초록
뽕나무종자(Morus Lhou Koidz)를 28$^{\circ}C$의 암소에서 5일간 발아시키면서 매일 hormone을 농도별로 처리한 후 protease 활성의 변화를 각각 비교하였다. 또 효소활성이 가장 높았던 4일째 시료로부터 protease를 추출하여 부분정제하고 그 효소의 특성을 조사하였다. 그 결과는 다음과 같다. 1. Hormone 처리구의 발아종자는 효소활성이 GA3가 가장 높았고 그 다음이 zeatin과 kinetin이었으며 hormone 혼합구는 GA3 10$\mu$M에 zeatin 10$\mu$M과 kinetin 10$\mu$M을 혼합한 구가 가장 활성이 높았다. 2. 효소활성은 10ml 주입한 한천배지구가 대조구보다 4일째 약 14% 증가하였다. 3. 뽕나무씨앗으로부터 추출한 protease는 DEAE-Toyopearl 650M, Butyl-Toyopearl, Hydrozyl apetite 및 Toyopearl HW55M 방법으로 조효소의 313배로 정제하였다. 정제한 후 효소의 비활성이 175unit/mg였다. 4. 효소반응이 최적 pH는 5.0, 최적온도는 37$^{\circ}C$였다. 효소는 37$^{\circ}C$이하에서는 안정하였으나 52$^{\circ}C$에서는 10분간 처리로 약 50% 감소하였다. 5. 효소활성을 수은, 철, 아연 및 구리 이온에 의하여 활성이 억제된 반면 Mg, Cr 및 A1은 촉진되었다. 또한 효소활성은 azocasein을 기질로 했을 때 Km값은 0.89mM이었다.
The activity change of mulberry seeds protease was compared during germination for 5 days at 28$^{\circ}C$ in the dark place after daily hormone injection of different concentration. The protease from germinated mulberry seeds for 4 days was partially purified and the enzyme characteristics was investigated. The protease activity of mulberry seeds treated by hormone was highest with 10 $\mu$m GA3 followed by 10 $\mu$M zeatin and 10 $\mu$M kinetin. The protease activity of mulberry seeds was increased by 14% with 10ml agar culture that control at 4th day of germination. The protease from mulberry seeds was purified 313 fold by DEAE-Toyo-pearl 650M, Butyl-Toyopearl, Hydrozylapatite and Toyopearl HW 55M. After purification, the specific activity of the enzyme was 175 units/mg. Optimum pH and temperature of protease from mulberry seeds was 5.0 and 37$^{\circ}C$, respectively. The protease was stable below 37$^{\circ}C$ and the enzyme activity was decreased by 50%, when incubated at 52$^{\circ}C$ for 10minutes. The protease activity of mulberry seeds was inhibited by metal ions such as mercury, iron, zinc, copper, but activited by magnesium, choromium, aluminium ions. The Km value of the protease was 0.89mM with azocasein as a subscribe.