热结合乙醇对芽孢的杀灭效果及内膜通透性的影响

Effect of Heat Combined with Ethanol on Spore Killing and Permeability of Inner Membrane

摘要:

研究了热结合乙醇对芽孢杀灭效果及内膜通透性的影响。采用平板计数法、荧光偏振法、分光光度法和流式细胞术对热结合乙醇处理后枯草杆菌芽孢的存活率、内膜流动性、OD₆₀₀值及内膜通透性进行了研究。结果显示，热结合乙醇能够杀灭芽孢，80 ℃结合体积分数75%乙醇处理后，芽孢存活浓度下降约1个对数值。60、80 ℃结合75%乙醇处理后，芽孢悬浮液的荧光偏振度下降了0.31，表明芽孢内膜流动性大幅增加。80 ℃结合75%乙醇处理后OD₆₀₀值下降程度最大，芽孢内膜通透性发生显著变化，流式细胞术结果显示，阳性区域变化93.69%。研究表明，热结合乙醇处理下，芽孢内膜通透性的改变是芽孢死亡的重要原因之一。

Abstract:

The effect of heat combined with ethanol on the spore killing efficacy and permeability of inner membrane was studied in this paper. The survival rate, inner membrane fluidity, OD₆₀₀ value and inner membrane permeability of Bacillus subtilis spores treated by heat combined with ethanol were studied by plate counting, fluorescence polarization, spectrophotometry and flow cytometry. The results showed that heat combined ethanol could kill the spores, and the survival concentration of spore decreased by about one logarithm at 80 ℃ combined with 75% ethanol. The fluorescence polarization degree of spore suspension decreased by 0.31 at 60 ℃ and 80 ℃ combined with 75% ethanol, indicating that the fluidity of spore inner membrane increased greatly. The OD₆₀₀ value decreased the most after 80 ℃ and 75% ethanol treatment, and the permeability of spore inner membrane changed significantly. The results of flow cytometry showed that the positive area changed 93.69%. The results showed that the change of endomembrane permeability was one of the important reasons of spore death.