Characterization of Acid Phosphatase from Welsh Onion

파의 Acid Phosphatase의 특성

Acid phosphatase (EC 3.1.3.2) from Welsh onion was partially purified by Sephacryl S-200 gel filtration and CM-Sepharose CL-6B ion exchange chromatography. The optimum pH and temperature of acid phosphatase from green onion were pH 5.5 and $60^{\circ}C$, respectively. The enzyme was the most stable at pH 6.0 and unstable above pH 9.0. The activation energy of the enzyme was determined to be 4.86kcal/mole. The enzyme utilized p-nitrophenyl phosphate most as a best substrate among tested possible substrates, while 5'-GMP and 5'-IMP were poor substrates for the enzyme. $K_{m.app.}$ of the enzyme with p-nitrophenyl phosphate as a substrate was identified as 0.87mM. Among metal ions and inhibitors tested, $Cr^{+++},\;Zn^{++},\;Cu^{++}$, molybdate and metavanadate ions inhibited the enzyme reaction drastically.

파로부터 acid phosphatase를 Sephacryl S-200 gel filtration과 CM-Sepharose CL-6B ion exchange chromatography를 이용하여 부분정제하였다. p-nitrophenyl phosphate를 기질로 사용했을 경우에 최적pH는 5.5, 최적온도는 $60^{\circ}C$였다. 효소의 활성화 에너지는 4.86kcal/mole이었다. 효소는 pH 6.0에서 가장 안정하였으며, $50^{\circ}C$ 이하에서 대체로 안정하였다. 효소는 p-nitrophenyl phosphate를 기질로 가장 잘 이용하였으며, 5'-IMP와 5'-GMP는 기질로 거의 이용하지 못하였다. 효소는 p-nitrophenyl phosphate를 기질로 했을 경우에 $K_{m.app.}$값이 0.87mM이었다 $Cr^{+++},\;Zn^{++},\;Cu^{++}$이온은 효소의 활성을 저해하였으며, 또한 molybdate와 metavanadate 이온이 효소의 활성을 저해하였다. 그리고, NaCl의 농도가 높을수록 효소의 활성을 저해하였다.