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Find video protocols related to scientific articles indexed in Pubmed.
Glutathione reductase-mediated synthesis of tellurium-containing nanostructures exhibiting antibacterial properties.
Appl. Environ. Microbiol.
PUBLISHED: 09-05-2014
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Tellurium, a metalloid belonging to group 16 of the periodic table, displays very interesting physical and chemical properties and lately has attracted significant attention for its use in nanotechnology. In this context, the use of microorganisms for synthesizing nanostructures emerges as an eco-friendly and exciting approach compared to their chemical synthesis. To generate Te-containing nanostructures, bacteria enzymatically reduce tellurite to elemental tellurium. In this work, using a classic biochemical approach, we looked for a novel tellurite reductase from the Antarctic bacterium Pseudomonas sp. strain BNF22 and used it to generate tellurium-containing nanostructures. A new tellurite reductase was identified as glutathione reductase, which was subsequently overproduced in Escherichia coli. The characterization of this enzyme showed that it is an NADPH-dependent tellurite reductase, with optimum reducing activity at 30°C and pH 9.0. Finally, the enzyme was able to generate Te-containing nanostructures, about 68 nm in size, which exhibit interesting antibacterial properties against E. coli, with no apparent cytotoxicity against eukaryotic cells.
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Global transcriptomic analysis uncovers a switch to anaerobic metabolism in tellurite-exposed Escherichia coli.
Res. Microbiol.
PUBLISHED: 04-05-2014
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Tellurite (TeO3(2-)) is harmful for most microorganisms, especially Gram-negative bacteria. Even though tellurite toxicity involves a number of individual aspects, including oxidative stress, malfunctioning of metabolic enzymes and a drop in the reduced thiol pool, among others, the general mechanism of toxicity is rather complex and not completely understood to date. This work focused on DNA microarray analysis to evaluate the Escherichia coli global transcriptomic response when exposed to the toxicant. Confirming previous results, the induction of the oxidative stress response regulator soxS was observed. Upregulation of a number of genes involved in the global stress response, protein folding, redox processes and cell wall organization was also detected. In addition, downregulation of aerobic respiration-related genes suggested a metabolic switch to anaerobic respiration. The expression results were validated through oxygen consumption experiments, which corroborated that tellurite-exposed cells effectively consume oxygen at lower rates than untreated controls.
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Tellurite reduction by Escherichia coli NDH-II dehydrogenase results in superoxide production in membranes of toxicant-exposed cells.
Biometals
PUBLISHED: 01-31-2014
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Tellurite, the most soluble tellurium oxyanion, is extremely harmful for most microorganisms. Part of this toxicity is due to the generation of reactive oxygen species that in turn cause oxidative stress. However, the way in which tellurite interferes with cellular processes is not well understood to date. Looking for new cellular tellurite targets, we decided to evaluate the functioning of the electron transport chain in tellurite-exposed cells. In this communication we show that the E. coli ndh gene, encoding NDH-II dehydrogenase, is significantly induced in toxicant-exposed cells and that the enzyme displays tellurite-reducing activity that results in increased superoxide levels in vitro.
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? -Ketoglutarate Accumulation Is Not Dependent on Isocitrate Dehydrogenase Activity during Tellurite Detoxification in Escherichia coli.
Biomed Res Int
PUBLISHED: 07-25-2013
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Tellurite is toxic to most microorganisms because of its ability to generate oxidative stress. However, the way in which tellurite interferes with cellular processes is not fully understood to date. In this line, it was previously shown that tellurite-exposed cells displayed reduced activity of the ? -ketoglutarate dehydrogenase complex ( ? -KGDH), which resulted in ? -ketoglutarate ( ? -KG) accumulation. In this work, we assessed if ? -KG accumulation in tellurite-exposed E. coli could also result from increased isocitrate dehydrogenase (ICDH) and glutamate dehydrogenase (GDH) activities, both enzymes involved in ? -KG synthesis. Unexpectedly both activities were found to decrease in the presence of the toxicant, an observation that seems to result from the decreased transcription of icdA and gdhA genes (encoding ICDH and GDH, resp.). Accordingly, isocitrate levels were found to increase in tellurite-exposed E. coli. In the presence of the toxicant, cells lacking icdA or gdhA exhibited decreased reactive oxygen species (ROS) levels and higher tellurite sensitivity as compared to the wild type strain. Finally, a novel branch activity of ICDH as tellurite reductase is presented.
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Monounsaturated fatty acids are substrates for aldehyde generation in tellurite-exposed Escherichia coli.
Biomed Res Int
PUBLISHED: 04-30-2013
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Reactive oxygen species (ROS) damage macromolecules and cellular components in nearly all kinds of cells and often generate toxic intracellular byproducts. In this work, aldehyde generation derived from the Escherichia coli membrane oxidation as well as membrane fatty acid profiles, protein oxidation, and bacterial resistance to oxidative stress elicitors was evaluated. Studies included wild-type cells as well as cells exhibiting a modulated monounsaturated fatty acid (MUFA) ratio. The hydroxyaldehyde 4-hydroxy 2-nonenal was found to be most likely produced by E. coli, whose levels are dependent upon exposure to oxidative stress elicitors. Aldehyde amounts and markers of oxidative damage decreased upon exposure to E. coli containing low MUFA ratios, which was paralleled by a concomitant increase in resistance to ROS-generating compounds. MUFAs ratio, lipid peroxidation, and aldehyde generation were found to be directly related; that is, the lower the MUFAs ratio, the lower the peroxide and aldehyde generation levels. These results provide additional evidence about MUFAs being targets for membrane lipid oxidation and their relevance in aldehyde generation.
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DNA, Cell Wall and General Oxidative Damage Underlie the Tellurite/Cefotaxime Synergistic Effect in Escherichia coli.
PLoS ONE
PUBLISHED: 01-01-2013
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The constant emergence of antibiotic multi-resistant pathogens is a concern worldwide. An alternative for bacterial treatment using nM concentrations of tellurite was recently proposed to boost antibiotic-toxicity and a synergistic effect of tellurite/cefotaxime (CTX) was described. In this work, the molecular mechanism underlying this phenomenon is proposed. Global changes of the transcriptional profile of Escherichia coli exposed to tellurite/CTX were determined by DNA microarrays. Induction of a number of stress regulators (as SoxS), genes related to oxidative damage and membrane transporters was observed. Accordingly, increased tellurite adsorption/uptake and oxidative injuries to proteins and DNA were determined in cells exposed to the mixture of toxicants, suggesting that the tellurite-mediated CTX-potentiating effect is dependent, at least in part, on oxidative stress. Thus, the synergistic tellurite-mediated CTX-potentiating effect depends on increased tellurite uptake/adsorption which results in damage to proteins, DNA and probably other macromolecules. Our findings represent a contribution to the current knowledge of bacterial physiology under antibiotic stress and can be of great interest in the development of new antibiotic-potentiating strategies.
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Glucose-6-phosphate dehydrogenase protects Escherichia coli from tellurite-mediated oxidative stress.
PLoS ONE
PUBLISHED: 06-27-2011
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The tellurium oxyanion tellurite induces oxidative stress in most microorganisms. In Escherichia coli, tellurite exposure results in high levels of oxidized proteins and membrane lipid peroxides, inactivation of oxidation-sensitive enzymes and reduced glutathione content. In this work, we show that tellurite-exposed E. coli exhibits transcriptional activation of the zwf gene, encoding glucose 6-phosphate dehydrogenase (G6PDH), which in turn results in augmented synthesis of reduced nicotinamide adenine dinucleotide phosphate (NADPH). Increased zwf transcription under tellurite stress results mainly from reactive oxygen species (ROS) generation and not from a depletion of cellular glutathione. In addition, the observed increase of G6PDH activity was paralleled by accumulation of glucose-6-phosphate (G6P), suggesting a metabolic flux shift toward the pentose phosphate shunt. Upon zwf overexpression, bacterial cells also show increased levels of antioxidant molecules (NADPH, GSH), better-protected oxidation-sensitive enzymes and decreased amounts of oxidized proteins and membrane lipids. These results suggest that by increasing NADPH content, G6PDH plays an important role in E. coli survival under tellurite stress.
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The Escherichia coli BtuE protein functions as a resistance determinant against reactive oxygen species.
PLoS ONE
PUBLISHED: 01-10-2011
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This work shows that the recently described Escherichia coli BtuE peroxidase protects the bacterium against oxidative stress that is generated by tellurite and by other reactive oxygen species elicitors (ROS). Cells lacking btuE (?btuE) displayed higher sensitivity to K(2)TeO(3) and other oxidative stress-generating agents than did the isogenic, parental, wild-type strain. They also exhibited increased levels of cytoplasmic reactive oxygen species, oxidized proteins, thiobarbituric acid reactive substances, and lipoperoxides. E. coli ?btuE that was exposed to tellurite or H(2)O(2) did not show growth changes relative to wild type cells either in aerobic or anaerobic conditions. Nevertheless, the elimination of btuE from cells deficient in catalases/peroxidases (Hpx(-)) resulted in impaired growth and resistance to these toxicants only in aerobic conditions, suggesting that BtuE is involved in the defense against oxidative damage. Genetic complementation of E. coli ?btuE restored toxicant resistance to levels exhibited by the wild type strain. As expected, btuE overexpression resulted in decreased amounts of oxidative damage products as well as in lower transcriptional levels of the oxidative stress-induced genes ibpA, soxS and katG.
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Tellurite-induced carbonylation of the Escherichia coli pyruvate dehydrogenase multienzyme complex.
Arch. Microbiol.
PUBLISHED: 07-02-2010
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The soluble tellurium oxyanion, tellurite, is toxic for most organisms. At least in part, tellurite toxicity involves the generation of oxygen-reactive species which induce an oxidative stress status that damages different macromolecules with DNA, lipids and proteins as oxidation targets. The objective of this work was to determine the effects of tellurite exposure upon the Escherichia coli pyruvate dehydrogenase (PDH) complex. The complex displays two distinct enzymatic activities: pyruvate dehydrogenase that oxidatively decarboxylates pyruvate to acetylCoA and tellurite reductase, which reduces tellurite (Te(4+)) to elemental tellurium (Te(o)). PDH complex components (AceE, AceF and Lpd) become oxidized upon tellurite exposure as a consequence of increased carbonyl group formation. When the individual enzymatic activities from each component were analyzed, AceE and Lpd did not show significant changes after tellurite treatment. AceF activity (dihydrolipoil acetyltransferase) decreased ~30% when cells were exposed to the toxicant. Finally, pyruvate dehydrogenase activity decreased >80%, while no evident changes were observed in complexs tellurite reductase activity.
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The Escherichia coli btuE gene, encodes a glutathione peroxidase that is induced under oxidative stress conditions.
Biochem. Biophys. Res. Commun.
PUBLISHED: 06-30-2010
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Most aerobic organisms are exposed to oxidative stress. Looking for enzyme activities involved in the bacterial response to this kind of stress, we focused on the btuE-encoded Escherichia coli BtuE, an enzyme that shares homology with the glutathione peroxidase (GPX) family. This work deals with the purification and characterization of the btuE gene product. Purified BtuE decomposes in vitro hydrogen peroxide in a glutathione-dependent manner. BtuE also utilizes preferentially thioredoxin A to decompose hydrogen peroxide as well as cumene-, tert-butyl-, and linoleic acid hydroperoxides, confirming that its active site confers non-specific peroxidase activity. These data suggest that the enzyme may have one or more organic hydroperoxide as its physiological substrate. The btuE gene was induced when cells were exposed to oxidative stress elicitors that included potassium tellurite, menadione and hydrogen peroxide, among others, suggesting that BtuE could participate in the E. coli response to reactive oxygen species. To our knowledge, this is the first report describing a glutathione peroxidase in E. coli.
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Simple, fast, and sensitive method for quantification of tellurite in culture media.
Appl. Environ. Microbiol.
PUBLISHED: 06-04-2010
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A fast, simple, and reliable chemical method for tellurite quantification is described. The procedure is based on the NaBH(4)-mediated reduction of TeO(3)(2-) followed by the spectrophotometric determination of elemental tellurium in solution. The method is highly reproducible, is stable at different pH values, and exhibits linearity over a broad range of tellurite concentrations.
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Tellurite-induced oxidative stress leads to cell death of murine hepatocarcinoma cells.
Biometals
PUBLISHED: 02-19-2010
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Data regarding tellurium (Te) toxicity are scarce. Studies on its metabolism, performed mainly in bacteria, underline a major role of reactive oxygen species (ROS). We investigated whether tellurite undergoes redox cycling leading to ROS formation and cancer cell death. The murine hepatocarcinoma Transplantable Liver Tumor (TLT) cells were challenged with tellurite either in the presence or in the absence of different compounds as N-acetylcysteine (NAC), 3-methyladenine, BAPTA-AM, and catalase. NAC inhibition of tellurite-mediated toxicity suggested a major role of oxidative stress. Tellurite also decreased both glutathione (GSH) and ATP content by 57 and 80%, respectively. In the presence of NAC however, the levels of such markers were almost fully restored. Tellurite-mediated ROS generation was assessed both by using the fluorescent, oxidation-sensitive probe dichlorodihydrofluorescein diacetate (DCHF-DA) and electron spin resonance (ESR) spectroscopy to detect hydroxyl radical formation. Cell death occurs by a caspase-independent mechanism, as shown by the lack of caspase-3 activity and no cleavage of poly(ADP-ribose)polymerase (PARP). The presence of gamma-H2AX suggests tellurite-induced DNA strand breaking, NAC being unable to counteract it. Although the calcium chelator BAPTA-AM did show no effect, the rapid phosphorylation of eIF2alpha suggests that, in addition to oxidative stress, an endoplasmic reticulum (ER) stress may be involved in the mechanisms leading to cell death by tellurite.
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Biological interactions of selenocyanate: bioprocessing, detection and toxicity.
Environ Technol
PUBLISHED: 12-03-2009
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The selenocyanate anion, SeCN(-), has been reported in wastewater from refineries whose petroleum comes from Se-rich marine shales. A metalloid-resistant bacterium was exposed to aqueous solutions of SeCN(-) to examine the relative toxicity of SeCN(-), and the results were compared with the toxicity of selenate and selenite and another G16 metalloid oxyanion, tellurite. We also determined the volatile organo-selenium species produced by bacterial cultures amended with selenocyanate anion, and we investigated a solid phase preconcentration technique for collecting SeCN(-) from aqueous samples with different ionic strengths and subsequent detection using capillary electrophoresis. The relative toxicity of SeCN(-) is comparable to that of selenate and selenite using the metalloid-resistant bacterium LHVE as the test organism. Tellurite was more toxic at all concentrations examined than all three selenium-containing anions, SeO4(2-), SeO3(2-), SeCN(-). Live cultures of LHVE amended with 1 mM NaSeCN produced volatile organo-sulphides and organo-selenides that could be collected in headspace using a solid phase microextraction fibre. The bioprocessing, i.e. the reduction and methylation of SeCN(-), is similar to that of selenate and selenite by other metalloid-resistant bacteria. An aqueous 1.0 mM solution of SeCN(-) could be captured from solution on solid-phase extraction (SPE) cartridges using an aminopropyl-based stationary phase. Selenocyanate anions, slowly pumped into a wetted SPE cartridge, were trapped on the cartridges solid phase and were subsequently eluted, thereby providing an increase in concentration above that of the original SeCN(-)-containing solution. Preconcentration factors of 3.9 were achieved using a mixed sodium hydroxide/methanol elution solvent and by adding NaCl to aqueous SeCN(-) before loading on the SPE cartridge.
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Production of dimethyl triselenide and dimethyl diselenenyl sulfide in the headspace of metalloid-resistant Bacillus species grown in the presence of selenium oxyanions.
Anal. Biochem.
PUBLISHED: 06-16-2009
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A Bacillus species harvested from the environment is metalloid resistant and, when grown anaerobically in complex growth medium and amended with the selenium oxyanion selenate, selenite, or selenocyanate, produces volatile organoselenium compounds in bacterial culture headspace. Two novel compounds so far undetected in bacterial culture headspace, CH3Se2SCH3 and CH3SeSeSeCH3, are produced and can be detected using solid-phase microextraction and gas chromatography with either fluorine-induced chemiluminescence or mass spectrometric detection. Differences in the electron impact fragmentation pattern of the mixed sulfur/selenide compounds allow the tentative differentiation between the symmetric and asymmetric isomers in this bacteriums headspace in favor of the asymmetric CH3SeSeSCH3 isomer.
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Tellurite-mediated disabling of [4Fe-4S] clusters of Escherichia coli dehydratases.
Microbiology (Reading, Engl.)
PUBLISHED: 04-21-2009
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The tellurium oxyanion tellurite is toxic for most organisms and it seems to alter a number of intracellular targets. In this work the toxic effects of tellurite upon Escherichia coli [4Fe-4S] cluster-containing dehydratases was studied. Reactive oxygen species (ROS)-sensitive fumarase A (FumA) and aconitase B (AcnB) as well as ROS-resistant fumarase C (FumC) and aconitase A (AcnA) were assayed in cell-free extracts from tellurite-exposed cells in both the presence and absence of oxygen. While over 90 % of FumA and AcnB activities were lost in the presence of oxygen, no enzyme inactivation was observed in anaerobiosis. This result was not dependent upon protein biosynthesis, as determined using translation-arrested cells. Enzyme activity of purified FumA and AcnB was inhibited when exposed to an in vitro superoxide-generating, tellurite-reducing system (ITRS). No inhibitory effect was observed when tellurite was omitted from the ITRS. In vivo and in vitro reconstitution experiments with tellurite-damaged FumA and AcnB suggested that tellurite effects involve [Fe-S] cluster disabling. In fact, after exposing FumA to ITRS, released ferrous ion from the enzyme was demonstrated by spectroscopic analysis using the specific Fe(2+) chelator 2,2-bipyridyl. Subsequent spectroscopic paramagnetic resonance analysis of FumA exposed to ITRS showed the characteristic signal of an oxidatively inactivated [3Fe-4S](+) cluster. These results suggest that tellurite inactivates enzymes of this kind via a superoxide-dependent disabling of their [4Fe-4S] catalytic clusters.
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Tellurite: history, oxidative stress, and molecular mechanisms of resistance.
FEMS Microbiol. Rev.
PUBLISHED: 04-02-2009
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The perceived importance of tellurium (Te) in biological systems has lagged behind selenium (Se), its lighter sister in the Group 16 chalcogens, because of telluriums lower crustal abundance, lower oxyanion solubility and biospheric mobility and the fact that, unlike Se, Te has yet to be found to be an essential trace element. Te applications in electronics, optics, batteries and mining industries have expanded during the last few years, leading to an increase in environmental Te contamination, thus renewing biological interest in Te toxicity. This chalcogen is rarely found in the nontoxic, elemental state (Te(0)), but its soluble oxyanions, tellurite (TeO(3)(2-)) and tellurate (TeO(4)(2-)), are toxic for most forms of life even at very low concentrations. Although a number of Te resistance determinants (Tel) have been identified in plasmids or in the bacterial chromosome of different species of bacteria, the genetic and/or biochemical basis underlying bacterial TeO(3)(2-) toxicity is still poorly understood. This review traces the history of Te in its biological interactions, its enigmatic toxicity, importance in cellular oxidative stress, and interaction in cysteine metabolism.
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Expression of the yggE gene protects Escherichia coli from potassium tellurite-generated oxidative stress.
Arch. Microbiol.
PUBLISHED: 01-22-2009
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Potassium tellurite is highly toxic to most forms of life and specific bacterial tellurite defense mechanisms are not fully understood to date. Recent evidence suggests that tellurite would exert its toxic effects, at least in part, through the generation of superoxide anion that occurs concomitantly with intracellular tellurite (Te(4+)) reduction to elemental tellurium (Te(o)). In this work the putative antioxidant role of YggE from Escherichia coli, a highly conserved protein in several bacterial species and whose function is still a matter of speculation, was studied. When exposed to tellurite, E. coli lacking yggE exhibited increased activity of superoxide dismutase and fumarase C, augmented levels of reactive oxygen species and high concentration of carbonyl groups in proteins. Upon genetic complementation with the homologous yggE gene these values were restored to those observed in the parental, isogenic, wild type strain, suggesting a direct participation of YggE in E. coli tolerance to tellurite.
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Expression of Aeromonas caviae ST pyruvate dehydrogenase complex components mediate tellurite resistance in Escherichia coli.
Biochem. Biophys. Res. Commun.
PUBLISHED: 01-13-2009
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Potassium tellurite (K(2)TeO(3)) is harmful to most organisms and specific mechanisms explaining its toxicity are not well known to date. We previously reported that the lpdA gene product of the tellurite-resistant environmental isolate Aeromonas caviae ST is involved in the reduction of tellurite to elemental tellurium. In this work, we show that expression of A. caviae ST aceE, aceF, and lpdA genes, encoding pyruvate dehydrogenase, dihydrolipoamide transacetylase, and dihydrolipoamide dehydrogenase, respectively, results in tellurite resistance and decreased levels of tellurite-induced superoxide in Escherichia coli. In addition to oxidative damage resulting from tellurite exposure, a metabolic disorder would be simultaneously established in which the pyruvate dehydrogenase complex would represent an intracellular tellurite target. These results allow us to widen our vision regarding the molecular mechanisms involved in bacterial tellurite resistance by correlating tellurite toxicity and key enzymes of aerobic metabolism.
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Cloning, purification and characterization of Geobacillus stearothermophilus V uroporphyrinogen-III C-methyltransferase: evaluation of its role in resistance to potassium tellurite in Escherichia coli.
Res. Microbiol.
PUBLISHED: 01-03-2009
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The Geobacillus stearothermophilus V cobA gene encoding uroporphyrinogen-III C-methyltransferase (also referred to as SUMT) was cloned into Escherichia coli and the recombinant enzyme was overexpressed and purified to homogeneity. The enzyme binds S-adenosyl-L-methionine and catalyzes the production of III methyl uroporphyrinogen in vitro. E. coli cells expressing the G. stearothermophilus V cobA gene exhibited increased resistance to potassium tellurite and potassium tellurate. Site-directed mutagenesis of cobA abolished tellurite resistance of the mesophilic, heterologous host and SUMT activity in vitro. No methylated, volatile derivatives of tellurium were found in the headspace of tellurite-exposed cobA-expressing E. coli, suggesting that the role of SUMT methyltransferase in tellurite(ate) detoxification is not related to tellurium volatilization.
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Tellurite enters Escherichia coli mainly through the PitA phosphate transporter.
Microbiologyopen
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Several transporters suspected to be involved in tellurite uptake in Escherichia coli were analyzed. Results showed that the PitA phosphate transporter was related to tellurite uptake. Escherichia coli ?pitA was approximately four-fold more tolerant to tellurite, and cell viability remained almost unchanged during prolonged exposure to the toxicant as compared with wild type or ?pitB cells. Notably, reduced thiols (toxicant targets) as well as superoxide dismutase, catalase, and fumarase C activities did not change when exposing the ?pitA strain to tellurite, suggesting that tellurite-triggered oxidative damage is attenuated in the absence of PitA. After toxicant exposure, remaining extracellular tellurite was higher in E. coli ?pitA than in control cells. Whereas inductively coupled plasma atomic emission spectrometric studies confirmed that E. coli ?pitA accumulates ?50% less tellurite than the other strains under study, tellurite strongly inhibited (32)P(i) uptake suggesting that the PitA transporter is one of the main responsible for tellurite uptake in this bacterium.
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Enhanced glutathione content allows the in vivo synthesis of fluorescent CdTe nanoparticles by Escherichia coli.
PLoS ONE
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The vast application of fluorescent semiconductor nanoparticles (NPs) or quantum dots (QDs) has prompted the development of new, cheap and safer methods that allow generating QDs with improved biocompatibility. In this context, green or biological QDs production represents a still unexplored area. This work reports the intracellular CdTe QDs biosynthesis in bacteria. Escherichia coli overexpressing the gshA gene, involved in glutathione (GSH) biosynthesis, was used to produce CdTe QDs. Cells exhibited higher reduced thiols, GSH and Cd/Te contents that allow generating fluorescent intracellular NP-like structures when exposed to CdCl(2) and K(2)TeO(3). Fluorescence microscopy revealed that QDs-producing cells accumulate defined structures of various colors, suggesting the production of differently-sized NPs. Purified fluorescent NPs exhibited structural and spectroscopic properties characteristic of CdTe QDs, as size and absorption/emission spectra. Elemental analysis confirmed that biosynthesized QDs were formed by Cd and Te with Cd/Te ratios expected for CdTe QDs. Finally, fluorescent properties of QDs-producing cells, such as color and intensity, were improved by temperature control and the use of reducing buffers.
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Tellurite-exposed Escherichia coli exhibits increased intracellular ?-ketoglutarate.
Biochem. Biophys. Res. Commun.
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The tellurium oxyanion tellurite is toxic to most organisms because of its ability to generate oxidative stress. However, the detailed mechanism(s) how this toxicant interferes with cellular processes have yet to be fully understood. As part of our effort to decipher the molecular interactions of tellurite with living systems, we have evaluated the global metabolism of ?-ketoglutarate a known antioxidant in Escherichia coli. Tellurite-exposed cells displayed reduced activity of the KG dehydrogenase complex (KGDHc), resulting in increased intracellular KG content. This complexs reduced activity seems to be due to decreased transcription in the stressed cells of sucA, a gene that encodes the E1 component of KGDHc. Furthermore, it was demonstrated that the increase in total reactive oxygen species and superoxide observed upon tellurite exposure was more evident in wild type cells than in E. coli with impaired KGDHc activity. These results indicate that KG may be playing a pivotal role in combating tellurite-mediated oxidative damage.
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Enhancing the antibiotic antibacterial effect by sub lethal tellurite concentrations: tellurite and cefotaxime act synergistically in Escherichia coli.
PLoS ONE
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The emergence of antibiotic-resistant pathogenic bacteria during the last decades has become a public health concern worldwide. Aiming to explore new alternatives to treat antibiotic-resistant bacteria and given that the tellurium oxyanion tellurite is highly toxic for most microorganisms, we evaluated the ability of sub lethal tellurite concentrations to strengthen the effect of several antibiotics. Tellurite, at nM or µM concentrations, increased importantly the toxicity of defined antibacterials. This was observed with both gram negative and gram positive bacteria, irrespective of the antibiotic or tellurite tolerance of the particular microorganism. The tellurite-mediated antibiotic-potentiating effect occurs in laboratory and clinical, uropathogenic Escherichia coli, especially with antibiotics disturbing the cell wall (ampicillin, cefotaxime) or protein synthesis (tetracycline, chloramphenicol, gentamicin). In particular, the effect of tellurite on the activity of the clinically-relevant, third-generation cephalosporin (cefotaxime), was evaluated. Cell viability assays showed that tellurite and cefotaxime act synergistically against E. coli. In conclusion, using tellurite like an adjuvant could be of great help to cope with several multi-resistant pathogens.
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Reduction of the monounsaturated fatty acid content of Escherichia coli results in increased resistance to oxidative damage.
Microbiology (Reading, Engl.)
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Reactive oxygen species (ROSs) affect several macromolecules and cellular components in eukaryotic and prokaryotic cells. In this work, the effect of various ROS-generating compounds on the Escherichia coli membrane was studied. Membrane fatty acid profiles, oxidative damage levels and bacterial resistance to these toxicants were determined. Studies included wild-type cells as well as a strain exhibiting a modified monounsaturated fatty acid (MUFA) profile (accomplished by overexpressing the ?-hydroxyacyl acyl carrier protein dehydratase-encoding gene, fabA). Levels of membrane MUFAs and oxidative damage markers decreased slightly upon toxicant exposure with a concomitant increase in cell resistance to these ROS-generating compounds. A direct relationship between MUFAs and lipid peroxidation was observed. The lower the MUFA the lower the peroxide levels, suggesting that MUFAs are targets for membrane lipid oxidation.
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Biomimetic, mild chemical synthesis of CdTe-GSH quantum dots with improved biocompatibility.
PLoS ONE
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Multiple applications of nanotechnology, especially those involving highly fluorescent nanoparticles (NPs) or quantum dots (QDs) have stimulated the research to develop simple, rapid and environmentally friendly protocols for synthesizing NPs exhibiting novel properties and increased biocompatibility. In this study, a simple protocol for the chemical synthesis of glutathione (GSH)-capped CdTe QDs (CdTe-GSH) resembling conditions found in biological systems is described. Using only CdCl(2), K(2)TeO(3) and GSH, highly fluorescent QDs were obtained under pH, temperature, buffer and oxygen conditions that allow microorganisms growth. These CdTe-GSH NPs displayed similar size, chemical composition, absorbance and fluorescence spectra and quantum yields as QDs synthesized using more complicated and expensive methods.CdTe QDs were not freely incorporated into eukaryotic cells thus favoring their biocompatibility and potential applications in biomedicine. In addition, NPs entry was facilitated by lipofectamine, resulting in intracellular fluorescence and a slight increase in cell death by necrosis. Toxicity of the as prepared CdTe QDs was lower than that observed with QDs produced by other chemical methods, probably as consequence of decreased levels of Cd(+2) and higher amounts of GSH. We present here the simplest, fast and economical method for CdTe QDs synthesis described to date. Also, this biomimetic protocol favors NPs biocompatibility and helps to establish the basis for the development of new, "greener" methods to synthesize cadmium-containing QDs.
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Effect of tellurite-mediated oxidative stress on the Escherichia coli glycolytic pathway.
Biometals
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To unveil the metabolic impact of tellurite in the bacterial cell, the effect of this toxicant on the expression and activity of key enzymes of the Escherichia coli glycolytic pathway was analyzed. E. coli exposure to tellurite results in: (i) increased glucose consumption, which was paralleled by an increased expression of the glucose transporter-encoding gene ptsG, (ii) augmented phosphoglucoisomerase activity and pgi transcription, (iii) decreased activity of the enzymatic regulators phosphofructokinase and pyruvate kinase. In spite of these observations, increased intracellular pyruvate, phosphoenol pyruvate and phosphorylated sugars was observed. E. coli lacking key glycolytic enzymes was considerably more sensitive to tellurite than the parental, isogenic, wild type strain. Taken together, these results suggest that increasing the availability of key metabolites (pyruvate, phosphoenol pyruvate, NADPH), required to respond to tellurite mediated-stress, E. coli shifts the carbon flux towards the pentose phosphate pathway thus facilitating the functioning of the Entner-Doudoroff pathway and/or the glycolytic productive phase.
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