• Title/Summary/Keyword: Anion Exchange

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Kinetic and Chemical Mechanism of Pig Liver Methenyltetrahydrofolate Synthetase (돼지 간 유래의 methenyltetrahydrofolate synthetase 반응메커니즘)

  • Cho, Yong-Kweon
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1036-1041
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    • 2008
  • Methenyltetrahydrofolate synthetase extract was obtained from mouse liver and purified via $30{\sim}70%$ ammonium sulfate fractionation, Fast Q anion exchange and phenyl agarose chromatography. HPLC gel chromatography and SDS-polyacrylamide electrophoresis experiments showed that the enzyme is a monomer with molecular weight of 23 kDa. Optimum temperature and pH were $35^{\circ}C$ and 6.5, respectively. The enzyme was chemically modified only by tetranitromethane and 1-ethyl-3- (3-dimethyl aminopropyl)-carbodiimide (EDC), indicating that tyrosine and carboxylate are in the active site. pH studies showed that 2 tyrosines are involved in the binding of the substrates and a carboxylate in catalysis. Therefore, the chemical mechanism of the enzyme is likely that 2 tyrosines bind to ATP and 5-formylTHFand a carboxylate acts as a general base.

Development of recombinant human chorionic gonadotropin (hCG) using high-density culture technique of suspension-adapted chinese hamster ovary (CHO) cells

  • Na, Kyu-Heum;Kim, Seung-Chul;Seo, Kwang-Seok;Lee, Sung-Hee;Kang, Soo-Hyung
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.37-37
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    • 2005
  • Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone consisting of non-covalently linked two subunits, the ${\alpha}$ and ${\beta}$ subunit. It has been used as a infertility drug for ovulation to mimic luteinizing hormone $(LH).^{1)}$ A stable cell line was established by transfection of Rc/CMV-i-dhfr-hCG, expression vector containing hCG ${\alpha}-$ and ${\beta}-genes$, into dihydrofolate reductase-deficient CHO cells and subesquent methotrexate-mediated gene amplification. Anchorage-dependent CHO cells were adapted into a serum-free and/or animal component-free suspension medium through gradual serum weaning for the hCG production. The established cell line showed typical morphological characteristics and growth profile of CHO cells, and could produce FSH with passage-to-passage consistency. The high density perfusion culture of the CHO cells was carried out in Celligen Plus bioreactor equipped with a spin-filter as a internal cell retention device. The cell density reached up to $>1x10^{7}$ cells/ml in less than 7 days and a perfusion-control strategy based on cellular consumption rates of glucose was $established.^{2)}$ Biologically active recombinant hCG was purified by a series of chromatographic steps including anion exchange chromatography and hydrophobic interaction chromatography to homogeneity. The highly purified recombinant hCG was characterized for physicochemical, immunological and biological properties.

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Determination of $^{241}$Am and $^{241}$Cm in Radwaste Samples (방사성폐기물 시료 중 $\^{241}$Am과 $\^{244}$Cm의 정량)

  • Joe Kih Soo;Kim Tae Hyun;Jeon Young Shin;Jee Kwsng Yong;Kim Won Ho
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.3 no.1
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    • pp.1-7
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    • 2005
  • Anion exchange chromatography and HDEHP extraction chromatography using DTPA-lactic acid as an eluent were applied in series for the separation of $^{241}$Am and $^{244}$Cm in radwaste samples. The separated elements were determined by electrodeposition at the sodium hydrogen sulfate-sodium sulfate buffer solution followed by alpha-spectrometry. The recovery yields of $^{241}$Am and $^{244}$Cm were 85.2$\pm$$15.3\%$, respectively, from the synthetic solution of spent nuclear fuel sample. The amounts of 241Am and 2440m determined in radwaste sample solutions of condensate bottoms were at the range of 1.5-1.9 Bq/g and -1.7 Bq/g, respectively.

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Purification and Chemical Properties of Anti-complementary Polysaccharide from Capsici Fructus (고추의 항보체 다당의 정제와 특성)

  • Ra, Kyung-Soo;Yamada, Haruki;Sung, Ha-Jin;Cyong, Jong-Chol;Yang, Han-Chul
    • Applied Biological Chemistry
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    • v.32 no.4
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    • pp.378-385
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    • 1989
  • Water-soluble crude polysaccharide (CAP-0) obtained from the Capsici Fructus(the fruits of Capsium annuum L.) showed a potent anti-complementary activity. The anti-complementary activity did not change by pronase digestion of CAP-0, but decreased by the periodate oxidation. CAP-0 was fractionated into four polysaccharide fractions, CAP-1,2,3 and 4, by the addition of cetyltrimethylammonium bromide. CAP-1, showing the highest anti-complementary activity, was refractioned by anion-exchange chromatography to give three major fractions(CAP-1-III, IV and V). CAP-1-III was shown to have low anti-complementary activity, but CAP-1-IV and V had high activity. CAP-1-III and IV were purified on Sephadex G-100 to give each two fractions(CAP-1-IIIa and IIIb, CAP-1-IVa and IVb), respectively. From the results of gel filtration and electrophoresis, these four fractions and CAP-1-V were found to be homogeneous polysaccharides. High molecular polysaccharies(M.W. CAP-1-IIIa 70,000, IVa 195,000, V 140,000) showed relatively higher anti-complementary activity than low molecular polysaccharides(CAP-1-IIIb and IVb).

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Candida rugosa Lipase-Catalyzed Production of Optically Pure S-(+)-Ketoprofen (Candida rugosa 리파제를 이용한 광학적으로 순수한 S-(+)-Ketoprofen의 생산)

  • 김민곤;최순자;최원아;김철호;정봉현
    • KSBB Journal
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    • v.14 no.2
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    • pp.225-229
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    • 1999
  • Enzymatic resolution reactions were investigated using Candida rugosa lipase for the production of potically pure S-(+)-Ketoprofen. When the enzymatic hydroysis (and esterification) of recemic ketoprofen esters (and recemic ketoprofen with alcohol) was investigated comparatively, aqueous media was more specific for S-enantiomer than organic media. In the enzymatic hydrolysis of racemic ketoprofen ethyl ester in aqueous media, optimal temperature and pH for enantioselectivity were $37^{\circ}C$ and 4, respectively. The stereoselectivity of the enzyme was enhanced by adding dialcohols such as ethylene glycol and propylene glycol. The enantiomeric ratio obtained in the 40 %(v/v) ethylene glycol was 2-fold higher than that without the additive. By adding $CH_2Cl_2$, $CHCl_3$ and $CCl_4$ (5%,v/v), the enantioselectivity was reversed. A dramatic increase in the stereoselectivity was achieved using lipase purified by anion exchange chromatography. The type A lipase(the first eluted lipase fraction) showed an enantiomeric ratio of >100, whereas the type B lipase(the second eluted lipase fraction) exhibited enantimomer ratio of 9.0 in the hydrolysis of racemic ketoprofen ethyl ester.

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Adsorption Characteristics of Cu(II) in the Presence of Surfactants on Natural Zeolites Treated Chemically and Thermally (가열 및 화학처리된 천연 제올라이트의 계면활성제 수용액 내에서 Cu(II)흡착특성)

  • Sun, Y.S.;Kim, P.K.
    • Applied Chemistry for Engineering
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    • v.7 no.5
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    • pp.849-860
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    • 1996
  • Natural zeolites which were known to occur in sedimentary clinoptilolite, were treated by thermal and chemical methods, and their adsorption characteristics of Cu(II) were studied. Analyses by FT-IR, S. E. M, TGA/DSC, and XRD showed that the adsorption capacity of Cu(II) on the zeolite decreased gradually at the temperatures of above $400^{\circ}C$. The zeolites, which treated with NaOH and NaCl, show higher absorbility in acid solution. The adsorption rate of Cu(II) in the presence of surfactants(LAS, POE AE) on the thermally or chemically treated zeolites varied depending on pH, anion or nonion surfactants.

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Immunomodulating Effect of the Lectin from Allomyrina dichotoma (장수풍뎅이(Allomyrina dichotoma) 렉틴의 면역기능 증강효과)

  • Jeune, Kyung-Hee;Jung, Mi-Yeun;Choi, Soo-Jeong;Lee, Jong-Wook;Park, Won-Hark;Cho, Se-Hoon;Lee, Seung-Ho;Chung, See-Ryun
    • Korean Journal of Pharmacognosy
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    • v.32 no.1 s.124
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    • pp.31-38
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    • 2001
  • A lectin was purified from Allomyrina dichotoma (ADL) by physiological saline extraction, ammonium sulfate fractionation, anion exchange column chromatography on DEAE Sephadex A-50 and gel filtration column chromatography on Sephadex G-200. Several biochemical properties of ADL were characterized as follows: ADL from gel filtration column chromatography showed single band on SDS-PAGE. ADL agglutinated the erythrocytes of rabbit and human A, B, O, AB. Agglutinability was relatively stable at basic pH, and was stable at temperature below $40^{\circ}C$. Agglutinability was not affected by metal ions and EDTA. This lectin was proved to be a glycoprotein which contains 0.47% of sugars. The molecular weight of ADL was estimated to be 97,000 dalton by SDS-PAGE. By amino acid analysis, ADL exhibited high amounts of aspartic acid. The lectin's immunomodulating effect was measured as cytokine production. The productions of 5 cytokines $(IL-1{\alpha},\;IL-2,\;IL-6,\;IFN{\gamma}\;and\;TNF{\alpha})$ from peripheral blood mononuclear cells were measured by ELISA. The lectin induced the highest secretion of IL-2 at 8 hr, $TNF{\alpha}$ at 4 hr, and $IFN{\gamma}$ at 24hr, respectively. These results suggest that ADL can elicit the production of detectable cytokines from PBMC.

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Development of cathode catalyst layer using non-Pt catalyst for SAFC (비백금 촉매를 이용한 알칼리 연료전지 캐소드 전극 개발)

  • Park, SeokHee;Choi, YoungWoo;Yim, SungDae;Kim, ChangSoo;Park, Seung Bin
    • 한국신재생에너지학회:학술대회논문집
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    • 2011.05a
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    • pp.97.1-97.1
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    • 2011
  • 저온에서 양이온 고분자막을 사용하는 고분자 연료전지의 경우 뛰어난 성능과 다양한 응용분야로 인해 많은 연구와 실증이 이루어지고 있지만 공기극에서의 느린 산소 환원반응으로 인해 백금과 같은 귀금속의 사용이 불가피하고 백금의 제한된 매장량과 높은 가격으로 인해 상용화가 늦어지고 있다. 그래서 많은 연구자들이 합금 촉매 또는 비귀금속 촉매를 이용한 전극 개발에 집중하고 있다. 알칼리 분위기에서 저가의 전이 금속들이 백금과 비슷한 활성을 보이고 고체 음이온 교환막이 개발됨에 따라 최근 알칼리 연료전지가 다시금 큰 주목을 받고 있다. 그러나 고분자 연료전지와는 달리 아직 촉매나 전해질막, 이오노머의 특성 및 메커니즘에 관해 별로 알려진 것이 없다. 본 연구에서는 직접 개발한 세공충진막 형태의 탄화수소계의 음이온 교환막과 비귀금속 공기극 촉매를 이용하여 막전극접합체(MEA)를 개발하였고 촉매 및 이오노머 함량과 같은 전극 조성, 막전극접합체의 제조 및 체결, 가습이나 가스조성 등의 단위전지 운전조건과 같은 다양한 변수에 대해에 최적 조건을 도출하고자 하였다. 공기극 촉매는 Cu-Fe/C를 이용한 상용 촉매를 이용하였고 이오노머의 경우는 탄화수소계의 상용 제품을 사용하였으며 음이온 교환막에 전극층을 형성하기 위해서는 스프레이 공정을 이용하였다. 단위전지를 통해 성능을 확인하였고 임피던스 및 CV를 통해 전기화학적인 특성을 규명하였다. 조건의 최적화를 통해 상당한 성능 향상을 이루었으나 추가적인 성능 향상 및 내구성 확보 등에 대해 계속적인 실험을 진행할 예정이다.

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Purification and Properties of Wheat Fructan Exohydrolase (밀 Frucdtan Exohydrolase의 분리 및 특성)

  • Byeong Ryong, Jeong;Thomas L, Housley
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.4
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    • pp.456-464
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    • 1996
  • Fructans are the major vegetative storage carbohydrate in wheat(Triticum aestivum L.). The depolymerization of fructans occurs by the sequential removal of terminal fructosyl residues by a specific fructan exohydrolase(FEH). The objective of this study was to isolate and characterize this enzyme in wheat. From stems and sheaths of field-grown wheat(cv. Clark), FEH was purified 356-fold using salt precipitation and a series of chromatographic procedures including size exclusion, anion exchange, and affinity chromatography. FEH had a molecular weight of 63.7 kD and an optima at pH 5.5 and 3$0^{\circ}C$. The $K_{m}$s for $\beta$(2 longrightarrow1) linked oligofructans varied, from 10 to 37mM, with the lowest $K_{m}$ for tetrasaccharide. The $V_{max}$ increased as degree of polymerization (DP) increased. Wheat FEH hydrolyzed only, $\beta$(2 longrightarrow1) linked fructans but not, $\beta$(2 longrightarrow6) linked timothy fructan or sucrose. The role of this FEH in fructan metabolism in wheat is discussed.sed.

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Effect of Amylose Content on Corn Starch Modification by Thermus aquaticus 4-$\alpha$-Glucanotransferase

  • Cho, Kyoung-Hee;Auh, Joong-Hyuck;Kim, Jung-Hwan;Ryu, Je-Hoon;Park, Kwan-Hwa;Park, Cheon-Seok;Yoo, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1201-1205
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    • 2009
  • Corn starches with different amylose contents were enzymatically modified using Thermus aquaticus 4-$\alpha$-glucanotransferase ($TA{\alpha}GTase$). Upon the enzyme treatment, the chain-length distributions of isoamylolytically debranched products became broader [degree of polymerization (DP): 3-40] than those of untreated corn starches. In addition, a variety of cycloamyloses (CAs) with different sizes were formed by the glucanotransfer activity of $TA{\alpha}GTase$. CAs with DP 5-40 were detectable in all of the $TA{\alpha}GTase$-treated corn starches. From the results of high-performance anion-exchange chromatography and high-performance size-exclusion chromatography analyses, it was suggested that the amount of CAs produced by the enzyme treatment increased as the amylose content of the starches increased. Thus, we concluded that the extent of modification of starch molecules was enhanced in proportion to amylose content by the transfer activity of $TA{\alpha}GTase$. This finding could be useful for developing an efficient process of CA production using this enzyme.