The production of Alkaline Protease by Aspergillus fumigatus and Purification of Enzyme

Aspergillus fumigatus에 의한 Alkaline Protease의 생산과 정제

The alkaline protease producing mold isolated from and identified as Aspergillus fumigatus. It was found that the production of alkaline protease reach to maximum was cultured for 3 days at $30^{\circ}C$. The enzyme was purified 86.13 fold and yield of the enzyme purification was 6.4%, The purification procedure include ammonium sulfate treatment, gelfiltration on Sephadex G-25, G-75, G-150 and DEAE-cellulose ion-exchange chromatography. When the purified enzyme was applied sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight was estimated about 63000. This enzyme composed 17 amino acids and main amino acids of this enzyme were glycine and glutamic acid.

Alkaline protease 생성능이 강한 Aspergillus fumigatus을 토양에서 분리동정하고, 효소생산조건을 구명한 결과 $30^{\circ}C$에서 3일간 배양하였을 때 최고활성을 나타냈으며 생산된 조효소를 황산암모늄염석, Sephadex G-25, G-150 gel filteration과 DEAE-Cellulose컬럼 chromatography로 정제하여 수율 6.4%, 정제정도 86.13배의 효소를 얻었고 polyacrylamide gel 전기영동에 의해 단일밴드인 것을 확인하였다. 정제효소의 분자량은 63000정도 였으며, 17종의 아미노산으로 구성되어 있고, 그 중 glycine과 glutamic acid가 가장 많고 methionine과 cystine이 가장 적었다.