• Title/Summary/Keyword: anion exchange column

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Determination of 3-Butenyl Isothiocyanate in Different Parts and Cultivars of Chinese Cabbages

  • Kim, Youn-Kyung;Kim, Gun-Hee
    • Food Science and Biotechnology
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    • v.14 no.4
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    • pp.466-469
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    • 2005
  • Chinese cabbage (Brassica campestris L. Pekinensis) is Brassica vegetable that contains high amounts of glucosinolates. Glucosinolates and their breakdown products are thought to contribute to health promotion by preventing some cancers. Chinese cabbage is the most commonly consumed vegetable in Asian countries including Korea. In this study, qualitative and quantitative analyses of 3-butenyl glucosinolate (Gluconapin) from different cultivars and different parts of the cabbage were performed. Gluconapin of Chinese cabbage was extracted by hot ethanol ($80^{\circ}C$), isolated by an anion exchange column and identified by GC/MS and LC/MS. The levels of glucosinolates in Chinese cabbage varied according to the different parts, cultivars, and blanching time. In general, the concentrations of 3-butenyl isothiocyanate (ITC) were higher in the leaf than in the midribs parts. The cultivar 'Bulam no. 3' had a much greater content of 3-butenyl ITC than the cultivar 'Garak no. 1,' and the levels of butenyl ITC were highest after two weeks of storage. Blanching treatment decreased the concentration of 3-butenyl ITC. The ITC concentration varied extensively among different crops of the same species, and according to the different parts on the cabbage, the storage duration and the boiling time.

Biochemical Properties of a Chitin-Binding Class III Chitinase in Pumpkin Leaves

  • Lee, Kyun-Oh;Kim, Min-Gab;Jang, Ho-Hee;Lee, Ji-Yeun;Kim, Sun-Chang;Lee, Sang-Yeol
    • BMB Reports
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    • v.32 no.6
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    • pp.541-546
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    • 1999
  • When we compared the chitinase activity of various plant sources using colorimetric or active gel-staining assay methods, the specific activity of pumpkin leaves was the highest among the samples we analyzed. The highly active chitinase from pumpkin leaves (designated PL-ChtIII) was purified to homogeneity using affinity chitin gel and HPLC Mono-Q anion-exchange cloumn chromatographies. In contrast to other members of the class III chitinase family, PL-ChtIII showed a strong binding affinity to the regenerated chitin gel column. The apparent molecular weight of PL-ChtIII was estimated to be 29 kDa on SDS-PAGE gel, while its optimum pH and temperature were shown to be pH 6.0 and $60^{\circ}C$, respectively. Analyzing the reaction products of PL-ChtIII with swollen chitin as substrate, the dimer and tetramer of N-acetylglucosamine were produced as major products in the first hour of the enzymatic reaction along with a small amount of monomers and trimers. As the reaction time increased, dimeric N-acetylglucosamine became the predominant form of reaction product.

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Characterization of an Elastase Inhibitor Produced by Streptomyces lavendulae SMF11

  • Lee, Hyun-Sook;Jin, Wook;Kang, Sung-Gyun;Hwang, Yoon-Sook;Kho, Yung-Hee;Lee, Kye-Joon
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.81-85
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    • 2000
  • An elastase inhibitor, SMFEI02, was isolated from culture broth of Streptomyces lavendulae SMF11. The inhibitor was purified by ultrafiltration followed by XAD-7 column and Dowex-1 anion-exchange chromatographies, and preparative HPLC. The molecular formula was determined to be $C_{14}H_{16}N_2O_2$ (MW244) by HRFAB-MS analysis. The inhibitor was identified to be a diketopiperazine cyclo(S-Phe-S-Pro) by the optical rotation value and MNR spectral data, and showed inhibitory activities for trypsin, chymotrypsin, cathepsin B, and papain as well as elastase with the Ki values ranging from 1.78mM to $2.86{\;}\mu\textrm{m}$. The inhibition showed a competitive mode for elastase, chymotrypsin, and cathepsin B, whereas it showed a noncompetitive mode for trypsin and papain.

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Downstream Processing of Recombinant Hirudin Produced in Saccharomyces cerevisiae

  • Chung, Bong-Hyun;Kim, Won-Kyung;Rao, K.Jagannadha;Kim, Chul-Ho;Rhee, Sang-Ki
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.179-183
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    • 1999
  • A recombinant form of hirudin, a potent thrombin-specific inhibitor derived from the bloodsucking leech, was expressed as a secretory product in Saccharomyces cerevisiae under the control of GALl0 promoter and the mating factor $\alpha$pre-pro leader sequence. In an attempt to produce recombinant hirudin (r-Hir) of therapeutic purity in large quantities, the fed-batch fermentation was carried out by using this recombinant yeast, and subsequently downstream processing was developed with the preparative-scale column chromatography systems. About 234 mg/l of biologically active r-Hir was produced as a secretory product by the fed-batch fermentation strategy developed for an efficient downstream processing. Using a two-step chromatography process (an anion exchange chromatography followed by the reverse phase HPLC), the r-Hir was purified to>98% with an overall recovery yield of 84%. According to the N-terminal amino acid sequencing, the purified r-Hir was found to have the predicted N-terminal amino acid sequence. The biological activity of the purified r-Hir to inhibit thrombin was also identical to that of the commercial hirudin.

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Antifungal and Anticancer Activities of a Protein from the Mushroom Cordyceps militaris

  • Park, Byung-Tae;Na, Kwang-Heum;Jung, Eui-Cha;Park, Jae-Wan;Kim, Ha-Hyung
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.1
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    • pp.49-54
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    • 2009
  • The mushroom Cordyceps militaris has been used for a long time in eastern Asia as a nutraceutical and in traditional Chinese medicine as a treatment for cancer patients. In the present study, a cytotoxic antifungal protease was purified from the dried fruiting bodies of C. militaris using anion-exchange chromatography on a DEAE-Sepharose column. Electrophoretic analyses indicated that this protein, designated C. militaris protein(CMP), has a molecular mass of 12 kDa and a pI of 5.1. The optimum conditions for protease activity were a temperature of $37^{\circ}C$ and pH of $7.0{\sim}9.0$. The enzyme activity was specifically inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride. Amino acid composition of intact CMP and amino acid sequences of three major peptides from a tryptic digest of CMP were determined. CMP exerted strong antifungal effect against the growth of the fungus Fusarium oxysporum, and exhibited cytotoxicity against human breast and bladder cancer cells. These results indicate that C. militaris represents a source of a novel protein that might be applied in diverse biological and medicinal applications.

Separation of Pu and Nd from Uranium Matrix by Equilibrated Cation Exchanger for Burnup Measurement of Irradiated Nuclear Fuel (조사후핵연료의 연소도 측정을 위한 동적이온교환체에 의한 우라늄 매질로부터 Pu 및 Nd의 분리)

  • Joe, Kih-Soo;Kim, Jung-Suk;Jeon, Young-Shin;Han, Sun-Ho;Eom, Tae-Yoon
    • Nuclear Engineering and Technology
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    • v.25 no.2
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    • pp.259-264
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    • 1993
  • Ion chromatographic method has been applied for burnup measurement of irradiated nuclear fuel by dynamic system using 1-octanesulfonate as a cation exchanger and $\alpha$-hydroxyisobutyric acid as an eluant. A number of elution techniques were evaluated for the optimum separation of plutonium, uranium and neodymium. These elements were individually separated and collected by gradient elution between 0.05 M and 0.40 M of $\alpha$-hydroxyisobutyric acid in a single column, and finally determined by isotope dilution mass spectrometry. The burnup data from this method were compared with those from conventional anion exchange method. The results showed a good agreement within 3.5 % of difference between two methods.

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Enzymatic N-glycan analysis of 31 kDa molecule in plerocercoid of Spirometra mansoni (sparganum) and its antigenicity after chemical oxidation

  • Chung, Young-Bae;Kong, Yoon;Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.42 no.2
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    • pp.57-60
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    • 2004
  • A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.

Flatfish Vitellogenin Detection Using Optical Waveguide Lightmode Spectroscopy-based Immunosensor

  • Kim, Nam-Soo;Ryu, Hyung-Seok;Kim, Woo-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1445-1451
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    • 2007
  • A sensitive optical waveguide lightmode spectroscopy-based immunosensor was developed to detect vitellogenin in seawater flatfish (Paralichthys olivaceus). For this purpose, anion-exchange column chromatography with DE-52 resin was used to purify flatfish vitellogenin from flatfish serum containing vitellogenin that had been induced using an intraperitoneal $17{\beta}$-estradiol injection. The anti-flatfish vitellogenin antibody used as the biological component of the above immunosensor was prepared using the purified flatfish vitellogenin. The change in the incoupling angle according to the complexation between the flatfish vitellogenin and its antibody, immobilized over an optical grating coupler sensor chip, was measured to calculate the sensor response. The immunosensor was quite specific to flatfish vitellogenin binding, based on no sensor response in the case of bovine serum albumin immobilization. When plotted using double-logarithmic scales, the sensor responses increased linearly in flatfish vitellogenin concentrations of 0.00675-67.5 nM, with a detection limit of 0.0675 nM. The reusability during seven repetitive measurements was reasonably fair for the preliminary screening of flatfish vitellogenin.

Enhanced removal of phosphate on modified ion exchanger with competing ion (음이온 교환수지를 이용한 인제거 향상)

  • Nam, Ju-Hee;Lee, Sang-Hyup;Choi, Jae-Woo;Hong, Seok-Won;An, Byungryul
    • Journal of Korean Society of Water and Wastewater
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    • v.27 no.1
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    • pp.121-128
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    • 2013
  • The concern for dissolved phosphate in water/wastewater has been increasing because of the risk for eutrophication. A variety of conventional and advanced technologies were applied to meet the enforced new regulation of phosphate around the world. However, there still remained a lot of challenge because most introduced/developed method, for example, biological and physic-chemical treatment is not easy to satisfy the new regulation of phosphate in water. In order to meet the new regulation, the application of ion exchanger has been tried which showed that the removal efficiency for phosphate was strongly determined by in the presence of the competing ion, especially sulfate. As results, a new class of ion exchanger governed by ligand exchange was developed and investigated to increase the selectivity for phosphate. The current study using organic/inorganic anion exchanger developed with Lewis acid-base interaction confirms the selectivity for phosphate over sulfate. According to isotherm test and column test, the value of the maximum phosphate uptake (Q) showed 64 mg/g as $po{_4}^{3-}$ and the breakthrough for phosphate occurs after 1000 min and completely finishes at 2500 min, respectively.

Purification of Anti-allergic Compound from Mori Cortex Radicis Extract (상백피 추출물 중의 항알러지 활성 성분의 분리)

  • Lee, Eun-Ju;Chae, Ok-Hee;Lee, Moo-Sam;Lee, Hern-Ku;Huh, Hoon
    • YAKHAK HOEJI
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    • v.42 no.4
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    • pp.395-402
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    • 1998
  • Mori Cortex Radicis, the root bark of mulberry tree, has been used in the treatment of bronchial asthma and other lung diseases in traditional medicine. There was a recent repor t that the water soluble part with molecular weight of above 10,000 has anti-allergic activity. Therefore, we intended to isolate and purify the anti-allergic compound from hot water extract of the Mori Cortex Radicis. Crude extract of Mori Cortex Radicis was prepared by hot-water extraction, and anti-allergic compound was further purified by alcohol precipitation, successive ultrafiltration, anion exchange chromatography and gel filtration chromatography. This compound had homogeneity which was shown by the sharp single peak in HPLC chromatogram (TSK-GEL G400OPW column, RI detector). The molecular weight of the compound was estimated as 23Kda on the basis of calibration curve plotted against protein standards. This compound was identified as complex of sugar, protein and lignin (19.2: 5.9: 72.7), and proteolysis could not decrease the anti-allergic activity but mild delignification decreased the activity remarkably. Therefore, we concluded that the anti-allergic compound of Mori Cortex Radicis was a lignin-carbohydrate complex.

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