RESUMEN
Blue light (BL) exerts an antimicrobial effect on pathogenic bacteria. It has been hypothesized that its bactericidal activity depends upon the generation of reactive oxygen species (such as anion superoxides) and the resultant cellular damage. However, some aspects of this hypothesis needed to be tested and investigated. Thus, the work conducted herein examined the molecular impact of BL treatment on Cronobacter sakazakii, an emerging foodborne pathogen. The results showed that BL exhibited an efficient bactericidal effect against C. sakazakii. Under a sublethal BL dose, both intracellular anion superoxides and malondialdehyde (a marker of oxidative stress) contents were increased gradually. Moreover, permeability of the outer membrane was increased by approximately 50%, indicating membrane damage. Further investigation revealed alterations to cellular fatty acid profiles, with a decrease and disappearance of unsaturated fatty acids, including C18:2, C16:1, and C18:1. These data indicate that bacterial lipids, especially unsaturated fatty acids, are important molecular targets of BL photo-oxidation. The transcriptional response of bacteria to BL was also studied, and it was found that three genes were upregulated, including genes encoding antioxidants. The current study contributes towards an improved understanding of the bactericidal mechanisms of BL and highlights the importance of lipid and membrane damage.
Asunto(s)
Cronobacter sakazakii/efectos de la radiación , Ácidos Grasos/efectos de la radiación , Luz , Estrés Oxidativo/efectos de la radiación , Membrana Externa Bacteriana/metabolismo , Membrana Externa Bacteriana/efectos de la radiación , Cronobacter sakazakii/genética , Cronobacter sakazakii/metabolismo , Ácidos Grasos/química , Genes Bacterianos/genética , Viabilidad Microbiana/efectos de la radiación , Especies Reactivas de Oxígeno/metabolismo , Regulación hacia Arriba/efectos de la radiaciónRESUMEN
IMPORTANCE: New approaches are needed to discover novel antimicrobials, particularly antibiotics that target the Gram-negative outer membrane. By exploiting bacterial sensing and responses to outer membrane (OM) damage, we used a biosensor approach consisting of polymyxin resistance gene transcriptional reporters to screen natural products and a small drug library for biosensor activity that indicates damage to the OM. The diverse antimicrobial compounds that cause induction of the polymyxin resistance genes, which correlates with outer membrane damage, suggest that these LPS and surface modifications also function in short-term repair to sublethal exposure and are required against broad membrane stress conditions.