Modulation of the adsorption properties at air-water interfaces of complexes of egg white ovalbumin with pectin by the dielectric constant

The possibility of modulating the mesoscopic properties of food colloidal systems by the dielectric constant is studied by determining the impact of small amounts of ethanol (10%) on the adsorption of egg white ovalbumin onto the air-water interface in the absence and presence of pectin. The adsorption kinetics was monitored using tensiometry. The addition of ethanol resulted in considerably slower adsorption of the protein onto the interface, and this effect was enhanced when the protein was in complex with the pectin. Time-resolved fluorescence measurements demonstrated that in the case of noncomplexed ovalbumin the addition of ethanol resulted in a more condensed protein surface layer where ovalbumin adopted a preferred orientation at the interface. In contrast, the effect of ethanol on the ovalbumin-pectin complex suggested a pronounced multipoint electrostatic interaction between protein and polyelectrolyte and the formation of a more rigid spatial arrangement within the complex, thereby leading to suppressed protein-protein interactions. From this work it is concluded that by the enhanced binding affinity between ovalbumin and pectin a strong effect on the adsorption properties of the protein can be accomplished. This work does therefore illustrate how solvent quality can be exploited effectively to enhance or suppress protein functional behavior in complex applications containing air-water interfaces.     

Structure of mixed beta-lactoglobulin/pectin adsorbed layers at air/water interfaces; a spectroscopy study

Based on earlier reported surface rheological behaviour two factors appeared to be important for the functional behaviour of mixed protein/polysaccharide adsorbed layers at air/water interfaces: (1) protein/polysaccharide mixing ratio and (2) formation history of the layers. In this study complexes of beta-lactoglobulin (positively charged at pH 4.5) and low methoxyl pectin (negatively charged) were formed at two mixing ratios, resulting in negatively charged and nearly neutral complexes. Neutron reflection showed that adsorption of negative complexes leads to more diffuse layers at the air/water interface than adsorption of neutral complexes. Besides (simultaneous) adsorption of protein/polysaccharide complexes, a mixed layer can also be formed by adsorption of (protein/)polysaccharide (complexes) to a pre-formed protein layer (sequential adsorption). Despite similar bulk concentrations, adsorbed layer density profiles of simultaneously and sequentially formed layers were persistently different, as illustrated by neutron reflection analysis. Time resolved fluorescence anisotropy showed that the mobility of protein molecules at an air/water interface is hampered by the presence of pectin. This hampered mobility of protein through a complex layer could account for differences observed in density profiles of simultaneously and sequentially formed layers. These insights substantiated the previously proposed organisations of the different adsorbed layers based on surface rheological data.     

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.     

Effect of H(2)O-CO(2) organization on ovalbumin adsorption at the supercritical CO(2)-water interface

We have studied the formation of water-CO(2) interfaces in the presence of different concentrations of ovalbumin (OVA) by tensiometry and by means of interfacial rheological measurements to obtain some information on the capacity of protein film to stabilize H(2)O in CO(2) emulsion. The formation of pure water-CO(2) interface can be described as a two-step phenomenon.(1) The CO(2) molecules adsorb onto the water surface and then a reorganization of the interface creates a H(2)O-CO(2) cluster network. This organization occurs at a temperature (40 degrees C) higher than the higher temperature limit (10 degrees C) allowing the formation of crystalline structure called CO(2) clathrate.(2) Our results show that ovalbumin adsorption from bulk concentrations higher than 0.0229 g/L inhibits the cluster formation for a CO(2) pressure less than 80 bar. However, for lower concentrations, the more the CO(2) pressure is close to 80 bar, the more OVA adsorption is reduced by the H(2)O-CO(2) cluster network. Moreover, from a pressure of 90 bar, the affinity of OVA for the interface increases and mixed films made of protein molecules and clusters are obtained for the OVA concentrations lower than 1 g/L.     

Interfacial rheology of protein–surfactant mixtures

The distribution of proteins and surfactants at fluid interfaces (air–water and oil–water) is determined by the competitive adsorption between the two types of emulsifiers and by the nature of the protein–surfactant interactions, both at the interface and in the bulk phase, with a pronounced impact on the interfacial rheological properties of these systems. Therefore, the interfacial rheology is of practical importance for food dispersion (emulsion or foam) formulation, texture, and stability. In this review, the existence of protein–surfactant interactions, the mechanical behaviour and/or the composition of emulsifiers at the interface are indirectly determined by interfacial rheology of the mixed films. The effect on the interfacial rheology of protein–surfactant mixed films of the protein, the surfactant, the interface and bulk compositions, the method of formation of the interfacial film, the interactions between film forming components, and the displacement of protein by surfactant have been analysed. The last section tries to understand the role of interfacial rheology of protein–surfactant mixed films on food dispersion formation and stability. The emphasis of the present review is on the interfacial dilatational rheology.     

Dynamic adsorption and characterization of phospholipid and mixed phospholipid/protein layers at liquid/liquid interfaces

Drop profile analysis tensiometry is applied to study the adsorption dynamics of phospholipids, proteins and phospholipid/protein mixtures at liquid/liquid interfaces. Measurements of the dynamic interfacial tension of phospholipid layers give information on the adsorption mechanism and the structure of the adsorption layer. The equilibrium and dynamic adsorption of pure protein solutions, i.e. human serum album (HSA), beta-lactoglobulin (beta-LG), beta-casein (beta-CA), can be explained well by the thermodynamic model of Frumkin and the diffusion-controlled adsorption theory. The adsorption behavior from mixed phospholipid/protein solutions was also investigated in terms of dynamic interfacial tensions. Interestingly, a "skin-like" folded film of pure protein or phospholipid/protein complex layers can be observed at curved surfaces at the water/oil interfaces. The addition of phospholipids accelerates the formation of the folded structure at the drop surface through co-adsorption of proteins.     

Silver nanoplates formed at the air/water and solid/water interfaces

Silver nanoparticles and nanoplates were prepared at the air/AgNO₃ aqueous solution interfaces under poly(9-vinylcarbazole) (PVK) monolayers when illuminated by UV-light at room temperature and elevated temperatures, respectively. When the illuminated films at the air/water interfaces were covered by carbon-coated copper grids, nanoplates were formed even at room temperature, and the size of the nanoplates was much larger than those formed at the air/water interface under the same experimental conditions, indicating that copper took part in the formation of Ag nanoplates through the galvanic displacement reaction between Cu and Ag⁺ ions with the help of carbon layer to conduct electrons. It was found that the basal plane of these nanoplates is the (1 1 1) face of a face-centered cubic (fcc) Ag crystal. Although platelike structure can be formed at the carbon-coated copper grid/AgNO₃ aqueous solution interface without PVK film, it shows different features from those with PVK films, indicating that PVK film plays an important role in the formation of regular large nanoplates. Further observations indicate that special restrained microenvironment, adsorption of PVK molecules on a specific crystal face, anisotropic growth and attachment of the nanoparticles are responsible for the formation of the nanoplates.     

Polysaccharide/Surfactant complexes at the air-water interface - effect of the charge density on interfacial and foaming behaviors

The binding of a cationic surfactant (hexadecyltrimethylammonium bromide, CTAB) to a negatively charged natural polysaccharide (pectin) at air-solution interfaces was investigated on single interfaces and in foams, versus the linear charge densities of the polysaccharide. Besides classical methods to investigate polymer/surfactant systems, we applied, for the first time concerning these systems, the analogy between the small angle neutron scattering by foams and the neutron reflectivity of films to measure in situ film thicknesses of foams. CTAB/pectin foam films are much thicker than the pure surfactant foam film but similar for high- and low-charged pectin/CTAB systems despite the difference in structure of complexes at interfaces. The improvement of the foam properties of CTAB bound to pectin is shown to be directly related to the formation of pectin-CTAB complexes at the air-water interface. However, in opposition to surface activity, there is no specific behavior for the highly charged pectin: foam properties depend mainly upon the bulk charge concentration, while the interfacial behavior is mainly governed by the charge density of pectin. For the highly charged pectin, specific cooperative effects between neighboring charged sites along the chain are thought to be involved in the higher surface activity of pectin/CTAB complexes. A more general behavior can be obtained at lower charge density either by using a low-charged pectin or by neutralizing the highly charged pectin in decreasing pH.     

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.     

Dynamics of mixed protein–surfactant layers adsorbed at the water/air and water/oil interface

Drop and bubble shape tensiometry experiments are performed at the water/air and water/hexane interfaces in order to get more information about the differences in the adsorption layer structure of mixed protein/surfactant systems. For mixtures of β-lactoglobulin and sodium dodecyl sulphate the adsorption at the water/air interface is essentially a competitive process between protein/surfactant complexes and free surfactant molecules, while the water/oil interface is essentially covered by the complexes.     

Role of interfacial water on protein adsorption at cross-linked polyethylene oxide interfaces

Sum frequency generation (SFG) vibrational spectroscopy was used to study the structure of water at cross-linked PEO film interfaces in the presence of human serum albumin (HSA) protein. Although PEO is charge neutral, the PEO film/water interface exhibited an SFG signal of water similar to that of a highly charged water/silica interface, signifying the presence of ordered water. Ordered water molecules were observed not only at the water/PEO interface, but also within the PEO film. It indicates that the PEO and water form an ordered hydrogen-bonded network extending from the bulk PEO film into liquid water, which can provide an energy barrier for protein adsorption. Upon exposure to the protein solution, the SFG spectra of water at the water/PEO interface remained nearly unperturbed. For comparison, the SFG spectra of water/silica and water/polystyrene interfaces were also studied with and without HSA in the solution. The SFG spectra of the interfacial water were correlated with the amount of protein adsorbed on the surfaces using fluorescence microscopy, which showed that the amount of protein adsorbed on the PEO film was about 10 times less than that on a polystyrene film and 3 times less than that on silica.     

Adsorption of naphthalene and ozone on atmospheric air/ice interfaces coated with surfactants: a molecular simulation study

The adsorption of gas-phase naphthalene and ozone molecules onto air/ice interfaces coated with different surfactant species (1-octanol, 1-hexadecanol, or 1-octanal) was investigated using classical molecular dynamics (MD) simulations. Naphthalene and ozone exhibit a strong preference to be adsorbed at the surfactant-coated air/ice interfaces, as opposed to either being dissolved into the bulk of the quasi-liquid layer (QLL) or being incorporated into the ice crystals. The QLL becomes thinner when the air/ice interface is coated with surfactant molecules. The adsorption of both naphthalene and ozone onto surfactant-coated air/ice interfaces is enhanced when compared to bare air/ice interface. Both naphthalene and ozone tend to stay dissolved in the surfactant layer and close to the QLL, rather than adsorbing on top of the surfactant molecules and close to the air region of our systems. Surfactants prefer to orient at a tilted angle with respect to the air/ice interface; the angular distribution and the most preferred angle vary depending on the hydrophilic end group, the length of the hydrophobic tail, and the surfactant concentration at the air/ice interface. Naphthalene prefers to have a flat orientation on the surfactant coated air/ice interface, except at high concentrations of 1-hexadecanol at the air/ice interface; the angular distribution of naphthalene depends on the specific surfactant and its concentration at the air/ice interface. The dynamics of naphthalene molecules at the surfactant-coated air/ice interface slow down as compared to those observed at bare air/ice interfaces. The presence of surfactants does not seem to affect the self-association of naphthalene molecules at the air/ice interface, at least for the specific surfactants and the range of concentrations considered in this study.     

Characterization of the emulsification properties of 2S albumins from sunflower seed

The ability of 2S albumins from sunflower seeds to stabilize oil-in-water emulsions has been investigated, demonstrating that one of the proteins (SFA8) effectively stabilizes emulsions, while another (SF-LTP) does not stabilize emulsions. The surface tension and surface dilation viscosity of these two proteins were measured, rationalizing the emulsifying ability of SFA8 in terms of its ability to form a strongly elastic monolayer at interfaces. The secondary structure changes that occur upon adsorption of SFA8 to the oil/water interface have also been studied by fluorescence, circular dichroism (CD), and Fourier-transform infrared (FT-IR) spectroscopy. It was found that the beta-sheet content of the protein increased upon adsorption at the expense of alpha-helix and random structure. Moreover, FT-IR measurements indicate the presence of intermolecular beta-sheet formation upon adsorption. Fluorescence studies with an oil-soluble fluorescence quencher indicate that the single tryptophan residue present in SFA8 may become located in the oil-phase of the emulsion. This residue is thought to be partially buried in the native protein, and these data suggest that changes in the polypeptide region flanking this residue may play an important role in the molecular rearrangement that occur on or following adsorption to the oil/water interface.     

Polyelectrolyte/surfactant mixtures in the bulk and at water/oil interfaces

Stabilization of emulsions by mixed polyelectrolyte/surfactant systems is a prominent example for the application in modern technologies. The formation of complexes between the polymers and the surfactants depends on the type of surfactant (ionic, non-ionic) and the mixing ratio. The surface activity (hydrophilic–lipophilic balance) of the resulting complexes is an important quantity for its efficiency in stabilizing emulsions. The interfacial adsorption properties observed at liquid/oil interfaces are more or less equivalent to those observed at the aqueous solution/air interface, however, the corresponding interfacial dilational and shear rheology parameters differ quite significantly. The interfacial properties are directly linked to bulk properties, which support the picture for the complex formation of polyelectrolyte/surfactant mixtures, which is the result of electrostatic and hydrophobic interactions. For long alkyl chain surfactants the interfacial behavior is strongly influenced by hydrophobic interactions while the complex formation with short chain surfactants is mainly governed by electrostatic interactions.     

Surface microstructure and cell compatibility of calcium silicate and calcium phosphate composite coatings on Mg-Zn-Mn-Ca alloys for biomedical application

The flexibility and aggregation of proteins can cause adsorption to oil-water interfaces and thereby create challenges during formulation and processing. Protein adsorption is a complex process and the presence of surfactants further complicates the system, in which additional parameters need to be considered. The purpose of this study is to scrutinize the influence of surfactants on protein adsorption to interfaces, using lysozyme as a model protein and sorbitan monooleate 80 (S80), polysorbate 80 (T80), polyethylene-block-poly(ethylene glycol) (PE-PEG) and polyglycerol polyricinoleate (PG-PR) as model surfactants. Rheological properties, measured using a TA AR-G2 rheometer equipped with a double wall ring (DWR) geometry, were used to compare the efficacy of the surfactant in hindering lysozyme adsorption. The system consists of a ring and a Delrin? trough with a circular channel (interfacial area=1882.6 mm(2)). Oscillatory shear measurements were conducted at a constant frequency of 0.1 Hz, a temperature of 25°C, and with strain set to 1%. The adsorption of lysozyme to the oil-water interface results in the formation of a viscoelastic film. This can be prevented by addition of surfactants, in a manner depending on the concentration and the type of surfactant. The more hydrophilic surfactants are more effective in hindering lysozyme adsorption to oil-water interfaces. Additionally, the larger surfactants are more persistent in preventing film formation, whereas the smaller ones eventually give space for the lysozyme on the interface. The addition of a mixture of two different surfactants was only beneficial when the two hydrophilic surfactants were mixed, in which case a delay in the multilayer formation was detected. The method is able to detect the interfacial adsorption of lysozyme and thus the hindering of film formation by model surfactants. It can therefore aid in processing of any delivery systems for proteins in which the protein is introduced to oil-water interfaces.     

Structural rearrangement of β-lactoglobulin at different oil-water interfaces and its effect on emulsion stability

Understanding the factors that control protein structure and stability at the oil-water interface continues to be a major focus to optimize the formulation of protein-stabilized emulsions. In this study, a combination of synchrotron radiation circular dichroism spectroscopy, front-face fluorescence spectroscopy, and dual polarization interferometry (DPI) was used to characterize the conformation and geometric structure of β-lactoglobulin (β-Lg) upon adsorption to two oil-water interfaces: a hexadecane-water interface and a tricaprylin-water interface. The results show that, upon adsorption to both oil-water interfaces, β-Lg went through a β-sheet to α-helix transition with a corresponding loss of its globular tertiary structure. The degree of conformational change was also a function of the oil phase polarity. The hexadecane oil induced a much higher degree of non-native α-helix compared to the tricaprylin oil. In contrast to the β-Lg conformation in solution, the non-native α-helical-rich conformation of β-Lg at the interface was resistant to further conformational change upon heating. DPI measurements suggest that β-Lg formed a thin dense layer at emulsion droplet surfaces. The effects of high temperature and the presence of salt on these β-Lg emulsions were then investigated by monitoring changes in the ζ-potential and particle size. In the absence of salt, high electrostatic repulsion meant β-Lg-stabilized emulsions were resistant to heating to 90 °C. Adding salt (120 mM NaCl) before or after heating led to emulsion flocculation due to the screening of the electrostatic repulsion between colloidal particles. This study has provided insight into the structural properties of proteins adsorbed at the oil-water interface and has implications in the formulation and production of emulsions stabilized by globular proteins.     

Unified model to predict self-assembly of nonionic surfactants in solution and adsorption on solid or fluid hydrophobic surfaces: effect of molecular structure

We have developed a pseudo-phase model to predict the self-assembly of nonionic surfactants on hydrophobic solid or fluid interfaces and in bulk solution. The uniqueness of this model is that it provides the relationship between molecular structure and self-assembly in solution and on interfaces. This model requires the input of minimal new experimental data. The remaining model parameters may be calculated on the basis of the surfactant molecular structure. The validity of the model has been established by comparing predictions with a wide array of experimental data for nonionic surfactant adsorption at the hydrophobic solid-water interface and at the air-water interface. The same model is then used to predict the self-assembly in bulk solution. The model predictions for critical aggregation concentration, aggregate shapes, and adsorption isotherms of various surfactants are in good agreement with the experimental data available in the literature.     

Dynamics of protein and mixed protein/surfactant adsorption layers at the water/fluid interface

The adsorption behaviour of proteins and systems mixed with surfactants of different nature is described. In the absence of surfactants the proteins mainly adsorb in a diffusion controlled manner. Due to lack of quantitative models the experimental results are discussed partly qualitatively. There are different types of interaction between proteins and surfactant molecules. These interactions lead to protein/surfactant complexes the surface activity and conformation of which are different from those of the pure protein. Complexes formed with ionic surfactants via electrostatic interaction have usually a higher surface activity, which becomes evident from the more than additive surface pressure increase. The presence of only small amounts of ionic surfactants can significantly modify the structure of adsorbed proteins. With increasing amounts of ionic surfactants, however, an opposite effect is reached as due to hydrophobic interaction and the complexes become less surface active and can be displaced from the interface due to competitive adsorption. In the presence of non-ionic surfactants the adsorption layer is mainly formed by competitive adsorption between the compounds and the only interaction is of hydrophobic nature. Such complexes are typically less surface active than the pure protein. From a certain surfactant concentration of the interface is covered almost exclusively by the non-ionic surfactant. Mixed layers of proteins and lipids formed by penetration at the water/air or by competitive adsorption at the water/chloroform interface are formed such that at a certain pressure the components start to separate. Using Brewster angle microscopy in penetration experiments of proteins into lipid monolayers this interfacial separation can be visualised. A brief comparison of the protein adsorption at the water/air and water/n-tetradecane shows that the adsorbed amount at the water/oil interface is much stronger and the change in interfacial tension much larger than at the water/air interface. Also some experimental data on the dilational elasticity of proteins at both interfaces measured by a transient relaxation technique are discussed on the basis of the derived thermodynamic model. As a fast developing field of application the use of surface tensiometry and rheometry of mixed protein/surfactant mixed layers is demonstrated as a new tool in the diagnostics of various diseases and for monitoring the progress of therapies.     

Protein adsorption at the electrified air-water interface: implications on foam stability

The surface chemistry of ions, water molecules, and proteins as well as their ability to form stable networks in foams can influence and control macroscopic properties such as taste and texture of dairy products considerably. Despite the significant relevance of protein adsorption at liquid interfaces, a molecular level understanding on the arrangement of proteins at interfaces and their interactions has been elusive. Therefore, we have addressed the adsorption of the model protein bovine serum albumin (BSA) at the air-water interface with vibrational sum-frequency generation (SFG) and ellipsometry. SFG provides specific information on the composition and average orientation of molecules at interfaces, while complementary information on the thickness of the adsorbed layer can be obtained with ellipsometry. Adsorption of charged BSA proteins at the water surface leads to an electrified interface, pH dependent charging, and electric field-induced polar ordering of interfacial H(2)O and BSA. Varying the bulk pH of protein solutions changes the intensities of the protein related vibrational bands substantially, while dramatic changes in vibrational bands of interfacial H(2)O are simultaneously observed. These observations have allowed us to determine the isoelectric point of BSA directly at the electrolyte-air interface for the first time. BSA covered air-water interfaces with a pH near the isoelectric point form an amorphous network of possibly agglomerated BSA proteins. Finally, we provide a direct correlation of the molecular structure of BSA interfaces with foam stability and new information on the link between microscopic properties of BSA at water surfaces and macroscopic properties such as the stability of protein foams.     

Interfacial properties of heat-treated ovalbumin

The interfacial properties (kinetics of adsorption at the air/water interface, rheology of the interfacial layer) of ovalbumin molecules, unheated or previously heat-denatured in solution (10 g L(-1), pH 7, NaCl 50 mM) under controlled conditions (up to 40 min at 80 degrees C), were investigated. Heat treatments induced the formation of covalent aggregates which surface exhibits a higher hydrophobicity and an increased exposition of sulfhydryl groups when compared to native ovalbumin (unheated). Although they have a larger hydrodynamic size, aggregates adsorb as fast as native ovalbumin at the air/water interface. However, aggregates are able to established rapid contacts in the interfacial layer as shown by the fast increase of both surface pressure and shear elastic constant. In contrast, native ovalbumin needs longer time to developed intermolecular contacts and exhibits lower foam stability even if the shear elastic constant on aging reached higher value than for ovalbumin aggregates.