Evaluation of Changes in Protein Quality of High-Pressure Treated Aqueous Aquafaba

Chickpea cooking water (CCW), known as aquafaba, has potential as a replacement for egg whites due to its emulsion and foaming properties which come from the proteins and starch that leach out from chickpeas into the cooking water. High pressure (HP) processing has the ability to modify the functional characteristics of proteins. It is hypothesized that HP processing could favorably affect the functional properties of CCW proteins by influencing their structure. The objective of this study to evaluate the effect of HP treatment on the associated secondary structure, emulsion properties and thermal characteristics of CCW proteins. A central composite rotatable design is used with pressure level (227–573 MPa) and treatment time (6–24 min) as HP variables, and concentration of freeze dried CCW aquafaba powder (11–29%) as product variable, and compared to untreated CCW powder. HP improves aquafaba emulsion properties compared to control sample. HP reduces protein aggregates by 33.3%, while β-sheets decreases by 4.2–87.6% in which both correlated to increasing protein digestibility. α-helices drops by 50%. It affects the intensity of some HP treated samples, but not the trend of bands in most of them. HP treatment decreases T_d and enthalpy because of increasing the degree of denaturation.     

Effects of High-Intensity Ultrasound Pretreatment on Structure,Properties, and Enzymolysis of Walnut Protein Isolate

The purpose of this paper was to investigate the effect of high-intensity ultrasonication (HIU) pretreatment before enzymolysis on structural conformations of walnut protein isolate (WPI) and antioxidant activity of its hydrolysates. Aqueous WPI suspensions were subjected to ultrasonic processing at different power levels (600–2000 W) and times (5–30 min), and then changes in the particle size, zeta (ζ) potential, and structure of WPI were investigated, and antioxidant activity of its hydrolysates was determined. The particle size of the particles of aqueous WPI suspensions was decreased after ultrasound, indicating that sonication destroyed protein aggregates. The ζ-potential values of a protein solution significantly changed after sonication, demonstrating that the original dense structure of the protein was destroyed. Fourier transform infrared spectroscopy indicated a change in the secondary structure of WPI after sonication, with a decrease in β-turn and an increase in α-helix, β-sheet, and random coil content. Two absorption peaks of WPI were generated, and the fluorescence emission intensity of the proteins decreased after ultrasonic treatment, indicating that the changes in protein tertiary structure occurred. Moreover, the degree of hydrolysis and the antioxidant activity of the WPI hydrolysates increased after sonication. These results suggest that HIU pretreatment is a potential tool for improving the functional properties of walnut proteins.     

Protein interactions and association: an open challenge for colloid science

The solution behaviour of globular proteins still presents many puzzling aspects, calling for a deeper understanding of the physical properties of lyophilic colloids. For instance, protein interactions in ‘salting-out’ conditions require to take explicitly into account ‘Donnan’ effects on the small ion distribution, while understanding of temperature and salt–specificity effects on protein solubility presumably calls for a careful analysis of hydrophobic contributions. Yet, very unexpected effects can be found even at low salt concentration. In particular, we discuss the very distinctive properties of β-lactoglobulin A (BLGA) solutions, where strong attractive interactions show up, displaying a marked non-monotonic trend as a function of the solution ionic strength. These ‘anomalous’ attractions drive very peculiar reversible association processes, resulting in the spontaneous formation of short-lived clusters with a well-defined small aggregation number. We suggest that other protein association processes of physiological interest may parallel BLGA clustering.     

Rheological and Solubility Properties of Soy Protein Isolate

Soy protein isolate (SPI) powders often have poor water solubility, particularly at pH values close to neutral, which is an attribute that is an issue for its incorporation into complex nutritional systems. Therefore, the objective of this study was to improve SPI solubility while maintaining low viscosity. Thus, the intention was to examine the solubility and rheological properties of a commercial SPI powder at pH values of 2.0, 6.9, and 9.0, and determine if heat treatment at acidic or alkaline conditions might positively influence protein solubility, once re-adjusted back to pH 6.9. Adjusting the pH of SPI dispersions from pH 6.9 to 2.0 or 9.0 led to an increase in protein solubility with a concomitant increase in viscosity at 20 °C. Meanwhile, heat treatment at 90 °C significantly improved the solubility at all pH values and resulted in a decrease in viscosity in samples heated at pH 9.0. All SPI dispersions measured under low-amplitude rheological conditions showed elastic-like behaviour (i.e., G′ > G″), indicating a weak “gel-like” structure at frequencies less than 10 Hz. In summary, the physical properties of SPI can be manipulated through heat treatment under acidic or alkaline conditions when the protein subunits are dissociated, before re-adjusting to pH 6.9.     

Physicochemical and Functional Properties of 2S, 7S,and 11S Enriched Hemp Seed Protein Fractions

The hemp seed contains protein fractions that could serve as useful ingredients for food product development. However, utilization of hemp seed protein fractions in the food industry can only be successful if there is sufficient information on their levels and functional properties. Therefore, this work provides a comparative evaluation of the structural and functional properties of hemp seed protein isolate (HPI) and fractions that contain 2S, 7S, or 11S proteins. HPI and protein fractions were isolated at pH values of least solubility. Results showed that the dominant protein was 11S, with a yield of 72.70 ± 2.30%, while 7S and 2S had values of 1.29 ± 0.11% and 3.92 ± 0.15%, respectively. The 2S contained significantly (p < 0.05) higher contents of sulfhydryl groups at 3.69 µmol/g when compared to 7S (1.51 µmol/g), 11S (1.55 µmol/g), and HPI (1.97 µmol/g). The in vitro protein digestibility of the 2S (72.54 ± 0.52%) was significantly (p < 0.05) lower than those of the other isolated proteins. The intrinsic fluorescence showed that the 11S had a more rigid structure at pH 3.0, which was lost at higher pH values. We conclude that the 2S fraction has superior solubility, foaming capacity, and emulsifying activity when compared to the 7S, 11S, and HPI.     

Adsorption at the air-water interface and emulsification properties of grain legume protein derivatives from pea and broad bean

Functional properties of native and modified (through induced autolysis) pea (Pisum sativum L.) and broad bean (Vicia faba L.) protein derivatives are studied. In specific, protein solubility and behavior at the air–water interface through surface pressure measurements are investigated. Furthermore the ability of the protein products to act as emulsifying agents and to stabilize emulsions is studied through oil droplet size distribution measurements and by the protein adsorbed at the oil–water interface. The data reveal that the ability of the proteins to act as surfactants and build up a rigid film around the oil droplets, mainly depends on their suitable molecular configuration and structure. Hydrolysis did not promote the functionality of the legume proteins. Broad bean exhibited better functionality than pea, before and after hydrolysis. Some comparisons were also made with lupin (Lupinus albus L.) protein isolate.     

The effect of sample salt additives on capillary electrophoresis analysis of intact proteins using surface modified capillaries

The effect of adding alkali salts to protein samples for capillary electrophoretic (CE) analysis of intact proteins was studied. A high degree of peak stacking, even for large proteins, was found to occur when alkali salts were added to the sample. The addition of salt to the protein sample promotes a strong improvement in the peak efficiency of individual proteins giving up to 2.1 × 10⁶ apparent plates/m. The concentration of salt required in the sample to reach optimal peak efficiency show dependency on both the molecular weight and molar concentration of the protein. However, adding salt will, at a sufficiently high concentration, cause a mixture of proteins to co-migrate to one very sharp peak. The observed sample stacking effect was obtained with a number of different surface modified silica capillaries indicating a general phenomenon and not surface coating specific.     

Experimental Determination and Computational Prediction of Dehydroabietic Acid Solubility in (−)-α-Pinene + (−)-β-Caryophyllene + P-Cymene System

The solubility of dehydroabietic acid in (−)-α-pinene, p-cymene, (−)-β-caryophyllene, (−)-α-pinene + p-cymene, (−)-β-caryophyllene + p-cymene and (−)-α-pinene + (−)-β-caryophyllene were determined using the laser monitoring method at atmospheric pressure. The solubility of dehydroabietic acid was positively correlated with temperature from 295.15 to 339.46 K. (−)-α-pinene, p-cymene, and (−)-β-caryophyllene were found to be suitable for the solubilization of dehydroabietic acid. In addition, the non-random two liquid (NRTL), universal quasi-chemical (UNIQUAC), modified Apelblat, modified Wilson, modified Wilson–van’t Hoff, and λh models were applied to correlate the determined solubility data. The modified Apelblat model gave the minor deviation for dehydroabietic acid in monosolvents, while the λh equation showed the best result in the binary solvents. A comparative analysis of compatibility between solutes and solvents was carried out using Hansen solubility parameters. The thermodynamic functions of Δ_solH⁰, Δ_solS⁰, Δ_solG⁰ were calculated according to the van’t Hoff equation, indicating that the dissolution was an entropy-driven heat absorption process. The Conductor-like Screening Model for Real Solvents (COSMO-RS) combined with an experimental value was applied to predict the reasonable solubility data of dehydroabietic acid in the selected solvents systems. The interaction energy of the dehydroabietic acid with the solvent was analyzed by COSMO-RS.     

Characterization of Conjugates between α-Lactalbumin and Benzyl Isothiocyanate—Effects on Molecular Structure and Proteolytic Stability

In complex foods, bioactive secondary plant metabolites (SPM) can bind to food proteins. Especially when being covalently bound, such modifications can alter the structure and, thus, the functional and biological properties of the proteins. Additionally, the bioactivity of the SPM can be affected as well. Consequently, knowledge of the influence of chemical modifications on these properties is particularly important for food processing, food safety, and nutritional physiology. As a model, the molecular structure of conjugates between the bioactive metabolite benzyl isothiocyanate (BITC, a hydrolysis product of the glucosinolate glucotropaeolin) and the whey protein α-lactalbumin (α-LA) was investigated using circular dichroism spectroscopy, anilino-1-naphthalenesulfonic acid fluorescence, and dynamic light scattering. Free amino groups were determined before and after the BITC conjugation. Finally, mass spectrometric analysis of the BITC-α-LA protein hydrolysates was performed. As a result of the chemical modifications, a change in the secondary structure of α-LA and an increase in surface hydrophobicity and hydrodynamic radii were documented. BITC modification at the ε-amino group of certain lysine side chains inhibited tryptic hydrolysis. Furthermore, two BITC-modified amino acids were identified, located at two lysine side chains (K32 and K113) in the amino acid sequence of α-LA.     

Competitive adsorption of -α-lysophosphatidylcholine/β-lactoglobulin mixtures at the interfaces of foams and foam lamellae

The stability of foams formed with the protein β-lactoglobulin as a function of increasing concentration of the lipid analogue -α-lysophosphatidylcholine were investigated using a microconductivity technique. The drainage, surface diffusion and thickness properties of thin liquid films (foam lamallae) were also studied using optical microscopy including epi-illumination, fluorescence recovery after photobleaching and film interferometry techniques. In addition, the surfactant binding properties of the protein were examined. The addition of small quantities of -α-lysophosphatidylcholine to β-lactoglobulin (molar ratio, R < 7:1) increased the foam stability, whereas a slightly higher concentration of surfactant in the mixture (R = 10) caused foam destabilisation. The explanation of these observations is based on changes in the composition and structure of the adsorbed interfacial layers of the thin films caused by competitive displacement of the protein by the surfactant.     

Hydrophobicity Enhances the Formation of Protein-Stabilized Foams

Screening proteins for their potential use in foam applications is very laborious and time consuming. It would be beneficial if the foam properties could be predicted based on their molecular properties, but this is currently not possible. For protein-stabilized emulsions, a model was recently introduced to predict the emulsion properties from the protein molecular properties. Since the fundamental mechanisms for foam and emulsion formation are very similar, it is of interest to determine whether the link to molecular properties defined in that model is also applicable to foams. This study aims to link the exposed hydrophobicity with the foam ability and foam stability, using lysozyme variants with altered hydrophobicity, obtained from controlled heat treatment (77 °C for 0–120 min). To establish this link, the molecular characteristics, interfacial properties, and foam ability and stability (at different concentrations) were analysed. The increasing hydrophobicity resulted in an increased adsorption rate constant, and for concentrations in the protein-poor regime, the increasing hydrophobicity enhanced foam ability (i.e., interfacial area created). At higher relative exposed hydrophobicity (i.e., ~2–5 times higher than native lysozyme), the adsorption rate constant and foam ability became independent of hydrophobicity. The foam stability (i.e., foam collapse) was affected by the initial foam structure. In the protein-rich regime—with nearly identical foam structure—the hydrophobicity did not affect the foam stability. The link between exposed hydrophobicity and foam ability confirms the similarity between protein-stabilized foams and emulsions, and thereby indicates that the model proposed for emulsions can be used to predict foam properties in the future.     

Effects of pH and Ionic Salts on the Emulsifying and Rheological Properties of Acorn Protein Isolate

This study was designed to evaluate the emulsifying and rheological properties of acorn protein isolate (API) in different pH mediums (pH 3, 7 and 9) and in the presence of ionic salts (1 M NaCl and 1 M CaCl₂). API shows higher solubility in distilled water at pH 7, while at the same pH, a decrease in solubility was observed for API in the presence of CaCl₂ (61.30%). A lower emulsifying activity index (EAI), lower stability index (ESI), larger droplet sizes and slight flocculation were observed for API in the presence of salts at different pHs. Importantly, CaCl₂ treated samples showed relevantly higher EAI (252.67 m²/g) and ESI (152.67 min) values at all pH as compared to NaCl (221.76 m²/g), (111.82 min), respectively. A significant increase in interfacial protein concentration (4.61 mg/m²) was observed for emulsion at pH 9 with CaCl₂, while the major fractions of API were observed in an interfacial layer after SDS-PAGE analysis. All of the emulsion shows shear thinning behavior (τ_c > 0 and n < 1), while the highest viscosity was observed for emulsion prepared with CaCl₂ at pH 3 (11.03 ± 1.62). In conclusion, API, in the presence of ionic salts at acidic, neutral and basic pH, can produce natural emulsions, which could be substitutes for synthetic surfactants for such formulations.     

Effect of protein aggregates on foaming properties of β-lactoglobulin

Our paper aims at determining the respective part of protein aggregates and non-aggregated proteins in the foam formation and stability of β-lactoglobulin. We report results on fractal aggregates formed at neutral pH and strong ionic strength (aggregates size from 30 to 190 nm). Pure aggregates and mixtures of non-aggregated/aggregated proteins at varying ratios were used. The capacity of aggregates to form and stabilize foams has been studied in relation with their ability to absorb at air/water interfaces. Our results show that protein aggregates are not able by themselves to improve the foaming properties but participate to a better foam stabilization in the presence of non-aggregated proteins. Non-aggregated proteins appear to be necessary to produce stable foams. We have shown that the amount and the size of aggregates had an influence on the drainage rate.     

Ionic Conduction of Solid Polymer Electrolytes Based on Acrylonitrile–Butadiene Copolymer and Lithium Hexafluoroarsenate

The ionic structure and transport properties intrinsic to solid polymer electrolytes based on an acrylonitrile–butadiene (40 : 60) copolymer and lithium hexafluoroarsenate are studied at the salt concentrations close to the salt solubility limit in the polymer matrix. The ionic structure of a macromolecular ionic solution alters with increasing salt concentration and temperature, as does the ionic-transport mechanism. Possible models for the ionic structure of concentrated solid polymer electrolytes and ionic-transport mechanisms corresponding to them are considered.     

Optimization of the Physical,Optical and Mechanical Properties of Composite Edible Films of Gelatin,Whey Protein and Chitosan

The aim of this work was to evaluate the effect of the concentration of gelatin (G) (3–6 g), whey protein (W) (2.5–7.5 g) and chitosan (C) (0.5–2.5 g) on the physical, optical and mechanical properties of composite edible films (CEFs) using the response surface methodology (RSM), as well as optimizing the formulation for the packaging of foods. The results of the study were evaluated via first- and second-order multiple regression analysis to obtain the determination coefficient values with a good fit (R ˃ 0.90) for each of the response variables, except for the values of solubility and b*. The individual linear effect of the independent variables (the concentrations of gelatin, whey protein and chitosan) significantly affected (p ≤ 0.05) the water vapor permeability (WVP), strength and solubility of the edible films. The WVP of the edible films varied from 0.90 to 1.62 × 10⁻¹¹ g.m/Pa.s.m², the resistance to traction varied from 0.47 MPa to 3.03 MPa and the solubility varied from 51.06% to 87%. The optimized values indicated that the CEF prepared with a quantity of 4 g, 5 g and 3 g of gelatin, whey protein and chitosan, respectively, provided the CEF with a smooth, continuous and transparent surface, with L values that resulted in a light-yellow hue, a lower WVP, a maximum strength (resistance to traction) and a lower solubility. The results revealed that the optimized formulation of the CEF of G–W–C allowed a good validation of the prediction model and could be applied, in an effective manner, to the food packaging industry, which could help in mitigating the environmental issues associated with synthetic packaging materials.     

FT-Raman Spectroscopy as a Tool to Study the Secondary Structures of Wheat Gliadin Proteins

Raman spectroscopy is a useful method in biological, biomedical, food, and agricultural studies, allowing the simultaneous examination of various chemical compounds and evaluation of molecular changes occurring in tested objects. The purpose of our research was to explain how the elimination of ω-fractions from the wheat gliadin complex influences the secondary structures of the remaining αβγ-gliadins. To this aim, we analyzed the endosperm of wheat kernels as well as gliadin proteins extracted from two winter wheat genotypes: wasko.gl+ (control genotype containing the full set of gliadins) and wasko.gl− (modified genotype lacking all ω-gliadins). Based on the decomposition of the amide I band, we observed a moderate increase in β-forms (sheets and turns) at the expense of α-helical and random coil structures for gliadins isolated from the flour of the wasko.gl− line. Since ω-gliadins contain no cysteine residues, they do not participate in the formation of the disulfide bridges that stabilize the protein structure. However, they can interact with other proteins via weak, low-energetic hydrogen bonds. We conclude that the elimination of ω-fractions from the gliadin complex causes minor modifications in secondary structures of the remaining gliadin proteins. In our opinion, these small, structural changes of proteins may lead to alterations in gliadin allergenicity.     

Emulsification and Foaming Properties of Hydrophobically Modified Gelatin

Surface active gelatins were formed by covalent attachment of hydrophobic groups to gelatin molecules by reactingN-hydroxysuccinimide esters of various fatty acids (C₄–C₁₆) with the lysine groups. The surface activity was evaluated by emulsification and foaming properties, and by adsorption at the oil–water interface. It was found that, in general, the modified gelatins are more surface active than the native gelatin. The increase in hydrophobic chain length and the number of attached alkyl chains per gelatin molecule leads to a decrease in the emulsion droplet's size and to more stable emulsions. Adsorption isotherms, at the o/w interface, show much higher surface concentration, at saturation, of the modified gelatin than the native gelatin. The modified gelatins also have high foaming ability and a high foam stability, while the maximal foam activity is obtained by the C₈modified gelatin. The foaming properties of the surface-active gelatins were also compared to that of sodium dodecyl sulfate (SDS) and it was found that below the CMC of SDS, both foam activity and stability were higher for the modified gelatins. On the other hand, above the CMC the foam activity of SDS was higher, but the foam stability was lower than for C₈–C₁₆-modified gelatins.     

Effects of Hexane on Protein Profile,Solubility and Foaming Properties of Defatted Proteins Extracted from Tenebrio molitor Larvae

Inclusion of edible insects in human diets is increasingly promoted as a sustainable source of proteins with high nutritional value. While consumer acceptability remains the main challenge to their integration into Western food culture, the use of edible insects as meal and protein concentrate could decrease neophobia. The defatting of edible insects, mostly done with hexane, is the first step in producing protein ingredients. However, its impact on protein profiles and techno-functionality is still unclear. Consequently, this study compares the protein profiles of hexane-defatted and non-hexane-defatted yellow mealworm (Tenebrio molitor) meals and protein extracts, and evaluates the impact of hexane on protein solubility and foaming properties. Results showed that profiles for major proteins were similar between hexane-defatted and non-defatted samples, however some specific content differences (e.g., hexamerin 2) were observed and characterized using proteomic tools. Protein solubility was markedly lower for T. molitor meals compared to protein extracts. A large increase in the foaming capacity was observed for defatted fractions, whereas foam stability decreased similarly in all fractions. Consequently, although the hexane-defatting step was largely studied to produce edible insect protein ingredients, it is necessary to precisely understand its impact on their techno-functional properties for the development of food formulations.