Julia Maldonado-Valderrama

Interfacial Characterization of β-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.

Effect of Gastric Conditions on β-Lactoglobulin Interfacial Networks: Influence of the Oil Phase on Protein Structure

Understanding the effects of digestion conditions on the structure of interfacial protein networks is important in order to rationally design food emulsions which can moderate lipid digestion. This study compares the effect of gastric conditions (pH, temperature, and ionic strength) on β-lactoglobulin films at different fluid interfaces: air-water, tetradecane-water, and olive oil-water. The experiments have been designed to simulate the passage into the stomach media. Hence, preformed interfacial protein (β-lactoglobulin) networks have been exposed to gastric conditions in order to establish generic aspects of the digestion process. The results show that the presence of an oil phase affects both the unfolding of the protein at the interface on adsorption and the subsequent interprotein associations responsible for network formation at the interface. Furthermore, the effects of the physiological conditions characteristic of the stomach also altered differently the preformed protein layer at different fluid interfaces. Initially, the effects of temperature, acid pH, and ionic strength on the dilatational modulus of β-lactoglobulin adsorbed layers at tetradecane-water and olive oil-water interfaces were studied in isolation. The presence of salt was found to have a major effect on the dilatational response at the oil-water interface in contrast to the observations at the air-water interface: it enhanced intermolecular association, hence increasing the packing at the interface causing it to become more elastic. Exposure to acid pH (2.5) also increased the elasticity of the interface, possibly due to the fact that strong electrostatic interactions acting at the interface compensated for the reduced level of intermolecular association. However, the increase in dilatational modulus at the oil-water interface was less noticeable upon exposure to combined changes in acid pH and ionic strength, as would occur in the stomach. This is consistent with previously reported observations at the air-water interface. The quantitative differences in the response of the protein networks to gastric media at different fluid interfaces are discussed in terms of the conformation of β-lactoglobulin within the networks formed at each interface based on detailed theoretical modeling of adsorption data.

The effect of physiological conditions on the surface structure of proteins: Setting the scene for human digestion of emulsions

Understanding and manipulating the interfacial mechanisms that control human digestion of food emulsions is a crucial step towards improved control of dietary intake. This article reports initial studies on the effects of the physiological conditions within the stomach on the properties of the film formed by the milk protein (

Interactions between Pluronics (F127 and F68) and Bile Salts (NaTDC) in the Aqueous Phase and the Interface of Oil-in-Water Emulsions

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Probing the in Situ Competitive Displacement of Protein by Nonionic Surfactant Using Atomic Force Microscopy

-lactoglobulin) at the air-water interface. Atomic force microscopy (AFM), surface tension and surface rheology techniques were used to visualize and examine the effect of gastric conditions on the network structure. The effects of changes in temperature, pH and ionic strength on a pre-formed interfacial structure were characterized in order to simulate the actual digestion process. Changes in ionic strength had little effect on the surface properties. In isolation, acidification reduced both the dilatational and the surface shear modulus, mainly due to strong repulsive electrostatic interactions within the surface layer and raising the temperature to body temperature accelerated the rearrangements within the surface layer, resulting in a decrease of the dilatational response and an increase of surface pressure. Together pH and temperature display an unexpected synergism, independent of the ionic strength. Thus, exposure of a pre-formed interfacial

On the Difference between Foams Stabilized by Surfactants and Whole Casein or β-Casein. Comparison of Foams, Foam Films, and Liquid Surfaces Studies

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Surface Characterization and AFM Imaging of Mixed Fibrinogen−Surfactant Films

-lactoglobulin film to simulated gastric conditions reduced the surface dilatational modulus and surface shear moduli. This is attributed to a weakening of the surface network in which the surface rearrangements of the protein prior to exposure to gastric conditions might play a crucial role.

Investigating the effect of an arterial hypertension drug on the structural properties of plasma protein

Pluronics are being introduced in food research in order to delay lipid digestion, with the length of hydrophilic and hydrophobic chains playing an important role in the rate of such a process. Since bile salts play a crucial role in the lipid digestion process, the aim of this work is to analyze the interactions between Pluronic F127 or F68 and the bile salt NaTDC when the latter is added at physiological concentrations. These interactions are studied at the Pluronic-covered oil-water interface and in the aqueous phase of Pluronic-stabilized emulsions. This work has been carried out with techniques such as differential scanning calorimetry, interfacial tension, dilatational rheology, and scanning electron microscopy. As a result, Pluronic F127 was shown to be more resistant to displacement by bile salt than F68 at the oil-water interface due to the larger steric hindrance and interfacial coverage provided. In addition, Pluronics have the ability to compete for the oil-water interface and interact in the bulk with the bile salt. Concretely, Pluronic F127 seems to interact with more molecules of bile salt in the bulk, thus hindering their adsorption onto the oil-water interface. As a conclusion, Pluronic F127 affects to a larger extent the ability of bile salt to promote the further cascade of lipolysis in the presence of lipase owing to a combination of interfacial and bulk events.

Atomic force microscopy as a tool to study the adsorption of DNA onto lipid interfaces

Force-distance data obtained from an atomic force microscope have been used to follow the in situ displacement of beta-lactoglobulin from tetradecane droplets by Tween 20 (polyoxyethylenesorbitan monolaurate). Interpretation of the force-distance curves has shown that the slope of the region, traditionally termed the constant compliance region, is a useful indicator of droplet deformation within a given experiment. The magnitude of this slope can be used to monitor how the deformability of the droplet changes upon addition of surfactant. It has been found that, immediately after initial addition of surfactant, there is an increase in magnitude of this slope, indicating a stiffening of the droplet, attributed to a stiffening of the protein network formed at the surface of the droplet. Subsequent additions of Tween 20 reduce the magnitude of the slope until an equilibrium value is reached, where the interface becomes surfactant-dominated. These observations suggest that it is possible to monitor in situ the displacement of protein from individual oil droplets. The data have been interpreted in terms of the orogenic model of displacement, which is based on studies made on model interfaces. These data have been compared to those obtained using the more traditional techniques of dilatational rheology, surface loading, and surface potential measurements for analogous beta-lactoglobulin-stabilized droplets or emulsions.

Interaction of surfactant and protein at the O/W interface and its effect on colloidal and biological properties of polymeric nanocarriers

This research work aims to investigate the behavior of a mixed system composed of a commercial protein (whole casein) and a low molecular weight surfactant (Tween 20) in order to understand its foam stability on the basis of fundamental surface quantities such as surface and disjoining pressure. These experiments prove to be extremely useful in the understanding of the processes determining foam stability. The complex behavior of whole casein/Tween 20 mixtures is directly deduced from the surface pressure isotherms. Concretely, the isotherm of the mixed system is displaced to smaller surfactant concentrations as compared to the pure surfactant system. This feature is quantified by a critical aggregation concentration and suggests formation of protein/surfactant complexes within the surface layer. The disjoining pressure isotherms of the pure and mixed systems provide key information regarding the structure and composition of the mixed whole casein/Tween 20 surface layers. Furthermore, they provide a direct correlation with the foam stability in terms of the thickness of the final foam film; the thinner the film, the less stable the foam. The experimental results are further discussed in terms of literature studies of similar systems, and a final rather accurate description of the system arises. This work investigates the importance of the nature of the protein in the stability of the foams of protein/surfactant mixtures and highlights the fundamental role of the surface properties in the understanding of such a major phenomenon in colloid science and technology.