• 제목/요약/키워드: affinity-chromatograpy

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Mannobiose-Sepharose 담체합성 및 Affinity column chromatograpy에 의한 Debaryomyces sp. 유래 ${\alpha}$-Galactosidase의 정제 및 기질 특이성 (Purification and Substrate Specificity of Debaryomyces sp. ${\alpha}$-Galactosidase by Mannobiose-Sepharose Affinity Column Chromatograpy)

  • 박귀근
    • Applied Biological Chemistry
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    • 제49권3호
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    • pp.180-185
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    • 2006
  • 본 연구는 Debaryomyces sp.가 생산하는 ${\alpha}-galactosidase$의 mannobiose-sepharose 담체합성법에 의한 affinity column chromatography를 수행하여 효소정제, 효소 화학적 성질 및 galactosyl mannooligosacchrides에 대한 기질특이성 규명을 주요 목적으로 하였다. 배양시간별 활성과 pH의 변화에서 배양시간 40시간부터 활성이 증가하고 있으며, 70시간에서 25.8unit/ml의 최대활성을 나타내고 있으며, 배양초기 pH 6.0에서 시작되어 배양말기에는 pH 8을 나타내는 pH의 변화를 보였다. 최적 pH는 4.0, 최적온도는 $60^{\circ}C$이며 $pH\;3{\sim}4.5$에서 100%의 잔존활성을 나타낸 반면 pH 8.0에서는 20%로 급격히 감소하였고. 온도안정성에서 $30{\sim}50^{\circ}C$에서는 100%의 잔존활성을 나타내었으나 $70^{\circ}C$ 이상에서는 20% 이하의 잔존활성을 나타내었다. $Hg^{2+}$에 의해서 54%, $Ag^{2+}$에 의해서는 85%로 저해되었으며, 그 이외의 이온에 대해서는 큰 영향을 받지 않았다. Debaryomyces sp.유래 ${\alpha}-galactosidase$는 24시간 반응 후 melibiose, $Gal^3Man_3$를 완전 가수분해 하여 각각 galactose와 glucose, galactose와 mannotriose로 분해되었으나 $Gal^3Man_4$에 대해서는 12시간 반응시켜도 기질로부터 galactose 유리가 불가능함이 확인되었다.

Sucrose-permeability Induced by Reconstituted Connexin32 in Liposomes.

  • Rhee, Senng-Keun;Hong, Eun-Jnng
    • BMB Reports
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    • 제28권2호
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    • pp.184-190
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    • 1995
  • Functional study of the gap junction channel has been hindered by its inaccessibility in situ. Identification of forms of this channel in artificial membrane has been elusive because of the lack of identifying channel physiology. Connexin32 forms gap junction channels between neighboring cells in rat liver. Connexin32 was affinity-purified using a monoclonal antibody and reconstituted into artificial phospholipid vesicles. The reconstituted connexin32 formed channels through the vesicle membrane that were permeable to sucrose (Stokes radius: $5{\AA}$). The permeability to sucrose was reversibly reduced by acidic pH. In addition, the pH effect on the permeability to sucrose fit well with by the Hill's equation (where, n=2.7 and pK=6.7).

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녹차색소의 특성과 염색성 (제1보) -녹차색소의 성분과 특성- (Characteristics and Dyeing Properties of Green Tea Colorants(Part I) -Components and characteristics of Green Tea Colorants-)

  • 신윤숙;최희
    • 한국의류학회지
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    • 제23권1호
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    • pp.140-146
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    • 1999
  • Colorants in green tea were extracted freeze-dried and analyzed to investigate the possibility of using as a natural dye. Fractionation of the colorants was carried out by column chromatograpy. Colorants in green tea were eluted into five fractions. All the fractions except fraction F2 showed absorption peakat 280nm. Fraction f2 showed absorption peak at 270nm and broad peak at 350nm, From the IR analysis it is speculated that fractions F2-F5 having similar stucture but different molecular weight are catechis. Silk fabrics dyes with fractions F1-F4 showed yellowish red color while sample dyed with fraction F5 showed red color. The colorants from green tea infusion was applied to silk wool nylon cotton and rayon fabrices. It showed relatively good affinity to protein and polyamide fibers bur low affinity to cellulose and regenerated cellulose fibers.

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Comparative Study of Korean (Viscum album var. coloratum) and European Mistletoes (Viscum album)

  • Lyu, Su-Yun;Park, Sun-Myo;Choung, Bo-Yun;Park, Won-Bong
    • Archives of Pharmacal Research
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    • 제23권6호
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    • pp.592-598
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    • 2000
  • A lectin (agglutinin, VCA) from Korean mistletoe (Viscum album L. coloratum) was isolated by affinity chromatograpy on a asialofetuin-Sepharose 4B. The molecular weights of A- and B-chains of VCA were different from those of VAAS. The VCA recognized the antibody of VAAs in the Western blot analysis and ELLA system. We also investigated the synergistic effects of the components in mistletoe by dividing the extract into different molecular weight fractions.

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