Mechanisms of killing of spores of Bacillus subtilis by iodine, glutaraldehyde and nitrous acid
Mechanisms of killing of spores of Bacillus subtilis by iodine, glutaraldehyde and nitrous acid
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DOI：
10.1046/j.1365-2672.2000.01114.x
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发表时间：
2000-08-01
影响因子：
4
通讯作者：
Setlow, P
中科院分区：
文献类型：
--
作者：
Tennen, R;Setlow, B;Setlow, P
用戊二醛或碘基消毒剂（聚维酮碘）处理枯草芽孢杆菌野生型孢子不会引起可检测到的诱变，并且缺乏主要 DNA 保护性 α/β 型小酸溶蛋白 (SASP) 的孢子（称为 α(-)β(-)）对这些试剂表现出相似的敏感性。 recA突变不会使野生型或α(-)β(-)孢子对聚维酮碘或戊二醛敏感，当处理的孢子萌发时，用这些试剂处理孢子也不会导致recA-lacZ融合体的显着表达。通过去除大量孢子外壳蛋白，孢子戊二醛敏感性显着增加，但这种处理对聚维酮碘敏感性没有影响。相比之下，野生型和α(-)β(-)孢子的亚硝酸处理引起显着的诱变，其中α(-)β(-)孢子对该试剂更加敏感。 recA突变进一步使野生型和α(-)β(-)孢子对亚硝酸敏感，并且当亚硝酸处理的孢子萌发时，recA-lacZ融合体显着表达。这些结果表明： (a) 亚硝酸至少部分通过 DNA 损伤杀死枯草芽孢杆菌孢子，而 α/β 型 SASP 可以防止这种 DNA 损伤； (b) 戊二醛或聚维酮碘杀死孢子不是由于 DNA 损伤； (c) 孢子衣可以保护孢子不被戊二醛杀死，但不能被聚维酮碘杀死。进一步的分析还表明，用亚硝酸处理的孢子仍能正常萌发，而用戊二醛或聚维酮碘处理的孢子却不能正常萌发。
Treatment of wild-type spores of Bacillus subtilis with glutaraldehyde or an iodine-based disinfectant (Betadine) did not cause detectable mutagenesis, and spores (termed alpha(-)beta(-)) lacking the major DNA-protective alpha/beta-type, small, acid-soluble proteins (SASP) exhibited similar sensitivity to these agents. A recA mutation did not sensitize wild-type or alpha(-)beta(-) spores to Betadine or glutaraldehyde, nor did spore treatment with these agents result in significant expression of a recA-lacZ fusion when the treated spores germinated. Spore glutaraldehyde sensitivity was increased dramatically by removal of much spore coat protein, but this treatment had no effect on Betadine sensitivity. In contrast, nitrous acid treatment of wild-type and alpha(-)beta(-) spores caused significant mutagenesis, with alpha(-)beta(-) spores being much more sensitive to this agent. A recA mutation further sensitized both wild-type and alpha(-)beta(-) spores to nitrous acid, and there was significant expression of a recA-lacZ fusion when nitrous acid-treated spores germinated. These results indicate that: (a) nitrous acid kills B. subtilis spores at least in part by DNA damage, and alpha/beta-type SASP protect against this DNA damage; (b) killing of spores by glutaraldehyde or Betadine is not due to DNA damage; and (c) the spore coat protects spores against killing by glutaraldehyde but not Betadine. Further analysis also demonstrated that spores treated with nitrous acid still germinated normally, while those treated with glutaraldehyde or Betadine did not.