고정화 글루코아밀라제의 성질과 응용

Characteristics and Applications of Immobilized Glucoamylase

Glucoamylase를 $ZrO_2$로 피복된 96% porous glass에 azo-linkage를 형성시켜 결합하게 한후, 2.5% glutaraldehyde로 처리하여 효소를 고정화시켰다. 효소기질로는 용해도가 높고 점도가 낮은 30% enzyme thinned cornstarch (dextrose equivalent 값 : 24)를 사용하여 plug flow-column reactor에서 연속반응시켰다. 반응 최적 pH는 수용성효소의 5.0보다 alkaline 쪽으로 기울어져 7.0으로 나타났고, 고정화반응에 따라 열안정성이 높아지고 $40{\sim}60^{\circ}C$에서 최적 온도범위를 가리키며, Km값은 수용성 효소의 1.25mM보다 낮은 1.04mM값을 보여 주었다. 따라서, pH 7.0, $45^{\circ}C$에서 160시간 동안 corn starch를 기질로 효소반응을 시켜 glucose 90.3%, maltose 8.0%인 DE값 94.0인 전분당분해산물을 획득할 수 있었다.

Glucoamylases catalyze a stepwise hydrolysis of starch with the production of glucose. In order to make an efficient conversion of starch into glucose, glucoamylases prepared from Rhizopus spp. (Sigma Co.) were attached to a porous glass and immobilized by glutaraldehyde-induced crosslinking. The porous glass used in this study was $ZrO_2$ coated, $40{\sim}80$ mesh, 550 A pore diameter. Using the forgoing glass, we could couple as much as 50mg of protein per gram of carrier. Substrate for the glucoamylase was an enzyrne-modified thin-toiling 30% cornstarch solution used where greater solubility and low viscosity are desired. Immobilized glucoamylase had an optimum pH 7.0 to the alkaline side of soluble enzyme. Km values of immobilized and soluble enzyme were 1.04 mM and 1.25mM, respectively. The thermal stability of glucoamylase was increased by immobilization and the immobilized enzyme showed an optimum temperature at $40{\sim}60^{\circ}C$. The continuous conversion of cornstarch to glucose by use of immobilized glucoamylase resulted in the production of a more than 90 DE product.