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자료유형
학술저널
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한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제14권 제6호
발행연도
2004.1
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1,182 - 1,189 (8page)

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Cytosine deaminase (cytosine aminohydrolase, EC3.5.4.1) stoichiometrically catalyzes the hydrolytic deaminationof cytosine and 5-fluorocytosine to uracil and 5-fluorouracil,respectively. The intracellular cytosine deaminase fromChromobacterium violaceum YK 391 was purified toapparent homogeneity with 272.9-fold purification with anoverall yield of 13.8%. The enzyme consisted of dimericpolypeptides of 63 kDa, and the total molecular mass wascalculated to be approximately 126 kDa. Besides cytosine, theenzyme deaminated 5-fluorocytosine, cytidine, 6-azacytosine,and 5-methylcytosine, but not 5-azacytosine. Optimum pHand temperature for the enzyme reaction were 7.5 and 30oC,respectively. The enzyme was stable at pH 6.0 to 8.0, and at30oC for a week. About 70% of the enzyme activity wasretained at 60oC for 5 min. The apparent Km values forcytosine, 5-fluorocytosine, and 5-methylcytosine were calculatedto be 0.38 mM, 0.87 mM, and 2.32 mM, respectively. Theenzyme activity was strongly inhibited by 1 mM Hg2+, Zn2+,Cu2+, Pb2+, and Fe3+, and by o-phenanthroline, α,α'-dipyridyl,ρ-chloromercuribenzoate, N-bromosuccinimide, and chloramine-T. In addition, the enzyme activity was strongly inhibited by1 mM 2-thiouracil, and weakly inhibited by 2-thiocytosine, or5-azacytosine. Finally, intracellular and extracellular cytosinedeaminases from Chromobacterium violaceum YK 391 werefound to have a different optimum temperature, apparent Kmvalue, and molecular mass.

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