• Title/Summary/Keyword: Anion Exchange

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Purification and Characterization of Vitellin from the Firefly, Pyrocoelia rufa

  • Kim, Seong-Ryul;Jin, Byung-Rae;Yang, Won-Jin;Kim, Jong-Gill;Kim, Keun-Young;Lee, Sang-Mong;Moon, Byung-Ju;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.2
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    • pp.167-172
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    • 2001
  • The vitellin of firefly, Pyrocoelia rufa, is composed of three polypeptides, designated Vn1 (175 kDa), Vn2 (160 kDa) and Vn3 (45 kDa) in SDS-polyacrylamide gel electrophoresis. Three subunits of vitellin were presented in the female adult hemolymph, ovary and egg extracts, but not observed in the male. This vitellin was purified from the eggs of P. rufa by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. In nature, vitellin of P. rufa has molecular weight of 400 kDa. Western blot analysis using polyclonal antiserum against purified vitellin showed that the antiserum was reacted with the three polypeptides, Vnl, Vn2 and Vn3 from the female adult hemolymph, ovary and egg extracts. Amino acid residues at N-terminus of three subunits were sequenced. The N-terminal sequences of large subunits, Vnl and Vn2, were similar to each other, But, the N-terminal sequences of small subunits Vn3, did not have any signnificant homology with large subunits.

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Different Level of Plasma Free Hemoglobin between Qi-deficiency and Fire Heat among Korean Stoke Subjects (한국인 중풍환자의 기허군 화열군의 plasma free hemoglobin의 비교)

  • Lim, Ji-Hye;Ko, Mi-Mi;Lee, Jung-Sup;Lee, Myeong-Soo;Cha, Min-Ho
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.4
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    • pp.697-701
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    • 2011
  • The purpose of this study was to fine proteins, which have significantly different level in plasma between Qi-deficiency and Fire-heat group of Korean Oriental Stroke pattern identification (PI) among Korean stroke patients. Eighteen stroke patients with Qi-deficiency and forty nine patients with Fire-heat, which had critical syndrome of each PI, were participated in this study. Plasma protein pattern were analyzed by SELDI-TOF MS using Q10 strong anion exchange chip and Mass spectral data (m/z) statistically determined. The expression level of proteins, which were different between Qi-deficiency and Fire-heat in the results by SELDI-TOF MS, were confirmed by western blot. As a result of analyzing plasma protein by SELDI-TOF MS, six protein peaks were significantly higher in Fire-heat group than Qi-deficiency group. Two peaks among of them, M15003 and M15745, were respectively identified as hemoglobin alpha and beta in previous study. Expression level of plasma free hemoglobin of Fire-heat group was also confirmed higher in Fire-heat group than in Qi-deficiency group. These findings suggest that plasma free hemoglobin is a candidate for discriminating Qi-deficiency and Fire-heat group according to pattern identification (PI) of stroke.

Screening and Partial Purification of Haloperoxidase from Marine Actinomycetes (해양방선균으로부터 Haloperoxidase의 검색과 특성)

  • Cho, Ki-Woong
    • Korean Journal of Microbiology
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    • v.44 no.2
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    • pp.116-121
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    • 2008
  • In my search of microbial source of novel enzymes, a marine actinomycetes, A1460, producing haloperoxidase was isolated from macroalgae from south sea, Korea and studied for physiological and biochemical properties. The haloperoxidation reaction was followed by the bromination of phenol red in the presence of hydrogen peroxide and potassium bromide. The haloperoxidase was partially purified from the cell extract with $35\sim75%$ ammonium sulfate precipitation, High-Q anion exchange chromatography, gel filtration chromatography, hydroxyapetite chromatography and hydrophobic interaction chromatography to a yield of 42% and purification fold of 70. This enzyme showed relatively high heat stability without losing 50% of activity after 1 hr incubation at $60^{\circ}C$. The highest activity was found at $45^{\circ}C$, and the optimal pH was about pH 7, but higher stability was observed at pH 8. Azide and cyanide ion showed strong inhibition at less than 1 $\mu M$ level suggesting that the enzyme was Fe ion dependent haloperoxidase.

Determination of carbon-14 and tritium in a PWR spent nuclear fuel (PWR 사용후핵연료 중 탄소-14 및 트리튬 정량)

  • Kim, Jung Suk;Park, Soon Dal;Lee, Chang Hun;Song, Byong Chul;Jee, Kwang Yong
    • Analytical Science and Technology
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    • v.18 no.4
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    • pp.298-308
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    • 2005
  • The methods for determining C-14 and tritium contents in the spent nuclear fuel sample were developed. The carbon-14($^{14}CO_2$) released during the dissolution of the spent fuel sample and $CaCO_3$ ($CO_2$ carrier) with 8 M $HNO_3$ at $90^{\circ}C$ was collected in trap containing 1.5 M NaOH. The volatile radioactive iodine evolved when the spent fuel was dissolved, was trapped on to Ag-silicagel (Ag-impregnated silicagel) adsorbent in column which is connected to two NaOH traps. The solutions which contain tritium as HTO after fuel dissolution were decontaminated by deionization with a mixture of cation and anion exchange resins and inorganic ionexchangers. The amount of C-14 in the trap solutions and the HTO concentration in the resulting deionization water were then determined by liquid scintillation counting.

Purification and Characterization of Recombinant Human Follicle Stimulating Hormone Produced by Chinese Hamster Ovary Cells

  • NA KYU HEUM;KIM SEUNG CHUL;SEO KWANG SEOK;LEE SUNG HEE;KIM WON BAE;LEE KANG CHOON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.395-402
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    • 2005
  • Biologically active recombinant human follicle stimulating hormone (rhFSH) was produced in Chinese hamster ovary cells and purified by a series of chromatographic steps. The chromatographic steps included anion-exchange chromatography (DEAE Sepharose F/F, Q Sepharose F/F), hydrophobic interaction chromatography (Source 15 PHE), and hydroxyapatite chromatography (Macro-Prep ceramic hydroxyapatite type I). A distinctive step of the purification process developed was the use of ZnCl$_2$ for the removal of non-glycosylated or lowly-glycosylated FSH and impurities through co-precipitation with Zn$^{2+}$. Purified rhFSH was identified and characterized by several physicochemical and biological methods such as gel electrophoresis, high-performance liquid chromatography, amino acid analysis, carbohydrate analysis, and biological activity. The overall yield of the purification was ~$30\%$. The rhFSH preparation obtained showed high purity (>$99\%$) and high in vivo potency (>16,000 IU/mg). Carbohydrate analysis suggested that the purified rhFSH contained approximately $40\%$ (w/w) carbohydrate with di­or tri-antennary structure on average, which is somewhat more heavily sialylated than commercially available rhFSH. In conclusion, the results of these analyses established an identity of the purified rhFSH with natural FSH from human pituitary glands, and furthermore, the purified rhFSH preparation showed higher in vivo potency and was slightly more heavily sialylated than commercially available rhFSH.

Purification and Characterization of Chitinase from Paenibacillus illinoisensis KJA-424

  • JUNG WOO JIN;KUK JU HEE;KIM KIL YONG;KIM TAE HWAN;PARK RO DONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.274-280
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    • 2005
  • A chitinase was purified from the culture supernatant of Paenibacillus illinoisensis KJA-424 by protein precipitation, DEAE-Sephadex anion-exchange chromatography, and Sephadex G-150 gel filtration. The molecular weight of the purified chitinase was 54 kDa on SDS-PAGE and activity staining. Optimal pH and temperature were pH 5.0 and 60$^{circ}$C, the presence of 10 ruM Ag$^{+}$ and Hg$^{2+}$ inhibited the activity by $92.1/%$ and $97.7/%$, and the K$_{m}$ and V$_{max}$ values were 1.12 mg chitin mrl and 1.48$\mu$mol GlcNAc min$^{-1}$, respectively. The enzyme hydrolyzed tetramer to dimer, pentamer to dimer and trimer, and hexamer to dimer, trimer and tetramer, indicating an endo-splitting mechanism. The chitinase had no hydrolytic activity toward dimer and trimer. The chitinase inhibited the mycelial growth of Rhizoctonia solani, suggesting an antifungal property.

Linkage Structure Analysis of Barley and Oat $\beta$-Glucans by High Performance Anion Exchange Chromatography

  • Ryu, Je-Hoon;Yoo, Dong-Hyung;Lee, Byung-Hoo;Lee, Su-Yong;Joo, Mi-Hyun;Yoo, Sang-Ho
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.271-274
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    • 2009
  • Cereal $\beta$-glucans, linked essentially by mixed $\beta$-(1,4/1,3) glycosidic bonds, were extracted, purified, and structurally identified. Previously chemical structure of barley $\beta$-glucans was characterized from 3 varieties of 'Gang', 'Ohl', and 'Gwangan', and the (1,4)/(1,3) linkage ratio of the $\beta$-glucans was identical. In this study, $\beta$-glucans from 1 barley ('Chal') and 3 oat ('Ohl', 'Samhan', and 'Donghan') varieties were structurally scrutinized, and the linkage pattern of total 7 cereal $\beta$-glucans was compared. The amount of 2 major 3-O-$\beta$-cellobiosyl-D-glucose (DP3) and 3-O-$\beta$-cellotriosyl-D-glucose (DP4) from barley and oat accounted for only 66.6-73.3 and 68.12-81.89% of water-extractable $\beta$-glucan fractions, and the (1,4)/(1,3) linkage ratios of both barley and oat $\beta$-glucans were within very narrow range of 2.27-2.31 and 2.38-2.39, respectively, among the cultivars tested. Structural difference in the cereal $\beta$-glucans was evident when DP3:DP4 ratio in the $\beta$-glucan structure was compared. As a result, this ratio was significantly greater for barley $\beta$-glucan (2.26-2.74) than for oat (1.54-1.66). Chal-B had the greatest DP3 to DP4 ratio among the samples, which in turn reflected the least amount of (1,4)-linkages.

DETERMINATION OF THE TRANSURANIC ELEMENTS INVENTORY IN HIGH BURNUP PWR SPENT FUEL SAMPLES BY ALPHA SPECTROMETRY

  • Joe, Kih-Soo;Song, Byung-Chul;Kim, Young-Bok;Han, Sun-Ho;Jeon, Young-Shin;Jung, Euo-Chang;Jee, Kwang-Yong
    • Nuclear Engineering and Technology
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    • v.39 no.5
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    • pp.673-682
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    • 2007
  • The contents of transuranic elements in high-burnup spent fuel samples were determined. The activity amounts of $^{238}Pu,\;^{239}Pu,\;^{240}Pu,\;^{241}Am,\;^{244}Cm\;and\;^{242}Cm$ were measured by alpha spectrometry using $^{242}Pu\;and\;^{243}Am$ as tracers, respectively. A spike addition method for $^{237}Np$ was established by an alpha and gamma spectrometry using $^{239}Np$ as a spike after the optimum conditions for the measurements of $^{237}Np\;and\;^{239}Np$, respectively, were obtained. A separation system using anion exchange chromatography and diethylhexylphosphoric acid extraction chromatography was applied for the separation of these elements. This method was applied to high-burnup spent nuclear fuel samples $(40{\sim}60GWD/MTU)$. The contents of the transuranic elements were compared with those by ORIGEN-2 code. Measurements and the calculations of the contents of the plutonium isotopes $^{238}Pu,\;^{239}Pu\;and\;^{240}Pu$ agreed to within 10% on average. The contents of $^{237}Np$ agreed to within approximately 5% except for one instance of a calculation, while those of $^{241}Am,\;^{244}Cm\;and\;^{242}Cm$ showed higher values by approximately 19%, 35% and 14% on average, respectively, compared to the calculations according to the burnup.

Burnup Measurement of Spent $U_3$Si/Al Fuel by Chemical Method Using Neodymium Isotope Monitors

  • Kim, Jung-Suk;Jeon, Young-Shin;Park, Kwang-Soon;Song, Byung-Chul;Han, Sun-Ho;Kim, Won-Ho
    • Nuclear Engineering and Technology
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    • v.33 no.4
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    • pp.375-385
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    • 2001
  • The total burnup in the spent U$_3$Si/Al fuel samples from Hanaro reactor was determined by destructive methods using $^{148}$ Nd, the sum of $^{143}$ Nd and $^{144}$ Nd, the sum of $^{145}$ Nd and $^{146}$ Nd, and the sum of total Nd isotopes($^{143}$ Nd, $^{144}$ Nd, $^{145}$ Nd, $^{146}$ Nd, $^{148}$ Nd and $^{150}$ Nd) monitors. The fractional($^{235}$ U) turnup in the spent fuel samples was also determined by U and Pu mass spectrometric method. The samples were dissolved in a mixture of 4 M HCI and 10 M HNO$_3$ without any catalyst. The separation of U, Pu and Nd from the spiked and unspiked sample solutions was achieved by two sequential anion exchange separation methods. The isotope compositions of these elements, after their separation from the fuel samples were measured by mass spectrometry. The contents of the elements in the spent fuel samples were determined by isotope dilution mass spectrometric method(IDMS) using $^{233}$ U, $^{242}$ Pu and $^{150}$ Nd as spikes. The effective fission yield was calculated from the weighted fission yields averaged over the irradiation period. The difference between total turnup values determined by various Nd monitors were in the range of 1.8%.

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Biochemical Characteristics of Lectins Isolated from Lentinula edodes (표고버섯으로부터 분리한 렉틴의 생화학적 특성)

  • Kim, Young-Sin;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • v.29 no.4
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    • pp.79-88
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    • 2001
  • Lectin was isolated from shiitake mushroom (Lentinula edodes) with 0.15 M NaCl solution, and purified by the following procedures : precipitation by ammonium sulfate, anion exchange column chromatography on DEAE Sephadex A-50 and hydroxyapatite column chromatography. The fresh pileus part of the mushroom contained more than two times of lectin compared to the stipe part, and lectins and its activity were reduced by heating. The extraction yield of crude lectin was 46.03%, 28% yield after purification on on DEAE Sephadex A-50 column chromatography. Some amino acids, aspartic acid, serine, alanine and histidine, were increased by purification process. Relatively low molecular weight parts of lectin had the agglutinating activity for rabbit blood, and its molecular weight was about 23 kDa The molecular weights of purified lectins, LA-a and LB-b, by the hydroxyapatite column chromatography were 24 kDa and 23 kDa, respectively.

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