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Molecular Cloning and Expression of Bacillus stearothermophilus $\beta$-D-Xylosidase Gene in E. coli (Bacillus stearothermophilus $\beta$-D-Xylosidase 유전자의 크로닝 및 Escherichia coli에서의 발현)

  • 오세욱;박성수;최용진;박영인
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.136-142
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    • 1992
  • Bacillus stearothemophilus isolated from soil was identified to express multiple extracellular xylanases. Two HindIII restriction fragments of 5.4 and 6.4 kb from B. stearothermophilus genomic DNA were cloned into pBR322 to obtain recombinant plasmids pMG0l and pMG02, respectively, which enabled E. coli HBlOl cells to produce $\beta$-D-xylosidase activity. By subcloning into pUC18 and Southern blotting, the loci of the $\beta$-D-xyiosidase genes were elucidated to be on non-homologous DNA fragments of 2.2 kb from pMGOl(pMG1) and 1.0 kb from pMG02(pMG2), respectively. The two enzymes produced in E. coli cleaved xylobiose, xylotriose, xylotetrose and xylotetrose to produce xylose as a major end product. The gene on pMG1, distinct from that on pMG2 was observed to encode a bifunctional protein that displayed both P-D-xylosidase (EC.3.2.1.37) and a-L-arabinofuranosidase activities (EC.3.2.1.55).

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Corrosion Behavior and Inhibition Studies of AZ31B Magnesium Alloy With and Without Cl- in the Alkaline Electrolytes in Addition with Various Inhibitor Additives

  • Shin, Yoonji;Cho, Kyehyun
    • Corrosion Science and Technology
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    • v.18 no.6
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    • pp.243-252
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    • 2019
  • The pitting corrosion and inhibition studies of AZ31B magnesium alloy were investigated in the alkaline solution (pH12) with chloride and inhibitors. The corrosion behavior of passive film with/without Cl- in the alkaline electrolyte were conducted by polarization curve and immersion tests in the presence of various additives (inhibitors) to clarify the inhibition efficiency of pitting corrosion at higher potential region. Critical concentration of pitting corrosion for Mg alloy was evaluated with 0.005 M NaCl in 0.01 M NaOH on the anodic polarization behavior. Critical pitting of AZ31B Mg alloy in 0.01 M NaOH is a function of chlorides; Epit = - 1.36 - 0.2 log [Cl-]. When the Sodium Benzoate (SB) was only used as an inhibitor, a few metastable pits developed on the Mg surface by an immersion test despite no pitting corrosion on the polarization curve meaning that adsorption of SB on the surface is insufficient protection from pitting corrosion in the presence of chloride. The role of SB and Sodium Dodecylbenzenesulfonate (SDBS) inhibitors for the Mg alloy surface in the presence of chloride was suppressed from pitting corrosion to co-adsorb on the Mg alloy surface with strong formation of passive film preventing pitting corrosion.

The Effects of Bee Venom on iNOS, TNF-α and NF-kB in RAW 264.7 Cells (봉약침액(蜂藥鍼液)이 RAW 264.7 세포의 iNOS, TNF-α 및 NF-kB에 미치는 영향(影響 ))

  • Kim, Goon-Joong;Sim, Sung-Yong;Lee, Seong-No;Kim, Kee-Hyun
    • Journal of Pharmacopuncture
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    • v.6 no.2
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    • pp.45-56
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    • 2003
  • Objective : The purpose of this study was to investigate the effects of Bee Venom on the lipopolysaccharide(LPS), sodium nitroprusside(SNP), hydrogen peroxide$(H_2O_2)$-induced expression inducilble nitric oxide synthetase(iNOS), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) and nuclear factor kappa B(NF-kB) in RAW 264.7 cells, a murine macrophage cell line. Method : The expressions of expression iNOS and TNF-${\alpha}$ were determined by western blotting with corresponding antibodies. The expressions of expression NF-kB was assayed by EMSA method. Results : 1. The 0.5, 1 and $5{\mu}g/mg$ of bee venom on LPS-induced expression of iNOS, the $5{\mu}g/mg$ of bee venom on SNP-induced expression of iNOS and the $1{\mu}g/mg$ of bee venom on $H_2O_2$-induced expression of iNOS compared with control were inhibited significantly. 2. The 0.5, 1 and $5{\mu}g/mg$ of bee venom inhibited significantly LPS and $H_2O_2$-induced expression of TNF-${\alpha}$ compared with control, respectively. The $0.5{\mu}g/mg$ of bee venom increased significantly SNP-induced expression of TNF-${\alpha}$ compared with control. 3. The $5{\mu}g/mg$ of bee venom on LPS-induced expression of NF-kB, the $0.5{\mu}g/mg$ of bee venom on SNP-induced expression of NF-kB and the 0.5, $5{\mu}g/mg$ of bee venom on $H_2O_2$-induced expression of NF-kB were inhibited significantly compared with control, respectively.

Biodegradation of a Reactive Dye, Remazol Black B in a UASB Reactor (UASB 반응기를 이용한 반응성 염료 Remazol Black B의 분해)

  • Oh, You-Kwan;Lee, Sung-Ho;Kim, Hyo-Seob;Kim, Yu-Jin;Lee, Sang-Joon;Park, Sung-Hoon
    • KSBB Journal
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    • v.14 no.6
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    • pp.688-695
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    • 1999
  • Biodegradation of the reactive dye, Remazol Black B was investigated in an upflow anaerobic sludge blanket(UASB) reactor. Important parameters studied include dye concentration(20-60 mg/L), glucose concentration as a co-substrate(1,000-3,000 mg/L), hydraulic retention time(3-24 hr), and influent pH(6.0-8.0). Under most conditions tested, the molecules of Black B were degraded readily and completely according to HPLC chromatograms. However, the color removal efficiency based on spectroscopic measurement was always approximately 75%. This suggests that the degradation products have some color intensity corresponding to 25% of the original dye molecules. The maximum influent dye concentration which satisfies the legal discharge limit of color intensity of 400 ADMI was 13 mg/L. and the highest removal rate at this dye concentration was 104 mg/L${\cdot}$day.

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Chemical Components of Red, White and Extruded Root Ginseng (홍삼 . 백삼 및 압출성형 건조수삼의 성분특성)

  • Ha, Dae-Chul;Ryu, Gi-Hyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.2
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    • pp.247-254
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    • 2005
  • The objective of this study is to compare the chemical properties of red ginseng, white ginseng, and extruded ginseng. Six kinds of samples were prepared and examined their chemical components. The comparison among crude ash, crude lipid, and total sugar resulted insignificant difference. White ginseng had lower content of reducing sugar than those of extruded ginseng and red ginseng. Total amino acid was found relatively low in treatment A (sliced whole root and dried at 7$0^{\circ}C$). Total amino acid of treatment C (extruded dry whole root ginseng slices, moisture content 30%, barrel temperature 11$0^{\circ}C$, and screw speed 200 rpm) was higher than that of treatment B (extruded dry whole root slices, moisture content 25%, barrel temperature 11$0^{\circ}C$, and screw speed 200 rpm). Crude saponin of treatments A, B, C, D (white ginseng with skin), E (skinless white ginseng), and F (red ginseng) were 4.02, 4.77, 4.12, 3.56, 3.25, and 4.02%, respectively. Ginsenoside was contained similarly as crude saponin. The amount of ginsenoside in the treatment of A, B, C, D, E, and F was recorded respectively at 6.031, 8.108, 6.876, 7.978, 5.591, and 9.834 mg/g. A specific component in red ginseng, $R_{g3}$ was detected in treatment F. Maltol was detected in treatment Band F. Acidic polysaccharide was increased 2∼3% by extrusion process. In conclusion, extruded ginseng had similar components to those of red ginseng.

Biodegradation of petroleum hydrocarbons by bacteria with surfactant producing capability and cell surface hydrophobicity (계면활성제 생성능과 세포 표면 소수성을 가진 세균 균주들에 의한 석유탄화수소의 생분해)

  • Kwon, Sun-Lul;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.265-272
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    • 2017
  • Some bacteria with different mechanisms for hydrocarbon degradation were isolated from oil-contaminated soils in Korea. Isolate Acinetobacter calcoaceticus SL1 showed biosurfactant- producing activity in oil-spreading test, and it exhibited a good emulsifying activity of 43.6 and 54.5% for diesel oil and n-hexane, respectively. It also has high cell surface hydrophobicity which can make it easily attaches to hydrocarbons and degrade them. It degraded 100% of 1,000 mg/L of n-octadecane and naphthalene, respectively in 3 days, 72.3% of 1,000 mg/L diesel oil in 7 days and 78.0% of 10,000 mg/L diesel oil in oil-contaminated soil during 28 days. Isolated strains Bacillus amyloliquefaciens S10 and B. subtilis GO9 can produce biosurfactant and formed 6.34 and 2.5 cm diameter of clear zones, respectively in oil-spreading test. Surface tension of their culture supernatant reduced from 74.6 to 34.4 and 33.3 mN/m, respectively during incubation, and critical micelle concentrations of culture supernatants were 2.0 and 5.9%, respectively. Consortium of A. calcoaceticus SL1 and B. amyloliquefaciens S10 degraded 77.8% of 10,000 mg/L diesel oil in 3 days, which indicated more efficient oil degradation than that by A. calcoaceticus SL1 alone. If these bacteria were applied together as a consortium to oil-contaminated sites, they may show a high removal rate of petroleum hydrocarbons.

The State of Marine Pollution in the Waters adjacent to Shipyards in Korea - 3. Evaluation of the Pollution of Heavy Metals in Offshore Surface Sediments around Major Shipyards in Summer 2010

  • Kim, Kwang-Soo
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.21 no.3
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    • pp.223-233
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    • 2015
  • In order to evaluate the pollution of heavy metals in offshore surface sediments around shipyards in Korea, surface sediment samples were collected at eleven stations around four major shipyards located in the southeastern coast of Korea in summer 2010 and nine kinds of heavy metals such as copper(Cu), zinc(Zn), cadmium(Cd), lead(Pb), chrome(Cr), arsenic(As), mercury(Hg), iron(Fe) and aluminum(Al) in sediments were analyzed. The concentrations of Cu at all sampling stations were in the range of 47.10~414.96 mg/kg and exceeded TEL(Threshold Effects Level) 20.6 mg-Cu/kg of Korean marine environmental standards for offshore sediments and ERL(Effect Range-Low) 34.0 mg-Cu/kg. The concentrations of Cu at seven stations around four shipyards were 65.18~414.96 mg/kg and exceeded PEL(Probable Effects Level) 64.4 mg-Cu/kg of Korean marine environmental standards for offshore sediments. The concentration of Cu at one station around B-shipyard was 414.96 mg/kg and exceeded ERM(Effect Range-Median) 270.0 mg-Cu/kg. The concentrations of Zn at all stations were in the range of 135.09~388.79 mg/kg which exceeded ERL 150.0 mg-Zn/kg. The concentrations of Zn at seven stations around four shipyards were 157.57~388.79 mg/kg and exceeded PEL 157.0 mg-Zn/kg. The concentration of Zn at one station around B-shipyard was 388.79 mg/kg and was approaching ERM 410.0 mg-Zn/kg. The concentrations of Cd at all stations were in the range of 0.11~0.54 mg/kg and were below TEL 0.75 mg-Cd/kg and ERL 1.2 mg-Cd/kg. The concentrations of Pb at all stations were in the range of 18.04~105.62 mg/kg. The concentrations of Pb at two stations around B-shipyard were 73.87~105.62 mg/kg which exceeded TEL 44.0 mg-Pb/kg and ERL 46.7 mg-Pb/kg, and were below PEL 119.0 mg-Pb/kg and ERM 218.0 mg-Pb/kg. The concentrations of Cr at all stations were in the range of 51.26~85.39 mg/kg. The concentration of Cr at one station around B-shipyard was 85.39 mg/kg and exceeded ERL 81.0 mg-Cr/kg. The concentrations of As at all stations were in the range of 8.70~22.15 mg/kg which exceeded ERL 8.2 mg-As/kg and were below ERM 70.0 mg-As/kg. The concentrations of As at eight stations around A-shipyard, B-shipyard and D-shipyard were 14.93~22.15 mg/kg which exceeded TEL 14.5 mg-As/kg and were below PEL 75.5 mg-As/kg. The concentrations of Hg at all stations were in the range of 0.02~0.35 mg/kg. The concentrations of Hg at three stations around A-shipyard were 0.11~0.13 mg/kg which were almost equal to TEL 0.11 mg-Hg/kg. Those at two stations around B-shipyard were 0.27~0.35 mg/kg which exceeded TEL 0.11 mg-Hg/kg and ERL 0.15 mg-Hg/kg, and were below PEL 0.62 mg-Hg/kg and ERM 0.71 mg-Hg/kg. The concentrations of Fe and Al at all stations were in the range of 2.90 3.66 % and 3.12 6.80 %, respectively. These results imply that heavy metals such as copper, zinc, lead, arsenic and mercury were likely to be transferred to marine environment from shipyards, especially from B-shipyard.

Purification and Characterization of Two Thermostable Xylanases from Paenibacillus sp. DG-22

  • Lee, Yong-Eok;Lim, Pyung-Ok
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.1014-1021
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    • 2004
  • Two thermostable xylanases, designated XynA and XynB, were purified to homogeneity from the culture supernatant of Paenibacillus sp. DG-22 by ion-exchange and gel-filtration chromatography. The molecular masses of xylanases A and B were 20 and 30 kDa, respectively, as determined by SDS-PAGE, and their isoelectric points were 9.1 and 8.9, respectively. Both enzymes had similar pH and temperature optima (pH 5.0-6.5 and $70^{\circ}C$), but their stability at various temperatures differed. Xylanase B was comparatively more stable than xylanase A at higher temperatures. Xylanases A and B differed in their $K_m$ and $V_{max}$ values. XynA had a $K_m$ of 2.0 mg/ml and a $V_{max}$ of 2,553 U/mg, whereas XynB had a K_m$ of 1.2 mg/ml and a $V_{max}$, of 754 U/mg. Both enzymes were endo-acting, as revealed by their hydrolysis product profiles on birchwood xylan, but showed different modes of action. Xylotriose was the major product of XynA activity, whereas XynB produced mainly xylobiose. These enzymes utilized small oligosaccharides such as xylotriose and xylotetraose as substrates, but did not hydrolyzed xylobiose. The amino terminal sequences of XynA and XynB were determined. Xylanase A showed high similarity with low molecular mass xylanases of family 11.

Effect of Mg Additive in the Bi1.84Pb0.34Sr1.91Ca2.03Cu3.06O10+δ(110 K phase) Superconductors (Bi1.84Pb0.34Sr1.91Ca2.03Cu3.06O10+δ(110 K 상)산화물 고온초전도체에 Mg 첨가에 따른 영향)

  • 이민수
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.16 no.6
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    • pp.522-531
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    • 2003
  • Samples with the nominal composition, B $i_{1.84}$P $b_{0.34}$S $r_{1.91}$C $a_{2.03}$C $u_{3.06}$ $O_{10+{\delta}}$ high- $T_{c}$ superconductors containing MgO as an additive were fabricated by a solid-state reaction method. Samples with MgO of 5~30 wt% each were sintered at 820~86$0^{\circ}C$ for 24 hours. The structural characteristics, critical temperature, grain size and image of mapping with respect to MgO contents were analyzed by XRD(X-Ray Diffraction), SEM(Scanning Electron Microscope) and EDS(Energy dispersive X-ray spectrometer) respectively. As MgO contents increased, intensity of MgO Peaks and ratio of Bi-2212 phase in superconductors intensified and the proportion of the phase transition from Bi-2223 to Bi-2212 was increased.