Chicken Egg White Gels: Fabrication, Modification, and Applications in Foods and Oral Nutraceutical Delivery.

Chicken egg white (EW) proteins possess various useful techno-functionalities, including foaming, gelling or coagulating, and emulsifying. The gelling property is one of the most important functionalities of EW proteins, affecting their versatile applications in the food and pharmaceutical industries. However, it is challenging to develop high-quality gelled foods and innovative nutraceutical supplements using native EW and its proteins. This review describes the gelling properties of EW proteins. It discusses the development and action mechanism of the physical, chemical, and biological methods and exogenous substances used in the modification of EW gels. Two main applications of EW gels, i.e., gelling agents in foods and gel-type carriers for nutraceutical delivery, are systematically summarized and discussed. In addition, the research and technological gaps between modified EW gels and their applications are highlighted. By reviewing the new modification strategies and application trends of EW gels, this paper provides insights into the development of EW gel-derived products with new and functional features.

Cystatins: unravelling the biological implications for neuroprotection.

Cystatins, a family of proteins known for their inhibitory role against cysteine proteases, have garnered significant attention in the field of neurodegeneration. Numerous genetic, experimental, and clinical studies concerning cystatin C suggest it plays an important role in the course of neurodegenerative diseases. Its beneficial effects are associated with cysteine protease inhibition, impact on ß-amyloid aggregation, as well as regulation of cell proliferation, autophagy, and apoptosis. Cystatin isolated from chicken egg white, called ovocystatin, has been widely used in medical and pharmaceutical research due to its structural and biological similarities to human cystatin C. This article focuses on the potential use of cystatins, with special emphasis on easily obtained ovocystatin, in the treatment of neurodegenerative diseases, such as dementia. The current evidence on cystatin use has shed light on its mechanisms of action and therapeutic implications for neuroprotection and maintenance of cognitive functions.

UiO66-based molecularly imprinted polymers with water-compatible deep eutectic solvent as functional monomer for purification of lysozyme from egg white.

Protein-templated molecularly imprinted polymers have limitations such as poor mass transfer, slow recognition kinetics, and difficulties in isolation and purification due to their large molecular sizes, complex structures, and flexible conformations. To address these limitations and obtain lysozyme (Lyz)-imprinted polymers, a molecularly imprinted polymer (UiO66@DES-MIPs) was prepared for the first time by using Lyz as a template molecule, a metal-organic framework (UiO66-NH2) as a matrix, and a water-compatible deep eutectic solvent (DES) as a functional monomer. The introduction of UiO66-NH2 by the solvothermal method with a large specific surface area and favorable stability and resistance to environmental disturbances into the MIPs can reduce the "embedding" phenomenon and acquire a higher binding capacity and fast mass transfer. In addition, a water-soluble binary DES (1:2 molar ratio of choline chloride to 1,3 dimethylurea) prepared by a hydrothermal method as a functional monomer generates multiple forces with Lyz, increasing the hydrophilicity of UiO66@DES-MIPs and contributing to the formation and stabilization of the imprinted sites. Consequently, UiO66@DES-MIPs exhibited good selectivity, water compatibility, and fast adsorption equilibrium (the adsorption equilibrated at 243.87 ± 4.88 mg g-1 in 90 min). Besides, reusability experiments indicated that the UiO66@DES-MIPs could be recycled six times without obvious loss of adsorption capacity. The imprinting factor of UiO66@DES-MIPs is 3.67. The isolation and purification of Lyz from egg white confirmed the practicability of UiO66@DES-MIPs. The high adsorption capacity and specific recognition make this polymer a promising candidate for the isolation and purification of biological macromolecules.

Functional capacity and quality of life improvement in stable chronic obstructive pulmonary disease (COPD) patients following physical exercise and chicken egg white supplementation.

The pillars of comprehensive pulmonary rehabilitation program for chronic obstructive pulmonary disease (COPD) patients include physical exercise and good nutrition. The aim of this study was to evaluate the efficacy of pulmonary rehabilitation, which included physical exercise and chicken egg white supplementation, on the quality of life (QoL) and functional capacity among patients with stable COPD. The COPD patients were enrolled prospectively in this quasi-experimental study and completed a 12-week smartphone-guided home-based physical exercise program that comprised strength and resistance training three times per week for 30 minutes each session. Participants were divided into two groups: the control group who underwent physical exercise only, and the intervention group who had physical exercise and chicken egg white supplementation as a protein source. Patient characteristics including sex, age, nutritional status, comorbidities, smoking status, and obstruction severity, were evaluated. The COPD assessment test (CAT) score and six-minute walk test (6MWT) were used as the parameters to evaluate QoL and functional capacity, respectively. Of the total 50 patients included in the study, 12 were excluded due to follow-up and adherence problems. Our data indicated there were significant CAT score reduction and 6MWT improvement in both control and intervention groups after 12 weeks compared to baseline data. However, reduction of mean CAT score was higher in intervention compared to control group (-13.47±6.49 vs -5.42±5.07, p<0.001). In addition, the improvement of 6MWT was also higher in intervention group compared to control group (145.47±69.2 vs 32.42±17.3 meters, p<0.001). In conclusion, chicken egg white supplement to male patients with stable COPD who exercise with resistance and strength training could improve the QoL and functional capacity.

Functional Properties and Extraction Techniques of Chicken Egg White Proteins.

Chicken egg whites contain hundreds of proteins, and are widely used in the food, biological and pharmaceutical industries. It is highly significant to study the separation and purification of egg white proteins. This review first describes the structures and functional properties of several major active proteins in egg whites, including ovalbumin, ovotransferrin, ovomucoid, lysozyme, ovomucin, ovomacroglobulin and avidin. Then, the common techniques (including precipitation, chromatography and membrane separation) and some novel approaches (including electrophoresis, membrane chromatography, aqueous two-phase system and molecular imprinting technology) for the separation and purification of egg white proteins broadly reported in the current research are introduced. In addition, several co-purification methods for simultaneous separation of multiple proteins from egg whites have been developed to improve raw material utilization and reduce costs. In this paper, the reported techniques in the last decade for the separation and purification of chicken egg white proteins are reviewed, discussed and prospected, aiming to provide a reference for further research on egg proteins in the future.

Selective isolation and sensitive detection of lysozyme using aptamer based magnetic adsorbent and a new quartz crystal microbalance system.

Magnetic chitosan beads and quartz crystal microbalance chip were decorated with lysozyme specific aptamer for isolation and detection of lysozyme, respectively. The lysozyme specific aptamer was immobilized on poly (dopamine) coated magnetic chitosan beads and the chip via Schiff base reaction. The percentage of the removal efficiency and purity of the isolated lysozyme from egg white were 87.6% and 91.8%, respectively. Further, the sensor system was contacted with different concentrations of lysozyme and other test proteins. This sensor system provided a method for the label-free, concentration-dependent, and selective detection of lysozyme with an observed detection limit of 17.9 ± 0.6 ng/mL. The sensor system was very selective and not significantly responded to the other tested proteins such as ovalbumin, trypsin, cytochrome C, and glucose oxidase. The prepared new sensor system showed a good durability and a high sensitivity for determination of lysozyme from solutions and whole egg white.

One‒pot green synthesized protein‒based silver nanocluster as prooxidant biosensor.

In this study, silver nanoclusters as prooxidant biosensor were eco‒friendly synthesized using chicken egg white protein without any chemical reducing agents for measuring copper(II)-induced prooxidant activities of catechin, epicatechin, epigallocatechin gallate, resveratrol, gallic acid, chlorogenic acid, and rutin. The prooxidant activities were evaluated via measuring the absorption at 450 nm wavelength of the Cu(I)‒neocuproine chelate formed by extraction of protein-bound Cu(I) with neocuproine reagent. Accuracy was determined by evaluating recovery values of wine, grape and apple samples and the obtained values were between 97.2%‒98.9%. Intra-day precision and inter-day reproducibility experiments were studied with three different experiments in a day and three different days respectively. The obtained relative standard deviation values were 0.96% and 1.91%. The detection limit of the biosensor was found as 0.2 µM. The total prooxidant activities of fresh apple and grape fruits, apple and grape juices, and red wine were determined and the results obtained were compared with the findings of the carbonyl assay. In this study, a cheap, easily applicable, sensitive, and reproducible biosensor was developed. It was seen that it could be used in the measurement of the prooxidant activity of different food samples and give an idea about diet, healthy life, and nutrition.

Biocatalyst and colorimetric biosensor of carcinoembryonic antigen constructed via chicken egg white-copper phosphate organic/inorganic hybrid nanoflowers.

In this article, the dual-functional chicken egg white-copper phosphate organic-inorganic hybrid nanoflowers (Cu-NFs), combining the functions of signal amplification and biological recognition, were prepared through a simple one-pot method. The Cu-NFs exhibit excellent biocatalytic activity of peroxidase and polyphenol oxidase. Besides, a biotin-labeled secondary antibody encapsulated Cu-NFs-2 (Cu-NFs-2@Biotin-NHS-Ab2) capture probe was prepared by using the interaction between avidin in the egg white and biotin. Based upon this superiority, the as-prepared Cu-NFs-2 were used in labeled avidin-biotin enzyme-linked immunosorbent assay (Cu-NFs-2 based-LAB-ELISA) to construct a sensitive colorimetric biosensor for the ultrasensitive detection of carcinoembryonic antigen (CEA). Under weak alkaline (pH = 7.5) conditions, the as-developed colorimetric sensor displayed a wide linear range of 0.05-40 ng/mL with a detection limit of 3.52 pg/mL. Furthermore, this colorimetric sensor has been successfully applied to the detection of CEA in human serum samples. Therefore, the as-developed colorimetric sensor has broad application prospects in the field of medical diagnosis and portable detection.

A novel nonenzymatic ascorbic acid electrochemical sensor based on gold nanoparticals-chicken egg white-copper phosphate-graphene oxide hybrid nanoflowers.

Au-CEW-Cu3(PO4)2-GO nanoflowers (HNFs), which were assembled of gold nanoparticals (Au NPs), chicken egg white (CEW), copper phosphate (Cu3(PO4)2) and graphene oxide (GO) together to form a flower-like organic/inorganic hybrid nanocomposite, were synthesized through a simple and gentle one-pot co-precipitation method. The prepared samples were well characterized by scanning electron microscope, transmission electron microscope, energy dispersive x-ray spectrometer, x-ray diffraction and Raman spectrometer. The prepared Au-CEW-Cu3(PO4)2-GO HNFs was used to modify glassy carbon electrode to fabricate an electrochemical sensor for detection of ascorbic acid (AA). The electrochemical test results show that the linear range of the developed sensor is 8-300µM and the detection limit is 2.67µM (S/N = 3). While this sensor displays high sensitivity of 6.01 × 10-3µAµM-1cm-2and low detection potential of 35 mV due to the combination of the high conductivity of Au NPs, the larger specific surface area of GO and the intrinsic electrocatalytic activity of CEW-Cu3(PO4)2HNFs. Moreover, the Au-CEW-Cu3(PO4)2-GO HNFs-based sensor was successfully developed for application in electrochemical detection of AA in vitamin C tablets.

Adsorption and purification performance of lysozyme from chicken egg white using ion exchange nanofiber membrane modified by ethylene diamine and bromoacetic acid.

A high-performance polyacid ion exchange (IEX) nanofiber membrane was used in membrane chromatography for the recovery of lysozyme from chicken egg white (CEW). The polyacid IEX nanofiber membrane (P-BrA) was prepared by the functionalization of polyacrylonitrile (PAN) nanofiber membrane with ethylene diamine (EDA) and bromoacetic acid (BrA). The adsorption performance of P-BrA was evaluated under various operating conditions using Pall filter holder. The results showed that optimal conditions of IEX membrane chromatography for lysozyme adsorption were 10% (w/v) of CEW, pH 9 and 0.1 mL/min. The purification factor and yield of lysozyme were 402 and 91%, respectively. The adsorption process was further scaled up to a larger loading volume, and the purification performance was found to be consistent. Furthermore, the regeneration of IEX nanofiber membrane was achieved under mild conditions. The adsorption process was repeated for five times and the adsorption capacity of adsorber was found to be unaffected.

The effects of chicken egg white cystatin and proteinase inhibitor on cysteine peptidase-like activity in the sera of patients with breast cancer.

BACKGROUND: The activity of autogenic proteolytic enzymes is regulated in vivo by autogenic inhibitors. They play important roles in maintaining a balance in many processes in the human body. In pathological conditions, enzymes are overexpressed and the balance is disturbed. Such uncontrolled changes may lead to the development of local or systemic cancer. OBJECTIVES: To evaluate the effects of specific inhibitors, i.e., chicken egg white cystatin (CEWC) and proteinase inhibitor (E-64) on autogenic cysteine peptidases (CPs) in the sera of patients reporting for subsequent stages of treatment after being diagnosed with breast cancer. Cysteine peptidases play a vital role in the basic processes that are associated with cancer progression. MATERIAL AND METHODS: We selected serum samples from 108 patients with a diagnosis of breast cancer (stages IIA-IIIA) who had received no previous treatment. The blood samples were centrifuged, and the resulting serum was placed in liquid nitrogen and stored at -80°C. The biochemical tests were performed at the laboratory of the Department of Physical Chemistry and Microbiology. RESULTS: For CEWC, we found an inhibitory effect in 37 out of 108 samples; for E-64, 14 out of 22 samples displayed an inhibitory effect. In the remaining blood samples, these inhibitors caused an increase in fluorescence. In a parallel test, we added pure cathepsin B to 9 serum samples, and then used CEWC to inhibit the activity of autogenic CPs. Chicken egg white cystatin completely inhibited the cathepsin B that was added to the serum without changing its effect on the autogenic CPs. CONCLUSIONS: The results suggest that there may be a potential difference between the commercially available cathepsin B and its autogenic analogues found in the serum of cancer patients. The increase in fluorescence induced in the reaction between the inhibitors and autogenic CPs is still unexplained. There was no relationship between the observed inhibition/activation of CPs and any of the available indicators of the health of the patients examined.

Use of magnetic Fe3O4 nanoparticles coated with bovine serum albumin for the separation of lysozyme from chicken egg white.

Fe3O4 magnetic nanoparticles modified with tetraethyl orthosilicate and bovine serum albumin (Fe3O4@TEOS@BSA) were synthesized and efficiently used to separate lysozyme from egg white. Glutaraldehyde was used to crosslink the bovine serum albumine molecules around the nanoparticles. The surface modifications were attested by transmission electron microscopy, infrared spectroscopy, thermogravimetry analysis, and zeta potential. The material was thermally stable, and its surface charge was pH-dependent. The best lysozyme adsorption and desorption were obtained at pHs 10.0 and 5.0, respectively. The pseudo-second-order model fitted well into the lysozyme adsorption kinetic data and the time for the equilibrium was 15 min. The adsorption equilibrium results were best described by the Freundlich model. Fe3O4@TEOS@BSA particles were very efficient to extract lysozyme from chicken egg, according to the SDS-PAGE analyses. The extracted molecules maintained their enzymatic activity in about 90%. Fe3O4@TEOS@BSA particles were easily recycled, with their reuse being possible 5 times with the same performance.

Different fluorescence emitting copper nanoclusters protected by egg white and double-emission fluorescent probe for fast detection of ethanol.

Green emitting copper nanoclusters (G-Cu NCs), yellow emitting Cu NCs (Y-Cu NCs), orange emitting Cu NCs (O-Cu NCs) and red emitting Cu NCs (R-Cu NCs) were prepared using chicken egg white as the stabilizer by changing the reaction conditions. This is a green, facile and cheap method to explore different color emitting CuNCs by the same precursor and stabilizers. The G-Cu NCs were employed for the detection of ethanol due to their aggregation induced emission enhancement (AIEE) effect. The fluorescence emission of Cu NCs at 526 nm under the excitation of 444 nm can be effectively enhanced in the presence of ethanol due to AIEE effect, thus realizing the quantitative determination of ethanol content in the range 5-60%. In addition, a visual dual-emission fluorescence probe with the combination of G-Cu NCs and silicon nanoparticles (Si NPs/G-Cu NCs) was designed to evaluate ethanol content conveniently and rapidly. Desirable linear relationship is observed between ratio of fluorescence intensity (I525/I441) and ethanol content under the excitation of 383 nm. Visible color transformation of this probe is observed in the ethanol content range 2-20%. Moreover, the ethanol sensing platforms were applied to the detection and evaluation of the alcohol content of liquor, and the recoveries in liquor were in the range 99.7% to 113%, broadening the applications of Cu NCs and providing a sensitive detection method for ethanol.

Purification of lysozyme from chicken egg white by high-density cation exchange adsorbents in stirred fluidized bed adsorption system.

Lysozyme from crude chicken egg white (CEW) feedstock was successfully purified using a stirred fluidized bed adsorption system ion exchange chromatography where STREAMLINE SP and SP-XL high density adsorbents were selected as the adsorption carrier. The thermodynamic and kinetic studies were carried out to understand the characteristics of lysozyme adsorption by adsorbents under various conditions, including adsorption pH, temperature, lysozyme concentration and salt concentrations. Results showed that SP and SP-XL adsorbents achieved optimum lysozyme adsorption at pH 9 with capacity of ~139.77 and ~251.26 mg/mL, respectively. The optimal conditions obtained from batch studies were directly employed to operate in SFBA process. For SP-XL adsorbent, the recovery yield and purification factor of lysozyme were 93.78% and ~40 folds, respectively. For SP adsorbent, lysozyme can be eluted ~100% with purification factor of ~26 folds. These two adsorbents are highly suitable for use in direct recovery of lysozyme from crude CEW.

A Comprehensive Identification of Chicken Egg White Phosphoproteomics Based on a Novel Digestion Approach.

There are plenty of phosphoproteins in chicken egg white (CEW), which are of great significance for the biological activity and function of CEW. In this study, phosphorylated proteins in CEW were identified and analyzed based on two digestion strategies (trypsin and trypsin/glutamyl endoproteinase). Besides, the enrichment strategy of immobilized metal affinity chromatography was used, and phosphopeptides were identified by nano liquid chromatography/tandem mass spectrometry. A total of 189 phosphosites mapped onto 166 phosphopeptides corresponding to 96 phosphoproteins were identified. Gene ontology analysis suggested that these phosphoproteins of CEW mainly participated in biological processes such as "cell process", "biological regulation", and "response to stimulus". Moreover, the phosphoproteins of CEW were involved in molecular functions, primarily including "binding" and "catalytic activity". On the basis of the available literature, the research was the first comprehensive identification of chicken egg white phosphoproteins. This study further enriched the identification of phosphoproteins in CEW and laid a foundation for the subsequent study of phosphoproteins.

Effective purification of lysozyme from chicken egg white by tris(hydroxymethyl)aminomethane affinity nanofiber membrane.

Polyacrylonitrile nanofiber membrane functionalized with tris(hydroxymethyl)aminomethane (P-Tris) was used in affinity membrane chromatography for lysozyme adsorption. The effects of pH and protein concentration on lysozyme adsorption were investigated. Based on Langmuir model, the adsorption capacity of P-Tris nanofiber membrane was estimated to be 345.83 mg/g. For the operation of dynamic membrane chromatography with three-layer P-Tris nanofiber membranes, the optimal operating conditions were at pH 9, 1.0 mL/min of feed flow rate, and 2 mg/mL of feed concentration. Chicken egg white (CEW) was applied as the crude feedstock of lysozyme in the optimized dynamic membrane chromatography. The percent recovery and purification factor of lysozyme obtained from the chromatography were 93.28% and 103.98 folds, respectively. Our findings demonstrated the effectiveness of P-Tris affinity nanofiber membrane for the recovery of lysozyme from complex CEW solution.

Removal of dye waste by weak cation-exchange nanofiber membrane immobilized with waste egg white proteins.

In this research, a protein nanofiber membrane (P-COOH-CEW) was developed to treat the dye waste. Initially, polyacrylonitrile nanofiber membrane (PAN) was prepared by electrospinning, followed by heat treatment, alkaline treatment, and neutralization to obtain weak cation exchange nanofiber membrane (P-COOH). The P-COOH membrane was chemically coated with chicken egg white (CEW) proteins to obtain a 3D structure of complex protein nanofiber membrane (P-COOH-CEW). The composite prepared was characterized with Fourier Transform Infrared analysis (FTIR), Scanning Electron Microscopy (SEM), and thermogravimetric analysis (TGA). Further, the composite was evaluated by investigating the removal of Toluidine Blue O (TBO) from aqueous solutions in batch conditions. Different operating parameters - coupling of CEW, shaking rate, initial pH, contact time, temperature, and dye concentration were studied. From the results, maximum removal capacity and equilibrium association constant was determined to be 546.24 mg/g and 10.18 mg/mg, respectively at pH 10 and 298 K. The experimental data were well fitted to pseudo-second order model. Furthermore, desorption studies revealed that the adsorbed TBO can be completely eluted by using 50% ethanol or 50% glycerol in 1 M NaCl solution. Additionally, the reuse of P-COOH-CEW membrane reported to have 97.32% of removal efficiency after five consecutive adsorption/desorption cycles.

A novel gold nanocluster-based fluorometric biosensor for measuring prooxidant activity with a large Stokes shift.

A chicken egg white protein-protected gold nanocluster (CEW-AuNC) based fluorogenic biosensor, where protein was used as both reducing and protecting agent, was developed to determine the Cu(II)-induced prooxidant activity of natural antioxidants abundant in food and biological samples. Gold nanoclusters, prepared using egg white proteins, exhibited strong fluorescence. The prooxidant activity of the tested antioxidants was indirectly measured by their reducing action on Cu(II) to Cu(I), and the reduced cuprous ion was bound to the thiol groups in the CEW-AuNC structure, causing a decrease in fluorescence intensity. Epicatechin, catechin, epigallocatechin gallate, morin, rutin, quercetin, gallic, chlorogenic, and rosmarinic acids, glutathione, cysteine, N-acetyl cysteine, bilirubin, resveratrol, and α-tocopherol were studied as natural antioxidants. A fluorometric method showing a large Stokes shift with excitation/emission maxima at 360∕640 nm was developed to sensitively measure the decrease in the fluorescence of CEW-AuNC associated with the binding of copper(I) to the protein structure. Total prooxidant activities of the binary, ternary, and quaternary synthetic mixtures and of some food and synthetic serum samples were determined. The biosensor response was statistically compared to that of its spectrophotometric counterpart. This method can be used for the control of the oxidative stability of foods with a prolonged shelf life.

Purification of lysozyme from chicken egg white using diatom frustules.

A nontoxic chromatographic matrix, with low cost and high adsorption capacity, is always a major goal for therapeutic protein purification. In this study, the frustules from two cultured diatoms, Nitzschia bilobata (AQ1) and Psammodictyon panduriforme (NP), were investigated as cation exchange materials for lysozyme purification from chicken egg white. The surface area and cation exchange capacity of frustules were about 400 m2/g and 140 µmol/mL for AQ1, 390 m2/g and 130 µmol/mL for NP. The optimal pH was 9 for adsorption. Through batch purification, the lysozyme recovery was 86% with a purity of 95% by AQ1 frustules, which was higher than that by NP frustules (82% with a purity of 90%). In the flow-through system, the purity using AQ1 frustules notably increased to 99%, higher than the result of 91% using NP frustules. Diatom frustules from AQ1 are more effective and could be an alternative chromatographic matrix for lysozyme purification.

Hydroxyl radical-induced early stage oxidation improves the foaming and emulsifying properties of ovalbumin.

As the most abundant protein in chicken eggs, ovalbumin plays an important role in the processing of high value-added poultry products. The present study investigated the effects of hydroxyl radical-induced early stage oxidation on the physicochemical and interfacial properties of chicken egg white ovalbumin. Protein carbonyl content of ovalbumin increased (from 0.78 to 1.13 nmol/mg) with the oxidation time (0-5 h), while free sulfhydryl content (from 0.43 to 0.09 nmol/mg) and free amino group content (from 0.49 to 0.43 nmol/mg) decreased. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed that the exposure of ovalbumin to hydroxyl radicals caused self-cross-linking and resulted in the formation of dimers and trimers. Accompanied by these changes, the surface hydrophobicity of ovalbumin was enhanced about 1.5-fold with the deepening of oxidation, and the value of zeta potential became more negative (from -7.15 to -20.51 mv). About 2 h of moderate oxidation improved the foaming and emulsifying properties of ovalbumin (1.2-fold to 1.8-fold), while excessive oxidation (3 h) decreased these interface properties. Hydroxyl radical-induced oxidation changed the surface chemical groups and structures of ovalbumin, thereby affecting the surface properties. The foaming and emulsifying properties of ovalbumin could be improved by oxidation, increasing the application possibilities of ovalbumin in the food interface system.