Aspergillus nidulance가 생산하는 Naringinase에 관한 연구 -제 3 보 고정화(固定化) Naringinase의 제조 및 그 성질-

Studies on Naringinase Produced from Aspergillus nidulance -Part III. Preparation and Properties of Immobilized Naringinase-

Aspergillus nidulans가 생산하는 naringinase의 acrylamide gel을 이용한 고정화(固定化)의 조건과 고정화효소의 성질 및 column reactor의 반응조건을 연구 검토하였는 바 그 결과를 요약하면 다음과 같다. 1. 고정화효소를 제조하는데 acrylamide의 농도는 10%가 적합하였으며, N.N'-methylene bis acrylamide의 농도는 0.5%이상에서 영향이 없었다. 이 조건에서 효소농도는 126 unit/ml가 적합하였다. 2. 고정화효소의 최적반응 pH는 5.0으로서 수용성 효소와 동일하였으며, 최적반응온도는 $50^{\circ}C$로서 수용성 효소에 비해 $10^{\circ}C$상승하였다. 3. Naringinase는 acrylamide gel에 고정시키므로서 pH안정성과 열안정성은 현저히 향상되었다. 4. 고정화효소의 활성화에너지는 수용성 효소보다 약 1/2적은 13.01 kcal/mole이였으며, apparent Km값은 $7.23{\times}10^{-3}$ mole이였다.

Naringinase produced from Aspergillus nidulans was immobilized in acrylamide gel by the entrapping method and its characteristics were studied. Optimum acrylamide concentration was 10%, but N.N'-methylene bisacrylamide concentration had no influence on the final enzyme gel activity. The suitable amount of enzyme dissolved in the polymerization reaction mixture was 126 units/ml. Optimum pH of immobilized enzyme was 5.0 which was the same as that of free enzyme. However, immobilized enzyme showed a higher optumum reaction temperature, markedly increased pH and temperature stability. In a packed-column reactor, the observed reaction rate was increased proportionally to flow rate up to 5ml/min., but independent above 6ml/min.. Activation energy of the immobilized enzyme was 13.01 Kcal/mole, and the energy required for the thermal inactivation was 39.4 Kcal/mole. The apparent Km for 100 mesh gel was $7.23{\times}10^{-3}$ mole.