• Title/Summary/Keyword: Gel Permeation Chromatography

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Characterization of 65 kD Protein in Latex Excreted from Euphorbia lathyris (Euphorbia lathyris에서 분비되는 Latex 65kD 단백질의 특성규명)

  • Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.319-323
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    • 2004
  • Soluble latex protein fraction excreted from Euporbia lathyris laticifer was resolved by 10% SDS-polyacrylamide gel electrophoresis to identify distinctively displayed latex major protein bands including ELp65, ELp55, ELp43, ELp32 and ELp23. Among them, ELp65 was purified by ammonium sulfate precipitation, gel permeation chromatography and ion exchange chromatography. Its N-terminal amino acid sequencing revealed its homology to the leading region of mature peptide of tomato p69a subtilisin-like protease, suggesting a certain role involved in plant defense system. In the analysis of Southern blot hybridization using PCR-amplified tomato p69a probe DNA, E. lathyris genome was suggested to have a gene family consisting of 3-5 gene members putatively encoding subtilisin-like proteases.

Separation and Purification of Angiotensin-I Converting Enzyme Inhibitory Peptides from Layer Hydrolysate (김 가수분해물로부터 Angiotensin-I Converting Enzyme저해 Peptide의 분리$\cdot$정제)

  • LEE Heon-Ok;KIM Dong-Soo;DO Jeong-Ryong;KWAN Dae-Young
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.2
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    • pp.164-172
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    • 2001
  • The angiotensin-I converting enzyme (ACE) inhibitors from laver hydrolysate was isolated. Among the 13 kinds of proteases, Maxazyme NNP was most effective for preparing the high ACE inhibitory compound. In extraction conditions of ACE inhibitory peptide from laver hydrolysate, ACE inhibitory activity of hydrolysate treated with diethylether for decolorization and that of $70\%$ ethanol soluble fraction among the different ethanol concentrations were higher than other preparations. Low molecular fraction less than 3,000 dalton of layer hydrolysate separated by ultrafiltration had the highest ACE inhibitory activity, for further separation of ACE inhibitory peptide from laver hydrolysate, gel filtration chromatography (Sephadex G-25), reverse-phase HPLC (ODS & Vydac C-18) and gel permeation chromatography (Superdex Peptide HR) were performed. The molecular mass of the ACE inhibitory peptide fractions of gel permeation chromatography determined by electrospray-mass spectrometer were 413.48 (S1O2V2V1P),346.86 (S1O2V2V2P) and 320.32 (S2O6V3V1P) dalton and their amino acid sequence were Val-Gln-Gly-Asn, Thr-Glu-Thr and Phe-Arg, respectively.

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Purification of carbosymethyl cellulase from hybrid between aspergillus niger and penicillium verruculosum

  • Yang, Young-Ki;Lee, Jung-Sup;Park, Hyung-Nam;Moon, Myung-Nim;Kim, Hong-Sub;Kim, Jong-Se;Lim, Chae-Young;Rhee, Young-Ha
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.90-94
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    • 1996
  • The carboxymethyl cellulase (CMCase) was purified from the induced culture filtrate of hybrid TAPW15703 between Aspergillus niger and penicillium verruculosum made by nuclear transfer. The enzyme was purified 80 fold with an overall yield 17% from the culture medium by ammonium sulfate fractionation, Sephadex G-75 gel permeation chromatography, and DEAE-ion exchange column chromatography. The molecular weight of the CMCase has estimated to be 32,000 daltons on SDS-polyacrylamide gel electrophoresis and Sephadex G-150 gel permeation chromatography. The purified enzyme functions optimally at pH 4.0 and 4$0^{\circ}C$ The Km value for carbosymethyl cellulose was 68 mM. The enzyme activity was increased by the presence of $Mg^{2+}$and Mn$^{2+}$.

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Silk Fibroin Microsphere and Its Characterization

  • Yeo Ju-Hong;Lee Gwang-Gil;Lee Yong-U;Gwon Hae-Yong;U Sun-Ok
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.56-56
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    • 2003
  • Using gel filtration chromatography (GFC), pure separation of high molecular silk fibroin was obtained and silk fibroin microsphere particles (SFMP) could be simply made by spray dryer method. Also, some of its physicochemical properties and morphology were investigated. The average molecular weight (Mw) of pure silk fibroin protein dissolved in calcium chloride is about 61, 500 g/mol as measured by gel permeation chromatography(GPC). (omitted)

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Chromatographic Techniques for the Isolation and Purification of Metalloporphyrins from Crude Asphalts (크로마토그래피를 이용한 아스팔트로 부터 금속 포르피린의 분리및 정제)

  • Woo Ki Chae
    • Journal of the Korean Chemical Society
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    • v.28 no.6
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    • pp.393-398
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    • 1984
  • Porphyrin-rich materials were obtained from some crude asphalts by gel permeation chromatography and silica gel chromatography. After extraction of each chromatographic fractions through alumina with pyridine, more concentrated metalloporphyrins were obtained. Demetallation of metalloporphyrins was possible without destroying porphyrin ring to provide different type of metal free porphyrins.

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Molecular Weight Distribution Characterization of Organics for the Dongbok Lake Water by Coagulation and Adsorption of Activated Carbon (동복 호소수의 응집침전 및 활성탄 흡착에 의한 용존유기물 분자량 분포 특성)

  • 정경훈;최형일
    • Journal of Environmental Science International
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    • v.7 no.1
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    • pp.104-111
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    • 1998
  • The Dongbok lake water before and after alum coagulation and activated carbon adsorption were analyzed in terms of organic contents, molecular weight distributuin (MWD), and UV-absorbance. Dissolved organic compounds in the Dongbok lake were fractionated into three molecular size classes by gel permeation chromatography. The fractionation was reasonably successful in isolating compounds with The bulk of the dissolved carbon was present in compounds of molecular weight in the range of 3,000~10,000. Alum coagulation preferentially treated molecules of high molecular weight, which has molecules larger than 10,000. The dissolved organic carbon (DOC) removal after activated carbon adsorption was high in the Fraction B , IR . The $A_{260}$/DOC ratio after alum and activated carbon treatment the Fraction II, III. This results suggest that the organics remaining after each treatment has a trihalomethane formation potential

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Analysis of Low Molecular Weight Collagen by Gel Permeation Chromatography

  • Yoo, Hee-Jin;Kim, Duck-Hyun;Park, Su-Jin;Cho, Kun
    • Mass Spectrometry Letters
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    • v.12 no.3
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    • pp.81-84
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    • 2021
  • Collagen, which accounts for one-third of human protein, is reduced due to human aging, and much attention is focused on making collagen into food to prevent such aging. Gel permeation chromatography with Reflective Index (RI) detection (GPC/RI) was chosen as the most suitable instrument to confirm molecular weight distribution, and we explored the use of this technique for analysis of collagen peptide molecular sizes and distributions. Data reliability was verified by matrix-assisted laser desorption/ionization coupled to time-of-flight (MALDI-TOF) mass spectrometric analysis. The data were considered meaningful for comparative analysis of molecular weight distribution patterns.

Purification of a major protein with physiological activities from Panax ginseng C. A. Meyer (고려인삼(Panax ginseng C. A. Meyer)에서 생리활성을 보이는 25 kDa 주요단백질 (GMP)의 분리정제)

  • Kwon, Taek-H.;Oh, Sei-R.;Park, H.;Kim, Kyung-H.
    • Applied Biological Chemistry
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    • v.41 no.6
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    • pp.410-413
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    • 1998
  • The major protein (GMP) from the roots of Panax ginseng C. A. Meyer was purified, using gel filtration and ion exchange chromatography followed by reversed-phase and ion exchange FPLC. Staining analysis indicated that the protein has a carbohydrate moiety, which was also shown by band shift experiments using various glycosidases. Electrophoretic and gel permeation studies showed that GMP has an apparent molecular weight of 63 kDa composed of possibly two subunits of 25 kDa containing carbohydrate moiety. GMP showed an anticomplementary activity on the hemolysis of red blood cells, which is a screening tool for inflammation mediator search.

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