• Title/Summary/Keyword: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Separation and Purification of Endo-polygalacturonase from Korean Jujube (한국산 대추로부터 Endo-polygalacturonase 분리 및 정제)

  • Choi, Cheong;Chun, Sung-Sook;Cho, Young-Je;Woo, Heui-Seob;Kim, Tae-Wan;Heo, Young-Hoon
    • Applied Biological Chemistry
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    • v.37 no.4
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    • pp.243-247
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    • 1994
  • Endo-polygalacturonase was purified from Jujube. The purification procedures included DEAE-cellulose ion exchange chromatography and gel filtration on Sephdex G-100. Enzyme was purified as a single protein band and purification yield was about 6%. When the purified enzyme was applied to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight was estimated about 19,000. Purified enzyme formed hexagonal board type.

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Proteins and Amino Acid Composition of Korea Ginseng Classified by Years (한국인삼의 연근별 단백질 및 아미노산 조성)

  • Choi, Cheong;Yoon, Sang-Hong;Bae, Man-Jong;An, Bong-Jean
    • Korean Journal of Food Science and Technology
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    • v.17 no.1
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    • pp.1-4
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    • 1985
  • For the systematic investigation of biochemical characteristics of Korean ginseng protein by years, protein fractions were analyzed by the techniques of polyacrylamide gel electrophoresis, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration, while the amino acid composition was studied by amino acid autoanalyzer. Results of polyacrylamide gel electrophoresis and SDS-PAGE showed a few difference in pattern and number of bands depending on the age of the root. However, the number of bands obtained from polyacrylamide gel electrophoresis and SDS-PAGE was 8 and 7 to 11, respectively. When water extracted proteins were fractionated by Sephadex G-200, the main peak among 2 peaks was collected and lyophillized. Its mol. wt. was extimated to be 43,000 by the SDS-PAGE method. In amino acid composition of water extracted protein and main fraction of gel filtration, arginine content was the highest, 47.17% in water extracted protein and 57.36% in main fraction followed by glutamic acid and asparatic acid. On the contrary, cystine and methionine contents were very low in both cases.

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The enhancement of protein separation by duplex SDS-PAGE (Duplex SDS-PAGE를 이용한 단백질 분리향상)

  • Pyo, Jae Sung;Roh, Si Hun;Song, Jin-Su;Lee, Kyung Hyeon;Kim, Hie-Joon;Park, Jeong Hill;Kwon, Sung Won
    • Analytical Science and Technology
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    • v.19 no.6
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    • pp.529-534
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    • 2006
  • The protein separation with molecular weight using SDS-PAGE(sodium dodecyl sulfate-polyacrylamide gel electrophoresis) is the one of the most conventional and simple techniques. In, this study, two dimensional SDS-PAGE using same separation principle consecutively was investigated and compared with one dimensional SDS-PAGE. The enhanced separation from duplex SDS-PAGE was observed and separated proteins in the gel were identified by MALDI TOF MS. Identified proteins from different gel spots were found to have different gi numbers. Therefore, duplex SDS-PAGE separation method will be used for economic separation method in the future because only tiny amount of inexpensive reagents are used to perform duplex SDS-PAGE.

Solubility and Electrophoretic pattern of Korea Ginseng Protein (한국산 인삼 단백질의 용해성 및 전기영동 패턴)

  • Choi, Cheong;Yoon, Sang-Hong;Bae, Man-Jong;An, Bong-Jeon
    • Applied Biological Chemistry
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    • v.28 no.2
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    • pp.88-91
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    • 1985
  • For the systematic investigation of biochemical characteristics of Korean ginseng protein, protein fractions were analyzed by the techniques of sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. The effect of pH and various salts on extractibility of ginseng protein were determined while the amino acid composition was studied by amino acid autoanalyzer. The protein was consisted of 66.08% of albumin and 20.51% of glutelin. Extractability of ginseng protein was the lowest in pH 3.0 and the highest in $pH\;6.0{\sim}8.0$. Among the neutral salts solution, $0.4M\;Na_2CO_3$ showed maximum extractability while $1.0M\;MgSO_4$ solution showed the least extractability. Resonable precipitation was obtained by 40% of acetone and ammonium sulfate. It has been shown by SDS polyacrylamide gel electrophoresis that the soluble protein had 11 bands. The molecular weight for the main protein of the soluble protein wasestimated to be 43,000. In amino acid composition of water extracted protein, arginine content was the highest 47.17% while on the contray, proline and cystine contents were very low.

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Production and Purification of Alkaline Protease from Streptomyces sp. (Streptomyces속 균주가 생성하는 Alkaline Protease의 생산 및 정제)

  • Choi, Cheong;Chung, Yung-Gun;Sung, Sam-Kyung;Choi, Kwang-Soo;Lee, Jae-Sung;Cho, Young-Je;Kwon, Oh-Jin
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.169-177
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    • 1992
  • An alkaline protease producing microorganism was isolated from soil and identified as Streptomyces griseus HC-1141. The optimum culture condition of Streptomyces griseus HC- 1141 for the production of alkaline protease was as follows; 0.5% casein, 0.05% ammonium chloride, 0.1% ferrous sulfate. 2.0% lactose, pH 8.0 and 84 hrs. The enzyme was purified about 53 folds by ammonium sulfate treatment, DEAE-cellulose ion exchange chromatography and gel filtratioo on Sephadex G-150. The homogeneity of the purified enzyme was verified by polyacrylamide gel electrophoresis. The molecular weight was estimated to be 31,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis. This enzyme consists of glycine and glutamic acid as major amino acids. The N-terminal and C-terminal residues of the alkaline protease were leucine and histidine respectively.

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Production and Purification of Alkaline Lipase from Serratia sp. AL-11 (Serratia sp. AL-11이 생산하는 Alkaline Lipase의 생산 및 정제)

  • Choi, Cheong;Kim, Tae-Wan;Cho, Young-Je
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.695-701
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    • 1995
  • An alkaline lipase producing bacteria was isolated from soil and identified as Serratia liquefaciens AL-11. from the results of analysis of its morphological, biochemical and physiological properties. This strain showed the highest productivity of alkaline lipase when grown at pH 9.0 and 30$\circ$C for 42 hours in the medium of 1% peptone, 0.5% tryptone, 0.9% yeast extract, 1% starch, 1% tween 80, 0.05% CaCl$_{2}$ and 0.05% NaCl. The enzyme was purified by ammonium sulfate treatment, Sephadex G-100 gel filtration and DEAE-Sephadex A-50 column chromatography. The specific activity of the purified enzyme was 27 unit/mg protein and the yield of enzyme activity was 61.3%. The homogeneity of the purified enzyme was verified by polyacrylamide gel disc electrophoresis. Molecular weight of the purified enzyme was estimated about 53,000 by sodium dodecyl sulfate- polyacrylamide gel electrophoresis. This enzyme is composed of 17 amino acids of which glycine, proline and glutamic acid were three miajor acids.

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$^{1}$H-NMR spectroscopic evidence on the glycosidic linkages of the transglycosylated products of low-molecular-weight $\beta$-D-glucosidase from trichoderma koningii (Trichoderma koningii에서 분비되는 .$\beta$-D-glucosidase의 반응산물에 대한 핵자기공명분석)

  • 이헌주;정춘수;강사욱;하영칠
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.35-42
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    • 1989
  • The mode of transglycosylation reaction observed during the action of low-molecular-weigh $\beta$-D-glucosidase ($\beta$-D-glucoside glucohydrolase, EC3.2.1.21) purified from Trichoderma koningii ATCC 26113 was investigated using $^{1}H$-NMR spectroscopy. The enzyme was purified by the series of procedures including ammonium sulfate precipitation, and fractionations by column chromatographies on Bio-Gel P-150, DEAE-Sephadex A-50, and SP-Sephadex C-50. The final purification was performed by the band eluation after preparative polyacrylamide gel electrophoresis. The enzyme showed its molecular size of 78,000 through the analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and its isoelectric point of 5.80 through the analysis of analytical isoelectric focusing. The H-1 proton resonances were analyzed. After the reaction of the enzyme with cellobiose, the reaction products were separated by high performance liquid chromatography using refractive index detector. H-1 resonances of the products were consisted with those of gentiobiose [$\beta$-D-glucopyranosyl--(1,6)-D-glucopyranose], and cellotriose [$\beta$-D glucopyranosyl-(1,4)-$\beta$-D-glucopyranosyl]-(1,4)-D-glucopyranose] with minor resonances of sophorose [$\beta$-D-glucopyranosyl-(1,2)-D-glucopyranose], respectively.

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Studies on the Composition of Protein and Lipid from Korean Walnut (Juglans regia L.) (한국산 호도의 단백질 및 지질의 조성에 관한 연구)

  • Choi, Cheong;Sung, Tae-Soo;Cha, Woen-Suep;Son, Cyu-Mok
    • Applied Biological Chemistry
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    • v.29 no.3
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    • pp.318-323
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    • 1986
  • We empolyed gel filtraction, polyacrylamide gel electrophoresis, amino acid autoanalyzer, thin layer chromatography for determining protein and lipid composition in walnut. The walnut contained 22.18% of crude protein and 64.23% of crude lipid. Glutamic acid (38.60%) was the major amino acid in soluble protein, followed by arginine and aspartic acid. The sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed 12 band in soluble protein of walnut, and collection rate of main protein fraction purified by Sephadex G-150 was 60.67%. The molecular weight for the main protein was estimated to be 43,000. The lipid fraction obtained by silicic acid column chromatography were mainly composed of about 93.05% neutral lipid, whereas compound lipid was only 7.0% level. Among the neutral lipid by thin layer chromatography, triglyceride was 82.05%, sterol ester and free fatty acid were 3.86% and 4.80%, repectively. The predominant fatty acids of total and neutral lipids were linoleic acid $(64.48{\sim}69.98%)$ and oleic acid $(13.89{\sim}15.36%)$. The major fatty acids of triglyceride separated from neutral lipid were linolenic acid (69.98%).

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Genetic Variation of High Molecular Weight Glutenin (HMW-Glu) Subunit in Korean Wheat

  • Hong, Byung-Hee;Park, Chul-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.4
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    • pp.259-263
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    • 1998
  • High molecular weight glutenin (HMW-Glu) subunit compositions of 73 Korean wheat cultivars and experimental lines were evaluated by using one dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This method is suitable for obtaining a good resolution of 1Dx2 and 1Ax2$^*$ without adverse effects on separation of other HMW-Glu subunits. Korean wheats examined in this study could be divided into 15 different groups on the basis of HMW-Glu subunit compositions. From the wheat lines tested, it was identified that there were three alleles at the Glu-Al, five at the Glu-Bl and three at the Glu-D1 loci. The null allele of the Glu-Al was occurred in high frequency (79.4%), while low frequencies for 1Ax1 (12.3%) and 1Ax2$^*$(8.2%) were found. High frequency (75.3%) of the subunit pairs of 1Bx7+1By8 at the Glu-Bl loci compared with other subunits was found. The frequencies of subunits 1Dx2. 2+1Dy12 and 1Dx2+1Dy12 from the Glu-D1 loci were 54. 8% and 37.0%, respectively. However, a few Korean wheat lines (8.2%) carried 1Dx5 + 1Dy10 subunit pair which are responsible for good breadmaking quality. The information of HMW-Glu subunit compositions provide a useful tool to characterize wheat lines, and can be directly used in selection of breeding lines of different end-use properties.

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Characterization of the Physical Form of Allergenic Cry j 1 in the Urban Atmosphere and Determination of Cry j 1 Denaturation by Air Pollutants

  • Wang, Qingyue;Morita, Jun;Gong, Xiumin;Nakamura, Shinichi;Suzuki, Miho;Lu, Senlin;Sekiguchi, Kazuhiko;Nakajima, Takuya;Nakajima, Daisuke;Miwa, Makoto
    • Asian Journal of Atmospheric Environment
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    • v.6 no.1
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    • pp.33-40
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    • 2012
  • In this study, we characterized the physical form of allergenic Cry j 1 in the urban atmosphere. Through an immunofluorescence antibody method, we showed that allergenic Cry j 1 exists as fine particles (${\leq}1.1{\mu}m$). To determine Cry j 1 concentrations and its particle size distribution, we used the ELISA method to confirm that most Cry j 1 exists as fine particles in the urban atmosphere and is found at high concentrations on fine day next to rainy day. Furthermore, we evaluated Cry j 1 denaturation by using the Biacore J system based on the surface plasmon resonence (SPR) principle and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). We showed that the dissociation constant ($K_D$) of Cry j 1 that has been exposed to urban polluted air is lower ($1.76{\times}10^{-14}$ M) than that of Cry j 1 ($1.32{\times}10^{-9}-3.37{\times}10^{-9}$ M) of original pollen grains that has not been exposed to air pollutants. Cry j 1 turns into low molecular weight proteins by reacting with various acidic solutions. In sum, we showed that allergenic Cry j 1 exists as fine particles that can deposit in the lower respiratory tract. This finding clarifies the relationship between Japanese cedar pollinosis and air pollutants.