• Title, Summary, Keyword: Ovalbumin

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Effect of Ultraviolet Irradiation on Molecular Properties of Ovalbumin (자외선 조사가 Ovalbumin의 분자적 성질에 미치는 영향)

  • Cho, Yong-Sik;Song, Kyung-Bin;Yamada, Koji;Han, Gui-Jung
    • Applied Biological Chemistry
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    • v.51 no.4
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    • pp.276-280
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    • 2008
  • To elucidate the effects of ultraviolet (UV) irradiation on molecular properties of ovalbumin, the molecular weight profile, secondary structure and tertiary structure of proteins were examined after irradiation by UV with 254 nm wavelength for 4, 8, 16 and 32 hrs, respectively. UV irradiation of protein solution caused the disruption on the native state of protein molecules. SDS-PAGE and gel permeation chromatography indicated that radiation caused initial fragmentation of polypeptide chains and as a result subsequent aggregation due to cross-linking of protein molecules. Circular dichroism (CD) study showed that UV irradiation caused the change on the secondary structure resulting in decrease of helical structure or compact denature on structure of protein depending on irradiation period. Fluorescence spectroscopy indicated that irradiation quenched the emission intensity excited at 280 nm. These results suggest that UV irradiation affect the molecular properties of ovalbumin and may have potential as a means to change the antigenicity of protein allergen.

Conformational Properties of Disulfide-Free Recombinant Chicken Ovalbumin

  • Jeoung, Yeon-Hee;Yu, Myeong-Hee
    • BMB Reports
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    • v.32 no.3
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    • pp.247-253
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    • 1999
  • Chicken egg ovalbumin is a non-inhibitory member of the serpin (serine protease inhibitors) family whose members share a common tertiary fold. In the present study, we succeeded in high-level production of a disulfide-free form of refolded recombinant ovalbumin. Conformational characterization of the recombinant ovalbumim revealed that it is well-folded, following two-state unfolding transition with the midpoint of transition at 4.7 M at $25^{\circ}C$. This value is very close to that of the reduced form of authentic ovalbumin. The recombinant ovalbumin can serve as a model molecule of non-inhibitory serpins in comparative studies with inhibitory members of the serpin family.

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Enzyme-Linked Immunosorbent Assay for Identification of Irradiated Eggs (효소면역 측정법에 의한 방사선 조사 계란의 검출)

  • 이경애;최윤정;양재승
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.6
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    • pp.1030-1034
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    • 2000
  • The ovalbumin, a most sensitive egg white protein to irradiation was purified from irradiated hen's eggs. Eggs were irradiated in their shells to 0~7 kGy. To investigate for a practical use in identifying of irradiated eggs, competitive ELISA using ovalbumin was peformed. The binding activity of ovalbumin to anti-ovalbumin IgG was reduced in a dose-dependent manner by irradiating up to 7 kGy, and consider-ably lowered after irradiating at 7 kGy. The concentration of 50% inhibition of ovalbumin to IgG was increased to 1.5~3.7 times in an irradiation dose-dependent relationship. SDS-PAGE of ovalbumin showed that the partial breakdown of ovalbumin was induced by irradiation. The lowering of binding activity was probably due to the partial breakdown of ovalbumin by irradiation. These results demonstrated that the ELISA should be quite useful and effective methods for the identification of irradiated eggs.

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Sequential Separation of Lysozyme and Ovalbumin from Chicken Egg White

  • Abeyrathne, Nalaka Sandun;Lee, Hyun Yong;Ahn, Dong Uk
    • Food Science of Animal Resources
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    • v.33 no.4
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    • pp.501-507
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    • 2013
  • Lysozyme was trapped from $2{\times}$ diluted egg white using Amberlite FPC 3500 ion exchange resin (1 g/10mL of egg white). The lysozyme bound to the resin was recovered using 0.1 N glycine-NaOH buffers, pH 9.0, containing 0.5 M NaCl. After separating lysozyme, the pH of the egg white solution was adjusted to 4.75 and centrifuged to remove interfering proteins. The supernatant was collected, added with 2.5% citric acid and 5.0% ammonium sulfate combination to precipitate egg white proteins, except for ovalbumin. After centrifugation, both supernatant (S1) and precipitant were collected. The precipitant was dissolved with 4 volumes of distilled water, and then 2.0% ammonium sulfate and 1.5% citric acid combinations added, stirred overnight in a cold room, and centrifuged. The resulting supernatant (S2) was pooled with the first supernatant (S1), desalted using an ultrafiltration unit, heat-treated at $70^{\circ}C$ for 15 min, and then centrifuged. The supernatant was collected as an ovalbumin fraction and lyophilized. The separated proteins were confirmed using Western blotting. The yield of lysozyme and ovalbumin was > 88.9% and > 97.7%, respectively, and the purity of lysozyme and ovalbumin was > 97% and 87%, respectively. The results indicated that the protocol was simple, and separated lysozyme and ovalbumin effectively.

Defining B Cell Epitopes of Ovalbumin for the C57BL/6 Mice Immunized with Recombinant Mycobacterium smegmatis

  • Kim, Hyo-Joon;Lee, Yang-Min;Hwang, Joon-Sung;Won, Ho-Shik;Kim, Bok-Hwan
    • BMB Reports
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    • v.32 no.5
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    • pp.461-467
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    • 1999
  • Recombinant Mycobacterium smegmatis expressing ovalbumin was used to immunize C57BL/6(H-$2^b$) mice, and the humoral immunity against recombinant ovalbumin was analyzed. Antibodies were purified by denatured ovalbumin-conjugated affinity chromatography. The epitopes of the antibodies were screened with a random peptide library displayed on the tip of fUSE5 filamentous phage pIII minor coat proteins. Two peptides, IRLADR and SPGAEV, were selected predominantly by the recognition of purified antibodies using biopanning methods. The composition of the peptide sequence with the primary structure of OVA revealed that the peptide sequence analogizes to INEAGR, part of the $^{323}ISQAVHAAHAEINEAGR^{339}$ sequence previously reported as the antigenic determinant for murine Band also Th cell epitopes (I-$A^d$ binding). Also, the structures of these mimotopes obtained from restrained molecular dynamic computations resulted in the formation of a $\beta$-turn proven to be a secondary structure of the parent peptide within the ovalbumin molecule, enabling us to confirm the structural similarity. This study demonstrates that immunization with recombinant M. smegmatis can generate neutralizing antibodies identical with those induced by the administration of natural antigenic proteins and supports the potential use of mycobacteria as vaccine delivery vehicles.

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Amelioration of Asthmatic-Related Symptoms by an Aqueous Extract of Angelica archangelica L. (신선초의 물 추출물에 의한 천식 증상의 감소)

  • Heo, Jin-Chul;Lee, Sang-Han
    • Journal of Life Science
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    • v.18 no.10
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    • pp.1336-1341
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    • 2008
  • Inflammation through the respiratory tract is a crucial event in immune disorders, including asthma, and atopic rhinitis. To investigate whether an aqueous extract of Angelica archangelica L. (AaL) has a beneficial influence in terms of anti-asthmatic activity, its effects on an ovalbumin-induced asthmatic model were examined. Mice sensitized to ovalbumin were orally administered the AaL extract, and their lungs examined by Haematoxylin-Eosin staining to determine IL-4/13 cytokine expression. The AaL extract exerted strong anti-asthmatic effects by regulating each level in the $CD4^+$ cell number, IL-4/13, and other target markers in the lungs. Together, these results collectively indicate that the aqueous AaL extract ameliorates asthmatic symptoms effectively in a mouse ovalbumin-challenge model.

Development of the Purification Method of Ovotransferrin in Egg White (난백 내 Ovotransferrin의 분리방법에 관한 연구)

  • Jang, A.;Jo, Y.J.;Lee, M.;Kim, J.C.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.1025-1032
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    • 2005
  • This study was carried out to separate ovotransferrin in chicken egg white by gel chromatography and heparin affinity chromatography. In gel filtration which was performed with 50mM Phosphate buffer (pH 7.2, 0.15M salt) at a flow rate of 2.0 ml/min, ovotransferrin and ovalbumin were eluted together in fraction number 11-16. In order to separate pure ovotransferrin, fraction No. 12-14 of them which have high concentration of ovotransferrin were concentrated and rechromatographed. However, the ovotransferrin did not separated clearly. In heparin affinity chromatography, the separation was performed with 50mM ethylaminetetraacetic acid (EDTA, pH7.2) and 50mM Phosphate buffer (pH 7.2, 0.15M salt contained) on ferrous and ferric ion saturated column at as same flow rate as gel filtration system's. Ovotransferrin and albumin were eluted together at 10-15min (fraction No.3) and 15-20min (fraction No.4), respectively. However, purified ovotransferrin was eluted at 156-165min and 165-175min (tube No.32-33) with 50 mM phosphate buffer (pH 7.2, 0.15M salt free), respectively. Heparin affinity chromatography with ferric ion saturated column was resulted in the best separation of ovotransferrin rather than separation by gel chromatography and ferrous ion saturated heparin affinity chromatography.

Oral Administration Effects of Herbal Extracts on Atopic Dermatitis in Balb/c Mice Sensitized by Ovalbumin (Ovalbumin으로 유발된 아토피피부염 모델 마우스에 대한 복합한약추출물 경구투여의 효과)

  • Kim, Kyung-Jin;Kim, Gyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.27 no.3
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    • pp.72-83
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    • 2014
  • Objective: This study was to evaluate whether herbal mixture (HM-A : Houttuynia cordata Thunberg, Rubus coreanus, Rehmannia glutinosa, Prunus yedoensis, HM-B : Houttuynia cordata Thunberg, Rubus coreanus, Rehmannia glutinosa, Angelica gigas nakai) supresses the development of atopic dermatitis in Balb/c mice sensitized by ovalbumin. Methods: Mice were sensitized by intraperitoneal injection of ovalbumin plus aluminum hydroxide hydrate, followed by epicutaneous sensitization for 6 weeks. After induced atopic dermatitis, HM-A and HM-B were orally administrated for two weeks(once a two days) as a 50 mg/kg concentration. After all mice were sacrificed at the end of the experiment, skin and blood were harvested. Results: Oral administration group was reduced the infiltration of eosinophils, mast cells and total T cells on the skin areas as well as blood analysis. Also, cutaneous expression of IL-4,13,17 decreased. Blood IgE level was decreased. Conclusion: These drugs could be potential candidates for the atopic dermatitis.

Ultrasonic Velocity and Absorption Mesurements in Gel of Proteins (초음파에 의한 단백질 gel화의 연구)

  • 김정구
    • Proceedings of the Acoustical Society of Korea Conference
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    • pp.29-34
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    • 1998
  • Egg white의 gel화에 따른 음속과 흡수의 변화가 60와 75$^{\circ}C$에서 크게 나타난 것이 egg white의 어느 단백질 성분에 의한 것인가를 조사하기 위해 egg white의 주요한 단백질 성분인 obalbumin, conalbumin, ovomucoid protein에 대해 gel화에 따른 음속 및 흡수의 변화를 온도 10-95$^{\circ}C$의 범위에서 초음파pulse법을 사용하여 측정하였다. Ovalbumin는 7$0^{\circ}C$, conalbumin는 5$0^{\circ}C$에서 gel화가 시작되었고 ovomucoid는 측정온도범위내에서는 gel화가 진행되지 않았다. Gel화하는 이상의 온도에서 음속과 흡수에 대하여 aging측정을 행하여 gel화에 의한 dam속과 흡수의 변화를 관측하였다. 그 결과 conalbumin는 5$0^{\circ}C$, ovalbumin는 75$^{\circ}C$에서 음속과 흡수의변화가 많이 일어났다. Egg white의 60와 75$^{\circ}C$의 gel화에 의한 음속과 흡수의 큰 변화는 각각 conalbumin과 ovalbumin에 의한 것임을 알았고 Conalbumin과 ovalbumin는 aging 온도를 parameter로하여 이력현상이 관측되었다.

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Fermented red ginseng and ginsenoside Rd alleviate ovalbumin-induced allergic rhinitis in mice by suppressing IgE, interleukin-4, and interleukin-5 expression

  • Kim, Hye In;Kim, Jeon-Kyung;Kim, Jae-Young;Han, Myung Joo;Kim, Dong-Hyun
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.635-644
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    • 2019
  • Background: To increase the pharmacological effects of red ginseng (RG, the steamed root of Panax ginseng Meyer), RG products modified by heat process or fermentation have been developed. However, the antiallergic effects of RG and modified/fermented RG have not been simultaneously examined. Therefore, we examined the allergic rhinitis (AR)-inhibitory effects of water-extracted RG (wRG), 50% ethanol-extracted RG (eRG), and bifidobacteria-fermented eRG (fRG) in vivo. Methods: RBL-2H3 cells were stimulated with phorbol 12-myristate-13-acetate/A23187. Mice with AR were prepared by treatment with ovalbumin. Allergic markers IgE, tumor necrosis factor-${\alpha}$, interleukin (IL)-4, and IL-5 were assayed in the blood, bronchoalveolar lavage fluid, nasal mucosa, and colon using enzyme-linked immunosorbent assay. Mast cells, eosinophils, and Th2 cell populations were assayed using a flow cytometer. Results: RG products potently inhibited IL-4 expression in phorbol 12-myristate-13-acetate/A23187-stimulated RBL-2H3 cells. Of tested RG products, fRG most potently inhibited IL-4 expression. RG products also alleviated ovalbumin-induced AR in mice. Of these, fRG most potently reduced nasal allergy symptoms and blood IgE levels. fRG treatment also reduced IL-4 and IL-5 levels in bronchoalveolar lavage fluid, nasal mucosa, and reduced mast cells, eosinophils, and Th2 cell populations. Furthermore, treatment with fRG reduced IL-4, IL-5, and IL-13 levels in the colon and restored ovalbumin-suppressed Bacteroidetes and Actinobacteria populations and ovalbumin-induced Firmicutes population in gut microbiota. Treatment with ginsenoside Rd significantly alleviated ovalbumin-induced AR in mice. Conclusion: fRG and ginsenoside Rd may alleviate AR by suppressing IgE, IL-4, IL-5, and IL-13 expression and restoring the composition of gut microbiota.