• Title/Summary/Keyword: Isolation & purification

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Role of Unstable Phenanthrene-Degrading Pseudomonas species in Natural Attenuation of Phenanthrene-Contaminated Site

  • Prakash, Om;Lal, Rup
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.79-87
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    • 2013
  • An unstable yet efficient phenanthrene-degrading bacterium strain Ph-3 was isolated from a petroleum-contaminated site at the Mathura Oil Refinery, India. The strain was identified as Pseudomonas sp. using a polyphasic approach. An analysis of the intermediates and assays of the degradative enzymes from a crude extract of phenanthrene-grown cells showed a novel and previously unreported pattern of 1, 2-dihydroxy naphthalene and salicylic acid production. While strain Ph-3 lost its phenanthrene- degrading potential during successive transfers on a rich medium, it maintained this trait in oligotrophic soil conditions under the stress of the pollutant and degraded phenanthrene efficiently in soil microcosms. Although the maintenance and in vitro study of unstable phenotypes are difficult and such strains are often missed during isolation, purification, and screening, these bacteria constitute a substantial fraction of the microbial community at contaminated sites and play an important role in pollutant degradation during biostimulation or monitored natural attenuation.

Isolation of a Pseudomonas aeruginosa Strain Producing an Extracellular Alkaline Lipase Catabolitely Regulated by Glucose, and Purification of the Lipase (포도당에 의해 합성억제되는 알카리성 Lipase를 생성분비하는 Pseudomonas aeruginosa 균주의 분리 및 이 효소의 정제)

  • 이정미;김란숙;김병오;박영덕;진익렬
    • Microbiology and Biotechnology Letters
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    • v.21 no.3
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    • pp.239-246
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    • 1993
  • Producing an extracellular alkaline lipase, this isolate JM123 was identified as a Pseudomonas aeruginosa strain from the results of the analyses of its morphological, biochemical and physiological properties. This strain showed the highest productivity of alkaline lipase when grown at pH 9.0 and 30C for 13-20 hours in the medium of 2% starch, 1% soytone, 0.5% peptone and 1% MgSO4.7H2O. However, this enzyme was greatly repressed when grown in the glucose containing medium. The culture broth was fractionated by the order of the ammonium sulfate precipitation, Sephadex G-200 gel filtration, DEAE-cellulose column chromatography, and Sephadex G-150 gel filtration.

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Triterpenoid Components of Betula latifolia $K_{OMAROV}$ -Isolation and Characterization of Triterpenoids- (자작나무 엽(葉)의 Triterpenoid 성분연구(成分硏究) -Triterpenoid의 분리(分離) 및 동정(同定)-)

  • Han, Byung-Hoon;Chi, Hyung-Joon;Han, Young-Nam
    • Korean Journal of Pharmacognosy
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    • v.4 no.4
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    • pp.167-172
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    • 1973
  • Five crystalline substances, which are positive to$L_{IEBERMAN}-B_{URCHARD}$ reaction, were isolated from the unsaponifiable fraction of the fresh leaves of Betula latifolia $K_{OMAROV}$ (Betulaceae)by silica gel column chromatographic purification. Compound $A\;(C_{29}H_{50}O,\;mp\;136^{\circ}), \;B\;(C_{30}H_{52}O_3,\;mp\;165^{\circ}), \;C\;(C_{30}H_{52}O_4,\;mp\;237^{\circ}), \;D\;(C_{30}H_{52}O_3,\;mp\;196^{\circ})\;and\; E\;(C_{30}H_{52}O_4,\;mp\;121^{\circ})$ were isolated. Compound B was characterized as a new tetracyclic triterpenoid. Compounds A, C and D were identified as ${\beta}-sitosterol$, betulatriterpene C, and betulafolienetriol, respectively.

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Isolation and purification of Cd-binding high molecular weight protein in rat liver

  • Kim, Bong-Hee;Chun, Ki-Jung
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.126-126
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    • 1995
  • 본 연구는 중금속에 의한 생체내 독성기전을 연구하고자 흰쥐간의 상층액에서 카드뮴과 잘 결합하는 HMWP들을 분리, 정재하고 나아가 생화학적 특성을 밝히고 이단백질이 카드뮴에 의해 생성되는 것인지 아니면 간의 기존 단백질인지를 밝히고자 함을 연구의 목적으로 하였다. CdCl %2 (3mg/kg body wt.)을 3일간 ip injection시킨 후 흰쥐의 간을 적출하고 균질화하여 원심분리한 crude extract를 직접 Sephacryl S-100에 흡착시켜 10mM phosphate buffer(pH 7.0)으로 용출시켰다. 용출된 fraction tube를 UV spectrometer로 흡광도를 측정하고 atomic absorption spectrophotometer로 카드뮴량을 측정하여 카드뮴량이 높은 분획을 모아서 ion exchange column chromatography(DEAE-Sepharose)에 흡착시켜 염농도 구배로 chromatography를 실시하고, 음이온 교환수지에서 흡착되지 않고 용출된 분획을 ultrafiltration으로 농축시킨 후 S-Sepharose에 흡착시켜 염농도 구배로 chromatography를 실시한 결과 두 종류의 카드뮴 결합 단백질(Cd-BP)를 분리, 정제하였다.

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Adding Effect of TCA-Precipitated Blood Powder to the Phenol Formaldehyde Resin for Plywood (합판용 페놀수지 접착제에 대한 TCA침전 혈액분말의 첨가효과)

  • Lee, Hwa-Hyoung;Lee, Jong-Shin;Jang, Sang-Sik
    • Journal of the Korean Wood Science and Technology
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    • v.24 no.2
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    • pp.15-19
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    • 1996
  • To utilize the waste materials and develope wood adhesive from isolated bloods of slaughtered cow and pig and also to prevent water pollution, simple and rapid method of isolation and purification of plasma proteins from pig bloods with trichloroacetic acid(TCA) treatment was developed. Adding of TCA-precipitated blood powder to the phenol formaldehyde resin(PF) improved dry and wet strength of plywood and resulted in fast hot pressing times.

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Isolation and Purification of Acidic Polysaccharide of Korean Red Ginseng Acting against Toxic Action of Toxohormone-L from Cancerous Ascites Fluid (암독소(Toxohormone-L)의 작용을 저해하는 홍삼산성다당체의 분리 및 정제)

  • Lee, Seong-Dong;Hwang, Yun-Gyeong
    • The Korean Journal of Food And Nutrition
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    • v.3 no.2
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    • pp.133-140
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    • 1990
  • Toxohormone-L is a lipolytic factor, found in ascites fluid of sarcoma 180-bearing mice and of patients with hepatoma A substance that inhibited the lipolytic action of Toxohormoue-L was isolated and purified from Korean red ginseng powder. This substance had a pectin-like $\alpha$-1,4-polygalacturonan backbone with some acetoxyl groups, and so was an acidic polysaccharide It inhibited Toxohormone-L Induced liploysis in a dose dependent manner at concentrations higher than 10 Ug/ml. Purified acidic Polysaccharide yield(PG4-3 and PG4-4 fraction) was about 0.03%. And also pectic acid that inhibited the lipolytic action of Toxohormone-L.

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Isolation, purification and characterization of phytohemagglutinating proteins from Korean natural products

  • Chung, See-Ryun;Jeune-Chung, Kyung-Hee;Kim, Kyong-Ae
    • Archives of Pharmacal Research
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    • v.3 no.1
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    • pp.31-36
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    • 1980
  • Seeds or beans of 55 plants belonging to 31 families were screened by using several different types of red blood cells to find new lectins. In this paper, white kidney been (phaseolus vulgaris C.) was chosen to study biochemical properties of hemagglutinating proteins(lectins). An anion exchanger, DEAE Sephadex A-50, and polyacrylamide disc gel electrophoresis were main techniques used. From three main fractions eluted by stepwise NaCl gradient in 25mM Tris-HCI buffer on DEAE Sephadex column, principal lectin was identified.

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Isolation of the Arabidopsis Phosphoproteome Using a Biotin-tagging Approach

  • Kwon, Sun Jae;Choi, Eun Young;Seo, Jong Bok;Park, Ohkmae K.
    • Molecules and Cells
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    • v.24 no.2
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    • pp.268-275
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    • 2007
  • Protein phosphorylation plays a key role in signal transduction in cells. Since phosphoproteins are present in low abundance, enrichment methods are required for their purification and analysis. Chemical derivatization strategies have been devised for enriching phosphoproteins and phosphopeptides. In this report, we employed a strategy that replaces the phosphate moieties on serine and threonine residues with a biotin-containing tag via a series of chemical reactions. Ribulose 1,5-bisphosphate carboxylase/oxygenase (RUBISCO)-depleted protein extracts prepared from Arabidopsis seedlings were chemically modified for 'biotin-tagging'. The biotinylated (previously phosphorylated) proteins were then selectively isolated by avidin-biotin affinity chromatography, followed by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). This led to the identification of 31 protein spots, representing 18 different proteins, which are implicated in a variety of cellular processes. Despite its current technical limitations, with further improvements in tools and techniques this strategy may be developed into a useful approach.

Isolation of Lipoxyeenase Isozymes from Soybean Seeds (대두 Lipoxygenase 이성효소의 분리)

  • Song, Young-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.17 no.3
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    • pp.185-190
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    • 1988
  • Soybean lipoxyeenase isozymes were isolated from acetone-defatted soybean seeds(Glycine max [L.] Merr. variety AmSoy) by ammonium sulfate fractionation, eel filtration, and ion exchange chromatoeraphy. The final preparation of lipoxygenase-1 and -2 obtained was 19- and 32-fold purified, respectively, to the crude extract. But a considerable loss of total enzyme activity occurred during purification. On 7% polyacrylamide gel electrophosis at pH 9.0, employing lipoxigenase specific staining technique, lipoxyeenase-1, -2, and -3 showed distinctive Rf values of 0.38, 0.29, and 0.33, respectively.

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