Unravelling the Influence of Composition and Heat Treatment on Key Characteristics of Dairy Protein Powders Using a Multifactorial Approach.

The purpose of this study was to improve understanding of the structural and functional property changes that milk-protein concentrates undergo during production, particularly how the manufacturing route (heat treatment position and intensity), standardization (in osmosed water or ultrafiltrate permeate) and formulation (casein:whey protein (Cas:WP) ratio) influence the physico-chemical characteristics-hygroscopicity, particle size, sphericity, density and evolution of browning during storage. To obtain a comprehensive understanding of the parameters responsible for the distinctive characteristics of different powders, a multifactorial approach was adopted. Hygroscopicity depended mainly on the standardizing solution and to a lesser extent the Cas:WP ratio. The particle size of the heat-treated casein-dominant powders was up to 5 µm higher than for those that had had no heat treatment regardless of the standardizing solution, which also had no influence on the sphericity of the powder particles. The density of the powders increased up to 800 kg·m-3 with a reduced proportion of casein, and lactose and whey proteins participated in browning reactions during storage at 13 °C. In increasing order, the modality of heat treatment, the standardizing solution and the Cas:WP protein ratio influenced the key characteristics. This work is relevant for industrial applications to increase control over the functionalities of powdered products.

Heat treatment of milk protein concentrates affects enzymatic coagulation properties.

Dairy ingredients with highly concentrated protein contents are high added value products with expanding market. The manufacture of such ingredients includes a succession of unit operations of which heat treatment is a key step to guarantee the microbial safety, that induces major changes in protein structures and thus ingredients functionalities. However, due to an incomplete understanding of phenomena taking place at high protein concentrations, shedding light on their mechanisms is a scientific challenge as well as an industrial need. In this study, the influence of heat treatment (74 °C/ 30 s) of highly concentrated milk protein systems (up to 20 % w/w) on protein denaturation/aggregation and enzymatic coagulation properties was studied using an original semi-industrial approach. 10 % w/w protein solutions constituted with whey protein and casein micelles at milk ratio, standardized in osmosed water or ultrafiltration permeate were used. These protein solutions were processed in different ways prior the manufacture of powders: heat treatment of the 10 % w/w protein solution before vacuum evaporation, heat treatment of the 20 % w/w protein solution after vacuum concentration, two consecutive heat treatments before and after vacuum evaporation. A fourth powder was prepared from unheated 10 % w/w protein solution. An increase in protein concentration led to a higher heat-induced protein denaturation. This phenomenon was reduced when increasing the lactose content. The effect of heat treatment on the extent of protein denaturation was not cumulative. At high protein concentration, interactions between κ-casein and whey protein were modified compared to milk, as mainly micelle-bound aggregates were formed at pH about 6.7. This phenomenon was enhanced at low ionic strength and lactose content. Our study showed that the enzymatic coagulation properties of reconstituted protein powders could be correlated with their physico-chemical compositions. An increase in protein denaturation disrupted the gel reorganization and led to the formation of weaker gels but did not interfere on the micelles aggregation phase and the early gelation. On the contrary, an increase in ionic strength and lactose content led to higher gel time.

Phase Diagram of Dairy Protein Mixes Obtained by Single Droplet Drying Experiments.

Dairy powders are mainly produced by droplet spray drying, an articulated process that enables the manufacture of high added-value goods with a long shelf life and well-preserved functional properties. Despite the recent advances, a full understanding of the mechanisms occurring at the droplet scale in drying towers and, consequently, of the impact of process parameters and processed fluid characteristics on the powder properties is far from being achieved. In the wake of previous studies based on a laboratory scale approach, in this work, we provided a global picture of the drying in droplets of dairy protein mixes, i.e., whey proteins and casein micelles, which represent crucial dairy powder ingredients. Using profile visualization and optical microscopy, we explored the shape evolution in droplets with a range of protein contents and compositions typical of commercial powder production. The observation favored the evaluation of the specific role of each protein on the evaporation dynamics, and led to the construction of a phase diagram predictive of the dry droplet shape starting from the characteristics of the initial protein dispersions. Our outcomes represent a further step shedding light on the paradigm linking the physics of drying at the microscale and the nutritional properties of complex dairy powders.

Protein Transport upon Advection at the Air/Water Interface: When Charge Matters.

The formation of dense protein interfacial layers at a free air-water interface is known to result from both diffusion and advection. Furthermore, protein interactions in concentrated phases are strongly dependent on their overall positive or negative net charge, which is controlled by the solution pH. As a consequence, an interesting question is whether the presence of an advection flow of water toward the interface during protein adsorption produces different kinetics and interfacial structure of the adsorbed layer, depending on the net charge of the involved proteins and, possibly, on the sign of this charge. Here we test a combination of the following parameters using ovalbumin and lysozyme as model proteins: positive or negative net charge and the presence or absence of advection flow. The formation and the organization of the interfacial layers are studied by neutron reflectivity and null-ellipsometry measurements. We show that the combined effect of a positive charge of lysozyme and ovalbumin and the presence of advection flow does induce the formation of interfacial multilayers. Conversely, negatively charged ovalbumin forms monolayers, whether advection flow is present or not. We show that an advection/diffusion model cannot correctly describe the adsorption kinetics of multilayers, even in the hypothesis of a concentration-dependent diffusion coefficient as in colloidal filtration, for instance. Still, it is clear that advection is a necessary condition for making multilayers through a mechanism that remains to be determined, which paves the way for future research.

The solute mechanical properties impact on the drying of dairy and model colloidal systems.

The evaporation of colloidal solutions is frequently observed in nature and in everyday life. The investigation of the mechanisms taking place during the desiccation of biological fluids is currently a scientific challenge with potential biomedical and industrial applications. In the last few decades, seminal works have been performed mostly on dried droplets of saliva, urine and plasma. However, the full understanding of the drying process in biocolloids is far from being achieved and, notably, the impact of solute properties on the morphological characteristics of the evaporating droplets, such as colloid segregation, skin formation and crack pattern development, is still to be elucidated. For this purpose, the use of model colloidal solutions, whose rheological behavior is more easily deducible, could represent a significant boost. In this work, we compare the drying of droplets of whey proteins and casein micelles, the two main milk protein classes, to that of dispersions of silica particles and polymer-coated silica particles, respectively. The mechanical behavior of such biological colloids and model silica dispersions was investigated through the analysis of crack formation, and the measurements of their mechanical properties using indentation testing. The study reveals numerous analogies between dairy and the corresponding model systems, thus confirming the latter as a plausible powerful tool to highlight the signature of the matter at the molecular scale during the drying process.

Interfacial properties, film dynamics and bulk rheology: A multi-scale approach to dairy protein foams.

HYPOTHESIS: The effective contribution of interfacial properties to the rheology of foams is a source of many open questions. Film dynamics during topological T1 changes in foams, essentially studied for low molecular weight surfactants, and scarcely for proteins, could connect interfacial properties to protein foam rheology. EXPERIMENTS: We modified whey protein isolate (WPI), and its purified major protein ß-lactoglobulin (ß-lg) by powder pre-conditioning and dry-heating in order to obtain a broad variety of interfacial properties. We measured interfacial properties, film relaxation duration after a T1 event and bulk foam rheology. FINDINGS: We found that, for ß-lg, considered as a model protein, the higher the interfacial elastic modulus, the longer the duration of topological T1 changes and the greater the foam storage and loss moduli and the yield stress. However, in the case of the more complex WPI, these correlations were less clear. We propose that the presence in WPI of other proteins, lactose and minerals modify the impact of pre-conditioning and dry-heating on proteins and thereby, their behaviour at the interface and inside the liquid film.

Soft-Matter Approaches for Controlling Food Protein Interactions and Assembly.

Animal- and plant-based proteins are present in a wide variety of raw and processed foods. They play an important role in determining the final structure of food matrices. Food proteins are diverse in terms of their biological origin, molecular structure, and supramolecular assembly. This diversity has led to segmented experimental studies that typically focus on one or two proteins but hinder a more general understanding of food protein structuring as a whole. In this review, we propose a unified view of how soft-matter physics can be used to control food protein assembly. We discuss physical models from polymer and colloidal science that best describe and predict the phase behavior of proteins. We explore the occurrence of phase transitions along two axes: increasing protein concentration and increasing molecular attraction. This review provides new perspectives on the link between the interactions, phase transitions, and assembly of proteins that can help in designing new food products and innovative food processing operations.

Drying colloidal systems: Laboratory models for a wide range of applications.

The drying of complex fluids provides a powerful insight into phenomena that take place on time and length scales not normally accessible. An important feature of complex fluids, colloidal dispersions and polymer solutions is their high sensitivity to weak external actions. Thus, the drying of complex fluids involves a large number of physical and chemical processes. The scope of this review is the capacity to tune such systems to reproduce and explore specific properties in a physics laboratory. A wide variety of systems are presented, ranging from functional coatings, food science, cosmetology, medical diagnostics and forensics to geophysics and art.

Investigation of secondary structure evolution of micellar casein powder upon aging by FTIR and SRCD: consequences on solubility.

BACKGROUND: Synchrotron radiation circular dichroism (SRCD) and Fourier transform infrared (FTIR) spectroscopy were used to examine the conformation evolution of micellar casein (MC) powder during storage and to determine whether the spectral changes could be related to their solubility evolution. RESULTS: A loss in intensity of SRCD spectra as a function of storage time has been observed. Quantification of secondary structures revealed losses of α-helix content during storage. Moreover, a redshift of the amide I band in the FTIR spectrum was demonstrated during the storage and was interpreted as a rearrangement of the secondary structure of the protein, which is in line with the SRCD results. The qualitative results obtained by FTIR clearly support the quantitative evolution of the secondary structure obtained by the analysis of SRCD spectra. Principal component analysis (PCA) of FTIR spectra permits a good separation of samples according to the storage time. PCA shows that the evolution of secondary structures and solubility loss are closely linked. CONCLUSION: With the quantitative data provided by SRCD spectra, it was established that, whatever the storage conditions, a unique curve exists between loss of α-helix content and loss in solubility, showing that loss of α-helix content is a marker of solubility loss for the MC powders studied. © 2017 Society of Chemical Industry.

Osmotic pressures of lysozyme solutions from gas-like to crystal states.

We obtained osmotic pressure data of lysozyme solutions, describing their physical states over a wide concentration range, using osmotic stress for pressures between 0.05 bar and about 40 bar and volume fractions between 0.01 and 0.61. The osmotic pressure vs. volume fraction data consist of a dilute, gas-phase regime, a transition regime with a high-compressibility plateau, and a concentrated regime where the system is nearly incompressible. The first two regimes are shifted towards a higher protein volume fraction upon decreasing the strength or the range of electrostatic interactions. We describe this shift and the overall shape of the experimental data in these two regimes through a model accounting for a steric repulsion, a short-range van der Waals attraction and a screened electrostatic repulsion. The transition is caused by crystallization, as shown by small-angle X-ray scattering. We verified that our data points correspond to thermodynamic equilibria, and thus that they consist of the reference experimental counterpart of a thermodynamic equation of state.

Moderate conformational impact of citrate on ovotransferrin considerably increases its capacity to self-assemble at the interface.

We have compared the behavior of ovotransferrin at the air-solution interface in the presence of a monovalent ion (acetate), or a divalent ion (citrate), the latter being known to induce conformational changes of this protein upon interaction with its iron-binding sites. We have characterised the adsorption layer at the air-water interface in terms of homogeneity, surface concentration excess and rheological properties at pH 4.0. Besides we have investigated the bulk conformation in the presence of the two anions. In the presence of citrate only, interfacial layers display well-defined domains of higher overall surface concentration suggesting multilayers adsorption. Citrate also induces higher helical content and stabilizes the protein against thermal denaturation. Hence we propose that these changes are involved in the propensity of ovotransferrin to self-assemble at the air-water interface resulting in thick and heterogeneous interfacial layer.

Shape, shell, and vacuole formation during the drying of a single concentrated whey protein droplet.

The drying of milk concentrate droplets usually leads to specific particle morphology influencing their properties and their functionality. Understanding how the final shape of the particle is formed therefore represents a key issue for industrial applications. In this study, a new approach to the investigation of droplet-particle conversion is proposed. A single droplet of concentrated globular proteins extracted from milk was deposited onto a hydrophobic substrate and placed in a dry environment. Complementary methods (high-speed camera, confocal microscopy, and microbalance) were used to record the drying behavior of the concentrated protein droplets. Our results showed that whatever the initial concentration, particle formation included three dynamic stages clearly defined by the loss of mass and the evolution of the internal and external shapes of the droplet. A new and reproducible particle shape was related in this study. It was observed after drying a smooth, hemispherical cap-shaped particle, including a uniform protein shell and the nucleation of an internal vacuole. The particle morphology was strongly influenced by the drying environment, the contact angle, and the initial protein concentration, all of which governed the duration of the droplet shrinkage, the degree of buckling, and the shell thickness. These results are discussed in terms of specific protein behaviors in forming a predictable and a characteristic particle shape. The way the shell is formed may be the starting point in shaping particle distortion and thus represents a potential means of tuning the particle morphology.

Unexpected differences in the behavior of ovotransferrin at the air-water interface at pH 6.5 and 8.0.

Adsorption of purified apo-ovotransferrin at the air-water interface was studied by ellipsometry, surface tension, polarization-modulation infrared reflection-absorption spectroscopy (PM-IRRAS), and shear elastic constant measurements. No significant difference was observed between pH 6.5 and 8.0 as regards the final value of surface concentration and surface pressure. However at low concentration, a weak barrier to adsorption is evidenced at pH 6.5 and confirmed by PM-IRRAS measurements. At a pH where the protein net charge is negative (pH 8.0), the behavior of ovotransferrin at the air-water interface is more influenced by charge effects rather than bulk concentration effects. At this pH, the interface exhibits a low shear elastic constant and a spectral signature not usual for globular proteins.