초록

남조류 분해세균으로 알려진 Moraxella sp. CK-1의 세포외 자가용균효소의 특성을 규명하였다. Moraxella sp. CK-1은 대수증기식 초기부터 세포외부로 자가용균효소를 분비하며, 이들 효소는 60-$70^{\circ}C$, pH 9.0의 최적 활성 조건을 나타내었다. 이들 효소는 $Na^{+}$, $K^{+}$, $Li^{+}$등의 이온에 의해 활성이 촉진되고, $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$ 및 $Mn^{2+}$ 등의 이온에 의해 활성이 억제되며, 특히, $Fe^{2+}$와 $Cu^{2+}$ 이온 존재하에서는 효소의 활성이 거의 나타나지 않았다. Micrococcusluteus 세포를 기질로 이용하여 renaturing, SDS-PAGE를 수행한 결과 Moraxella sp. CK-1은 분자량이 각각 30,32,38 그리고 41 kDa인 4가지 효소를 분비하며, 이중 32와 41 kDa의 효소는 생장 초기부터 사멸기까지 지속적으로 배지에 분포하는 반면, 38 kDa의 효소는 대수증식기 중기 이전, 30 kDa의 효소는 대수증식기 중기 이후에 배지에 분포하였다. M. luteus의 SDS-insoluble peptidoglycan에 효소추출물을 처리하였을 때 반응액의 유리 아미노기 농도가 증가하는 것은 이들 효소가 N-acetylmuramyl-L-ananine amidase 또는 endopeptidase라는 사실을 시사한다.

Abstract

We studied the characteristics of extracellular autolysins from Moraxella sp. CK-1 which has been known to lyse the cyanobacterial cell walls. This bacterium excreted autolysins from the early exponential growth phase. These enzymes showed optimal action condition of 60-$70^{\circ}C$ and pH 9.0. Whereas $Na^{+}$, $K^{+}$ and $Li^{+}$ ions exhibited positive effect on the enzyme activity, $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$ and $Mn^{2+}$ ions exhibited negative effect. Especially, $Fe^{2+}$ and $Cu^{2+}$ ions almost completely suppressed the activity. Four extracellular autolysins of 30, 32, 38 and 41 kDa were detected in renaturing SOS-PAGE gel containing 0.2% heat-killed Micrococcus luteus cells as substrate. Among these 4 autolysins, 2 enzymes of 32 and 41 kDa distributed in the culture medium throughout the experimental time, but the 38 kOa enzyme diminished and 30 kOa began to appear at mid-exponential growth phase. When SOS-insoluble peptidoglycan of M. luteus was treated with the autolysins of Moraxella sp. CK-l, the concentration of free amino groups in reaction mixture increased. This indicates that the autolysins are N-acetylmuramyl-L-alanine amidase or endopeptidase.

주제어

#Autolysin   #renaturing SDS-PAGE   #zymogram   #Moraxella sp  

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