Properties of Extracellular Cytosine Deaminase from Arthrobacter sp. JH-13

Arthrobacter sp.JH-13이 생산하는 세포외 Cytosine Deaminase의 성질

Some properties of an extracellular cytosine deaminase produced from Arthrobacter sp.JH-13 were examined after 20-80% of ammonium sulfate fractionation. Among some substrates, this enzyme utilized cytosine and 5-fluorocytosine as a substrate. The optimum pH and temperature for the activity of this enzyme were found to be near 8.0 and $40^{\circ}C$, respectively. The ensyme was more stable in 0.2M of Tris-HCl buffer than 0.2M of potassium phosphate buffer. The enzyme was generally stable below $50^{\circ}C$, but inactivated completely at $70^{\circ}C$. 1mM of $Fe^{3+},\;K^+\;and\;Na^+$ increased the enzyme activity, but 0.01mM of $Co^{2+},\;Cu^{2+},\;Ni^{2+},\;Hg^{2+},\;Ag^{2+},\;Zn^{2+},\;Ba^{2+},\;and\;Mg^{2+}$ markedly inactivated the enzyme activity. 0.1mM of p-chloromercuribenzoate, trichloroacetic acid, and N-ethylmaleimide compleyely inhibited the enzyme activity, but 0.1mM of 2-mercaptoethanol slightly increased the enzyme activity.

20-80%로 황산암모늄 분획한 효소액을 사용하여 Arthrobacter sp. JH-13균주가 생산하는 세포외 cytosme deaminase의 성질을 검토하였다. 검토한 기질 중, 본 효소는 cytosine과 5- f1uorocytosine을 기질로 이용하였으며, 효소의 활성에 대한 최적 pH는 8.0부근이었고, 최적 온도는 $40^{\circ}C$부근으로 나타났다. 본 효소는 0.2M의 p potassium phosphate 완충액 (pH 8.0) 보다 0.2M의 tris-HCl 완충액 (pH 8. 0) 에서 더욱 안정하였다. 온도에 대한 안정성을 검토한 결과. $50^{\circ}C$ 부근까지는 대체로 안정하였으나, $70^{\circ}C$에서는 완전히 활성을 잃었다. 또한 ImM의 $Fe^{3+},\;K^+\;Na^+$ 이온은 효소의 활성을 증가시켰으나 0.01mM의 $Co^{2+},\;Cu^{2+},\;Ni^{2+},\;Hg^{2+},\;Ag^{2+},\;Zn^{2+},\;Ba^{2+},\;Mg^{2+}$ 이온들은 효소의 활성올 강력하게 저해하였다. O.lmM의 p-mercuribenzoate, trichloroacetic acid, N-ethylmaleim mide등은 효소의 활성을 완전히 저해하였으며. O.lmM의 2-mercaptoethanol은 효소의 활성올 약간 증가시키는 것으로 나타났다.