• Title/Summary/Keyword: Allergenic proteins

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Expression and purification of Soybean $\beta$-Conglycinin from ($\beta$-Conglycinin의 대장균 발현과 정제)

  • 노영희
    • The Korean Journal of Food And Nutrition
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    • v.12 no.2
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    • pp.184-190
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    • 1999
  • Soybean protein consists of two major components $\beta$-conglycinin and glycinin which together consti-tute 70% of the total seed storage protein at maturity. $\beta$-Conglycinin is trimeric glycoprotein and for-med by the assembly of various combinations of three subunits $\alpha$,$\alpha$' and $\beta$ which have molecular weig-hts of 69,000, 72,000 and 42,000, respectively. Recently $\beta$-conglycinin was identified as powerful LDL lip-oprotein receptor activation hypercholesterolemia and major allergenic proteins. To investigate these reasons we constructed an expression system of cDNA encoding $\alpha$-subunit of $\beta$-conglycinin in Escherichia coli and purified the expressed protein. The pro-$\beta$-conglycinin synthesized in Escherichia coli BL 21 (DE3)comprised approximately 15% of the total bacterial proteins and the expressed protein are formed sol-uble and trimer such as native protein in Escherichia coli cells. The highly expressed protein was purified to homogeneity by salt precipitation with 20~40 % ammonium sulfate ion-exchange chromatography with Q-sepharose and hydrophobic column chromatography with Butyltoyopearl.

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Proteomic Approach of the Protein Profiles during Seed Maturation in Common Buckwheat (Fagopyrum esculentum Moench.)

  • Park, Min-Hwa;Shin, Dong-Hoon;Han, Myoung-Hae;Yun, Young-Ho;Bae, Jeong-Sook;Lee, Yun-Sang;Chung, Keun-Yook;Lee, Moon-Soon;Woo, Sun-Hee
    • Korean Journal of Plant Resources
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    • v.22 no.3
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    • pp.227-235
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    • 2009
  • Single seeds of common buckwheat cultivar Suwon No. 1 when subjected to SDS-PAGE revealed very high polymorphism. High variation existed for protein or protein subunits with molecular weight 54-47kDa, 45-25kDa and 16-11kDa. The electrophoregram showed variation for globulin as well as other protein fractions. About 300 proteins were separated by two-dimensional electrophoresis in common buckwheat (Fagopyrum esculentum Moench.) seed. Seed maturation is a dynamic and temporally regulated phase of seed development that determines the composition of storage proteins reserves in mature seeds. Buckwheat seeds from 5, 10, 15, 20, and 25 days after pollination and matured stage were used for the analysis. This led to the establishment of high-resolution proteome reference maps, expression profiles of 48 spots. It was identified 48 proteins from MALDI-TOF/MS analysis of wild buckwheat seed storage proteins. The 48 proteins were found identical or similar to those of proteins reported in buckwheat and other plants; it is belonging to 9 major functional categories including seed storage proteins, stress/defense response, protein synthesis, photosynthesis, allergy proteins, amino acid, enzyme, metabolism, and miscellaneous. It appears that the major allergenic storage protein separated played the important role in buckwheat breeding and biochemical characterization.

Identification and characterization of allergens of Chironomus fkavuoynys adults (Chironomidae, Diptera) in mice (노랑털깔따구(Chironomus flaviplumus) 성충의 알레르기 항원단백 분석)

  • 이한일;이상화
    • Parasites, Hosts and Diseases
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    • v.34 no.1
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    • pp.35-48
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    • 1996
  • Non-biting midges fchironomidae, Dipteral are one of the largest insect families, which are distributed worldwidely and are found in nearly all types of inland waters. They are known to be aggressive inhalant allergens which cause allergenic diseases. In this study, the crude antigens of Chironomus SavipLumn adults which are most widely distributed in Korea were extracted. and their allergens were analysed with the sera from experimentally sensitized mice. The mice were immunized with $1{\;}\mu\textrm{g}{\;}or{\;}10{\;}\mu\textrm{g}$ of the crude antigens, respectively, and the specific serum IgE levels were measured by both ELISA and passive cutaneous anaphylaxis (PCA) techniques. The highest levels of both total IgE and chironomid-specific IgE were found in the mouse sera obtained after 9 weeks of the first infection with $1{\;}\mu\textrm{g}$ crude antigen. The crude antigen was separated into 16-18 protein bands on gel by SDS-PAGE. The crude extract was assessed by SDS-PAGE/immunoblot analysis. One IgE-binding band (65 kDa) was detected by developing with colorimetric substrate, and 4 IgE-binding bands (65, 52, 35 and 25 kDa) by developing with CSPD chemiluminescent substrate. The SDS-PAGE gel of the crude extract of chironomid adults was equally cut into 30 pieces and each of them was eluted to isolate proteins by molecular weight, and the allergenicity of each eluate was assessed by applying P-K test on rats. Proteins of 65, 35 and 15 KDa showed the highest P-K titer (${\times}512$) which was 16 times higher than that of the crude extract (${\times32}$). The P-K titer of 52 kDa protein was also 4 times higher ($128{\times}$) than that of the crude extract, whereas the 25 kDa protein poorly responded, which seemed not antigenic. In conclusion, the present result in mice demonstrated that adults of Chironomus fcuiplumus, a predominent species in Korea, cause allergenic diseases and the main allergens are 65, 52, 35 and 15 kDa proteins, of which 65 kDa protein seems to be a main allergen.

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Overview of Milk Allergens and Allergic Reaction Reduction Methods (우유 알레르기의 특성 및 저감화 방법에 대한 고찰)

  • Kim, Ki-Hwan;Seol, Kuk-Hwan;Oh, Mi-Hwa;Park, Beam Young;Kim, Hyoun Wook
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.1
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    • pp.67-73
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    • 2013
  • Food allergy is defined as adverse reactions toward food mediated by aberrant immune mechanisms. Cow's milk allergy is one of the most common food allergies in childhood. This allergy is normally outgrown in the first year of life, however 15% of allergic children remain allergic. Cow's milk allergy seem to be associated with casein (${\alpha}_{s1}$-CN), ${\beta}$-lactoglobulin and whey protein. In addition to this, many other milk proteins are antigenic and capable of inducing immune responses. Various food processing affects the stability, structure and intermolecular interactions of cow milk proteins, as a result reduction the allergenic capacity. Heating, hydrolysis, chemical, proteolytic and other processes such as gamma-ray irradiation, high pressure, using probiotics treatments of milk to obtain hypoallergenic milk have been developed to reduce allergic reactions.

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Allergenicity assessment of novel proteins expressed in genetically modified organisms (유전자변형생물체의 알레르기성 평가와 이해)

  • Lee, Sang-Gu;Oh, Seon-Woo;Park, Soo-Yun;Park, Hyoun-Min;Kim, Eun-Ha;Jin, So-Ra;Ryu, Tae-Hun
    • Journal of Plant Biotechnology
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    • v.48 no.4
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    • pp.201-206
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    • 2021
  • To ensure the safety of developing or importing genetically modified organisms (GMOs), Korea has enacted the "LMO Act." Accordingly, the safety of using GMOs as food or feed is evaluated in accordance with the concept of "substantial equivalence" proposed by OECD. The allergenicity of GMOs is assessed as a part of their safety evaluation. The methods of allergenicity assessment have been discussed by various international organizations, such as the OECD, FAO, and WHO. The main methods used for the allergenicity assessment of proteins newly expressed in GMOs include assessment of the physicochemical stability of these proteins, evaluation of their amino acid homology with existing allergenic proteins, and serum screening. In this study, we describe guidelines and related studies for the allergenicity assessment of GM crops.

Allergenicity and toxicity evaluation of the PAT protein expressed in herbicide-tolerant genetically modified Zoysia japonica (제초제저항성 GM 잔디에서 발현된 PAT 단백질의 알레르겐 유발 가능성 및 독성 평가)

  • Jeong, Hye-Rin;Sun, Hyeon-Jin;Kang, Ji-Nam;Kang, Hong-Gyu;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.47 no.4
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    • pp.316-323
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    • 2020
  • This study aimed to evaluate the potential allergenicity and oral toxicity of the phosphinothricin acetyltransferase (PAT) protein expressed in Zoysia japonica, a herbicide-tolerant genetically modified (GM) zoysiagrass. In silico analysis of PAT showed no similarities with any known allergenic or toxic proteins, with <35% amino acid sequence homology with known allergens across a length of 80 amino acids and no continuous eight amino acid identity with known allergens. The PAT protein expressed in Z. japonica degraded very rapidly in the simulated gastric fluid in the presence of pepsin, and, no glycosylation of PAT was observed. The oral toxicity test revealed no mortality or toxic effect in mice following PAT administration at 4,000 mg/kg body weight. Our findings indicate that the PAT protein expressed in Zoysia japonica does not exhibit allergenic or toxic properties.

Food allergy

  • Han, Young-Shin;Kim, Ji-Hyun;Ahn, Kang-Mo
    • Clinical and Experimental Pediatrics
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    • v.55 no.5
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    • pp.153-158
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    • 2012
  • Food allergy is an important public health problem affecting 5% of infants and children in Korea. Food allergy is defined as an immune response triggered by food proteins. Food allergy is highly associated with atopic dermatitis and is one of the most common triggers of potentially fatal anaphylaxis in the community. Sensitization to food allergens can occur in the gastrointestinal tract (class 1 food allergy) or as a consequence of cross reactivity to structurally homologous inhalant allergens (class 2 food allergy). Allergenicity of food is largely determined by structural aspects, including cross-reactivity and reduced or enhanced allergenicity with cooking that convey allergenic characteristics to food. Management of food allergy currently focuses on dietary avoidance of the offending foods, prompt recognition and treatment of allergic reactions, and nutritional support. This review includes definitions and examines the prevalence and management of food allergies and the characteristics of food allergens.

ASSESSMENT OF STABILITY AND ALLERGENICITY OF FOOD ALLERGENIC PROTEINS

  • Lee, J-H;Yoon, W-K;Han, S-B;Yun, S-O;Park, S-H;Lee, H-J;Yoon, P-S;Moon, J-S;Kim, H-C
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.180-180
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    • 2002
  • The potential allergenicity of the transgene products in genetically modified organisms (GMOs), has been an important issue. As a part of the risk assessment of GMOs, we investigated the physicochemical stability and the immunogenicity of food allergens to determine their allergenicity.(omitted)

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A PCR Method for Rapid Detection of Buckwheat Ingredients in Food (식품에서 메밀 성분의 검출을 위한 PCR 방법)

  • Jeon, Young-Jun;Kang, Eun-Sil;Hong, Kwang-Won
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.276-280
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    • 2007
  • Buckwheat often causes severe allergic reactions in sensitive people. One of the major allergenic proteins in common buckwheat (Fagopyrum esculentum) has been found to be a BW10KD protein. In this study, we developed a PCR method to detect buckwheat ingredients in food using primers corresponding to the allergenic BW10KD gene. Five pairs of oligonucleotide primers successfully enabled PCR amplification of the specific regions of the genomic BW10KD DNA from buckwheat, but no amplification from seven other cereals and beans (barley, wheat, German millet, African millet, soybean, red bean, and black bean). The proposed PCR method was applied to analyze 12 processed foods (buckwheat flour, buckwheat noodle, buckwheat jelly, wheat noodle, instant noodle, black sesame gruels, sunsik, cookie, misutkaru, and three kinds of cereal); among them, only three samples including buckwheat flour, buckwheat noodle and buckwheat jelly showed a positive reaction to the detection. This PCR method was able to detect as little as 1 ng of common buckwheat DNA. This rapid and specific PCR method would be applicable to detect allergenic buckwheat ingredients in food.