Interfacial dilatational elasticity and viscosity of beta-lactoglobulin at air-water interface using pulsating bubble tensiometry

The ability of proteins to provide stability in foams is greatly influenced by their interfacial dilatational rheological properties. Surface tension response of a pulsatingbubble with an adsorbed layer of beta-lactoglobulin was measured for different frequencies and protein concentrations using a pulsating bubble tensiometer. A methodology, accounting for adsorption/desorption as well as variation of surface concentration due to expansion/contraction, was developed for the evaluation of surface dilatational elasticity and viscosity at different frequencies from these measurements. The adsorption rate constants were inferred from the surface pressure dynamics of protein adsorption using a Langmuir minitrough. The desorption rates were shown to be negligible for beta-lactoglobulin from the surface pressure response of a spread monolayer when subjected to compression in a Langmuir minitrough. The proposed model was employed to infer the interfacial dilatational viscosity and elasticity of an adsorbed beta-lactoglobulin layer at the air-water interface from experimental pulsating bubble data for protein concentrations in the range of 0.01-0.5 wt % at pH 7. As expected, the interfacial dilatational rheological properties were found to be higher at higher protein concentrations, this effect being less pronounced for dilatational elasticity. Heating at 80 degrees C for 30 min was found to result in higher interfacial dilatational viscosity and lower interfacial dilatational elasticity though this difference was within experimental error. The traditional approach for the inference of interfacial dilatational rheological properties is found to overpredict the interfacial dilatational elasticity whereas the viscosity values do not differ significantly from those obtained using the current analysis.     

Structure and mechanical properties of a polyglycerol ester at the air-water surface

We studied the structure and mechanical properties of surface films resulting from the adsorption of a dispersed L beta phase at the air-water interface. This L beta phase corresponds to multilamellar vesicles and is formed by a commercial polyglycerol fatty acid ester (PGE) in aqueous solution at temperatures below the main chain-melting temperature (Tm=58 degrees C). We measured the adsorption kinetics using the pendant drop technique and mechanical properties of PGE films using oscillatory surface shear and dilatational rheometric methods. Though the adsorption kinetics are very slow, we show that the L beta phase of PGE is surface-active and forms viscoelastic films at the air-water surface after sufficiently long adsorption times. The rheological response functions to shear and dilatational deformation are reminiscent of those of temporary networks, indicating an intermolecular connectivity at the surface. This temporary network is probably created by hydrophobic interactions of alkyl chains. We obtained more detailed information about the properties of this network by comparing the rheological signature of an adsorbed PGE film (unknown structure) with a solvent-spread monolayer (known structure). We characterized the structural features of spread PGE films by recording the Langmuir isotherm and Brewster angle micrographs (BAM).We show that the rheological responses of the adsorbed film and the solvent-spread monolayer are very close to each other, indicating a structural similarity. From this study, we conclude that a dispersed L beta phase of PGE is able to adsorb at the air-water surface at T相似文献   

Modulating surface rheology by electrostatic protein/polysaccharide interactions

There is a large interest in mixed protein/polysaccharide layers at air-water and oil-water interfaces because of their ability to stabilize foams and emulsions. Mixed protein/polysaccharide adsorbed layers at air-water interfaces can be prepared either by adsorption of soluble protein/polysaccharide complexes or by sequential adsorption of complexes or polysaccharides to a previously formed protein layer. Even though the final protein and polysaccharide bulk concentrations are the same, the behavior of the adsorbed layers can be very different, depending on the method of preparation. The surface shear modulus of a sequentially formed beta-lactoglobulin/pectin layer can be up to a factor of 6 higher than that of a layer made by simultaneous adsorption. Furthermore, the surface dilatational modulus and surface shear modulus strongly (up to factors of 2 and 7, respectively) depend on the bulk -lactoglobulin/pectin mixing ratio. On the basis of the surface rheological behavior, a mechanistic understanding of how the structure of the adsorbed layers depends on the protein/polysaccharide interaction in bulk solution, mixing ratio, ionic strength, and order of adsorption to the interface (simultaneous or sequential) is derived. Insight into the effect of protein/polysaccharide interactions on the properties of adsorbed layers provides a solid basis to modulate surface rheological behavior.     

Surface rheology of PEO-PPO-PEO triblock copolymers at the air-water interface: comparison of spread and adsorbed layers

The dilatational rheological properties of monolayers of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide)-type block copolymers at the air-water interface have been investigated by employing an oscillating ring trough method. The properties of adsorbed monolayers were compared to spread layers over a range of surface concentrations. The studied polymers were PEO26-PPO39-PEO26 (P85), PEO103-PPO40-PEO103 (F88), and PEO99-PPO65-PEO99 (F127). Thus, two of the polymers have similar PPO block size and two of them have similar PEO block size, which allows us to draw conclusions about the relationship between molecular structure and surface dilatational rheology. The dilatational properties of adsorbed monolayers were investigated as a function of time and bulk solution concentration. The time dependence was found to be rather complex, reflecting structural changes in the layer. When the dilatational modulus measured at different concentrations was replotted as a function of surface pressure, one unique master curve was obtained for each polymer. It was found that the dilatational behavior of spread (Langmuir) and adsorbed (Gibbs) monolayers of the same polymer is close to identical up to surface concentrations of approximately 0.7 mg/m2. At higher coverage, the properties are qualitatively alike with respect to dilatational modulus, although some differences are noticeable. Relaxation processes take place mainly within the interfacial layers by a redistribution of polymer segments. Several conformational transitions were shown to occur as the area per molecule decreased. PEO desorbs significantly from the interface at segmental areas below 20 A(2), while at higher surface coverage, we propose that segments of PPO are forced to leave the interface to form a mixed sublayer in the aqueous region.     

Interfacial rheological studies of gelatin-sodium dodecyl sulfate complexes adsorbed at the air-water interface

The dilational rheological behavior of gelatin molecules adsorbed at the air-water interface has been studied as a function of sodium dodecyl sulfate (SDS) concentration for a 7 wt % gelatin-SDS solution at 40 degrees C. Binding of SDS molecules to the gelatin strands disrupts the cross-linked network structure of adsorbed gelatin molecules and results in a reduction of the surface elastic modulus of the adsorbed layer that continues until the bulk SDS concentration reaches 1 mM. Beyond this SDS concentration, the dilational rheological properties of the adsorbed gelatin layer are indistinguishable from those of pure SDS adsorbed layers.     

Protein-lipid interactions at the oil-water interface

The distribution of proteins and lipids in food emulsions and foams is determined by competitive and cooperative adsorption between the two types of emulsifiers at the fluid-fluid interfaces, and by the nature of protein-lipid interactions, both at the interface and in the bulk phase. The existence of protein-lipid interactions can have a pronounced impact on the surface rheological properties of these systems. Therefore, these results are of practical importance for food emulsion formulation, texture, and stability. In this study, the existence of protein-lipid interactions at the interface was determined by surface dynamic properties (interfacial tension and surface dilational modulus). Systematic experimental data on surface dynamic properties, as a function of time and at long-term adsorption, for protein (whey protein isolate (WPI)), lipids (monoglycerides), and protein-lipid mixed films at the oil-water interface were measured in an automated drop tensiometer. The dynamic behaviour of protein+lipid mixed films depends on the adsorption time, the lipid and the protein/lipid ratio in a rather complicated manner. The protein determined the interfacial characteristics of the mixed film as the protein at WPI>/=10(-2)% wt/wt saturated the film, no matter what the concentration of the lipid. However, there exists a competitive or cooperative adsorption of the emulsifier (WPI and monoglycerides), as the concentration of protein in the bulk phase is far lower than that for interfacial saturation.     

Disruption of viscoelastic beta-lactoglobulin surface layers at the air-water interface by nonionic polymeric surfactants

Nonequilibrium interfacial layers formed by competitive adsorption of beta-lactoglobulin and the nonionic triblock copolymer PEO99-PPO65-PEO99 (F127) to the air-water interface were investigated in order to explain the influence of polymeric surfactants on protein film surface rheology and foam stability. Surface dilatational and shear rheological methods, surface tension measurements, dynamic thin-film measurements, diffusion measurements (from fluorescence recovery after photo bleaching), and determinations of foam stability were used as methods. The high surface viscoelasticity, both the shear and dilatational, of the protein films was significantly reduced by coadsorption of polymeric surfactant. The drainage rate of single thin films, in the presence of beta-lactoglobulin, increased with the amount of added F127, but equilibrium F127 films were found to be thicker than beta-lactoglobulin films, even at low concentration of the polymeric surfactant. It is concluded that the effect of the nonionic triblock copolymer on the interfacial rheology of beta-lactoglobulin layers is similar to that of low molecular weight surfactants. They differ however in that F127 increases the thickness of thin liquid films. In addition, the significant destabilizing effect of low molecular weight surfactants on protein foams is not found in the investigated system. This is explained as due to long-range steric forces starting to stabilize the foam films at low concentrations of F127.     

Surface adsorption behaviour of milk whey protein and pectin mixtures under conditions of air-water interface saturation

Milk whey proteins (MWP) and pectins (Ps) are biopolymer ingredients commonly used in the manufacture of colloidal food products. Therefore, knowledge of the interfacial characteristics of these biopolymers and their mixtures is very important for the design of food dispersion formulations (foams and/or emulsions). In this paper, we examine the adsorption and surface dilatational behaviour of MWP/Ps systems under conditions in which biopolymers can saturate the air-water interface on their own. Experiments were performed at constant temperature (20 °C), pH 7 and ionic strength 0.05 M. Two MWP samples, β-lactoglobulin (β-LG) and whey protein concentrate (WPC), and two Ps samples, low-methoxyl pectin (LMP) and high-methoxyl pectin (HMP) were evaluated. The contribution of biopolymers (MWP and Ps) to the interfacial properties of mixed systems was evaluated on the basis of their individual surface molecular characteristics. Biopolymer bulk concentration capable of saturating the air-water interface was estimated from surface pressure isotherms. Under conditions of interfacial saturation, dynamic adsorption behaviour (surface pressure and dilatational rheological characteristics) of MWP/Ps systems was discussed from a kinetic point of view, in terms of molecular diffusion, penetration and configurational rearrangement at the air-water interface. The main adsorption mechanism in MWP/LMP mixtures might be the MWP interfacial segregation due to the thermodynamic incompatibility between MWP and LMP (synergistic mechanism); while the interfacial adsorption in MWP/HMP mixtures could be characterized by a competitive mechanism between MWP and HMP at the air-water interface (antagonistic mechanism). The magnitude of these phenomena could be closely related to differences in molecular composition and/or aggregation state of MWP (β-LG and WPC).     

Mechanical properties of interfacial films formed by lysozyme self-assembly at the air-water interface

We present the first characterization of the mechanical properties of lysozyme films formed by self-assembly at the air-water interface using the Cambridge interfacial tensiometer (CIT), an apparatus capable of subjecting protein films to a much higher level of extensional strain than traditional dilatational techniques. CIT analysis, which is insensitive to surface pressure, provides a direct measure of the extensional stress-strain behavior of an interfacial film without the need to assume a mechanical model (e.g., viscoelastic), and without requiring difficult-to-test assumptions regarding low-strain material linearity. This testing method has revealed that the bulk solution pH from which assembly of an interfacial lysozyme film occurs influences the mechanical properties of the film more significantly than is suggested by the observed differences in elastic moduli or surface pressure. We have also identified a previously undescribed pH dependency in the effect of solution ionic strength on the mechanical strength of the lysozyme films formed at the air-water interface. Increasing solution ionic strength was found to increase lysozyme film strength when assembly occurred at pH 7, but it caused a decrease in film strength at pH 11, close to the pI of lysozyme. This result is discussed in terms of the significant contribution made to protein film strength by both electrostatic interactions and the hydrophobic effect. Washout experiments to remove protein from the bulk phase have shown that a small percentage of the interfacially adsorbed lysozyme molecules are reversibly adsorbed. Finally, the washout tests have probed the role played by additional adsorption to the fresh interface formed by the application of a large strain to the lysozyme film and have suggested the movement of reversibly bound lysozyme molecules from a subinterfacial layer to the interface.     

Foams and surface rheological properties of β-casein, gliadin and glycinin

Interfacial rheological properties and their suitability for foam production and stability of two vegetable proteins were studied and compared to β-casein. Proteins used ranged from flexible to rigid/globular in the order of β-casein, gliadin and soy glycinin. Experiments were performed at pH 6.7. Network forming properties were characterised by the surface dilational modulus (determined with the ring trough) and the critical falling film length (L_still) at which a stagnant protein film will break. Gliadin had the highest dilational modulus, followed by glycinin and β-casein, whereas glycinin formed the strongest film against fracture in the overflowing cylinder. The rate of decrease in the surface tension was studied at the air–water (Wilhelmy plate method) and the oil–water interface (bursting membrane) and the dynamic surface tension during compression and expansion in the caterpillar. Gliadin had the lowest equilibrium interfacial tensions and β-casein the lowest dynamic surface tension during expansion. Hardly any foam could be formed at a concentration of 0.1 g/l by shaking. At a concentration of 1.4 g/l most foam was formed by β-casein, followed by gliadin and glycinin. It seems that in the first place the rate of adsorption is important for foam formation. For the vegetable proteins, adsorption was slow. This resulted in lower foamability, especially for glycinin.     

Morphological changes in adsorbed protein films at the oil-water interface subjected to compression, expansion, and heat processing

Adsorbed films of milk proteins at the oil-water (O-W) interface have been imaged using a Brewster angle microscope (BAM). Special adaptations were made to the BAM to allow imaging of the O-W interface and to enable in situ heating and cooling of the adsorbed films. The proteins beta-lactoglobulin (beta-L) and alphas1-, beta-, and kappa-casein were studied over a range of bulk protein concentrations (Cb) and surface ages at pH 7 and for beta-L at pH 5 also. The adsorbed films were subjected to incremental compression and expansion cycles, such that the film area was typically varied between 125% and 50% of the original film area, and the resulting film structure was recorded via the BAM at 25.0 degrees C. Structuring of beta-L films (the formation of ridges and cracks) was more pronounced at pH 5 (closer to the protein's isoelectric point) than at pH 7 and for longer adsorption times and/or higher Cb. Structuring was also much more apparent at the O-W interface than at the A-W interface on compression/expansion/aging, especially at pH 7. After heating beta-L films adsorbed at low Cb (0.005 wt %) to 80 or 90 degrees C, an even greater degree of film structuring was evident, but beta-L films adsorbed at higher Cb (> or =0.05 wt %) showed fewer but larger fractures. The adsorbed caseins showed little evidence of such features, either before or after heating, apart from slight structuring for the heated films of alphas1- and kappa-casein films after 1 day. Changes in the dilatational elastic modulus of the beta-L films (Cb = 0.005 wt %) were correlated with the variations in the structural integrity of the films as observed via the BAM technique. In particular, there was a marked increase in the elastic modulus on heating, while the cycle of compression and expansion appeared to result in a net film weakening overall. The beta-L films adsorbed at higher Cb (> or =0.05 wt %) behaved as if an even stronger elastic skin completely covered the interface. The overall conclusion is that interfacial protein films subjected to these types of thermal and mechanical perturbations, which are typical of those that occur in food colloid processing, can become highly inhomogeneous, depending on the type of protein and the bulk solution conditions. This undoubtedly has implications for the stability of the corresponding emulsions and foams.     

Surface dilatational moduli of poly(vinyl acetate) (PVAc) and PVAc-poly(n-hexyl isocyanate) (PHIC) blend films at the air-water interface

Surface dilatational moduli of poly(vinyl acetate) (PVAc) film and blend films of PVAc and poly(n-hexyl isocyanate) (PHIC) were measured at the air-water interface. PVAc formed a film that was looser and also more stable against strain than the PHIC film. The apparent surface dilatational modulus and surface pressure of the blend films were superimposed on the lower concentration of PVAc, irrespective of the composition of PVAc. However, the additivity rule was not applicable to the apparent surface dilatational modulus and surface pressure. The scaling exponents of the apparent surface dilatational modulus against the added surface concentration decreased with an increase in the proportion of PVAc, suggesting that blend films gradually change from glass material to expanded films.     

Dilatational rheology of beta-casein adsorbed layers at liquid-fluid interfaces

The rheological behavior of beta-casein adsorption layers formed at the air-water and tetradecane-water interfaces is studied in detail by means of pendant drop tensiometry. First, its adsorption behavior is briefly summarized at both interfaces, experimentally and also theoretically. Subsequently, the experimental dilatational results obtained for a wide range of frequencies are presented for both interfaces. An interesting dependence with the oscillation frequency is observed via the comparative analysis of the interfacial elasticity (storage part) and the interfacial viscosity (loss part) for the two interfaces. The analysis of the interfacial elasticities provides information on the conformational transitions undergone by the protein upon adsorption at both interfaces. The air-water interface shows a complex behavior in which two maxima merge into one as the frequency increases, whereas only a single maximum is found at the tetradecane interface within the range of frequencies studied. This is interpreted in terms of a decisive interaction between the oil and the protein molecules. Furthermore, the analysis of the interfacial viscosities provides information on the relaxation processes occurring at both interfaces. Similarly, substantial differences arise between the gaseous and liquid interfaces and various possible relaxation mechanisms are discussed. Finally, the experimental elasticities obtained for frequencies higher than 0.1 Hz are further analyzed on the basis of a thermodynamic model. Accordingly, the nature of the conformational transition given by the maximum at these frequencies is discussed in terms of different theoretical considerations. The formation of a protein bilayer at the interface or the limited compressibility of the protein in the adsorbed state are regarded as possible explanations of the maximum.     

Morphological changes in adsorbed protein films at the air-water interface subjected to large area variations, as observed by brewster angle microscopy

Adsorbed films of proteins at the air-water interface have been imaged using Brewster angle microscopy (BAM). The proteins beta-lactoglobulin (beta-L) and ovalbumin (OA) were studied at a range of protein concentrations and surface ages at 25.0 degrees C and two pH values (7 and 5) in a Langmuir trough. The adsorbed films were periodically subjected to compression and expansion cycles such that the film area was typically varied between 125% and 50% of the original film area. With beta-L on its own, no structural changes were observable at pH 7. When a low-area fraction (less than 0.01%) of 20 mum polystyrene latex particles was spread at the interface before adsorption of beta-L, the particles became randomly distributed throughout the interface, but after protein adsorption and compression/expansion, the particles highlighted notable structural features not visible in their absence. Such features included the appearance of long (several hundred micrometers or more) folds and cracks in the films, generally oriented at right angles to the direction of compression, and also aggregates of protein and/or particles. Such structuring was more visible the longer the film was aged or at higher initial protein concentrations for shorter adsorption times. At pH 5, close to the isoelectric pH of beta-L, such features were just noticeable in the absence of particles but were much more pronounced than at pH 7 in the presence of particles. Similar experiments with OA revealed even more pronounced structural features, both in the absence and presence of particles, particularly at pH 5 (close to the isoelectric pH of OA also), producing striking stripelike and meshlike domains. Changes in the dilatational elasticity of the films could be correlated with the variations in the structural integrity of the films as observed via BAM. The results indicate that interfacial area changes of this type, typical of those that occur in food colloid processing, will lead to highly inhomogeneous adsorbed protein layers, with implications for the stability of the corresponding foams and emulsions stabilized by such films. Overall, the experimental results are in broad agreement with the sorts of trends predicted by earlier computer simulations of protein films subjected to such compression and expansion.     

Interactions in the aqueous phase and adsorption at the air–water interface of caseinoglycomacropeptide (GMP) and β-lactoglobulin mixed systems

The aim of this work was to study the interactions and adsorption of caseinoglycomacropeptide (GMP) and GMP:β-lactoglobulin (β-lg) mixed system in the aqueous phase and at the air–water interface. The existence of associative interactions between GMP and β-lg in the aqueous phase was investigated by dynamic light scattering, differential scanning calorimetry (DSC), fluorometry and native PAGE-electrophoresis. The surface pressure isotherm and the static and dynamic surface pressure were determined by tensiometry and surface dilatational properties. The results showed that GMP presented higher surface activity than β-lg at a concentration of 4% wt but β-lg showed higher film forming ability. In the mixed systems β-lg dominated the static and dynamic surface pressure and the rheological properties of interfacial films suggesting that β-lg hinders GMP adsorption because, in simple competition, GMP should dominate because of its higher surface activity. The surface predominance of β-lg can be attributed to binding of GMP to β-lg in the aqueous phase that prevents GMP adsorption on its own.     

Interfacial and foaming properties of prolylenglycol alginates. Effect of degree of esterification and molecular weight

In the present work we have studied the characteristics of propylene glycol alginates (PGA) adsorption at the air–water interface and the viscoelastic properties of the films in relation to its foaming properties. To evaluate the effect of the degree of PGA esterification and viscosity, different commercial samples were studied—Kelcoloid O (KO), Kelcoloid LVF (KLVF) and Manucol ester (MAN). The temperature (20 °C) and pH (7.0) were maintained constant. For time-dependent surface pressure measurements and surface dilatational properties of adsorbed PGA at the air–water interface an automatic drop tensiometer was used. The foam was generated by whipping and then the foam capacity and stability was determined. The results reveal a significant interfacial activity for PGA due to the hydrophobic character of the propylene glycol groups. The kinetics of adsorption at the air–water interface can be monitored by the diffusion and penetration of PGA at the interface. The adsorbed PGA film showed a high viscoelasticity. The surface dilatational modulus depends on the PGA and its concentration in the aqueous phase. Foam capacity of PGA solutions increased in the order KO > MAN > KLVF, which followed the increase in surface pressure and the decrease in the viscosities of PGA solutions. The stability of PGA foams monitored by the drainage rate and collapse time follows the order MAN > KLVF > KO. The foam stability depends on the combined effect of molecular weight/degree of esterification of PGA, solution viscosity and viscoelasticity of the adsorbed PGA film.     

Dilational viscoelasticity of PEO-PPO-PEO triblock copolymer films at the air-water interface in the range of high surface pressures

The dynamic dilational elasticity of adsorbed and spread films of PEO-PPO-PEO triblock copolymers at the air-water interface was measured as a function of surface pressure, surface age, and frequency. At low surface pressures (<10 mN/m), the surface viscoelasticity is identical to that of PEO homopolymer films. The results at higher surface pressures can be explained by the desorption of PPO segments from the interface and then mixing with PEO segments in water. Unlike some recent results, the spread and adsorbed films are not identical. Spread films exhibit a maximum real part of the dynamic surface elasticity of about 20 mN/m and probably begin to dissolve in water at surface pressures above 19 mN/m. However, the surface elasticity of the adsorbed films decreases beyond the maximum, indicating the formation of a loose surface structure.     

The effect of temperature on food emulsifiers at fluid-fluid interfaces

Heat-induced interfacial aggregation of a whey protein isolate (WPI) with a high content of beta-lactoglobulin (>92%), previously adsorbed at the oil-water interface, was studied by means of interfacial dynamic characteristics performed in an automatic drop tensiometer. Protein concentration in aqueous bulk phase ranging between 1x10(-1) and 1x10(-5) % wt/wt was studied as a variable. The experiments were carried out at temperatures ranging from 20-80 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. Interfacial crystallisation of food polar lipids (monopalmitin, monoolein, and monolaurin) previously adsorbed at the oil-water interface, was studied by interfacial dynamic characteristics (interfacial tension and surface dilational properties). The temperature, ranging between 40 and 2 degrees C, and the lipid concentration in aqueous oil phase, ranging between 1x10(-2) and 1x10(-4) % wt/wt, were studied as variables. Significant changes in interfacial dynamic characteristics associated with interfacial lipid crystallisation were observed as a function of lipid concentration in the bulk phase. Interfacial crystallisation of food polar lipids (monopalmitin, monoolein, and monolaurin) at the air-water interface, was studied by pi-A isotherms performed in a Langmuir trough coupled with Brewster angle microscopy (BAM). A condensation in monoglyceride monolayers towards lower molecular area was observed as the temperature decreased. This effect was attributed to lipid crystallisation at lower temperatures. BAM images corroborated the effect of temperature on the monolayer structure, as a function of the monoglyceride type.     

Cross linking and rheological characterization of adsorbed protein layers at the oil-water interface

The dilatational rheological properties of cross-linked protein layers adsorbed at the oil-water interface were investigated with help of a modified drop tensiometer allowing successive replacements of the external phase. This setup enables one to perform cross-linking reactions at the interface only, that is, without any contact between the cross-linking agent and protein molecules in solution, under continuous monitoring of the interfacial tension. The mechanical properties of the resulting interface were investigated with dilatational large strain experiments. Measured rheological properties were related to the expected stability of an emulsion against disproportionation by considering the ratio of the interfacial elasticity to the interfacial tension. In an attempt to increase this ratio to improve the resistance against disproportionation, experiments were performed with densified protein layers obtained via reduction of the droplet area prior to cross linking. To highlight the influence of the protein morphology on the dilatational rheological properties of the cross-linked adsorbed layers, experiments were performed with random coil (beta-casein) as well as globular (beta-lactoglobulin) proteins. Glutaraldehyde was used as a cross-linking agent. Experiments were performed at 55 degrees C and pH 7.0 in 20 mM imidazole buffer for later comparison with enzymatically cross-linked adsorbed protein layers. The present work demonstrated substantial qualitative and quantitative differences in the interfacial rheological properties of cross-linked random coil and globular proteins.     

Polysaccharide charge density regulating protein adsorption to air/water interfaces by protein/polysaccharide complex formation

Because the formation of protein/polysaccharide complexes is dominated by electrostatic interaction, polysaccharide charge density is expected to play a major role in the adsorption behavior of the complexes. In this study, pullulan (a non-charged polysaccharide) carboxylated to four different charge densities (fraction of carboxylated subunits: 0.1, 0.26, 0.51, and 0.56) was used to investigate the effect of charge density on the properties of mixed protein/polysaccharide adsorbed layers at air/water interfaces. With all pullulan samples, soluble complexes with beta-lactoglobulin could be formed at low ionic strength, pH 4.5. It was shown that the higher was the pullulan charge density, the more the increase of surface pressure in time was retarded as compared to that for pure beta-lactoglobulin. The retardation was even more pronounced for the development of the dilatational modulus. The lower dilatational modulus can be explained by the ability of the polysaccharides to prevent the formation of a compact protein layer at the air/water interface due to electrostatic repulsion. This ability of the polysaccharides to prevent "layer compactness" increases with the net negative charge of the complexes. If charge density is sufficient (> or = 0.26), polysaccharides may enhance the cohesion between complexes within the adsorbed layer. The charge density of polysaccharides is shown to be a dominant regulator of both the adsorption kinetics as well as the resulting surface rheological behavior of the mixed layers formed. These findings have significant value for the application of complex protein-polysaccharide systems.