Mizuno Akira
Toyohashi University of Technology, 1-1 Hibarigaoka, Tenpaku-cho, Toyohashi, Aichi 441-8580, Japan.
J Clin Biochem Nutr. 2017 Jan;60(1):12-24. doi: 10.3164/jcbn.16-64. Epub 2016 Dec 17.
Mechanism of inactivation of bio-particles exposed to non-thermal plasma (NTP), namely, dielectric barrier discharge (DBD), and plasma jet (PJ), has been studied using , spore, and bacteriophages. States of different biological components were monitored during the course of inactivation. Analysis of green fluorescent protein, GFP, introduced into . or spore cells proved that radicals generated by NTP penetrate into microbes, destroying the cell membrane and finally damage the genes. We have evaluated the damage of the bacteriophages. Bacteriophage λ having double stranded DNA was exposed to DBD, then DNA was purified and subjected to DNA packaging reactions. The re-packaged phages consist of the DNA from discharged phages and brand-new coat proteins were proved to be active, indicating that the damage of coat proteins is responsible for inactivation. M13 phages having single stranded DNA were also examined with the same manner. In this case, damage to the DNA was as severe as that of the coat proteins. For practical applications, DBD showed very intense sterilization ability for spore with the D-value of less than 10 s. This result indicates a possibility of application of NTP for quick sterilization.
利用芽孢杆菌孢子和噬菌体研究了暴露于非热等离子体(NTP)(即介质阻挡放电(DBD)和等离子体射流(PJ))下生物颗粒的失活机制。在失活过程中监测了不同生物成分的状态。对导入芽孢杆菌孢子细胞或枯草芽孢杆菌孢子细胞中的绿色荧光蛋白(GFP)的分析证明,NTP产生的自由基穿透微生物,破坏细胞膜并最终损害基因。我们评估了噬菌体的损伤情况。将具有双链DNA的噬菌体λ暴露于DBD,然后纯化DNA并进行DNA包装反应。重新包装的噬菌体由放电噬菌体的DNA和全新的外壳蛋白组成,被证明具有活性,这表明外壳蛋白的损伤是失活的原因。对具有单链DNA的M13噬菌体也采用相同的方式进行了检测。在这种情况下,DNA的损伤与外壳蛋白的损伤一样严重。对于实际应用,DBD对枯草芽孢杆菌孢子显示出非常强的杀菌能力,D值小于10秒。这一结果表明NTP有可能用于快速杀菌。