Streptomyces 속 균주가 생성하는 Trypsin Inhibitor (제1보) 균의 분리 및 저해물질의 정제

Trypsin Inhibitor from Streptomyces sp. ( Part 1) Isolation of microorganism and purification of the inhibitor

Trypsin에 대한 강한 저해물질을 생성하는 Streptomyces속 균주 AS-707을 토양으로부터 얻어 그 배양액에서 Trypsin inhibitor를 분리정제하여 저해물질의 안정성과 여러가지의 protease에 대한 저해성 여부를 검토한 결과는 다음과 같다. 배양액을 Amberlite IRC-50에 흡착 methanol추출. 2차 Amberlite IRC-50, CM-cellulosecolumn chromatography로 정제하여 active amorphous powder를 얻었는데 이 때의 비율은 26%였다. 분리정제된 물질은 trypsin 이외에 papain, $\alpha$-chymotrypsin, Azotobacter vineiandi protease와 Bacillus subtilis protease 등에 대해서도 저해작용을 나타내었으며, 안정성은 비교적 커서 10$0^{\circ}C$에서 120분간 가열해도 잔존활성이 약90%였으며, pH처리에 대해서는 37$^{\circ}C$에서 처리하면 산에서 Alkali에 걸치는 대단히 넓은 pH범위 (pH 2.0~12.0)에서 안정하였으나 6$0^{\circ}C$에서 처리하면 산에서는 안정하였으나 Alkali에서는 불안정하였다.

One strain of Streptomyces sp. (AS-707) isolated from soil was found to produce a biologically active substance that showed a strong inhibitory activity against proteolytic enzymes viz. trypsin, papain, $\alpha$-chymotrypsin, Azotobacter protease, and Bacillus pretense. The substance was separated from culture filtrate by ion exchange column chromatography using Amberlite IRC-50 and CM-cellulose column chromatography. It was found that the recovery yield was 26% as activity basis. The substance was stable in wide pH range from 2.0 to 12.0 at 37$^{\circ}C$, but it was unstable in alkaline pH values at 6$0^{\circ}C$. The activity was thermostable to give 90% activity compared to the intact sample when it was treated at pH5.6 at 10$0^{\circ}C$ for 2 hours.