Abstract
Asp. usumii IAM 2185 was selected as a strain producing the powerful raw starch digesting glucoamylase. The optimum initial pH, the optimum temperature and the optimum cultural time for the enzyme production on wheat bran medium were pH 6-8,25-$30^{\circ}C$ and 72 hrs, respectively. The addition of ammonium nitrate and albumin on wheat bran medium, respectively, increase slightly the enzyme production. The enzyme was purified by ammonium sulfate fractionation, CM-cellulose and DEAE-cellulose column chromatography. The specific activity of the purified enzyme was 34.3 U/mg protein and the yield of enzyme activity was 10.3%. The purified enzyme showed a single band on polyacrylamide disc gel electrophoresis and its molecular weight was estimated to be 67,000 by SDS polyacrylamide disc gel electrophoresis. The isoelectric point for the purified enzyme was pR 3.7. The optimum temperature and optimum pH were $60^{\circ}C$and pH 3.0 and the purified enzyme was stable in the pH range of 1.0-11.0. The purified enzyme was stable below $50^{\circ}C$ and its thermostability was greatly increased by the addition of $Ca^{2+}$. The purified enzyme showed a high hydrolysis rate on various raw starches such as corn, rice, yam, arrow root, sweet potato and glutinous rice.
생전분 분해력이 강력한 glucoamylase를 생산하는 균주로서 Asp.usamii IAM 2185를 선정하였다. 밀기울배지에서의 효소생산의 최적 initial pH는 6.0-8.0, 최적 배양온도는 25-$30^{\circ}C$, 최적 배양시간은 72시간이고, 밀기울배지에 ammonuim nitrate와 albumin의 첨가는 효소의 생산을 약간 증가시켰다. 황산암모늄분획, CM-cellulose와 DEAE-cellulose column chromatography에 의하여 효소를 정제하였고, 정제효소의 specific activity는 34.3U/mg.protein, 수율은 10.3 이었다.