Major Egg White Proteins Research Articles

Quantitative proteomics provides new insights into the mechanism of improving rehydration of egg white powder by ultrasonic pretreatment

Poor rehydration is one of the key factors affecting the functional properties of egg white powder (EWP). Reducing rehydrated precipitates is important for the processing and application of EWP. In this study, effects of ultrasonic pretreatment on the physicochemical and functional properties of EWP rehydration solutions were studied with the aim of revealing the mechanism of ultrasonic pretreatment to improve rehydration. Compared with freeze-dried EWP (FD) and spray-dried EWP (SD), the percentage of ultrasonic pretreated FD (UFD) and ultrasonic pretreated SD (USD) rehydrated precipitates decreased by 13.0 % and 5.6 %, respectively, after ultrasonic pretreatment (0.25 W/mL for 10 min); and the average particle sizes of UFD and USD solutions decreased by 22.5 % and 15.5 %, respectively. Fourier transform infrared spectroscopy showed that ultrasonic pretreatment caused higher β-sheet content in the protein secondary structure of UFD rehydrated precipitates (49.2 %). Quantitative proteomic analysis revealed a decrease in the abundance of major egg white proteins (ovalbumin, ovotransferrin, ovomucoid and ovomucin) in the rehydrated precipitates of UFD, except for lysozyme. It was also shown that lysozyme-centered aggregates were disrupted in the rehydrated precipitates of UFD. Our research suggests that ultrasonic pretreatment improves EWP rehydration by reducing the interactions between high abundance proteins as well as improving the solubility of high abundance proteins.

Cross-Linking of White Proteins in Soft-Shell Turtle Eggs Using Transglutaminase, Reductant, and Thermal Pretreatment

The effects of transglutaminase (TGase), reductant, and thermal treatment on the cross-linking of white proteins in soft-shell turtle eggs were investigated. Egg white proteins were denatured by reductant (0.83% 2-mercaptoethanol, 2-ME) pretreatment and thermal pretreatment (95 °C and 5 min), and the denatured proteins were then catalyzed by TGase (1.0 unit/mL). SDS–PAGE showed that without any pretreatments, three major egg white proteins (210 kDa, 115 kDa, and 76 kDa proteins) were inferior substrates for TGase. Only portions of the 210 kDa protein (7.9%), 115 kDa protein (11.4%), and 76 kDa protein (42.9%) were polymerized by TGase into high-molecular-weight (MW) protein polymers (>180 kDa) after incubation for 3 h at 40 °C. However, the combined use of TGase with 0.83% 2-ME and thermal pretreatment led to a significant increase (p < 0.05) in the rate of white protein polymerization after 3 h: 210 kDa protein (90.8%), 115 kDa protein (69.5%), and 76 kDa protein (72.2%). Particle size analysis indicated that these cross-linked high-MW protein polymers were 2000–10,000 nm in size. Based on the experimental results, egg white proteins denatured by 2-ME and heat pretreatment are more prone to TGase-induced cross-linking.

Enzymatic Hydrolysis of Ovotransferrin and the Functional Properties of Its Hydrolysates.

Bioactive peptides have great potentials as nutraceutical and pharmaceutical agents that can improve human health. The objectives of this research were to produce functional peptides from ovotransferrin, a major egg white protein, using single enzyme treatments, and to analyze the properties of the hydrolysates produced. Lyophilized ovotransferrin was dissolved in distilled water at 20 mg/mL, treated with protease, elastase, papain, trypsin, or α-chymotrypsin at 1% (w/v) level of substrate, and incubated for 0–24 h at the optimal temperature of each enzyme (protease 55°C, papain 37°C, elastase 25°C, trypsin 37°C, α-chymotrypsin 37°C). The hydrolysates were tested for antioxidant, metal-chelating, and antimicrobial activities. Protease, papain, trypsin, and α-chymotrypsin hydrolyzed ovotransferrin relatively well after 3 h of incubation, but it took 24 h with elastase to reach a similar degree of hydrolysis. The hydrolysates obtained after 3 h of incubation with protease, papain, trypsin, α-chymotrypsin, and after 24 h with elastase were selected as the best products to analyze their functional properties. None of the hydrolysates exhibited antioxidant properties in the oil emulsion nor antimicrobial property at 20 mg/mL concentration. However, ovotransferrin with α-chymotrypsin and with elastase had higher Fe3+-chelating activities (1.06±0.88%, 1.25±0.24%) than the native ovotransferrin (0.46±0.60%). Overall, the results indicated that the single-enzyme treatments of ovotransferrin were not effective to produce peptides with antioxidant, antimicrobial, or Fe3+-chelating activity. Further research on the effects of enzyme combinations may be needed.

Functional properties of ovotransferrin from chicken egg white and its derived peptides: a review

With emerging trends in the food and pharmaceutical industries, potential applications of egg-derived bioactive compounds were recognized. Ovotransferrin is a major egg white functional protein responsible for multiple bioactivities. The objectives of this review are to provide scientific evidence of the functional properties of chicken ovotransferrin and its derived peptides and to identify future research approaches and applications. Various easy, economical, and non-toxic methods have been reported to produce ovotransferrin with high yield and purity, and chemical and enzymatic approaches have been employed to release bioactive peptides. The native ovotransferrin is known to have antimicrobial, antioxidant, anticancer, and immunomodulatory activities. The peptides produced from ovotransferrin also are reported to have antioxidant, antimicrobial, antihypertensive, and anticancer properties. However, little or no application of these compounds in the food and pharmaceutical areas is available yet. Therefore, the practical application of OTF in nutraceutical and pharmaceutical areas are among the emerging areas of research.

Production and characterization of eggs from hens with ovomucoid gene mutation

Ovomucoid is a major egg white protein which is considered as the most dominant allergen in chicken eggs. Owing to the difficulty of separating ovomucoid from egg whites, researchers have adopted genetic deletion for development of hypoallergenic eggs. Previously, we used CRISPR/Cas9 to establish chickens with ovomucoid gene (OVM) mutations, but it remained unknown whether such hens could produce eggs at maturity. Here, we have reported on eggs laid by OVM-targeted hens. Except for watery egg whites, the eggs had no evident abnormalities. Real-time PCR revealed alternative splicing of OVM mRNA in hens, but their expression was limited. Immunoblotting detected neither mature ovomucoid nor ovomucoid-truncated splicing variants in egg whites. Sixteen chicks hatched from 28 fertilized eggs laid by OVM-targeted hens, and fourteen of the sixteen chicks demonstrated healthy growth. Taken together, our results demonstrated that OVM knockout could almost completely eliminate ovomucoid from eggs, without abolishing fertility. Thus, the eggs developed in this study have potential as a hypoallergenic food source for most patients with egg allergies.

Mechanism study on enhanced foaming properties of individual albumen proteins by Lactobacillus fermentation

Our recent research showed that moderate Lactobacillus fermentation endowed egg white with stronger foaming properties. But the individual contribution of albumen proteins and the relative mechanisms during the process were still not clear. In this study, three major egg white proteins (ovalbumin, ovomucin and ovomucoid) were singly fermented by multiple Lactobacillus (Lactobacillus delbruekii ssp. bulgaricus, Lactobacillus acidophilus, Streptococcus thermophilus and Bifidobacterium animalis) for 0, 3, 6 and 9 h, and the surface activity, degradation, apparent viscosity and foaming properties were investigated. For ovalbumin, the surface hydrophobicity largely increased at 3 h and then began to decrease; combined with the gradual degradation by protease, the foaming ability of ovalbumin significantly increased at 3 h and then decreased at 6 and 9 h of fermentation. The fermented ovomucin and ovomucoid possessed an improved surface characteristics and appropriate apparent viscosity; correspondingly, the both proteins exhibited the enhanced foaming ability and stability after fermentation. This work revealed the molecular mechanism behind Lactobacillus fermentation treatment increased foaming properties of egg white, and provided basic information on the application of fermentation approach in improving functionalities of egg white in industry.

Isolation, Purification and Antimicrobial Efficacy of Ovotransferrin from Eggs of Broiler, Domestic and Quail

Ovotransferrin is one of the major egg white proteins that have antimicrobial activity as well as iron binding capability. The objective of this study was to develop a simple and easy method to separate ovotransferrin using organic solvents. The purity of the ovotransferrin was determined using SDS-PAGE, the isolated protein was further studied for the assessment of interaction between food borne pathogens and the isolated Otrf by protein-protein molecular docking. However, this method is simpler and more cost effective than the previous method. The isolated ovotransferrin can be used as is or after modifications for various applications such as antimicrobial treatments. Recently, the iron chelating activity of Otrf has been exposed to upsurge the stimulus by an inhibitor of AMPc ß-lactamase of some antibiotics which are efficient against most ß-lactamase-producing bacteria. Cefiderocol is a siderophore cephalosporin antibiotic that has recently been developed to combat a variety of bacterial pathogens, including ?-lactam and carbapenem-resistant organisms. OtrF is currently recommended for use in conjunction with antibiotics unless further studies increase the effectiveness of OtrF. These studies highlight the separation, purification of potential diversity in antimicrobial efficacy of Otrf.

Concentration of egg white antimicrobial and immunomodulatory proteins is related to eggshell pigmentation across traditional chicken breeds

Eggshell colour, quality, and biosafety of table eggs are of significant commercial interest. To date, there have been few studies investigating the relationship between eggshell pigmentation and internal egg quality in commercially bred birds. Moreover, the genetic basis and mechanisms behind the effects of extrinsic factors on deposition of antimicrobial compounds in egg white and eggshell pigments are not fully understood. In the present study, we evaluate the effect of chicken breed identity, eggshell pigmentation and the role of extrinsic factors (year and breeder identity) on variability in the concentrations of 2 major egg white antimicrobial proteins (AMPs), lysozyme (LSM), and ovotransferrin (OVOTR), across 23 traditional chicken breeds. We found that chicken breed identity and eggshell pigmentation explained most variability in the concentration of egg white LSM and OVOTR. Year and breeder identity were also significant predictors of egg white LSM and OVOTR variability, and showed selective effects on the deposition of both AMPs in egg white. We also documented a positive correlation between concentration of egg white LSM and eggshell cuticle protoporphyrin in tinted and dark brown eggs, but not in brown, white, and blue eggs. We assume that a combination of both intrinsic genetic and hormonally regulated extrinsic factors is responsible for this relationship and for the variability in egg white AMPs. In this study, we demonstrate the existence of a relationship between eggshell pigmentation and egg white AMPs content in the eggs of traditional chicken breeds that may advertise the egg's antimicrobial potential and biosafety. These findings provide novel insights into the relationship between eggshell pigmentation and egg internal quality and may stimulate the recovery and exploitation of traditional chicken breeds for egg production, where the demands for egg quality and biosafety, in conjunction with animal welfare, are a priority.

Assessment of the Allergenic Potential of the Main Egg White Proteins in BALB/c Mice.

This work aimed to assess the contribution of the major egg white proteins, ovalbumin, ovomucoid, and lysozyme, to the induction and elicitation of allergenic responses. For this purpose, BALB/c mice were orally administered either the individual egg allergens or a mixture of the three proteins in the same proportion, to evaluate their relative allergenicity avoiding their different abundance in egg white. Cholera toxin was used as a T helper 2 (Th2)-polarizing adjuvant. Ovomucoid and lysozyme triggered the most severe anaphylaxis reactions upon oral challenge. In comparison to ovalbumin and ovomucoid, lysozyme was a more active promotor of early immunoglobulin E and immunoglobulin G1 production and stimulated stronger Th2-biased responses from both mesenteric lymph node and spleen cells. These results indicate that lysozyme is highly immunogenic and should be considered as a major allergen, whose clinical usefulness in the diagnosis, prognosis, and therapeutic approaches of egg allergy deserves further consideration.

Antioxidant and anticancer effects of functional peptides from ovotransferrin hydrolysates.

Egg white is a good source for making functional peptides that can be used in the food industry. Ovotransferrin (OTF) is one of the major egg white proteins, with many functional properties, including antioxidant, antimicrobial and antiviral activities. However, enzymatic hydrolysis of ovotransferrin is known to further improve the functional activities of OTF. The aim of this study was to investigate the antioxidant and anticancer activities of functional peptides produced by two-step enzyme hydrolysis of OTF. OTF hydrolysates were prepared using promod 278P, thermolysin and a combination of the two enzymes. OTF and OTF hydrolysates showed strong antioxidant activities when analyzed using the DPPH assay. However, only OTF hydrolysates showed a strong free radical scavenging activity when NO- or ABTS-scavenging activity was measured. OTF hydrolysates showed stronger cytotoxic activities than the natural OTF against human cancer cell lines. Furthermore, OTF hydrolysates prepared with promod 278P + thermolysin combination showed the strongest cytotoxic activity, and their IC50 values were 0.79, 0.78, 0.92 and 0.78 mg mL-1 against AGS, LoVo, HT-29 and HeLa, respectively. These results indicated that the two-step enzyme hydrolysates of OTF have great potential for use as a food ingredient with antioxidant and anticancer activities. © 2017 Society of Chemical Industry.

Impact of the substrate viscosity, potentially interfering proteins and further sample characteristics on the ion exchange efficiency of tangential flow membrane adsorbers

The use of tangential flow membrane adsorbers is a highly recommendable option for chromatographic separations of particle containing substrates or substrates with high viscosities, like egg white. Due to the direction of the convective flow in this stationary phase, the sample is not required to be filtered. Our earlier study investigated the flow conditions of tangential flow membrane adsorbers. There, it became clear that the flow is based on a mix of convective and diffusive mass transfer. The diffusive component of the transport of the target proteins into the membrane structure depends on the viscosity of the medium according to the Stokes–Einstein equation. Therefore, it was the aim of the present study to assess the impact of viscosity on the ion exchange efficiency. Further, the amount of the main target protein of egg white, the bio-functional protein lysozyme, is relatively low compared to the other major egg white proteins. As a high amount of non-target proteins in the substrate can also interfere with the diffusional mass transfer of the target protein, it was an additional aim to understand the impact of the substrate composition on the lysozyme adsorption.It was shown that the effective diffusion coefficients (Deff) as well as the maximal binding capacities (qmax) strongly depend on the egg white viscosity. Applying natural egg white with a mean viscosity of 70mPas, a qmax of 0.14mgcm−2 with a Deff of 1.5×10‐7cm2s−1 were achieved. If the viscosity of egg white was reduced by means of a mechanical pre-treatment, qmax was increased to 0.49mgcm−2 with a Deff of 9.2×10−7cm2s−1. Consequently, the adsorption efficiency strongly depends on the substrate viscosity. Additionally, the impact of high amounts of non-target proteins in egg white on the adsorption of the target protein lysozyme was analyzed. It was demonstrated that the other egg white proteins strongly interfere the lysozyme binding to the membrane (qmax of 0.49mgcm−2 compared to 0.65mgcm−2). This is probably due to steric effects and interactions between the proteins. Finally, it can be concluded that diffusion is an important transport mechanism of tangential flow membrane adsorbers, and if this is disturbed by inappropriate conditions, the process works less well.

PH and heat induced structural changes of chicken ovalbumin in relation with antigenic properties

Ovalbumin is the major egg white protein known to induce allergic reactions in humans. A comprehensive evaluation of the structural and antigenicity features of ovalbumin subjected to different pH and heat treatments was performed by combining fluorescence spectroscopic measurements, ELISA and in silico prediction. The intrinsic fluorescence spectra indicated modification of the ovalbumin tertiary structure depending on pH and applied temperature. The heat treatment caused the alteration of ovalbumin structure, which exhibited gradual hydrophobic exposure. The in depths check of ovalbumin molecular model, after performing molecular dynamics simulations, indicated the slight transition toward a typical β-strand dominant structure with the temperature increase. Moreover the immunoenzymatic test was employed to estimate the effect of the pH and thermal treatment on the stability of ovalbumin epitopes. Only a 5.5% reduction of the residual antigenicity was observed when heat treating the ovalbumin samples at pH 7.0, whereas a significant reduction (over 82%) of the antigenicity was obtained at pH 9.5 and temperatures over 80°C. Both pH and thermal treatment affected the conformation of ovalbumin. The reduced recognition of the modified native ovalbumin by specific antibodies at alkaline pH is most probably a consequence of significant changes in the local conformation of the epitopes.

Comparative proteomic analysis of egg white proteins during the rapid embryonic growth period by combinatorial peptide ligand libraries

Egg white proteins provide essential nutrients and antimicrobial protection during embryonic development. Although various biological functions of major egg white proteins have been investigated via embryogenesis, understanding of global changes in low-abundance proteins has been limited. In the current study, a proteomic analysis of low-abundance egg white proteins was conducted using combinatorial peptide ligand libraries (CPLL), two-dimensional gel electrophoresis (2-DE), and matrix-assisted laser desorption/ionization-time-of-flight with two mass analyzers for tandem mass spectrometry (MALDI-TOF MS/MS) during the rapid embryonic growth period. Significant increases in the relative abundance of 88 protein spots (P ≤ 0.05), of which 47 spots were found to correspond to 10 proteins from 8 protein families were identified over 16 d incubation. During this developmental process, the protein concentration increased and the amount of albumin solid material decreased in the residual egg white. Clusterin precursors were observed over a wide range of pH values and the tenp protein increased continuously during embryonic development. Low-abundance proteins were identified in a comparison of optimal incubation conditions to the altered conditions of 2 control groups to better understand the function of these proteins in egg whites. Collectively, these findings provide insight into the supportive role of the egg white during embryonic development, enabling a broader understanding of chick embryogenesis.

The Family Secrets of Avian Egg-Specific Ovalbumin and Its Related Proteins Y and X.

The ovalbumin gene family in Gallus gallus is composed of three homologous genes located within a 46 kb locus on chromosome 2: ovalbumin, ovalbumin-related protein Y (OVAY), and ovalbumin-related protein X (OVAX) genes. The expression of these genes in hen oviduct is under estrogen control, but their relative hormonal responsiveness and subsequent protein concentration in egg, is distinctive. Interestingly, all three proteins lack the classical signal peptide for secretion. Ovalbumin, OVAX, and OVAY belong to the serine protease inhibitor (serpin) family whose members share a common tertiary structure. Ovalbumin and OVAX are one of the few members of this family that do not express any protease inhibition activity whereas OVAY has been predicted to be inhibitory, by comparison with the consensus sequence for inhibitory serpins. In contrast to ovalbumin and OVAY, OVAX interacts with heparin, a negatively charged glycosaminoglycan, via a positively charged domain exposed at the surface of the molecule. Ovalbumin is the major egg white protein and might be a source of amino acids for the developing embryo. The physiological function of OVAY is not known, but recent data have revealed a possible role of this protein in early embryonic development. Considering the antibacterial activities of OVAX, this protein might play a role in egg defense. This review sheds light on the expression, biochemistry, and structural specificities of these three highly similar paralogs. It gives new clues in favor of diverging functions, which are likely to have arisen by duplication events from a common ancestral gene.

Oral administration of hen egg white ovotransferrin attenuates the development of colitis induced by dextran sodium sulfate in mice.

Ovotransferrin (OVT), one of the major hen egg white proteins, was shown to possess antimicrobial and antioxidant activities in vitro. However, there is no information regarding the in vivo preventative effect in chronic inflammatory diseases such as inflammatory bowel disease (IBD). The aim of the present study is to evaluate the anti-inflammatory effects of OVT in a mouse model of dextran sodium sulfate (DSS)-induced colitis. OVT (50 or 250 mg/kg BW) was given orally for 14 days to female BALB/c mice, and 5% DSS (MW 36-50 kDa) was used to induce acute colitis (days 7-14) via drinking water. The current in vivo study demonstrated that OVT significantly reduced clinical signs, weight loss, shortening of the colon, and inflammatory cytokine markers of disease. The histopathological analysis of the colon revealed that OVT reduced histological scores. These results indicate that the use of OVT may be a potential promising candidate for the prevention of IBD.

Ovalbumin-related Protein X Is a Heparin-binding Ov-Serpin Exhibiting Antimicrobial Activities

Ovalbumin family contains three proteins with high sequence similarity: ovalbumin, ovalbumin-related protein Y (OVAY), and ovalbumin-related protein X (OVAX). Ovalbumin is the major egg white protein with still undefined function, whereas the biological activity of OVAX and OVAY has not yet been explored. Similar to ovalbumin and OVAY, OVAX belongs to the ovalbumin serine protease inhibitor family (ov-serpin). We show that OVAX is specifically expressed by the magnum tissue, which is responsible for egg white formation. OVAX is also the main heparin-binding protein of egg white. This glycoprotein with a predicted reactive site at Lys(367)-His(368) is not able to inhibit trypsin, plasmin, or cathepsin G with or without heparin as a cofactor. Secondary structure of OVAX is similar to that of ovalbumin, but the three-dimensional model of OVAX reveals the presence of a cluster of exposed positive charges, which potentially explains the affinity of this ov-serpin for heparin, as opposed to ovalbumin. Interestingly, OVAX, unlike ovalbumin, displays antibacterial activities against both Listeria monocytogenes and Salmonella enterica sv. Enteritidis. These properties partly involve heparin-binding site(s) of the molecule as the presence of heparin reverses its anti-Salmonella but not its anti-Listeria potential. Altogether, these results suggest that OVAX and ovalbumin, although highly similar in sequence, have peculiar sequential and/or structural features that are likely to impact their respective biological functions.

Separation of ovotransferrin from chicken egg white without using organic solvents

Ovotransferrin is one of the major egg white proteins that have antimicrobial activity as well as iron binding capability. The objective of this study was to develop a simple and easy method to separate ovotransferrin without using organic solvents. Egg white was separated from yolk, added in a 1:1 ratio to distilled water (DW), and then homogenized. The ovomucin in the diluted egg white was removed by centrifugation, adjusting the pH to 4.5 to 5.0. The resulting supernatant was added to different ratios of ammonium sulfate and citric acid, and then centrifuged after holding overnight at 4°C. The precipitant, which contains ovotransferrin, was dissolved in DW, and ovotransferrin was precipitated using different ratios of ammonium sulfate and citric acid. The precipitant collected after centrifugation was dissolved with DW and subjected to ultrafiltration to remove salts and concentrate the solution. The purity of the ovotransferrin was determined using SDS-PAGE, the protein identified using Western blot, and the estimated yield calculated by weighing the ovotransferrin after freeze drying. Over 85% purity and over 83% yield were obtained from the combinations of 5.0% (wt/vol) ammonium sulfate and 2.5% (wt/vol) citric acid followed by 2.0% (wt/vol) ammonium sulfate and 1.5% (wt/vol) citric acid. Activity of the ovotransferrin showed similar activity with previously separated ovotransferrin. However, this method is simpler and more cost effective than the previous method. The isolated ovotransferrin can be used as is or after modifications for various applications such as antimicrobial treatments, anticancer treatments, and iron-supplementing agents for humans.

Crystal Growth of Calcite Mediated by Ovalbumin and Lysozyme: Atomic Force Microscopy Study

Ovalbumin and lysozyme are two major egg white proteins and putatively related to the formation of the mammillary layer of eggshells. In this work, we have investigated their influences on the morphology and growth kinetics of hillocks at the molecular scale using fluid-cell atomic force microscopy. Our studies identified two roles for ovalbumin, favoring the formation of amorphous calcium carbonate–protein clusters on terrace surface and accelerating the step growth kinetics via reduction of the energy barrier for ion attachment to crystal steps. The two effects are intimately linked to the inherent characteristics of ovalbumin, i.e., being acidic and amphiphilic. In contrast, lysozyme as a basic protein did not induce the formation of any moldable transient phases. Instead, it interacted with step edges and pinned them, leading to step bunching and even step advancement stop at higher concentrations. These roles and their associated interactions on the molecular scale are related to the macroscopic feat...

Co-purification of chicken egg white proteins using polyethylene glycol precipitation and anion-exchange chromatography

Obtaining several proteins from chicken egg whites from a single extraction in a continuous process is imperative because there are potential applications for egg white proteins in the pharmaceutical and food industries. In this study, chicken egg whites were divided into four components using polyethylene glycol precipitation. All precipitates, except for ovomucin (Precipitate A), were further purified through Q Sepharose Fast Flow anion-exchange chromatography. Lysozyme, ovotransferrin, ovalbumin and ovoflavoprotein were purified from Precipitate B, Precipitate C and Supernatant D, respectively, which were determined to be 91.84%, 94.55%, 96.45% and 88.16% pure, respectively, by HPLC. The results showed that this method is feasible for the co-purification of major egg white proteins with high purity. This method is scientifically practical and easy to use and thus will suitable for industrial-scale production.

Effect of Shell Eggs Storage on Ovomucin Extraction

Ovomucin, one of the major egg white proteins, has wide potential applications in food, functional food and nutraceutical industries due to its unique physicochemical properties and bioactivities. A new two-step method was recently developed to prepare high purity ovomucin; in this study, effect of shell egg storage on ovomucin extraction was investigated. The composition of ovomucin extracts from egg white was determined by gel filtration chromatography. Both storage temperature and time could significantly (p < 0.05) affect the purity and yield of the ovomucin. After 9 weeks of storage at 4 °C, the content of the ovomucin in the extract decreased from 92.5% to 82.4%, and the yield of ovomucin decreased from 214 to 120 mg/100 g of egg albumen (egg white). After 5 weeks of storage at 22 °C, the content of ovomucin extract decreased from 92.5% to 73.0%, and the yield of ovomucin decreased from 214 to 112 mg/100 g of egg albumen. Increase in egg white pH during prolonged storage leading to degradation of ovomucin is very likely responsible for the decreased extractability of ovomucin.