산내성 섬유소 분해 박테리아 분리 및 효소학적 특성규명
Isolation and Biochemical Characterization of Acid Tolerance Bacteria from Camellia sinensis Planting Soil
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본 연구는 차나무식재지 토양에서 LB-CMC 한천배지를 이용하여 목질계 바이오매스 분해능이 우수한 미생물을 분리하고자 실시하였다. 그결과 목질계 바이오매스분해에 우수한 활성을 보이는 3종의 박테리아를 분리하였다. 16S rRNA 유전자를 이용하여 3종의 박테리아를 동정한결과 모두 Bacillus속에 속하였다. CMC zymogram 분석결과 Bacullus 3종 모두 약 44kDa의 셀룰레이즈가 존재하였다. Bacillus sp. CS1의최적생장 온도는 37℃였으며, CMCase의 최대활성은 배양 36시간 후였고, xylanase는 배양 후 12시간에 최대활성을 보였다. CMCase와 xylanase의최적 pH는 각각 5.0이었다. CMCase의 열안정성은 높았지만, xylanse는 열에 불안정한 것을 보였다. Bacillus sp. CS2의 최적 생장온도는 37℃였으며, CMCase와 xylanaase의 활성은 배양 후 36시간이 가장 높았다. CMCase의 최적 온도와 pH는 각각 37℃와 pH 4.0이었지만, xylanaase의최적 온도와 pH는 50℃와 pH 5.0이었다. CMCase와 xylanase는 비교적 열에 안정적이었다. Bacillus sp. CS3 최적 생장 온도는 37℃였으며,CMCase와 xylanase의 활성은 배양 후 36시간이 가장 높았다. CMCase 및 xylanase에 대한 최적의 온도와 pH는 37℃와 4.0이었다. CMCase의열안정성을 보였지만, xylanase는 열에 불안정한 것을 보였다.
Abstract
We attempted to isolate microorganisms with excellent ability to degrade lignocellulosic biomass using LB-CMC agar plate from Camelliasinensis planting soil. As a result, three bacteria were isolated that showed excellent activity in lignocellulosic biomass degradation. The16S rRNA gene was used to identify three newly isolated bacteria and three bacteria were genus Bacillus. Bacteria were named as Bacillussp. CS1, Bacillus sp. CS2, and Bacillus sp. CS3. In the CMC zymogram analysis of three bacteria, it showed that one active band ofabout 44kDa is present each bacterium. The optimal growth of Bacillus sp. CS1 was 37℃. The maximal activity for CMCase showedafter 36hr of cultivation and the maximal activity for xylanase was 12hr of cultivation. The optimal temperature and pH for CMCasewere 37℃ and 5.0, respectively, but for xylanase were 50℃ and 5.0, respectively. CMCase from Bacillus sp. CS1 showed to be thermalstable, but xylanase was thermally unstable. The optimal growth temperature of Bacillus sp. CS2 was 37℃. The activity of both CMCaseand xylanase reached the highest at 36hr culture. The optimal temperature and pH for CMCase from Bacillus sp. CS2 were 37℃ andpH 4.0, respectively, but the optimal temperature and pH for xylanase were 50℃ and pH 5.0. Both thermal stability of CMCase andxylanase were relatively stable. The optimal growth temperature of Bacillus sp. CS3 was 37℃. Both activity of CMCase and xylanasereached the highest at 36hr culture. The optimal temperature and pH for CMCase and xylanase from Bacillus sp. CS3 were 37℃ and4.0, respectively. CMCase from Bacillus sp. CS3 showed to be thermal stable, but xylanase exhibited thermally unstable.
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