• Title/Summary/Keyword: fractionation.

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Reduction of eco-toxicity risk of heavy metals in the rotary drum composting of water hyacinth: Waste lime application and mechanisms

  • Singh, Jiwan;Kalamdhad, Ajay S.;Lee, Byeong-Kyu
    • Environmental Engineering Research
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    • v.20 no.3
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    • pp.212-222
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    • 2015
  • Experiments were conducted on the immobilization of eight heavy metals (HMs) (Zn, Cu, Mn, Fe, Ni, Pb, Cd, and Cr) during 20-day rotary drum composting of water hyacinth. The Tessier sequential extraction procedure was used to investigate the fractionation of HMs. The eco-toxicity risk of HMs was assessed by risk assessment code (RAC). In the results, the bioavailability factor (BAF) for different HMs presented in the following order: Mn > Zn = Fe > Cu > Cr > Cd = Pb > Ni. The total concentration of Pb was higher than that of Zn, Cu, Mn, Cd and Cr; however, its BAF was the lowest among these HMs. These results confirmed that the eco-toxicity of HMs depends on bioavailable fractions rather than on the total concentration. The greatest reduction in bioavailability and eco-toxicity risk of HMs occurred in lime 1% and 2% as compared to control and lime 3%. The eco-toxicity risk of Fe, Ni, Pb, Cd and Cr was reduced from low risk to zero risk by rotary drum composting. These studies demonstrated the high efficiency of the rotary drum for degrading compost materials and for reducing the bioavailability and eco-toxicity risk of HMs during the composting process.

Purification and Characterization of Aryl Acylamidase from Pseudomonas sp. (Pseudomonas sp. Aryl Acylamidase의 정제 및 성질)

  • 황인균;방원기
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.413-419
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    • 1998
  • Aryl acylamidase [EC 3.5.1.13] present in an acetaminophen-assimilating Pseudomonas sp. has been purified to a homogeneity using series of ammonium sulfate fractionation, DEAE-Sephacel anion exchange, Phenyl-Sepharose CL-4B hydrophobic, and Sephadex G-100 gel-permeation chromatography. The molecular weight, which was estimated by gel-permeation filtration and sodium dodecyl sulfate polyacylamide gel electrophoresis, was about 57 kDa and 56 kDa, respectively, indicating that this enzyme is a monomeric protein. The optimum pH was 10.5 and the optimum temperature was 40$^{\circ}C$. After incubation of the enzyme at 50$^{\circ}C$ for 30 min, residual activity of the enzyme was 34% compared to its original activity. The Km values for acetaminophen and 4'-nitroactanilide were 0.10 mM and 0.11 mM, respectively.

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Fractionation of Endoprotease from Viscera of the Argentina Shortfin Squid Illex argentinus (원양산 오징어(Illex argentinus) 내장으로부터 Endoprotease의 분획)

  • Kim, Hye-Suk;Kim, Jin-Soo;Heu, Min-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.41 no.3
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    • pp.176-181
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    • 2008
  • To evaluate the effective use of endoprotease from squid viscera as a food processing aid, various methods of fractionating endoprotease from viscera of the Argentina shortfin squid (Illex argentinus) were evaluated. The endoprotease-positive fractions of each fractionation were fraction II (30-40%, w/w) with cold acetone, fraction IV (50-60% saturation) with ammonium sulfate, fraction UF with anion exchange chromatography, and fraction II (15-24 kDa) with gel filtration. The specific activities (approximately 25 U/mg) of the fractions using ammonium sulfate and gel filtration were higher than the others. Total azocaseinolytic activity and recovery of the positive fraction using gel filtration were 806.95U and 37.82%, respectively, and were the highest among the positive fractions. Based on the results, gel filtration was the most efficient method for fractionating endoprotease from the viscera of Illex argentinus.

Purification and Characteristics of Pullulanase from Bacillus cereus subsp. mycoides (Bacillus cereus subsp. mycoides가 생산하는 Pullulanase의 정제와 특성)

  • Chung, Man-Jae;Woo, Jeong-Suk;Cho, Dae-Sun;Lee, Myong-Yur;Park, Nam-Kyu
    • Microbiology and Biotechnology Letters
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    • v.22 no.1
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    • pp.73-79
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    • 1994
  • The optimum cultural temperature and time for the pullulanase production by Bacillus cereus subsp. mycoides were 35$\circ $C and 48 hrs, respectively. The addition of egg albumin and casein to the basal medium increased the enzyme production. The enzyme was purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. specific activity of the purified enzyme was 82.37 U/mg protein and yield of theenzyume activity was 62.1%. The purified enzuyme showed a single band on ployacrylamide disc gel electrophoresis and its molecular weight was estimated to be 66.,000 by SDS-polyacrylamide disc gel electrophoresis. The isoelcular point for the purified enzyme was pH 5.0. The optimum temperature and pH were 50$\circ $C and pH 6.5, respectively. The purified enzyme was stable below 40$\circ $C and in the pH range of 6.5~10.0 The pullulanase activity was greatly inhited by Ag$^{+}$, Hg$^{2+}$ and EDTA, and its heat stability was increased by the addition of Ca$^{2+}$. The tydrolysis product with the enzyme on pullulan was maltotriose.

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In vitro Antioxidant Activity and Anticancer Effects of The Extracts from Eleutherococcus senticosus Max

  • Kim, Myong-Jo;Kim, Na-Young;Kang, Won-Hee;Choi, Won-Cheol;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.4
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    • pp.269-272
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    • 2002
  • Ethanol and butanol fractionation of E. senticosus showed strong anti-oxidant activity, and methanol and water extracts also had high anti-oxidant activity. The anti-oxidant activities in ethanol and butanol fractionation were higher than or similar to those of ${\alpha}-tocopherol$. The cytotoxic effect of root extract of E. senticosus was evaluated on seven different human cancer cell lines, The extracts of leaf and stem of E. senticosus also had strong antioxidant activity, but the antioxidant activity in root extract was higher than those in leaf and stem extracts. Methanol, hexane, and aqueous fraction layer had much higher inhibitory activities on lipid peroxidation in rat liver microsomes compared with ${\alpha}-tocopherol$. The effect of root extract of E. senticosus was evaluated on six human cancer cell lines. The values of 50% growth inhibition $(GI_{50})$ for the extracts were mostly below $30{\mu}g/ml$, and the extracts are considered as active inhibitory compounds on cancer cells.

Purification and Characterization of Acc I Endonuclease (Acc I endonuclease의 정제와 효소적 특성에 관한 연구)

  • 강선철;유욱준
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.13-19
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    • 1985
  • Acc I endonuclease has been isolated from 300g (wet weight) cells of Acinetobacter calcoaceticus. The cells were broken by using French press at 20, 000p.s.i. After ammonium sulfate fractionation, the enzyme was further purified by heparin agarose, DEAE-sephades, Affi.-gel Blue, phosphocellulose, and hydroxylapatite column chromatography. The purified Acc I endonudlease has a single polypeptide species and its subunit molecular weight was 45,000 ${\pm}$ 1,000 daltons as judged by 10% SDS-polyacrylamide gel electrophoresis. The isolated enzyme was essentially free of contaminating nucleases as judged by homochromatography by using a $^{32}P-labeled$ oligonucleotide. The enzyme showed maximum activity at pH values between 8.0 and 11.0 and in the presence of $MgCl_2$. Acc I endonuclease was maximally active in the absence of NaCl and was completely inhibited at 200 mM NaCl.

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Isolation of 3,4-Dihydroxybenzoic Acid, Which Exhibits Antimicrobial Activity, from Fruits of Gardenia jasminoides Ellis (치자 열매에서 항미생물 활성을 갖는 3,4-Dihydroxybenzoic Acid의 분리)

  • Yim, Cheol-Keun;Moon, Jae-Hak;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1386-1391
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    • 1999
  • The methanol extract of Gardenia jasminoides Ellis showed antimicrobial activity against bacteria and yeasts. The extract was successively purified with solvent fractionation, silica gel adsorption column chromatography, Sephadex LH-20 column chromatography, octadecylsilane column chromatography. The purified active substance was isolated by high performance liquid chromatography. The isolated compound was 3,4-dihydroxybenzoic acid which was determined by mass spectrometer, gas chromatograph-mass spectrometer, $^{1}H-nuclear$ magnetic resonance, $^{13}C-nuclear$ magnetic resonance and two-dimensional nuclear magnetic resonance. The content of 3,4-dihydroxybenzoic acid was $32.7\;{\mu}g/g$ in dried fruits of Gardenia jasminoides Ellis.

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Capillary Size-exclusion Chromatography as a Gel-free Strategy in Plasma Proteomics

  • Cho, Man-Ho;Wishnok, John S.;Tannenbaum, Steven R.
    • Molecular & Cellular Toxicology
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    • v.1 no.2
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    • pp.87-91
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    • 2005
  • Although 2D-PAGE has been widely used as the primary method for protein separation, difficulties in displaying proteins with an extreme values of isoelectric paint (pI), molecular size and hydrophobicity limit the technique. In addition, time consuming steps involving protein transfer and extraction from the gel-pieces can result in sample loss. Here, we describe a novel protein separation technique with capillary size-exclusion chromatography (CSEC) for rapid protein identification from human plasma. The method includes protein fractionation along with molecular size followed by in-solution tryptic digestion and peptide analysis through reversed phase liquid chromatography (RPLC) coupled to nanoflow electrospray-tandem mass spectrometry (ESI-MS/MS). Tryptic peptides are applied an a $100\;{\mu}m\;i.d.{\times}10mm$ length pre-column and then separated on a $75\;{\mu}m{\times}200mm$ analytical column at -100 nL/min flaw rate. Proteins were identified over the wide ranges of pI (3.7-12.3) when this technique was applied to the analysis of $1-2\;{\mu}L$ of human plasma. This gel-free system provides fast fractionation and may be considered a complementary technique to SDS-PAGE in proteomics.

An Acetylcholinesterase Inhibitor Isolated from Corydalis Tuber and Its Mode of Action (현호색의 Acetylcholinesterase 활성 저해 성분 및 그 작용기전)

  • Hwang, Se-Young;Chang, Young-Pyo;Byun, Soon-Jung;Jeon, Mee-Hee;Kim, Young-Choong
    • Korean Journal of Pharmacognosy
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    • v.27 no.2
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    • pp.91-95
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    • 1996
  • In the course of searching for acetylcholinesterase inhibitors from crude drugs, it was found that total MeOH extract of Corydalis Tuber showed significant inhibitory effect on acetylcholinesterase. To isolate acetylcholinesterase inhibitors from Corydalis Tuber, total MeOH extract of the the crude drug was subjected to activity guided fractionation. The MeOH extract was suspended in water and fractionated with methylene chloride and subjected to acid-base fractionation. Silica gel column chromatography of the basic fraction which showed significant inhibitory effect on acetylcholinesterase was carried out and 5 subfractions (1-5) were obtained. From subtraction 4, compound I was isolated. The structure of isolated compound I was identified by spectroscopic parameters of $^1H-NMR$, $^{13}C-NMR$, EI-MS and FAB-MS. The compound I was identified as berberine. It was found from the Lineweaver-Burk plot that berberine was a reversible and specific inhibitor of acetylcholinesterase having 90% inhibitory effect at the concentration of $2.5{\mu}M$.

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Oxalic Acid-based Remediation of Arsenic-contaminated Soil (옥살산 기반의 비소오염토양 정화 연구)

  • Lee, Myeong Eun;Jeon, Eun-Ki;Kim, Jong-Gook;Baek, Kitae
    • Journal of Soil and Groundwater Environment
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    • v.23 no.1
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    • pp.85-91
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    • 2018
  • Arsenic (As) usually is bound to amorphous iron oxides in the soils, and it can be removed via dissolution of iron oxides. Inorganic acid and chelating agent are widely used to extract As in the soil washing. However, the overall performance is highly dependent on the state of As fractionation. In this study, oxalic acid and inorganic acids (HCl, $H_2SO_4$, and $H_3PO_4$) were applied to enhance the dissolution of iron oxides for remediation of As-contaminated soils. Oxalic acid was most effective to extract As from soils and removal of As was two times greater than other inorganic acids. Additionally, 75% of As bound to amorphous iron oxides was removed by 0.2 M oxalic acid. Arsenic removal by oxalic acid was directly proportional to the sum of labile fractions of As instead of the total concentration of As. Therefore, the oxalic acid could extract most As bound to amorphous iron oxides.