Immobilization of Cellulases from Fomitopsis pinicola and Their Changes of Enzymatic Characteristics

Formiptosis pinicola KMJ812에 의해 생산된 조효소액은 섬유소 분해효소 복합체로서 매우 활성이 높으며 특히 ${\beta}$ -glucosidase의 활성이 높아 포도당 전환수율이 높다. 본 연구에서는 F. pinicola KMJ812 생산 cellulase를 고정화에 따른 효소특성의 변화와 고정화 효소의 재사용에 따른 효소의 불활성 정도를 관찰하였다. 담체는 고정화 수율이 cellulase활성 61.7%와 ${\beta}$ -glucosidase활성 64.4%로 우수한 Duolite A568로 선정하였다. 고정화 효소의 최적반응온도는 cellulase와 ${\beta}$ -glucosidase 활성의 경우 모두 $55^{\circ}C$ 로서 수용성효소보다 높았으며, 최적 pH는 cellulase활성은 4.0이었고, ${\beta}$ -glucosidase활성은 4.5로 cellulase활성의 경우에서만 수용성효소와 비교하여 약간 염기성으로 변화하였다. 본 고정화 효소는 $50^{\circ}C$ 에서 72시간 후에 98%의 활성을 유지하고 있었으며, $50^{\circ}C$ 에서 8회 사용 후에 50%정도의 잔존활성을 나타내었다. Cellulase from Formiptosis pinicola KMJ812 is an efficient cellulose degradation enzyme complex, especially with a high ${\beta}$ -glucosidase activity. In this study, the change in enzymatic characteristics by immobilization and the reduction of immobilized enzyme activity by repeated usages were evaluated using cellulases from F. pinicola KMJ812. Among tested four resins, Duolite A568 resin had the best enzyme activity yield with 61.7% cellulase activity and 64.4% ${\beta}$ - glucosidase activity during the cellulase immobilization. The best reaction temperature was $55^{\circ}C$ for both cellulase and ${\beta}$ -glucosidase activities which were higher than the unimmobilized soluble cellulases. The best reaction pH was 4.0 for cellulase activity which was a little more basic than a soluble form and 4.5 for ${\beta}$ -glucosidase activity. The immobilized cellulase activity was remained 98% of the beginning activity after 72 h incubation at $50^{\circ}C$ and 50% of the beginning activity after eight times usage at $50^{\circ}C$ .