Continuous Production of Transglucosylated Steviosides Using Immobilized Cyclodextrin Glucanotransferase

고정화 Cyclodextrin Glucanotransferase에 의한 당전이 스테비오사이드의 연속생산

In order to produce transglucosylated steviosides continuously, some types of bioreactors were investigated with cyclodextrin glucanotransferase immobilized on a high porous anion exchange resin, Diaion HPA75. Among the bioreactors, the packed-bed reactor (PBR) showed the highest specific productivity. The effect of linear velocity in a PBR on the stevioside conversion was not significant in the range of $10{\sim}60\;cm/hr$ at the same space velocity of $1.2\;hr^(-1)$. When the space velocity of bioreactor was varied from 0.2 to $1.1\;hr^{-1}$, the optimal velocity of substrate solution was determined as $0.7\;hr^(-1)$. The stevioside conversion of more than 70% was maintained during 20 days in the continuous operation, if about 20% of immobilized enzyme was replaced in the top of reactor after two weeks operation as the one of the control methods in bioreactor. The specific production, which refers to as the amount of commercially valuable transglucosylated steviosides produced by a unit amount of immobilized cyclodextrin glucanotransferase, was found to be ca. 150g product/g immobilized enzyme.

고정화 CGTase를 이용하여 당전이 스테비오사이드를 연속적으로 제조하기 위하여 몇가지 생물반응기의 적용가능성을 조사하였다. 연속형 반응기로는 packed-bed reactor(PBR)가 적합하였다. 이때 specific productivity는 0.4 g/hr-g immobilized enzyme으로 회분식반응기의 50% 정도이나 연속생산이 가능한 장점이 있었다. PBR의 운전조건이고 $10{\sim}60\;cm/hr$의 선속도 범위내에서 스테비오사이드 전환률 70% 이상을 보이는 기질의 유속은 공간속도로 $0.5{\sim}0.7\;hr^{-1}$이었다. 고정화 효소의 운전 안정성은 $47^{\circ}C$에서 30%의 기질로 2주 일간 연속 사용이 가능하고, 반응기의 control방법으로 반응기 상부에서 20%의 효소를 교체하면 20일 이상 전환률이 유지되었다.