• Title/Summary/Keyword: Zn$^{+2}$/Fe$^{+2}$ 이온

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Purification and Characterization of Catechol 2,3-Dioxygenase from Recombinant Strain E. coli CNU312. (재조합균주 E. coli CNU312가 생산하는 Catechol 2,3-Dioxygenase의 정제 및 특성)

  • 임재윤;최경호;최병돈
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.26-32
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    • 2000
  • Catechol 2,3-dioxygenase was purified from recombinant strain E. coli CNU312 carrying the tomB gene which was cloned from toluene-degrading Burkholderia cepacia G4. The purification of this enzyme was performed by acetone precipitation, Sephadex G-75 chromatography, electrophoresis and electro-elution. The molecular weight of native enzyme was about 140.4 kDa and its subunit was estimated to be 35 kDa by SDS-PAGE. It means that this enzyme consists of four identical subunits. This enzyme was specifically active to catechol, and$K_(m)$ value and $V_(max)$value of this enzyme were 372.6 $\mu$M and 39.27 U/mg. This enzyme was weakly active to 3-methylcatechol, 4-methylcatechol, and 4-chlorocatechol, but rarely active to 2,3-DHBP. The optimal pH and temperature of the enzyme were pH 8.0 and $40^{\circ}C$. The enzyme was inhibited by $Co^(2+)$, $Mn^(2+)$, $Zn^(2+)$, $Fe^(2+)$, $Fe^(3+)$, and $Cu^(2+)$ ions, and was inactivated by adding the reagents such as N-bromosuccinimide, and $\rho$-diazobenzene sulfonic acid. The activity of catechol 2,3-dioxygenase was not stabilized by 10% concentration of organic solvents such as acetone, ethanol, isopropyl alcohol, ethyl acetate, and acetic acid, and by reducing agents such as 2-mercaptoethanol, dithiothreitol, and ascorbic acid. The enzyme was inactivated by the oxidizing agent $H_(2)$$O_(2)$, and by chelators such as EDTA, and ο-phenanthroline.

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Characteristics of the Extracellular Proteolytic Enzyme Produced by Candida lipolytica FM5 Isolated from Mackerel (Scomber japonicus) (Candida lipolytica FM5가 생산하는 균체외 단백분해효소의 특성)

  • PARK Mi-Yeon;OH Eun-Gyong;CHANG Dong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.5
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    • pp.631-636
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    • 1998
  • Candida lipolytica(C. lipolytica) FM5 was selected as one of the strong saprophytic yeasts isolated from mackerel (Scomber japonicus). The selected strain could produce extracellular proteolytic enzyme. The effective medium for production of proteolytic enzyme by C. lipolytica FM5 was TPPY broth containing Bacto-tryptone $0.5\%$, proteose peptone $0.5\%$, yeast extract $0.25\%$, NaCl $0.5\%$ and $CaCl_2\;0.2\%$. The pattern of proteolytic enzyme production by C. lipolytica FM5 was the almost same as that of growth curve of the strain. Namely, the enzyme production was begun from the early stage of exponential phase and it was reached the highest at the begining of the stationary phase of the yeast growth. The optimum toeperature of the produced proteolytic enzyme was $35^{\circ}C$ and its activity was not significantly changed by pH between 6.5$\~$9.0 and also it was not significantly affected by several kinds of cations such as $Ca^{2+},\;Cu^{2+},\;Fe^{2+}$ and $Mg^{2+}$ but it was affected negatively by some cations such as $Zn^{2+},\;Mn^{2+}$ and $K^+$.

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Purification and Characterization of the Extracellular Alginase Produced by Bacillus licheniformis AL-577 (알긴산 분해균 Bacillus licheniformis AL-577가 생산하는 균체외 효소의 정제 및 특성)

  • Uo, Meung-Hee;Joo, Dong-Sik;Cho, Soon-Yeong;Min, Tae-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.2
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    • pp.231-237
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    • 2006
  • The extracellular enzyme alginase produced by Bacillus licheniformis AL-577 was purified by ion chromatography on CM-Cellulose column, DEAE-Sepharose column, and followed by gel filtration on Sephadx G-100 column. The optimum pH and temperature for the activity of the purified enzyme were 6.0 and $35^{\circ}C$, respectively. The enzyme was stable at the pH range of $6.0\~9.0$ and at $20^{\circ}C$. The molecular weight of the enzyme was estimated to be about 25,500 daltons by SDS-polyacrylamide gel electrophoresis. NaCl was required for high activity of the enzyme. The enzyme was inhibited by $Ba^{2+},\;Co^{2+},\;Cu^{2+},\;Fe^{2+},\;Mg^{2+},\;Zn^{2+},\;NH_4^+$, EDTA, L-cysteine, and 2-mercaptoethanol, while stimulated by DTT, O-phenanthroline, $K^+$ and $Li^+$. This enzyme was proposed to be an alginase specifically degrading alginic acid.

Purification and Characterization of Polyphenol Oxidase from Flammulina velutipes (팽나무버섯 polyphenol oxidase의 정제 및 특성)

  • Pyo, Han-Jong;Son, Dae-Yeul;Lee, Chan
    • Korean Journal of Food Science and Technology
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    • v.34 no.4
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    • pp.552-558
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    • 2002
  • Polyphenol oxidase from Flammulina velutipes was purified and characterized. Purification of polyphenol oxidase was achieved by ammonium sulfate precipitation, Superdex G-200 gel filtration chromatography, Phenyl superose affinity chromatography, Mono-Q anion exchange chromatography and Superdex S-200 gel filtration chromatography on FPLC. After these purification steps specific activity of purified polyphenol oxidase increased to 199.1 units/mg. Polyphenol oxidase from F. velutipes was composed of a single polypeptide with molecular weight of about 40 kDa. Optimum pH and temperature for the enzyme reaction were found to be 6.0 and $25^{\circ}C$, respectively. The activity of the enzyme gradually decreased at acidic pH between 3 and 5, and the enzyme lost its activity at alkaline pH between 8 and 10. This enzyme exhibited high substrate specificity to o-diphenols. Km-values for L-DOPA and caffeic acid were found to be 3.97 mM and 1.78 mM, respectively. 2-mercaptoethanol, L-ascorbic acid, sodium bisulfite, EDTA and $Mg^{2+}$ inhibited the activity of pholyphenol oxidase and $Cu^{2+}$, $Fe^{2+}$, $Zn^{2+}$ and $Ni^{2+}$ increased enzyme activity. The activity of enzyme was well maintained at $-70^{\circ}C$ for over 4 months, and at $-20^{\circ}C$ for 1 months.

Mineralogy and Chemical Properties according to Particle Size Separation of Hwangto (Reddish Residual Soil) used in Feeding of Cattle (한우 사육에 이웅한 황토(풍화토)의 입도분리에 따른 광물성분 및 화학적 특성)

  • 황진연;박현진;양경희;이효민
    • Journal of the Mineralogical Society of Korea
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    • v.15 no.1
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    • pp.33-43
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    • 2002
  • Mineral composition and chemical properties of Hwangto (reddish residual soil) that used in feeding of cattles at Iksan, Jeollabuk-do, Korea were examined according to particle size separation such as gravel, sand, silt, coarse clay and fine clay. Mineral composition analyses reveal that gravel and sand are mainly composed of quartz and feldspars and that kaolin mineral and illite are dominant in clay and silt. Iron oxides are mainly included in fine clay. According to chemical analyses of major elements, Al, Fe and $H_2O$ contents are increased with decreasing of particle size. This trend well agrees with increase of clay minerals in smaller particles, Chemical analyses of trace elements indicate that contents of Zn, Rb, Sr, Ba, Pb significantly differ with particle sizes. Ba and Sr are included in feldspars since these elements are abundant in sand containing abundant feldspars. Pb and Sm are abundant in sample before particle size separation, but the contents are significantly decreased after separation. Therefore, most of these elements appear to be existed as removable phase. Nb, La, Th, Ce are more abundant in silt. The contents of all the other trace elements tend to be increased in smaller particles containing more clay minerals. The contents of changeable cations and teachable elements in acid and alkali solutions are high in clay samples. All the above results indicate that using the portion of smaller particle of Hwangto for livestock feed rather than bulk Hwangto can improve cation exchangeable capacity, ion leaching capacity and sorption properties.

The Evaluation of Trace Minerals Levels of Pasture Soils and Forages in Jeju (제주지역 목장 토양 및 조사료 자원의 미량광물질 함량 평가)

  • Lee, Chong-Eon;Park, Myung-Hee;Park, Nam-Keon;Park, Hyung-Soo;Oh, Woon-Yong
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.27 no.1
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    • pp.29-36
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    • 2007
  • A study was conducted to determine the trace minerals (Cu, Fe, Cd, Zn, Mo, Se, Mn, Cr, Co, Ni) levels in pasture soils and forages collected in Jeju area. Chemical characteristics and total or soluble trace minerals levels in 187 pasture soil samples (76 very dark brown and 111 black soils) were measured. Total trace minerals contents in hay samples of 60 Italian ryegrass, alfalfa and mixture grasses each were assessed. The pasture soils of Jeju were characterized to have low pH (5.1) and to contain low levels of available $P_2O_5$ (20.5 mg/kg) and exchangeable Ca, Mg, K, Na (2.6, 0.9, 0.5, 0.2 cmol+/kg, respectively) when comparing to upland soils of Jeju or Korean mainland. All trace elements in total or soluble analysis of pasture soils were detected, and there was a big difference between total and soluble levels. The pasture soils tended to have the higher total Fe, Mn, Ni and Zn contents. Cr in all forages was not detected, but other trace minerals levels showed normal range. The hay samples of Italian ryegrass and mixture grasses produced in Jeju tended to contain higher Mn (105 vs 23 mg/kg) and lower Mo (2.7 vs 4.9 mg/kg) than those of alfalfa hay imported from USA. Results show that trace minerals of pasture soils and forages in Jeju seem to be not deficient, indicating that supplementation of some trace minerals are not always necessary in diets for grazing animals and should be done after careful evaluation of diets with regard to concentrations and biological availability of essential elements.

Relationship between Physicochemical Properties, Heavy Metal Contents and Magnetic Susceptibility of Soils (토양의 물리화학적 특성, 중금속 함량, 대자율 간의 상호관계 연구)

  • Chon, Chul-Min;Park, Jeong-Sik;Kim, Jae-Gon;Lee, Youn-Soo
    • Journal of the Mineralogical Society of Korea
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    • v.23 no.4
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    • pp.281-295
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    • 2010
  • This paper deals with magnetic susceptibility, mineralogy, soil properties (pH, EC, CEC, loss on ignition), iron and manganese oxides, the content and partitioning of heavy metals (As, Cd, Cr, Cu, Ni, Pb, Zn), and their mutual relationship in the soil samples of an unpolluted, abandoned mine area, and industrial complex area. The various minerals derived from weathered bedrock were identified by X-ray diffraction in the unpolluted soil samples, except for the magnetic minerals. XRD analysis also revealed the existence of hematite and magnetite related to mine tailings and waste rocks in the abandoned mine area samples. The industrial complex area samples had carbonate minerals, such as calcite and dolomite, that might be due to anthropogenic deposition. The sum of the reducible, oxidizable, and residual fractions was over 80% for the abandoned mine area samples and over 50% for the industrial complex area samples using the sequential extraction method. The industrial complex area samples had a relatively high carbonate fraction that was associated with carbonate minerals. The content of aqua regia-extractable Fe, Mn, As, and Zn had a high positive correlation with the content of the dithionite-citrate-bicarbonate (DCB)-extractable method related to Fe/Mn oxide phases. The 54% and 58% of aqua regia-extractable Fe and As content, respectively, acted together with the concentrations of the DCB-extractable phases. Magnetic susceptibility values of total samples ranged from 0.005 to $2.131{\times}10^{-6}m^3kg^{-1}$. The samples including iron oxide minerals, such as hematite and magnetite, had a high magnetic susceptibility. The magnetic susceptibility showed a significant correlation with the heavy metals, Cd (r=0.544, p<0.05), Cr (r=0.714, p<0.01), Ni (r=0.645, p<0.05), Pb (r=0.703, p<0.01), and Zn (r=0.496, p<0.01), as well as Fe (r=0.608, p<0.01) and Mn (r=0.615, p<0.01). The aqua regia-extractable Fe and Mn content had a significant positive correlation with Cd, Cr, Cu, Ni, and Zn. However, the DCB-extractable Fe and Mn content had a significant positive correlation with As and Ni, indicating that the heavy metals were associated with Fe and Mn oxide minerals.

A study on the preparation of high purity nickel carbonate powders in solvent extraction processing solution from waste iron-nickel alloy etchant (철-니켈 합금 에칭구액 용매추출 공정 용액으로부터 고순도 탄산니켈 제조에 관한 연구)

  • Chae, Byung-man;Hwang, Sung-ok;Lee, Seok-Hwan;Kim, Deuk-Hyeon;Lee, Sang-Woo;Kim, Dae-Weon;Choi, Hee-Lack
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.27 no.6
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    • pp.303-308
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    • 2017
  • The $FeCl_3$ waste solution used to etch various metals contains valuable metal such as nickel. In this study, we recovered as high purity nickel carbonate crystalline powders from nickel-containing etching waste solution after regeneration of iron chloride. Firstly we eliminated about of the iron impurities under the condition of pH 4 using 5 % NaOH aqueous solution and then removed the remaining impurities such as Ca, Mn and Zn etc. by using solvent extractant D2EHPA (Di-(2-ethylhexyl) phosphoric acid). Thereafter, nickel carbonate powder having a purity of 99.9 % or more was obtained through reaction with sodium carbonate in a nickel chloride solution.

Wet Oxidation of Phenol with Homogeneous Catalysts (균일촉매를 이용한 페놀의 습식산화)

  • Suh, Il-Soon;Ryu, Sung Hun;Yoon, Wang-Lai
    • Korean Chemical Engineering Research
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    • v.47 no.3
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    • pp.292-302
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    • 2009
  • The wet oxidation of phenol has been investigated at temperatures from 150 to $250^{\circ}C$ and oxygen partial pressures from 25.8 to 75.0 bar with initial pH of 1.0 to 12.0 and initial phenol concentration of 10 g/l. Chemical Oxygen Demand COD has bee measured to estimate the oxidation rate. Reaction intermediates have been identified and their concentration profiles have been determined using liquid chromatography. The destruction rate of phenol have shown the first-order kinetics with respect to phenol and the changes in COD during wet oxidation have been described well with the lumped model. The impact of various homogeneous catalysts, such as $Cu^{2+}$, $Fe^{2+}$, $Zn^{2+}$, $Co^{2+}$ and $Ce^{3+}$ ions, on the destruction rate of phenol and COD has also been studied. The homogeneous catalyst of $CuSO_4$ has been found to be the most effective for the destruction of phenol and COD during wet oxidations. The destruction rate of formic acid formed during wet oxidations of phenol have increased as increasing temperature and $CuSO_4$ concentration. The final concentrations of acetic acid which has been formed during wet oxidations and difficult to oxidize have increased with reaction temperature and with decrease in the catalyst load.

Biochemical Characterization of a Protease with Fibrinolytic Activity from Maggots of Protaetia brevitarsis (Protaetia brevitarsis의 maggot로부터 fibrinolytic activity을 가진 protease의 생화학적 특성 연구)

  • Chang, Jeong-Hyun;Jo, Ji-Young;Kim, Yeong-Jin;Lee, Sun-Yi;Cho, Hyo-Jin;You, Sun-Nyoung;Kim, Kwang-Youn;Park, Byoung-Keun;Ahn, Soon-Cheol;Kwon, Heun-Young
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.606-612
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    • 2007
  • Fibrin clots remained in blood vessels can be one of the serious factor caused cardiovascular disease, such as ischemia, infarction and necrosis The development of an antithrombotic and thrombolysis solvent is necessary to prevent and treat these diseases. In this study, the fibirinolytic protease was prepared from the maggots of Protaetia brevitarsis using ammonium sulfate fractionation and desalting column. The optimum pH and temperature for the enzyme activity were pH 9.0 and $50^{\circ}C$, respectively. The enzyme activity was relatively stable at pH 7.0-9.0 and temperature below $60^{\circ}C$. The activity of the enzyme was strongly inhibited by phenylmethanesulfonyl fluoride. And the activity of the enzyme was inhibited by $Ca^{2+}\;and\;Zn^{2+}$, but it was not by $Mg^{2+}\;and\;Fe^{2+}$ ions. In these experimental results, we have speculated that the enzyme derived from maggots of Protaetia hrevitarsis is a serine protease with a strong fibrinolytic activity.