논문 상세보기
pp.176-180
열에 안정한 단백질 분해효소를 탐색하고자 토양에서 고온성 방선균을 분리 선별하여 여기서 생산되는 효소를 정제하여 특성을 조사하였다. 본 고온성 방선균의 효소생산 조건은 0.5% soluble starch, 0.5% yeast extract, 0.1% K_2HPO_4, 0.05% CaCl_2, initial pH 8.0, 50℃에서 48시간 배양하였다. 정제 방법은 ammonium sulfate의 염석, DEAE HPLC, GPC HPLC 순으로 정제하였다. 본 효소의 최적 pH와 온도는 7.5와 60℃이였으며, 안정성은 pH 6.5∼8.5사이에서 안정하였으나 80℃에서 10분간 열처리에서는 90% 실활시켰다. Ag^+와 Cu^2+이온은 효소활성을 저해시켰다.
Microbial protease has been interesting due to the biological roles in the producing microorganism. A thermophilic Actinomyces producing protease was isolated from soil. The optimal medium composition and culture conditions for maximum protease production was as follows 0.5% soluble starch, 0.5% yeast extract, 0.1% K_2HPO_4, 0.05% CaCl_2, initial pH 8.0 at 50℃ for 48hours. The protease was purified by the procedure of ammonium sulfate precipitation, anion exchange chromatography(LC), DEAE high performance liquid chromatography precipitation, anion exchange chromatography(LD), DEAE high performance liquid chromatography and GPC HPLC. The purification fold of the purified enzyme was increased about 22.6. The optimal pH and temperature for reaction of the purified enzyme were 7.5 and 60℃. The purified enzyme was stable for the pH range from 6.0 to 8.5, but was unstable when treated at 80℃ for 10 minutes. The activity of the enzyme was inhibited by Ag^+ and Cu^2+.
