• Title/Summary/Keyword: Ca-EDTA

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Characterization of Calcium-Activated Bifunctional Peptidase of the Psychrotrophic Bacillus cereus

  • Kim Jong-Il;Lee Sun-Min;Jung Hyun-Joo
    • Journal of Microbiology
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    • v.43 no.3
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    • pp.237-243
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    • 2005
  • The protease purified from Bacillus cereus JH108 has the function of leucine specific endopeptidase. When measured by hydrolysis of synthetic substrate (N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide), the enzyme activity exhibited optimal activity at pH 9.0, $60^{\circ}C$. The endopeptidase activity was stimulated by $Ca^{++},\;Co^{++},\;Mn^{++},\;Mg^{++},\;and\;Ni^{++}$, and was inhibited by metal chelating agents such as EDTA, 1,10-phenanthroline, and EGTA. Addition of serine protease inhibitor, PMSF, resulted in the elimination of the activity. The endopeptidase activity was fully recovered from the inhibition of EDTA by the addition of 1 mM $Ca^{++}$, and was partially restored by $Co^{++}\;and\;Mn^{++}$, indicating that the enzyme was stabilized and activated by divalent cations and has a serine residue at the active site. Addition of $Ca^{++}$ increased the pH and heat stability of endopeptidase activity. These results show that endopeptidase requires calcium ions for activity and/or stability. A Lineweaver-Burk plot analysis indicated that the $K_m$ value of endopeptidase is 0.315 mM and $V_{max}$ is 0.222 ) is $0.222\;{\mu}mol$ of N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide per min. Bestatin was shown to act as a competitive inhibitor to the endopeptidase activity.

Characterization of Chinese Cabbage Phospholipase D by a Multistirring Batch System Bioreactor (다중 교반형 생물반응기에 의한 배추 Phospholipase D의 특성연구)

  • 박동훈;정의호이해익이상영
    • KSBB Journal
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    • v.5 no.2
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    • pp.119-124
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    • 1990
  • Phospholipase D catalyzes the phosphatidohydrolysis and transphosphatidylation of phospholipid in the biological systems. In this study we were partially purified phospholipase D from Chinese cabbage and the characterization of the enzyme was carried out in a multistirring batch system bioreactor. The enzyme showed optimum activity at pH ,5.6, highest activity at 37$^{\circ}C$ and Ca2+ is important for the enzyme activity. Optimum concentrations of Ca2+ for phosphatidohydrolysis was 20 mM and for transphosphatidylation was 40 mM, respectively. Some organic solvents such as diethylether, isopropylether and butylacetate were activated the enzyme activity. On the other hand, EDTA, Ba2+, Mn2+ and Zn2+ showed inhibitory effect on the enzyme activity. The base acceptors in transphosphatidylation by the Chinese cabbage phospholipase D were tested. Various poly-and monohydroxy alcohols were found to be active.

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Mercury Ion Removal Using a Packed-Bed Column with Granular Aminated Chitosan

  • JEON, CHOONC
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.497-501
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    • 2005
  • This study deals with the removal of mercury species using a packed-bed column with spherical aminated chitosan material. These adsorbents revealed a high adsorption capacity for mercury species. Experiments with feed solutions of 10 ppm Hg dissolved in distilled water showed an excellent removal with a sharp increase of the filter effluent concentration after a total throughput of 900 bed volumes of feed water. Up to $95\%$ desorption was reached by using 3 bed volumes of 0.01 N EDTA solution. EDTA could be recovered by means of sulfuric acid with about $75\%$ efficiency. Almost the same results were obtained in repeated sorption and desorption experiments at identical conditions. The experiments demonstrated that the sorbents possessed practically no sorption capacity for alkaline earth ions ($Ca^{2+}\;and\;Mg^{2+}$). Their influence on the sorption of mercury was negligible. In experiments with spiked tap water of the Karlsruhe Research Centre and a feed mercury concentration of 0.01 mg/l, the breakthrough of Hg was observed only after a total throughput of about 6,000 bed volumes of feed water.

Contents and Seasonal Variations of Arsenic in Paddy Soils and Rice Crops around the Abandoned Metal Mines (폐금속광산 주변 논토양 및 벼작물의 비소함량과 계절적 변화)

  • Kwon, Ji Cheol;Jung, Myung Chae;Kang, Man Hee
    • Economic and Environmental Geology
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    • v.46 no.4
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    • pp.329-338
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    • 2013
  • The objective of this study is to investigate the contents and seasonal variation of arsenic in soils and crop plant(rice) in paddy fields around the abandoned metal mines in Korea. The soils were extracted by various methods including aqua regia, 1 M $MgCl_2$, 0.01 M $CaCl_2$ and 0.05 M EDTA to evaluate the relationships between soils and crop plants(rice). According to correlation analysis, statistically significant correlation with the four methods(p<0.01) were found in soils extracted by various chemical solutions and arsenic contents in soils were decreased in the order of 1M $MgCl_2$ > 0.01M $CaCl_2$ > 0.05 M EDTA. Biological accumulation coefficients(BACs) of rice stalks were higher than those of rice grain, and the coefficients under reducing(August) environment were higher than those under oxidizing conditions(October). Assuming the rice consumption of 315 g/day by farm households in Korea, the amount of daily intake of arsenic were estimated to be 77.8 ${\mu}g/day$. The daily intake of arsenic from the rice estimates up to 65% of ADI(acceptable daily intake) that the FAO/WHO Joint Food Additive and Contaminants Committee has set to evaluate their safeties.

Studies on the Characterization of Protease Produced by Streptomyces alboniger (Streptomyces alboniger가 생산(生産)하는 Protease의 특성(特性)에 관(關)한 연구(硏究))

  • Kim, Kang-Shin;Han, Kang-Wan;Kim, Hyung-Rho
    • Applied Biological Chemistry
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    • v.27 no.3
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    • pp.174-179
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    • 1984
  • Some physico-chemical properties of proteolytic enzyme produced from Streptomyces alboniger Hesseltine were characterized. The optimal pH and temperature of the enzyme were around pH 9.0 and $40^{\circ}C$, respectively. The enzyme was stable between pH $6.0{\sim}10.8$ and the enzyme activity was not inactivated by heat treatment in lower temperature than $40^{\circ}C$. However the enzyme activity decreased by 70% of the initial activity for 10minutes at $70^{\circ}C$. The Km value was $7.1{\mu}M$ with Hammarsten casein and $333.3{\mu}M$ with cytochrom C. The activity of enzyme was inhibited by metal ions in the order of $Fe^{+++}>Hg^{++}>Cu^{++}>Pb^{++}>Zn^{++}$, whereas $Ca^{++}$ increased the enzyme activity. There was no effect of $Mg^{++}$ and $Co^{++}$ on the enzyme activity. The enzyme was inhibited by EDTA strongly. When $Ca^{++}$ was added to the EDTA-denaturated enzyme, the activity of enzyme was restored.

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The Effect of $Ca^{2+}$ and Its Specific Time in the Maturation of Mammalian Oocyte (포유류의 난자성숙과 성숙과정에서 $Ca^{2+}$의 영향과 그 작용시기)

  • Bae, In-Ha
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.3
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    • pp.285-296
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    • 1994
  • Follicular oocytes were released from the graafian follicles of ovaries from 3-4 weeks old mice. The spontaenous maturation of these follicular oocyes was inhibited by the treatment of dbcAMP and progesterone and these oocytes were cultured for 2-8hr in the Modified Hank's balanced salt solution(MHBS). Ethylenediaminetetraaceticacid(EDTA) and calmoudulin antagonist, trifluoperazine (TFP) were treated to the culture medium in order to investigate whether these chemical agents inhibit calcium uptake into the oocyte and oocyte maturation. $^{45}Ca^{2+}$, 10-${\mu}$Ci/ml was added to the culture medium during the culture period. $^{45}Ca^{2+}$uptake into the oocytes was examined whether and when various kind of oocyte maturation inhibiting agents inhibit or stimulate the influx of calcium into oocytes. Dibutyryl cAMP and progesterone decrease $^{45}Ca^{2+}$uptake into the oocytes and synergistic inhibiting effect of dbcAMP and progesterone was prominent at much lower dosages. Calcium uptake into oocytes seems to be higher during first 2 hour culture period rather than next 4hr culture. After 8hr culture, calcium uptake level of the oocytes which GVBD already took place gradually approached to the level of those which were maintained at GV by the treatment of dbcAMP and progesterone. However, $^{45}Ca^{2+}$uptake into the GV maintained oocytes did not change at all even after 8hr culture period. In addition, calcium chelating agent, EDTA inhibited calcium uptake into oocytes as well as nuclear maturation of oocytes. Lower dosage used in the present study did not inhibit calcium uptake as well as oocyte maturation.

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Application of Ferrate(VI) on the Decomplexation of Cu(II)-EDTA

  • Tiwari, Diwakar;Yang, Jae-Kyu;Chang, Yoon-Young;Lee, Seung-Mok
    • Environmental Engineering Research
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    • v.13 no.3
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    • pp.131-135
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    • 2008
  • In this study, Fe(VI) was employed as a multi-functional agent to treat the simulated industrial wastewater contaminated with Cu(II)-EDTA through oxidation of EDTA, decomplexation of Cu(II)-EDTA and subsequent removal of free copper through precipitation. The decomplexation of $10^{-4}\;M$ Cu(II)-EDTA species was performed as a function of pH at excess concentration of Fe(VI). It was noted that the acidic conditions favor the decomplexation of Cu(II)-EDTA as the decomplxation was almost 100% up to pH 6.5, while it was only 35% at pH 9.9. The enhanced degradation of Cu(II)-EDTA with decreasing the pH could be explained by the different speciation of Fe(VI). $HFeO_4^-$ and $H_2FeO_4$, which are relatively more reactive than the unprotonated species $FeO_4^{2-}$, are predominant species below neutral pH. It was noted that the decomplexation reaction is extremely fast and within 5 to10 min of contact, 100% of Cu(II)-EDTA was decomplexed at pH 4.0. However, at higher pH (i.e., pH 10.0) the decomplexation process was relatively slow and it was observed that even after 180 min of contact, maximum ca 37% of Cu(II)-EDTA was decomplexed. In order to discuss the kinetics of the decomplexation of Cu(II)-EDTA, the data was slightly fitted better for the second order rate reaction than the first order rate reaction in the excess of Fe(VI) concentration. On the other hand, the removal efficiency of free Cu(II) ions was also obtained at pH 4.0 and 10.0. It was probably removed through adsorption/coagulation with the reduced iron i.e., Fe(III). The removal of total Cu(II) was rapid at pH 4.0 whereas, it was slow at pH 10.0. Although the decomplexation was 100% at lower pH, the removal of free Cu(II) was relatively slow. This result may be explicable due to the reason that at lower pH values the adsorption/coagulation capacity of Fe(III) is greatly retarded. On the other hand, at higher pH values the decomplexation of Cu(II)-EDTA was partial, hence, slower Cu(II) removal was occurred.

Novel $Ca^{2+}$-ATPase Found in the Human Milk Membrane Fraction

  • Cho, Jin-Kook;Kanno, Choemon
    • 한국유가공학회:학술대회논문집
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    • 1997.05a
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    • pp.23-34
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    • 1997
  • Calcium-stimulated ATPase ($Ca^{2+}$-ATPase) which has optimal pH value at 7.0 was found in the membrane fraction of human milk, and its enzymatic properties were studied. The purified $Ca^{2+}$-ATPase required 0.45 mM Ca ion for maximal activity. Among the nucleosides, $Ca^{2+}$-ATPase showed a higher substrate specificity to ATP and UTP than to CTP and GTP. $Ca^{2+}$-ATPase had apparent Km value of 0.065, and V max of 7.63 mol ATP hydrolyzed/mg pro-tein per min, respectively. $Ca^{2+}$-ATPase was potently inhibited by lanthanide, vanadate, and p-chloromercuribenzoate, and inactivated by EDTA, and CDTA and EGTA, but were unaffected by N-ethylmaleimide, $NaN_3$, ouabain, or oligomycin, and was completely inactivated by heating at $60^{\circ}C$ for 10 min. This enzyme activity was concentrated in the membrane fraction of the cream and skim milk membrane, but not founded in bovine milk.

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Release of Cu from SDS micellar solution using complexing agents

  • 김호정;백기태;김보경;이율리아;양지원
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.307-310
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    • 2004
  • Micellar enhanced ultrafiltration (MEUF) is a surfactant-based separation process and it can remove heavy metal ions from aqueous stream effectively. However, it is necessary to recover and reuse surfactants for economic feasibility because surfactant is expensive. Foam fractionation was investigated for both anionic and cationic surfactant recovery. Chelating agent such as ethylenediaminetetraacetic acid (EDTA) was studied for the separation of heavy metals from surfactant solution. Anionic surfactants bound with heavy metals can be recovered by lowering pH (acidification). In this study, citric acid and imminodiacetic acid (IDA) were applied to release copper from sodium dodecyl sulfate (SDS) micellar solution and compared with EDTA. Precipitation of copper by ferricynide and sodium sulfide were also investigated. As a result, ca. 100 % of copper was released from SDS micellar solution by 5 mM of EDTA and citric acid. And 3.3 mM of ferricyanide formed precipitate with 82.7 % of copper. 5 mM of IDA and sodium sulfide released or formed precipitate 82.5 % and 58.9 % of copper, respectively. Citric acid is harmless to environments and ferricyanide precipitates with Cu easily. Therefore, it is considered that citric acid and ferricyanide have competiveness over a famous chelating agent, EDTA, for the separation of Cu from SDS solution.

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Studies on Secretion of Catecholamines Evoked by Panaxadiol in the Isolated Rabbit Adrenal Gland (파낙스디올의 가토적출부신의 카테콜아민분비 작용에 관한 연구)

  • Lim Dong Yoon;Park Kyu Baik;Kim Kyu Hyung;Choi Cheol Hee;Cha Jong Hee
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.55-62
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    • 1988
  • The effect of panaxadiol (PD). an active component of Korean ginseng saponins on the secretion of catecholamines (CA) from the rabbit adrenal gland and its mode of action were investigated. PD ($400\;{\mu}g$) increased significantly the secretion of CA from the isolated perfused rabbit adrenal gland. PD-induced secretion of CA was reduced markedly by treatment with atropine. CA secretion induced by Ach or PD was potentiated by physostigmine treatment. Chlorisondamine inhibited CA secretion of PD or Ach. Perfusion of PD ($400\;{\mu}g$) for 30 minutes enhanced the secretory activity of CA by Ach. Ouabain weakened the secretory rsponse caused by PD but enhanced the response by Ach. Adenosine treatment resulted in marked increase in CA secretion by PD or Ach. Perfusion with calcium free Krebs solution containing 5 mM EDTA for 30 minutes completely blocked the secretory effect induced by Ach and also weakened that evoked by PD. It is suggested that PD causes the secretion of CA from the rabbit adrenal gland by a calcium dependent exocytotic process. The secretory effect of PD is due to the stimulation of cholinergic muscarinic and nicotinic-receptors present in the adrenal gland partly by direct action on the chromaffin cell.

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