• Title/Summary/Keyword: H-Q curve

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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.

Sorption Efficiency of the Bamboo Charcoal to Remove the Cesium in the Contaminated Water System (오염수계 내 세슘 제거를 위한 대나무 활성탄의 흡착효율 규명)

  • Ahn, Joungpil;Lee, Minhee
    • Economic and Environmental Geology
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    • v.51 no.2
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    • pp.87-97
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    • 2018
  • The cesium (Cs) removal from the contaminated water system has been considered to be difficult because the cesium likes to exist as soluble phases such as ion and complexes than the solid in water system. Many researches have focused on developing the breakthrough adsorbent to increase the cesium removal efficiency in water. In this study, the laboratory scale experiments were performed to investigate the feasibility of the adsorption process using the bamboo charcoal for the Cs contaminated water system. The Cs removal efficiency of the bamboo charcoal were measured and the optimal adsorption conditions were determined by the adsorption batch experiments. Total 5 types of commercialized bamboo charcoals in Korea were used to identify their surface properties from SEM-EDS and XRD analyses and 3 types of bamboo charcoals having large specific surface areas were used for the adsorption batch experiment. The batch experiments to calculate the Cs removal efficiency were performed at conditions of various Cs concentration (0.01 - 10 mg/L), pH (3 - 11), temperature ($5-30^{\circ}C$), and adsorption time (10 - 120 min.). Experimental results were fitted to the Langmuir adsorption isotherm curve and their adsorption constants were determined to understand the adsorption properties of bamboo charcoal for Cs contaminated water system. From results of SEM-EDS analyses, the surfaces of bamboo charcoal particles were composed of typical fiber structures having various pores and dense lamella structures in supporting major adsorption spaces for Cs. From results of adsorption batch experiments, the Cs-133 removal efficiency of C type bamboo charcoal was the highest among those of 3 bamboo charcoal types and it was higher than 75 % (maximum of 82 %) even when the initial Cs concentration in water was lower than 1.0 mg/L, suggesting that the adsorption process using the bamboo charcoal has a great potential to remove Cs from the genuine Cs contaminated water, of which Cs concentration is low (< 1.0 mg/L) in general. The high Cs removal efficiency of bamboo charcoal was maintained in a relatively wide range of temperatures and pHs, supporting that the usage of the bamboo charcoal is feasible for various types of water. Experimental results were similar to the Langmuir adsorption model and the maximum amount of Cs adsorption (qm:mg/g) was 63.4 mg/g, which was higher than those of commercialized adsorbents used in previous studies. The surface coverage (${\theta}$) of bamboo charcoal was also maintained in low when the Cs concentration in water was < 1.0 mg/L, investigating that the Cs contaminated water can be remediated up with a small amount of bamboo charcoal.