• Title/Summary/Keyword: storage proteins

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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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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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Milk Protein-Stabilized Emulsion Delivery System and Its Application to Foods

  • Ha, Ho-Kyung;Lee, Won-Jae
    • Journal of Dairy Science and Biotechnology
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    • v.38 no.4
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    • pp.189-196
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    • 2020
  • Milk proteins, such as casein and whey protein, exhibit significant potential as natural emulsifiers for the preparation and stabilization of emulsion-based delivery systems. This can be attributed to their unique functional properties, such as the amphiphilic nature, GRAS (generally recognized as safe) status, high nutritional value, and viscoelastic film-forming ability around oil droplets. In addition, milk protein has been used as a coating material in emulsion-based delivery systems to protect bioactive compounds during food processing and storage owing to its unique functional properties. These properties include the ability to bind lipophilic bioactive compounds and antioxidant activity. In this review, we present the use of milk proteins as emulsifiers for the formation of emulsions and food applications of milk protein-stabilized emulsion delivery systems.

Analysis of Protein Function and Comparison on Expression of Protein in Taekwang During Maturation using Proteomic Techniques (Proteomics를 이용한 등숙기 차이에 따른 콩 종실 저장단백질 발현양상 비교 분석)

  • Cho, Seong-Woo;Kim, Tae-Sun;Kwon, Soo-Jeong;Roy, Swapan Kumar;Lee, Chul-Won;Kim, Hong-Sig;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.41-46
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    • 2015
  • In the present study, different expression of protein from Taekwang was revealed by 2-DE, and expressions of protein on each week after flowering was investigated. After analysis of expression of protein, MALDI-TOF was executed to identify expected protein function. Results revealed that there were three patterns of expression of protein during the maturing. The first pattern was that proteins were gradually expressed as up-regulation from 1 week to 6 week. The second pattern was that proteins were expressed gradually from 1 week to 5 week and then it started down-regulation in 6 week. The last pattern was that proteins were gradually as up-regulation from 1 week to 3 week and then down-regulation until 6 week. This phenomenon suggests that young stage has more protein related to correspondence mechanism against disease and growth and then maturing stage has more expression of protein related to storage protein. In MALDI-TOF analysis, p24 oleosin isoform A protein was identified that relates oleosin which is synthetic product in oil body. This protein spot increased gradually until 5 week and then decreased after 5 week. It explained that the protein is active until maturing stage to protect oil in seed and then its activity has gradually degraded. This result may be expected that a protein, related to growth of a seed has increased until maturing and then a seed fills up with a storage protein.

Column cleaning, regeneration and storage of silica-based columns (실리카 기반 컬럼의 세척, 재생 및 보관 가이드)

  • Matt James;Mark Fever
    • FOCUS: LIFE SCIENCE
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    • no.1
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    • pp.1.1-1.4
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    • 2024
  • This article provides comprehensive guidance on the maintenance, cleaning, regeneration, and storage of silica-based HPLC (High-Performance Liquid Chromatography) columns. The general considerations emphasize the importance of using in-line filters and guard cartridges to protect columns from blockage and irreversible sample adsorption. While these measures help, contamination by strongly adsorbed sample components can still occur over time, leading to an increase in back pressure, loss of efficiency, and other issues. To maximize column lifetime, especially with UHPLC (Ultra-High Performance Liquid Chromatography) columns, it is advisable to use ultra-pure solvents, freshly prepared aqueous mobile phases, and to filter all samples, standards, and mobile phases. Additionally, an in-line filter system and sample clean-up on dirty samples are recommended. However, in cases of irreversible compound adsorption or column voiding, regeneration may not be possible. The document also provides specific recommendations for column cleaning procedures, including the flushing procedures for various types of columns such as reversed phase, unbonded silica, bonded normal phase, anion exchange, cation exchange, and size exclusion columns for proteins. The flushing procedures involve using specific solvents in a series to clean and regenerate the columns. It is emphasized that the flow rate during flushing should not exceed the specified limit for the particular column, and the last solvent used should be compatible with the mobile phase. Furthermore, the article outlines the storage conditions for silica based HPLC columns, highlighting the impact of storage conditions on the column's lifetime. It is recommended to flush all buffers, salts, and ion-pairing reagents from the column before storage. The storage solvent should ideally match the one used in the initial column test chromatogram provided by the manufacturer, and column end plugs should be fitted to prevent solvent evaporation and drying out of the packing bed.

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Characterization of 27K Zein as a Transmembrane Protein

  • Lee, Dong-Hee
    • BMB Reports
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    • v.31 no.2
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    • pp.196-200
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    • 1998
  • Zeins, maize storage proteins, are retained in the endoplasmic reticulum (ER) during the subcellular targeting process without the ER retention signal. Circumstantial data indicate that the 27K zein is an ER transmembrane protein. The potential transmembrane domain may permit the 27K zein to remain in the ER. This study investigated the potential transmembrane feature by employing alkaline extraction, proteinase K digestion, and surface biotinylation on isolated intact protein bodies. These assays consistently support the possibility of the 27K zein as a transmembrane protein. The 27K zein polypeptide was shown to be associated with alkali-stripped membranes. The polypeptide was digested by proteinase K to a smaller fragment. According to surface biotinylation, the 27K zeins was labeled to the exclusion of other classes of zeins. This study, therefore, concludes that the 27K zein has an ER transmembrane domain, which may serve as an anchor for zeins' ER retention.

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Characterization of 7S and 11S Globulins in Soybean Varieties Differing in Seed Size and Their Effects on the Properties of Soybean Curd

  • Kim, Sun-Lim;Koo, Han-Mo;Chun, Se-Cheol;Kim, Jung-Tae;Kim, Min-Young;Chi, Hee-Youn;Kim, Eun-Hye;Kim, Hyun-Bok;Kim, Mi-Jung;Seo, Bo-Ram;Kang, Eun-Young;Seo, Su-Hyun;Chung, Ill-Min
    • Food Science and Biotechnology
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    • v.17 no.1
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    • pp.135-143
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    • 2008
  • The ratio between 11S Sand 7S globulins was greater in large seed size varieties (LSSVs) of soybean than in small seed size varieties (SSSVs) or medium seed size varieties (MSSVs) but did not differ between MSSVs and SSSVs. The cysteine and methionine contents of 11S globulins were greater than those of total seed proteins and 7S globulins. The acidic monoamino monocarboxylic amino acids were the most abundant class of amino acid in soybean seed (27.2%) and soybean curd protein (29.3%). Isolated 7S and 11S fractions were analyzed by HPLC. Of the 12 peaks detected, 4 constituted 64.1% of the proteins of the SSSVs, 65.6% of the proteins of the MSSVs, and 70.5% of the proteins of the LSSVs. The 11S/7S globulin ratio was related to the yield and hardness of soybean curd. The MSSVs had the greatest yield of soybean curd, but the soybean curd hardness of the MSSVs was greater than that of the SSSVs. These results show that the 11S/7S ratio and color of soybean seeds can be used to predict the yield, hardness, and color of soybean curd.