rate of coalescence was highest in the emulsions containing guar gum, whereas it was lowest in the emulsions containing kappa-carrageenan. All emulsions containing xanthan gum, guar gum, or kappa-carrageenan showed flocculation of oil droplets by a depletion mechanism. This flocculation was considered to enhance the coalescence of oil droplets. The different rates of coalescence could be explained on the basis of the strength of the depletion potential, which was dependent on the molecular weight and the radius of gyration of the polysaccharides.     

甘薯热变性蛋白限制性酶解产物的乳化特性研究

采用5种酶(Alcalase 2.4L, As1.398, Neutrase, Pepsin, Trypsin)对甘薯热变性蛋白(SPHP)进行限制性酶解。将各酶解产物离心后分别取上清和沉淀测定和观察其乳化液的乳化颗粒平均粒径(D_4,3)、乳化活性指数(EAI)、乳化稳定性指数(ESI)、乳化液的微观结构和表观黏度。结果显示:酶解产物上清和沉淀中蛋白的溶解度均有增加,但沉淀增加的幅度小于上清。SPHP的D_4,3是71.96μm,而酶解产物上清和沉淀乳化液的D_4,3均减小,且上清的D_4,3小于沉淀的。在5种酶解产物中,Pepsin酶解物上清的D_4,3最小,为14.94μm。SPHP酶解后上清的乳化颗粒大小较为均一,且沉淀的乳化颗粒酶解前后变化不大。SPHP的EAI为11.21m2/g,酶解产物上清和沉淀的EAI均有显著提高(P<0.05),其中Pepsin酶解物上清的EAI最高为70.32m2/g。此外,酶解产物上清和沉淀乳化液的ESI增大。与沉淀相比,5种酶解产物的上清具有较低的表观黏度,且酶解产物上清和沉淀的乳化液均呈剪切变稀的非牛顿流体特性。     

Preparation and characterization of papain-modified sesame (Sesamum indicum L.) protein isolates

Defatted sesame meal ( approximately 40-50% protein content) is very important as a protein source for human consumption due to the presence of sulfur-containing amino acids, mainly methionine. Sesame protein isolate (SPI) is produced from dehulled, defatted sesame meal and used as a starting material to produce protein hydrolysate by papain. Protein solubility at different pH values, emulsifying properties in terms of emulsion activity index (EAI) and emulsion stability index (ESI), foaming properties in terms of foam capacity (FC) and foam stability (FS), and molecular weight distribution of the SPI hydrolysates were investigated. Within 10 min of hydrolysis, the maximum cleavage of peptide bonds occurred as observed from the degree of hydrolysis. Protein hydrolysates have better functional properties than the original SPI. Significant increase in protein solubility, EAI, and ESI were observed. The greatest increase in solubility was observed between pH 5.0 and 7.0. The molecular weight of the hydrolysates was also reduced significantly during hydrolysis. These improved functional properties of different protein hydrolysates would make them useful products, especially in the food, pharmaceutical, and related industries.     

beta-lactoglobulin hydrolysis. 1. Peptide composition and functional properties of hydrolysates obtained by the action of plasmin, trypsin, and Staphylococcus aureus V8 protease.

beta-Lactoglobulin (betaLg) was subjected to limited hydrolysis by trypsin, plasmin, and endoproteinase from Staphylococcus aureus V8 (S.aur.V8) to degrees of hydrolysis (DH) of 1, 2, and 4%. The several hydrolysates had different peptide compositions (determined by reversed-phase HPLC and gel-permeation chromatography [GPC]). GPC under nondenaturing, denaturing, and denaturing plus reducing conditions showed that the peptides formed were linked by hydrophobic interactions or by disulfide bonds or were not linked at all. At very low protein concentration, some differences in emulsion-forming properties were observed: only the plasmin hydrolysates could form emulsions with a uniform particle-size distribution. The emulsions formed with S.aur.V8 hydrolysates had poor emulsion-stabilizing properties. Some hydrolysates showed increased foam-forming properties in comparison with the intact protein. All foams formed were stable. Overall, the plasmin hydrolysate (DH4) contained relatively much larger molecules and/or hydrophobic molecules. Many molecules were disulfide-linked peptides. This hydrolysate also had the best functional properties.     

Preparation and characterization of beta-carotene nanodispersions prepared by solvent displacement technique

This work demonstrated the preparation of protein-stabilized beta-carotene nanodispersions using the solvent displacement technique. The emulsifying performance of sodium caseinate (SC), whey protein concentrate (WPC), whey protein isolate (WPI), and a whey protein hydrolysate (WPH, 18% degree of hydrolysis) was compared in terms of particle size and zeta-potential of the nanodispersions. SC-stabilized nanodispersions exhibited a bimodal particle size distribution: large particles (stabilized by casein micelles) with a mean particle size of 171 nm and small particles (stabilized by casein submicelles) of 13 nm. This was confirmed with transmission electron microscopy analysis. Most of the beta-carotene precipitated (87.6%) was stabilized in the small particles. On the other hand, the nanodispersions stabilized by the whey proteins were polydispersed with larger mean particle sizes. The mean particle size of WPC and WPI was 1730 and 201 nm, respectively. The SC-stabilized nanodispersion was expected to be more stable as indicated by its higher absolute zeta-potential value (-31 mV) compared to that of WPC (-15 mV) and WPI (-16 mV). Partially hydrolyzed whey protein possessed improved emulsifying properties as shown by WPH-stabilized samples. It was interesting to note that increasing the SC concentration from 0.05 to 0.5 wt % increased the particle size of beta-carotene stabilized by casein micelles, while the reverse was true for those stabilized by SC submicelles. Microfluidization at 100 MPa of SC solution dissociated the casein micelles, resulting in a decrease in mean particle size of the casein micelle-stabilized particles when the SC solution was used to prepare nanodispersions. The results from this work showed that protein-stabilized beta-carotene nanodispersions could be prepared using the solvent displacement technique.     

Improved storage stability of model infant formula by whey peptides fractions

The purpose of this study was to evaluate the shelf-life stability (6 months) of model infant formula with whey protein hydrolysates or peptidic fractions as carrageenan replacers. Whey protein hydrolysates were prepared with trypsin and followed by ultrafiltration of the hydrolyzed mixture, and peptidic fractions were isolated from the ultrafiltered tryptic hydrolysate by anion- or cation-exchange chromatography. The stability of the model infant formula was evaluated using a stratification method based on fat content differences between the top and bottom strata of the samples. With protein hydrolysate-based formulations, the creaming rate of the fat in the product was slightly higher than in the standard formulation (with carrageenan), which is indicative of lower storage stability. The addition of cationic fractions to model infant formula also resulted in lower product stability, whereas the fat creaming rate was retarded in anionic fraction based formulations. The physicochemical characteristics of certain peptides combined with the reported high emulsifying properties of peptidic sequences found within these fractions may account for their ability to act as carrageenan replacers.     

Microencapsulating properties of sodium caseinate

Emulsions were prepared with 5% (w/v) solutions of sodium caseinate (Na Cas) and soy oil at oil/protein ratios of 0.25-3.0 by homogenization at 10--50 MPa. Emulsions were spray-dried to yield powders with 20--75% oil (w/w). Emulsion oil droplet size and interfacial protein load were determined. Microencapsulation efficiency (ME), redispersion properties, and structure of the powders were analyzed. The size of emulsion oil droplets decreased with increasing homogenization pressure but was not influenced by oil/protein ratio. Emulsion protein load values were highest at low oil/protein ratios. ME of the dried emulsions was not affected by homogenization pressure but decreased from 89.2 to 18.8% when the oil/protein ratio was increased from 0.25 to 3.0, respectively. Mean particle sizes of reconstituted dried emulsions were greater than those of the original emulsions, particularly at high oil/protein ratios (>1.0), suggesting destabilization of high-oil emulsions during the spray-drying process.     

Impact of whey protein emulsifiers on the oxidative stability of salmon oil-in-water emulsions

To obtain a better understanding of how the interfacial region of emulsion droplets influences lipid oxidation, the oxidative stability of salmon oil-in-water emulsions stabilized by whey protein isolate (WPI), sweet whey (SW), beta-lactoglobulin (beta-Lg), or alpha-lactalbumin (alpha-La) was evaluated. Studies on the influence of pH on lipid oxidation in WPI-stabilized emulsions showed that formation of lipid hydroperoxides and headspace propanal was much lower at pH values below the protein's isoelectric point (pI), at which the emulsion droplets were positively charged, compared to that at pH values above the pI, at which the emulsion droplets were negatively charged. This effect was likely due to the ability of positively charged emulsion droplets to repel cationic iron. In a comparison of lipid oxidation rates of WPI-, SW-, beta-Lg-, and alpha-La-stabilized emulsions at pH 3, the oxidative stability was in the order of beta-Lg > or = SW > alpha-La > or = WPI. The result indicated that it was possible to engineer emulsions with greater oxidative stability by using proteins as emulsifier, thereby reducing or eliminating the need for exogenous food antioxidants.     

Use of caseinophosphopeptides as natural antioxidants in oil-in-water emulsions

Chelators are valuable ingredients used to improve the oxidative stability of food emulsions. Caseins and casein peptides have phosphoseryl residues capable of binding transition metals. Thus, the ability of enriched caseinophosphopeptides to inhibit lipid oxidation in corn oil-in-water emulsions was investigated. Enriched caseinophosphopeptides (25 microM) inhibited the formation of lipid oxidation at both pH 3.0 and 7.0 as determined by lipid hydroperoxides and hexanal. Calcium (0-100 mM) had no influence on the antioxidant activity of the enriched caseinophosphopeptides. Casein hydrolysates were more effective inhibitors of lipid oxidation than the enriched caseinophosphopeptides at equal phosphorus content. Thus, antioxidant properties might not be uniquely attributed to chelating metals by phosphoseryl residues but also by scavenging free radicals. Overall, the observed antioxidant activity of casein hydrolysates means they could be utilized to decrease oxidative rancidity in foods.     

Stabilization of oil-in-water emulsions by beta-lactoglobulin-polyethylene glycol conjugates

The disulfide bonds of beta-lactoglobulin (beta-lg) were modified by oxidative sulfitolysis to generate beta-lgSO(3). The native protein (beta-lg) and the modified protein (beta-lgSO(3)) were conjugated to activated polyethylene glycol (PEG) to generate beta-lgPEG and beta-lgSO(3)PEG, respectively. Oil-in-water (o/w) emulsions containing 1% beta-lg or beta-lg conjugates were prepared at pH 2.8, 5.0, and 7.0. Emulsion droplet diameters and zeta potentials were measured. For the same emulsifier, emulsion droplet diameters decreased when emulsion pH increased. Zeta potentials of emulsion droplets increased with pH for beta-lg and beta-lgSO(3). Zeta potentials of beta-lgPEG and beta-lgSO(3)PEG approached zero, suggesting that the protein molecule was covered by PEG chains. Accelerated and 7-day storage stabilities at 21 degrees C of the emulsions were monitored. The emulsifying activity index (EAI) of beta-lgPEG was not significantly different from the EAI of beta-lg. The EAI of beta-lg was enhanced following sulfitolysis of beta-lactoglobulin. The emulsifying activity increased more when the oxidatively modified protein was conjugated to polyethylene glycol. Emulsions made with beta-lgSO(3)PEG were more stable than emulsions made with beta-lg, beta-lgPEG, or beta-lgSO(3) under accelerated stability study and for 7 days at 21 degrees C. The stability of o/w emulsions stabilized with beta-lgSO(3)PEG increased because individual droplets were better protected, against protein bridging or coalescence, by the thick adsorbed protein-PEG layer.     

Arabinoxylan-lipid-based edible films and coatings. 3. Influence of drying temperature on film structure and functional properties

This work is a contribution to better knowledge of the influence of the structure of films obtained from emulsions based on arabinoxylans, hydrogenated palm kernel oil, and emulsifiers on their functional properties. The sucrose esters (emulsifiers) have a great effect on the stabilization of the emulsified film structure containing arabinoxylans and hydrogenated palm kernel oil. The structure and stability of the emulsion during drying strongly affect barrier and mechanical properties of films. The higher are creaming and coalescence phenomena in films, the lower is the water vapor permeability. Emulsion destabilization is favored by high drying temperature and tends to give films having a "bilayer-like" structure, which tends to improve the functional properties of arabinoxylans-based edible films.     

Preparation of antioxidant enzymatic hydrolysates from alpha-lactalbumin and beta-lactoglobulin. Identification of active peptides by HPLC-MS/MS

We have investigated the antioxidant activity of hydrolysates from whey proteins bovine alpha-lactalbumin (alpha-La) and beta-lactoglobulin A (beta-Lg A) by commercial proteases (pepsin, trypsin, chymotrypsin, thermolysin, and Corolase PP). Corolase PP was the most appropriate enzyme to obtain antioxidant hydrolysates from alpha-La and beta-Lg A (ORAC-FL values of 2.315 and 2.151 micromol of Trolox equivalent/mg of protein, respectively). A total of 42 peptide fragments were identified by HPLC-MS/MS in the beta-Lg A hydrolysate by Corolase PP. One of the sequences (Trp-Tyr-Ser-Leu-Ala-Met-Ala-Ala-Ser-Asp-Ile) possessed radical scavenging (ORAC-FL value of 2.621 micromol of Trolox equivalent/micromol of peptide) higher than that of butylated hydroxyanisole (BHA). Our results suggest that whey protein hydrolysates could be suitable as natural ingredients in enhancing antioxidant properties of functional foods and in preventing oxidation reaction in food processing.     

Lipid oxidation in a menhaden oil-in-water emulsion stabilized by sodium caseinate cross-linked with transglutaminase

Transglutaminase-catalyzed cross-linking of interfacial proteins in oil-in-water has been shown to influence physical stability, but little is known about how this reaction impacts lipid oxidation. Therefore, this study evaluated the influence of transglutaminase-induced interfacial protein cross-linking on the oxidative stability of casein-stabilized menhaden oil-in-water emulsions. Interfacial casein in menhaden oil-in-water emulsions cross-linked by transglutaminase (pH 7.0) produced a cohesive interfacial protein layer that could not be removed from the emulsion droplet by Tween 20. Although transglutaminase cross-linked the interfacial casein, these emulsions did not show increased oxidative stability when compared to untreated emulsions as determined by measurement of lipid hydroperoxides and thiobarbituric acid reactive substances. These results indicate that increasing the cohesiveness of proteins at the interface of oil-in-water emulsions does not inhibit lipid oxidation. This could be due to the ability of prooxidative species such as iron to diffuse through the cross-linked protein layer where it could promote the decomposition of lipid hydroperoxides into free radicals that could oxidize unsaturated fatty acids in the emulsion droplet core.