Phosphate-Arsenic Interactions in Halophilic Microorganisms of the Microbial Mat from Laguna Tebenquiche: from the Microenvironment to the Genomes.

Arsenic (As) is a metalloid present in the earth's crust and widely distributed in the environment. Due to its high concentrations in the Andean valleys and its chemical similarity with phosphorus (P), its biological role in Andean Microbial Ecosystems (AMEs) has begun to be studied. The AMEs are home to extremophilic microbial communities that form microbial mats, evaporites, and microbialites inhabiting Andean lakes, puquios, or salt flats. In this work, we characterize the biological role of As and the effect of phosphate in AMEs from the Laguna Tebenquiche (Atacama Desert, Chile). Using micro X-ray fluorescence, the distribution of As in microbial mat samples was mapped. Taxonomic and inferred functional profiles were obtained from enriched cultures of microbial mats incubated under As stress and different phosphate conditions. Additionally, representative microorganisms highly resistant to As and able to grow under low phosphate concentration were isolated and studied physiologically. Finally, the genomes of the isolated Salicola sp. and Halorubrum sp. were sequenced to analyze genes related to both phosphate metabolism and As resistance. The results revealed As as a key component of the microbial mat ecosystem: (i) As was distributed across all sections of the microbial mat and represented a significant weight percentage of the mat (0.17 %) in comparison with P (0.40%); (ii) Low phosphate concentration drastically changed the microbial community in microbial mat samples incubated under high salinity and high As concentrations; (iii) Archaea and Bacteria isolated from the microbial mat were highly resistant to arsenate (up to 500 mM), even under low phosphate concentration; (iv) The genomes of the two isolates were predicted to contain key genes in As metabolism (aioAB and arsC/acr3) and the genes predicted to encode the phosphate-specific transport operon (pstSCAB-phoU) are next to the arsC gene, suggesting a functional relationship between these two elements.

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.