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Journal of microbiology and biotechnology v.14 no.6, 2004년, pp.1182 - 1189   피인용횟수: 1
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Purification and Properties of Intracellular Cytosine Deaminase from Chromobacterium violaceum YK 391

KIM , JUNG    (Department of Dental Hygiene, Suwon Womens College   ); YU, TAE-SHICK    (Department of Microbiology, Keimyung University  );
  • 초록

    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.


  • 주제어

    Cytosine deaminase .   Chromobacterium violaceum YK 391 .   5-fluorocytosine.  

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  • 이 논문을 인용한 문헌 (1)

    1. 2005. "" Biotechnology and bioprocess engineering, 10(3): 180~185     

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