Shear and dilatational relaxation mechanisms of globular and flexible proteins at the hexadecane/water interface

Proteins adsorbed at fluid/fluid interfaces influence many phenomena: food emulsion and foam stability (Murray et al. Langmuir 2002, 18, 9476 and Borbas et al. Colloids Surf., A 2003, 213, 93), two-phase enzyme catalysis (Cascao-Pereira et al. Biotechnol. Bioeng. 2003, 83, 498; 2002, 78, 595), human lung function (Lunkenheimer et al. Colloids Surf., A 1996, 114, 199; Wustneck et al.; and Banerjee et al. 2000, 15, 14), and cell membrane mechanical properties (Mohandas et al. 1994, 23, 787). Time scales important to these phenomena are broad, necessitating an understanding of the dynamics of biological macromolecules at interfaces. We utilize interfacial shear and dilatational deformations to study the rheology of a globular protein, lysozyme, and a disordered protein, beta-casein, at the hexadecane/water interface. Linear viscoelastic properties are measured using small amplitude oscillatory flow, stress relaxation after a sudden dilatational displacement, and shear creep response to probe the rheological response over broad experimental time scales. Our studies of lysozyme and beta-casein reveal that the interfacial dissipation mechanisms are strongly coupled to changes in the protein structure upon and after adsorption. For beta-casein, the interfacial response is fluidlike in shear deformation and is dominated by interfacial viscous dissipation, particularly at low frequencies. Conversely, the dilatational response of beta-casein is dominated by diffusion dissipation at low frequencies and viscous dissipation at higher frequencies (i.e., when the experimental time scale is faster than the characteristic time for diffusion). For lysozyme in shear deformation, the adsorbed protein layer is primarily elastic with only a weak frequency dependence. Similarly, the interfacial dilatational moduli change very little with frequency. In comparison to beta-casein, the frequency response of lysozyme does not change substantially after washing the protein from the bulk solution. Apparently, it is the irreversibly adsorbed fraction that dominates the dynamic rheological response for lysozyme. Using stress relaxation after a sudden dilatational displacement and shear creep response, the characteristic time of relaxation was found to be 1000 s in both modes of deformation. The very long relaxation time for lysozyme likely results from the formation of a glassy interfacial network. This network develops at high interfacial concentrations where the molecules are highly constrained because of conformation changes that prevent desorption.     

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.     

Effect of thermal treatment on interfacial properties of beta-lactoglobulin

The changes in the secondary conformation and surface hydrophobicity of beta-lactoglobulin subjected to different thermal treatments were characterized at pH values of 7, 5.5 and 4 using circular dichroism (CD) and hydrophobic dye binding. Heating resulted in a decrease in alpha-helix content with a corresponding increase in random coil at all pH values, this change being more pronounced for small heating times. Heating also resulted in an increase in surface hydrophobicity as a result of partial denaturation, this increase being more pronounced at pH 4. Thermal treatment resulted in a shift of the spread monolayer isotherm at air-water interface to smaller area per molecule due to increased flexibility and more loop formation. Thermal treatment led to an increase in interfacial shear elasticity and viscosity of adsorbed beta-lactoglobulin layer at pH 5.5 and 7. Interfacial shear elasticity, shear viscosity, stability of beta-lactoglobulin stabilized emulsion and average coalescence time of a single droplet at a planar oil-water interface with adsorbed protein layer exhibited a maximum for protein subjected to 15 min heat treatment at pH 7. At pH 5.5, the interfacial shear rheological properties and average single drop coalescence time were maximum for 15 min heat treatment whereas emulsion stability was maximum for 5 min heat treatment. At pH 7, thermal treatment was found to enhance foam stability. Analysis of thin film drainage indicated that interfacial shear rheological properties do not influence thin film drainage.     

Stability of thin stagnant film on a solid surface with a viscoelastic air-liquid interface

Linear stability analysis for a film on a solid surface with a viscoelastic air-liquid interface is presented. The interfacial dilatational and shear viscoelastic properties were described by Maxwell models. Dilatational and shear interfacial elasticity and viscosity were shown to improve film stability. When the interfacial rheological properties are extremely large or small, the maximum perturbation growth coefficient is shown to reduce to those for immobile and mobile interfaces respectively. Calculated values of maximum growth coefficient for thin film stabilized by 0.5% beta-lactoglobulin approached those of mobile films for thick (>2000 nm) and those for immobile films for thin (<100 nm) films respectively with the values lying between the two limits for intermediate film thicknesses.     

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.     

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.     

Interfacial behaviour and mechanical properties of spread lung surfactant protein/lipid layers

The surface behaviour of spread dipalmitoyl phosphatidyl choline (DPPC), lung surfactant protein C (SP-C), and their mixtures were characterised using a captive bubble surfactometer. The surface tension was determined by using axisymmetric bubble shape analysis. Surface dilatational rheological behaviour was characterised by sinusoidal oscillation of the bubble volume and at frequencies 0.006-0.025 Hz. The pi/A isotherms of DPPC, SP-C, and their mixtures were described with a generalised equation of state. Monolayer cycling of mixed DPPC/SP-C layers yields isotherms with a plateau in the range of 50-53 mN/m. When the surface pressure becomes higher SP-C is squeezed out of the film, but it re-enters the film upon expansion. Surface dilatational elasticities of DPPC films had a maximum at about 30 mN/m. At higher surface pressures, the films became brittle and the elasticity decreased. A slightly pronounced maximum was found at a surface pressure exceeding 55 mN/m. The dilatational viscosity had two distinct maxima, corresponding with those in the elasticity curves, i.e. one before the minimum area demand, and one in the range of over-compression. This was explained by the formation of a second ordered complex structure in the range of film over-compression. SP-C films show continuously increasing dilatational elasticities and viscosities with a maximum at f approximately 0.02 Hz. Mixed monolayers, DPPC+2 mol% SP-C, had dilatational elasticities increasing with surface pressure. In contrast to DPPC alone, an elasticity maximum appeared in the range of the squeeze out plateau. The dilatational viscosity had two distinct maxima as observed for DPPC, whereas the maximum before the squeeze out plateau is very broad like that of SP-C. The viscosity decreased for frequencies higher 0.02 Hz favouring elastic properties of the film. Our data provide experimental evidence that SP-C mixed with DPPC yield higher elasticities and viscosities as compared with films formed by the single components. This behaviour is likely to support breathing cycles, especially for the turn from inspiration to expiration and vice versa.     

Interfacial layers from the protein HFBII hydrophobin: dynamic surface tension, dilatational elasticity and relaxation times

The pendant-drop method (with drop-shape analysis) and Langmuir trough are applied to investigate the characteristic relaxation times and elasticity of interfacial layers from the protein HFBII hydrophobin. Such layers undergo a transition from fluid to elastic solid films. The transition is detected as an increase in the error of the fit of the pendant-drop profile by means of the Laplace equation of capillarity. The relaxation of surface tension after interfacial expansion follows an exponential-decay law, which indicates adsorption kinetics under barrier control. The experimental data for the relaxation time suggest that the adsorption rate is determined by the balance of two opposing factors: (i) the barrier to detachment of protein molecules from bulk aggregates and (ii) the attraction of the detached molecules by the adsorption layer due to the hydrophobic surface force. The hydrophobic attraction can explain why a greater surface coverage leads to a faster adsorption. The relaxation of surface tension after interfacial compression follows a different, square-root law. Such behavior can be attributed to surface diffusion of adsorbed protein molecules that are condensing at the periphery of interfacial protein aggregates. The surface dilatational elasticity, E, is determined in experiments on quick expansion or compression of the interfacial protein layers. At lower surface pressures (<11 mN/m) the experiments on expansion, compression and oscillations give close values of E that are increasing with the rise of surface pressure. At higher surface pressures, E exhibits the opposite tendency and the data are scattered. The latter behavior can be explained with a two-dimensional condensation of adsorbed protein molecules at the higher surface pressures. The results could be important for the understanding and control of dynamic processes in foams and emulsions stabilized by hydrophobins, as well as for the modification of solid surfaces by adsorption of such proteins.     

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.     

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.     

Dynamic properties of soy globulin adsorbed films at the air-water interface

In this paper we present surface dilatational properties of soy globulins (beta-conglycinin, glycinin, and reduced glycinin with 10 mM of dithiothreitol (DTT)) adsorbed onto the air-water interface, as a function of adsorption time. The experiments were performed at constant temperature (20 degrees C), pH (8.0), and ionic strength (0.05 M). The surface rheological parameters were measured as a function of protein concentration (ranging from 1 to 1x10(-3)% wt/wt). We found that the surface dilatational modulus, E, increases, and the phase angle, phi, decreases with time, theta, which may be associated with protein adsorption. These phenomena have been related to protein adsorption, unfolding, and/or protein-protein interactions (at long-term adsorption) as a function of protein concentration in solution. From a rheological point of view, the surface viscoelastic characteristics of soy globulin films adsorbed at the air-water interface are practically elastic. The main conclusion is that the dilatational properties of the adsorbed films depend on the molecular structure of the protein.     

Electrostatic effects on the yield stress of whey protein isolate foams

The mechanisms responsible for foam structure are of practical interest within the food industry. The yield stress (tau) of whey protein isolate (WPI) foams as affected by electrostatic forces was investigated by whipping 10% (w/v) protein solutions prepared over a range of pH levels and salt concentrations. Measurements of foam overrun and model WPI interfaces, i.e. adsorption kinetics as determined via dynamic surface tension and dilatational rheological characterization, aided data interpretation. Interfacial measurements were also made with the primary whey proteins, beta-lactoglobulin (beta-lg) and alpha-lactalbumin (alpha-la). Yield stress of WPI foams was dependent on pH, salt type and salt concentration. In the absence of salt, tau was highest at pH 5.0 and lowest at pH 3.0. The addition of NaCl and CaCl2 up to 400 mM significantly increased tau at pH 7.0 but not at pH 3.0. Furthermore, at pH 7.0, equivalent molar concentrations of CaCl2 as compared to NaCl increased tau to greater extents. Salts had minimal effects on tau at pH 5.0. Comparisons with interfacial rheological data suggested the protein's capacity to contribute towards tau was related to the protein's potential at forming strong, elastic interfaces throughout the structure. The dynamic surface tension data for beta-lg and alpha-la were similar to WPI, while the interfacial rheological data displayed several noticeable differences.     

Dilatational rheology and foaming properties of sucrose monoesters in the presence of beta-lactoglobulin

The foaming properties and the dilatational rheology of systems containing purified sucrose caprylate (SM800), caprate (SM1000), laurate (SM1200) and palmitate (SM1600) have been studied. Addition of beta-lactoglobulin (beta-lg) at a low concentration (0.050 wt.%) can aid the foam formation in the cases, where the surfactant concentration is insufficient to support foam formation. However, foams where both species co-existed exhibited poor stability. beta-lg was found to affect the dilatational properties of surfactant films even at low concentrations. It is thought that this could be related to the effect of the protein on the adsorption-desorption relaxation mechanism, or to the possible formation of a protein-surfactant complex in the bulk. The age of the protein film was also found to affect the kinetics of protein displacement by SM1000, as monitored by the change in the dilatational properties (Langmuir trough technique) and the relative reflectivity of the interface (Brewster angle microscopy) with time. An insoluble monolayer of sucrose stearate (Suc18) and beta-lg was also studied and it was found that the presence of small amounts of Suc18 in the protein film lead to a reduction of the interfacial elasticity. This is believed to be due to the disruption of the protein network. A possible mechanism could involve the obstruction of the hydrogen-bond intermolecular protein association by the strongly hydrated sucrose headgroup or the obstruction of the protein-protein hydrophobic interactions by the formation of an interfacial protein-surfactant complex.     

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   

Interfacial Behavior of beta-Lactoglobulin at a Stainless Steel Surface: An Electrochemical Impedance Spectroscopy Study

The electrochemical impedance spectroscopy technique was used to investigate the interfacial behavior of beta-lactoglobulin at an austenitic stainless steel surface over the temperature range 299 to 343 K at an open circuit potential. The electrode/electrolyte interface and corresponding surface processes were successfully modeled by applying an equivalent-electrical-circuit approach. A charge-transfer resistance value was found to be very sensitive to the amount of adsorbed protein (surface concentration), thus indicating that the adsorption of the protein (i) was accompanied by the transfer of the charge, via chemisorption, and (ii) influenced the mechanism and kinetics of the corrosion reaction. This was also apparent from the large decrease in the corrosion activation energy (16 kJ mol(-1)) caused by the adsorption of the protein. Adsorption of beta-lactoglobulin onto the stainless steel surface at an open circuit potential resulted in a unimodal isotherm at all the temperatures studied and the adsorption process was described with a Langmuir adsorption isotherm. From the calculated Gibbs free energies of adsorption it was confirmed that beta-lactoglobulin molecules adsorb strongly onto the stainless steel surface. The enthalpy and entropy values indicated that the molecule partially unfolds at the surface upon adsorption. The adsorption process was found to be entirely governed by the change in entropy. Copyright 2000 Academic Press.     

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.     

短链烷基对多取代烷基苯磺酸盐界面性质的影响

利用悬挂滴方法研究了2,5-二乙基-4-壬基苯磺酸钠(292)、2,5-二丙基-4-壬基苯磺酸钠(393)和2,5-二丁基-4-壬基苯磺酸钠(494)在空气-水表面和正癸烷-水界面的扩张流变性质,考察了时间、界面压、工作频率及体相浓度对扩张弹性和粘性的影响。研究发现,在低表面活性剂浓度条件下,表面吸附膜类似弹性膜,其强度由膜内分子的相互作用决定;高浓度下体相与表面间的扩散交换过程控制表面膜的性质。油分子的插入导致界面吸附分子之间相互作用的削弱,扩散交换过程主导界面膜性质;但随着短链烷基长度增加,油分子的影响变小。表面膜的强度在吸附达到平衡前已经决定,而界面膜在吸附饱和后仍然随界面分子重排而变化。     

Interfacial characterization of beta-lactoglobulin networks: displacement by bile salts

The competitive displacement of a model protein (beta-lactoglobulin) by bile salts from air-water and oil-water interfaces is investigated in vitro under model duodenal digestion conditions. The aim is to understand this process so that interfaces can be designed to control lipid digestion thus improving the nutritional impact of foods. Duodenal digestion has been simulated using a simplified biological system and the protein displacement process monitored by interfacial measurements and atomic force microscopy (AFM). First, the properties of beta-lactoglobulin adsorbed layers at the air-water and the olive oil-water interfaces were analyzed by interfacial tension techniques under physiological conditions (pH 7, 0.15 M NaCl, 10 mM CaCl2, 37 degrees C). The protein film had a lower dilatational modulus (hence formed a weaker network) at the olive oil-water interface compared to the air-water interface. Addition of bile salt (BS) severely decreased the dilatational modulus of the adsorbed beta-lactoglobulin film at both the air-water and olive oil-water interfaces. The data suggest that the bile salts penetrate into, weaken, and break up the interfacial beta-lactoglobulin networks. AFM images of the displacement of spread beta-lactoglobulin at the air-water and the olive oil-water interfaces suggest that displacement occurs via an orogenic mechanism and that the bile salts can almost completely displace the intact protein network under duodenal conditions. Although the bile salts are ionic, the ionic strength is sufficiently high to screen the charge allowing surfactant domain nucleation and growth to occur resulting in displacement. The morphology of the protein networks during displacement is different from those found when conventional surfactants were used, suggesting that the molecular structure of the surfactant is important for the displacement process. The studies also suggest that the nature of the oil phase is important in controlling protein unfolding and interaction at the interface. This in turn affects the strength of the protein network and the ability to resist displacement by surfactants.     

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.     

Foaming and interfacial properties of hydrolyzed beta-lactoglobulin

beta-lactoglobulin (beta-lg) was hydrolyzed with three different proteases and subsequently evaluated for its foaming potential. Foam yield stress (tau0) was the primary variable of interest. Two heat treatments designed to inactivate the enzymes, 75 degrees C/30 min and 90 degrees C/15 min, were also investigated for their effects on foam tau0. Adsorption rates and dilatational rheological tests at a model air/water interface aided data interpretation. All unheated hydrolysates improved foam tau0 as compared to unhydrolyzed beta-lg, with those of pepsin and Alcalase 2.4L(R) being superior to trypsin. Heat inactivation negatively impacted foam tau0, although heating at 75 degrees C/30 min better preserved this parameter than heating at 90 degrees C/15 min. All hydrolysates adsorbed more rapidly at the air/water interface than unhydrolyzed beta-lg, as evidenced by their capacity to lower the interfacial tension. A previously observed relationship between interfacial dilatational elasticity (E') and tau0 was generally confirmed for these hydrolysates. Additionally, the three hydrolysates imparting the highest tau0 not only had high values of E' (approximately twice that of unhydrolyzed beta-lg), they also had very low phase angles (essentially zero). This highly elastic interfacial state is presumed to improve foam tau0 indirectly by improving foam stability and directly by imparting resistance to interfacial deformation.