• Title/Summary/Keyword: Ion gel

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Purification and Characteristics of ${\beta}$-Galactosidase from Strawberry (딸기 ${\beta}$-Galactosidase의 정제 및 생화학적 특성)

  • Lee, Kwang-Hee;Yoon, Kyung-Young;Kim, Kwang-Soo;Kim, Nam-Woo;Shin, Seung-Ryeul
    • Food Science and Preservation
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    • v.7 no.2
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    • pp.201-206
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    • 2000
  • ${\beta}$-Galactosidase was extracted and purified from strawberry. The purified ${\beta}$-Galactosidase from strawberry was investigated their physicochemical characteristics. ${\beta}$-Galactosidase was purified 25.74 fold from strawberry. The purification procedure include ammonium sulfate fraction, acetone powder treatment and gel and ion exchange chromatography. Yield of the enzyme purification was 18.11%. The purified enzyme has native molecular weight of 116,000 dalton. Vmax value and Km value of ${\beta}$-Galactosidase were 0.077 mM ONPG/ml/15mim and 1.75x10-2mM, respectively. The optimum temperature and pH of ${\beta}$-Galactosidase were 43$^{\circ}$C and pH 4.0, respectively. The ${\beta}$-Galactosidase activity was stable below 50$^{\circ}$C and at pH 4.0 to pH 6.0. Among the metal ions Ca and Mg were did not affect, whereas K, Cu and Zn show a little effect on the enzyme activity. The ${\beta}$-Galactosidase activities were inhibited by treatment with EDTA and SDS.

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Purification and Properties of a Collagenolytic Protease Produced by Marine Bacterium Vibrio vulnificus CYK279H

  • Kang, Sung-Il;Jang, Young-Boo;Choi, Yeung-Joon;Kong, Jai-Yul
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.593-598
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    • 2005
  • A collagenolytic enzyme, produced by Vibrio vulnificus CYK279H, was purified by ultrafiltration, dialysis, Q-Sepharose ion exchange and Superdex-200 gel chromatography. The enzyme from the supernatant was purified 13.2 fold, with a yield of 11.4%. The molecular weight of the purified enzyme was estimated by SDS-PAGE to be approximately 35.0kDa. The N-terminal sequence of the enzyme was determined as Gly-Asp-Pro-Cys-Met-Pro-Ile-Ile-Ser-Asn. The optimum temperature and pH for the enzyme activity were $35^{\circ}C$ and 7.5, respectively. The enzyme activity was stable within the pH and temperature ranges 6.8-8.0 and $20{\sim}35^{\circ}C$, respectively. The purified enzyme was strongly activated by $Zn^{2+},\;Li^{2+},\;and\;Ca^{2+}$, but inhibited by $Cu^{2+}$. In addition, the enzyme was strongly inhibited by 1, 10-phenanthroline and EDTA. The purified enzyme was suggested to be a neutral metalloprotease.

Isolation, Purification and Characterization of Polysaccharides that induce in vitro Immuno-Stimulation of Macrophases derived from Liquid Culture of Cordyceps militaris

  • Kwon, Jeong-Seok;An, Hyo-Sil;Hong, Eock-Kee
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.287-292
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    • 2005
  • The crude polysaccharides(C-CPF, C-CPM, C-CPB) derived from fruiting body, mycelia and mycelia free broth of cordyceps militaris were obtained by ethanol precipitation of hot water extracts. After a batch fermentation of C. militaris was carried out in a 5 L jar vessel, endo-polysaccharide and exo-polysaccharide were obtained. They were demonstrated as the hetero polysaccharides which were composed of glucsose, galactose and mannose by performed with HPAEC(high pH anion exchange chromatography) and conformation of random coil by its complex forming ability with congo red reagent. They were purified by ion exchange (DEAE-cellulose) and gel filtration chromatography. They were monitered by phenol-sulfuric acid method and Bradford method. The NO induction activities of crude polysaccharides and purified polysaccharides derived from mycelia free broth were enhanced rather than LPS(lipo polysaccharide) which was used as a general NO inducer. These effects presumably contibute to the antitumor activities. The homogenieties and molecular weights of polysaccharides were determined by using Sepharose CL-6B. The yield, molecular weights and NO induction activities of C-CPFN Fr.III, C-CPMN Fr.III, C-CPBN Fr.II were 0.387, 0.408 and 0.153, 127 K 210 K and 36 K, 40.79%, 88.72%, and 104.17%, respectively.

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Isolation of Iron-Binding Peptides from Sunflower (Helianthus annuus L.) Seed Protein Hydrolysates (해바라기씨박 단백질 가수분해물로부터 철분 결합 펩타이드의 분리)

  • Choi, Dong Won;Kim, Nam Ho;Son, Kyung Bin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1162-1166
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    • 2013
  • Proteins from sunflower seeds were hydrolyzed with Alcalase and Flavourzyme to isolate iron-binding peptides. The optimal hydrolysis conditions were determined. Hydrolysates were filtered under a 3 kDa membrane and iron-binding peptides separated from the hydrolysates using ion exchange and gel permeation chromatographic methods. A fraction with the highest iron-binding activity (Fe/peptide, 0.69), F22, was obtained. These results suggest that fractions isolated from sunflower seed protein hydrolysates can be applied toward the production of iron supplements.

Development of Image-based Fluorescence Photobleaching Technique for Measuring Macromolecule Diffusion in Biological Porous Medium (생체 다공성 매질에서 분자 확산 측정을 위한 영상 기반 형광 광표백 기법 개발)

  • Lee, Dong-Hee;Lee, Jeong-Hoon;Park, Choon-Ho;Kim, Jung-Kyung
    • Journal of the Korean Society of Visualization
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    • v.7 no.1
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    • pp.9-13
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    • 2009
  • Fluorescence recovery after photobleaching (FRAP) has been widely used for the measurement of molecular diffusion in living cells and tissues. We developed an image-based FRAP (iFRAP) technique using a modified real-time microscope and a 488 nm Ar-ion laser. A fractional intensity curve was obtained from the time-lapse images of fluorescence recovery in the bleached spot to determine the diffusion coefficient of fluorescently labeled macromolecules in porous medium. We validated iFRAP through experiments with agar gels (0.5% and 1.5% w/v) containing FITC-Dextrans (10, 70 and 500 kDa MW). Further validation was performed by a Monte Carlo approach, where we simulated the three-dimensional random walk of macromolecules in agar gel model. Diffusion coefficients were deduced from the mean square displacement curves and showed good agreements with those measured by iFRAP.

Studies on the Development of a Thrombolytic Agent from Korean Snake Venom I. Purification of a Protease from the Venom of A. bromhoffi brevicaudus (한국 독사독으로부터의 혈전 용해제 개발에 관한 연구 I. 살모사(A. bromhoffi brevicaudus) 사독 Protease의 정제에 관한 연구)

  • Lee, Mun-Han;Kim, Byoung-Jae;Rim, Jong-Seop;Lee, Hang;Lee, Hye-Suk;Kim, Jong-Ho;Chai, Chang-Su
    • Biomolecules & Therapeutics
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    • v.3 no.2
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    • pp.159-164
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    • 1995
  • Fibrinolytic and fibrinogenolytic activities of the venoms from the Korean snakes, Agkistrodon caliginosus, nosus, Agkistrodon saxatilis and Agkistrodon blomhoffi brevicaudus were compared by fibrin-plate method and polyacrylamide gel electrophoresis, respectively. The venom from A. blomhoffi brevicaudus showed the highest degree of fibrin(ogen)olytic activity, and a protease with the fibrin(open)olytic activity was purified by p-amino-benzamidine affinity chromatography and DEAE ion-exchange chromatography. The purified enzyme had a molecular weight of 50,800 and a capability to degrade the B$\beta$-chain of fibrinogen preferentially to the $A\alpha$-chain, but not the ${\gamma}$-chain. Fibrinolytic activity of the purified enzyme was approximately 3.8 plasmin unit/mg protein.

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Albumin Fractions from Different Species Stimulate In Vitro Progesterone Production by Granulosa Cells in Buffalo

  • Taneja, R.;Bansal, P.;Sharma, M.K.;Singh, D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.11
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    • pp.1559-1563
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    • 2002
  • The ovarian follicular fluid was found to contain steroidogenesis stimulatory protein similar to albumin from human and buffalo. Therefore, the albumins from various species, commercial and purified, were studied for their steroidogenic effect on progesterone secretion by granulosa cells from buffalo ovaries, during culture. A dose of $20{\mu}g$ of bovine serum albumin was optimum to exhibit maximum progesterone secretion on day 6 of culture, in medium ($350{\mu}l$) containing $10^5$ cells. Among commercial albumins, chicken albumin showed highest effect on progesterone secretion, which was followed by albumins from goat, bovine, human, sheep and rat, respectively at day 6 of culture. The albumins were also purified from blood serum of buffalo, goat and rat using salt fractionation, ion-exchange chromatography, gel filtration and SDS-PAGE. The highest stimulatory effect on progesterone secretion was shown by albumin purified from buffalo blood serum and lowest by that from rat blood. Comparatively the buffalo and goat albumins were more biologically active than commercial albumins. The presence of some active molecules conjugated with freshly purified albumins may be responsible for better stimulatory effect.

Purification and Characterization of Cop, a Protein Involved in the Copy Number Control of Plasmid pE194

  • Kwak, Jin-Hwan;Kim, Jung-Ho;Kim, Mu-Yong;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.291-297
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    • 1998
  • Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.

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Preparation of Fe-AC/$TiO_2$ composites and pH dependence of their Photocatalytic activity for methylene blue

  • Meng, Ze-Da;Zhang, Kan;Oh, Won-Chun
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.19 no.5
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    • pp.268-276
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    • 2009
  • In this study, activated carbon (AC) was treated with ferric ion by a sol-gel method. The compound (Fe-AC) was employed for the preparation of Fe-activated carbon/$TiO_2$, (Fe-AC/$TiO_2$) composites. The prepared Fe-AC/$TiO_2$ composites were characterized with surface properties, structural crystallinity, elemental identification and photocatalytic activity. The SEM results showed that ferric compounds and titanium dioxide were fixed onto the AC surfaces. The XRD results showed that Fe-AC/$TiO_2$ composites mostly contained anatase phase. EDX showed the presence of C, O, and Ti with Fe peaks in all samples. Its photocatalytic degradation effect was evaluated with the degradation behavior of the methylene blue (MB) solution. MB degradation could be attributed to the synergetic effects of adsorption, photo-degradation of $TiO_2$ and photo-Fenton of Fe component. The degradation rate for this photocatalysis was evaluated as a function of the concentration of the dye, the amount of $TiO_2$ and the pH. Photocataytic activity is good at activity pH.

Crystal Chemistry and Dielectric Properties of $Bi_4Ti_3O_{12}$ by the Substitution of Rare Earth Elements (Y, Nd, Sm, Gd) (희토류원소(Y, Nd, Sm, Gd)의 치환에 의한 $Bi_4Ti_3O_{12}$의 결정화학 및 유전물성)

  • 고태경;방규석
    • Journal of the Korean Ceramic Society
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    • v.32 no.10
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    • pp.1178-1188
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    • 1995
  • Bi4Ti3O12 (BIT) and its rare earth (Y, Nd, Sm, Gd)-substituted derivatives were synthesized using a sol-gel method to investigate their microstructures, cystal structures and electrical properties depending on the subsituted elemetns. Nd- or Sm-substitution into BIT appeared to be favorable, while Y- or Gd-substitution occurred with a pyrochlore phase. This suggests that a smaller trivalent rare earth ion may not be favorable in the structure of BIT. The rare earth derivatives showed that their particle sizes and shapes were considerably different depending on the kinds of substituted elements. Y-substitution resulted in developing a relatively even particle size and a dense microstructure. In structure, they may be similar to the pseudo-orthorhombic BIT but close to a paraelectric tetragonal phase. Their a (or b) axes were shortened, compared to the one of BIT. Such a distortion may result a decrease in the tilting of TiO6. BIT and the derivatives showed that their dielectric constants and losses were 40~120 and less than 0.03, respectively in the frequency range of 1~10 MHz. The dielectric loss of Y-substituted derivative was the lowest one and changed a little to frequency. Curie points were observed in all the derivatives like BIT to suggest that they would be ferroelectric. The temperature stability of the delectric properties of the derivatives below the Curie points were relatively better than the one of BIT.

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