• Title/Summary/Keyword: wheat gliadin

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Development and Characterization of Anti-gliadin Polyclonal Antibody in Wheat

  • Chang, Suk Joo;Hong, Byung Hee;Seo, Yang Weon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.4
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    • pp.339-344
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    • 1999
  • Immunological method has been applied in biochemical genetic analysis of seed storage proteins. We developed and characterized anti-gliadin polyclonal antibody (AGPab) specific to gliadin fractions whose quality and quantity were known to be associated with wheat end-use quality. Reactions of anti-gliadin polyclonal antibody (AGPab) to gliadin were linearly decreased as AGPab and antigen were diluted. Dot-blot and immunoblot assay showed that produced AGPab specifically reacted to gliadin and mainly $\alpha$-, $\beta$-, and ${\gamma}$-gliadin subunits. Enzyme-linked immuno- sorbent assay (ELISA) was applied for quantifi-cation of gliadins in Korean wheat cultivars and breeding lines by using AGPab. High reactions between AGPab and gliadins were found in wheat cultivars Olmil and Olgeurumil. Significant difference of optical densities for alcohol soluble proteins among crop species was found, as wheat showed the highest value (0.697) followed by rye (0.295), and barley (0.066).

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Genomics Approach to Identify the Cause of the Missing Omega-5 Gliadin Protein in O-Free Wheat

  • Lee, Yun Gyeong;Choi, Sang Chul;Kang, Yuna;Kang, Chon-Sik;Kim, Changsoo
    • Plant Breeding and Biotechnology
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    • v.6 no.4
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    • pp.413-425
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    • 2018
  • A previous work developed and identified a new omega-5 gliadin deficient wheat line named O-free by crossing Keumkang and Olgeuru, which is nutritionally quite meaningful in that omega-5 gliadin is one of the known wheat allergens. To verify the characteristics of the O-free, we performed RNA sequencing (RNAseq) analysis of the O-free and the two parent lines (Keumkang and Olgeuru). The results of the similarity analysis with the ESTs for gliadins and glutenins showed that the O-free ESTs had no similarity with the omega-5 gliadin sequences but had similarity to other gliadins and glutenins. Furthermore, mapping results between the raw RNAseq data from the O-free and the omega-5 gliadin sequence showed a clear deletion of the N-terminal sequences which are an important signature of omega-5 gliadin. We also designed specific PCR primers that could identify omega-5 gliadin in the genomic DNA. The results showed that no omega-5 gliadin fragments were detected in the O-free. According to these results, we confirmed that the deficiency of omega-5 gliadin in the O-free is not caused by post-transcriptional or post-translational regulations such as epigenetic phenomena but by a simple deletion in the chromosome. Furthermore, we showed that the low-molecular weight glutenin subunit (LMW-GS) gene in the O-free had a single nucleotide polymorphism (SNP) causing a premature stop codon, resulting in a truncated polypeptide. We expect that the O-free line may serve as an excellent source of wheat that could prevail in the hypo-allergen wheat market, which has recently gained interest world-wide.

Influence of Methylcellulose on Properties of Wheat Gliadin Film Cast from Aqueous Ethanol

  • Song, Yihu;Li, Lingfang;Zheng, Qiang
    • Food Science and Biotechnology
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    • v.18 no.4
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    • pp.910-916
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    • 2009
  • Present work was focused on the influence of methylcellulose (MC) on steady rheology of wheat gliadin solution and the properties of glycerol plasticized gliadin films. The presence of MC below 0.99 wt% improved viscosity and flow activation energy of the 10 wt% gliadin solution significantly. In the casting films containing 0.2 g glycerol/g dry protein, the MC component aggregated in the gliadin matrix. The blend films containing less than 7.7 wt% MC exhibited higher Young's modulus (E) and tensile strength (${\sigma}_b$) and lower elongation at break (${\epsilon}_b$) in comparison with the pure gliadin film, which was related to the intermolecular interaction between MC and gliadins, the brittle fracture of the aggregated MC component, and the increase in glass transition temperature ($T_g$) of the gliadin phase. Increasing MC content led to a slight increase in water vapor permeability (WVP) without significant influence on the moisture absorption (MA).

Studies on the Chemical Composition of Korean Wheat Flour Proteins (I). Purification of Gliadin Proteins and Amino Acid Composition (한국산 밀가루 단백질의 화학적 조성에 관한 연구 (제1보). Gliadin 단백질의 아미노산 조성 및 그의 정제)

  • Sung Hye Cho;Kim Jun Pyong
    • Journal of the Korean Chemical Society
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    • v.21 no.3
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    • pp.210-214
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    • 1977
  • We have investigated the amino acid composition of gliadin and glutenin from wheat flour Jang Kwang variety and have purified the main gliadin protein by Sephadex column. The results were obtained as follows: 1. Five bands of component for both gliadin and glutenin were found in disc electrophoretic gel column 2. The highest content of amino acid in gliadin and glutenin were glutamic acid and alanine was present in comparatively large amounts in glutenin, whereas proline was rich in gliadin 3. Main gliadin protein was purified by Sephadex G-150 and A-25 column chromatography and identified its purity by disc electrophoresis.

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Effects of Autoclave and Microwave Treatments on the Antigenicity of Gliadin in Medium Wheat Dough (가압가열 및 microwave에 의한 중력분 반죽 gliadin의 항원성 변화)

  • Kwak, Ji-Hee;Kim, Koth-Bong-Woo-Ri;Lee, Chung-Jo;Kim, Min-Ji;Kim, Dong-Hyun;SunWoo, Chan;Jung, Seul-A;Kim, Hyun-Jee;Choi, Jung-Su;Kim, Seong-Won;Ahn, Dong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.44 no.1
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    • pp.55-60
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    • 2012
  • The aim of this study was to determine the optimal physical treatment to reduce the antigenicity of gliadin in wheat dough. Medium wheat dough was treated with an autoclave (5, 10, 30, and 50 min at $121^{\circ}C$, 1 atm), a microwave (1, 5, and 10 min) or both (10, 30, and 50 min/5, 10 min). The proteins in the dough extracts were analyzed by SDSPAGE and the binding ability of anti-gliadin IgG to gliadin was examined by ci-ELISA and immunoblotting. Results showed that the ability of anti-gliadin IgG to bind to gliadin in wheat dough treated with an autoclave alone or in combination with a microwave was decreased. Especially, it declined to ~77% after autoclaving for 30 min and 35% after both autoclaving for 50 min and microwaving for 5 min. In addition, the intensity of gliadin bands in SDS-PAGE were weakened and anti-gliadin IgG did not recognize gliadin in immunoblotting. However, microwaving alone did not affect the antigenicity of gliadin in wheat dough. These results indicate that autoclaving may affect the reduction of the antigenicity of gliadin in medium wheat dough. Moreover, autoclaving in combination with microwaving is more effective for reducing the antigenicity of wheat dough.

Changes in Allergenicity of Gliadin in Strong Wheat Flour by Physical Treatments (물리적 처리에 의한 강력분 밀가루 Gliadin의 항원성 변화)

  • Kang, Bo-Kyeong;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Ahn, Na-Kyung;Choi, Yeon-Uk;Choi, Jung-Su;Choi, Ho-Duk;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.4
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    • pp.557-563
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    • 2014
  • This study was conducted to evaluate the effects of physical treatments on the antigenicity of gliadin in strong wheat flour. Strong wheat flour was treated with an autoclave (5, 10, 30, 50 min), a microwave (1, 5, 10 min), or both (10, 30, 50 min/ 5, 10 min), followed by SDS-PAGE, immunoblotting, and Ci-ELISA using anti-gliadin IgG. The results indicated that the binding ability of IgG to gliadin in strong wheat flour slightly decreased after autoclaving or autoclaving/microwaving. In particular, the binding ability was reduced to about 87% after autoclaving for 50 min and to 89% after autoclaving/microwaving (50/5 min). In addition, gliadin bands in the 50 min autoclaved group disappeared in both SDS-PAGE and immunoblotting. On the other hand, the antigenicity of gliadin was unaffected by microwaving alone. In conclusion, the results of this study suggest that autoclaving may reduce the antigenicity of gliadin in strong wheat flour.

Studies on the Chemical Composition of Korean Wheat Flour Proteins (II). Molecular Weight of the Main Gliadin Protein and Its Terminal Amino Acid Determination (한국산 밀가루 단백질의 화학적 조성에 관한 연구 (제2보) 주 Gliadin 단백질의 분자량 측정 및 말단아미노산의 검출)

  • Sung Hye Cho;Jun Pyong Kim
    • Journal of the Korean Chemical Society
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    • v.21 no.3
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    • pp.215-218
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    • 1977
  • We have investigated for amino acid composition, molecular weight and terminal amino acid of the main gliadin protein which was purified by Sephadex column. The results were obtained as follows: 1. The highest content of amino acid was glutamic acid for the main gliadin protein. 2. The molecular weight was estimated 60,200 ${\pm}$200 for the main gliadin protein. 3. Identified N-terminal and C-terminal amino acid of the main gliadin protein were phenylalanine and methionine, respectively.

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Genotype and Environment Effects on Gliadin Content and Polyphenol Oxidase Activity in Wheat

  • Seo, Yong-Weon;Park, Yong-Hack;Hong, Byung-Hee;Park, Moon-Woong;Nam, Jung-Hyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.1
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    • pp.38-43
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    • 2000
  • The environment in which a given genotype is grown may influence its grain quality characteristics. When varieties are $\times$ evaluated over numerous environments, a variety environment interaction usually is observed, but the relative magnitude of environmental(E), genetic(G), and G $\times$ E effects on quality is unclear. In order to determine relative contribution of genotype, environment, and G $\times$ E interaction to the variations observed in grain quality characteristics, 18 Korean wheat cultivars and experimental lines were evaluated in two environments in 1998 and 1999. Correlation coefficients between grain quality and agronomic characteristics were also estimated. The analysis of variance for the optical density obtained by reaction bet- ween gliadin and anti-gliadin polyclonal antibody (AGPab) indicated that gliadin content measured by Enzyme-Linked Immunosorbent Assay(ELISA) was significantly in- fluenced by environment and cultivar differences. The significant differences of year and year $\times$ location were also found. The ratio of the variances associated with environmental effects to the variances associated with genetic effect gave relatively greater influence of environmental factor on gliadin content. The different protein content from same genotype grown in different environment might be associated with degree of storage protein accumulations. Significant relationships between ELISA and protein content, yield, ten spike weight, and ten spike number were detected. Polyphenol oxidase (PPO) activity was significantly influenced by year, location, cultivar and year $\times$ location. The variance in grain PPO activities among growing years appeared larger than the variation produced by the cultivar examined. This suggested that the growing environment contributed more to variability in grain PPO concentration.

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Current status of wheat allergy research and prospect

  • Son, Jae-Han;Yoon, Young-mi;Cheong, Young-Keun;Park, Jong-Chul;Kim, Kyong-Ho;Kim, Bo-Kyeong;Kang, Chon-Sik
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.259-259
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    • 2017
  • Wheat is one of important food crop in the world. However, wheat has some negative things that are allergen. Wheat includes four different allergen kwon as alpha-, beta- and gamma gliadin and amylase inhibitor. Recently, people are interesting to gluten free food or low gliadin food. To study about wheat allergies and research trend is important for future research like as wheat breeding and allergy reduction technology. In this study, we analyzed that the reported on the research field of wheat allergy have been reported until now, and was to provide direction for future research. The analysis of 235 episodes of major papers published from 2007 to 2016. The last 5 years from 2012, wheat allergy appears increasing number of related papers. The ratio of national papers 13% and 60% were published by the United States and other European countries, respectively. The fields of wheat allergen-related preclinical technology and wheat allergen related to genome research for discovery technology were represented high rate by 58% and 26%, respectively. In the case of Korea, significantly account genetic and breeding areas. Recently, however, the research of glutenin protein which is closely linked to wheat allergy is in progress. So, we expected to increase wheat allergy of the research is in the future in Korea.

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Changes in Antigenicity of Gliadin from Medium Flour by Autoclave and Microwave Treatments (가압가열 및 Microwave 처리에 의한 중력분 Gliadin의 항원성 변화)

  • Kwak, Ji-Hee;Kim, Koth-Bong-Woo-Ri;Lee, Chung-Jo;Kim, Min-Ji;Kim, Dong-Hyun;SunWoo, Chan;Jung, Seul-A;Kang, Ju-Youn;Kim, Hyun-Jee;Choi, Jung-Su;Kim, Seong-Won;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1423-1429
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    • 2011
  • This study was conducted to evaluate the effect of physical treatments on the antigenicity of gliadin in medium wheat flour. The wheat flour was treated with an autoclave (5, 10, 30, 50 min), a microwave (1, 5, 10 min), and both (10, 30, 50 min/ 5, 10 min), and investigated by SDS-PAGE, immunoblotting and Ci-ELISA using anti-gliadin IgG. The results showed that the binding ability of anti-gliadin IgG to gliadin in wheat flour was slightly decreased when autoclaved or when autoclaved and microwaved. Especially, it was reduced to about 69% after autoclaving for 50 min and 73% after autoclaving for 50 min and microwaving for 5 min. In addition, gliadin bands in the 50 min autoclaved group disappeared in both SDS-PAGE and immunoblotting. On the other hand, the antigenicity of gliadin was unaffected by microwaving alone. Consequently, there were no considerable changes in using an autoclave alone or in combination with a microwave. These results suggest that autoclaving may affect the reduction of the antigenicity of gliadin in medium wheat flour.