Cyclodextrin Glucanotransferase와 Cyclodextrinase를 생산하는 Bacillus 속 세균의 분리와 그 효소들의 특성

A bacterium producing Cyclodextrin Glucanotransferase (CGTase) and Cyclodextrinase (CDase) was isolated from soil, and named as Bacillus stearothermophilus KJ16. The growth of the isolated strain occurred in two steps, and syntheses of CGTase and CDase were dependedt on the growth cycle of the cell. CGTase was constitutively synthesized during the 1st growing phase, while CDase was synthesized inducibly during the 2nd growing phase. When the midium pH was controlled at 7.0 the maximum enzyme activities of CGTase and CDase were increased by 12-fold (1300 mU/ml) and 2-fold (225 mU/ml), respectively, compared with the pH-uncontrolled batch culture. The CGTase of the isolate converted soluble starch to CDs with the ratio of $\alpha$-CD:$\beta$-CD:$\gamma$-CD=42:46:12 at $55^{\circ}C$.The optimal pH and temperature of CGTase were 6.0 and $60^{\circ}C$, respectively and the optimal pH and temperature of CDase were 6.0 and $55^{\circ}C$. The molecular weights of the purified CGTase and CDase were estimated to be 65, 000 and 68, 000 dalton, respectively. Among several substrates, $\gamma$-CD was most rapidly hydrolyzed by the purified CDase.