참나물로부터 추출한 polyphenol oxidase의 부분정제를 $(NH_4)_2SO_4$, 처리 및 Sephadex G-150 column chromatography에 의해 분리하였다. Polyphenoloxidase의 최적 pH와 최적온도는 7.5와$30^{circ}C$였으며, pH 6.5에서 $70^{circ}C$에서 30분간 처리시와 $80^{circ}C$에서 5분간 처리시 완전 실활하였다. Polyphenol oxidase는 ascorbic acid, glutathione, potassium cyanide(0.1mM)에 의해 불활성화되었으며 L-cysteine, potassium cyanide, ascorbic acid, glutathione(0.5, 1mM)에 의해서는 완전 실활되었다. Catechol과 3,4-dihydroxytoluene의 기질은 높은 특이성을 나타낸 반면 pyrogallol, dopamine, DL-dopa의 기질은 강하게 활성을 억제하였다. Polyphenol oxidase의 $K_m$ 값은 86.5mM이었다.
Three polyphenol oxidase(polyphenol oxidase I, II and III ) were isolated from the crude extract of a Spuriopimpinella bracycarpa by $(NH_4)_2SO_4$ precipitation and subsequent Sephadex G-150 chromatography. The final preparation thus obtained showed three peaks of enzyme activity. Optimum pH and temperature for the activity of polyphenol oxidase were 7.5 and $30^{circ}C$, respectively. The enzyme was completely inactivated when i4 was treated at$70^{circ}C$ for 30min and at $80^{circ}C$ for 5min at pH 6.5. The enzyme was partially inactivated by ascorbic acid, glutathione and potassium cyanide (0.1mM), and was completely inhibited by L-cysteine, ascorbic acid, glutathione and potassium cyanide(0.5 and 1.0mM). The enzyme has good activity on catechol and 3,4-dihydroxytoluene but was strongly inactivated on pyrogallol, dopamine and DL-dopa. The Michaelis cons4ant of the enzyme was 86.5mM with catechol as a substrate.
