• Title/Summary/Keyword: Gel Layer

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Proteomic Analysis of Global Changes in Protein Expression During Exposure of Gamma Radiation in Bacillus sp. HKG 112 Isolated from Saline Soil

  • Gupta, Anil Kumar;Pathak, Rajiv;Singh, Bharat;Gautam, Hemlata;Kumar, Ram;Kumar, Raj;Arora, Rajesh;Gautam, Hemant K.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.574-581
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    • 2011
  • A Gram-positive bacterium was isolated from the saline soils of Jangpura (U.P.), India, and showed high-level of radiation-resistant property and survived upto 12.5 kGy dose of gamma radiation. The 16S rDNA sequence of this strain was examined, identified as Bacillus sp. strain HKG 112, and was submitted to the NCBI GenBank (Accession No. GQ925432). The mechanism of radiation resistance and gene level expression were examined by proteomic analysis of whole-cell extract. Two proteins, 38 kDa and 86.5 kDa excised from SDS-PAGE, which showed more significant changes after radiation exposure, were identified by MALDI-TOF as being flagellin and S-layer protein, respectively. Twenty selected 2-DE protein spots from the crude extracts of Bacillus sp. HKG 112, excised from 2- DE, were identified by liquid chromatography mass spectrometry (LC-MS) out of which 16 spots showed significant changes after radiation exposure and might be responsible for the radiation resistance property. Our results suggest that the different responses of some genes under radiation for the expression of radiation-dependent proteins could contribute to a physiological advantage and would be a significant initial step towards a fullsystem understanding of the radiation stress protection mechanisms of bacteria in different environments.

Maximizing TPBs through Ni-self-exsolution on GDC based composite anode in solid oxide fuel cells

  • Tan, Je-Wan;Lee, Dae-Hui;Kim, Bo-Gyeong;Kim, Ju-Seon;Mun, Ju-Ho
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.402.1-402.1
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    • 2016
  • The performance of solid oxide fuel cells (SOFCs) is directly related to the electrocatalytic activity of composite electrodes in which triple phase boundaries (TPBs) of metallic catalyst, oxygen ion conducting support, and gas should be three-dimensionally maximized. The distribution morphology of catalytic nanoparticle dispersed on external surfaces is of key importance for maximized TPBs. Herein in situ grown nickel nanoparticle onto the surface of fluorite oxide is demonstrated employing gadolium-nickel co-doped ceria ($Gd0.2-xNixCe0.8O2-{\delta}$, GNDC) by reductive annealing. GNDC powders were synthesized via a Pechini-type sol-gel process while maximum doping ratio of Ni into the cerium oxide was defined by X-ray diffraction. Subsequently, NiO-GNDC composite were screen printed on the both sides of yttrium-stabilized zirconia (YSZ) pellet to fabricate the symmetrical half cells. Electrochemical impedance spectroscopy (EIS) showed that the polarization resistance was decreased when it was compared to conventional Ni-GDC anode and this effect became greater at lower temperature. Ex situ microstructural analysis using scanning electron microscopy after the reductive annealing exhibited the exsolution of Ni nanoparticles on the fluorite phases. The influence of Ni contents in GNDC on polarization characteristics of anodes were examined by EIS under H2/H2O atmosphere. Finally, the addition of optimized GNDC into the anode functional layer (AFL) dramatically enhanced cell performance of anode-supported coin cells.

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Purification and Characterization of Xylanase from Bacillus sp. A-6 (Bacillus sp. A-6의 Xylanase의 정제와 특성)

  • Choi, Suk-Ho
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.147-152
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    • 2009
  • A xylanase was purified from the culture supernatant of Bacillus sp. A-6 by using ultrafiltration and ion exchange chromatography on the column of SP-Sepharose using 5 mM acetate buffer, pH 5.0. The xylanase was eluted from the column at the concentration less than 0.05 M NaCl. The eluted xylanase was shown to be a single protein band in SDS-PAGE. Zymogram analysis indicated that the protein band in SDS-PAGE had the enzyme activity to hydrolyze oat spelt xylan. The molecular weights of the xylanase were 15,000 based on SDS-PAGE and 14,100 based on gel filtration chromatography. Thin layer chromatography showed that the xylanase hydrolyzed oat spelt xylan into xylobiose and high-molecular-weight xylooligosaccharides. The relative activities of the heated xylanase decreased to 80% at $40^{\circ}C$ after 7 hr and less than 40% at $60^{\circ}C$ after 1 hr.

Biochemical Study on Soft-rotted Sweetpotatoes (연부(軟腐)고구마의 생화학적(生化學的) 연구(硏究))

  • Chun, Jae-Kun
    • Applied Biological Chemistry
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    • v.10
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    • pp.47-61
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    • 1968
  • Sweetpotato infected by Soft-rot, Rhizopus nigricans, has been investigated for its biochemical changes. The results of the present experiment are summarized as follows: 1. From the Soft-rotted tuber, nineteen Ehrlich's positive substances and three fluorescent compounds which had not been found in a healthy tuber were detected with thin layer chromatography. 2, Ipomeamarone and umbelliferone, each of which are predominating substances among the Ehrlich's positive and fluorescent compounds, were isolated by silica gel column chromatography and studied for their identities. 3. Time course biosynthesis of the metabolites was studied and discussed for their possible biosynthetic pathways. 4. Among the metabolites, ipomeamarone and ipomeamarone-like substances have enzyme: inhibitive action against ${\alpha}$-amylase and probably other enzymes; the inhibition type was determined as the uncompetitive one. 5. Ipomeamarone obtained from soft-rotted sweetpotato was proved to have an uncoupling action as 2, 4-dinitrophenol.

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Isolation and Characterization of an Acyclic Isoprenoid from Semecarpus anacardium Linn. and its Antibacterial Potential in vitro - Antimicrobial Activity of Semecarpus anacardium Linn. Seeds -

  • Purushothaman, Ayyakkannu;Meenatchi, Packirisamy;Saravanan, Nallappan;Karuppaiah, Muthu;Sundaram, Ramalingam
    • Journal of Pharmacopuncture
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    • v.20 no.2
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    • pp.119-126
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    • 2017
  • Objectives: Semecarpus anacardium Linn. is a plant well-known for its antimicrobial, antidiabetic and anti-arthritic properties in the Ayurvedic and Siddha system of medicine. This has prompted the screening of this plant for antibacterial activity. The main aims of this study were to isolate compounds from the plant's seeds and to evaluate their antibacterial effects on clinical bacterial test strains. Methods: The n-butanolic concentrate of the seed extract was subjected to thin layer chromatography (TLC) and repeated silica gel column chromatography followed by elution with various solvents. The compound was identified based on observed spectral (IR, $^1H$ NMR, $^{13}C$ NMR and high-resolution mass spectrometry) data. The well diffusion method was employed to evaluate the antibacterial activities of the isolated acyclic isoprenoid compound (final concentration: $5-15{\mu}g/mL$) on four test bacterial strains, namely, Staphylococcus aureus (MTCC 96), Bacillus cereus (MTCC 430), Escherichia coli (MTCC 1689) and Acinetobacter baumannii (MTCC 9829). Results: Extensive spectroscopic studies showed the structure of the isolated compound to be an acyclic isoprenoid ($C_{21}H_{32}O$). Moreover, the isoprenoid showed a remarkable inhibition of bacterial growth at a concentration of $15{\mu}g/mL$ compared to the two other doses tested (5 and $10{\mu}g/mL$) and to tetracycline, a commercially available antibiotic that was used as a reference drug. Conclusion: The isolation of an antimicrobial compound from Semecarpus anacardium seeds validates the use of this plant in the treatment of infections. The isolated compound found to be active in this study could be useful for the development of new antimicrobial drugs.

Antibiotic and Phytotoxic Activities of Ophiobolins from Helminthosporium Species

  • Kim, Hyun-Ju;Kim, Jin-Cheol;Kim, Byung-Sup;Kim, Hong-Gi;Cho, Kwang-Yun
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.14-20
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    • 1999
  • Twenty isolates of Helminthosporium species were obtained from various grass plants and tested for controlling efficacy on the development of plant diseases. An isolate of Helminthosporium sp. TP-4 was chosen and six antibiotic substances were purified from cultures of the fungus by repeated silica gel column chromatography and preparative thin-layer chromatography. They were identified as ophiobolin a, 6-epiophiobolin A, 3-anhydroophiobolin A, 3-anhydro-6-epiophiobolin A, iphiobolin B, and iphiobolin I mainly by mass spectrometry and nuclear magnetic resonance spectrometry. Ophiobolins inhibited the growth of a grampositive bacterium Streptomyces griseus, but were not active against gram-negative bacteria. They also showed an antifungal activity. In in vivo tests, iphiobolin B exhibited potent controlling activities against rice blast, tomato late blight, and wheat leaf rust with control values more than 90% and 70% at concentration of $500\mu\textrm{m}$/ml and 100 ${\mu}{\textrm}{m}$/ml. Ophiobolin A and 6-epiophiobolin A controlled the development of wheat leaf rust more than 80% at concentrations of 100 /ml and $500\mu\textrm{m}$/ml respectively. 3-Anhydro-6-epiophiobolin A was not active against any plant disease. On the other hand, the A-series ophiobolins other than 3-anhydroophiobolin A showed stronger phytotoxic activity in a leaf-wounding assay using 8 plant species than those of 3-anhydroophiobolin A, ophiobolin B, and ophiobolin I. The results indicate that there is little correlation between antifungal activity and phytotoxicity of ophiobolins.

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Micromachinng and Fabrication of Thin Filmes for MEMS-infrarad Detectors

  • Hoang, Geun-Chang;Yom, Snag-Seop;Park, Heung-Woo;Park, Yun-Kwon;Ju, Byeong-Kwon;Oh, Young-Jei;Lee, Jong-Hoon;Moonkyo Chung;Suh, Sang-Hee
    • The Korean Journal of Ceramics
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    • v.7 no.1
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    • pp.36-40
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    • 2001
  • In order to fabricate uncooled IR sensors for pyroelectric applications, multilayered thin films of Pt/PbTiO$_3$/Pt/Ti/Si$_3$N$_4$/SiO$_2$/Si and thermally isolating membrane structures of square-shaped/cantilevers-shaped microstructures were prepared. Cavity was also fabricated via direct silicon wafer bonding and etching technique. Metallic Pt layer was deposited by ion beam sputtering while PbTiO$_3$ thin films were prepared by sol-gel technique. Micromachining technology was used to fabricate microstructured-membrane detectors. In order to avoid a difficulty of etching active layers, silicon-nitride membrane structure was fabricated through the direct bonding and etching of the silicon wafer. Although multilayered thin film deposition and device fabrications were processed independently, these could b integrated to make IR micro-sensor devices.

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Analysis of Polybrominated Diphenyl Ethers (PBDEs) in Sediments from the Southeastern Coastal Areas of Korea

  • Choi, Hee-Gu;Kim, Sang-Soo;Jeong, Seung-Ryul;Lee, Pil-Yong;Moon, Hyo-Bang
    • Journal of the korean society of oceanography
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    • v.36 no.4
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    • pp.101-108
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    • 2001
  • Polybrominated diphenylether (PBDEs) congeners, 2,4,4'-TrBDE, 2,2',4,4'-TeBDE, 2,2',4,4',5-PeBDE, 2,2',4,4',5,5'-HXBDE, and 2,2',4,4',5,6-HxBDE, were measured in sediments from 52 stations in the southeastern coastal areas of Korea. Sediment samples were analyzed using gas chromatography coupled to mass spectrometer detector (GC/MSD) with positive electron impact (PEI) mode. New analytical methodology for PBDEs by the isotope dilution method was established using a multi-layer silica gel column chromatography. Total PBDEs levels in sediments for Pohang coast ranged from 1,1 to 33.8 ng/g dry weight (mean 5.3 ng/g dry), from 1.6 to 36.4 ng/g dry weight (mean 5.7 ng/g dry) for Ulsan coast, from 0.8 to 20.3 ng/g dry weight (mean 4.9 ng/g dry) for Busan coast, and from 0.8 to 10.3 ng/g dry weight (mean 4.4 ng/g dry) for Jinhae Bay. PBDEs contamination in surface sediments from Korean southeastern coasts was relatively moderate in comparison to that of other marine environment in the world. The predominant PBDE congeners were 2,2',4,4'-TeBDE and 2,2',4,4',5-PeBDE.

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Molecular Characterization of the α-Galactosidase SCO0284 from Streptomyces coelicolor A3(2), a Family 27 Glycosyl Hydrolase

  • Temuujin, Uyangaa;Park, Jae Seon;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1650-1656
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    • 2016
  • The SCO0284 gene of Streptomyces coelicolor A3(2) is predicted to encode an α-galactosidase (680 amino acids) belonging to glycoside hydrolase family 27. In this study, the SCO0284 coding region was cloned and overexpressed in Streptomyces lividans TK24. The mature form of SCO0284 (641 amino acids, 68 kDa) was purified from culture broth by gel filtration chromatography, with 83.3-fold purification and a yield of 11.2%. Purified SCO0284 showed strong activity against p-nitrophenyl-α-D-galactopyranoside, melibiose, raffinose, and stachyose, and no activity toward lactose, agar (galactan), and neoagarooligosaccharides, indicating that it is an α-galactosidase. Optimal enzyme activity was observed at 40℃ and pH 7.0. The addition of metal ions or EDTA did not affect the enzyme activity, indicating that no metal cofactor is required. The kinetic parameters Vmax and Km for p-nitrophenyl-α-D-galactopyranoside were 1.6 mg/ml (0.0053 M) and 71.4 U/mg, respectively. Thin-layer chromatography and mass spectrometry analysis of the hydrolyzed products of melibiose, raffinose, and stachyose showed perfect matches with the masses of the sodium adducts of the hydrolyzed products, galactose (M+Na, 203), melibiose (M+Na, 365), and raffinose (M+Na, 527), respectively, indicating that it specifically cleaves the α-1,6-glycosidic bond of the substrate, releasing the terminal D-galactose.

Synthesis and Electromagnetic Wave Absorbing Property of BaTiO3@Fe Nanofibers with Core-Shell Structure (코어-쉘 구조를 갖는 BaTiO3@Fe 나노섬유의 합성 및 전자파 흡수 특성)

  • Lee, Young-In;Jang, Dae-Hwan;Sung, Ki-Hoon;Lee, Kyuman;Choa, Yong-Ho
    • Journal of Powder Materials
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    • v.23 no.1
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    • pp.38-42
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    • 2016
  • $BaTiO_3$-coated Fe nanofibers are synthesized via a three-step process. ${\alpha}-Fe_2O_3$ nanofibers with an average diameter of approximately 200 nm are first prepared using an electrospinning process followed by a calcination step. The $BaTiO_3$ coating layer on the nanofiber is formed by a sol-gel process, and a thermal reduction process is then applied to the core-shell nanofiber to selectively reduce the ${\alpha}-Fe_2O_3$ to Fe. The thickness of the $BaTiO_3$ shell is controlled by varying the reaction time. To evaluate the electromagnetic (EM) wave-absorbing abilities of the $BaTiO_3@Fe$ nanofiber, epoxy-based composites containing the nanofibers are fabricated. The composites show excellent EM wave absorption properties where the power loss increases to the high frequency region without any degradation. Our results demonstrate that the $BaTiO_3@Fe$ nanofibers obtained in this work are attractive candidates for electromagnetic wave absorption applications.