• Title/Summary/Keyword: Ion gel

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Production and Purification of Single Chain Human Insulin Precursors with Various Fusion Peptides

  • Cho, Chung-Woo;Park, Sun-Ho;Nam, Doo-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.2
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    • pp.144-149
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    • 2001
  • For the production and purification of a single chain human insulin precursor, four types of fusion peptides $\beta$-galactosidase (LacZ), maltose binding protein (MBP), glutathione-S-transferase (GST), and (His)(sub)6-tagged sequence (HTS) were investigated. Recombinant E. coli harboring hybrid genes was cultivated at 37$\^{C}$ for 1h, and gene induction occurred when 0.2mM of isopropyl-D-thiogalactoside (IPTG) was added to the culture broth, except for E. coli BL21 (DE3) pLysS harboring a pET-BA cultivation with 1.0mM IPTG, followed by a longer than 4h batch fermentation respectively. DEAE-Sphacel and Sephadex G-200 gel filtration chromatography, amylose affinity chromatography, glutathione-sepharose 4B affinity chromatography, and a nickel chelating affinity chromatography system as a kind of immobilized metal ion affinity chromatography (IMAC) were all employed for the purification of a single chain human insulin precursor. The recovery yields of the HTS-fused, GST-fused, MBP-fused, and LacZ-fused single chain human insulin precursors resulted in 47%, 20%, 20%, and 18% as the total protein amounts respectively. These results show that a higher recovery yield of the finally purified recombinant peptides was achieved when affinity column chromatography was employed and when the fused peptide had a smaller molecular weight. In addition the pET expression system gave the highest productivity of a fused insulin precursor due to a two-step regulation of the gene expression, and the HTS-fused system provided the highest recovery of a fused insulin precursor based on a simple and specific separation using the IMAC technique.

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Purification and Biophysical Characterization of New Lectin from Baby Clam, Tapes japonica (바지락(Tapes japonica)으로부터 분리정제된 새로운 렉틴의 생물물리학적 특성)

  • Kim, Hee-Sook
    • Korean Journal of Food Science and Technology
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    • v.21 no.5
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    • pp.606-612
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    • 1989
  • A New lectin from baby clam, Tapes japonica, was isolated and purified through the following procedures; acetone powder, 0.15M NaCl extraction, ammonium sulfate fractionation, N-acetyl-D-galactosamine-agarose affinity column, and ion exchange Mono Q of FPLC. This lectin nonspecifically agglutinated human erythrocytes but didn't agglutinate mouse and rabbit erythrocytes. And the lectin neither stimulated human lymphocytes nor agglutinated Sarcoma 180 cells. On polyacrylamide gel electrophoresis, the lectin migrated as a major single band indicating homogeneous. A molecular weight was estimated to be about 131,000 daltons by Biogel P-300 and 125,000 daltons by SDS-PAGE without ${\beta}-mercaptoethanol$. This lectin is supposed to be a tetramer composed of heterogeneous subunits, about 30,000 and 33,000 daltons. Baby clam lectin was inhibited by EDTA and recovered agglutinating activity by $Ca^{++}\;and\;Mn^{++}$. This lectin is revealed as glycoprotein that contained about 4.2% neutral sugar.

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Purification and Characterization of an Intracellular NADH: Quinone Reductase from Trametes versicolor

  • Lee, Sang-Soo;Moon, Dong-Soo;Choi, Hyoung-T.;Song, Hong-Gyu
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.333-338
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    • 2007
  • Intracellular NADH:quinone reductase involved in degradation of aromatic compounds including lignin was purified and characterized from white rot fungus Trametes versicolor. The activity of quinone reductase was maximal after 3 days of incubation in fungal culture, and the enzyme was purified to homogeneity using ion-exchange, hydrophobic interaction, and gel filtration chromatographies. The purified enzyme has a molecular mass of 41kDa as determined by SDS-PAGE, and exhibits a broad temperature optimum between $20-40^{\circ}C$, with a pH optimum of 6.0. The enzyme preferred FAD as a cofactor and NADH rather than NADPH as an electron donor. Among quinone compounds tested as substrate, menadione showed the highest enzyme activity followed by 1,4-benzoquinone. The enzyme activity was inhibited by $CuSO_4,\;HgCl_2,\;MgSO_4,\;MnSO_4,\;AgNO_3$, dicumarol, KCN, $NaN_3$, and EDTA. Its $K_m\;and\;V_{max}$ with NADH as an electron donor were $23{\mu}M\;and\;101mM/mg$ per min, respectively, and showed a high substrate affinity. Purified quinone reductase could reduce 1,4-benzoquinone to hydroquinone, and induction of this enzyme was higher by 1,4-benzoquinone than those of other quinone compounds.

The Preparation and Characteristics of Covalently Cross-Linked SPEEK/Cs-TPA/Ceria Composite Membranes for Water Electrolysis (수전해용 공유가교 SPEEK/Cs-TPA/Ceria 복합막의 제조 및 특성 연구)

  • Song, Minah;Ha, Sungin;Park, Deayong;Ryu, Cheolhwi;Moon, Sangbong;Kang, Ansoo;Chung, Janghoon
    • Journal of Hydrogen and New Energy
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    • v.23 no.5
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    • pp.437-447
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    • 2012
  • Ceria ($CeO_2$) was used to scavenge free radicals which attack the membrane in the polymer electrolyte membrane water electrolysis (PEMWE) circumstance and to increase the duration of the membrane. In order to improve the electrochemical, mechanical and electrocatalytic characteristics, engineering plastic of the sulfonated polyether ether ketone (SPEEK) as polymer matrix was prepared in the sulfonation reaction of polyether ether ketone (PEEK) and the organic-inorganic blended composite membranes were prepared by sol-gel casting method with loading the highly dispersed ceria and cesium-substituted tungstophosphoric acid (Cs-TPA) with cross-linking agent contents of 0.01 mL. In conclusion, CL-SPEEK/Cs-TPA/ceria (1%) membrane showed the optimum results such as 0.130 S/cm of proton conductivity at $80^{\circ}C$, 2.324 meq./g-dry-membrane of ion exchange capacity and mechanical characteristics, and 65.03 MPa of tensile strength which were better than Nafion 117 membrane.

Synthesis and Characterization of Spherical SiO2@Y2O3 : Eu Core-Shell Composite Phosphors (구형 SiO2@Y2O3: Eu 코어-쉘 복합체 형광체 합성 및 특성)

  • Song, Woo-Seuk;Yang, Hee-Sun
    • Journal of the Korean Ceramic Society
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    • v.48 no.5
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    • pp.447-453
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    • 2011
  • The monodisperse spherical $SiO_2$ particles were overcoated with $Y_2O_3:Eu^{3+}$ phosphor layers via a Pechini sol-gel process and the resulting $SiO_2@Y_2O_3:Eu^{3+}$ core-shell phosphors were subsequently annealed at $800^{\circ}C$ at an ambient atmosphere. The crystallographic structure, morphology, and luminescent property of core-shell structured $SiO_2@Y_2O_3:Eu^{3+}$ phosphors were characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), and photoluminescence (PL). The spherical, nonagglomerated $SiO_2$ particles prepared by a Stober method exhibited a relatively narrow size distribution in the range of 260-300 nm. The thickness of phosphor shell layer in the core-shell particles can be facilely controlled by varying the coating number of $Y_2O_3:Eu^{3+}$ phosphors. The core-shell structured $SiO_2@Y_2O_3:Eu^{3+}$ phosphors showed a strong red emission, which was dominated by the $^5D_0-^7F_2$ transition (610 nm) of $Eu^{3+}$ ion under the ultraviolet excitation (263 nm). The PL emission properties of $SiO_2@Y_2O_3:Eu^{3+}$ phosphors were also compared with pure $Y_2O_3:Eu^{3+}$ nanophosphors.

Effect of Thin-Film Thickness on Electrical Performance of Indium-Zinc-Oxide Transistors Fabricated by Solution Process (용액 공정을 이용한 IZO 트랜지스터의 전기적 성능에 대한 박막 두께의 영향)

  • Kim, Han-Sang;Kyung, Dong-Gu;Kim, Sung-Jin
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.30 no.8
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    • pp.469-473
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    • 2017
  • We investigated the effect of different thin-film thicknesses (25, 30, and 40 nm) on the electrical performance of solution-processed indium-zinc-oxide (IZO) thin-film transistors (TFTs). The structural properties of the IZO thin films were investigated by atomic force microscopy (AFM). AFM images revealed that the IZO thin films with thicknesses of 25 and 40 nm exhibit an uneven distribution of grains, which deforms the thin film and degrades the performance of the IZO TFT. Further, the IZO thin film with a thickness of 30 nm exhibits a homogeneous and smooth surface with a low RMS roughness of 1.88 nm. The IZO TFTs with the 30-nm-thick IZO film exhibit excellent results, with a field-effect mobility of $3.0({\pm}0.2)cm^2/Vs$, high Ion/Ioff ratio of $1.1{\times}10^7$, threshold voltage of $0.4({\pm}0.1)V$, and subthreshold swing of $0.7({\pm}0.01)V/dec$. The optimization of oxide semiconductor thickness through analysis of the surface morphologies can thus contribute to the development of oxide TFT manufacturing technology.

Identification of Ligand for Salivary Lipocalin Secreted from the Uterine Endometrium during Early Pregnancy in Pigs

  • Seo, Hee-Won;Kim, Min-Goo;Ka, Hak-Hyun
    • Journal of Embryo Transfer
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    • v.24 no.4
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    • pp.259-263
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    • 2009
  • Salivary lipocalin (SAL1) is a member of the lipocalin protein family that has a property to associate with many lipophilic molecules and was identified as pheromone-binding protein in pigs. Our previous study has shown that SAL1 is expressed in the uterine endometrium in a cell type- and implantation stage-specific manner and secreted into the uterine lumen in pigs. However, function of SAL1 in the uterus during pregnancy in pigs is still not known. To understand physiological function of SAL1 in the uterine endometrium during pregnancy in pigs, it needs to elucidate the ligand(s) for SAL1. Thus, to identify the ligand for SAL1 in the porcine uterus, we collected uterine luminal fluid from pigs on day 12 of pregnancy by flushing with PBS. Proteins from the uterine luminal fluid were separated by ion exchange chromatography and gel filtration. Fractions containing SAL1 protein were pooled and concentrated. Immunoblot analysis confirmed successful purification of SAL1. Then, we extracted lipids from the purified SAL1 protein and analyzed the lipids by liquid chromatography-mass spectrometry, and predicted to be steroid hormones and prostaglandins as SAL1 ligands. Results in this study showed that SAL1 protein in the uterine secretions has a small lipophilic molecule as a natural ligand. Further characterization of ligand extracted from purified SAL1 will be useful for understanding physiological function of SAL1 during pregnancy and its application to increase the pregnancy rate in pigs.

Effect of Steam Curing on Compressive Strength of Slag Binder Concrete (증기양생이 고로슬래그 콘크리트의 압축강도에 미치는 영향)

  • Lim, Byung-Hoon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.8
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    • pp.338-343
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    • 2017
  • In this study, blast furnace slag powder was used in concrete to help reduce carbon dioxide emissions and to recycle industrial waste. Blast furnace slag powder is a byproduct of smelting pig iron and is obtained by rapidly cooling molten high-temperature blast furnace slag. The powder has been used as an admixture for cement and concrete because of its high reactivity. Using fine blast furnace slag powders in concrete can reduce hydration heat, suppress temperature increases, improve long-term strength, improve durability by increasing watertightness, and inhibit corrosion of reinforcing bars by limiting chloride ion penetration. However, it has not been used much due to its low compressive strength at an early age. Therefore, this study evaluates the effects of steam curing for increasing the initial strength development of concrete made using slag powder. The relationship between compressive strength, SEM observations, and XRD measurements was also investigated. The concrete made with 30% powder showed the best performance. The steam curing seems to affect the compressive strength by destroying the coating on the powder and by producing hydrates such as ettringite and Calcium-Silicate-Hydrate gel.

AN EXPERIMENTAL STUDY ON BOND STRENGTH OF COMPOSITE RESIN TO BLEACHED ENAMEL (표백된 법랑질에 대한 복합레진의 결합강도에 관한 연구)

  • Yu, Mi-Kyung;Lee, Kwang-Won;Song, Kwang-Yeob;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.19 no.1
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    • pp.114-123
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    • 1994
  • The purpose of this study was to examine the shear bond strength of resin-enamel bond formed at specific time intervals after the termination ov vital bleaching. A total of 72 human extracted maxillary premolars were divided into nine groups : untreated control (group 1) ; enamel treated with 35% hydrogen peroxide(group 2, 3, 4, 5) ; and enamel reated with 15% carbamide peroxide gel (group 6, 7, 8, 9). After the treatment with 35% hydrogen peroxide for 2 hours and 15% carbamide peroxide for 24 hours, adhesion of a resin to bleached enamel was formed at 1 hour (group 2, 6) and 24 hours(group 3, 7) ; 3days(group 4, 8) and 7 days(group 5, 9) post-termination of bleaching treatment. A $3{\times}3mm$ mold was filled with Scotchbond Multi-Purpose and Z100. After 24 hours later, the specimens were shear-tested at crosshead speed 1mm/min and analyzed statistically. Fractured specimens from group 1,2, 6 were gold-coated with Eiko ion coater and observed under Scanning electron microscope at 25KV. The following results results were obtained : 1. Bonds formed at 1 hour post-termination of 35 % hydrogen peroxide(P<0.01) and 15 % carbamide peroxide bleaching treatment groups(P<0.05) showed significantly lower shear bond strength than untreated group. 2. Bonds formed at 24 hours, 3 days and 7 days post-termination of 35% hydrogen peroxide and 15 % carbamide peroxide bleaching treatment groups showed no significant differences in shear bond strength with untreated group(p>0.05). 3. SEM examinations of the untreated fracture specimen indicated cohesive fracture within enamel and exposed enamel prisms, but the bleached fracture specimens indicated adhesive fracture.

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Study on Thermal Properties of CdS - Embedded Poly(2-Acetamidoacrylic acid) Hydrogel Composite (CdS 나노입자틀 삽입한 Poly(2-Acetamidoacrylic acid) 수화젤 복합체의 열적 특성에 관한 연구)

  • Park, Chun-Ho;Ha, Eun-Ju;Jung, Jong-Mo;Lee, Jang-Oo;Paik, Hyun-Jong
    • Polymer(Korea)
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    • v.33 no.1
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    • pp.1-6
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    • 2009
  • We report the template-based synthesis of well-dispersed CdS nanoparticles (NPs) in the interior of poly (2-acetamidoacrylic acid) (PAAA) hydrogel as a novel type of nanocomposite without particle aggregation via ion exchange in a aqueous system. As revealed by the TEM image analysis, the mean crystallite diameter of CdS NPs embedded in hydrogel composite was 4.5 nm, and the composite did not suffer any observable change after 6 months. Desorption/decomposition of CdS/PAAA hydrogel composite was studied by evolved gas analysis-gas chromatography-mass spectrometry (EGA-GC-MS), and thermogravimetric analysis (TGA) methods. From the TGA data, the thermal stability of the composite system increased by ca. 100 $^\circ$C and the content of CdS NPs in a dry composite gel was over 70 wt%. In addition, the chemical pathway was proposed for the entire decomposition process.