• Title/Summary/Keyword: Ni two-step reaction

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Characterization of Biochemical Properties of Feline Foamy Virus Integrase

  • Lee, Dong-Hyun;Hyun, U-Sok;Kim, Ji-Ye;Shin, Cha-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.6
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    • pp.968-973
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    • 2010
  • In order to study its biochemical properties, the integrase (IN) protein of feline foamy virus (FFV) was overexpressed in Escherichia coli, purified by two-step chromatography, (Talon column and heparin column), and characterized in biochemical aspects. For the three enzymatic reactions of the 3'-processing, strand transfer, and disintegration activities, the $Mn^{2+}$ ion was essentially required as a cofactor. Interestingly, $Co^{2+}$ and $Zn^{2+}$ ions were found to act as effective cofactors, whereas other transition elements such as $Ni^{2+}$, $Cu^{2+}$, $La^{3+}$, $Y^{3+}$, $Cd^{2+}$, $Li^{1+}$, $Ba^{2+}$, $Sr^{2+}$, and $V^{3+}$ were not. Regarding the substrate specificity, FFV IN has low substrate specificities as it cleaved in a significant level prototype foamy virus (PFV) U5 LTR substrate as well as FFV U5 LTR substrate, whereas PFV IN did not. Finally, the 3'-processing activity was observed in high concentrations of several solvents such as CHAPS, glycerol, Tween 20, and Triton X-100, which are generally used for dissolution of chemicals in inhibitor screening. Therefore, in this first report showing its biochemical properties, FFV IN is proposed to have low specificities on the use of cofactor and substrate for enzymatic reaction as compared with other retroviral INs.

Research of Diffusion Bonding of Tungsten/Copper and Their Properties under High Heat Flux

  • Li, Jun;Yang, Jianfeng
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2011.05a
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    • pp.14-14
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    • 2011
  • W (tungsten)-alloys will be the most promising plasma facing armor materials in highly loaded plasma interactive components of the next step fusion reactors due to its high melting point, high sputtering resistance and low deuterium/tritium retention. The bonding technology of tungsten to Cu alloy was one of the key issues. In this paper, W/CuCrZr diffusion bonding has been performed successfully by inserting pure metal interlay. The joint microstructure, interfacial elements migration and phase composition were analyzed by SEM, EDS, XRD, and the joint shear strength and micro-hardness were investigated. The mock-ups were fabricated successfully with diffusion bonding and the cladding technology respectively, and the high heat flux test and thermal fatigue test were carried out under actively cooling condition. When Ni foil was used for the bonding of tungsten to CuCrZr, two reaction layers, Ni4W and Ni(W) layer, appeared between the tungsten and Ni interlayer with the optimized condition. Even though Ni4W is hard and brittle, and the strength of the joint was oppositely increased (217 MPa) due primarily to extremely small thicknesses (2~3 ${\mu}m$). When Ti foil was selected as the interlayer, the Ti foil diffused quickly with Cu and was transformed into liquid phase at $1,000^{\circ}C$. Almost all of the liquid was extruded out of the interface zone under bonding pressure, and an extremely thin residual layer (1~2 ${\mu}m$) of the liquid phase was retained between the tungsten and CuCrZr, which shear strength exceeded 160 MPa. When Ni/Ti/Ni multiple interlayers were used for bonding of tungsten to CuCrZr, a large number of intermetallic compound ($Ni_4W/NiTi_2/NiTi/Ni_3T$) were formed for the interdiffusion among W, Ni and Ti. Therefore, the shear strength of the joint was low and just about 85 MPa. The residual stresses in the clad samples with flat, arc, rectangle and trapezoid interface were estimated by Finite Element Analysis. The simulation results show that the flat clad sample was subjected maximum residual stress at the edge of the interface, which could be cracked at the edge and propagated along the interface. As for the rectangle and trapezoid interface, the residual stresses of the interface were lower than that of the flat interface, and the interface of the arc clad sample have lowest residual stress and all of the residual stress with arc interface were divided into different grooved zones, so the probabilities of cracking and propagation were lower than other interfaces. The residual stresses of the mock-ups under high heat flux of 10 $MW/m^2$ were estimated by Finite Element Analysis. The tungsten of the flat interfaces was subjected to tensile stresses (positive $S_x$), and the CuCrZr was subjected to compressive stresses (negative $S_x$). If the interface have a little microcrack, the tungsten of joint was more liable to propagate than the CuCrZr due to the brittle of the tungsten. However, when the flat interface was substituted by arc interfaces, the periodical residual stresses in the joining region were either released or formed a stress field prohibiting the growth or nucleation of the interfacial cracks. Thermal fatigue tests were performed on the mock-ups of flat and arc interface under the heat flux of 10 $MW/m^2$ with the cooling water velocity of 10 m/s. After thermal cycle experiments, a large number of microcracks appeared at the tungsten substrate due to large radial tensile stress on the flat mock-up. The defects would largely affect the heat transfer capability and the structure reliability of the mock-up. As for the arc mock-up, even though some microcracks were found at the interface of the regions, all microcracks with arc interface were divided into different arc-grooved zones, so the propagation of microcracks is difficult.

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Purification and Characterization of 2,3-Dihydroxybiphenyl 1,2- Dioxygenase from Comamonas sp.

  • Lee Na Ri;Kwon Dae Young;Min Kyung Hee
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2001.11a
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    • pp.16-25
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    • 2001
  • A genomic library of biphenyl-degrading strain Comamonas sp. SMN4 was constructed by using the cosmid vector pWE15 and introduced into Escherichia coli. Of 1,000 recombinant clones tested, two clones that expressed 2,3-dihydroxybiphenyl 1,2-dioxygenase activity were found (named pNB 1 and pNB2). From pNB1 clone, subclone pNA210, demonstrated 2,3-dihydroxybiphenyl 1,2-dioxygenase activity, is isolated. 2,3-Dihydroxybiphenyl 1,2-dioxygenase (23DBDO, BphC) is an extradiol-type dioxygenase that involved in third step of biphenyl degradation pathway. The nucleotide sequence of the Comamonas sp. SMN4 gene bphC, which encodes 23DBDO, was cloned into a plasmid pQE30. The His-tagged 23DBDO produced by a recombinant Escherichia coli, SG 13009 (pREP4)(pNPC), and purified with a Ni-nitrilotriacetic acid resin affinity column using the His-bind Qiagen system. The His-tagged 23DBDO construction was active. SDS-PAGE analysis of the purified active 23DBDO gave a single band of 32 kDa; this is in agreement with the size of the bphC coding region. The 23DBDO exhibited maximum activity at pH 9.0. The CD data for the pHs, showed that this enzyme had a typical a-helical folding structures at neutral pHs ranged from pH 4.5 to pH 9.0. This structure maintained up to pH 10.5. However, this high stable folding strucure was converted to unfolded structure in acidic region (pH 2.5) or in high pH (pH 12.0). The result of CD spectra observed with pH effects on 23DBDO activity, suggested that charge transition by pH change have affected change of conformational structure for 23DBDO catalytic reaction. The $K_m$ for 2,3-dihydroxybiphenyl, 3-metylcatechol, 4-methylcatechol and catechol was 11.7 $\mu$M, 24 $\mu$M, 50 mM and 625 $\mu$M.

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Electrosynthesis of n-PFOSF with Potential Operation (정전위법에 의한 n-PFOSF 합성)

  • Lee, Jongil;Tae, Beomseok
    • Applied Chemistry for Engineering
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    • v.7 no.3
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    • pp.473-480
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    • 1996
  • Synthesis of n-perfluorooctanesulfonyl fluoride(n-PFOSF), which is valuable precursor perfluoro-chemicals, was studied by electrochemical fluorination(ECF). Of prime concern was to investigate the cyclic voltamograms of Ni electrode in anhydrous hydrogen fluoride(AHF) with and without the reactants and to measure fluorine evolution potential. In a batch cell, chronoampherometric electrolysis and various chemical analysis such as GC, GC/MS and IR were used to understand the amphere change of electrode and the reaction paths. Fluorine equilibrium potential was found to be about 2.8V(vs. $Cu/CuF_2$) from the cyclic voltammograms and decay curves of anode potential in AHF. In batch processes, the ECF proceeded in two distinguished steps. The first step proceeded electrochemically and the second one chemically. Under 7V(vs. $Cu/CuF_2$), amount of crude products was proportional to the applied anode potential. Above 7V(vs. $Cu/CuF_2$), it had a hundred percentage with weight ratio of reactants and productivity of PFOSF was almost constant.

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Interfacial Reaction and Joint Strength of the Sn-58Bi Solder Paste with ENIG Surface Finished Substrate (Sn-58Bi 솔더 페이스트와 ENIG 표면 처리된 기판 접합부의 계면 반응 및 접합강도)

  • Shin, Hyun-Pil;Ahn, Byung-Wook;Ahn, Jee-Hyuk;Lee, Jong-Gun;Kim, Kwang-Seok;Kim, Duk-Hyun;Jung, Seung-Boo
    • Journal of Welding and Joining
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    • v.30 no.5
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    • pp.64-69
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    • 2012
  • Sn-Bi eutectic alloy has been widely used as one of the key solder materials for step soldering at low temperature. The Sn-58Bi solder paste containing chloride flux was adopted to compare with that using the chloride-free flux. The paste was applied on the electroless nickel-immersion gold (ENIG) surface finish by stencil printing, and the reflow process was then performed at $170^{\circ}C$ for 10 min. After reflow, the solder joints were aged at $125^{\circ}C$ for 100, 200, 300, 500 and 1000 h in an oven. The interfacial microstructures were obtained by using scanning electron microscopy (SEM), and the composition of intermetallic compounds (IMCs) was analyzed using energy dispersive spectrometer (EDS). Two different IMC layers, consisting of $Ni_3Sn_4$ and relatively very thin Sn-Bi-Ni-Au were formed at the solder/surface finish interface, and their thickness increased with increasing aging time. The wettability of solder joints was investigated by wetting balance test. The mechanical property of each aging solder joint was evaluated by the ball shear test in accordance with JEDEC standard (JESD22-B117A). The results show that the highest shear force was measured when the aging time was 100 h, and the fracture mode changed from ductile fracture to brittle fracture with increasing aging time. On the other hand, the chloride flux in the solder paste did not affect the shear force and fracture mode of the solder joints.