Optimal Conditions for the Production of Intracellular Cytosine Deaminase from Chromobacterium violaceum YK 391.

Chromobacterium violaceum YK 391의 세포내 Cytosine Deaminase의 생성 최적조건

  • 김정 (수원여자대학 치위생과) ;
  • 김현수 (계명대학교 자연과학대학 미생물학과) ;
  • 유대식 (계명대학교 자연과학대학 미생물학과)
  • Published : 2002.12.01

Abstract

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.

본 연구에서는 cytosine deaminase(CODase)의 활성이 비교적 높은 Chromobacterium violaceum YK 391의 생육과 세포내 CODase의 최적 배양조건을 규명한 결과, soluble starch 0.75%, peptone 1.5%, yeast extract 0.1%, meat extract 0.1%, NaCl 0.01%, $K_2HPO_4$ 0.05%였다. 배지의 초기 pH 6.5에서 7.5까지 생육과 더불어 효소생성이 가장 양호했지만 산성 혹은 염기성 쪽으로 갈수록 균의 증식의 둔화와 더불어 효소생성 정도도 낮아졌다. 효소 생성에 미치는 배양 온도의 영향을 검토한 결과, 균의 생육은 저온보다는 $28^{\circ}C$에서 $30^{\circ}C$의 온도에서 양호하였으며, 40도씨 이상의 고온에서는 생육 및 효소 생성이 점차적으로 낮아졌고 균의 증식과 배양일수에 따른 효소활성에 있어서 균은 12시간의 유도기를 거쳐 배양 72시간 이후에 최대 생육도를 나타냈으며 균의 증식과 효소 생성 정도는 거의 일치하는 양상을 나타냈다.

Keywords

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