• Title/Summary/Keyword: ion exchange reaction

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Purification and Biological Characterization of Wild-type and Mutants of a Levan Fructotransferase from Microbacterium sp. AL-210 (Microbacterium sp. A-210이 생성하는 Levan fructotransferase의 정제 및 생물학적 특성에 관한 연구)

  • Hwang, Eun-Young;Jeong, Mi-Suk;Cha, Jae-Ho;Jang, Se-Bok
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1218-1225
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    • 2009
  • Difractose anhydrides (DFAs) is studied as a sweetener for diabetics because of its structural property. DFAs have four types: DFA I, III, IV (degradation of levan) and V (degradation of inulin). Especially, DFA IV has been shown to enhance the absorption of calcium in experiments using rats. Levan fructotransferase is an enzyme for producing di-d-fructose-2,6':6,2-dianhydride (DFA IV). To identify structural characterization, we purified wild-type and mutants (D63A, D195N and N85S) of levan fructotransferase (LFTase) from Microbacterium sp. AL-210. These proteins were purified to apparent homogeneity by Ni-NTA affinity column, Q-sepharose ion exchange and gel filtration chromatography and detected by SDS-PAGE. They were also analyzed by circular dichroism (CD) measurements, JNET secondary structure prediction, activity measurements at various temperatures, and pH analysis. The optimum pH for the enzyme-catalyzed reaction was pH 7.5 and optimum temperature was observed at $55^{\circ}C$. Along with wild-type LFTase, mutants were analyzed by CD measurement, fluorescence analysis and differential scanning calorimetry (DSC). N85S showed less $\alpha$-helix and more $\beta$ strand than others. Also, N85S showed almost the same curve as wild-type in their steady-state fluorescence spectra, whereas mutant D63A and D195N showed higher intensity than wild-type. The amino acid sequence of wild-type LFTase was compared to the sequences of exo-inulinase from Aspergillus awamori, a plant fructan 1-exohydrolase from Cichorium intybus, and Thermotogo maritime (Tm) invertase and showed a high identity with Exo-inulinase from Aspergillus awamori.

Evaluation of Denitrification Reactivity by the Supported Nanoscale Zero-Valent Iron Prepared in Ethanol-Water Solution (이중용매에서 제조된 나노영가철을 이용한 질산성질소의 환원반응성 평가)

  • Park, Heesu;Park, Yong-Min;Oh, Soo-Kyeong;Lee, Seong-Jae;Choi, Yong-Su;Lee, Sang-Hyup
    • Korean Chemical Engineering Research
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    • v.46 no.5
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    • pp.1008-1012
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    • 2008
  • Nanoscale zero-valent iron(nZVI) is famous for its high reactivity originated from its high surface area and it has received considerable attentions as one of the latest innovative technologies for treating contaminated groundwater. Due to its fine powdery form, nZVI has limited filed applications. The efforts to overcome this shortcoming by immobilizing nZVI on a supporting material have been made. This study investigated the differences of resin-supported nZVI's characteristics by changing the preparation methods and evaluated its reactivity. The borohydride reduction of an iron salt was proceeded in ethanol/water solvent containing a dispersant and the synthesis was conducted in the presence of ion-exchange resin. The resulting material was compared to that prepared in a conventional way of using de-ionized water by measuring the phyrical and chemical characteristics. BET surface area and Fe content of nZVI-attached resin was increased from $31.63m^2/g$ and 18.19 mg Fe/g to $38.10m^2/g$ and 22.44 mg Fe/g, respectively, by switching the solution medium from water to ethanol/water with a dispersant. The reactivity of each material was tested using nitrate solution without pH control. The pseudo first-order constant of $0.462h^{-1}$ suggested the reactivity of resin-supported nZVI prepared in ethanol/water was increased 61 % compared to that of the conventional type of supported nZVI. The specific reaction rate constant based on surface area was also increased. The results suggest that this new supported nZVI can be used successfully in on-site remediation for contaminated groundwater.