Properties of an Extracellular 5-Fluorocytosine Deaminase

세포외 5-Fluorocytosine Deaminase의 특성

- Some properties of an extracellular cytosine deaminase excreted from an isolate from soil samples were examined after 20~80%' ammonium sulfate fractionation. The enzyme catalyzed the conversion of cytosine and 5-fluorocytosine into uracil and 5-fluorouracil by substrate specificity, respectively. The optimum temperature and storage time on the stability of the enzvme preparation were below $50^{\circ}C$ keeping above 90% of the residual activity and near 4 days keeping above 80% of the residual one in Tris-HCI buffer. The maximum activity was also obtained at 8.0 in pH and 37'C in temperature. The pHs and temperatures for enzyme activity ranged from 8.0~8.5 and from 37~$45^{\circ}C$. respectively. The presence of $Ag^-,Hg^{2-}, Zn^{2-}, Cu^{2-}, Sn^{2-}, \; or\; Pb^{2-}$ in the reaction mixture resulted in the marked inhibition in enzyme activity, but 1 mM of $K^+, Fe^[3+}, Mg^{2+}, \; or \; Na^+$. slightly increased the activity. The enzyme preparation was heavily affected by most of inhibitors tested such as 1 mM of EIITA, NaCN and pentachlorophenol, and completely inactivated by p-CMB and TCA of 1 mM, or 10 mM.

토양 분리 균주로부터 얻은 배양액을 20-80 황산암모늄으로 분획한 후 세포의 cytosine deaminase의 성질을 검토하였다. 이 효소는 cytosine과 5-fluorocytosine(5-FC)에 기질 특이성을 가짐으로서 각각 uracil과 5-fluorouracil(5-FU)로 전환을 촉매하였다. 효소안정성에 대한 온도와 보존기간은 tris-HC1 완충용액에서 검토한 결과 최적온도는 $50^{\circ}C$ 부근이하에서 90 이상의 잔존활성을, 보존시간은 4일정도에서 80 이상의 잔존활성을 유지하였다. 최대 pH 8.0과 $37^{\circ}C$의 온도에서 나타났다. 활성에 적당한 pH는 8.0~8.5였고, 온도는 37~$45^{\circ}C$의 범위였다.