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The Biochemical Journal Feb 19711. Purified ovomucin was isolated as an insoluble glycoprotein complex from thick egg white. 2. A homogeneous glycoprotein, designated alpha-ovomucin, of molecular...
1. Purified ovomucin was isolated as an insoluble glycoprotein complex from thick egg white. 2. A homogeneous glycoprotein, designated alpha-ovomucin, of molecular weight 210000 and containing N-acetylglucosamine (6.7%, w/w), N-acetylgalactosamine (0.6%, w/w), galactose (1.8%, w/w), mannose (4.6%, w/w), N-acetylneuraminic acid (1.0%, w/w) and sulphate (0.7%, w/w), was isolated from preparations of reduced ovomucin by sedimentation equilibrium in a density gradient of caesium chloride formed in the presence of 4m-guanidine hydrochloride. 3. A carbohydrate-rich fraction, designated beta-ovomucin (which is homogeneous by sedimentation-velocity analysis in 5m-guanidine hydrochloride but which is heterogeneous by analytical sedimentation equilibrium in a density gradient of caesium chloride in the presence of 4m-guanidine hydrochloride), containing N-acetylglucosamine (11.0%, w/w), N-acetylgalactosamine (8.7%, w/w), galactose (19.2%, w/w), mannose (4.1%, w/w), N-acetylneuraminic acid (13.8%, w/w) and sulphate (2.7%, w/w), was also obtained from preparations of reduced ovomucin by the density-gradient method. 4. Mild acid hydrolysis of the unfractionated ovomucin complex showed that N-acetylneuraminic acid occupied a terminal position of the oligosaccharide chains. 5. Alkaline beta-elimination reactions with the unfractionated ovomucin complex indicated that N-acetylgalactosamine was linked by alkali-labile bonds to hydroxy amino acids.
Topics: Amino Acids; Centrifugation, Density Gradient; Chromatography, Gel; Chromatography, Paper; Egg White; Electrophoresis; Galactosamine; Galactose; Glucosamine; Glycoproteins; Hexosamines; Hexoses; Hydrolysis; Mannose; Models, Structural; Molecular Weight; Neuraminic Acids; Oxidation-Reduction; Sulfates
PubMed: 5119791
DOI: 10.1042/bj1210537 -
Journal of Functional Biomaterials Apr 2024As an essential nutrient, lutein (LUT) has the ability to aid in the prevention of eye diseases, cardiovascular diseases, and cancer. However, the application of LUT is...
Natural Biomolecule Ovomucin-Chitosan Oligosaccharide Self-Assembly Nanogel for Lutein Application Enhancement: Characterization, Environmental Stability and Bioavailability.
As an essential nutrient, lutein (LUT) has the ability to aid in the prevention of eye diseases, cardiovascular diseases, and cancer. However, the application of LUT is largely restricted by its poor solubility and susceptibility to oxidative degradation. Thus, in this study, LUT-loaded nanogel (OVM-COS-LUT) was prepared by a self-assembly of ovomucin (OVM) and chitosan oligosaccharide (COS) to enhance the effective protection and bioavailability of LUT. The nanogel had excellent dispersion (PDI = 0.25) and an 89.96% LUT encapsulation rate. XRD crystal structure analysis confirmed that the encapsulated LUT maintained an amorphous morphology. In addition, the nanogel showed satisfactory stability with pH levels ranging from 2 to 9 and high ionic strengths (>100 mM). Even under long-term storage, the nanogel maintained an optimistic stabilization and protection capacity; its effective retention rates could reach 96.54%. In vitro, digestion simulation showed that the bioaccessibility and sustained release of OVM-COS-LUT nanogel was superior to that of free LUT. The nanogel provided significant antioxidant activity, and no significant harmful effects were detected in cytotoxicity analyses at higher concentrations. In summary, OVM-COS-LUT can be utilized as a potential safe oral and functional carrier for encapsulating LUT.
PubMed: 38667568
DOI: 10.3390/jfb15040111 -
Poultry Science Apr 2018Egg white contains many functionally important proteins: ovalbumin (54%), ovotransferrin (12%), ovomucoid (11%), ovoglobulin (G2 and G3, 8%), ovomucin (3.5%), and... (Review)
Review
Egg white contains many functionally important proteins: ovalbumin (54%), ovotransferrin (12%), ovomucoid (11%), ovoglobulin (G2 and G3, 8%), ovomucin (3.5%), and lysozyme (3.5%) are major proteins, while ovoinhibitors, ovomacroglobulin, ovoglycoprotein, ovoflavoprotein, thiamine-binding proteins, and avidin are minor proteins present in egg white. These proteins, as well as the peptides derived from the proteins, have been recognized for their functional importance as antioxidant, antimicrobial, metal-chelating, anti-viral, anti-tumour, and angiotensin-converting enzyme (ACE)-inhibitory activities. Among the functional properties of the peptides, antioxidant and antimicrobial activities are important characteristics for food processing while other properties such as ACE-inhibitory activity of the peptides can have important health-related functionalities. Bioactive peptides can be produced from egg white proteins by enzyme hydrolysis, chemical treatments, or thermal treatments at different pH conditions. The effective functional peptides produced from egg white proteins are usually smaller than 2 kDa in molecular size. However, these peptides are known for their beneficial activities in vitro only, and little work has been done to prove their beneficial effects in vivo. Therefore, further studies are needed to see if the bioactive peptides derived from egg white proteins are helpful for humans in the future.
Topics: Angiotensin-Converting Enzyme Inhibitors; Animals; Antioxidants; Chickens; Egg Proteins
PubMed: 29340654
DOI: 10.3382/ps/pex399 -
International Archives of Allergy and... 2022Molecular studies of hen's egg allergens help define allergic phenotypes, with IgE to sequential (linear) epitopes on the ovomucoid (OVM) protein associated with a...
INTRODUCTION
Molecular studies of hen's egg allergens help define allergic phenotypes, with IgE to sequential (linear) epitopes on the ovomucoid (OVM) protein associated with a persistent disease. Epitope profiles of other egg allergens are largely unknown. The objective of this study was to construct an epitope library spanning across 7 allergens and further evaluate sequential epitope-specific (ses-)IgE and ses-IgG4 among baked-egg reactive or tolerant children.
METHODS
A Bead-Based Epitope Assay was used to identify informative IgE epitopes from 15-mer overlapping peptides covering the entire OVM and ovalbumin (OVA) proteins in 38 egg allergic children. An amalgamation of 12 B-cell epitope prediction tools was developed using experimentally identified epitopes. This ensemble was used to predict epitopes from ovotransferrin, lysozyme, serum albumin, vitellogenin-II fragment, and vitellogenin-1 precursor. Ses-IgE and ses-IgG4 repertoires of 135 egg allergic children (82 reactive to baked-egg, the remaining 52 tolerant), 46 atopic controls, and 11 healthy subjects were compared.
RESULTS
183 peptides from OVM and OVA were screened and used to create an aggregate algorithm, improving predictions of 12 individual tools. A final library of 65 sequential epitopes from 7 proteins was constructed. Egg allergic children had higher ses-IgE and lower ses-IgG4 to predominantly OVM epitopes than both atopic and healthy controls. Baked-egg reactive children had similar ses-IgG4 but greater ses-IgE than tolerant group. A combination of OVA-sIgE with ses-IgEs to OVM-023 and OVA-028 was the best predictor of reactive phenotype.
CONCLUSION
We have created a comprehensive epitope library and showed that ses-IgE is a potential biomarker of baked-egg reactivity.
Topics: Allergens; Animals; Chickens; Egg Hypersensitivity; Epitopes; Female; Humans; Immunoglobulin E; Immunoglobulin G; Ovalbumin; Ovomucin; Peptides; Vitellogenins
PubMed: 34818647
DOI: 10.1159/000519618 -
Foods (Basel, Switzerland) Feb 2023In this study, differences in the protein content and functional and physicochemical properties of four varieties of egg white (EW) were studied by adding 4-10% sucrose...
In this study, differences in the protein content and functional and physicochemical properties of four varieties of egg white (EW) were studied by adding 4-10% sucrose or NaCl and then heating them at 70 °C for 3 min. According to a high-performance liquid chromatography (HPLC) analysis, the percentages of ovalbumin, lysozyme and ovotransferrin rose with an increase in the NaCl or sucrose concentration; however, the percentages of ovomucin and ovomucoid decreased. Furthermore, the foaming properties, gel properties, particle size, α-helixes, β-sheets, sulfhydryl groups and disulfide bond content also increased, whereas the content of β-turns and random coils decreased. In addition, the total soluble protein content and functional and physicochemical properties of black bone (BB) chicken and Gu-shi (GS) EWs were higher than those of Hy-Line brown (HY-LINE) and Harbin White (HW) Ews ( < 0.05). Subsequently, transmission electron microscopy (TEM) confirmed the changes in the EW protein structure in the four varieties of Ews. As the aggregations increased, the functional and physicochemical properties decreased. The protein content and functional and physicochemical properties of Ews after heating were correlated with the concentration of NaCl and sucrose and the EW varieties.
PubMed: 36832956
DOI: 10.3390/foods12040881 -
BMC Genomics Aug 2006Mucins are large glycoproteins that cover epithelial surfaces of the body. All mucins contain at least one PTS domain, a region rich in proline, threonine and serine.... (Comparative Study)
Comparative Study
An inventory of mucin genes in the chicken genome shows that the mucin domain of Muc13 is encoded by multiple exons and that ovomucin is part of a locus of related gel-forming mucins.
BACKGROUND
Mucins are large glycoproteins that cover epithelial surfaces of the body. All mucins contain at least one PTS domain, a region rich in proline, threonine and serine. Mucins are also characterized by von Willebrand D (VWD) domains or SEA domains. We have developed computational methods to identify mucin genes and proteins based on these properties of the proteins. Using such methods we are able to characterize different organisms where genome sequence is available with respect to their mucin repertoire.
RESULTS
We have here made a comprehensive analysis of potential mucins encoded by the chicken (Gallus gallus) genome. Three transmembrane mucins (Muc4, Muc13, and Muc16) and four gel-forming mucins (Muc6, Muc2, Muc5ac, and Muc5b) were identified. The gel-forming mucins are encoded within a locus similar to the corresponding human mucins. However, the chicken has an additional gene inserted between Muc2 and Muc5ac that encodes the the alpha-subunit of ovomucin, a protein similar to Muc2, but it is lacking a PTS domain. We also show that the beta-subunit of ovomucin is the orthologue of human MUC6. The transmembrane Muc13 gene is in chicken as well as in mammals adjacent to the HEG (heart of glass) gene. HEG has PTS, EGF and transmembrane domains like Muc13, suggesting that these two proteins are evolutionary related. Unlike previously known mucins, the PTS domain of Muc13 is encoded by multiple exons, where each exon encodes a repeat unit of the PTS domain.
CONCLUSION
We report new mucin homologues in chicken and this information will aid in understanding the evolution of mucins in vertebrates. The fact that ovomucin, a protein not found in mammals, was located in the same locus as other gel-forming mucins provides strong support that these proteins are evolutionary related. Furthermore, a relationship of HEG and the transmembrane Muc13 is suggested on the basis of their biochemical properties and their presence in the same locus. Finally, our finding that the chicken Muc13 is distributed between multiple exons raises the interesting possibility that the length of the PTS domain could be controlled by alternative splicing.
Topics: Amino Acid Sequence; Animals; Chickens; Computational Biology; Evolution, Molecular; Exons; Forecasting; Genome; Humans; Membrane Glycoproteins; Molecular Sequence Data; Mucin 5AC; Mucin-2; Mucin-5B; Mucin-6; Mucins; Multigene Family; Ovomucin; Phylogeny; Protein Structure, Tertiary; Sequence Homology, Amino Acid; Tandem Repeat Sequences; Vertebrates; Zebrafish; Zebrafish Proteins
PubMed: 16887038
DOI: 10.1186/1471-2164-7-197 -
Poultry Science Apr 2014Ovotransferrin and ovomucoid were separated using 2 methods after extracting the ovotransferrin- and ovomucoid-containing fraction from egg white. Diluted egg white...
Ovotransferrin and ovomucoid were separated using 2 methods after extracting the ovotransferrin- and ovomucoid-containing fraction from egg white. Diluted egg white (2×) was added to Fe(3+) and treated with 43% ethanol (final concentration). After centrifugation, the supernatant was collected and treated with either a high-level ethanol (61% final concentration) or an acidic salt combination (2.5% ammonium sulfate and 2.5% citric acid) to separate ovotransferrin and ovomucoid. For the high-level of ethanol method, ovotransferrin was precipitated using 61% ethanol. After centrifugation, the precipitant was dissolved in 9 vol. of distilled water and the residual ethanol in the solution was removed using ultrafiltration. The supernatant, mainly containing ovomucoid, was diluted with 4 vol. of water, had ethanol removed, and was then concentrated and used as the ovomucoid fraction. For the acidic salt precipitation method, the ethanol in the supernatant was removed first. The ethanol-free solution was then concentrated and treated with a 2.5% ammonium sulfate and 2.5% citric acid combination. After centrifugation, the precipitant was used as the ovotransferrin and the supernatant as the ovomucoid fraction. The ovomucoid fraction from both of the protocols was further purified by heating at 65°C for 20 min and the impurities were removed by centrifugation. The yields of ovomucoid and ovotransferrin were >96 and >92%, respectively. The purity of ovomucoid was >89% and that of the ovotransferrin was >88%. The ELISA results confirmed that the activity of the separated ovotransferrin was >95%. Both of the protocols separated ovotransferrin and ovomucoid effectively and the methods were simple, fast, and easy to scale up.
Topics: Animals; Blotting, Western; Chemical Precipitation; Chickens; Conalbumin; Egg White; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Ethanol; Food Handling; Ovomucin
PubMed: 24706979
DOI: 10.3382/ps.2013-03649 -
Organic & Biomolecular Chemistry Sep 2018Antimicrobial and anti-proliferative meleagrin and oxaline are roquefortine C-derived alkaloids produced by fungi of the genus Penicillium. Tandem O-methylations...
Antimicrobial and anti-proliferative meleagrin and oxaline are roquefortine C-derived alkaloids produced by fungi of the genus Penicillium. Tandem O-methylations complete the biosynthesis of oxaline from glandicoline B through meleagrin. Currently, little is known about the role of these methylation patterns in the bioactivity profile of meleagrin and oxaline. To establish the structural and mechanistic basis of methylation in these pathways, crystal structures were determined for two late-stage methyltransferases in the oxaline and meleagrin gene clusters from Penicillium oxalicum and Penicillium chrysogenum. The homologous enzymes OxaG and RoqN were shown to catalyze penultimate hydroxylamine O-methylation to generate meleagrin in vitro. Crystal structures of these enzymes in the presence of methyl donor S-adenosylmethionine revealed an open active site, which lacks an apparent base indicating that catalysis is driven by proximity effects. OxaC was shown to methylate meleagrin to form oxaline in vitro, the terminal pathway product. Crystal structures of OxaC in a pseudo-Michaelis complex containing sinefungin and meleagrin, and in a product complex containing S-adenosyl-homocysteine and oxaline, reveal key active site residues with His313 serving as a base that is activated by Glu369. These data provide structural insights into the enzymatic methylation of these alkaloids that include a rare hydroxylamine oxygen acceptor, and can be used to guide future efforts towards selective derivatization and structural diversification and establishing the role of methylation in bioactivity.
Topics: Imidazoles; Methyltransferases; Models, Molecular; Ovomucin; Penicillium; Protein Conformation
PubMed: 30141817
DOI: 10.1039/c8ob01565a -
Jornal de Pediatria 2020To assess the frequency of baked egg tolerance in IgE-mediated egg allergy patients through the oral food challenge and to assess the tolerance predictability of...
OBJECTIVE
To assess the frequency of baked egg tolerance in IgE-mediated egg allergy patients through the oral food challenge and to assess the tolerance predictability of different skin prick tests, as well as specific serum IgE measurement to egg proteins.
METHODS
In this cross-sectional study, 42 patients with a diagnosis of egg allergy were submitted to different skin prick tests with egg (in natura, boiled, muffin, ovalbumin, and ovomucoid), and specific IgE to egg white, ovalbumin, and ovomucoid; as well as to the oral food challenge with food containing egg, extensively baked in a wheat matrix.
RESULTS
Of the total, 66.6% of patients tolerated the ingestion of egg-containing foods in the oral food challenge. A comparative analysis with positive and negative oral food challenge found no significant differences regarding age, gender, other food allergies, or even specific skin prick tests and IgE values between the groups.
CONCLUSIONS
The study demonstrated an elevated frequency of baked egg food-tolerant individuals among egg allergy patients. None of the tested markers, skin prick tests, or specific IgE, were shown to be good predictors for identifying baked egg-tolerant patients. The oral food challenge with egg baked in a matrix is central to demonstrate tolerance and the early introduction of baked foods, improving patients' and families' quality of life and nutrient intake.
Topics: Allergens; Cooking; Cross-Sectional Studies; Egg Hypersensitivity; Eggs; Humans; Immune Tolerance; Immunoglobulin E; Ovomucin; Quality of Life; Skin Tests
PubMed: 31513760
DOI: 10.1016/j.jped.2019.08.002 -
Scientific Reports Apr 2016The CRISPR/Cas9 system is a simple and powerful tool for genome editing in various organisms including livestock animals. However, the system has not been applied to...
The CRISPR/Cas9 system is a simple and powerful tool for genome editing in various organisms including livestock animals. However, the system has not been applied to poultry because of the difficulty in accessing their zygotes. Here we report the implementation of CRISPR/Cas9-mediated gene targeting in chickens. Two egg white genes, ovalbumin and ovomucoid, were efficiently (>90%) mutagenized in cultured chicken primordial germ cells (PGCs) by transfection of circular plasmids encoding Cas9, a single guide RNA, and a gene encoding drug resistance, followed by transient antibiotic selection. We transplanted CRISPR-induced mutant-ovomucoid PGCs into recipient chicken embryos and established three germline chimeric roosters (G0). All of the roosters had donor-derived mutant-ovomucoid spermatozoa, and the two with a high transmission rate of donor-derived gametes produced heterozygous mutant ovomucoid chickens as about half of their donor-derived offspring in the next generation (G1). Furthermore, we generated ovomucoid homozygous mutant offspring (G2) by crossing the G1 mutant chickens. Taken together, these results demonstrate that the CRISPR/Cas9 system is a simple and effective gene-targeting method in chickens.
Topics: Animals; Avian Proteins; CRISPR-Cas Systems; Cells, Cultured; Chick Embryo; Chickens; Female; Gene Targeting; Germ Cells; Male; Mutagenesis; Mutation; Ovalbumin; Ovomucin; Reproducibility of Results
PubMed: 27050479
DOI: 10.1038/srep23980