Carré Gaëlle, Hamon Erwann, Ennahar Saïd, Estner Maxime, Lett Marie-Claire, Horvatovich Peter, Gies Jean-Pierre, Keller Valérie, Keller Nicolas, Andre Philippe
Laboratoire de Biophotonique et Pharmacologie, UMR 7213 CNRS and Strasbourg University, Illkirch, France.
Appl Environ Microbiol. 2014 Apr;80(8):2573-81. doi: 10.1128/AEM.03995-13. Epub 2014 Feb 14.
This study investigates the mechanisms of UV-A (315 to 400 nm) photocatalysis with titanium dioxide (TiO2) applied to the degradation of Escherichia coli and their effects on two key cellular components: lipids and proteins. The impact of TiO2 photocatalysis on E. coli survival was monitored by counting on agar plate and by assessing lipid peroxidation and performing proteomic analysis. We observed through malondialdehyde quantification that lipid peroxidation occurred during the photocatalytic process, and the addition of superoxide dismutase, which acts as a scavenger of the superoxide anion radical (O2·(-)), inhibited this effect by half, showing us that O2·(-) radicals participate in the photocatalytic antimicrobial effect. Qualitative analysis using two-dimensional electrophoresis allowed selection of proteins for which spot modifications were observed during the applied treatments. Two-dimensional electrophoresis highlighted that among the selected protein spots, 7 and 19 spots had already disappeared in the dark in the presence of 0.1 g/liter and 0.4 g/liter TiO2, respectively, which is accounted for by the cytotoxic effect of TiO2. Exposure to 30 min of UV-A radiation in the presence of 0.1 g/liter and 0.4 g/liter TiO2 increased the numbers of missing spots to 14 and 22, respectively. The proteins affected by photocatalytic oxidation were strongly heterogeneous in terms of location and functional category. We identified several porins, proteins implicated in stress response, in transport, and in bacterial metabolism. This study reveals the simultaneous effects of O2·(-) on lipid peroxidation and on the proteome during photocatalytic treatment and therefore contributes to a better understanding of molecular mechanisms in antibacterial photocatalytic treatment.
本研究调查了应用二氧化钛(TiO₂)进行紫外线A(315至400纳米)光催化降解大肠杆菌的机制及其对两个关键细胞成分:脂质和蛋白质的影响。通过在琼脂平板上计数、评估脂质过氧化和进行蛋白质组分析来监测TiO₂光催化对大肠杆菌存活的影响。我们通过丙二醛定量观察到在光催化过程中发生了脂质过氧化,而作为超氧阴离子自由基(O₂·(-))清除剂的超氧化物歧化酶的添加将这种效应抑制了一半,这表明O₂·(-)自由基参与了光催化抗菌作用。使用二维电泳的定性分析允许选择在应用处理过程中观察到斑点修饰的蛋白质。二维电泳突出显示,在所选蛋白质斑点中,分别在存在0.1克/升和0.4克/升TiO₂的黑暗环境中,有7个和19个斑点已经消失,这是由TiO₂的细胞毒性作用导致的。在存在0.1克/升和0.4克/升TiO₂的情况下暴露于30分钟的紫外线A辐射,分别使缺失斑点的数量增加到14个和22个。受光催化氧化影响的蛋白质在位置和功能类别方面具有很强的异质性。我们鉴定出了几种孔蛋白、与应激反应、运输和细菌代谢有关的蛋白质。本研究揭示了在光催化处理过程中O₂·(-)对脂质过氧化和蛋白质组的同时作用,因此有助于更好地理解抗菌光催化处理中的分子机制。