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.

Expression of the ubiE gene of Geobacillus stearothermophilus V in Escherichia coli K-12 mediates the evolution of selenium compounds into the headspace of selenite- and selenate-amended cultures.

The ubiE gene of Geobacillus stearothermophilus V, with its own promoter, was cloned and introduced into Escherichia coli. The cloned gene complemented the ubiE gene deficiency of E. coli AN70. In addition, the expression of this gene in E. coli JM109 resulted in the evolution of volatile selenium compounds when these cells were grown in selenite- or selenate-amended media. These compounds were dimethyl selenide and dimethyl diselenide.

Fate of selenate and selenite metabolized by Rhodobacter sphaeroides.

Cultures of a purple nonsulfur bacterium, Rhodobacter sphaeroides, amended with approximately 1 or approximately 100 ppm selenate or selenite, were grown phototrophically to stationary phase. Analyses of culture headspace, separated cells, and filtered culture supernatant were carried out using gas chromatography, X-ray absorption spectroscopy, and inductively coupled plasma spectroscopy-mass spectrometry, respectively. While selenium-amended cultures showed much higher amounts of SeO(3)(2-) bioconversion than did analogous selenate experiments (94% uptake for SeO(3)(2-) as compared to 9.6% for SeO(4)(2-)-amended cultures from 100-ppm solutions), the chemical forms of selenium in the microbial cells were not very different except at exposure to high concentrations of selenite. Volatilization accounted for only a very small portion of the accumulated selenium; most was present in organic forms and the red elemental form.

X-ray absorption spectroscopy of selenium-containing amino acids.

The selenium K-edge X-ray absorption spectra of selenomethionine, selenocysteine, selenocystine, and sulfo-selenocystine in solution are compared with the corresponding sulfur K-edge spectra of the sulfur analogues of these compounds. The selenium and sulfur spectra follow similar trends, although the latter are significantly sharper owing to the longer core hole lifetime at the lower energies where sulfur absorbs. The spectra of the selenium compounds are sufficiently distinct that it is reasonable to expect that curve fitting will allow the speciation of the forms of selenium in complex biological samples.