Mechanism of Bacillus subtilis spore inactivation by and resistance to supercritical CO2 plus peracetic acid.
Mechanism of Bacillus subtilis spore inactivation by and resistance to supercritical CO2 plus peracetic acid.
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DOI：
10.1111/jam.12995
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发表时间：
2016-01
影响因子：
4
通讯作者：
Setlow P
中科院分区：
文献类型：
--
作者：
Setlow B;Korza G;Blatt KM;Fey JP;Setlow P
确定超临界CO2（scCO 2）加过氧乙酸（PAA）如何灭活枯草芽孢杆菌孢子，孢子对scCO 2-PAA的抗性的重要因素，以及scCO 2-PAA灭活的孢子是否真正死亡。野生型B的孢子。枯草芽孢杆菌和缺乏孢子保护蛋白的同基因突变体在35°C下用液体或干燥的scCO 2-PAA处理。野生型湿孢子（水悬浮液）比干孢子更敏感。检查经处理的孢子的活力（并且是真正死亡的）、吡啶二羧酸（DPA）、突变、对核酸染色剂的渗透性、在不同条件下的萌发、能量代谢和生长。ScCO 2-PAA灭活的孢子保留DPA，存活者没有明显的DNA损伤。然而，DPA在通常无害的温度（85°C）下从灭活的孢子中释放，并且来自处理的孢子的菌落形成是盐敏感的。灭活的孢子萌发但不长出，并且这些萌发的孢子具有改变的质膜透性和缺陷的能量代谢。湿或干的涂层缺陷的孢子增加scCO 2-PAA的敏感性，和干孢子，但不是湿孢子缺乏DNA保护蛋白的scCO 2-PAA更敏感。这些发现表明scCO 2-PAA通过破坏孢子的内膜来灭活孢子。孢子衣为湿孢子和干孢子提供scCO 2-PAA抗性。DNA保护蛋白仅对干孢子提供scCO 2-PAA抗性。这些结果提供了关于scCO 2-PAA（一种在灭菌应用中使用越来越多的试剂）的孢子灭活和抗性机制的信息。
Determine how supercritical CO2 (scCO2) plus peracetic acid (PAA) inactivates Bacillus subtilis spores, factors important in spore resistance to scCO2-PAA, and if spores inactivated by scCO2-PAA are truly dead. Spores of wild-type B. subtilis and isogenic mutants lacking spore protective proteins were treated with scCO2-PAA in liquid or dry at 35°C. Wild-type wet spores (aqueous suspension) were more susceptible than dry spores. Treated spores were examined for viability (and were truly dead), dipicolinic acid (DPA), mutations, permeability to nucleic acid stains, germination under different conditions, energy metabolism and outgrowth. ScCO2-PAA-inactivated spores retained DPA, and survivors had no notable DNA damage. However, DPA was released from inactivated spores at a normally innocuous temperature (85°C), and colony formation from treated spores was salt sensitive. The inactivated spores germinated but did not outgrow, and these germinated spores had altered plasma membrane permeability and defective energy metabolism. Wet or dry coat-defective spores had increased scCO2-PAA sensitivity, and dry spores but not wet spores lacking DNA protective proteins were more scCO2-PAA sensitive. These findings suggest that scCO2-PAA inactivates spores by damaging spores’ inner membrane. The spore coat provided scCO2-PAA resistance for both wet and dry spores. DNA protective proteins provided scCO2-PAA resistance only for dry spores. These results provide information on mechanisms of spore inactivation of and resistance to scCO2-PAA, an agent with increasing use in sterilization applications.