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Properties of a bilirubin oxidase from Penicillium sp. LAM 91-89 (Penicillium sp. LAM 91-89가 생산하는 bilirubin oxidase의 특성)

  • Yi, Dong-Heui;Lee, Dong-Ho;Kim, Jung-Bae;Lee, No-Woon
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.158-162
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    • 1993
  • A bilirubin oxidase produced by Penicillium sp. strain LAM 91-89 was purified and partially characterized. The enzyme was purified about 70 folds from culture broth by ethanol precipitation, first and second Sephadex G-200 column chromatography with overall yield of 12%. The molecular weight of the enzyme was estimated to be 53,000 dalton by SDS-PAGE. The optimum pH and temperature was 8.5 and $40^{\circ}C$, respectively. The enzyme was stable in the pH range $6{\sim}10$ and below $40^{\circ}C$. Activity of the enzyme was increased by the addition of $Mg^{2+}$ but was gretly inhibited by $Ag^+,\;Hg^{2+},\;Mn^{2+},\;Pb^{2+}$, iodoacetate, p-chloromercurobenzoic acid and sodium dodecyl sulfate. Among various substrates, bilirubin was favorably reacted and $K_m$ value for bilirubin was $6.67\;{\mu}mole$.

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Sequencing and Baculovirus-Based Expression of the Glycoprotein B2 Gene of HSV-2 (G)

  • Uh, Hong-Sun;Park, Jong-Kuk;Kang, Hyun;Kim, Soo-Young;Lee, Hyung-Hoan
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.482-490
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    • 2001
  • The gene for glycoprotein B (gB2) of HSV-2-strain G was subcloned, sequenced, recombinated into the lacZ-HcNPV, expressed in insect cells, and compared with the homologous gene of other HSV-2 strains. The ORF of the gB2 gene was 2,715 bp. The overall nucleotide sequence homology of te gB2 gene compared ith that of the two previously reported HSV-2 strains appeared to be over 98%. A recombinant virus named Baculo-gB2 protein in insect cells. The recombination was confirmed by a PCR and the expression was demonstrated by radio immunoprecipitation. Insect cells infected with the Baculo-gB2 virus synthesized and processed gB2 with approximately 120 kDa in the cells, and then secreted it into the culture media, where it reacted with a nomoclonal antibody to gB2. The gB2 polypeptide contained two main hydrophobic regions (a signal sequence from 1 to 23 amino acid residues, and a membrane anchor sequence from aa 745 to 798), eight N-glycosylation sites evenly distributed, and was rich in alanine (11.2%). Antibodies to this recombinant protein that were raised in mice recognized the viral gB2 and neutralized the infectivity of the HSV-2 in vitro. There results show that the gB2 protein was successfully porduced in insect cells and could be used to raise a protective neutralizing antibody. Accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

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Genomic Barcode-Based Analysis of Exoelectrogens in Wastewater Biofilms Grown on Anode Surfaces

  • Dolch, Kerstin;Wuske, Jessica;Gescher, Johannes
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.511-520
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    • 2016
  • The most energy-demanding step of wastewater treatment is the aeration-dependent elimination of organic carbon. Microbial fuel cells (MFCs) offer an alternative strategy in which carbon elimination is conducted by anaerobic microorganisms that transport respiratory electrons originating from carbon oxidation to an anode. Hence, chemical energy is directly transformed into electrical energy. In this study, the use and stability of barcode-containing exoelectrogenic model biofilms under non-axenic wastewater treatment conditions are described. Genomic barcodes were integrated in Shewanella oneidensis, Geobacter sulfurreducens, and G. metallireducens. These barcodes are unique for each strain and allow distinction between those cells and naturally occurring wild types as well as quantification of the amount of cells in a biofilm via multiplex qPCR. MFCs were pre-incubated with these three strains, and after 6 days the anodes were transferred into MFCs containing synthetic wastewater with 1% wastewater sludge. Over time, the system stabilized and the coulomb efficiency was constant. Overall, the initial synthetic biofilm community represented half of the anodic population at the end of the experimental timeline. The part of the community that contained a barcode was dominated by G. sulfurreducens cells (61.5%), while S. oneidensis and G. metallireducens cells comprised 10.5% and 17.9%, respectively. To the best of our knowledge, this is the first study to describe the stability of a synthetic exoelectrogenic consortium under non-axenic conditions. The observed stability offers new possibilities for the application of synthetic biofilms and synthetically engineered organisms fed with non-sterile waste streams.

Microstructure and Mechanical Properties of High Strength and Stretch-Flangeability Hot-Rolled Steels (고강도-신장플랜지성 열연강의 미세조직 및 기계적 성질)

  • Chun, Eun-Joon;Lee, Ju-Seung;Do, Hyeong-Hyeop;Kim, Seong-Ju;Park, Yong-Ho;Kang, Nam-Hyun
    • Korean Journal of Materials Research
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    • v.22 no.1
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    • pp.16-23
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    • 2012
  • Research into the development of high strength (1 GPa) and superior formability, such as total elongation (10%), and stretch-flangeability (50%) in hot-rolled steel was conducted with a thermomechanically controlled hot-rolling process. To improve the overall mechanical properties simultaneously, low-carbon steel using precipitation hardening of Ti-Nb-V multimicroalloying elements was employed. And, ideal microstructural characteristics for the realization of balanced mechanical properties were determined using SEM, EBSD, and TEM analyses. The developed steel, 0.06C-2.0Mn-0.5Cr-0.2(Ti + Nb + V), consisted of ferrite as the matrix phase and second phase of granular bainite with fine carbides (20-50 nm) in both phases. The significant factor of the microstructural characteristics that affect stretch-flangeability was found to be the microstructural homogeneity. The microstructural homogeneity, manifest in such characteristics as low localization of plastic strain and internally stored energy, was identified by grain average misorientation method, analyzed by electron backscattered diffraction (EBSD) and hardness deviation between the phases. In summar, a hot-rolled steel having a composition 0.06C-2.0Mn-0.5Cr-0.2(Ti + Nb + V) demonstrated a tensile strength of 998 MPa, a total elongation of 19%, and a hole expansion ratio of 65%. The most important factors to satisfy the mechanical property were the presence of fine carbides and the microstructural homogeneity, which provided low hardness deviation between the phases.

Genetic and Biochemical Characterization of Monokaryotic Progeny Strains of Button Mushroom (Agaricus bisporus)

  • Kwon, Hyuk Woo;Choi, Min Ah;Yun, Yeo Hong;Oh, Youn-Lee;Kong, Won-Sik;Kim, Seong Hwan
    • Mycobiology
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    • v.43 no.1
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    • pp.81-86
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    • 2015
  • To promote the selection of promising monokaryotic strains of button mushroom (Agaricus bisporus) during breeding, 61 progeny strains derived from basidiospores of two different lines of dikaryotic parental strains, ASI1038 and ASI1346, were analyzed by nucleotide sequencing of the intergenic spacer I (IGS I) region in their rDNA and by extracellular enzyme assays. Nineteen different sizes of IGS I, which ranged from 1,301 to 1,348 bp, were present among twenty ASI1346-derived progeny strains, while 15 different sizes of IGS I, which ranged from 700 to 1,347 bp, were present among twenty ASI1038-derived progeny strains. Phylogenetic analysis of the IGS sequences revealed that different clades were present in both the ASI10388- and ASI1346-derived progeny strains. Plating assays of seven kinds of extracellular enzymes (${\beta}$-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease) also revealed apparent variation in the ability to produce extracellular enzymes among the 40 tested progeny strains from both parental A. bisporus strains. Overall, this study demonstrates that characterization of IGS I regions and extracellular enzymes is useful for the assessment of the substrate-degrading ability and heterogenicity of A. bisporus monokaryotic strains.

Material and Geometrical Noninear Analysis of Reinforced Concrete Columns under Cyclic Loading (반복하중을 받는 철근콘크리트 기둥부재의 재료 및 기하적인 비선형 해석)

  • 김운학
    • Journal of the Earthquake Engineering Society of Korea
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    • v.3 no.1
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    • pp.55-66
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    • 1999
  • This paper presents an analytical prediction of the hysteresis behavior of reinforced concrete long column with rectangular section under the cyclic loading state. The mechanical characteristic of cracked concrete and reinforcing bar in concrete has been modeled, considering the bond effect between reinforcing bar and concrete, the effect of aggregate interlocking at crack surface and the stiffness degradation after the crack has taken place. The strength increase of concrete due to the lateral confining reinforcement has been also taken into account to model the confined concrete. The formulation of these models for concrete and reinforcing bar has been based on the smeared crack concept that the stress-strain relationship of reinforced concrete element would be defined using the average values. In addition to the material nonlinear properties, the algorithm for large displacement problem that may give an additional deformation has been formulated using total Lagrangian formulation. The analytically predicted behavior was compared with test result and they showed good agreement in overall behavior.

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EFFECTS OF BONE ENGAGEMENT TYPE&IMPLANT LENGTH ON STRESS DISTRIBUTION: A THREE DIMENSIONAL FINITE ELEMENT ANALYSIS (임플란트 매식조건에 따른 상, 하악골의 응력분포 양상에 대한 3차원 유한요소분석 연구)

  • Choi, Jeong-Hwa;Seo, Ki-Youl;Choi, Joo-Ho;Han, Jung-Suk
    • The Journal of Korean Academy of Prosthodontics
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    • v.37 no.5
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    • pp.687-697
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    • 1999
  • A finite element analysis has been utilized to analyze stress and strain fields and design a new configuration in orthopedics and implant dentistry. Load transfer and stress analysis at implant bone interface are important factors from treatment planning to long term success. Bone configuration and quality are different according te anatomy of expecting implantation site. The purpose of this study was to compare the stress distribution in maxilla and mandible accord-ing to implant length and bone engagement types. A three dimensional axi-symmetric implant model(Nobel Biocare, Gothenburg, Sweden) with surrounding cortical and cancellous bone were designed to analyze the effects of bone engagement and implant length on stress distribution. ANSYS 5.5 finite element program was utilized as an interpreting toot. Three cases of unicortical anchorage model with 7, 10, 13 mm length and four cases of bicortical anchorage model with 5, 7, 10 and 13 mm length were compared both maxillary and mandibular single implant situation. Within the limits of study, following conclusions were drawn. 1. There is a difference in stress distribution according to cortical and cancellous bone thickness and shape. 2. Maximum stress was shown at the top of cortical bone area regardless of bone engagement types. 3. Bicortical engagement showed less stress accumulation when compared to unicortical case overall. 4. Longer the implant future length, less the stress on cortical bone area, however there is no difference in mandibular bicortical engagement case.

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Simplified Moment-Curvature Relationship Model of Reinforced Concrete Columns Considering Confinement Effect (구속효과를 고려한 철근 콘크리트 기둥의 모멘트-곡률 관계 단순모델)

  • Kwak, Min-Kyoung;Yang, Keun-Hyeok
    • Journal of the Korea Concrete Institute
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    • v.28 no.3
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    • pp.279-288
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    • 2016
  • The present study simplified the moment-curvature relationship to straightforwardly determine the flexural behavior of reinforced concrete (RC) columns. For the idealized column section, moments and neutral axis depths at different stages(first flexural crack, yielding of tensile reinforcing bar, maximum strength, and 80% of the maximum strength at the descending branch) were derived on the basis of the equilibrium condition of forces and compatibility condition. Concrete strains at the extreme compression fiber beyond the maximum strength were determined using the stress-strain relationship of confined concrete, proposed by Kim et al. The lateral load-displacement curves converted from the simplified moment-curvature relationship of columns are well consistent with test results obtained from column specimens under various parameters. The moments and the corresponding neutral axis depth at different stages were formulated as a function of longitudinal reinforcement and transverse reinforcement indices and/or applied axial load index. Overall, curvature ductility of columns was significantly affected by the axial load level as well as concrete compressive strength and the amount of longitudinal and transverse reinforcing bars.

A Study of Fiber-Reinforced Material Models for the Mechanical Characteristics of Human Annulus Fibrosus (인체의 윤상인대의 역학적 특성 모사를 위한 섬유 강화 모델에 관한 연구)

  • Lim, Jun-Taek;Choi, Deok-Kee
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.35 no.6
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    • pp.619-628
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    • 2011
  • Human soft tissues, including muscles, ligaments, skin, and blood vessels, are an interesting subject because damage to them can be observed in everyday life. Besides the lack of available experimental data and the large deformation upon loading, the anisotropic and compressible nature of annulus fibrosus makes it more difficult to find a simple material model. A fiber-reinforced hyperelastic material model is used to determine the stress-strain curves upon uniaxial loading. The energy potential function for annulus fibrosus is composed of three different parts: matrix, fibers, and matrix-fiber interaction, which accounts for the angles between two families of fibers. In this paper, two different types of energy potential function for the matrix are considered, and are inserted into the fiber-reinforced model. The calculated results are compared with the Neo-Hookean model and experimental data, and reasonable agreement is observed overall.

Virulence Factors and Antimicrobial Susceptibility of Vibrio parahaemolyticus Isolated from the Oyster Crassostrea gigas (양식 굴(Crassostrea gigas)에서 분리된 장염비브리오균의 독소 유전자 보유 및 항균제 감수성)

  • Kim, Sukyung;An, Sera;Park, Bomi;Oh, Eun-Gyoung;Song, Ki Cheol;Kim, Jung-Wan;Yu, Hongsik
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.2
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    • pp.116-123
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    • 2016
  • This study investigated the prevalence of Vibrio parahaemolyticus in the oyster Crassostrea gigas, which is commonly consumed raw. The presence of virulence factors and the antimicrobial susceptibility of isolates were also investigated. The overall prevalence rate of V. parahaemolyticus in oysters was 37.5% (36/96) and the range of concentrations was 30-11,000 MPN/100 g. PCR-based assays indicated that 9.6% (11/115) of the isolates were positive for the thermostable direct hemolysin-related hemolysin gene (trh), while none of the isolates were positive for the thermostable direct hemolysin gene (tdh). The Multiple Antibiotics Resistance (MAR) index was measured for 16 common antimicrobial agents and 46.1% (53/115) of the isolates had a MAR index > 0.2. The MAR index ranged from 0.07 to 0.73. The highest MAR index was observed with strain s150608, isolated in June 2015, which exhibited resistance to 11 antimicrobial agents. Our results demonstrate that oysters are high-risk sources of V. parahaemolyticus, although no antimicrobial agent was being used to promote growth or to treat bacterial infections in the sampled oyster-growing areas.