• Korean Journal of Microbiology
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• v.42 no.3
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• pp.195-198
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• 2006

Lethal toxin is a critical virulence factor of anthrax. It is composed two protein: protective antigen (PA) and lethal factor (LF). PA binds to specific cell surface receptors and, forms a membrane channel that mediates entry of LF into the cell. LF is a zinc-dependent metalloprotease, which cleaves MKKs [MAPK (mitogen-activated protein kinase) kinases] at peptide bonds very close to their N-termini. In this study, we suggest application of cell-based assays in the early phase of drug discovery, with a particular focus on the use of yeast cells. We constructed MEK1 expression system in yeast to determine LF activity and approached cell-based assay system to screen inhibitors, in which the results covering the construction of LF-substrate in yeast expression vector, expression, and LF-mediated proteolysis of substrate were described. These results could provided the basic steps in design of cell-based assay system with the high efficiency, rapidly and easy way to screening of inhibitors.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.7 no.6
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• pp.1313-1318
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• 2006

The mechanism fur the surface film formation was studied by in situ Atomic Force Microscopy (AFM) observation of a highly oriented pyrolytic graphite (HOPG) basal plane surface during cyclic voltammetry at a slow scan-rate of 0.5 mV $s^{-1}$ in 1 moi $dm^{-3}$ (M) $LiPF_6$ dissolved in a mixture of ethylene carbonate (EC) and diethyl carbonate (DEC). Decomposition of the electrolyte solution began at a potential around 2.15 V vs. $Li^+$/Li on step edges. In the potential range 0.95-0.8 V vs. $Li^+$/Li, flat areas (hill-like structures) and large swelling appeared on the surface. It is considered that these two features were formed by the intercalation of solvated lithium ions and their decomposition beneath the surface, respectively. At potentials more negative than 0.80 V vs. $Li^+$/Li, particle-like precipitates appeared on the basal plane surface. After the first cycle, the thickness of the precipitate layer was 30 nm. The precipitates were considered to be decomposition of the lithium salt ($LiPF_6$) and solvent molecules (EC and DEC), and to have an important role in suppressing further solvent decomposition on the basal plane.

• Korean Journal of Microbiology
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• v.41 no.4
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• pp.275-280
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• 2005

Anthrax is an infectious disease caused by the gram-positive bacterium, Bacillus anthracis. Anthrax toxin is a tripartite toxin comprising of protective antigen (PA), lethal factor (LF) and edema factor (EF). PA is the receptor-binding component, which facilitates the entry of LF or EF onto the cytosol. LF is a zinc-dependent metalloprotease, which is a critical virulence factor in cytotoxicity of infected animals. Therefore, it is of interest to develop its potent inhibitors for the neutralization of anthrax toxin. The first step to identify the inhibitors is the development of a rapid, sensitive, and simple assay method with a high-throughput ability. Much efforts have been concentrated on the preparation of powerful assays and on the screening of inhibitors using these system. In the present study, we have tried to construct anthrax lethal factor in yeast expression system to prepare cell-based high-throughput assay system. Here, we have shown the results covering the construction of a new vector system, subcloning of LF gene, and the expression of target gene. Our results are first trial to express LF gene in eukaryote and provide the basic steps in design of cell-based assay system.

• Korean Journal of Microbiology
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• v.55 no.2
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• pp.123-130
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• 2019

Chelatase catalyzes the insertion of divalent metal into tetrapyrrole and plays a key role in the biosynthesis of metallated tetrapyrroles, such as cobalamin, siroheme, heme, and chlorophyll. SirB is a sirohydrochlorin (SHC) chelatase that generates cobalt-SHC or iron-SHC by inserting cobalt or iron into the center of sirohydrochlorin tetrapyrrole. To provide structural insights into the metal-binding and SHC-recognition mechanisms of SirB, we determined the crystal structure of SirB from Bacillus subtilis subsp. spizizenii (bssSirB) in complex with cobalt ions. bssSirB forms a monomeric ${\alpha}/{\beta}$ structure that consists of two domains, an N-terminal domain (NTD) and a C-terminal domain (CTD). The NTD and CTD of bssSirB adopt similar structures with a four-stranded ${\beta}-sheet$ that is decorated by ${\alpha}-helices$. bssSirB presents a highly conserved cavity that is generated between the NTD and CTD and interacts with a cobalt ion on top of the cavity using two histidine residues of the NTD. Moreover, our comparative structural analysis suggests that bssSirB would accommodate an SHC molecule into the interdomain cavity. Based on these structural findings, we propose that the cavity of bssSirB functions as the active site where cobalt insertion into SHC occurs.

• Journal of the Korean Home Economics Association
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• v.39 no.1
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• pp.91-102
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• 2001

The non-starch polysaccharides in the cell wall of rice, glutinous rice, and black rice, were analyzed. They were fractionated into fractions; water-soluble, hot writer-soluble, ammonium oxalate-soluble, sodium hydroxide-soluble, potassium hydroxide-soluble, and the alkali-insoluble, according to the solvent solubility. The dietary fiber contents were 5.4% in glutinous rice, 4.2% in rice and 7.5% in black rice. The sodium hydroxide soluble fibers were abundant in each kind of rice, especially 4.01% in black rice. The water soluble fibers were 80% of dietary fiber in glutinous rice, 66% in rice, 86% in black rice. It was supposed that the content of the water soluble fibers in rice was increased by pounding. Pectic substances in water soluble fibers, hot water soluble fibers, and ammonium oxalate soluble fibers fraction, were 2.4% in glutinous rice fraction,1.59% in rice, and 1.12% in black rice. Alkali soluble fibers were considered as hemicellulose. Black rice contained 5.80% of hemicellulose, which was more than twice as much as glutinous rice(2.58%) and rice(2.22%). Alkali insoluble fibers were considered as cellulose, which showed no considerable difference. Among samples content of uronic acid in glutinous rice, rice and black rice were 0.9%, 0.66%, 1.8% respectively. Uronic acid of black rice was twice more than other samples tested. The fraction of black rice that uronic acid was extracted at most was the fraction of sodium hydroxide. Mono saccharides of the fraction was the glucose, the arabinose, the xylose.

• Journal of Life Science
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• v.27 no.10
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• pp.1111-1120
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• 2017

This study was conducted to select pepper lines that were tolerant to excessive water injury among the pepper germplasm and investigate the genetic characteristics of those lines to contribute to the breeding of pepper cultivars with stable productivity in abnormal weather. Each of the tolerant and susceptible lines went through immersion treatment, and differentially expressed genes between them were analyzed. The tolerant line showed increased expression of the CA02g26670 gene, which is involved in the CONSTANS protein pathway and regulation of flowering by day length, but it exhibited decreased expressions of CA01g21450, CA01g22480, CA01g34470, CA02g00370 and CA02g00380. The susceptible line showed increased gene expressions of CA02g09720, CA02g21290, CA03g16520, CA07g 02110, and CA12g17910, which are involved in the inhibition of proteolytic enzyme activity, DNA binding, inhibition of cell wall-degrading enzyme, and inhibition of nodulin, respectively. Meanwhile the expressions of CA02g02820, CA03g21390, CA06g17700 and CA07g18230 decreased in the susceptible line, in relation to calcium-ion binding, high temperature, synthesis of phosphocholine and cold stress, respectively. The expressions of genes related to apoptosis and peroxidase increased, while that of CA02g16990, which functions as a nucleoside transporter, decreased in both the tolerant and susceptible lines. Based on the different gene expressions between the tolerant and susceptible lines, further studies are needed on breeding abiotic stress-tolerant lines.

• The Korean Journal of Pharmacology
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• v.29 no.2
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• pp.175-182
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• 1993

As it has been reported that the depolarization-induced ACh release is modulated by activation of presynaptic $A_1-adenosine$ heteroreceptor in hippocampus and various lines of evidence indicate the adenosine effect is magnesium dependent, the present study was undertaken to delineate the role of endogenus adenosine as a modulator of hippocampal acetylcholine release in this study. Slices from the rat hippocampus were equilibrated with $[^3H]-choline$ and the release of the labelled product, $[^3H]-ACh$, was evoked by electrical stimulation(3Hz, $5\;V\;cm^{-1},$ 2ms, rectangular pulses), and the influence of various agents on the evoked tritium outflow was investigated. Adenosine, in concentrations ranging from $0.3\;to\;100\;{\mu}M$, decreased the $[^3H]-ACh$ release in a dose-dependent manner without changing the basal rate of release. $DPCPX(1{\sim}10{\mu}M)$, a selective $A_1-receptor$ antagonist, increased the $[^3H]-ACh$ release in a dose-related fashion with slight increase of basal tritium release. And the effects of adenosine were significantly inhibited by $DPCPX(2{\mu}M)$ treatment. CPCA, a specific $A_2-agonist$, in concentration ranging from $0.3\;to\;30\;{\mu}M$ decreased evoked tritium outflow with increase of basal rate of tritium release, and these effects were also abolished by $DPCPX(2{\mu}M)$ pretreatment. But, $CGS(0.1{\sim}10{\mu}M)$, a recently introduced potent $A_2-agonist$, did not alter the evoked tritium outflow. When the magnesium concentration of the medium was reduced to 0 mM, there was no change in evoked ACh release by adenosine. In contrast, increasing the magnesium concentration to 4 mM, the inhibitory effects of adenosine were significantly potentiated. These results indicate that $A_1-adenosine$ heteroreceptor is involved in ACh-release in the rat hippocampus and the inhibitory effects of adenosine mediated by $A_1-receptor$ is magnesium-dependent.

• Microbiology and Biotechnology Letters
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• v.42 no.1
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• pp.58-66
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• 2014

In the course of study for a use for non-edible parts of broccoli (Brassica oleracea L), and the development of processed food utilizing these parts, edible floret and non-edible stalk were extracted with ethanol and different organic solvent fractions were prepared. With 10 different extracts and fractions, their useful components and various biological activities, such as antioxidant, antimicrobial and anti-proliferation activity, were investigated. The stalk has more abundant water soluble carbohydrate when compared with the floret, and floret has higher hexane-soluble pigments. Analysis of total flavonoid and total polyphenol contents showed that the floret has 1.5~1.99 times higher concentrations than the stalk. Among the fractions, ethylacetate (EA) fractions have the highest amount of total flavonoid and total polyphenol. The stalk and floret possessed 9.45 and 42.01 mg-total flavonoid/g, respectively. In the antioxidation activity assay, the EA fraction of floret showed strong radical scavenging activity and reducing power, while the n-hexane fraction of the stalk exhibited nitrite scavenging activity. In the antimicrobial activity assay, the EA fraction of floret showed a strong and broad-range of antibacterial activity, irrespective of gram positive or gram negative bacteria. In a while, the hexane and EA fractions revealed anti-proliferative effects against the human colorectal cancer cell HCT-116. Strong anti-proliferative activities were found in the hexane fraction of stalk (18.4% of cell viability), and the n-butanol fraction of floret (6.9% of cell viability). Our results suggest that the further study of the characterization of active fractions and the identification of active components from different parts of broccoli are needed to develop functional foods or novel plant-derived medicines.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.1
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• pp.1-10
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• 2005

The purpose of this study was to evaluate the potential of a gamma irradiation to decompose 2,4,6-trinitrotoluene(TNT) in an aqueous solution. The decomposition reaction of TNT by gamma irradiation was a pseudo first-order kinetic over the applied initial concentrations($25{\sim}100mg/L$). The dose constant was strongly dependent on the initial TNT concentration. The removal of TNT was more efficient at pH below 3 and at pH above 11 than at neutral pH(pH 5-9). The required irradiation dose to remove over 99% of TNT was 40, 80 and 10 kGy, individually at pH 2, 7 and 13. The dose constant was increased by 1.6 fold and over 15.6 fold at pH 2 and 13, respectively, when compared with that at pH 7 When irradiation dose of 200 kGy was applied, the removal efficiencies of TOC were 91, 46 and 53% at pH 2, 7 and 13, respectively. Ammonia and nitrate were detected as the main nitrogen byproducts of TNT and glyoxalic acid and oxalic acid were detected as organic byproducts. The results showed that a gamma irradiation was an attractive method for the decomposition of TNT in an aqueous solution. However, regarding the application of high energy radiation for the TNT decomposition and mineralization, an application of an acidic pH below 3 to the solution before irradiation should be considered.

• Journal of the Korean Geotechnical Society
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• v.15 no.2
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• pp.153-164
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• 1999

The solubilization of BTEX was evaluated in aqueous surfactant solutions with and without several additives. Anionic surfactant(Sodium Dodecyl Sulfate, SDS) and nonionic surfactants (NEODOL(equation omitted)25-3 and $SOFTANOL\circledR-90$ were used as test surfactants. The effects of surfactant HLB(Hydrophile-Lipophile Balance) Number and hydrocarbon molar volume and polarity of BTEX on the MSR(Molar Solubilization Ratio), micelle-water partition coefficient of BTEX, and CMC(C,itical Micelle Concentration) were investigated. Optimizing treatment conditions applicable to enhanced solubilization was also studied by manupulating salinity or electrolyte control with additives of ethyl alcohol, hydrotrope, and electrolyte solution. The most effective surfactant for solubilization was found $SOFTANOL\circledR-90$, since HLB number of 13.6 is similar to those values of BTEX ranging between 11.4 and 12.2, which was also proved experimentally. Ethyl alchohol of 3% was the most effective additives in reducing CMC and improving solubilization among the conditions using SDS, NEODOL(equation omitted)25-3, and $SOFTANOL\circledR-90$ with three additives. The partitioning of BTEX between surfactant micelles and aqueous solutions was characterized by a mole fraction micelle-phase/aqueous phase partion coefficient, $K_m$. Values of log $K_m$. for BTEX compounds in surfactant solutions of this study range from 2.95 to 3.76(100mM SDS) and 2.95 to 3.49(117mM $SOFTANOL\circledR-90$. Log $K_m$ appears to be a linear function of log $K_{ow}$ for SDS and $SOFTANOL\circledR-90$. A knowledge of partitioning of BTEX in aqueous surfactant system can be a prerequisite for the understanding of the behavior of hydrophobic organic compounds in soil-water systems in which surfactants play a role in remediation of contaminated soil and facilitated transport.