새로운 방선균의 탐색과 신규 유용 물질의 탐색 및 개발에 관한 연구 = Research on the Screeing and Development of New Biochemical and New Actiinomycetes

국문 초록 (Abstract)

토양으로부터 690종의 방선균을 분리 그중 α-amylase inhibitor의 활성이 큰 Streptomyces를 분리하였다. α-amylase inhibitor의 최적 생산 조건은 2% soluble starch, 2% trypton, 0.05% ZnSO₄초기 배양액 pH=7.0, 배양시간 60시간, 배양온도 30℃였다. 세포벽 성분을 분석한 결과 LL-DAP형이고, 단당류 분석결과 Arabinose, Galactose, Xylose등이 존재하지 않았다 따라서 Bergey's mannual of determinative bacteriology, I.S.P strain Key, Shirling과 Gottlieb, Bergey's mannual of sysmatic bateriology에 의해 cell wall type Ⅰ에 해당되고 생리, 생화학적 특징 및 형태적 특징으로 볼 때 α-amylase inhibitor 생성균주는 Streptomyces 에 속하므로 Streptomyces sp. GG 330-2로 명명하고 Streptomyces albulus와 유사한 균주로 동정했다.
최적온도, pH 및 배지에서 배양한 여액을 FPLC로 정제하였으며 α-amylase inhibitor의 안전성은 열처리 및 pH 변화 등에서 활성이 그대로 유지되는 안정성을 보였다.

다국어 초록 (Multilingual Abstract)

Various methods for selective isolation are being used in the ecological study of Actinomycetes and the search for useful strains of these organisms. Because of the difficulties of selective isolation of Actinomycetes, we focus on new methods being improved at our laboratory, which include pretreatment of soil suspension with spore activating agents and germicides. These treatments greatly increased the counts of Actinomycetes cfu(colony forming unit) from soil, decreased bacterial counts on the isolation plate of HY agar. Our treatment(dry heat, wet heat, phenol, SDS and Yeast extract) and HY agar were useful for the highly selectable isolation of Micromonospora and Actinomycetes. 690 strains of soil microorganism were isolated from Korean soil and compost samples. α-amylase inhibitor producing micro-organisms were isolated from blue zone separated Actinomycetes Agar piece on the starch agar plate contained α-amylase. The morphological, physiochemical characteristics of each strains and their culture condition were evaluated at 30℃ with the best medium containing 2% soluble starch, 2% trypton and 0.05% ZnSO₄and the activity was obtained after 60 hours culture. The purification of inhibitors was analyzed in order to know stability of temperature and pH. At 100℃ for 60 min and the range of pH 2∼8 the activity of inhibitor were stabilized. Strains producing α-amylase inhibitors were identified to be morphologically similar to Streptomyces albulus, each of which were analyzed by means of SEM, cell wall and monosaccharides.