• Title/Summary/Keyword: cytosine

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Effect of Cytosine Analogues on Cytosine Deaminase from Aspergillus fumigatus IFO 5840 (Aspergillus fumigatus IFO 5840의 Cytosine Deaminase에 미치는 Cytosine Analogue의 영향)

  • 김재근
    • The Korean Journal of Food And Nutrition
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    • v.10 no.1
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    • pp.53-59
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    • 1997
  • In this study investigated the effect of cytosine deaminase activity from Aspergillus fumigatus IFO 5840 by cytosine analogues. The results were as follows. The enzyme was strongly inibited by 2-thiouracil, 2-thiocytosine, 6-azacytosine and 2-mercaptopyrimidine. The half inhibitory concentration(HIC) of 2-thiocytosine and 6-azacytosine on cytosine deaminase was 0.80mM and 1.15mM, respectively. The enzyme was inhibited at a certain level by addition of 2-thiocytosine immediately, but was maintained to some extend under the inhibited state by 6-azacytosine in proportion to reaction time. Regardless of kinds of substrate such as cytosine and 5-fluorocytosine, 2-thiocytosine and 6-azacytosine showed action as inhibitors, 2-thiocytosine inhibited cytosine deaminase activity about twice as strong as 6-azacytosine. The enzyme, when cytosine was used as a substrate, was revealed the pattern of competitive inhibition by 2-thiocytosine and 6-azacytosine, The ki value for these compounds was 4.5$\times$10-4M and 1.756$\times$10-3M, respectively. At this point, the Hill coefficient for cytosine, 2-thiocytosine and 6-azacytosine was 1.80, 1.81 and 2.45, respectively.

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Chemical Modification of Intracellular Cytosine Deaminase from Chromobacterium violaceum YK 391

  • Kim, Jung;Kim, Tae-Hyun;Yu, Tae-Shick
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.180-185
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    • 2005
  • Cytosine deaminase (cytosine aminohydrolase, EC 3.5.4.1) stoichiometrically catalyzes the hydrolytic deamination of cytosine and 5-fluorocytosine to uracil and 5-fluorouracil, respectively. Amino acid residues located in or near the active sites of the intracellular cytosine deaminase from chromobacterium violaceum YK 391 were identified by chemical modification studies. The enzymic activity was completely inhibited by chemical modifiers, such as 1mM NBS, chloramine-T, $\rho-CMB,\;\rho-HMB$ and iodine, and was strongly inhibited by 1mM PMSF and pyridoxal 5'-phosphate. This chemical deactivation of the enzymic activity was reversed by a high concentration of cytosine. Furthermore, the deactivation of the enzymic activity by $\rho-CMB$ was also reversed by 1mM cysteine-HCI, DTT and 2-mercaptoethanol. These results suggested that cysteine, tryptophan and methionine residues might be located in or near the active sites of the enzyme, while serine and lysine were indirectly involved in the enzymic activity. The intracellular cytosine deaminase from C violaceum YK 391 was assumed to be a thiol enzyme.

Cytosine Deaminase of Fungus (곰팡이의 Cytosine Deaminase에 관한 연구)

  • ;;Takuo Sakai;Kenzo Tonomura
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.169-174
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    • 1986
  • Cytosine deaminase was partialy purified about 10 fold from the ceil-free extract of Aspergillus fumigatus. The partialy purified enzyme was relatively stable in a pH 5.5 to 8.0, but thermo-unstable. The enzyme activity was found in a pH optimum of 7.0 and temperature optimum of 30 to 35$^{\circ}C$. The activation energy calculated to be 13,240 cal/mol. The apparent Michaelis constants Km for cytosine was found to be 1.53 mM and the molecular weight was determined to be approximately 32,000. The enzyme was strongly inhibited by 0.1 mM of Hg$^^{2+}$, Pb$^{2+}$, Cd$^{2+}$ and Fe$^{2+}$, furthermore inhibited by 1mM of ATP, UTP, o-phenanthroline and p-chloromercuribenzoate.

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Isolation and Identification of Bacterium Producing Extracellular Cytosine deaminase (세포외 Cytosine Deaminase 생산균의 분리 및 동정)

  • 유대식;김대현
    • Microbiology and Biotechnology Letters
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    • v.25 no.1
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    • pp.9-14
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    • 1997
  • A bacterium, strain YK 391 producing extracellular cytosine deaminase, has been isolated from soil sample collected near Taegu City and identified. The strain YK 391 was observed to be a motile Gram-negative rod, and did not produced capsule nor spore. The bacterium produced acid from glucose and trehalose, not from arabinose. Esculine was nto hydrolyzed. The isolate could grow anaerobically at 37$\circ $C, but not at 4$\circ $C. Palmitoleic and palmitic acids comprised over 80% of the fatty acid composition of the strain. The strain. The strain YK 391 was identified as Chromobacterium violaceum YK 391 based on its morphological and physiolohical characteristics, and on the fatty acid composition. The extracellular cytosine deaminase produced by Chromobacterium violaceum YK 391 is believed to be unique because it was active not only on cytosine and 5-fluorocytosine but also on cytidine.

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Preparation and Comparison of Proteus mirabilis and Citrobacter freundii Bacterial Electrodes for the Determination of Cytosine (Cytosine 정량을 위한 Proteus mirabilis와 Citrobacter freundii 박테리아전극의 개발과 그 비교)

  • Gwon Shik Ihn;Bong Weon Kim
    • Journal of the Korean Chemical Society
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    • v.32 no.4
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    • pp.333-341
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    • 1988
  • The bio-electrode for cytosine has been constructed by immobilizing Proteus mirabilis and Citrobacter freundii on an ammonia gas-sensor. Bacteria containing cytosine deaminase convert one molecule of cytosine into one molecule of ammonia. The Proteus mirabilis bacterial electrode showed linear response to cytosine concentration in the $1.0{\times}10^{-3}\;-\;5.0{\times}10^{-2}$M with a slope of 45-48 mV/decade in 0.2 M phospbate buffer solution at pH 8.4. The Citrobacter freundii bacterial electrode showed linear response to cytosine concentration in the $7.0{\times}10^{-5}\;-\;7.0{\times}10^{-3}$M with a slope of 48 mV/decade in 0.05M phosphate buffer solution at pH 7.6. These electrode were investigated for the effects of pH, temperature, buffer solutions, amounts of bacteria, interferences, inorganic salts and lifetime.

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Revelation of Antibacterial Effect Against Cariogenic Bacteria in Combination with 5-Fluorocytosine and Cytosine Deaminase from Chromobacterium violaceum YK 391. (Chromobacterium violaceum YK 391의 세포내 Cytosine Deaminnse와 5-Fluorocytosine의 병용사용이 치아우식 원인세균에 대한 항균성의 발현)

  • Kim, Jung;Jung, Hyuk-Jun;Yoo, Dae-Sik
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.292-296
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    • 2003
  • The antibacterial effect against cariogenic bacteria was evaluated in combination of 5-FC and intracellular cytosine deaminase from Chromobacterium violaceum YK 391. While S. mutans, L. parabuchneri and A. naeslundii showed antibacterial effect against 10 mM of 5-FU, S. intermedius, S. mitis, S. agalactiae, L. lactis, A. israelii, A. viscosus don't caused antibacterial effect. The addition of the cytosine deaminase and 10 mM of 5-FC to S. mutans, S. sanguis, L. brevis, L. parabuchneri, L. oris and A. naeslundii caused weakly antibacterial effect. S. sanguis caused weakly antibacterial effect against 10 mM of 5-FC. These results suggested that combination of the cytosine deaminase and 5-FC was showed the possibility to precautionary measures of dental caries.

The Electronic Structure of Interaction Platinum(Ⅱ) with DNA bases, Adenine, Guanine, and Cytosine (Platinum(Ⅱ) Complex와 DNA bases인 Adenine, Guanine 그리고 Cytosine의 Interaction에 대한 전자구조)

  • Kim, Ui Rak;Kim, Sang Hae;Edward A. Boudreaux
    • Journal of the Korean Chemical Society
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    • v.34 no.6
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    • pp.539-547
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    • 1990
  • Molecular Orbital calculations using the SC-MEH method have been carried out for the interaction of Adenine, Guanine and Cytosine as DNA base and diaminecytosineplatinum(DCP) in various conformations. The results showed that the order of DCP binding to the DNA bases was guanine > adenine > cytosine and the stabilization energy of cis-isomer was larger than that of trans-isomer in the adenine-DCP complexes system. Furthermore, platinum(II) binding to DNA bases markedly gives rise to change of atomic charge in DNA bases ring, which can explain anti-tumor activity of platinum complex.

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Purification and Properties of Intracellular Cytosine Deaminase from Chromobacterium violaceum YK 391

  • KIM , JUNG;YU, TAE-SHICK
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1182-1189
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    • 2004
  • Cytosine deaminase (cytosine aminohydrolase, EC 3.5.4.1) stoichiometrically catalyzes the hydrolytic deamination of cytosine and 5-fluorocytosine to uracil and 5-fluorouracil, respectively. The intracellular cytosine deaminase from Chromobacterium violaceum YK 391 was purified to apparent homogeneity with 272.9-fold purification with an overall yield of $13.8\%$. The enzyme consisted of dimeric polypeptides of 63 kDa, and the total molecular mass was calculated to be approximately 126 kDa. Besides cytosine, the enzyme deaminated 5-fluorocytosine, cytidine, 6-azacytosine, and 5-methylcytosine, but not 5-azacytosine. Optimum pH and temperature for the enzyme reaction were 7.5 and $30^{\circ}C$, respectively. The enzyme was stable at pH 6.0 to 8.0, and at 30T for a week. About $70\%$ of the enzyme activity was retained at $60^{\circ}C$ for 5 min. The apparent $K_{m}$ values for cytosine, 5-fluorocytosine, and 5-methylcytosine were calculated to be 0.38 mM, 0.87 mM, and 2.32 mM, respectively. The enzyme activity was strongly inhibited by 1 mM $Hg^{2+},\;Zn^{2+},\;Cu^{2+},\;Pb^{2+},\;and\;Fe^{3+}$, and by o-phenanthroline, $\alpha,\;{\alpha}'$-dipyridyl, p-choromercuribenzoate, N-bromosuccinimide, and cWoramine­T. In addition, the enzyme activity was strongly inhibited by I mM 2-thiouracil, and weakly inhibited by 2-thiocytosine, or 5-azacytosine. Finally, intracellular and extracellular cytosine deaminases from Chromobacterium violaceum YK 391 were found to have a different optimum temperature, apparent $K_{m}$ value, and molecular mass.

Optimal Conditions for the Production of Intracellular Cytosine Deaminase from Chromobacterium violaceum YK 391. (Chromobacterium violaceum YK 391의 세포내 Cytosine Deaminase의 생성 최적조건)

  • Kim, Jung;Kim, Hyun-Soo;Yoo, Dae-Sik
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.367-372
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    • 2002
  • Cytosine deaminase (cytosine aminohydrolase, EC 3.5.4.1) stoichiometrically catalyzes the hydrolytic deamination of cytosine and 5-fluorocytosine to uracil and 5-fluorouracil, respectively. Optimal medium compositions for production of cytosine deaminase from Chromobacterium violaceum YK 391 were 0.75% soluble starch, 1.5% peptone, 0.1% meat extract, 0.1% yeast extract, 0.01% NaCl, 0.01% $MgCl_2{\cdot}7H_2O$ and 0.05% $K_2HPO_4$. The optimal pH of medium and incubation temperature were 7.0 and $30^{\circ}C$, respectively. C. violaceum reached stationary phase after 30 hr, and produced a maximum cytosine deaminase (120 units/ml) after 72 h in batch culture.

Bacillus stearothermophilus 에서 부분 정제한 Cytosine Deaminase 의 특성

  • 장영채;이경형;김성영;조윤래;김종규
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.305-309
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    • 1992
  • Cytosine deaminase (EC 3.5.4.1) from BaciNus stc~urorhermophilus was partially purified 7.2-fold with an overall yield of 52.7%. The partially purified enzyme deiiminated cytosine only.but not 5-methylcytosine and 5-fluorocytosine. The apparent Michaclis constant. Km valuefor cytosine was 5.9 mM. The enzyme was relatively stable in the range of pH 4.0 to 7.0.furthermore extremely thermo-stable : more than 75'X) of the activity was remained afterheating at 80$^{\circ}$C for I0 min at pH 6.5. The enzyme had a pH optimum at around pH7.0 to 7.5. and temperature optimum at 35 to 31$^{\circ}$C. And the activation energ (En value)determined from an Arrhenius plot was 26 Kcal/mol. The enzyme activity was stronglyinhibited by heavy metal ions such as Cd", Hg". Cut' at 1 mM, anJ by o-phenanthroline,and p-chloromcrcuribcnzoate at I mM. But the enrymc activity was activatetl increased byGMP, and CMP at 1 mM.ased by GMP, and CMP at 1 mM.

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