Kim Min-Jeong, Yuk Hyun-Gyun
Food Science and Technology Programme, Department of Chemistry, National University of Singapore, Singapore.
Food Science and Technology Programme, Department of Chemistry, National University of Singapore, Singapore
Appl Environ Microbiol. 2017 Feb 15;83(5). doi: 10.1128/AEM.02582-16. Print 2017 Mar 1.
The aim of this study was to elucidate the antibacterial mechanism of 405 ± 5-nm light-emitting diode (LED) illumination against at 4°C in phosphate-buffered saline (PBS) by determining endogenous coproporphyrin content, DNA oxidation, damage to membrane function, and morphological change. Gene expression levels, including of , , , , and , were also examined to understand the response of to LED illumination. The results showed that strains responded differently to LED illumination, revealing that serovar Enteritidis (ATCC 13076) and subsp. serovar Saintpaul (ATCC 9712) were more susceptible and resistant, respectively, than the 16 other strains tested. There was no difference in the amounts of endogenous coproporphyrin in the two strains. Compared with that in nonilluminated cells, the DNA oxidation levels in illuminated cells increased. In illuminated cells, we observed a loss of efflux pump activity, damage to the glucose uptake system, and changes in membrane potential and integrity. Transmission electron microscopy revealed a disorganization of chromosomes and ribosomes due to LED illumination. The levels of the five genes measured in the nonilluminated and illuminated Saintpaul cells were upregulated in PBS at a set temperature of 4°C, indicating that increased gene expression levels might be due to a temperature shift and nutrient deficiency rather than to LED illumination. In contrast, only in Enteritidis cells was upregulated. Thus, different sensitivities of the two strains to LED illumination were attributed to differences in gene regulation. Bacterial inactivation using visible light has recently received attention as a safe and environmentally friendly technology, in contrast with UV light, which has detrimental effects on human health and the environment. This study was designed to understand how 405 ± 5-nm light-emitting diode (LED) illumination kills strains at refrigeration temperature. The data clearly demonstrated that the effectiveness of LED illumination on strains depended highly on the serotype and strain. Our findings also revealed that its antibacterial mechanism was mainly attributed to DNA oxidation and a loss of membrane functions rather than membrane lipid peroxidation, which has been proposed by other researchers who studied the antibacterial effect of LED illumination by adding exogenous photosensitizers, such as chlorophyllin and hypericin. Therefore, this study suggests that the detailed antibacterial mechanisms of 405-nm LED illumination without additional photosensitizers may differ from that by exogenous photosensitizers. Furthermore, a change in stress-related gene regulation may alter the susceptibility of cells to LED illumination at refrigeration temperature. Thus, our study provides new insights into the antibacterial mechanism of 405 ± 5-nm LED illumination on cells.
本研究的目的是通过测定内源性粪卟啉含量、DNA氧化、膜功能损伤和形态变化,阐明405±5纳米发光二极管(LED)照射在4℃磷酸盐缓冲盐水(PBS)中对[细菌名称未给出]的抗菌机制。还检测了包括[基因名称未给出]等基因的表达水平,以了解[细菌名称未给出]对LED照射的反应。结果表明,[细菌名称未给出]菌株对LED照射的反应不同,显示肠炎血清型[细菌名称未给出](ATCC 13076)和[细菌名称未给出]亚种圣保罗血清型[细菌名称未给出](ATCC 9712)分别比其他16株测试菌株更敏感和更具抗性。两株菌株内源性粪卟啉的含量没有差异。与未照射细胞相比,照射细胞中的DNA氧化水平增加。在照射细胞中,我们观察到外排泵活性丧失、葡萄糖摄取系统受损以及膜电位和完整性的变化。透射电子显微镜显示由于LED照射导致染色体和核糖体紊乱。在4℃设定温度下,在未照射和照射的圣保罗[细菌名称未给出]细胞中测得的五个基因的水平在PBS中上调,表明基因表达水平升高可能是由于温度变化和营养缺乏,而不是LED照射。相比之下,肠炎[细菌名称未给出]细胞中只有[基因名称未给出]上调。因此,两株菌株对LED照射的不同敏感性归因于基因调控的差异。与对人类健康和环境有有害影响的紫外线相比,利用可见光进行细菌灭活作为一种安全且环保的技术最近受到了关注。本研究旨在了解405±5纳米发光二极管(LED)照射如何在冷藏温度下杀死[细菌名称未给出]菌株。数据清楚地表明,LED照射对[细菌名称未给出]菌株的有效性高度依赖于血清型和菌株。我们的研究结果还表明,其抗菌机制主要归因于DNA氧化和膜功能丧失,而不是膜脂质过氧化,其他研究人员通过添加外源性光敏剂(如叶绿酸和金丝桃素)来研究LED照射的抗菌作用时曾提出膜脂质过氧化。因此,本研究表明,无额外光敏剂的405纳米LED照射的详细抗菌机制可能与外源性光敏剂的不同。此外,应激相关基因调控的变化可能会改变[细菌名称未给出]细胞在冷藏温度下对LED照射的敏感性。因此,我们的研究为405±5纳米LED照射对[细菌名称未给出]细胞的抗菌机制提供了新的见解。