• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1583-1590
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• 2006

T4 endonuclease V (T4 endo V) [EC 3. 1. 25. 1], found in bacteriophage T4, is responsible for excision repair of damaged DNA. The enzyme possesses two activities: a cyclobutane pyrimidine dimer DNA glycosylase (CPD glycosylase) and an apyrimidic/apurinic endonuclease (AP lyase). T4 denV (414 bp cDNA) encoding T4 en do V (138 amino acid) was synthesized and expressed using either an expression vector, pTriEx-4, in E. coli or a baculovirus AcNPV vector, pBacPAK8, in insect cells. The recombinant His-Tag/T4 endo V (rHis-Tag/T4 endo V) protein expressed from bacteria was purified using one-step affinity chromatography with a HiTrap Chelating HP column and used to make rabbit anti-His-Tag/T4 endo V polyclonal antibody for detection of recombinant T4 endo V (rT4 endo V) expressed in insect cells. In the meantime, the recombinant baculovirus was obtained by cotransfection of BacPAK6 viral DNA and pBP/T4 endo V in Spodoptera frugiperda (Sf21) insect cells, and used to infect Sf21 cells to overexpress T4 endo V protein. The level of rT4 endo V protein expressed in Sf21 cells was optimized by varying the virus titers and time course of infection. The optimal expression condition was set as follows; infection of the cells at a MOI of 10 and harvest at 96 h post-infection. Under these conditions, we estimated the amount of rT4 endo V produced in the baculovirus expression vector system to be 125 mg/l. The rT4 endo V was purified to homogeneity by a rapid procedure, consisting of ion-exchange, affinity, and reversed phase chromatographies, based on FPLC. The rT4 endo V positively reacted to an antiserum made against rHis-Tag/T4 endo V and showed a residual nicking activity against CPD-containing DNA caused by UV. This is the first report to have T4 endo V expressed in an insect system to exclude the toxic effect of a bacterial expression system, retaining enzymatic activity.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1665-1672
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• 2012

An 80% ethanol extract of Hizikia fusiforme was obtained and followed by successive fractionation using the organic solvents n-hexane, ethyl acetate, and n-butanol to identify the antioxidative substance. The aqueous part of the nbutanol fractionation step, showing high antioxidative activity, was subjected to reverse-phase liquid chromatography. As a result, a substance purified from a BB-2 fraction showed high antioxidative activity. The m/z 419 [M+H] molecular ion peak in the fraction was observed by the analysis of the ESI-LC/MS spectrum. By the analysis of 1H NMR (500 MHz, DMSO-$d_6$) and $^{13}C$ NMR (125 MHz, DMSO-$d_6$) spectra, a unique compound of the fraction was biochemically identified as a 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5HHMF). We also investigated the effect of 5HHMF on human gastric AGS carcinoma cells. Western blot analysis suggested that the flavone substantially increased the levels of the death receptor-associated apoptosis mediators Fas, Fas L, FADD, TRADD, and DR4 in a concentration-dependent manner. The levels of Fas, Fas L, TRADD, and DR4 in the cells treated with 5HHMF ($5{\mu}g/ml$) were approximately 26.4-, 12.8-, 6.7-, and 9.8-times higher than those of non-treated cells, respectively. Of note, the level of FADD protein in the cells exposed to 5HHMF ($1{\mu}g/ml$) increased approximately 9.6-times. In addition, the cleavage of caspase-3, -8, and -9 in cultured AGS cells treated with 5HHMF was significantly confirmed. Therefore, our results suggest that 5HHMF from H. fusiforme is involved in the induction of death receptor-associated apoptosis mediators in human gastric AGS carcinoma cells.

• Journal of Korean Society of Environmental Engineers
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• v.28 no.11
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• pp.1192-1197
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• 2006

Adsorbability of ionic and nonionic pharmaceuticals was studied using granular activated carbon(GAC). In a batch adsorption test of muticomponent solution, 500 mg/L of GAC dose removed all target compounds between 94 and 98% at initial concentration of 10 ${\mu}g/L$. Adsorption of ionic pharmaceuticals increased as pH was lowered toward to pKa, however adsorption capacity of nonionic pharmaceuticals showed insignificant variation with the changing pH. The enhanced adsorption capacity of ionic pharmaceuticals at lower pH was attributed to the corresponding increase in the molecular form of ionic pharmaceuticals with carboxylic group at low pH. In addition, decrease of pH increased hydrogen ion concentration in the bulk solution and the protons bound to the available sites on the carbon enhanced the removal of the ionic pharmaceuticals from solution. After 40 days of continuous operation, GAC column showed the removal of target compounds were removed by $93{\sim}99%$ at 15 min of EBCT mainly due to adsorption mechanism of GAC. At shorter EBCT than 15 min, breakthrough of CA, IBP and GFZ occurred earlier than the other ionic and nonionic pharmaceuticals. effect of EBCT on adsorption of nonionic pharmaceuticals was greater than ionic ones. This study showed that persitent pharmaceuticals found in drinking water treatment could be effectively controlled by adsorption in GAC process.

• Applied Biological Chemistry
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• v.26 no.2
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• pp.125-131
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• 1983

Formal net charges, bond populations, atomic orbital coefficients, energy components and conformation of dimethyl-2,2-dichlorovinylphosphate have been studied theoretically by using the CNDO/2 molecular orbital calculation method in attempt to describe the reactivity and the stability of the molecule. From the analysis of rate equation, molecular orbital calculations and identification of the hydrolysis products, 2,2-dichloroacetaldehyde and dimethylphosphoric acid, a mechanism of the hydrolysis of dimethyl-2,2-dichlorovinylphosphate(DDVP) has been proposed. The hydrolysis of DDVP proceeds through the mechanism of nucleophilic addition, typical Micheal reaction in basic media. Therefore, it appears probable that the attack by strong nucleophile, hydroxide ion occurs at the increased positive charge $C_2({\alpha})$ atom of a staggered conformation due to the inductive effect (-)I>(+)R of 2,2-dichlorovinyl, electron-attracting group. And then, the hydrolytic scission involves the $C_2({\alpha})-O_3$, ${\pi}-anti-bonding\;orbital({\pi}^*)$ in the subsequent reaction in aqueous solution.

• Polymer(Korea)
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• v.35 no.4
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• pp.363-369
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• 2011

To study the effect of incorporation of a surface crosslinking layer on a crosslinked poly (sodium acrylate) (cPSA) absorbent with ethylene glycol dimethacrylate CEGDMA), we synthesized several surface crosslinked cPSAs with EGDMA by an inverse emulsion polymerization method to delay the absorption of excess water in concrete, Liquid paraffin was used as a continuous phase. cPSA was synthesized with acrylic acid (AA) neutralized with aqueous 8 M sodium hydroxide solution as a monomer, N,N-methylene bisacrylamide (MBA) as crosslinking agent and ammonium persulfate (APS) and sodium metabisulfite (SMBS) as a redox initiator system by inverse emulsion polymerization. FTIR spectroscopy was used to characterize $Ca^{2+}$ ion interaction with cPSA and cPSA-EGDMAs. The swelling ratios of synthesized absorbents were evaluated from the absorption in deionized water, cement saturated aqueous solution and aqueous solution of calcium hydroxide (pH 12). Equilibrium swelling times for cPSA and surface crosslinked cPSA with EGDMA were 2 and 3 hrs, respectively. We also observed an increase in setting time of the cement and an increase in the compressive strength of mortar by addition of the synthesized cPSA-EGDMA.

• Korean Journal of Soil Science and Fertilizer
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• v.31 no.4
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• pp.414-419
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• 1998

The contribution of soil organic matter on the soil surface charge characteristic of paddy and upland soils weathered from granite or limestone was evaluated. The surface charge characteristics of the soils with and without soil organic matter by pre-treatment with hydrogen peroxide was determined at pH 3.5~9.0 range using the ion adsorption method. Regardless of soil organic matter removal, the soil surface negative charge increased linearly by the increase of pH with high statistical significance at all kinds of soils. Here, the differential increasement of soil surface negative charge by pH inclease, dCEC/dpH, was proposed as the parameter of pH dependency of the soil surface charge. The dCEC/dpH of soils with organic matter was in the range of 0.91~4.59, while it was dramatically decreased to the range 0.16~1.91 by the removal of organic matter. The soil surface charge derived from soil organic matter ranged from 15% to 82% to the total amount of surface charge. The magnitude of surface charge carried by 1% of soil organic matter showed considerable differences between soils from 0.22 to $5.03cmol^+\;kg^{-1}$. The effect of soil organic matte on the dCEC/dpH was higher in paddy soils with high oxalic acid extractable Fe than upland soils.

• Korean Journal of Soil Science and Fertilizer
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• v.31 no.4
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• pp.400-406
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• 1998

To investigate the effect of simulated acid rain on the change in soil chemical properties, simulated acid rain of different pH was applied to the three soils of different texture. Simulated acid rain of pH 4.0 and 6.0 did not greatly change the soil pH, while simulated acid rain of pH 2.0 decreased greatly the soil pH. Decrease in soil pH were in the order of sandy loam > loam > clay loam, while increase in exchangeable acidity was in the order of clay loam > loam > sandy loam. Amount of nutrients leached downward due to the penetration of simulated acid rain into the soil was in the order of Ca > K > Mg. Exchangeable Al was not detected when soil acidity dropped to pH 5 and exchangeable acidity increased within a range of CEC. A total 1200mm of simulated acid rain(pH 3.0) can load $12kg\;ha^{-1}$ of $H^+$ ion, $128kg\;ha^{-1}$ of sulfur, $56kg\;ha^{-1}$ of nitrogen. The acidity of simulated acid rain pH 3.0 can be neutralized by addition of $444kg\;ha^{-1}$ of slaked lime. The amount of leached bases were equivalent to 923, 1731 and $1608kg{\cdot}ha^{-1}$ in sandy loam, loam and clay loam soil respectively.

• The Korean Journal of Mycology
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• v.29 no.2
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• pp.91-98
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• 2001

The mycelial morphology during submerged cultivation of Ganoderma ludium using by air-lift fermenter system were analyzed by image processing system and the characterization of mycelial morphology were investigated. In submerged culture using medium with different ammonium phosphate concentrations, the various morphological forms of G. lucidum mycelium were observed. The filamentous forms such as non-branched long filamentous mycelium, non-branched short mycelium, branched long filamentous mycelium, branched short mycelium, entangled mycelium and clump were observed, and also, and also, the pelleted forms such as smooth pellet, rough pellet and hollow rough pellet were observed. The mycelial morphology was changed from the filamentous to the pelleted forms by addition of ammonium phosphate. The fractal dimensions of pelleted and filamentous forms were 1.05 and 1.3, respectively, while the fractal dimension of mixtures of pelleted and filamentous forms was 1.16. Therefore, the fractal dimension was found to be more effective index for the detection of the mycelial morphology and morphological change during batch cultivation. The circularity was also found to be useful for evaluating the surface growth of pelleted mycelium.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.6 no.2
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• pp.292-300
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• 1996

Hexavalent chromium($Cr^{+6}$) compounds are considered to be particularly hazardous, primarily because of the associated risk of allergic reaction and cancer. The analytic method of hexavalent chromium such as the s-diphenylcarba-zide(DPC) method and all ether previously used methods are often made uncertain due to significant interferences from organic components. This report can provide a technique for the more rapid and simple determination of total hexavalent chromium. than other currently using methods. The s-diphenylcarbazide method proposed by the U.S. National Institute for Occupational Safety and Health has low recovery rate(15.67 - 48.20%) due to interference, iron chloride and nickel chloride. A microwave oven technique has high recovery rate(about 70%) of insoluble hexavalent chromium. For the difference of ionic charges of $Cr^{+3}$-ethylenediamine tetraacetic acid(EDTA) chelate and $CrO_4{^{-2}}$, we could detect them simultaneously by ion exchanged high performance liquid chromatography. The confirmation of $Cr^{+3}$ and $Cr^{+6}$ were checked by fraction collector and flameless atomic absorption spectrometer. We observed that the small amount of hexavalent chromium is converted to trivalent chromium due to enhancement of chromium reduction by $Fe^{+3}$ or $Ni^{+2}$. As a result of this study, on the analysis of insoluble hexavalent chromium with microwave oven was used for, it may be better and more precise analysis after pretreatment by 2% NaOH-3% $Na_2CO_3$ and then analysis UV-spectrophotometer. It should be done for various studies on insoluble hexavalent chromium on the basis work environmental monitoring so called welding, painting etc.

• Korean Journal of Plant Tissue Culture
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• v.28 no.4
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• pp.221-225
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• 2001

Some factors of wounding methods, solidifying agents, origin of leaf explants, cone. of acetosyring-one, and MES affecting regeneration and transformation by Agrobacterium tumefaciens were investigated to establish an efficient transformation system of apple. Wounding by cutting the leaves merely showed the tendency of regeneration and transformation with higher efficiency compared with that of wounding by non-traumatic forcep when carrying out co-cultivation for three days after bacterial inoculation. While examining the solidifying agents of medium with the combination of agar (A)＋Gelrit $e^{ }$ (G) in g. $L^{-1}$ , the higher concentration of Gelrit $e^{ }$ increased the efficiency of regeneration. However, there was no difference in the efficiency of transformation from the treatments of 2.5 G, 3.5 A+1.2 G, and 7.0 G. The origin of leaf explants showed no difference statistically in the efficiency of regeneration and transformation, but that from the shoots of proliferation medium showed the tendency with higher efficiency. The concentration of above 0.1 mM acetosyringon had an increase in the efficiency of regeneration and transformation and the concentration of 0.15 mM had the highest efficiency of transformation in the treatment of acetosyringone with different concentration. There was no effect of MES on regeneration and transformation.ion.