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Cloning of the Cellulase Gene and Characterization of the Enzyme from a Plant Growth Promoting Rhizobacterium, Bacillus licheniformis K11 (고추역병 방제능이 있는 식물성장촉진 균주 Bacillus licheniformis K11의 cellulase 유전자의 cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.50 no.2
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    • pp.95-100
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    • 2007
  • The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.

Estimate of Regional and Broad-based Sources for PM2.5 Collected in an Industrial Area of Japan

  • Nakatsubo, Ryouhei;Tsunetomo, Daisuke;Horie, Yosuke;Hiraki, Takatoshi;Saitoh, Katsumi;Yoda, Yoshiko;Shima, Masayuki
    • Asian Journal of Atmospheric Environment
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    • v.8 no.3
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    • pp.126-139
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    • 2014
  • In order to estimate the influence of sources on $PM_{2.5}$ in the industrial area of Japan, we carried out a source analysis using chemical component data of $PM_{2.5}$. $PM_{2.5}$ samples were collected intermittently at an industrial area in Japan from July 2010 to November 2012. Water soluble ions ($Cl^-$, $NO_3{^-}$, $SO{_4}^{2-}$, $Na^+$,$NH_4{^+}$, $K^+$, $Mg^{2+}$, $Ca^{2+}$), elements (Al, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, As, Cd, Sb, Pb), and carbonaceous species (OC, EC) of the $PM_{2.5}$ (a total of 198 samples) were analyzed. Positive Matrix Factorization (PMF) model was applied to the data of those chemical components to identify the source of $PM_{2.5}$. At this observation site, nine factors were extracted. The major contributors of $PM_{2.5}$ were secondary sulfate 1, in which loading factors of $SO{_4}^{2-}$ and $NH_4{^+}$ were large (percentage source contribution: 20.9%), traffic, in which loading factors of OC (organic carbon) and EC (elemental carbon) were large (20.8%), secondary sulfate 2, in which loading factors of K and $SO{_4}^{2-}$ were large (8.0%), steel mills (7.8%), secondary chloride and nitrate (7.0%), soil (5.0%), heavy oil combustion (3.8%), sea salt (3.8%), and coal combustion (2.3%). The conditional probability function (CPF) and the potential source contribution function (PSCF) were carried out to examine the influence of a regional source and a broad-based source, respectively. CPF results supported local source influences such as steel mills, sea salt, traffic, coal combustion, and heavy oil combustion. PSCF results suggested that ships in the East China Sea, an industrial area of the east coastal region of China, and an active volcano in the Kyushu region of Japan were potential regional sources of secondary sulfate 1. Secondary sulfate 2 was affected by the burning of biomass fields and by coal combustion in Chinese urban areas such as Beijing, Hebei, and western Inner Mongolia. Source characterization using continuous data from one site showed a potential source representing fossil fuel combustion is affected both by regional and broad-based sources.

Characteristics of Trigeminal Evoked Potential and It's Pathway in the Rat (백서에서 삼차신경 유발전위의 특성과 경로 분석)

  • Kim, Se-Hyuk;Zhao, Chun-Zhi;Kwon, Oh-Kyoo;Lee, Bae-Hwan;Park, Yong-Gou;Chung, Sang-Sup
    • Journal of Korean Neurosurgical Society
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    • v.29 no.8
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    • pp.985-994
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    • 2000
  • Objective : There are some advantages of trigeminal evoked potential(TEP) recording compared to other somatosensory evoked potential(SSEP) recordings. The trigeminal sensory pathway has a pure sensory nerve branch, a broader receptive field in cerebral cortex, and a shorter pathway. Despite these advantages, there is little agreement as to what constitutes a normal response and what wave forms truly characterize the intraoperative TEP. This study presents the normative data of TEP recorded on the epidural surface of the rat with a platinum ball electrode. Materials & Methods : Under general anesthesia with urethane, the adult Sprague-Dawley male rats(300-350g) were given electrical stimulation with two stainless steel electrodes which were inserted into the subcutaneous layer of the area around whiskers. A reference electrode was positioned in the temporalis muscle ipsilateral to the recording site. Results : TEPs were recorded in the Par I area of somatosensory cortex and recorded most apparently on the point of 2mm posterior from the bregma and 6mm lateral from the midline. The typical wave form consisted of 5 peaks (N1-P1-N2-P2-N3 according to emerging order, upward negativity). Each latency to corresponding peaks was not influenced by the different intensities of stimulation, especially from 1 to 5mA. Average latencies of 5 peaks were in the following order ; 7.7, 11.1, 15, 22.3, 29.4ms. There was also no significant difference between latencies before and after administration of muscle relaxant(pancuronium). For the electrophysiological localization of recorded waves, the action potential of a single unit was recorded with glass microelectrode(filled with 2M NaCl, $3-5M{\Omega}$) in the thalamus of rat. A sharp wave was recorded in the VPM nucleus, in which the latency was shorter than that of N1. This suggests that all 5 peaks were generated by neural activities in the suprathalamic pathway. Conclusion : In terms of recording near-field potentials, our data also suggests that TEP in the rat may be superior to other SSEPs. In overall, these results may afford normative data for the studies of supratentorial lesions such as hydrocephalus or cerebral ischemia which can have an influence on near-field potentials.

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Deposition Characteristics of Water-soluble Inorganic Ions in the Iksan Ambient Air during Fall, 2004 (가을철 대기환경 중 수용성 이온성분의 침적특성)

  • Kang, Gong-Unn;Kim, Nam-Song;Jeon, Seon-Bok
    • Journal of Environmental Health Sciences
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    • v.32 no.4 s.91
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    • pp.359-372
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    • 2006
  • In order to investigate the daily deposition characteristics of water-soluble inorganic components in airborne deposit on the Iksan, deposition samples were collected using a deposition gauge from October 16 to November 1, 2004. Deposition samples were collected using two different sampling gauges, a dry gauge and a wet gauge, respectively. To get wet the bottom of wet gauge during the sampling period, the volume of $30{\sim}50ml$ distilled ionized water was added in a wet gauge before the beginning of each deposition sampling. Deposition samples were collected twice a day and analyzed for inorganic water-soluble anions ($Cl^-,\;{NO_3}^-,\;{SO_4}^{2-}$) and cations (${NH_4}^+,\;Na^+,\;K^+,\;Mg^{2+},\;Ca^{2+}$) using ion chromatography. Qualify control and quality assurance of analytical data were checked by the data obtained from reinjection of standard solution, Dionex cross check standard solutions, and random several deposition samples, and measured data was estimated to be reliable. Considering the deposition sample volume, the sampling time, the surface area of sampling container, and the ion concentration measured, the daily deposition amounts for measured ions were calculated in $mg/m^2$. The total daily deposition amounts of all measured ions for dry and wet gauge were $7.5{\pm}2.8$ and $17.7{\pm}4.2mg/m^2$, respectively. A significant increase in deposition amount during rainfall days was observed for both wet gauge and dry gauge, having no difference of deposition amount between in wet gauge and in dry gauge. The mean deposition of all ions measured in this study were higher in wet gauge than in dry gauge because of the surface difference of the sampling container, especially for ${NH_4}^+\;and\;{SO_4}^{2-}$. The mean deposition amounts of ${NH_4}^+\;and\;{SO_4}^{2-}$ in wet gauge were found to be about 10 times and 3 times higher than those in dry gauge, while the rest of the chemical species were equal or a little higher in wet gauge than in dry gauge. Dominant species in dry gauge were ${NO_3}^-\;and\;Ca^{2+}$, accounting for 21% and 28% of the total ion deposition, whereas those in wet gauge were ${SO_4}^{2-}\;and\;{NH_4}^+$, accounting for 19% and 41% of the total ion deposition, respectively.

Usefulness of Chlorine Dioxide to Airborne Bacteria at a Hospital Using Biological Information (생물학적 정보를 활용한 병원에서 존재하는 공기중 부유 세균에 대한 이산화염소의 유용성)

  • Jung, Suk-Yul
    • Journal of Internet of Things and Convergence
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    • v.6 no.2
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    • pp.19-24
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    • 2020
  • In the present study, using biological information of bacteria and biochemical information of chlorine dioxide gas, Gram-positive bacteria, e.g., Alloiococcus otitis, Erysipelothrix rhusiopathiae, Staphylococcus caprae, Staphylococcus lentus, and gram-negative bacteria, e.g., Acinetobacter baumannii complex, Aeromonas salmonicida, Brucella melitensis, Oligella ureolytica were used whether a plastic kit to release ClO2 gas could inhibit their growth. Overall, chlorine dioxide gas showed about 99% inhibition of bacterial growth, with less than 10 CFU. However, it was found that Gram positive Alloiococcus otitis and Gram negative Aeromonas salmonicida had more than about 50 CFU. When comparing the results of experiments with several bacteria, it suggested that the concentration of chlorine dioxide gas would be at least 10 ppm to 400 ppm for the bacterial inhibition. The results of this study could be used as basic data to evaluate the clinical usefulness of chlorine dioxide gas. If this study helps with prior knowledge to help clinicians to recognize and prevent the presence of micro-organisms that cause infections in hospitals, it would be helpful for activities such as patient care as a convergence field. In the future, it is considered that the research results will be the basis for rapidly inhibiting the microbes infected with patients by utilizing data of the information of the microbes that are inhibited for chlorine dioxide gas.

Fluorescence Anisotropy Study on the Effect of Phellodendri Cortex's Berberine on Regulation of the Function of DNA (황백(黃柏)의 berberine이 DNA의 기능조절에 미치는 영향에 관한 형광이방성 연구)

  • Lee, Seong Kyung;Han, Hyo Sang;Huh, Sung Ho
    • The Korea Journal of Herbology
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    • v.33 no.5
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    • pp.105-110
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    • 2018
  • Objectives : We tried to observe the fluorescence anisotropy and intensity of ethidium ion in the intercalating binding interaction between DNA and ethidium ions in the presence of berberine, and then tried to explain the effect of berberine on the intercalating interaction of ethidium ion with DNA. Methods : DNA(calf thymus DNA), berberine and ethidium bromide(EtBr) were purchased from Sigma-Aldrich Co. Proper amount of each compound was dissolved in 20 mM sodium phosphate buffer(pH 7.0) containing 100 mM of NaCl to prepare stock solutions. Collections of the fluorescence anisotropy and intensity data were performed on JASCO FP-8300 spectrofluorometer equipped with a polarizer and a Peltier temperature controller. The excitation of ethidium ion was done at 550 nm and the emission data were collected at 600 nm. For Stern-Volmer plot, the fluorescence data were collected at $18^{\circ}C$ and $30^{\circ}C$. Results : According to the results of this research, the weak competitive binding pattern between ethidium ion and berberine appeared in binding with DNA at low ratio of DNA to ethidium ion. But at high ratio of DNA to ethidium ion, this weak competition disappeared. Instead, berberine might bind to DNA by intercalating way. In other words, berberine could de-intercalate ethidium ion from DNA at low concentration of DNA relative to ethidium ion, but could not at high concentration of DNA relative to ethidium ion. In addition, the mechanism of fluorescence quenching of ethidium ion could also proceed differently, depending on the ratio of the amount of DNA to that of ethidium ion. Conclusions : The effect of berberine on the DNA-ethidium ion intercalating interaction could work differently, depending on the relative ratio of the amount of DNA to that of ethidium ion. This study also showed that fluorescence anisotropy analysis is very useful method to obtain detailed information for investigation of the complex binding interactions. In order to fully understand the mechanism of action of the pharmacological effect by berberine, studies on the effect of berberine on the action of proteins such as various enzymes closely related to berberine-induced medicinal effects should be continued.

Cytotoxicity of a Novel Biphenolic Compound, Bis(2-hydroxy-3-tert-butyl-5-methylphenyl)methane against Human Tumor Cells In vitro

  • Choi, Sang-Un;Kim, Kwang-Hee;Kim, Nam-Young;Choi, Eun-Jung;Lee, Chong-Ock;Son, Kwang-Hee;Kim, Sung-Uk;Bok, Song-Hae;Kim, Young-Kook
    • Archives of Pharmacal Research
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    • v.19 no.4
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    • pp.286-291
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    • 1996
  • Phenolic compounds are prevalent as toxins or environmental pollutants, but they are also widely used as drugs for various purpose including anticancer agent. A novel biphenolic compound, bis(2-hydroxy-3-tert-butyl-5-methylphenyl)methane (GERI-BPO02-A) was isolated from the fermentation broth of Aspergillus fumigatus F93 previously, and it has revealed cytotoxicity against human solid tumor cells. Its effective doses that cause 50% inhibition of cell growth in vitro against non-small cell lung cancer cell A549, ovarian cancer cell SK-OV-3, skin cancer cell SK-MEL-2 and central nerve system cancer cell XF498 were 8.24, 10.60, 8.83, $9.85\mug/ml$ respectively. GERI-BPO02-A has also revealed cytotoxicity against P-glycoproteinexpressed human colon cancer cell HCT15 and its multidrug-resistant subline HCT15/CL02, and its cytotoxicity was not affected by P-glycoprotein. We have also tested cytotoxicities of structurally related compounds of GERI-BPO02-A such as diphenylmethane, 1,1-bis(3,4dimethylphenyl)ethane, 2,2-diphenylpropane, 2-benzylpyridine, 3-benzylpyridine, $4,4^I-di-tert-butylphenyl$, bibenzyl, $2,2^I-dimethylbibenzyl$, cis-stilbene, trans-stilbene, 3-tert-butyl-4-hydroxy-5-methylphenyisulfide, sulfadiazine and sulfisomidine for studying of structure and activity relationship, and from these data we could suppose that hydroxyl group of GERI-BPO02A conducted important role in its cytotoxicity.

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Design of the Artificial Antenna System in Photosynthesis

  • Tamiaki, Hitoshi;Yagai, Shiki
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.66-69
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    • 2002
  • Zinc chlorin 1 possessing tertiary 3$^1$_hydroxy and 13$^1$-oxo groups was synthesized as a model for the antenna chlorophylls of photosynthetic green bacteria. Self-aggregation of 1 in nonpolar organic solvents was examined and compared to 2 and 3 possessing a secondary and primary 3$^1$_hydroxy group, respectively. Zinc chlorin 1 self-aggregated in I%(v/v) CH$_2$Cl$_2$-hexane to form oligomers and showed a red-shifted Qy maximum at 704 nm compared to the monomer (648 nm in CH$_2$CI2$_2$). This red-shift is larger than that of 3$^1$S-2 (648 to 697 nm) and comparable to that of3$^1$R-2 (648 to 705 nm), but smaller than that of 1 (648 to 740 nm), indicating that while a single 3$^1$-methyl group (primary to secondary OH) suppressed tight and/or extended aggregation, the additional 3$^1$-methyl group (secondary to tertiary OH) did not further suppress aggregation. The relative stability of the aggregates was in the order 3> 3$^1$R-2∼ 1 > 3$^1$S-2 as determined by visible spectral analyses. Molecular modeling calculations on oligomers of zinc chlorins 1, 3$^1$ R-2 and 3 gave similar well-ordered energy-minimized structures, while 3 stacked more tightly than 3$^1$ R- 2 and 1. In contrast, 3$^1$S-2 gave a relatively disordered (twisted) structure. The calculated oligomeric structures could explain the visible spectral data of 1-3 in nonpolar organic solvents. Moreover, self- aggregation of synthetic zinc 13$^1$_oxo-hlorins 4-6 possessing a 2-hydroxyethyl, 3-hydroxypropyl and 3- hydroxy-I-propenyl group at the 3-position in nonpolar organic solvents was discussed.

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Isolation and Characterization of Antilisterial Lactic Acid Bacteria from Kimchi

  • Kim, Jo-Min;Kim, Ki-Hwan;Kim, Song-Yi;Park, Young-Seo;Seo, Min-Jae;Yoon, Sung-Sik
    • Food Science and Biotechnology
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    • v.14 no.4
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    • pp.503-508
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    • 2005
  • Screening for antilisterial activity was performed in about three thousand isolates of lactic acid bacteria (LAB) from Chinese cabbage kimchi, and finally based on the relatively stronger antilisterial activities eight bacterial strains were selected. The bacteria were further characterized in terms of their tolerance to artificial gastric juice, pH 2.5, bile salts (0.3% oxgall), and to the different NaCl concentrations. Of the isolates, YK005 was especially investigated for its physiological characteristics due to its inhibitory activity against gram-positive Listeria monocytogenes as well as gram-negative Escherichia coli O157:H7, as they have been constantly reported to be resistant against bacteriocins produced by a number of LAB strains. YK005 was found to be rod-shaped, $3.8\;{\mu}m$ long ${\times}\;0.5\;{\mu}m$ wide, non-sporeforming, non-motile, catalase-negative, and produced gas from glucose (heterolactic). Based on the biochemical data obtained by API 50 CHL medium, the isolate was tentatively identified as Lactobacillus brevis. To validate the result obtained by the biochemical identification, rRNA-based PCR experiments using a pair of species-specific primers for L. brevis were conducted and a single band of 1400 bp was observed, which strongly indicated that YK005 belongs to L. brevis. The LAB isolates are potentially exploited as human probiotic organisms and are employed to control some food-borne pathogens like L. monocytogenes.

Portable Amperometric Perchlorate Selective Sensors with Microhole Array-water/organic Gel Interfaces

  • Lee, Sang Hyuk;Kim, Hyungi;Girault, Hubert H.;Lee, Hye Jin
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2577-2582
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    • 2013
  • A novel stick-shaped portable sensing device featuring a microhole array interface between the polyvinylchloride-2-nitrophenyloctylether (PVC-NPOE) gel and water phase was developed for in-situ sensing of perchlorate ions in real water samples. Perchlorate sensitive sensing responses were obtained based on measuring the current changes with respect to the assisted transfer reaction of perchlorate ions by a perchlorate selective ligand namely, bis(dibenzoylmethanato)Ni(II) (Ni(DBM)2) across the polarized microhole array interface. Cyclic voltammetry was used to characterize the assisted transfer reaction of perchlorate ions by the $Ni(DBM)_2$ ligand when using the portable sensing device. The current response for the transfer of perchlorate anions by $Ni(DBM)_2$ across the micro-water/gel interface linearly increased as a function of the perchlorate ion concentration. The technique of differential pulse stripping voltammetry was also utilized to improve the sensitivity of the perchlorate anion detection down to 10 ppb. This was acquired by preconcentrating perchlorate anions in the gel layer by means of holding the ion transfer potential at 0 mV (vs. Ag/AgCl) for 30 s followed by stripping the complexed perchlorate ion with the ligand. The effect of various potential interfering anions on the perchlorate sensor was also investigated and showed an excellent selectivity over $Br^-$, $NO_2{^-}$, $NO_3{^-}$, $CO{_3}^{2^-}$, $CH_3COO^-$ and $SO{_4}^{2^-}$ ions. As a final demonstration, some regional water samples from the Sincheon river in Daegu city were analyzed and the data was verified with that of ion chromatography (IC) analysis from one of the Korean-certified water quality evaluation centers.