Sucrose용액중(溶液中)에서 유기(誘起)되는 Clostridium saccharoperbutylacetonicum의 자기용해현상(自己溶解現象)에 의(依)하여 형성(形成)된 Protoplast의 성상(性狀)에 관(關)한 연구(硏究)

Biological Properties of Protoplasts Produced by Sucrose-induced Autolysis of Clostridium saccharoperbutylacetonicum

혐기성세균(嫌氣性細菌)인 C. saccharoperbutylacetonicum이 동조(同調) 농도(濃度)의 sucrose용액중(溶液中)에서 타(他) 용균효소(溶菌酵素) 또는 항생물질(抗生物質)을 첨가(添加)함이 없이 세포벽(細胞壁)이 급격(急激)히 용해(溶解)되어 구상(球狀)의 균(菌)으로 변화(變化)하는 특이(特異)한 현상(現象)이 확인(確認)되었다. 형성(形成)된 구상(球狀)의 균(菌)은 세포벽(細胞壁)이 완전(完全)히 제거(除去)된 원형질체(原形質體)로서 삼투압(壓) shock에 약(弱)하고 bacteriophage흡착능(吸着能)을 상실하였으며 분열증식(分裂增殖)이 불가능(不可能)하였다. 그러나 생체고분자(生體高分子)의 생합성(生合成)이 가능(可能)하며 mitomycin C처리(處理)에 의(依)하여 bacteriocin을 생산(生産)하였으며 분리(分離)한 phage DNA의 감염(感染)에 의(依)하여 자손(子孫) phage를 생산(生産)하였다. 즉(卽) 동조농도(同調濃度)의 sucrose 용액중(溶液中)에서 유기(誘起)되는 용균(溶菌)에 의(依)하여 생성(生成)된 구상(球狀)의 균(菌)은 세포벽(細胞壁)을 가지지 아니한 원형질체(原形質體)로서 모세포(母細胞)의 균학적(菌學的) 특성(特性)을 계승한 상태(狀態)에 있었으며 전형적(典型的)인 protoplast로 판단(判斷)되었다.

The young cells of Clostridiunm saccaroperbutylacetonicum were rapidly autolysed by exposing them to the hypertonic solution of sucrose(0.3-0.6M) without any other supplement to decompose the rigid cell wall. The cells were converted into the spherical cells by lysis. The spherical cells had following properties: (1) they were absent in the cell wall and osmotically fragile. (2) they were stabilized in the existence of 0.4M sucrose and 5mM $MgSO_4$ (3) they were resistant against adsorption of phage particles. (4) they allowed infection of the isolated phage DNA and produced progeny phage particles. (5) they were able to biosynthesize their macromolecules for a few hours according to a balanced manner of biosynthesis. (6) they were able to produce the bacteriocin particles by mitomycin C treatment. (7) they were unable to multiply. These results were all in the level of typical properties of bacterial protoplasts. It was apparent that the spherical cells formed by lysis occcurring by treatment with hypertonic sucrose were protoplasts.