• 제목/요약/키워드: gel fraction

검색결과 831건 처리시간 0.03초

Chlorellaellipsoidea로 부터 polyphosphate phosphohydrolase의 분리, 정제 및 성질 (Purification and properties of polyphosphate phosphohydrolase from chlorella ellipsoidea)

  • 임영복;이영록
    • 미생물학회지
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    • 제21권3호
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    • pp.135-142
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    • 1983
  • The presence of polyphosphate phosphohydrolase (PPPH) and tripolyphosphate phosphohydrolase (TPPH) in Chlorella ellipsoidea were confirmed from the cell-free extract of the algal cells and three forms of PPPH were isolated, purified, and measured Km-Vmax value and inhibitory effect by metal ions, respectively. PPPH was most active at pH7.2, whereas TPPH at pH 7.6. Both enzymes exhibited their maximum activity at $37^{\circ}C$. For the manifestation of catalytic activity, divalent, divalent metal ions are needed, and the best activator for both enzymes was $Co^{++}\;ions\;(10^{-3}M)$. These enzymes were inhibited by $Hg^{++}\;ions\;(10^{-3}M)$ considerably. PPPH from Chlorella ellipsoidea was purified by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-Sephadex A-25, and gel filtration on Sephadex G-100, and some properties of the three different fraction with PPPH activity $(PPPH_1,\;PPPH_2,\;and\;PPPH_3)$ were found, i.e, PPPH has multiple form. The Km values of $PPPH_1,\;PPPH_2,\;and\;PPPH_3$ obstained were $6.25{\times}10^{-4}M,\;10^{-4}M-4/M,\;and\;3.33{times}10^{-4}M$ and Vmax were 3.33 mM/min, 3.33 mM/min, and 2.67 mM/min, respectively. It was shown that the types of inhibition of $Hg^{++} on the activities of three forms of PPPH were competitive inhibition.

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An FMN-Containing NADH-Quinone Reductase from Streptomyces sp. (An FMN-containing NADH-quinone reductase from streptomyces sp)

  • Youn, Hong-Duk;Lee, Jin-Won;Youn, Hwan;Lee, Jeong-Kug;Hah, Yung-Chil;Kang, Sa-Ouk
    • Journal of Microbiology
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    • 제34권2호
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    • pp.206-213
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    • 1996
  • NADH-quinone reductase was purified 22-fold from the cytosolic fraction of Streptomyces sp. Imsnu-1 to apparent hemogenity, with an overall yield of 9%, by the purification procedure consisting of ammonium, sulfate precipitation and DEAE Sephacryl S-200 and DEAE 5 PW chromatographies. Thes molecular mass of the enzyme determined by gel filtration chromatography was found to be 110 kDa. SDS-PAGE revealed that the enzyme consists of two sugunits with a molecular mass of 54 kDa. The enzyme contained 1 mol of FMN per subunit as a cofactor. The $A_{272}$ A$_{457}$ ratio was 6.14 and the molar extinction coefficients were calculated to be 20, 800 and 25, 400M$^{-1}$ $cm^{-1}$ / AT 349 AND 457 nm, respectively. The N-terminal sequence of the enzyme contained the highly conserved fingerprint of ADP-binding domain. The enzyme used NADH as an electron donor and various quinones as electron acceptors. Cytochrome c was practically inactive. Air-stable flavin semiquinone was produced by the addition of NADH to the enzyme. Also, naphthosemiquinone was detected in the reaction mixture containing the enzyme.

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Isolation of Flavonoids from the Fruits of Cornus kousa Burg

  • Lee, Dae-Young;Lyu, Ha-Na;Kwak, Ho-Young;Jung, La-Koon;Lee, Youn-Hyung;Kim, Dae-Keun;Chung, In-Sik;Kim, Sung-Hoon;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • 제50권3호
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    • pp.144-147
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    • 2007
  • Dried, unripe fruits of Cornus kousa Burg. were extracted with 80% aqueous MeOH and the concentrated extracts were partitioned between EtOAc and $H_2O$. From the EtOAc fraction, four flavonoids were isolated through repeated silica gel, ODS and Sephadex LH-20 column chromatographies followed by a preparative HPLC. Based on the spectroscopic data including NMR, MS and IR, the chemical structures of the compounds were determined as kaempferol (1), astragalin (2), hyperin (3) and isoquercitrin (4). These compounds were isolated for the first time from the fruits of this plant.

Physicochemical Properties of Enzymatically Modified Maize Starch Using 4-${\alpha}$-Glucanotransferase

  • Park, Jin-Hee;Park, Kwan-Hwa;Jane, Jay-Iin
    • Food Science and Biotechnology
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    • 제16권6호
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    • pp.902-909
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    • 2007
  • Granular maize starch was treated with Thermus scotoductus 4-${\alpha}$-glucanotransferase (${\alpha}$-GTase), and its physicochemical properties were determined. The gelatinization and pasting temperatures of ${\alpha}$-GTase-modified starch were decreased by higher enzyme concentrations. ${\alpha}$-GTase treatment lowered the peak, setback, and [mal viscosity of the starch. At a higher level of enzyme treatment, the melting peak of the amylose-lipid complex was undetectable on the DSC thermogram. Also, ${\alpha}$-GTase-modified starch showed a slower retrogradation rate. The enzyme treatment changed the dynamic rheological properties of the starch, leading to decreases in its elastic (G') and viscous (G") moduli. ${\alpha}$-GTase-modified starch showed more liquid-like characteristics, whereas normal maize starch was more elastic and solid-like. Gel permeation chromatography of modified starch showed that amylose was degraded, and a low molecular-weight fraction with $M_w$ of $1.1{\times}10^5$ was produced. Branch chain-length (BCL) distribution of modified starch showed increases in BCL (DP>20), which could result from the glucans degraded from amylose molecules transferred to the branch chains of amylopectin by inter-/intra-molecular transglycosylation of ${\alpha}$-GTase. These new physicochemical functionalities of the modified starch produced by ${\alpha}$-GTase treatment are applicable to starch-based products in various industries.

고사리 열수 추출물로부터 보체계 활성화 산성 다당의 분리 및 특성 (Isolation and Characterization of Acidic Polysaccharides Activating Complement System from the Hot Water Extracts of Pteridium aquilinum var. latiusculum)

  • 오병미;권미향;나경수
    • 한국식품영양학회지
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    • 제7권3호
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    • pp.159-168
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    • 1994
  • From the hot water extract of bracken(Pteridium aquilinum var. latiusculum), a Korean win edible plant, anti-complementary acidic polysaccharides were Isolated. Crude polysaccharide fraction(HPA-1) was obtain ed by methanol reflux, ethanol precipitation, dialysis, and lyophilization. HPA-1 contained 81.80% of total sugar, 30.40% of uronic acid, and 15.60cA of protein. HPA 1 was purified consecutively by cetavlon fractionation and chromatography including ion exchange nth DEAE-Sepharose CL 6B and gel permeation with Sephadex G-100 and Sepharose CL-6B. HPA-2- IVa and HPA-Va-2 were nearly homogeneous on HPLC and had 500,000 and 560,000 daltons of molecular weights, respectively. HPA-2-Wa consisted of fucose, galacturonic acid, and glucuronic acid at the molar ratio of 1.40 : 0.97 : 1.88. HPA-2-Va 2 was composed of rhamnose, galactose, and galacturonic acid at the molar ratio of 1.00 : 1.38 : 1.39. The polysaccharides were found to activate the C3 component both In the presence and In the absence of Ca2+ through the crossed-immunoelectrophoresis suggesting that those Involved in both classical and alternative complement pathway.

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도라지(Platycodon grandiflorum DC) 추출 성분의 암세포 증식 억제효과 (Effect of Platycodon grandiflorum DC Extract on the Growth of Cancer Cell Lines)

  • 이지영;황우익;임승택
    • 한국식품과학회지
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    • 제30권1호
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    • pp.13-21
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    • 1998
  • 도라지의 물과 석유에테르 추출물의 세포 증식 억제효과를 mouse DBA/2 strain유래의 백혈병성 임파모세포인 L1210과 인체 결장 암세포인 HCT-48 및 직장암세포인 HRT-18를 대상으로 분석하였다. 수용성 추출물보다는 석유에텔 추출물이 모든 항암세포에 대하여 현저히 높은 항암효과를 나타내었다. 도라지 석유에테르 추출물을 실리카겔 컬럼크로마토그라피에서 석유에테르과 에틸에테르 혼합을(7:3)로 얻은 분획은 더욱 높은 활성을 나타냈으며 L1210에서는 0.3mg/mL, HCT-48는 1.2mg/mL의 농도에서 대조군에 비해 -99%, -55%의 증식억제효과를 보였다.

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녹용의 약효 성분에 관한 연구(VI) 녹용의 지용성 성분 및 Pantocrin이 흰쥐 척수 신경의 Aldolase 활성에 미치는 영향 (Biochemical Studies on Antler (Cervus nippon taiouanus) (VI) Comparative Study on the Effect of Lipid Soluble Fractions of Antler Sponge and Velvet Layers and Pantocrin on the Aldolase Activity in the Rat Spinal Nerves)

  • 김영근;김경자
    • 약학회지
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    • 제27권3호
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    • pp.235-243
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    • 1983
  • In the present study, in attempt was made to observe the effect of lipid components in pantocrin and antler velvet and sponge layers extracts on the aldolase activity in the rat spinal nerves. The antler chloroform-methanol (2:1) extracts are known to be composed of neutral lipids, cholesterol, cerebrosides, spingomyelin, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine and gangliosides. The antler extracts were studied on the aldolase activity in the rat spinal nerves. The aldolase activity was measured by the method of Lehninger. To investigate the components of tile extracts which affect the enzyme activity, the chloroform-methanol (2:1) extracts of antler were fractionated into petroleum ether soluble and insoluble fractions and their effects on the enzyme activity were compared. It was found that the chloroform-methanol (2:1) extracts of the antler sponge and velvet layers as well as those petroleum ether soluble fractions obtained from the chloroform-methanol (2:1) extracts, decrease the aldolase activity by 18-23%. However, pantocrin showed increasing effect on the aldolase activity by 42.6%. The petroleum ether insoluble fraction of the chloroform-methanol (2:1) -extract showed no significant increasing activity (about 13%) on the spinal nerves aldolase. The components of pantocrin ind the petroleum ether insoluble fractions were attempted to analysize by thin layer (silica gel) and gas liquid chromatography.

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산오이풀 잎의 성분 (Constituents of Sanguisorba hakusanensis Leaves)

  • 권원준;황완균;김일혁
    • 약학회지
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    • 제40권3호
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    • pp.262-272
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    • 1996
  • The constituents of Sanguisorba hakusanensis leaves (Rosaceae), of which the roots have been used as an astringent, hemostatics and antiphlogistics, were studied phytoche mically. From water fraction of the MeOH extract, gallic acid 3-O-${\beta}$-D-(6'-O-galloyl)-glucopyranoside(I), quercetin-3-O-${\beta$-D-galactopyranoside(II), quercetin-3-O-${\alpha}$-L-arabinopyranoside(III) and $2{\alpha},\;3{\beta},\;19{\alpha}$, 23-tetrahydroxyurs-12-en-28-oic acid 28-O-${\beta}$-D-glucopyranoside(IV) were isolated by column chromatographic separation using Amberlite XAD-2, ODS-gel and Sephadex LH-20. The structure of these compounds were elucidated by spectroscopic parameters of $^1H$-NMR, $^{13}C$-NMR, $^1H-^1H$ COSY, $^{13}C-^1H$ COSY, EI-Mass, FAB-Mass, IR, UV and by comparison with authentic samples.

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생쥐에서 오가피에 의한 싸이토카인 유도와 면역반응에 관한 효과 (Effects of Acanthopanacis cortex Extracts on the Cytokine-inducing and Immune response in Mice)

  • 임석린
    • 혜화의학회지
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    • 제10권2호
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    • pp.179-188
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    • 2002
  • This experimental study was carried out to evaluate the effects of Acanthopanacis cortex on Cytokine-inducing and and immune response in Mice. In order to investigate the effect of Acanthopanacis cortex, the following was performed; Cytotoxicity, in vitro, the fraction of $CD4^+$, $CD8^+$, $B220^+$ in splenic cell, gene expression of IL-12(p35), IL-12(p40), IFN-${\gamma}$, and splenic cell proliferation by Acanthopanacis cortex. Analysis of cytokine gene expression was carried out by RT-PCR amplification. Amplified PCR products were electrophoresed on 1.2% agarose gel, and the analysis (Ht) was used to 1D-density program. The results were obtained as follows. Acanthpanacis cortex showed didn't have cell toxicity under $12{\mu}g/m{\ell}$ group on mouse lung fibroblast cells. In an in vitro model using mouse peripheral blood mononuclear cells (PBMCs), extract of Acanthpanacis cortex induced multiple cytokine, including interleukin-12 (p35), interleukin-12 (p40), interferon-gamma (IFN-${\gamma}$). The extract also enhanced the percentages of the $CD4^+$, and $CD8^+$ in the untreated control were $22.1{\pm}3.3$ to $38.4{\pm}2.1$, and $5.0{\pm}0.4$ to $10.7{\pm}0.3%$, respectively. From above findings, it is suggested that Acanthopanacis cortex is able to anti-cancer and activate immune response system.

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Pot Life Assessment and Mechanical Property of Fast Curing Polyurethane Developed with Eco-friendly Pre-polymer

  • Joseph, Jessy;Moon, Junho;Kong, Tae Woong;Kim, Dong Ho;Oh, Jeong Seok
    • Elastomers and Composites
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    • 제55권1호
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    • pp.13-19
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    • 2020
  • 4,4'-Methylenebis(2-chlorobenzenamine) (MOCA)-free fast curing polyurethanes were prepared. In this study, the processibility of a fast curing polyurethane system was characterized by assessing the pot life. The obtained pot life of the polyurethane was 6-8 s, indicating that this prepolymer-curative system is appropriate for ribbon flow casting. The influence of the NCO index on the viscosity and mechanical properties was evaluated. The viscosity, tensile strength, tear strength, and hardness of the as-prepared polyurethanes showed an increasing trend, with an increase in the NCO index, whereas the elongation at break increased initially and then decreased with an increase in the NCO index. The gel fraction and crosslink density showed a direct correlation with the NCO index, which substantiated the improved mechanical properties at the higher NCO index. The coefficients of friction and abrasion deteriorated with an increase in the NCO index.