Biomineralization of lithium nanoparticles by Li-resistant Pseudomonas rodhesiae isolated from the Atacama salt flat.

BACKGROUND: The Atacama salt flat is located in northern Chile, at 2300 m above sea level, and has a high concentration of lithium, being one of the main extraction sites in the world. The effect of lithium on microorganism communities inhabiting environments with high concentrations of this metal has been scarcely studied. A few works have studied the microorganisms present in lithium-rich salt flats (Uyuni and Hombre Muerto in Bolivia and Argentina, respectively). Nanocrystals formation through biological mineralization has been described as an alternative for microorganisms living in metal-rich environments to cope with metal ions. However, bacterial lithium biomineralization of lithium nanostructures has not been published to date. In the present work, we studied lithium-rich soils of the Atacama salt flat and reported for the first time the biological synthesis of Li nanoparticles. RESULTS:  Bacterial communities were evaluated and a high abundance of Cellulomonas, Arcticibacter, Mucilaginibacter, and Pseudomonas were determined. Three lithium resistant strains corresponding to Pseudomonas rodhesiae, Planomicrobium koreense, and Pseudomonas sp. were isolated (MIC > 700 mM). High levels of S2- were detected in the headspace of P. rodhesiae and Pseudomonas sp. cultures exposed to cysteine. Accordingly, biomineralization of lithium sulfide-containing nanomaterials was determined in P. rodhesiae exposed to lithium salts and cysteine. Transmission electron microscopy (TEM) analysis of ultrathin sections of P. rodhesiae cells biomineralizing lithium revealed the presence of nanometric materials. Lithium sulfide-containing nanomaterials were purified, and their size and shape determined by dynamic light scattering and TEM. Spherical nanoparticles with an average size < 40 nm and a hydrodynamic size ~ 44.62 nm were determined. CONCLUSIONS: We characterized the bacterial communities inhabiting Li-rich extreme environments and reported for the first time the biomineralization of Li-containing nanomaterials by Li-resistant bacteria. The biosynthesis method described in this report could be used to recover lithium from waste batteries and thus provide a solution to the accumulation of batteries.

Biomineralization of lithium nanoparticles by Li-resistant Pseudomonas rodhesiae isolated from the Atacama salt flat

BACKGROUND: The Atacama salt flat is located in northern Chile, at 2300 m above sea level, and has a high concentration of lithium, being one of the main extraction sites in the world. The effect of lithium on microorganism communities inhabiting environments with high concentrations of this metal has been scarcely studied. A few works have studied the microorganisms present in lithium-rich salt flats (Uyuni and Hombre Muerto in Bolivia and Argentina, respectively). Nanocrystals formation through biological mineralization has been described as an alternative for microorganisms living in metal-rich environments to cope with metal ions. However, bacterial lithium biomineralization of lithium nanostructures has not been published to date. In the present work, we studied lithium-rich soils of the Atacama salt flat and reported for the first time the biological synthesis of Li nanoparticles. RESULTS: Bacterial communities were evaluated and a high abundance of Cellulomonas, Arcticibacter, Mucilaginibacter, and Pseudomonas were determined. Three lithium resistant strains corresponding to Pseudomonas rodhesiae, Planomicrobium koreense, and Pseudomonas sp. were isolated (MIC > 700 mM). High levels of S2− were detected in the headspace of P. rodhesiae and Pseudomonas sp. cultures exposed to cysteine. Accordingly, biomineralization of lithium sulfide-containing nanomaterials was determined in P. rodhesiae exposed to lithium salts and cysteine. Transmission electron microscopy (TEM) analysis of ultrathin sections of P. rodhesiae cells biomineralizing lithium revealed the presence of nanometric materials. Lithium sulfide-containing nanomaterials were purified, and their size and shape determined by dynamic light scattering and TEM. Spherical nanoparticles with an average size < 40 nm and a hydro-dynamic size ~ 44.62 nm were determined. CONCLUSIONS: We characterized the bacterial communities inhabiting Li-rich extreme environments and reported for the first time the biomineralization of Li-containing nanomaterials by Li-resistant bacteria. The biosynthesis method described in this report could be used to recover lithium from waste batteries and thus provide a solution to the accumulation of batteries.

Production of cadmium sulfide quantum dots by the lithobiontic Antarctic strain Pedobacter sp. UYP1 and their application as photosensitizer in solar cells.

BACKGROUND: Microbes are present in almost every environment on Earth, even in those with extreme environmental conditions such as Antarctica, where rocks may represent the main refuge for life. Lithobiontic communities are composed of microorganisms capable of colonizing rocks and, as it is a not so well studied bacterial community, they may represent a very interesting source of diversity and functional traits with potential for biotechnological applications. In this work we analyzed the ability of Antarctic lithobiontic bacterium to synthesize cadmium sulfide quantum dots (CdS QDs) and their potential application in solar cells. RESULTS: A basaltic andesite rock sample was collected from Fildes Peninsula, King George Island, Antarctica, and processed in order to isolate lithobiontic bacterial strains. Out of the 11 selected isolates, strain UYP1, identified as Pedobacter, was chosen for further characterization and analysis due to its high cadmium tolerance. A protocol for the biosynthesis of CdS QDs was developed and optimized for this strain. After 20 and 80 min of synthesis, yellow-green and orange-red fluorescent emissions were observed under UV light, respectively. QDs were characterized through spectroscopic techniques, dynamic light scattering analysis, high-resolution transmission electron microscopy and energy dispersive x-ray spectroscopy. Nanostructures of 3.07 nm, composed of 51.1% cadmium and 48.9% sulfide were obtained and further used as photosensitizer material in solar cells. These solar cells were able to conduct electrons and displayed an open circuit voltage of 162 mV, a short circuit current density of 0.0110 mA cm-2, and had an efficiency of conversion up to 0.0016%, which is comparable with data previously reported for solar cells sensitized with biologically produced quantum dots. CONCLUSIONS: We report a cheap, rapid and eco-friendly protocol for the production of CdS QDs by an Antarctic lithobiontic bacterium, Pedobacter, a genus that was not previously reported as a quantum dot producer. The application of the biosynthesized QDs as sensitizer material in solar cells was validated.

Bacterial phototoxicity of biomimetic CdTe-GSH quantum dots.

AIM: Fluorescent semiconductor nanoparticles or quantum dots (QDs) have excellent properties as photosensitizers in photodynamic therapy. This is mainly a consequence of their nanometric size and the generation of light-activated redox species. In previous works, we have reported the low-cost biomimetic synthesis of glutathione (GSH) capped QDs (CdTe-GSH QDs) with high biocompatibility. However, no studies have been performed to determine their phototoxic effect. The aim of this work was to characterize the light-induced toxicity of green (QDs500 ) and red (QDs600 ) QDs in Escherichia coli, and to study the molecular mechanism involved. METHODS AND RESULTS: Photodegradation and reduction power of biomimetic QDs was determined to analyse their potential for radical generation. Escherichia coli cells were exposed to photoactivated QDs and viability was evaluated at different times. High toxicity was determined in E. coli cells exposed to photoactivated QDs, particularly QDs500 . The molecular mechanism involved in QDs phototoxicity was studied by determining Cd2+ -release and intracellular reactive oxygen species (ROS). Cells exposed to photoactivated QDs500 presented high levels of ROS. Cells exposed to photoactivated QDs500 presented high levels of ROS. Finally, to understand this phenomenon and the importance of oxidative and cadmium-stress in QDs-mediated phototoxicity, experiments were performed in E. coli mutants in ROS and Cd2+ response genes. As expected, E. coli mutants in ROS response genes were more sensitive than the wt strain to photoactivated QDs, with a higher effect in green-QDs500 . No increase in phototoxicity was observed in cadmium-related mutants. CONCLUSION: Obtained results indicate that light exposure increases the toxicity of biomimetic QDs on E. coli cells. The mechanism of bacterial phototoxicity of biomimetic CdTe-GSH QDs is mostly associated with ROS generation. SIGNIFICANCE AND IMPACT OF THE STUDY: The results presented establish biomimetic CdTe-GSH QDs as a promising cost-effective alternative against microbial infections, particularly QDs500 .

Synthesis of salt-stable fluorescent nanoparticles (quantum dots) by polyextremophile halophilic bacteria.

Here we report the biological synthesis of CdS fluorescent nanoparticles (Quantum Dots, QDs) by polyextremophile halophilic bacteria isolated from Atacama Salt Flat (Chile), Uyuni Salt Flat (Bolivia) and the Dead Sea (Israel). In particular, a Halobacillus sp. DS2, a strain presenting high resistance to NaCl (3-22%), acidic pH (1-4) and cadmium (CdCl2 MIC: 1,375 mM) was used for QDs biosynthesis studies. Halobacillus sp. synthesize CdS QDs in presence of high NaCl concentrations in a process related with their capacity to generate S2- in these conditions. Biosynthesized QDs were purified, characterized and their stability at different NaCl concentrations determined. Hexagonal nanoparticles with highly defined structures (hexagonal phase), monodisperse size distribution (2-5 nm) and composed by CdS, NaCl and cysteine were determined by TEM, EDX, HRXPS and FTIR. In addition, QDs biosynthesized by Halobacillus sp. DS2 displayed increased tolerance to NaCl when compared to QDs produced chemically or biosynthesized by non-halophilic bacteria. This is the first report of biological synthesis of salt-stable QDs and confirms the potential of using extremophile microorganisms to produce novel nanoparticles. Obtained results constitute a new alternative to improve QDs properties, and as consequence, to increase their industrial and biomedical applications.

A comparative analysis of tellurite detoxification by members of the genus Shewanella.

The increasing industrial utilization of tellurium has resulted in an important environmental pollution with the soluble, extremely toxic oxyanion tellurite. In this context, the use of microorganisms for detoxifying tellurite or tellurium biorecovery has gained great interest. The ability of different Shewanella strains to reduce tellurite to elemental tellurium was assessed; the results showed that the reduction process is dependent on electron transport and the ∆pH gradient. While S. baltica OS155 showed the highest tellurite resistance, S. putrefaciens was the most efficient in reducing tellurite. Moreover, pH-dependent tellurite transformation was associated with tellurium precipitation as tellurium dioxide. In summary, this work highlights the high tellurite reduction/detoxification ability exhibited by a number of Shewanella species, which could represent the starting point to develop friendly methods for the recovery of elemental tellurium (or tellurium dioxide).

"Use of acidophilic bacteria of the genus Acidithiobacillus to biosynthesize CdS fluorescent nanoparticles (quantum dots) with high tolerance to acidic pH".

The use of bacterial cells to produce fluorescent semiconductor nanoparticles (quantum dots, QDs) represents a green alternative with promising economic potential. In the present work, we report for the first time the biosynthesis of CdS QDs by acidophilic bacteria of the Acidithiobacillus genus. CdS QDs were obtained by exposing A. ferrooxidans, A. thiooxidans and A. caldus cells to sublethal Cd2+ concentrations in the presence of cysteine and glutathione. The fluorescence of cadmium-exposed cells moves from green to red with incubation time, a characteristic property of QDs associated with nanocrystals growth. Biosynthesized nanoparticles (NPs) display an absorption peak at 360nm and a broad emission spectra between 450 and 650nm when excited at 370nm, both characteristic of CdS QDs. Average sizes of 6 and 10nm were determined for green and red NPs, respectively. The importance of cysteine and glutathione on QDs biosynthesis in Acidithiobacillus was related with the generation of H2S. Interestingly, QDs produced by acidophilic bacteria display high tolerance to acidic pH. Absorbance and fluorescence properties of QDs was not affected at pH 2.0, a condition that totally inhibits the fluorescence of QDs produced chemically or biosynthesized by mesophilic bacteria (stable until pH 4.5-5.0). Results presented here constitute the first report of the generation of QDs with improved properties by using extremophile microorganisms.

Pigments from UV-resistant Antarctic bacteria as photosensitizers in Dye Sensitized Solar Cells.

Here we report the use of pigments produced by UV-resistant Antarctic bacteria as photosensitizers in Dye Sensitized Solar Cells (DSSCs). Pigments were obtained from red and yellow colored psychrotolerant bacteria isolated from soils of King George Island, Antarctica. Based on metabolic characteristics and 16s DNA sequence, pigmented bacteria were identified as Hymenobacter sp. (red) and Chryseobacterium sp. (yellow). Pigments produced by these microorganisms were extracted and classified as carotenoids based on their spectroscopic and structural characteristics, determined by UV-Vis spectrophotometry and infrared spectroscopy (FTIR), respectively. With the purpose of develop green solar cells based on bacterial pigments, the photostability and capacity of these molecules as light harvesters in DSSCs were determined. Absorbance decay assays determined that bacterial carotenoids present high photostability. In addition, solar cells based on these photosensitizers exhibit an open circuit voltage (VOC) of 435.0 [mV] and a short circuit current density (ISC) of 0.2 [mA·cm(-2)] for the red pigment, and a VOC of 548.8 [mV] and a ISC of 0.13 [mA·cm(-2)] for the yellow pigment. This work constitutes the first approximation of the use of pigments produced by non-photosynthetic bacteria as photosensitizers in DSSCs. Determined photochemical characteristics of bacterial pigments, summed to their easy obtention and low costs, validates its application as photosensitizers in next-generation biological solar cells.

Biological synthesis of fluorescent nanoparticles by cadmium and tellurite resistant Antarctic bacteria: exploring novel natural nanofactories.

BACKGROUND: Fluorescent nanoparticles or quantum dots (QDs) have been intensely studied for basic and applied research due to their unique size-dependent properties. There is an increasing interest in developing ecofriendly methods to synthesize these nanoparticles since they improve biocompatibility and avoid the generation of toxic byproducts. The use of biological systems, particularly prokaryotes, has emerged as a promising alternative. Recent studies indicate that QDs biosynthesis is related to factors such as cellular redox status and antioxidant defenses. Based on this, the mixture of extreme conditions of Antarctica would allow the development of natural QDs producing bacteria. RESULTS: In this study we isolated and characterized cadmium and tellurite resistant Antarctic bacteria capable of synthesizing CdS and CdTe QDs when exposed to these oxidizing heavy metals. A time dependent change in fluorescence emission color, moving from green to red, was determined on bacterial cells exposed to metals. Biosynthesis was observed in cells grown at different temperatures and high metal concentrations. Electron microscopy analysis of treated cells revealed nanometric electron-dense elements and structures resembling membrane vesicles mostly associated to periplasmic space. Purified biosynthesized QDs displayed broad absorption and emission spectra characteristic of biogenic Cd nanoparticles. CONCLUSIONS: Our work presents a novel and simple biological approach to produce QDs at room temperature by using heavy metal resistant Antarctic bacteria, highlighting the unique properties of these microorganisms as potent natural producers of nano-scale materials and promising candidates for bioremediation purposes.

The Porphyromonas gingivalis O antigen is required for inhibition of apoptosis in gingival epithelial cells following bacterial infection.

BACKGROUND AND OBJECTIVE: Porphyromonas gingivalis infection induces apoptosis inhibition in gingival epithelial cells; however, it is not fully understood which bacterial effectors are involved in this process. The aim of this study is to evaluate whether the P. gingivalis lipopolysaccharide (LPS), specifically the O-antigen region, affects adherence, invasion, viability and apoptosis of gingival epithelial cells. MATERIAL AND METHODS: Gingival epithelial cells (OKF6/TERT2 line) were infected by different freshly prepared P. gingivalis clinical isolates, obtained from subjects with chronic periodontitis (CP3 and CP4) and healthy individuals (H1 and H3). Periodontitis and healthy isolates show differences in O-antigen production, as healthy isolates lack the O-antigen region. In addition, cells were infected by a site-specific mutant lacking the O-antigen portion. After 24 h postinfection, cell proliferation, viability and apoptosis were evaluated by Trypan blue, MTS and annexin V assays, respectively. Bacterial invasion, adhesion and proliferation were measured by gentamicin/metronidazole protection assays. Finally, toll-like receptor (TLR)2 and TLR4 mRNA expression was evaluated by quantitative reverse transcription-polymerase chain reaction. Statistical analysis was performed using ANOVA, Tukey's or Dunnett's tests (p < 0.05). RESULTS: At 24 h postinfection, strains lacking the O-antigen region (healthy isolates and O-antigen ligase-deficient strain) were unable to increase proliferation and viability, or decrease apoptosis as compared with strains producing intact LPS (periodontitis isolates and reference strain). However, the presence of the O-antigen neither contributed to changes in the ability of the bacteria to adhere to or invade cells, nor to intracellular survival. The presence of O-antigen also increased the expression of TLR4 (nearly sixfold), which correlated with inhibition of apoptosis. CONCLUSION: The O-antigen region of P. gingivalis LPS is required to increase gingival epithelial cell viability upon infection by bacteria and this increase is attributable to a reduction in apoptosis. Moreover, although bacterial internalization is required, the effects observed are not due to alterations in P. gingivalis adherence, invasion or intracellular survival. Interestingly, inhibition of apoptosis correlates with increased TLR4 expression, suggesting a role for TLR4 in this process.

Low-temperature biosynthesis of fluorescent semiconductor nanoparticles (CdS) by oxidative stress resistant Antarctic bacteria.

Bacterial biosynthesis of nanoparticles represents a green alternative for the production of nanostructures with novel properties. Recently, the importance of antioxidant molecules on the biosynthesis of semiconductor fluorescent nanoparticles (quantum dots, QDs) by mesophilic bacteria was reported. The objective of this work was the isolation of psychrotolerant, oxidative stress-resistant bacteria from Antarctica to determine their ability for biosynthesizing CdS QDs at low temperatures. QDs biosynthesis at 15 °C was evaluated by determining their spectroscopic properties after exposing oxidative-stress resistant isolates identified as Pseudomonas spp. to Cd(2+) salts. To characterize the QDs biosynthetic process, the effect of metal exposure on bacterial fluorescence was determined at different times. Time-dependent changes in fluorescence color (green to red), characteristic of QDs, were observed. Electron microscopy analysis of fluorescent cells revealed that biosynthesized nanometric structures localize at the cell periphery. QDs were purified from the bacterial isolates and their fluorescence properties were characterized. Emission spectra displayed classical CdS peaks when excited with UV light. Thiol content, peroxidase activity, lipopolysaccharide synthesis, metabolic profiles and sulfide generation were determined in QDs-producing isolates. No relationship between QDs production and cellular thiol content or peroxidase activity was found. However, sulfide production enhanced CdS QDs biosynthesis. In this work, the use of Antarctic psychrotolerant Pseudomonas spp. for QDs biosynthesis at low temperature is reported for the first time.

Quantum dot-based assay for Cu(2+) quantification in bacterial cell culture.

A simple and sensitive method for quantification of nanomolar copper with a detection limit of 1.2×10(-10)M and a linear range from 10(-9) to 10(-8)M is reported. For the most useful analytical concentration of quantum dots, 1160µg/ml, a 1/Ksv value of 11µM Cu(2+) was determined. The method is based on the interaction of Cu(2+) with glutathione-capped CdTe quantum dots (CdTe-GSH QDs) synthesized by a simple and economic biomimetic method. Green CdTe-GSH QDs displayed the best performance in copper quantification when QDs of different sizes/colors were tested. Cu(2+) quantification is highly selective given that no significant interference of QDs with 19 ions was observed. No significant effects on Cu(2+) quantification were determined when different reaction matrices such as distilled water, tap water, and different bacterial growth media were tested. The method was used to determine copper uptake kinetics on Escherichia coli cultures. QD-based quantification of copper on bacterial supernatants was compared with atomic absorption spectroscopy as a means of confirming the accuracy of the reported method. The mechanism of Cu(2+)-mediated QD fluorescence quenching was associated with nanoparticle decomposition.