초록
고온성 Clostridium thermohydrosulfioicum의 autolysis 및 autoplast 형성에 대하여 조사하였다. 초기대수증식기 상태의 세포가 $K^{+}$ $Na^{+}$등의 1가이온을 함유한 buffer나 Tris-HCl buffer 속에서 autolysis 현상을 나타내었다. 이 현상은 1가이온이나 cysteine-HCl, sorbitol, glycerol 등의 화학물질이 존재할 때 강하게 촉진되었으나 $Mg^{2+},Mn^{2+},Ca^{2+},Ni^{2+}$등의 2가 이온에 의해 다소 저해되었으며, 특히 $Fe^{2+},Cu^{2+}$ 등의 2가이온 및 citric acid에 의해서는 강하게 저해되었다. 또한 세포벽합성 저해제인 ampicillin을 함유한 배지에서 생육한 세포는 autolysis가 촉진되었으나 핵산 및 단백질합성 저해제의 경우 autolysis가 저해되었다. Autolysis가 유도되는 최적 pH는 7.5, 최적 온도는 $60^{\circ}C$였다. 한편, autolysis가 일어나는 도중, $Mg^{2+}, Mn^{2+}, Ca^{2+}, Ni^{2+}$등의 2가 이온에 의해 원형질막이 안정화 될 때 lysozyme의 첨가없이 autoplast가 형성되었다. 이 autoplast는 대수증식기 말기 세포에서 형성이 잘 되었으며, autoplast 형성의 최적 pH는 7.5, 최적 온도는 $37^{\circ}C$, 최적 $MgCl^{2}$ 농도는 20mM이었으며, 화학물질로는 0.3M glycerol이 효과적이었다.
Induction conditions for autolysis and autoplast formation of thermophilic Clostridium thermohydrosulfuricum were studied. The cells in the initial exponential growth phase were well autolysed in Tris-HCl buffer or inorganic buffers containing univalents, such as $K^{+}$ and $Na^{+}$ , and chemicals such as cysteine-HCl, sorbitol and glycerol. Meanwhile, autolysis induction was slightly inhibited by divalents, such as $Mg^{2+}, Mn^{2+}, Ca^{2+}, Ni^{2+}$, and strongly by divalents, such as $Fe^{2+}, Cu^{2+}$ and citric acid. The autolysis was stimulated when the cells were grown in the medium containing ampicillin that inhibites cell wall synthesis, meanwhile, it was slightly inhibited by nucleic acids and protein synthesis inhibitors. The optimal pH and temperature for the induction of autolysis were 7.5 and $60^{\circ}C$, respectively. On the other hand, the cells were autoplasted without lysozyme treatment during autolysis due to the stabilization of protoplasmic membrane in the presence of divalents such as $Mg^{2+}, Mn^{2+}, Ca^{2+}, Ni^{2+}$. Autoplast formation was mostly induced at $37^{\circ}C$ in 50mM Tris-HCl buffer (pH 7.5) containing 20 mM $MgCl^{2}$ and 0.3 M glycerol, and in the late exponential growth phase growing cell.