Protein aggregation caused by pasteurization processing affects the foam performance of liquid egg white.

Pasteurization is necessary during the production of liquid egg whites (LEW), but the thermal effects in pasteurization could cause an unavoidable loss of foaming properties of LEW. This study intended to investigate the mechanism of pasteurization processing affects the foam performance of LEW. The foaming capacity (FC) of LEW deteriorated significantly (ΔFCmax = 72.33 %) and foaming stability (FS) increased slightly (ΔFSmax = 3.64 %) under different temperature-time combinations of pasteurization conditions (P < 0.05). The increased turbidity and the decreased solubility together with the decreased absolute value of Zeta potential indicated the generation of thermally induced aggregates and the instability of the protein particles, Rheological characterization demonstrated improved viscoelasticity in pasteurization liquid egg whites (PLEW), explaining enhanced FS. The study revealed that loss in foaming properties of PLEW resulted from thermal-induced protein structural changes and aggregation, particularly affecting FC. This provided a theoretical reference for the production and processing of LEW products.

Inhibition of water-diluted precipitate formation from egg whites by ultrasonic pretreatment: Insights from quantitative proteomics analysis.

The formation of the egg white precipitate (EWP) during dilution poses challenges in food processing. In this paper, the effects of 90 W and 360 W ultrasonic intensities on the inhibition of EWP formation were investigated. The findings revealed that 360 W sonication effectively disrupted protein aggregates, decreasing the dry matter of EWP by 5.24 %, particle size by 57.86 %, and viscosity by 82.28 %. Furthermore, the ultrasonic pretreatment unfolded protein structures and increased the content of ß-sheet structures. Combined with quantitative proteomics and intermolecular forces analysis, the mechanism by which ultrasonic pretreatment inhibited water-diluted EWP formation by altering protein interactions was proposed: ultrasonic pretreatment disrupted electrostatic interactions centered on lysozyme, as well as hydrogen-bonding interactions between ovomucin and water. In conclusion, our research provides valuable insights into the application of ultrasonic pretreatment as a means to control and improve the quality of egg white-based products.

Frequency dependence of ultrasonic effects on the kinetics of hen egg white lysozyme fibrillation.

Our study aimed to investigate the effects of ultrasound on the fibrillation kinetics of HEWL (hen egg white lysozyme) and its physicochemical properties. Ultrasound, a mechanical wave, can induce conformational changes in proteins. To achieve this, we developed an ultrasound exposure system and used various biophysical techniques, including ThT fluorescence spectroscopy, ATR-FTIR, Far-UV CD spectrophotometry, Fluorescence microscopy, UV-spectroscopy, and seeding experiments. Our results revealed that higher frequencies significantly accelerated the fibrillation of lysozyme by unfolding the native protein and promoting the fibrillation process, thereby reducing the lag time. We observed a change in the secondary structure of the sonicated protein change to the ß-structure, but there was no difference in the Tm of native and sonicated proteins. Furthermore, we found that higher ultrasound frequencies had a greater seeding effect. We propose that the effect of frequency can be explained by the impact of the Reynolds number, and for the Megahertz frequency range, we are almost at the transition regime of turbulence. Our results suggest that laminar flows may not induce any significant change in the fibrillation kinetics, while turbulent flows may affect the process.

Absorption of egg white hydrolysate in the intestine: Clathrin-dependent endocytosis as the main transport route.

This paper aimed to investigate the in vivo absorption of egg white hydrolysate (EWH) in rats and the transport route across the intestinal epithelium. Results showed that the level of plasma peptide-bound amino acid (PAA) of the EWH-supplemented rats (EWH-R) was determined to be 2012.18 ± 300.98 µmol/L, 10.72% higher than that of the control group, and was significantly positively correlated to that of EWH. Thirty-three egg white-derived peptides were successfully identified from the plasma of EWH-R, and 20 of them were found in both EWH-R plasma and EWH, indicating that these peptides tend to be absorbed through the intestinal epithelium in intact forms into the blood circulation. In addition, 637 up-regulated and 577 down-regulated genes in Caco-2 cells incubated with EWH were detected by RNA-sequencing and the clathrin-dependent endocytosis was the most enriched pathway in KEGG analysis. EWH significantly increased the mRNA levels of the key genes involved in the clathrin-dependent endocytosis but these changes would be inhibited by the clathrin-dependent endocytosis inhibitor of chlorpromazine. Moreover, the transepithelial transport of EWH across Caco-2 cell monolayers was significantly reduced by chlorpromazine. This study provided molecular-level evidence for the first time that clathrin-dependent endocytosis might be the main transport route of EWH in the intestinal epithelium.

Translucency mechanism of heat-induced pigeon egg white gel.

In this study, the properties of pigeon egg white (PEW) and chicken egg white (CEW) thermal gels were compared, with the aim of revealing the mechanisms involved in the high transparency of PEW thermal gels. Results demonstrated that PEW gels exhibited higher transparency than CEW gels. Scanning electron microscope (SEM) and transmission electron microscope (TEM) analysis revealed that PEW gels formed a fine chain gel network structure with an average diameter of thermal aggregates (89.84 ± 7.13 nm). The molecular properties of PEW proteins, such as higher content of ß-sheet structures (32.73 %), reactive groups (free sulfhydryl groups, hydrophobic groups), and absolute zeta potential (-3.563 mV), were found to contribute to the formation of smaller thermal aggregates during thermal denaturation. The microrheology measurements showed that these features allowed PEW proteins to interact less with each other and form smaller thermal aggregates during thermal denaturation, which facilitated the formation of fine chain gel networks and thus improved the transparency of the gels. The present study initially reveals the molecular basis of the high transparency of PEW thermal gels and provides a theoretical reference for the development of new highly transparent protein materials.

Fabrication mechanism and functional properties of ovalbumin fibrils prepared by acidic heat treatment.

BACKGROUND: Ovalbumin (OVA), accounting for 50% of proteins in egg white, is a kind of high-quality protein with excellent nutritional and processing functions. Acid heat treatment will induce the deformation and filtration of OVA, endowing it with improved functionality. However, the molecular kinetic process during the fibrillation of OVA and the application of the fabricated OVA fibrils (OVAFs) have not been thoroughly studied and revealed. RESULTS: In this study, the fabrication mechanism and the application OVAFs as an interfacial stabilizer and polyphenol protector were investigated. Acidic (pH 3.0) heat treatment was used to induce the fibrillation of OVA, and thioflavin T fluorescence intensity, molecular weight distribution, and the tertiary and secondary structures of OVAF samples were recorded to determine the fibrillation efficiency and the molecular mechanism. The results showed that, in the initial stage of fibrillation, OVA first hydrolyzed to oligopeptides, accompanied by the exposure of hydrophobic domains. Then, oligopeptides were connected by disulfide bonds to form primary fibril monomers. Hydrophobic interaction and hydrogen bonding may participate in the further polymerization of the fibrils. The fabricated OVAFs were characterized by a ß-sheet-rich structure and possessed improved emulsifying, foaming, and polyphenol protection ability. CONCLUSION: The research work was meaningful for exploring the application of globular water-soluble OVA in an emerging nutritious food with novel texture and sensory properties. © 2023 Society of Chemical Industry.

Preparation of egg white powder using electrohydrodynamic drying method and its effect on quality characteristics and functional properties.

This research investigated the effects of spray drying (SD, set at 180 °C), freeze-drying (FD, set at -35 °C), and electrohydrodynamic drying (EHD) with and without the foam-mat method on egg white. The configuration used in EHD was a wire-to-plate type at room temperature. The results showed no significant difference in gel hardness and WHC% (P ≥ 0.05). Also, the foam-mat EHD powders resembled the FD powders in microstructure, appearance, flowability, and absorption intensity of the Amide I and II bands. Furthermore, the foam-mat EHD (DC-) powder had the highest protein content (66.1%), enthalpy (-183.06 J/g), and foaming capacity (725%) (P < 0.05). This finding was proved by FTIR, Raman, and SDS-PAGE tests, which revealed the minor structural changes in proteins (peptide chain structure, Amide I, Amide II, α-helix, and ß-sheet). FD powder demonstrated good protein stability in zeta potential and foam stability tests.

Thermal gelation and digestion properties of hen egg white: Study on the effect of neutral and alkaline salts addition.

In this study, the thermal gelation and digestion properties of hen egg white (hen EW) proteins with different salts were investigated. Results show that the addition of neutral salt - sodium chloride (NaCl) decreased the gel hardness/resilience, increased gel lightness, aggregated particle size and digestibility of hen EW proteins significantly. In contrast, alkaline salts - phosphate and carbonate addition increased the gel resilience and strain tolerance as well as reduced the aggregated particle size and gel lightness of hen EW proteins due to the increase of solution pH and negative charge. Correlation analysis shows that the digestibility of hen EW gels was affected by gel viscoelasticity, molecule forces and texture. In conclusion, thermal gelation properties of hen EW proteins could be modulated by salts with different pH/ionic strength, and thus affected the protein digestion and peptide released.

Changes in partial properties of glycosylated egg white powder during storage.

BACKGROUND: Glycosylation is an effective method to modify protein. However, there is a lack of research on the property changes of glycosylated protein during storage. In the present study, the changes in the physicochemical, functional, and structural properties of xylo-oligosaccharide (XOS) glycosylated egg white powder (EWP) (XOS-EWP conjugates) prepared with different glycosylation conditions (XOS/EWP ratio and reaction time) were investigated when stored at 25 °C and 60% relative humidity. RESULTS: In the 12 weeks of storage, the degree of grafting, browning, and the formation of Maillard reaction products of XOS-EWP conjugates increased. The increase in XOS/EWP ratio and reaction time led to an increase in protein aggregation, though a decrease in solubility, due to increased degree of glycosylation and structural changes. Furthermore, improved gel hardness of XOS-EWP conjugates deteriorated, while improved emulsification ability was kept stable during storage. For the sample with a lower XOS/EWP ratio and reaction time, the gel hardness and emulsifying properties underwent little or no deterioration even improving during storage. The results could be attributed to the limited degree of glycosylation, further unfolding of the protein structure, increased surface hydrophobicity of protein, and improved thermal characteristics. CONCLUSION: During storage, the Maillard reaction would continue to occur in the glycosylated EWP, further affecting the performance of modified EWP. Modified EWP prepared under different glycosylation conditions performed differently during storage. Modified EWP with a larger XOS/EWP ratio and reaction time meant it was harder to maintain good performance. Modified EWP with a smaller XOS/EWP ratio and reaction time changed significantly to better performances. © 2022 Society of Chemical Industry.

Storage impact on egg white powder's physical and functional properties.

BACKGROUND: Changes in storage temperature and time alter the functional properties of egg white powder (EWP) and determine its quality and shelf-life, finally affecting the consumer acceptance of the products made from EWP. In the present study, the EWP samples were stored at four different temperatures (-20, 4, 25 and 37 °C) for 60 days, and then the protein structural, physical and functional properties of EWP were measured and assessed further for correlation with storage conditions using heatmap. RESULTS: The viscosity of the EWP solution increased after 30 days. Foaming ability and rheological properties increased first and then decreased compared to untreated samples with the prolonged storage time. Correlation analysis results indicated that the gel hardness, water holding capacity, foaming ability, emulsifying ability, particle size, dispersibility and viscosity of EWP were significantly related to storage time (P < 0.05). Only the gelation properties of EWP stored at 37 °C for 60 days changed significantly and were negatively related to its moisture content (P < 0.05). Additionally, the random coil content of EWP was positively correlated with particle size, moisture content, solubility and gel properties, whereas ß-sheet was negatively correlated with them. CONCLUSION: Compared to other temperatures, the functional properties of EWP were relatively stable under 4 °C. Therefore, the low temperature (4 °C) was selected as the most suitable storage temperature for EWP. The results of the present study could provide a theoretical basis for the shelf-life extension of EWP. © 2022 Society of Chemical Industry.

Recent development of egg protein fractions and individual proteins as encapsulant materials for delivery of bioactives.

Egg proteins, as one of the most abundant animal protein sources, have received considerable attention for developing delivery systems. Among all egg proteins, egg white (ovalbumin) is the most promising encapsulant due to its excellent properties such as gelling, digestibility, self-assembly, amphiphilic nature. In this review paper, we focused particularly on egg protein-based delivery systems with superior encapsulation and delivery functions, including polymeric nanoparticles, emulsions, hydrogels and aerogels. Egg protein-based delivery systems across a wide range of geometry and dimensions have been applied to protect or control-release bioactive small molecules and macromolecules, probiotics and metal nanostructures. However, there are challenges that must be carefully addressed for advancing the practical applications of egg protein-based delivery system in foods, including allergenicity from ovalbumin and ovotransferrin, intolerance to environmental conditions, limited processing technologies. More efforts are warranted to fill knowledge gaps related to fabrication, utilization and digestive mechanisms of egg protein-derived delivery systems.

Reactive Green 19 dye-ligand immobilized on the aminated nanofiber membranes for efficient adsorption of lysozyme: Process development and optimization in batch and flow systems.

The performance of lysozyme adsorption by the aminated nanofiber membrane immobilized with Reactive Green 19 (RG19) dyes was evaluated in batch and flow systems. The physicochemical properties of the dye-immobilized nanofiber membrane were characterized. The parameters of batch-mode adsorption of lysozyme (e.g., pH, initial dye concentration, and lysozyme concentration) were optimized using the Taguchi method. In a flow process, the factors influencing the dynamic binding performance for lysozyme adsorption in the chicken egg white (CEW) solution include immobilized dye concentration, adsorption pH value, feed flow rate, and feed CEW concentration. The impact of these operating conditions on the lysozyme purification process was investigated. Under optimal conditions, the recovery yield and purification factor of lysozyme achieved from the one-step adsorption process were 98.52% and 143 folds, respectively. The dye-affinity nanofiber membrane also did not exhibit any significant loss in its binding capacity and purification performance after five consecutive uses.

Long-term preservation at low temperature of Escherichia coli cells embedded in egg white glass formed by slow drying at room temperature.

Sterile homogeneous egg white (EW) is obtained through a three-step process, high-speed homogenization, centrifugation, and ultraviolet radiation. After incorporating 1.056 × 1010 CFU/g of Escherichia coli, the EW mixture was dehydrated by slow drying to form a brittle, water-soluble, and transparent bacteria-embedded egg white glass (BE-EWG). The BE-EWG stored at -20 °C for 4 months maintains almost all the cell growth functions and proliferation activities of the labeled E. coli, and most of the cell functions and 60 % of the proliferation activities are maintained for up to one year. The BE-EWG exhibits a porous hydrogel membrane structure after heat treatment, and many E. coli cells are accommodated in a grid with a pore size of 2-10 mm. The loss of bacteria-carrying viability after storage at room temperature may be related to the Maillard reaction between protein and glucose in EW, which results in the structural changes caused by protein cross-linking, darkened color and water insolubility of the BE-EWG. Therefore, the method of embedding E. coli cells in EWG as solid form at room temperature to avoid ice crystal formation during cryopreservation is more beneficial for storage, packaging and shipping at -20 °C.

Effects of different copper salts on the physicochemical properties, microstructure and intermolecular interactions of preserved egg white.

This study was conducted to investigate the effects of different copper salts on the physicochemical properties, microstructure and intermolecular interactions of preserved egg white (PEW) during pickling. With increased pickling time, the alkalinity of the pickling solution and the moisture of PEW pickled by three copper salts (CuSO4, CuCl2 and Cu(CH3COO)2) gradually decreased, while the pH showed dynamic trends. During the early stage of pickling, the viscosity of PEW changed dynamically. With the progress of pickling, the hardness and springiness of PEW increased gradually and the α-helix structure gradually transformed into a more stable ß-structure. The main forces that maintained PEW gel were ionic and disulfide bonds. In the early stage of pickling, the alkali penetration rate of CuCl2 pickled preserved eggs was faster, and better gel strength and network structure were observed, followed by CuSO4 and Cu(CH3COO)2, while there were no significant differences in the later stage of pickling.

Effects of κ-carrageenan on gel quality of threadfin bream (Nemipterus spp.) surimi containing salted duck egg white powder.

This study aimed to evaluate the combined effect of κ-carrageenan and salted duck egg white powder (SDEWP) in improving the gel quality of threadfin bream surimi. Effects of κ-carrageenan at different levels (0-2 %) on gel properties of threadfin bream surimi without and with salted duck egg white powder at 3 % (protein equivalent) were investigated. A combination of 0.5 % κ-carrageenan and SDEWP increased breaking force of surimi gel by 139.7 % and deformation by 55.1 %, compared to the control (P < 0.05). The expressible moisture content (EMC) was decreased by 50.0 % in the surimi gel added with 0.5 % κ-carrageenan and SDEWP. Hardness, cohesiveness, gumminess, and chewiness of surimi gel were also improved (P < 0.05). However, springiness of surimi gel was not affected. SDEWP reduced proteolytic degradation in surimi gel. Surimi gel with augmented whiteness was attained when κ-carrageenan was added at higher levels. Microstructure of surimi gel shown that the gel became denser and more uniform when added with 0.5 % κ-carrageenan and SDEWP. Therefore, κ-carrageenan can be used to enhance the effectiveness of SDEWP and further improve the gel quality of threadfin bream surimi added with SDEWP.

Enzymatic hydrolysis re-endows desalted duck egg white nanogel with outstanding foaming properties.

Heat-induced gel-assisted desalination could efficiently and inexpensively remove salt from salted egg whites. However, it was at the expense of the excellent foaming properties of egg whites, caused by the denaturation and aggregation of proteins during heating treatment. Hence, in this current work, the enzymatic treatment was used to re-endow duck egg white nanogels (DEWN) with outstanding foaming properties. We found that low levels of hydrolysis (DH = 2.27 %) could dramatically improve the foaming capability (FC), reaching >200 %, which also enhanced the foaming stability (FS). As the hydrolysis time extended, the adsorption and diffusion rate of the supernatant on the interface increased and performed high elasticity. The dilatational rheology and Lissajous plots were explored to investigate the nonlinear dilatational rheological behaviors of the air/water interface stabilized by the hydrolysed samples. Finally, we evaluated the effect of pH on foaming properties and found that the FC could exceed 250 %, and the FS was close to 80 % at pH = 5. These encouraging results showed that simple enzymatic treatment could revive nanogels from their dissatisfied foaming properties. In this work, gel-assisted desalination combined with enzyme treatment significantly promotes the high-quality and high-value utilization of salted egg white.

Improving the gel properties of duck egg white by synergetic phosphorylation/ultrasound: Gel properties, crystalline structures, and protein structure.

To improve the gel properties of duck egg white gel and increase the industrial value of duck egg white, the mechanisms of ultrasound and synergetic phosphorylation/ultrasound treatments were examined in this study. It was found that as the ultrasound power increased, the surface hydrophobicity, hardness, and cohesiveness of the gel system increased, and the ζ-potential and water mobility decreased. Of the two treatments, phosphorylation/ultrasound had the strongest impact on the conformation and crystallinity of the gel system and promoted the formation of high molecular polymers. Both gel systems displayed enhanced compactness, stability, and gel strength because of the enhanced protein-protein interactions via hydrogen bonds and protein aggregation, and increased the content of intramolecular ß-sheets following ultrasound treatment, and synergetic phosphorylation/ultrasound further improved the stability, water binding and gel properties. This experiment showed that ultrasound and, particularly, phosphorylation/ultrasound are effective methods to improve the gel properties of duck egg white. This study enhanced our understanding of the interactions of sodium pyrophosphate and egg white under ultrasound treatment, and promote the potential application of sodium pyrophosphate and ultrasound treatment of novel food products.

Role of salts and solvents on the defibrillation of food dye "sunset yellow" induced hen egg white lysozyme amyloid fibrils.

Several food dyes are known to induce amyloid fibrillation when interacting with proteins. Here, we studied the role of sunset yellow (SY) in the amyloid fibrillation of hen egg white lysozyme (HEWL) and characterized the changes using spectroscopy techniques. Turbidity results showed that SY dye induces aggregation in HEWL in concentrations dependent manner. The aggregation induced by SY dye is kinetically very fast, no lag phase was detected, and the kinetics process follows an isodesmic kinetics pathway. The SY-dye induce aggregates have cross-ß secondary structure confirmed by far-UV CD measurements. The effect of salts and solvents was also seen on SY-induced aggregates. Turbidity, far-UV CD, and kinetics results suggest that certain concentrations of NaCl and (NH4)2SO4 solubilize the SY-induce amyloid fibrils, but (NH4)2SO4 is more effective. Similarly, solvents are also solubilized the SY-induces HEWL amyloid fibrillation but the order of defibrillation is as follows: Isopropanol> ethanol > methanol which signified that isopropanol is more effective than other solvents. The salts and solvents data suggest that the electrostatic, as well as hydrophobic interaction, is responsible for SY-induced amyloid fibrillation. These conformational changes should be examined, more seriously for the purpose of food safety.

Identification and Immunomodulatory Effect on Immunosuppressed Mice of Selenium-Enriched Peptides of Egg White.

Selenium-enriched egg white peptides (Se-EWP) were prepared by pre-heat treatment and enzymatic hydrolysis in this study. In addition, their selenopeptide sequence identification and immunomodulatory effect were investigated. Results showed that the yield of Se-EWP obtained from alkaline-neutral protease treatment reached 76.90%, and peptides with a molecular weight of 200-1000 Da accounted for 98.33%. Four characteristic selenopeptides, including SeCys-Trp-Leu-Glu, Trp-Ser-SeCys, SeMet-Ala-Pro, and SeMet-Leu, were identified by HPLC-ESI-MS/MS, which were rich in hydrophobic and branched-chain amino acids. Se-EWP (750 mg/kg/d) could effectively retard the decrease of immune organ index in immunosuppressed mice induced by cyclophosphamide. Moreover, supplementation of Se-EWP could promote a higher content of Se in liver, the number of white blood cells, and the levels of serum cytokines (IL-6, IL-2, and TNF-α) as compared with EWP groups, indicating that Se-EWP could effectively alleviate immunosuppression induced by cyclophosphamide. These findings suggested that Se-EWP exhibited great potential as functional foods for immunomodulatory effect.

Formation mechanism of high-viscosity gelatinous egg white among "Fenghuang Egg": Phenomenon, structure, and substance composition.

"Fenghuang Egg" is a special egg product incubated for 12 days by fertilized hen eggs. Its egg white contains high-viscosity and excellent thermal gel strength. A comparative study on the differences in gel properties, structure, and substance composition between fresh egg white (FEW) and "Fenghuang egg" gelatinous egg white (GEW) was carried out. Experimental results showed GEW had better apparent viscosity, as well as the hardness, cohesiveness and water holding capacity (WHC) of thermal gel; the content and size of aggregate structure increased significantly in GEW, and a fibrous dense network composed of numerous spherical nanoparticles connected in series was formed after heating. In addition, it also discovered that more water molecules in GEW existed in the form of bound water. A total of 41 proteins changed significantly in FEW and GEW, Mucin 6 might be the main reason for the enhanced viscosity of GEW, and OVA might be the dominant protein differentiating the thermal gel properties between FEW and GEW. This study revealed that the differences in gel properties and structures between FEW and GEW were closely related to the content of highly glycosylated globular proteins, laying a theoretical foundation for the application of high-viscosity egg whites.