• 제목/요약/키워드: Partial purification

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Partial Purification and Characterization of Fibrinolytic Substance from Wooltalikong (Phaseolus ssp.)

  • Oh Hae-Sook;Kim Jun-Ho
    • 대한의생명과학회지
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    • 제10권4호
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    • pp.415-420
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    • 2004
  • Fibrinolytic substance was purified from the Wooltalikong (Phaseolus ssp.), using DEAE-cellulose chromatography, Sephadex G-150 gel-filtration, and FPLC gel-filtration. The substance has a molecular weight of 5262.70 Da as measured by MALD-TOF mass spectrometry. It has a pH optimum at pH 6.0. The fibrinolytic activity of purified substance was inhibited by EDTA and 1,10-phenanthroline and slightly decreased by PMSF and pepstatin A. It shows the maximum fibrinolytic activity at 40℃ and the substance was stable up to 50℃. The activity of the substance was increased by Zn/sup 2+/ and was totally inhibited by Hg/sup 2+/. This study revealed that Wooltalikong could be a good source of fibrinolytic products due to its small molecular size and heat-resistant ability.

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한국 전통 발효식품 중의 Ethyl Carbamate 정량 (Fermentation Specific Carcinogen Ethyl Carbamate in Korean Traditional Foods)

  • 정현정;권훈정
    • 동아시아식생활학회지
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    • 제7권1호
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    • pp.41-46
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    • 1997
  • Diet is generally accepted as one of the important factors in human cancer development. Ethyl carbamate has been associated with cancer for several decades and mainly found in the fermented beverages and foodstuff. The relationship between ethyl carbamate and the human health cannot be ignored especially in the areas where fermented foods consists of regular food consumption. To investigate the ethyl carbamate exposure level in Korean population, commercial fermented food samples were collected form local markets I Seoul area and home-made varieties were collected throughout the country. Following partial purification the concentration of ethyl carbamete was determined by GC/ MS. The concentration of ethyl carbamate ranged to 70 ppb in soysauce, to 10 ppb in soybean paste, and to 5 ppb in vinegars. Korean traditional alcoholic beverages showed small amount of ethyl carbamate. The estimated daily exposure of Korean population was 0~1900ng/day. It would be prudent to put efforts to minimize the formation of ethyl carbamate since the risk from the higher range exposure cannot be ignored.

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인체에서 분리된 Lactobacillus acidophilus가 생산하는 박테리오신의 선별과 정제 (Screening and Partial Purification of Bacteriocins by Strains of Lactobacillus acidophilus Isolated from Human Origin)

  • 김세헌;김영교
    • Journal of Dairy Science and Biotechnology
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    • 제15권1호
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    • pp.21-26
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    • 1997
  • Lactobacillus acidophilus 223, 606, and NCFM-F among 21 isolated from fecal contents of humans demonstrated inhibitory activity attributed to bacteriocin(s). The bacteriocin(s) were heat stable and nondialyzable proteinous compounds and exhibited narrow inhibitory spectra of activity. Neither hydrogen peroxide nor pH were responsible for inhibitory action. All of the producer strains were resistant to their own bacteriocin(s). The bacteriocin(s) were purified by ammonium sulfate precipitation, gel chromatography and ion exchange chromatography for further characterization. The bacteriocin(s) of human origin exhibited similar characteristics.

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Partial Purification and Characterization of Thermostable Esterase from the Hyperthermophilic Archaeon Sulfolobus solfataricus

  • Chung Young Mi;Park Chan B.;Lee Sun Bok
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제5권1호
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    • pp.53-56
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    • 2000
  • A thermostable esterase from the hyper thermophilic archaeon Sulfolobus solfataricus was partially purified 590-fold with $16.2\%$ recovery. The partially purified esterase had a specific activity of $29.5\;{\mu}mol\;min^{-1}mg^{-1}$ when the enzyme activity was determined using p-nitrophenyl butyrate as a substrate. The apparent molecular weight was about 100 kDa, while the optimum temperature and pH for esterase were $75^{\circ}C$ and 8.0, respectively. The enzyme showed high thermal stability and solvent tolerance in comparison to its mesophilic counterpart. The enzyme also showed chiral resolution activity for (S)-ibuprofen, indicating that S. solfataricus esterase can be used for the production of commercially important chiral drugs.

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Saccharomyces cerevisiae에서 PNP의 부분 정제와 특성 (Partial Purification and Characterization of Purine Nucleoside Phosphorylase in Saccharomyces cerevisiae)

  • 최혜선
    • 미생물학회지
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    • 제29권3호
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    • pp.172-178
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    • 1991
  • Intracellular purine nucleoside phosphorylase (PNP) from Saccharomyces cerevisiae was partially purified using ammonium sulfate fractionation, heat treatment, a DEAE-Sephadex A-50 anion exchange chromatography and a Sephadex G-100 gel filtration chromatography. The enzyme was purified 20 fold with 3% recovery. The stability of enzyme was kept by addition of inosine and dithiothreitol. The pH optimum was found to be from 6.3 to 7.3 PNP was sensitive to 10mM of $Hg^{2+}$ , $Cu^{2+}$ , and was inactivated completely by 2 mM of p-chloromercuribenzoate and 5,5'-dithiobis (2-nitrobenzoate). The enzyme was capable of catalyzing the phosphorolysis of inosine, deoxyinosine, guanosine, deoxyguanosine and adenosine.

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고려홍삼 성분이 암독소 호르몬-L의 체지방 분해작용에 미치는 영향 (Effect of Korean Red Ginseng Component on Lipolytic Action of Toxohormone-L from Cancerous Ascites Fluid)

  • 이성동;황우익
    • 한국식품영양학회지
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    • 제8권2호
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    • pp.105-109
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    • 1995
  • This study was devised to observe the inhibitory effects of 3 kinds of petroleum ether extracts (percolation by petroleum ether) from Korean red ginseng, Chinese red ginseng and American white ginseng on a lipolytic action of Toxohormone-L which has been known as lipolytic and anorexigenic factors. Toxohormone-L was obtained by partial purification of the ascites fluid from mice which had been Inoculated with sarcoma-180. The yields of petroleum ether extract from Korean red ginseng, Chinese red ginseng and American white ginseng were 0.64, 0.47 and 0.58 and respectively, indicating that the yield of Korean red ginseng was the highest. In vitro, at the concentration of 2 mg /ml, the inhibition rate of lipolysis by the petroleum ether extract of Korean red ginseng, Chinese red ginseng and American white ginseng were 55.1, 50.0 and 44.9% respectively, and the total inhibitory activity per gram of ginseng material were 18, 12 and 13 unit respectively, indicating that the Korean red ginseng was the most effective in the inhibition of the lipolysis.

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고려홍삼 다당체 성분이 암독소 호르몬-L의 체지방 분해작용에 미치는 영향 (Effect of Korean Red Ginseng Polysaccharide on Lipolytic Action of Toxohormone-L from Cancerous Ascites Fluid)

  • 이성동;신유정
    • 한국식품영양학회지
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    • 제8권2호
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    • pp.110-115
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    • 1995
  • This study was devised to observe the inhibitory effects of ethanol treatment precipitate (crude acidic polysaccharide) from Korean red ginseng on a lipolytic action of Toxohormone-L which has been known as lipolytic and anorexigenic factors. Toxohormone-L was obtained by partial purification of the ascites fluid from mice which had been inoculated with sarcoma-180. The yields of crude acidic polysaccharide from Korean red ginseng was 63.5%. In vitro, at the concentration of 500rg /ml, the inhibition rate of lipolysis by the crude acidic polysaccharide of Korean red ginseng was 38.8% and the total inhibitory activity per gram of ginseng material was 4,928 nit. In vivo, the red ginseng polysaccharide(40mg/ml in saline soon.) 16ul/g of body weight was injected to the sarcoma-180 bearing mice once In 3 days until death. The effects against the extension of life span was little but body weight gain of sarcoma-180 bearing mice decreased significantly by administration of Korean red ginseng polysaccharide compared to those of the control group.

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대황(Rheum undulatum L.)으로부터 항주름성 Elastase 저해제의 추출 및 특성 (Extraction and Characterization of an Anti-wrinkle Elastase Inhibitor from Rheum undulatum L.)

  • 이재원;이지수;이종수
    • 한국식품영양학회지
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    • 제21권2호
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    • pp.143-147
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    • 2008
  • To develop a new anti-wrinkle agent from medicinal plants, this study investigated the optimal conditions for extracting elastase inhibitor from Rheum undulatum L. Maximal extraction occurred by using 70% methanol at $50^{\circ}C$ for 24 hr; the elastase inhibitory activity was 60.4%($IC_{50}:6.7{\times}10^3{\mu}g/m{\ell}$). Systematic solvent extraction, thin layer chromatography, silica gel column chromatography, Sephadex LH-20 column chromatography, and reverse-phase high performance liquid chromatography were used in the partial purification of the elastase inhibitor. The compound was soluble in dimethylsulfoxide, methanol, and ethanol, and had maximum absorption spectra at 231.5 nm and 275.5 nm.

해양 천연물에서 분리한 면역기능 조정제 렉틴 MLA의 림프구 자극분열효과 및 면역화학적 특성 (Lymphocytes Mitogenic and Immunochemical Characteristics of the Immunomodulating Lectins, MLA, from Marine Natural Products)

  • 전경희;김장환;정시련
    • 약학회지
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    • 제39권3호
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    • pp.252-259
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    • 1995
  • Isolation, purification and characterization of biophysicochemical properties of the three new lectins, MLA-I, MLA-II, MLA-III from the hemolymph of Meretrix lusoria have been reported previously. A series of immunochemical studies were investigated in this work. The three lectins were revealed as having partial identity each other by immunodiffusim and immunoelectrophoresis. These results suggest that MLA lectins are isolectins having similar biophysicochemical properties. Particularly, MLA-I proved to be a potent mitogen for murine splenic as well as human peripheral lymphocytes, and the optimum mitogenic dose were 62.5 and 1.95 $\mu\textrm{g}$/ml, respectively.

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A Novel Anticoagulant Protein from Scapharca broughtonii

  • Jung, Won-Kyo;Je, Jae-Young;Kim, Hee-Ju;Kim, Se-Kwon
    • BMB Reports
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    • 제35권2호
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    • pp.199-205
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    • 2002
  • An anticoagulant protein was purified from the edible portion of a blood ark shell, Scapharca broughtonii, by ammonium sulfate precipitation and column chromatography on DEAE-Sephadex A-50, Sephadex G-75, DEAE-Sephacel, and Biogel P-l00. In vitro assays with human plasma, the anticoagulant from 'S. broughtonii, prolonged the activated partial thromboplastin time (APTT) and inhibited the factor LX in the intrinsic pathway of the blood coagulation cascade. But, the fibrin plate assay did not show that the anticoagulant is a fibrinolytic protease. The molecular mass of the purified S. broughtonii anticoagulant was measured to be about 26.0kDa by gel filtration on a Sephadex G-75 column and SDS-PAGE under denaturing conditions. The optimum activity in the APTT assay was exhibited at pH 7.0-7.5 and $40-45^{\circ}C$ in the presence of $Ca^{2+}$.