• 제목/요약/키워드: cation-exchange chromatography

검색결과 127건 처리시간 0.029초

Generation of ovine recombinant prion protein (25-232): Characterisation via anti-PrP monoclonal antibodies and CD spectroscopy

  • Yang, Su-Jeong;Thackray, Alana;Bujdoso, Raymond
    • 한국동물위생학회지
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    • 제28권4호
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    • pp.393-405
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    • 2005
  • In prion pathogenesis, the structural conversion of the cellular prion protein $(PrP^c)$ to its abnormal isomer $(PrP^{Sc})$ is believed to be a major event. The susceptibility or resistance to natural sheep scrapie is associated with polymorphisms of host PrP gene (PRNP) at amino acid residues 136, to a lesser extent 154. The 112 residue in ovine PrP displays a natural polymorphism, Methionine to Threonine, which has not been thoroughly investigated. However the cell-free conversion assay showed that ARQ with Thr112 $(T_{112}ARQ)^{1)}$ presents lower convertibility to $PrP^{Sc}$than wild type ARQ $(M_{112}ARQ)$ [1] In this study we generated ovine recombinant PrPs of 112 allelic variants by metal chelate affinity chromatography and cation exchange chromatography. The final purity of the ovine PrP ARQ was more than $95\%$. These variants showed similar immunoreactivity against anti-PrP monoclonal antibodies in Western blot and ELISA. The refolded $M_{112}ARQ$ and $M_{112}ARQ$ presented the secondary structural content to similar extent via CD spectroscopy analysis. The inherited structural features of $M_{112}ARQ$ and $M_{112}ARQ$ under the different biophysical conditions are in the middle of investigation.

미국자리공(Phytolacca americana L.) 뿌리의 항균 펩타이드 정제 및 특성연구 (Purification and Charaterization of Antimicrobial Peptide from Roots of Pokeweed)

  • 김정주;장혜영;김재호
    • Applied Biological Chemistry
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    • 제46권4호
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    • pp.385-390
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    • 2003
  • 미국자리공(Phytolacca americana L.) 뿌리에서 항균 펩타이드를 분리하여 PAMP-r로 명명 하였다. 분리는 DEAE-cellulose, sephadex G-75, Mono S, Resource RPC등을 거쳐 이루어 졌으며 최종산물은 15% SDS-PAGE 에서 약 4,900 Da의 분자량을 보였다. PAMP-r는 광범위한 항균활성을 나타냈으며 온도와 pH변화에도 높은 안정성을 보였다. 항균활성은 $80^{\circ}C$에서 30분동안 유지될 뿐만 아니라 pH도 3.0의 산성에서부터 pH 8.0의 알카리에 이르기까지 광범위한 범위 안에서 안정하게 나타났다.

New Antimicrobial Activity from Korean Radish Seeds (Raphanus sativus L.)

  • Park, Jong-Heum;Shin, Keuyn-Kil;Hwang, Cher-Won
    • Journal of Microbiology and Biotechnology
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    • 제11권2호
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    • pp.337-341
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    • 2001
  • To isolate antifungal substances from Korean radish (Raphanus Sativus L.) seeds, various purification techniques such as DE52 cellulose anion exchange, SP-Sephadex C-25 cation exchange, and Sephadex G-50 gel filtration chromatographies were used. The molecular masses of two purified R. sativus antifungal proteins (RAPs) were estimated to be about 6.1 kDa (RAP-1) and 6.2 kDa (RAP-2) by SDS-PAGE, and 5.8 kDa(RAP-1) and 6.2 kDa (RAP-2 by a gel filtration chromatography, respectively. Purified proteins RAP-1 and 2 clearly exhibited different growth inhibitory activities against other microorganisms like Candida albicans and Saccharomyces cerevisiae. Although they have similar molecular masses, both RAP-1 and 2 proteins are not identical because their microbial inhibitory actions were different. Therefore, RAP-1 could be a new antifungal protein when compared with the antifungal activities of 2S albumins, Rs-AFPs, Mj-AMPs, chitinase, glucanase, permatin, and ribosome inactivating proteins, all of which are anifungal proteins of plants.

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한국산(韓國産) 백삼(白蔘)과 수삼(水蔘)의 Polyamine 성분에 관한 연구 (Polyamine Constituents of Korea White Ginseng and Fresh Ginseng Root)

  • 김영중;한대석;조현수;안상미;허훈;구향자
    • 생약학회지
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    • 제14권2호
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    • pp.39-43
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    • 1983
  • Polyamine constituents of fresh Panax ginseng root and dried white ginseng were extracted with 5% trichloroacetic acid, respectively. The isolation of polyamine constituents was conducted by the cation exchange chromatography using $Dowex-50W{\times}8$ resin and the detection was performed with TLC. Identification of the polyamine was carried out by the methods of IR, NMR, MS spectroscopy and GLC. Polyamine constituents of white and fresh ginseng root were identical and composed of five different polyamines. The major polyamine in white and fresh ginseng root was determined as putrescine.

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MC-50 싸이클로트론을 이용한 In-111 제법 개발 (The Production of Carrier Free Indium-111 with MC-50 Cyclotron)

  • 전권수;서용섭;양승대;채종서;지광수;이종두
    • 대한방사선기술학회지:방사선기술과학
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    • 제13권2호
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    • pp.43-49
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    • 1990
  • [ $^{111}$ ]In has wide applications in nuclear medicine for labelling and in-vivo distribution studies. A method is developed for the production of carrier free $^{111}$In using the reaction $^{nat.}Cd(p,\;xn)$ $^{111}$In with MC-50 cyclotron. Carrier free $^{111}$In was separated from the irradiated metallic cadmium by liquid-liquid extraction and cation exchange chromatography. The yield of $^{111}$In at EOB is $0.8mCi/{mu}\;Ahr$ and the nuclidic purity is over 99%. $^{111}In-DTPA\;and\;^{111}In-bleomycin$ were prepared for medical study.

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Streptoverticillum morbarense로부터 생산되는 Transglutaminase 분리 및 식품에의 적용

  • 유재수;신원선;엄태붕;김영수;정용섭
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2002년도 생물공학의 동향 (X)
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    • pp.500-503
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    • 2002
  • In order to improve the physical properties of food such as texture and food self-life. Transglutaminase(mTG) from Streptoverticillum morbarense was prepared. In the preliminary experiments, presence of proteases in the crude enzyme did not improve the texture of dough, which mean the inteference of mTG reaction by the proteases. Among the cation exchange resins tested for the removal of proteases, Monoplus S 100(Bayer, Germany) was the most efficient resin with 20 fold increase in the mTG/protease activity ratio. By further purification steps with a quaternary ammonia salt resin and a gel permeation chromatography, proteases were effectively removed from the preparation. Therefore, the improvement of flour texture was shown by adding the protease-free mTG.

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대두가수분해물로부터 새로운 항혈전성 펩타이드, SSGE와 DEE의 분리

  • 이경애;김승호
    • 식품기술
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    • 제17권3호
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    • pp.69-74
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    • 2004
  • A soy protein hydrolysate was found to inhibit rat platelet aggregation induced by ADP, an aggregating agent. To find out its principal antiplatelet peptide(s), the soy protein hydrolysate was separated successively by gel filtration chromatography, revere-phase HPLC, and cation exchange HPLC. During the course of separation, we observed that most fractions had antiplatelet effects, which suggests that most peptides have some degree of antiplatelet effect. Following the inhibitory fractions, we purified and identified two new peptides, SSGE and DEE, by LC-electrospray ionization MS and peptide equencing. Both peptides were highly hydrophilic. The concentrations to obtain 50% inhibition ($IC_50$) of the aggregation intensity were approximately $\458muM$ and $\485muM$, respectively, for SSGE and DEE.

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Bacillus thurinRiens영의 주입에 의해 유도된 담배나방의 항세균 단백질의 정제 (Purification of Antibacterial Proteins in Heliothis assulta Injected with Bacillus thuringiensis)

  • 유종명;정성은이형철
    • 한국동물학회지
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    • 제37권2호
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    • pp.274-280
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    • 1994
  • 담배나방(Heliothis assulta) 혈림프 항세균성 단백질의 특성을 알아보기 위해 종령 유충의 혈강으로 고라구연us thuringiens논 균을 주입하였다. 혈림프의 항세근적 활성은 주입후 48시간 경과군에서 가장 높았으며, E. coli와 풍 thuringiensis에 대하여 항세균성을 보이는 두 개의 단백질(G-1 및 G-11)들을 cation exchange chromatography와 gel filtration을 통해 정제하였다 49KD와 46KB의 두 subunit로 구성된 G-1은 높은 glvcine(17 13 mole %) 및 alanine(13.88 mole %) 함량을 보였으며, 14KD의 단일 pclvpeptide인 G-11에서는 histidine(16 14 mole %)과 glvclne(14.27 mole %)의 함량이 높아, 이러한 높은 glvcine 함량에 있어서 곤충에서 나타나는 다른 항세균성 단백질과 공통점을 보인다.

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Isolation of Polypeptide Fractions from Different Park offered Ginseng

  • Larina, Ludmila;Muranova, T.;Cho, B.G.;Park, H.
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1998년도 Advances in Ginseng Research - Proceedings of the 7th International Symposium on Ginseng -
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    • pp.160-167
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    • 1998
  • Chemical studies of nitrogen compounds of Panax ginseng seem relatively rare, Probably due to the isolation difficulties, subsequently the investigations of biological activities are little. The experimental conditions were established for highly complete extraction of peptides (basic, acidic and neutral) from Panax ginseng. This task was achieved by applying the follow isolation procedure: 1 , the sequential extraction with water, 0.1% TFA in 20% acetonitril and buffer pH 6.5 (water-pyridine-acetic acid 100:3:900) : 2, fractionation by ultrafiltration : 3, n-butanol extraction 4, cation- and anion-exchange chromatography : 5, chromato-electrophoresis. The comparison of red ginseng (xylem Sl pith part) and red ginseng inside white (xylem Sc pith part) was also provided. To analyze the peptide mixture the chromato-electrophoresis method of separation was applied. Optimal conditions for peptides mapping of sample were explored. Our experiments revealed the quantitative difference of peptide between xylem & pith and phloem & cortex part. We have also found the qualitative difference in the composition of polypeptides between normal red ginseng (xylem Sc pith part) and red ginseng inside-white (xylem St pith part)

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D-xylose에서 D-xylulose로의 전환을 위한 D-xylose Isomerase의 정전기적 고정화 (Electrostatic Immobilization of D-Xylose Isomerase to a Cation Exchanger for the Conversion of D-Xylose to D-Xylulose)

  • 항누엔티;김성건;권대혁
    • 한국미생물·생명공학회지
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    • 제40권2호
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    • pp.163-167
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    • 2012
  • D-Xylose는 Saccharomyces cerevisiae에서 기질로 이용될 수 없어 S. cerevisiae가 이용 가능한 D-xylulose로의 전환이 요구된다. 효소고정화 시스템을 통한 D-xylose로부터 D-xylulose로의 전환을 위해 대장균의 D-xylose 이성화효소(XI)의 카르복시 말단에 양이온 교환수지를 이용한 단순 정제 및 고정화가 가능하도록 10-arginine tag(R10)을 융합하였다. 융합단백질인 XIR10은 재조합 대장균에서 과잉발현되었고 단일 단계의 양이온 교환 크로마토그라피를 통하여 고순도로 정제되었다. 정제된 XIR10은 카르복시 말단의 10-arginine tag과의 정전기적 상호작용을 통하여 양이온 교환수지에 고정화되었다. 고정화 및 비고정화된 XIR10은 넓은 범위의 pH 및 온도에서 비슷한 D-xylose 이성화효소 활성을 보였다. 이 결과는 양이온 교환수지로의 고정화는 XIR10의 효소적 기능에 영향을 주지 않는다는 것을 나타낸다. 고정화된 XIR10의 최적화된 조건에서 D-xylose의 25%는 D-xylulose로 이성화되었다. 본 연구의 결과들은 10-arginine tag과 양이온 교환수지간의 상호작용을 통해 정전기적 고정화된 XIR10이 D-xylose로부터 D-xylulose로의 전환에 응용 가능하다는 것을 명확하게 보여주었다.