Relationship between the microstructure and the mechanical and barrier properties of whey protein films

This work was focused on the relationship between the microstructure and the mechanical and barrier properties of whey protein isolate (WPI) films. Sorbitol (S) and glycerol (G) were used as plasticizers and the pH was varied between 7 and 9. The films were cast from heated aqueous solutions and dried in a climate room at 23 degrees C and 50% relative humidity for 16 h. The microstructure of the films was found to be dependent on the concentration, the plasticizers, and the pH. When the concentration increased, a more aggregated structure was formed, with a denser protein network and larger pores. This resulted in increased water vapor permeability (WVP) and decreased oxygen permeability (OP). When G was used as a plasticizer instead of S, the microstructure was different, and the moisture content and WVP approximately doubled. When the pH increased from 7 to 9, a denser protein structure was formed, the strain at break increased, and the OP decreased.     

Aging of whey protein films and the effect on mechanical and barrier properties

This work focuses on the aging of whey protein isolate (WPI) films plasticized with glycerol (G) and sorbitol (S). The films were cast from heated aqueous solutions at pH 7 and dried at 23 degrees C and 50% relative humidity (RH) for 16 h. They were stored in a climate room (23 degrees C, 50% RH) for 120 days, and the film properties were measured at regular intervals. The moisture content (MC) of the WPI/G films decreased from 22% (2 days) to 15% (45 days) and was thereafter constant at 15% (up to 120 days). This affected the mechanical properties and caused an increased stress at break (from 2.7 to 8.3 MPa), a decreased strain at break (from 33 to 4%), and an increased glass transition temperature (T(g)) (from -56 to -45 degrees C). The barrier properties were, however, unaffected, with constant water vapor permeability and a uniform film thickness. The MC of the WPI/S films was constant at approximately 9%, which gave no change in film properties.     

Barrier and mechanical properties of milk protein-based edible films containing nutraceuticals

Calcium caseinate (CC) and whey protein isolate (WPI) films were prepared to contain 5 or 10% Gluconal Cal (GC), a mixture of calcium lactate and gluconate, or 0.1 or 0.2% alpha-tocopheryl acetate (VE), respectively. The pH and viscosity of film-forming solutions and the water vapor permeability and tensile property of the films were determined using standard procedures. CC and WPI films have the capabilities to carry high concentration of GC or VE, but some of the film functionality might be compromised. Adding VE to CC and WPI films increased film elongation at break, whereas incorporating 0.2% VE decreased WVP of CC films and tensile strength of both CC and WPI films. Incorporation of GC reduced the tensile strength of CC films (P < 0.05), with 10% GC decreasing both elongation at break and WVP (P < 0.05). These types of films may be used for wrapping or coating to enhance the nutritional value of foods. The concentration of GC and VE added to the films must be carefully selected to meet required water barrier and mechanical properties of the films depending on their specific applications.     

Plasticizer effect on oxygen permeability of beta-lactoglobulin films

Plasticizer effect on oxygen permeability (OP) of beta-lactoglobulin (beta-Lg) films was studied. Propylene glycol (PG), glycerol (Gly), sorbitol (Sor), sucrose (Suc), and polyethylene glycol at MW 200 and 400 (PEG 200 and PEG 400, respectively) were studied due to their differences in composition, shape, and size. Suc-plasticized beta-Lg films gave the best oxygen barrier (OP < 0.05 cm3 x microm/m2 x day x kPa). Gly- and PG-plasticized films had similar OP values, and both had higher OP than Sor-plasticized films. PEG 200- and PEG 400-plasticized films were the poorest oxygen barriers. Empirical equations including plasticizer efficiencies for OP were employed to elucidate the relationships between OP of plasticized beta-Lg films and plasticizer type and content. Plasticizer efficiency ratios between mechanical and OP properties of beta-Lg films show the relative efficiency of plasticizers in modifying mechanical and OP properties. A large ratio is desirable.     

Lipid particle size effect on water vapor permeability and mechanical properties of whey protein/beeswax emulsion films

Lipid particle size effects on water vapor permeability (WVP) and mechanical properties of whey protein isolate (WPI)/beeswax (BW) emulsion films were investigated. Emulsion films containing 20 and 60% BW (dry basis) and mean lipid particle sizes ranging from 0.5 to 2.0 microm were prepared. BW particle size effects on WVP and mechanical properties were observed only in films containing 60% BW. WVP of these films decreased as lipid particle size decreased. As drying temperature increased, film WVPs decreased significantly. Meanwhile, tensile strength and elongation increased as BW particle size decreased. However, for 20% BW emulsion films, properties were not affected by lipid particle size. Results suggest that increased protein-lipid interactions at the BW particle interfaces, as particle size decreased and resulting interfacial area increased, result in stronger films with lower WVPs. Observing this effect depends on a large lipid content within the protein matrix. At low lipid content, the effect of interactions at the protein-lipid interfaces is not observed, due to the presence of large protein-matrix regions of the film without lipid, which are not influenced by protein-lipid interactions.     

Drying temperature effect on water vapor permeability and mechanical properties of whey protein-lipid emulsion films

The water vapor permeability (WVP) and mechanical properties of whey protein isolate (WPI) and WPI-lipid emulsion films dried at different conditions were investigated. As drying temperature increased, WVPs decreased significantly. Significantly lower WVP was observed for emulsion films compared to WPI films. WPI-Beeswax (BW) and WPI-anhydrous milkfat fraction emulsion films dried at 80 degrees C and 40% RH gave the lowest WVP compared to 25 degrees C, 40% RH and 40 degrees C, 40% RH. A large drop in WVP of WPI-BW emulsion films was observed at 20% BW content. The decrease in WVP for emulsion films as drying temperature increased could be due to change in the lipid crystalline morphology and/or lipid distribution within the matrix. Mechanical properties of WPI and WPI-lipid emulsion films, on the other hand, were not modified by drying conditions.     

Enzymatic hydrolysis of heat-induced aggregates of whey protein isolate

The effects of heat-induced denaturation and subsequent aggregation of whey protein isolate (WPI) solutions on the rate of enzymatic hydrolysis was investigated. Both heated (60 °C, 15 min; 65 °C, 5 and 15 min; 70 °C, 5 and 15 min, 75 °C, 5 and 15 min; 80 °C, 10 min) and unheated WPI solutions (100 g L(-1) protein) were incubated with a commercial proteolytic enzyme preparation, Corolase PP, until they reached a target degree of hydrolysis (DH) of 5%. WPI solutions on heating were characterized by large aggregate formation, higher viscosity, and surface hydrophobicity and hydrolyzed more rapidly (P < 0.001) than the unheated. The whey proteins exhibited differences in their susceptibility to hydrolysis. Both viscosity and surface hydrophobicity along with insolubility declined as hydrolysis progressed. However, microstructural changes observed by light and confocal laser scanning microscopy (CLSM) provided insights to suggest that aggregate size and porosity may be complementary to denaturation in promoting faster enzymatic hydrolysis. This could be clearly observed in the course of aggregate disintegration, gel network breakdown, and improved solution clarification.     

Ascorbic acid-containing whey protein film coatings for control of oxidation

A formulation for the whey protein isolate film or coating incorporating ascorbic acid (AA-WPI film or coating) was developed. Tensile and oxygen-barrier properties of the AA-WPI film were measured. Antioxidant effects of the AA-WPI coating on roasted peanuts were studied by comparing the values of peroxide (PO), thiobarbituric acid reactive substance (TBARS), and free-radical-scavenging activity, determined with noncoated peanuts and peanuts coated with WPI with and without ascorbic acid during storage at 21% relative humidity (RH) and 23, 35, and 50 degrees C. The incorporation of AA reduced elongation of WPI films. The oxygen-barrier property of the WPI film was significantly improved by incorporation of AA. The AA-WPI coating retarded lipid oxidation in peanuts significantly at 23, 35, and 50 degrees C. The AA-WPI coated peanuts were more red than noncoated peanuts at all storage temperatures.     

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.     

Effects of pH and the gel state on the mechanical properties, moisture contents, and glass transition temperatures of whey protein films.

The mechanical properties, moisture contents (MC), and glass transition temperature (T(g)) of whey protein isolate (WPI) films were studied at various pH values using sorbitol (S) as a plasticizer. The films were cast from heated aqueous solutions and dried in a climate chamber at 23 degrees C and 50% relative humidity (RH) for 16 h. The critical gel concentrations (c(g)) for the cooled aqueous solutions were found to be 11.7, 12.1, and 11.3% (w/w) WPI for pH 7, 8, and 9, respectively. The cooling rate influenced the c(g), in that a lower amount of WPI was needed for gelation when a slower cooling rate was applied. Both cooling rates used in this study showed a maximum in the c(g) at pH 8. The influence of the polymer network on the film properties was elucidated by varying the concentration of WPI over and under the c(g). Strain at break (epsilon(b)) showed a maximum at the c(g) for all pH values, thus implying that the most favorable structure regarding the ability of the films to stretch is formed at this concentration. Young's modulus (E) and stress at break (sigma(b)) showed a maximum at c(g) for pH 7 and 8. The MC and epsilon(b) increased when pH increased from 7 to 9, whereas T(g) decreased. Hence, T(g) values were -17, -18, and -21 degrees C for pH 7, 8, and 9, respectively. E and sigma(b) decreased and epsilon(b) and thickness increased when the surrounding RH increased. The thickness of the WPI films also increased with the concentration of WPI.     

Effects of alpha-tocopherol, beta-carotene, and soy isoflavones on lipid oxidation of structured lipid-based emulsions

Structured lipids (SLs) are triacylglycerols that have been modified to change the fatty acid composition and/or positional distribution in the glycerol backbone by chemically and/or enzymatically catalyzed reactions and/or genetic engineering. Ten percent oil-in-water emulsions were formulated with a canola oil/caprylic acid SL and stabilized with 0.5% whey protein isolate (WPI) or sucrose fatty acid ester (SFE). The effects of alpha-tocopherol, beta-carotene, genistein, and daidzein (added at 0.02 wt % of oil) on lipid oxidation were evaluated over a 15-day period in emulsion samples. Significantly (p < 0.05) less total oxidation (calculated from peroxide value and anisidine value measurements) occurred in the WPI emulsions compared to their SFE counterparts. In this study, alpha-tocopherol, beta-carotene, and both soy isoflavones exhibited prooxidant activities in SFE emulsions. Because of their ability to exhibit prooxidant activity under certain conditions, manufacturers must experiment with these compounds before adding them to SL-based products as functional ingredients.     

Structural changes imposed on whey proteins by UV irradiation in a continuous UV light reactor

The objective of this study was to investigate the structural changes of whey proteins during exposure in a continuous-flow UV reactor. Varying UV irradiation dosages were obtained by controlling the flow rate and the mixing speed. Whey protein isolate (WPI) solutions at concentrations of 1% and 5% (w/v) were circulated at flow rates ranging from 30 to 800 mL·min(-1), and changes in physicochemical properties of the proteins were investigated. Intrinsic fluorescence spectra and surface hydrophobicity measurements suggested changes in the tertiary structure of the proteins with UV exposure. The UV treatment also increased the concentration of total and accessible thiol groups in 1% WPI solutions, while no change was measured in 5% WPI solutions. Size-exclusion chromatography demonstrated the formation of UV-induced aggregates and oxidation products (N-formylkynurenine and dityrosine) of aromatic amino acids. Furthermore, the UV-induced changes in protein conformation increased the susceptibility of whey proteins to pepsin hydrolysis.     

Effects of ionic strength on the enzymatic hydrolysis of diluted and concentrated whey protein isolate

To identify the parameters that affect enzymatic hydrolysis at high substrate concentrations, whey protein isolate (1-30% w/v) was hydrolyzed by Alcalase and Neutrase at constant enzyme-to-substrate ratio. No changes were observed in the solubility and the aggregation state of the proteins. With increasing concentration, both the hydrolysis rate and the final DH decreased, from 0.14 to 0.015 s(-1) and from 24 to 15%, respectively. The presence of 0.5 M NaCl decreased the rate of hydrolysis for low concentrations (to 0.018 s(-1) for 1% WPI), resulting in similar rates of hydrolysis for all substrate concentrations. The conductivity increase (by increasing the protein concentration, or by addition of NaCl) has significant effects on the hydrolysis kinetics, but the reason for this is not yet well understood. The results show the importance of conductivity as a factor that influences the kinetics of the hydrolysis, as well as the composition of the hydrolysates.     

Mechanical and barrier properties of cross-linked soy and whey protein based films

Sterilized biofilms based on soy protein isolate (SPI, S system) and a 1:1 mixture of SPI and whey protein isolate (WPI, SW system) were achieved through the formation of cross-links by means of gamma-irradiation combined with thermal treatments. The effect of the incorporation of carboxymethylcellulose (CMC) and poly(vinyl alcohol) was also examined. gamma-Irradiation combined with thermal treatment improved significantly the mechanical properties, namely, puncture strength and puncture deformation, for all types of films. Irradiated formulations that contain CMC behave more similarly as elastomers. CMC showed also significant improvements of the barrier properties, namely, water vapor permeability, for irradiated films of the S system and for non-irradiated films of the SW system.     

Storage stability of ascorbic acid incorporated in edible whey protein films

The stability of ascorbic acid (AA) incorporated in whey protein isolate (WPI) film and the related color changes during storage were studied. No significant loss of AA content was found in any films prepared from pH 2.0 casting solution stored at 30% relative humidity (RH) and 22 °C over 84 days. Total visible color difference (ΔE*(ab)) of all films slowly increased over storage time. The ΔE*(ab) values of pH 3.5 films were significantly higher than those of pH 2.0 films. The stability of AA-WPI films was found to be mainly affected by the pH of the film-forming solution and storage temperature. Oxidative degradation of AA-WPI films followed Arrhenius behavior. Reduction of the casting solution pH to below the pK(a1) (4.04 at 25 °C) of AA effectively maintained AA-WPI storage stability by greatly reducing oxidative degradation, whereas anaerobic and nonenzymatic browning were insignificant. The half-life of pH 2.0 AA-WPI film at 30% RH and 22 °C was 520 days.     

Dynamic interfacial rheology as a tool for the characterization of whey protein isolates gelation at the oil-water interface.

Heat-induced interfacial aggregation of a whey protein isolate (WPI), previously adsorbed at the oil-water interface, was studied by interfacial dynamic characteristics coupled with microscopic observation and image analysis of the drop after heat treatment. The experiments were carried out at temperatures ranging from 20 to 80 degrees C with different thermal regimes. During the heating period, competition exists between the effect of temperature on the film fluidity and the increase in mechanical properties associated with the interfacial gelation process. During the isothermal treatment, the surface dilational modulus, E, increases, and the phase angle, delta, decreases with time to a plateau value. The frequency dependence of E and delta is characteristic of viscoelastic films with increasing delta and decreasing E at lower frequencies. The effects of heat treatment depend on the conditions at which the gelation process takes place. Microscopic observation of gelled films gives complementary information on the effect of heat treatment on WPI adsorbed films.     

Gamma-irradiation influence on the structure and properties of calcium caseinate-whey protein isolate based films. Part 1. Radiation effect on the structure of proteins gels and films

Brookfield viscosimetry, Fourier transform infrared spectroscopy, transmission electron microscopy (TEM), and measurements of the texture strength of gels formed with CaCl2 and the mechanical and barrier properties of the film were applied in studies of gel formation and structural and mechanical properties of gels and films prepared using calcium caseinate (CC)-whey protein isolate (WPI)-glycerol (1:1:1), control, and irradiated with 60Co gamma rays using a 32 kGy dose. The irradiated gels have appeared to be more "fine-stranded" as compared to the more "particulate" control gels and lead to the formation of more rigid films with improved mechanical strength and barrier properties. This results from cross-linking and the modification of protein conformations were induced by irradiation, in particular the increase in the beta-sheet and beta-strand contents. Structural modifications taking place in CC-WPI composition are related to modifications taking place separately in CC and WPI. Improvement of the properties of the films after irradiation corresponds to the increased density of the cross-linked material because no change in the porosity of the films was observed by TEM.     

Fractionation of whey protein isolate with supercritical carbon dioxide to produce enriched α-lactalbumin and β-lactoglobulin food ingredients

An environmentally friendly protein fractionation process using supercritical carbon dioxide (SCO(2)) as an acid was developed to produce enriched α-lactalbumin (α-LA) and β-lactoglobulin (β-LG) fractions from whey protein isolate solutions containing from 2 to 10% WPI. This study investigated the effects of pH, temperature, WPI concentration, and residence time on the precipitation kinetics and recovery yields of individual whey proteins and the relative enrichment and composition of both protein fractions. At 5.5-34 MPa and 60-65 °C, solubilized SCO(2) decreased solution pH and induced the formation and precipitation of α-LA aggregates. Gel electrophoresis and HPLC of the enriched fractions demonstrated the production of ≥ 60% pure α-LA, and ≥ 70% pure β-LG, under various operating conditions, from WPI containing ～57% β-LG and 21% α-LA. The enriched fractions are ready-to-use food ingredients with neutral pH, untainted by acids and contaminants.     

Mechanical and structural properties of milk protein edible films cross-linked by heating and gamma-irradiation

The mechanical properties of cross-linked edible films based on calcium caseinate and two type of whey proteins (commercial and isolate) were investigated. Cross-linking of the proteins was carried out using thermal and radiative treatments. Size-exclusion chromatography performed on the cross-linked proteins showed that gamma-irradiation increased the molecular weight of calcium caseinate, while it changed little for the whey proteins. However, heating of the whey protein solution induced cross-linking. For both cross-linked proteins, the molecular weight distribution was >/=2 x 10(3) kDa. Combined thermal and radiative treatments were applied to protein formulations with various ratios of calcium caseinate and whey proteins. Whey protein isolate could replace up to 50% of calcium caseinate without decreasing the puncture strength of the films. Films based on commercial whey protein and calcium caseinate were weaker than those containing whey protein isolate. Electron microscopy showed that the mechanical characteristics of these films are closely related to their microstructures.     

Oxygen permeability of films made from CO2-precipitated casein and modified casein

Oxygen permeabilities (OP) of CO(2)-casein (CO(2)CN), calcium caseinate (CaCN), and acylated casein (AcCN) films were determined as functions of % relative humidity (% RH), temperature, and plasticizer type. Tensile properties and water vapor permeabilities (WVP) were also measured. Plasticizers were glycerol (GLY) or a 3:1 ratio of GLY:poly(propylene glycol) (PPG), a hydrophobic plasticizer. OP of the CO(2)CN:GLY film was almost twice that of films containing either plasticizer at 35% RH, but its OP approached that of the other films at 70% RH. OP and WVP of films plasticized with GLY were greater than that for films plasticized with PPG. Plasticizer type had little impact on the tensile strength of CO(2)CN films while tensile strength of CaCN-GLY:PPG (3:1) films approximately doubled. Results show that structural dissimilarities in the films contribute to differences in OP only under conditions of low RH where the plasticizing effects of water are not significant.