• Title/Summary/Keyword: $Mn^{2+}$ ion

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Characterization of Xylanase from an Hybird between Aspergillus oryzae var. oryzae and Aspergillus Nidulans 514 by Nuclear Transfer (핵전이에 의한 Aspergillus oryzae var. Oryzae와 Aspergillus nidulans 514의 잡종으로부터 생산된 Xylanase의 특성)

  • Yang, Young-Ki;Moon, Myeng-Nim;Park, Hyung-Nam;Lim, Chae-Young
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.50-58
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    • 1996
  • Interspecific hybrids between Aspergillus oryzae var oryzae and A. nidulans 514 were obtained by nuclear transfer technique. Several autotrophic mutants isolated from conidiospores of the two strains were mutagenized with ultraviolet and N-methyl-N-nitrosoguanidine. Optimal conditions for formation of intergeneric hybrids were investigated. Frequencies of hybrid formation by nuclear transfer were $3{\times}10^{-5}{\sim}1{\times}10^{-5}$. From observation of genetic stability, conidial size, DNA content, and nuclear stain, it was suggested that their karyptypes are aneuploid. The hybrids showed 1.1~1.4 fold higher xylanase activities than parental strains did. The xylanase of Aspergiilus sp. TAVD514-3 was purified and some of it's enzymatc characteristics were investigated. The enzyme was purified about 85 fold with an overall yield of 17% from the culture medium by ammonium sulfate fractionation, Sephadex G-75 gel permeation chromatography, and CM-sephadex A-50 ion exchange chromatography. The purified enzyme functions optimally at pH 9.0 and 80$^{\circ}C$. The enzymatic activity was increased by the presence of $Mg^{2+}$ and $Mn^2$ ions.

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Characterization of Glycerol Dehydrogenase from Thermoanaerobacterium thermosaccharolyticum DSM 571 and GGG Motif Identification

  • Wang, Liangliang;Wang, Jiajun;Shi, Hao;Gu, Huaxiang;Zhang, Yu;Li, Xun;Wang, Fei
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1077-1086
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    • 2016
  • Glycerol dehydrogenases (GlyDHs) are essential for glycerol metabolism in vivo, catalyzing its reversible reduction to 1,3-dihydroxypropranone (DHA). The gldA gene encoding a putative GlyDH was cloned from Thermoanaerobacterium thermosaccharolyticum DSM 571 (TtGlyDH) and expressed in Escherichia coli. The presence of Mn2+ enhanced its enzymatic activity by 79.5%. Three highly conserved residues (Asp171, His254, and His271) in TtGlyDH were associated with metal ion binding. Based on an investigation of glycerol oxidation and DHA reduction, TtGlyDH showed maximum activity towards glycerol at 60℃ and pH 8.0 and towards DHA at 60℃ and pH 6.0. DHA reduction was the dominant reaction, with a lower Km(DHA) of 1.08 ± 0.13 mM and Vmax of 0.0053 ± 0.0001 mM/s, compared with glycerol oxidation, with a Km(glycerol) of 30.29 ± 3.42 mM and Vmax of 0.042 ± 0.002 mM/s. TtGlyDH had an apparent activation energy of 312.94 kJ/mol. The recombinant TtGlyDH was thermostable, maintaining 65% of its activity after a 2-h incubation at 60℃. Molecular modeling and site-directed mutagenesis analyses demonstrated that TtGlyDH had an atypical dinucleotide binding motif (GGG motif) and a basic residue Arg43, both related to dinucleotide binding.

Characteristics and Cancerostatic Activity of the Starfish Lectin (별불가사리 렉틴의 특성 및 암 세포 성장저해 효과)

  • Jeune, Kyung-Hee;Park, Chae-Soo;Park, Won-Hark;Choi, Soo-Jeong;So, Myung-Suk;Chung, See-Ryun
    • YAKHAK HOEJI
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    • v.41 no.4
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    • pp.421-432
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    • 1997
  • A new lectin was partially purified from starfish,Asterina pectinifera by means of physiological saline extraction, salt fractionation, ion exchange chromatography and hy droxyapatite chromatography, and it was named APL. The biochemical properties of the APL were characterized. In addition, its effects on lymphocyte mitogenicity and cancer cell agglutinability were tested. The APL agglutinated nonspecifically human erythrocytes and rabbit blood cells. Agglutinability was decreased to 30% of control activity below pH 5 and above pH 9 and was relatively unstable at increasing temperatures above 60$^{\circ}C$. The activity was reduced by addition of two kinds of metal ions, $Ba^{2+},\;Mn^{2+}$ and chelating agent, EDTA. APL was proved to be glycoproteins containing 9% sugars. For carbohydrate specificity, it was found that the activity of APL was inhibited by D(+)-glucosamine, D(+)-galactosamine, stachyose, N-acetyl-galactosamine and methyl-${\alpha}$-D-galactopyranoside among 35 sugars tested. In amino acid composition, the contents of acidic amino acids such as aspartic acid and glutamic acid were relatively high. This result suggest that the isoelectric point would be in a lower range. APL was found that it promotes the division of human lymphocytes. APL was proved to be a potent agglutinin for cancer cells such as HeLa, L929 and L1210 cells. Significant changes on the HeLa cell surfaces affected by APL were observed under the electron microscope.

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Studies on Degradation of Nucleic acid and Related Compounds by Microbial Enzymes (미생물 효소에 의한 핵산 및 그의 관련물질의 분해에 관한 연구)

  • Kim, Sang-Soon
    • Applied Biological Chemistry
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    • v.13 no.2
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    • pp.111-129
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    • 1970
  • As a series of studies on the nucleic acids and their related substances 210 samples were collected from 76 places such as farm soil, compost of heap, nuruk and meju to obtain microbial strains which produce 5'-phosphodiesterase. From these samples total of 758 strains were isolated by the use of dilution pour plate method. For all isolated strains primary screening of the productivity of RNA depolymerase was performed and useful strains with regard to 5'-phosphodiesterase productivities were identified. For these useful strains optimum condition, the effect of various compounds on the activity of 5'-phosphodiesterase, and the optimum condition for enzyme reaction were discussed. The quantitative of 5'-mononucleotides produced by the action of 5'-phosphodiesterase was performed using anion-exchange column chromatography and their identified was done by paper chromatography, thinlayer chromatography, ultra violet spectrophotometry, and characteristic color reaction using carbazole and schiff's reagent. (1) Penicillium citreo-viride PO 2-11 and Streptomyces aureus SOA 4-21 from soil were identified as a potent 5'-phosphodiesterase producing strains. (2) Optimum culture conditions for Penicillium citreo-viride PO 2-11 strain isolated were found to be pH 5.0 and $30^{\circ}C$, and the optimum conditions for enzyme action of 5'-phosphodiesterase were pH 4.2 and $60^{\circ}C$. Best carbon source for the production of 5'-phosphodiesterase was found to be sucrose and ammonium nitrate for nitrogen source. Addition of 0.01% corn steep liquor or yeast extract exhibited 20% increase in the amount of 5'-phosphodiesterase production compared to the control. 5'-phosphodiesterase produced by this strain was activated by $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by EDTA, citrate, $Cu^{++},\;CO^{++}$. 5'-phosphodiesterase produced 5'-mononucleotide from RNA at a rate of 65.81%, and among the 5'-mononucleotides accumulated 5'-GMP only was found to have flavorous and the strain was also found lack of 5'-AMP deaminase. Productivity of flavorous 5'-GMP was found to be 186.7mg per gram of RNA. (3) Optimum culture canditions for the isolated Streptomyces aureus SOA 4-21 strain were pH 7.0 and $28^{\circ}C$, and the optimum conditions for the action of 5'-phosphodiesterase were pH 7.3 and $50^{\circ}C$. The best carbon source for 5'-phosphodiesterase production was found to be glucose and that of nitrogen was asparagine. Addition of 0.01% yeast extract exhibited increased productivity of 5'-phosphodiesterase by 40% compared to the non-added control. 5'-phosphodiesterase produced by this strain was activated by $Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by citrate, EDTA, $Cu^{++}$. It was also found that the strain produce 5'-AMP deaminase in addition to 5'-phosphodiesterase. For this reason although decomposition rate was 63.58% the accumulation of 5'-AMP, 5'-CMP, 5'-GMP and 5'-UMP occurred by the breakdown of RNA. In the course of these reaction 5'-AMP deaminase converted 60% of 5'-AMP thus produced into 5'-IMP and flavorous 5'-mono nucleotide production was significantly increased by this strain over the above mentioned one. Production rates were found to be 171.8mg per grain of RNA for 5'-IMP and 148.2mg per gram of RNA for 5'-GMP, respectively.

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A Study of Interference Factor of Analysis Method of Hexavalent Chromium in Soil using UV/VIS Spectrometry and Application of Ion Chromatography (자외선/가시선 분광법을 이용한 토양 중 6가 크롬 분석방법의 방해요인 분석 및 이온크로마토그래피 적용에 관한 연구)

  • Noh, Hoe-Jung;Lee, Myung-Gyu;Lee, Hyeon Seok;Kim, Mira;Kim, Dongho;Kim, Tae Seung
    • Journal of Soil and Groundwater Environment
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    • v.17 no.6
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    • pp.33-42
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    • 2012
  • This study was performed to find the problems according to interference factors (organic matter, pH, Cr, Mn, Fe, clay, and etc.) when we analyzed the hexavalent chromium [Cr(VI)] in soils using UV/VIS spectrometer (US EPA 7196A), attempted to evaluate the domestic applicability of analytical method (US EPA 7199) using IC-UV/VIS spectrometer as alternative method. The recovery rate of certified reference materials was 75.0% (US EPA 7196A) and 101.4% (US EPA 7199) by the analytical methods. As the results of performing QA/QC about US EPA 7199, method detection limit (MDL) and limit of quantification (LOQ) were 0.062 mg/kg, 0.196 mg/kg, respectively. The LOQ of US EPA 7199 was lower than that of the current soil official testing method in Korea (0.5 mg/kg). Cr(VI) contents in 23 soil samples were compared by the analytical methods of EPA 7196A and 7199. Cr(VI) was detected in 13 of 23 soil samples by EPA 7196A, while EPA 7199 was not detected in any soil samples. The Cr(VI) content in 23 soil samples by EPA 7196A was not clearly correlated with Cr, Fe, Mn and clay content in the soil samples. However, the contents of Cr(VI) and organic matter of the soil samples had the highest coefficient of determination ($R^2$) of 0.80. In order to evaluate the correlation between the recovery rates of Cr(VI) and organic matter contents in the soil samples, the recovery rates of 5 soil samples added Cr(VI) standard solution were analyzed by the analytical methods. According to the results, the higher the organic matter contents in soil samples, the lower the recovery rates of Cr(VI) by US EPA 7196, while in case of US EPA 7199, the recovery rates were stable regardless of the organic matter contents.

Biochemical and Molecular Characterization of Laccases from Wild Mushrooms

  • Ro, Hyeon-Su
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.43-43
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    • 2014
  • White rot fungi have been useful source of enzymes for the degradation of environmental pollutants including polycyclic aromatic hydrocarbons (PAHs) and synthetic dyes. PAHs are widespread organic compounds present in fossil fuels and are routinely generated by incomplete fuel combustion. PAHs are some of the major toxic pollutants of water and soil environments. Synthetic dyes are major water-pollutants, which are toxic to organisms in water environments and interfere photosynthesis of water plants. Removal of PAHs and synthetic dyes has been of interests in the environmental science especially in the environmental microbiology. Mushrooms are fungal groups that function as primary degraders of wood polyphenolic lignin. The ligninolytic enzymes produced by mushroom, including manganese peroxidase, lignin peroxidase, and laccase, mediate the oxidative degradation of lignin. The catalytic power of these enzymes in the degradation of aromatic ring compounds has been sought for the degradation of various organic compounds. In this project, we have screened 60 wild mushroom strains for their degradation activity against two representative PAHs, naphthalene and anthracene, and five aromatic dyes, including alizarin red S, crystal violet, malachite green, methylene blue, rose bengal. The degradation of PAHs was measured by GC while the decolorization of dyes was measured by both UV spectrophotometer and HPLC. As results, 9 wild mushroom strains showed high activity in degradation of PAHs and textile dyes. We also describe the secretive enzyme activities, the transcription levels, and cloning of target genes. In conjunction with this, activities of degradative enzymes, including laccase, lignin peroxidase, and Mn peroxidase, were measured in the liquid medium in the presence of PAHs and dyes. Our results showed that the laccase activity was directed correlated with the degradation, indicating that the main enzyme acts on PAHs and dyes is the laccase. The laccase activity was further simulated by the addition of $Cu^{2+}$ ion. Detailed studies of the enzyme system should be sought for future applications.

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Purification and Characterization of an Intracellular NADH: Quinone Reductase from Trametes versicolor

  • Lee, Sang-Soo;Moon, Dong-Soo;Choi, Hyoung-T.;Song, Hong-Gyu
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.333-338
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    • 2007
  • Intracellular NADH:quinone reductase involved in degradation of aromatic compounds including lignin was purified and characterized from white rot fungus Trametes versicolor. The activity of quinone reductase was maximal after 3 days of incubation in fungal culture, and the enzyme was purified to homogeneity using ion-exchange, hydrophobic interaction, and gel filtration chromatographies. The purified enzyme has a molecular mass of 41kDa as determined by SDS-PAGE, and exhibits a broad temperature optimum between $20-40^{\circ}C$, with a pH optimum of 6.0. The enzyme preferred FAD as a cofactor and NADH rather than NADPH as an electron donor. Among quinone compounds tested as substrate, menadione showed the highest enzyme activity followed by 1,4-benzoquinone. The enzyme activity was inhibited by $CuSO_4,\;HgCl_2,\;MgSO_4,\;MnSO_4,\;AgNO_3$, dicumarol, KCN, $NaN_3$, and EDTA. Its $K_m\;and\;V_{max}$ with NADH as an electron donor were $23{\mu}M\;and\;101mM/mg$ per min, respectively, and showed a high substrate affinity. Purified quinone reductase could reduce 1,4-benzoquinone to hydroquinone, and induction of this enzyme was higher by 1,4-benzoquinone than those of other quinone compounds.

Stabilization of High Nickel Cathode Materials with Core-Shell Structure via Co-precipitation Method (공침법을 통하여 합성된 코어-쉘 구조를 가지는 하이 니켈 양극 소재 안정화)

  • Kim, Minjeong;Hong, Soonhyun;Jeon, Heongkwon;Koo, Jahun;Lee, Heesang;Choi, Gyuseok;Kim, Chunjoong
    • Korean Journal of Materials Research
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    • v.32 no.4
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    • pp.216-222
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    • 2022
  • The capacity of high nickel Li(NixCoyMn1-x-y)O2 (NCM, x ≥ 0.8) cathodes is known to rapidly decline, a serious problem that needs to be solved in a timely manner. It was reported that cathode materials with the {010} plane exposed toward the outside, i.e., a radial structure, can provide facile Li+ diffusion paths and stress buffer during repeated cycles. In addition, cathodes with a core-shell composition gradient are of great interest. For example, a stable surface structure can be achieved using relatively low nickel content on the surface. In this study, precursors of the high-nickel NCM were synthesized by coprecipitation in ambient atmosphere. Then, a transition metal solution for coprecipitation was replaced with a low nickel content and the coprecipitation reaction proceeded for the desired time. The electrochemical analysis of the core-shell cathode showed a capacity retention of 94 % after 100 cycles, compared to the initial discharge capacity of 184.74 mA h/g. The rate capability test also confirmed that the core-shell cathode had enhanced kinetics during charging and discharging at 1 A/g.

Synthesis and characterization of sulfonated poly(arylene biphenylsulfone ether) copolymers containing hydroquinone moiety for polymer electrolyte membrane (고분자 전해질 멤브레인용 하이드로퀴논 부분이 포함된 설폰화된 폴리(아릴렌 비페닐설폰 에테르) 공중합체의 합성과 특성평가)

  • Yoo, Dong-Jin
    • Journal of Energy Engineering
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    • v.19 no.2
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    • pp.121-127
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    • 2010
  • In present work, sulfonated poly(arylene biphenyklsulfone ether) copolymers containing hydroquinone moiety were successfully synthesized using 4,4'-bis[(4-chlorophenyl)sulfonyl]-1,1'-biphenyl(BCPSBP), hydroquinone sulfonic acid potassium salt(sHQ), 4,4'-sulfonyldiphenol and evaluated their characteristics. Three kinds of polymer electrolyte membranes, PBPSEH-HQ00, PBPSEH-HQ10 and PBPSEH-HQ30 were prepared by using mole fraction of sulfonated hydroquinone(sHQ). The structure of the fabricated polymers was analyzed using NMR, IR and GPC. The Mw(weight-average molecular weights) of the polymers were in the range of 62,000-213,000 g $mol^{-1}$, and the molecular weight distribution (Mw/Mn) varied from 1.66-4.04. The thermal analysis of the copolymers was carried out by TGA and DSC. The temperature of Td5% and Td10% was decreased with the mole fraction of sHQ but Tg was increased with the mole fraction. The water uptake, IEC and ion conductivity were increased with increasing the ionic cluster of the polymers. The proton conductivity equal to 9.4 mS $cm^{-1}$ was measured for the PBPSEH-HQ30 membrane at $90^{\circ}C$ and 100% relative humidity. From the observed results it is clear that the prepared hydrocarbon membrane can be considered as suitable polymer electrolyte membrane for the application of PEMFC.

Environmental effects from Natural Waters Contaminated with Acid Mine Drainage in the Abandoned Backun Mine Area (백운 폐광산의 방치된 폐석으로 인한 주변 수계의 환경적 영향)

  • 전서령;정재일;김대현
    • Economic and Environmental Geology
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    • v.35 no.4
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    • pp.325-337
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    • 2002
  • We examined the contamination of stream water and stream sediments by heavy metal elements with respect to distance from the abandoned Backun Au-Ag-Cu mine. High contents of heavy metals (Pb, Zn, Cu, Cd, Mn, and Fe) and aluminum in the waters connected with mining and associated deposits (dumps, tailings) reduce water quality. In the mining area, Ca and SO$_4$ are predominant cation and anion. The mining water is Ca-SO$_4$ type and is enriched in heavy metals resulted from the weathering of sulfide minerals. This mine drainage water is weakly acid or neutral (pH; 6.5-7.1) because of neutralizing effect by other alkali and alkaline earth elements. The effluent from the mine adit is also weakly acid or neutral, and contains elevated concentrations of most elements due to reactions with ore and gangue minerals in the deposit. The concentration of ions in the Backun mining water is high in the mine adit drainage water and steeply decreased award to down stream. Buffering process can be reasonably considered as a partial natural control of pollution, since the ion concentration becomes lower and the pH value becomes neutralized. In order to evaluate mobility and bioavailability of metals, sequential extraction was used for stream sediments into five operationally defined groups: exchangeable, bound to carbonates, bound to FeMn oxide, bound to organic matter, and residual. The residual fraction was the most abundant pool for Cu(2l-92%), Zn(28-89%) and Pb(23-94%). Almost sediments are low concentrated with Cd(2.7-52.8 mg/kg) than any other elements. But Cd dominate with non stable fraction (68-97%). Upper stream sediments are contaminated with Pb, and down area sediments are enriched with Zn. It is indicate high mobility of Zn and Cd.