Label-Free Proteomic Approach to Study the Non-lethal Effects of Silver Nanoparticles on a Gut Bacterium (original) (raw)
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Biochemical and Biophysical Research Communications, 2012
Despite extensive use of silver nanoparticles for antimicrobial applications, cellular mechanisms underlying microbial response to silver nanoparticles remain to be further elucidated at the systems level. Here, we report systems-level response of Escherichia coli to silver nanoparticles using transcriptomebased biochemical and phenotype assays. Notably, we provided the evidence that anaerobic respiration is induced upon exposure to silver nanoparticles. Further we showed that anaerobic respiration-related regulators and enzymes play an important role in E. coli resistance to silver nanoparticles. In particular, our results suggest that arcA is essential for resistance against silver NPs and the deletion of fnr, fdnH and narH significantly increases the resistance. We envision that this study offers novel insights into modes of antimicrobial action of silver nanoparticles, and cellular mechanisms contributing to the development of microbial resistance to silver nanoparticles.
Investigating Fate of Silver Nanoparticles in Wastewater Biofilms
2017
As industrial advances make everyday life easier for human kind, the processes by which we need to maintain sanitary conditions for both water and wastewater treatment will become increasingly complex. Innovations in food packaging and textile design incorporate engineered nanoparticles (ENPs) to increase antimicrobial properties of clothing, maintain product color, and keep food in packaging from spoilage. For most products, ENPs released will enter the sanitary sewer system, and ultimately wastewater treatment plants. Biofilms grow universally on surfaces where a protective layer of extracellular polymeric substances (EPS) shields attached cells from stressors. In wastewater treatment, complex biofilms are utilized as a biological process for nutrient removal. Along with manufacturing innovations, the technology to study wastewater processes also continues to advance. Understanding complex biological communities requires detailed expertise in metagenomics for identifying bacteria ...
The international journal of biochemistry & cell biology, 2018
The oxidative stress generation in bacteria by the presence of antibiotics (in this case silver nanoparticles (AgNPs)) is already widely known. Previously, we demonstrated that AgNPs generate oxidative stress in Staphylococcus aureus and Escherichia coli mediated by the increase of reactive oxygen species (ROS). In this work we are demonstrating the consequences of the oxidative stress by the presence of AgNPs; these bacterial strains increased the levels of oxidized proteins and lipids. In addition, it was possible to determine which reactive oxygen species are mainly responsible for the oxidative damage to macromolecules. Also, we found that the bacterial DNA was fragmented and the membrane potential was modified. This increase in the levels of ROS found in both, S. aureus and E. coli, was associated with the oxidation of different types of important macromolecules for the normal functioning of cell, so the oxidative stress would be one of the mechanisms by which the AgNPs would e...
Biointerphases, 2016
The present work is aimed at comparing the effects of sublethal concentrations of silver nanoparticles (AgNPs) on the growth kinetic, adhesion ability, oxidative stress, and phenotypic changes of model bacteria (Escherichia coli and Bacillus subtilis) under both aerobic and anaerobic conditions. Growth kinetic tests conducted in 96-well microtiter plates revealed that sublethal concentrations of AgNPs do not affect E. coli growth, whereas 1 μg/ml AgNPs increased B. subtilis growth rate under aerobic conditions. At the same concentration, AgNPs promoted B. subtilis adhesion, while it discouraged E. coli attachment to the surface in the presence of oxygen. As determined by 2,7-dichlorofluorescein-diacetate assays, AgNPs increased the formation of intracellular reactive oxygen species, but not at the highest concentrations, suggesting the activation of scavenging systems. Finally, motility assays revealed that 0.01 and 1 μg/ml AgNPs, respectively, promoted surface movement in E. coli a...
Environmental Pollution, 2019
RUNNING TITLE Impact of silver nanoparticles on gut microbiota HIGHLIGHTS • Interactions among AgNPs, the intestinal microbiota, and a probiotic were tested • AgNPs did not affect the core fecal microflora and its metabolic and toxic profiles • Functional differences in few critical pathways occurred in AgNPs-treated samples • The probiotic had a chemopreventive role on fecal microflora against AgNPs CAPSULE This work will contribute to better understand the effects of non-lethal concentrations of AgNPs on the gut microbiota, and their interactions with probiotic administrations.
Environmental Science & Technology, 2010
Here we describe results from a proteomic study of protein-nanoparticle interactions to further the understanding of the ecotoxicological impact of silver nanoparticles (AgNPs) in the environment. We identified a number of proteins from Escherichia coli that bind specifically to bare or carbonatecoated AgNPs. Of these proteins, tryptophanase (TNase) was observed to have an especially high affinity for both surface modifications despite its low abundance in E. coli. Purified TNase loses enzymatic activity upon associating with AgNPs, suggesting that the active site may be in the vicinity of the binding site(s). TNase fragments with high affinities for both types of AgNPs were identified using matrix-assisted laser desorption/ ionization time-of-flight (MALDI-TOF) mass spectrometry. Differences in peptide abundance/presence in mass spectra for the two types of AgNPs suggest preferential binding of some protein fragments based on surface coating. One highbinding protein fragment contained a residue (Arg103) that is part of the active site. Ag adducts were identified for some fragments and found to be characteristic of strong binding to AgNPs rather than association of the fragments with ionic silver. These results suggest a probable mechanism for adhesion of proteins to the most commonly used commercial nanoparticles and highlight the potential effect of nanoparticle surface coating on bioavailability.
Nanotoxicology
A c c e p t e d M a n u s c r i p t Time-resolved toxicity study reveals the dynamic interactions between uncoated silver nanoparticles and bacteria It is still unclear whether the toxicity of silver nanoparticles (AgNPs) can be attributed solely to the release of Ag + or whether dissolved and nanoparticulate Ag act in parallel; this is due to the difficulty in distinguishing Ag +-from AgNP-effects. Also, AgNPs undergo changes during toxicity tests. This is the first study to investigate the influence of AgNP dissolution over time on viable counts at high time resolution and low cell density, avoiding the apparently reduced toxicity at higher cell densities identified in our study. Uncapped AgNPs were synthesized to avoid any interference from surface coatings. The transformations of AgNPs during storage were reduced. Lowering the concentration of AgNPs reduced their aggregation in Davis minimal medium (DMM). Also, AgNPs dissolved more slowly in DMM than in water. The minimum inhibitory concentrations (MICs) of Ag + and AgNPs increased with cell density according to a power law, suggesting that binding to cells decreased effective concentrations. However, AgNPs acted as a reservoir of Ag, releasing new Ag + to maintain the Ag stress. The toxicity of AgNPs was dominated by dissolved Ag. Combining controlled conditions, high time-resolution and low cell density, we could demonstrate different roles of ionic and nano Ag in bacterial death caused by AgNPs.
Silver nanoparticles restrict microbial growth by promoting oxidative stress and DNA damage
EXCLI Journal, 2020
Bacterial infections remain a serious health issue; hence there is a need for continuous search for improved antimicrobials. In addition, it is important to understand the antibacterial mechanism of prospective antimicrobials to fully harness their benefits. In this study, the antimicrobial action of silver nanoparticles was investigated. The antimicrobial potential of silver nanoparticles against different strains of bacteria was evaluated after which Escherichia coli and Staphylococcus aureus were selected as model for gram-negative and gram-positive bacteria respectively. Additionally, to determine mechanism of action, some biochemical assays including determination of kynurenine level, DNA fragmentation, lipid peroxidation and antioxidant status were carried out. Results showed that silver nanoparticles caused DNA damage and induced oxidative stress as reflected in elevated nitric oxide production and lipid peroxidation level. In contrast silver nanoparticles increased the antio...
Silver nanoparticles (AgNP) are widely used in consumer products, mostly due to their excellent antimicrobial properties. One of the well-established antibacterial mechanisms of AgNP is their efficient contact with bacteria and dissolution on cell membranes. To our knowledge, the primary mechanism of cell wall damage and the event(s) initiating bactericidal action of AgNP are not yet elucidated. In this study we used a combination of different assays to reveal the effect of AgNP on i) bacterial envelope in general, ii) outer membrane (OM) and iii) on plasma membrane (PM). We showed that bacterial PM was the main target of AgNP in Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa. AgNP depolarized bacterial PM, induced the leakage of the intracellular K + , inhibited respiration and caused the depletion of the intracellular ATP. In contrast, AgNP had no significant effect on the bacterial OM. Most of the adverse effects on bacterial envelope and PM occurred within th...