• Title/Summary/Keyword: ammonium ion

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Purification and Characterization of Lipase from Acinetobacter sp. B2 Isolated from Oil­contaminated Soil (유류오염지역에서 분리한 Acinetobacter sp. B2로부터의 Lipase 정제 및 특성)

  • Son Seung Hwa;Park Kyeong Ryang
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.320-327
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    • 2004
  • Three hundreds thirty two bacterial colonies which were able to degrade crude oil were isolated from soil sam­ples that were contaminated with oil in Daejeon area. Among them, one bacterial strain was selected for this study based on its higher oil degrading ability, and this selected bacterial strain was identified as Acinetobactor sp. B2 through physiological-biochemical tests and analysis of its 16S rRNA sequence. Acinetobactor sp. B2 was able to utilize various carbohydrates but did not utilize trehalose and mannitol as a sole carbon source. Acinetobactor sp. B2 showed a weak resistance to antibiotics such as kanamycin, streptomycin, tetracycline and spectinomycin, but showed a high resistance up to mg/ml unit to heavy metals such as Ba, Li, Mn, AI, Cr and Pb. The optimal growth temperature of Acinetobactor sp. B2 was $30^{\circ}C.$ The lipase produced by Acinetobactor sp. B2 was purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Its molecular mass was about 60 kDa and condition for the optimal activity was observed at $40^{\circ}C$ and pH 10, respectively. The activation energy of lipase for the hydrolysis of p­nitrophenyl palmitate was 2.7 kcal/mol in the temperature range of 4 to $37^{\circ}C,$ and the enzyme was unstable at the temperature higher than $60^{\circ}C.$ The Michaelis constant $(K_m)\;and\;V_{max}$ for p-nitrophenyl palmitate were 21.8 uM and $270.3\;{\mu}M\;min^{-1}mg^{-1},$ respectively. This enzyme was strongly inhibited by 10 mM $Cd^{2+},\;Co^{2+},\;Fe^{2+},\;Hg^{2+},$ EDTA and 2-Mercaptoethalol.

Characterization of a Fibrinolytic Enzyme from Pickled Anchovy (멸치젓갈유래의 혈전용해호소에 대한 특성)

  • Yang Woong-Suk;Lim Hak-Seob;Chung Kyung Tae;Kim Young-Hee;Huh Man Kyu;Choi Byung Tae;Choi Yung Hyun;Jeong Yong Kee
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.434-438
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    • 2005
  • In the previous study, we isolated a myulchikinase (MK), which has fibrinolytic activity and cytotoxicity to the tumor cell line, from myl- chi-jeot-gal. In this study, the effect of NaCl concentration, metallic ions, pH, temperature, and plasminogen on the activity of MK was analysed. The MK activity was maintained at least $80\%$ activity up to $30\%$ NaCl, which indicates that the enzyme may be halotolerant. The optimal pH and temperature were 8 and $40^{\circ}C$, respectively. The fibrinolytic activity of MK was completely inhibited with 0.5 mM $Hg^{2+}$ and inhibited to $50^{\circ}C$ with 1 mM $Cu^{2+}\;and\;Zn^{2+}$. The MK showed strong activity in plasminogen- rich fibrin plate but not in plasminogen-free fibrin plate. The result indicates that the MK may be a plasminogen activator type fibrinolytic enzyme.

Thermostable $\alpha$-Amylase Production by Thermophilic Bacillus sp. TR-25 lsolated from Extreme Enviroment (극한환경에서 분리한 고온성 Bacillus sp. TR-25에 위한 내열성 $\alpha$-amylase의 생산)

  • 노석범;손홍주;이종근
    • Journal of Life Science
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    • v.7 no.1
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    • pp.30-38
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    • 1997
  • For screening thermostable $\alpha$-amylase from thermophiles, various samples from extreme environments such as hot spring and sewage near them, and compoat, wereexamined microbial growth in enrichment culture medium at 55$\circ$C on the assumption that enzymes from thermophiles are inevitable thermostable. One strain showing higher $\alpha$-amylase activity was pure cultured and designated as Bacillus sp. TR-25 from the results of morphological, cultural and physiological characteristics. The most important carbon sourses for the enzyme production were soluble starch, dextrin, potato starch and corn starch. Glucose and fructose had a catabolite repression on the enzyme production. The good nitrogen sources for the enzyme production were yeat extract, nutrient broth, tryptone, corn steep liquor and ammonium sulfate. The enzyme production was accelerated by addition of CaCl$_{2}$. $\cdot $ H$_{2}$O. The optimal medium composition for the enzyme production was soluble starch 2.0%, yeast extract 0.55, CaCl$_{2}$ $\cdot $ 2H$_{2}$O 0.015, Tween 80 0.001%, pH8.0, respectively. In jar fermenter culture, this strain shows a rapid growth and required cheaper carbon and nitrogen source. These properties are very useful to fermentation industry. The $\alpha$-amylase of this strain demonstrated a maximum activity at 80$\circ$C, pH 5.0, respectively. And calcium ion did not improve thermostability of the enzyme. At 10$0^{\circ}C$, this enzyme has 235 of relative activity. Transformation was carried out by thermophilic Bacillus sp. TR-25 genomic DNA. As a result, the transformant has increased thermostable $\alpha$-amylase activity.

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Asparagine Biosynthesis in Soybean Sprouts (콩나물의 Asparagine 생합성(生合成)에 관(關)한 연구(硏究))

  • Byun, Si-Myung;Huh, Nam-Eung;Lee, Chun-Yung
    • Applied Biological Chemistry
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    • v.20 no.1
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    • pp.33-42
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    • 1977
  • Asparagine biosynthesis by soybean sprouts grown under the dark conditions has been demonstrated. The amount of free asparagine synthesized in ten day-old soybean sprouts increases to 22.7% on the dry weight base. The effects of nitrogen compounds such as $NH_4Cl,\;(NH_4)_2SO_4$ and urea on asparagine synthesis during the sprouting were examined and the results showed that urea was more effective than other two compounds. Glutamine-dependent asparagine synthetase was partially purified (8.6 folds) through ammonium sulfate fractionation, followed by Sephadex G-150 gel filtration. The enzyme was very labile and required protection by thiol groups or high level of glycerol. The mixture of ATP and $Mg^{++}$ ion also stabilized the enzyme activity. The enzyme utilized glutamine more effectively than ${NH_4}^+$ as an amide donor for the formation of asparagine. The enzyme required L-aspartate (Km=3.1 mM), L-glutamine, ATP and $Mg^{++}$. It showed pH optimum of 7.5 and catalyzed the formation of ${\beta}-aspartyl$ hydroxamate in the presence of L-aspartate, ATP, $Mg^{++}$ and $NH_2OH$ in the reaction mixture.

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Characterization of an Aniline-degrading Bacterium, Delftia sp. JK-2 Isolated from Activated Sludge of Municipal Sewage Treatment Plant (도시폐수처리장의 활성슬러지에서 분리한 Aniline 분해세균 Delftia sp. JK-2의 특성연구)

  • 조윤석;강형일;장효원;오계현
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.79-83
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    • 2000
  • Activated sludge samples were collected from a municipal sewage treatment plant and used for enrichment of microbial consortia with aniline as the sole carbon and nitrogen source. Threc aniline-degrading bacteria were obtained lrom microbial consortia and an isolate which has excellent aniline degradability was selected for this study. The isolate was Gram-negative, and identified and designated as Delfha sp. JK-2 on the basis of various physiological and biochemical tests. 10 mM aniline was completely degraded within 24 hours after inoculation of the culture. Ammonium ion was liberated in the medium transiently during the incubation and disappeared when aniline was completely degraded. Addition of glucose as a supplementary source to aniline minimal media showed significant decrease in aniline degradat~on rate for the strain Effective degradation of aniline was achieved by the addition of 0.5% nitrate as a nitrogen source, and resulted in approximately 80% higher aniline degradation compared to the absence of nitrate. Phylogenetic analysis based on 16s [DNA sequence revealed that the strain was closely related to De@ia acidovorans, with 96% overall similarity. The 16s [DNA sequence of JK-2 was also found to be closely related to those of six other clonal types, including Acidovoru, Aquaspirillum. Xylophilus, Variovorm, and Rhodofernr.

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Purification and Properties of a Cysteinylglycinase from Proteus mirabilis (Proteus mirabilis가 생산하는 Cysteinylglycinase의 정제 및 성질)

  • Choi, Shin-Yang;Yu, Ju-Hyun;Hidehiko Kumagai;Tatsrokuro Tochikura
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.92-97
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    • 1988
  • Cysteinylglycinase was partially purified from Proteus mirabilis by consecutive procedure. The specific activity was increased about 16-fold to that of cell-free extract. The enzyme was found rather unstable on ammonium sulfate precipitation ann the precipitated enzyme protein became partially insoluble during dialysis. The precipitated enzyme was found to be solubilized by treatment of 4% Triton X-100 effectiviely, The optimum temperature and pH of the enzyme activity were 35$^{\circ}C$ and 7.3, respectively. After heat treatment of the enzyme at 5$0^{\circ}C$ for 30 min, it lost the activity to 70%. The enzyme was stable at pH 7.0-8.0. The molecular weight of the cysteinylglycinase was found to be about 190,000 by Sephadex G-150 gel filtration. The enzyme was activated by the addition of Mn$^{2+}$ and $Mg^{2+}$ ions. The maximal activation was obtained in preincubation with $Mg^{2+}$ ion for 30 min. The enzyme catalyzed the hydrolysis of various dipeptides and tripeptides. The Km and Vmax values for cysteinylglycine were 1.60 mM and 0.24 m unit/ mg, respectively.

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Partial Purification and Some Properties of Amylases from Germinating Corn(Zea mays L.) (발아 옥수수 amylases의 정제 및 특성)

  • Lee, Tae-Ho;Jung, Tae-Yung;Park, Mi-yeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.6
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    • pp.625-635
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    • 1990
  • The purpose of this study was focused on investigation of biochemical properties of amylases in germinating corn(Zea mays L.) the amylase(I), (II) and (III) from germinating corn seeds were partially purified by ammonium sulfate precipitation, DEAE-Sephadex A-50 ion exchange column chromatography and Sephadex G-100 gel filtration. The last step was effective for separation of the corn amylases to a homogeneous slate. the purified amylase(I) was identified as a kind of $\alpha$-amylase from the fact that 5% starch solution was hydrolysed into mainly maltose and maltotetrose by it, and amylase(II) and amylase(III) were enzymes producing maltotetrose as main product. The molecular weight and specific activity of the amylase(I), (II) and (III) were determined to be 54,000 and 70.47 unit/mg, 39,000 and 62.98 unit/mg, and 51,000 and 80.39 unit/mg, respectively. It showed a tendency to increase the amylases activities in presence of Ba, Ca, Co and Fe groups, but inhibits in that of Ag, Sn, Hg and Zn groups, and amylase(I), (II) and (III) remained stable at pH 5-6 and 2$0^{\circ}C$ for 40 days in containing of 1 mM CaCl$_2$. The optimum pH and optimum temperatures were pH 6, pH 5 and pH 6 and 35$^{\circ}C$, 55$^{\circ}C$ and 55$^{\circ}C$, respectively. These results suggest that the amylase(I), (II) and (III) were different amylases.

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Construction and Optimization of Selective Membrane Electrodes for Determination of Doxepin Hydrochloride in Pharmaceutical Preparations and Biological Fluids (약의 조제와 생물학적 유체에서 독스핀 하이드로클로라이드의 확인을 위한 선택적 막 전극의 구성과 최적화)

  • El-Tohamy, Maha;Razeq, Sawsan;El-Maamly, Magda;Shalaby, Abdalla
    • Journal of the Korean Chemical Society
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    • v.54 no.2
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    • pp.198-207
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    • 2010
  • The construction and performance characteristics of doxepin hydrochloride selective electrodes were developed. Three types of electrodes: plastic membrane I, coated wire II, and coated graphite rod III were constructed based on the incorporation of doxepin hydrochloride with ammonium reineckate. The influence of membrane composition, kind of plasticizer, pH of the test solution, soaking time, and foreign ions on the electrodes was investigated. The electrodes showed a Nernstain response with a mean slope of 57.41 ${\pm}$ 0.5, 56.22 ${\pm}$ 0.2 and 52.88 ${\pm}$ 0.7 mV at $25^{\circ}C$ for electrode I, II and III respectively, over Doxepin hydrochloride concentration range from $1{\times}10^{-2}-1{\times}10^{-6}M$, $5{\tims}10^{-2}-1{\times}10^{-6}M$ and $1{\times}10^{-3}-5{\times}10^{-6}M$, and with a detection limit $5.0{\times}10^{-7}M$, $6.3{\times}10^{-7}M$ and $2.5{\times}10^{-6}M$ for electrode I, II and III respectively. The constructed electrodes gave average selective precise and usable within the pH range 3 - 7. Interferences from common cations, alkaloids, sugars, amino acids and drug excipients were reported. The results obtained by the proposed electrodes were also applied successfully to the determination of the drug in pharmaceutical preparations and biological fluids.

Purification and Characterization of Chinese Cabbage Pectinesterase (배추 펙틴에스테라제의 정제 및 특성)

  • Ko, Young-Hwan;Park, Kwan-Hwa
    • Korean Journal of Food Science and Technology
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    • v.16 no.2
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    • pp.235-241
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    • 1984
  • Two fractions of pectinesterase from Chinese cabbage were isolated by ammonium sulfate fractionation, ion exchange chromatography on DEAE-cellulose and Sephadex G-150 gel filtration. The fraction F-A and F-B were purified approximately 340- and 10-fold. The similar salt effects and pH optima (pH 7.5-8.0) were obtained for the two pectinesterase fractions. The maximum activity of both two. fractions were obtained at 20-50mM of divalent rations and at 250mM of monovalent rations. The apparent Michaelis constant of the F-A was 0.01% for citrus pectin. The temperature optima for F-A and F-B were $48^{\circ}$ and $55^{\circ}C$, respectively and both fractions were stable in the region of pH 5.0-8.0 at room temperature. The thermal inactivation of the two fractions followed the first order reaction kinetics. From D and Z-values obtained the thermal resistance of the two fractions were characterized.

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Chemical Composition of Fine Particulate Matter in the Downtown Area of Jeju City (제주시 도심지역 미세먼지의 화학적 조성 특성)

  • Hu, Chul-Goo;Lee, Ki-Ho
    • Journal of Environmental Science International
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    • v.27 no.7
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    • pp.597-610
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    • 2018
  • This study observed particulate matter ($PM_{2.5}$ and $PM_{10}$) in the downtown area of Jeju City, South Korea, to understand the chemical composition of particulates based on an analysis of the water-soluble ionic species contained in the particles. The mass fraction of the ionic species in the sampled $PM_{10}$ and $PM_{2.5}$ was 44.3% and 42.2%, respectively. In contrast, in Daegu City and Suwon City, the mass fraction of the ionic species in $PM_{2.5}$ was higher than that in $PM_{10}$. The chloride depletion percentage of $PM_{10}$ and $PM_{2.5}$ in Jeju City was higher than 61% and 66%, respectively. The contribution of sea-salt to the mass of $PM_{10}$ (5.9%) and $PM_{2.5}$ (2.6%) in Jeju City was similar to that in several coastal regions of South Korea. The mass ratio of $Cl^-$ to $Na^+$ in the downtown area of Jeju City was comparable to that in some coastal regions, such as the Gosan Area of Jeju Island, Deokjeok Island, and Taean City. The mass fraction of sea-salt in $PM_{10}$ and $PM_{2.5}$ was very low, and the concentration of sodium and chloride ions in $PM_{10}$ was not correlated with those in $PM_{2.5}$ ($R^2$ < 0.2), suggesting that the effects of sea-salt on the formation of particulate matter in Jeju City might be insignificant. The relationship between $NH_4{^+}$ and several anions such as $SO_4{^{2-}}$, $NO_3{^-}$, and $Cl^-$, as well as the relationship between the measurement and calculation of ammonium ion concentration, suggested that sea-salts may not react with $H_2SO_4$, and $(NH_4)_2SO_4$ may be a major secondary inorganic aerosol component of $PM_{2.5}$ and $PM_{10}$ in Jeju City.