Cytological aspects of resistance to potassium tellurite conferred on Pseudomonas cells by plasmids

The ultrastructure of strains Pseudomonas putida BS228 and Pseudomonas aeruginosa ML4262 harboring plasmids pBS10, pBS31, and pBS221, which determine resistance to potassium tellurite, was studied. Bacteria were grown in media containing increasing concentrations of potassium tellurite. Crystalline structures containing tellurium appeared in their periplasmic space. The dynamics of crystal growth was studied. Crystals were released into the medium by pinching off of the outer membrane vesicles containing growing crystals. A possible mechanism of this process was described; cytobiochemical peculiarities were discussed.     

Cysteine metabolism-related genes and bacterial resistance to potassium tellurite

Tellurite exerts a deleterious effect on a number of small molecules containing sulfur moieties that have a recognized role in cellular oxidative stress. Because cysteine is involved in the biosynthesis of glutathione and other sulfur-containing compounds, we investigated the expression of Geobacillus stearothermophilus V cysteine-related genes cobA, cysK, and iscS and Escherichia coli cysteine regulon genes under conditions that included the addition of K₂TeO₃ to the culture medium. Results showed that cell tolerance to tellurite correlates with the expression level of the cysteine metabolic genes and that these genes are up-regulated when tellurite is present in the growth medium.     

Selective erythrocyte potassium efflux following pulse treatment with tellurite.

Human erythrocytes exposed to 0.1 mM tellurite (K2TeO3) in an isotonic buffered choline chloride medium for 15 min at 37 degrees C, washed, and incubated further in the absence of the chemical in the buffer, exhibited selective leakiness for potassium within minutes. The potassium efflux curve was sigmoidal, with an initially slow leakage followed by a sharp rise (first-order kinetics) and a plateau by 60 min. After 15 min, 30-50% of the total potassium concentration had leaked from the cells, although less than 1% lysis had occurred. The control cells incubated in buffer with no K2TeO3 exhibited no potassium leakage. The mean volume of the K2TeO3-treated erythrocytes increased and their median density decreased, indicating changes in the colloid osmotic state and physical characteristics of the cells. However, cells pretreated with K2TeO3 exhibited no significant change in glutathione (GSH) concentration and no membrane lipid peroxidation, unlike cells pretreated with t-butylhydroperoxide (Deuticke et al., Biochim. Bio phys. Acta, 899, 125-128, 1987). The enhanced potassium permeability of the K2TeO3-treated erythrocytes preceded the increase in cell volume, intracellular hydration, and a decrease in median density. We suggest that perturbation of the lipid-protein interaction in the membrane by the oxidant alters the potassium permeability and results in the selective leakage with eventual hemolysis.     

High level, intrinsic resistance of Natronococcus occultus to potassium tellurite

Natronococcus occultus, a haloalkaliphilic archaeon, was examined for its resistance to potassium tellurite. Cells grown in the presence of 1 mM potassium tellurite reduced it to metallic tellurium resulting in the deposition of intracellular crystals in the cytoplasm. The minimal inhibitory concentration for potassium tellurite was 10 mM. N. occultus had an inducible tellurite reductase activity. Cell-free extracts catalyzed the enzymatic reduction of potassium tellurite in a reaction which was dependent on NADH oxidation and a reduced environment.     

Reduction of potassium tellurite to elemental tellurium and its effect on the plasma membrane redox components of the facultative phototroph Rhodobacter capsulatus

Summary.  Anaerobically light-grown cells of Rhodobacter capsulatus B100 are highly resistant to the toxic oxyanion tellurite (TeO₃ ²⁻; minimal inhibitory concentration, 250 μg/ml). This study examines, for the first time, some structural and biochemical features of cells and plasma membrane fragments of this facultative phototroph grown in the presence of 50μg of K₂TeO₃ per ml. Through the use of transmission microscopy and X-ray microanalysis we show that several “needlelike” shaped granules of elemental tellurium are accumulated into the cytosol near the intracytoplasmic membrane system. Flash-spectroscopy, oxygen consumption measurements, and difference spectra analysis indicated that membrane vesicles (chromatophores) isolated from tellurite-grown cells are able to catalyze both photosynthetic and respiratory electron transport activities, although they are characterized by a low c-type cytochrome content (mostly soluble cytochrome c ₂). This feature is paralleled by a low cytochrome c oxidase activity and with an NADH-dependent respiration which is catalyzed by a pathway leading to a quinol oxidase (Qox) inhibited by high (millimolar) concentrations of cyanide (CN⁻). Conversely, membranes from R. capsulatus B100 cells grown in the absence of tellurite are characterized by a branched respiratory chain in which the cytochrome c oxidase pathway (blocked by CN⁻ in the micromolar range) accounts for 35–40% of the total NADH-dependent oxygen consumption, while the remaining activity is catalyzed by the quinol oxidase pathway. These data have been interpreted to show that tellurite resistance of R. capsulatus B100 is characterized by the presence of a modified plasma-membrane-associated electron transport system. Received May 2, 2002; accepted July 26, 2002; published online May 21, 2003 RID="*" ID="*" Correspondence and reprints: Department of Biology, University of Bologna, Via Irnerio 42, 40126 Bologna, Italy.     

The toxicity of potassium tellurite to Staphylococcus aureus in rabbit plasma fibrinogen agar

The potassium tellurite concentration, 0.01% w/v, in Baird-Parker agar has been recommended for plasma coagulase media such as pig or rabbit fibrinogen agar. Comparative tests have shown that with some strains of Staphylococcus aureus this level of potassium tellurite is too inhibitory and it should be reduced four-fold to 0.0025% w/v to maximize the isolation rate. It is postulated that egg yolk in Baird-Parker agar has a protective effect on staphylococci against the inhibitory action of tellurite.     

The toxicity of potassium tellurite to Staphylococcus aureus in rabbit plasma fibrinogen agar

The potassium tellurite concentration, 0.01% w/v, in Baird-Parker agar has been recommended for plasma coagulase media such as pig or rabbit fibrinogen agar. Comparative tests have shown that with some strains of Staphylococcus aureus this level of potassium tellurite is too inhibitory and it should be reduced four-fold to 0.0025% w/v to maximize the isolation rate. It is postulated that egg yolk in Baird-Parker agar has a protective effect on staphylococci against the inhibitory action of tellurite.     

Selenite and tellurite reduction by Shewanella oneidensis

Shewanella oneidensis MR-1 reduces selenite and tellurite preferentially under anaerobic conditions. The Se(0) and Te(0) deposits are located extracellularly and intracellularly, respectively. This difference in localization and the distinct effect of some inhibitors and electron acceptors on these reduction processes are taken as evidence of two independent pathways.     

Glutathione is a target in tellurite toxicity and is protected by tellurite resistance determinants in Escherichia coli

Tellurite (TeO3(2-)) is highly toxic to most microorganisms. The mechanisms of toxicity or resistance are poorly understood. It has been shown that tellurite rapidly depletes the reduced thiol content within wild-type Escherichia coli. We have shown that the presence of plasmid-borne tellurite-resistance determinants protects against general thiol oxidation by tellurite. In the present study we observe that the tellurite-dependent depletion of cellular thiols in mutants of the glutathione and thioredoxin thiol:redox system was less than in wild-type cells. To identify the type of low-molecular-weight thiol compounds affected by tellurite exposure, the thiol-containing molecules were analyzed by reverse phase HPLC as their monobromobimane derivatives. Results indicated that reduced glutathione is a major initial target of tellurite reactivity within the cell. Other thiol species are also targeted by tellurite, including reduced coenzyme A. The presence of the tellurite resistance determinants kilA and ter protect against the loss of reduced glutathione by as much as 60% over a 2 h exposure. This protection of glutathione oxidation is likely key to the resistance mechanism of these determinants. Additionally, the thiol oxidation response curves were compared between selenite and tellurite. The loss of thiol compounds within the cell recovered from selenite but not to tellurite.     

Quinone-mediated reduction of selenite and tellurite by Escherichia coli

The reduction of selenite (Se(IV)) and tellurite (Te(IV)) by Escherichia coli was significantly enhanced by various quinone redox mediators (lawsone, menadione, anthraquinone-2-sulfonate, and anthraquinone-2,6-disulfonate). In the presence of 0.2mM lawsone, over 99.1% Se(IV) and around 96.4% Te(IV) were reduced in 8 h, at average reduction rates of 9.1 and 7.6 mM g cell(-1) h(-1), respectively. Better mediated reduction of Se(IV) and Te(IV) were observed when lawsone concentration increased from 0.1 to 0.4 mM and cell concentration increased from 0.1 to 0.6 g l(-1), respectively. Transmission electron microscopy analysis revealed the formation of both intracellular and extracellular Se(0) nanospheres or Te(0) nanorods, and the presence of lawsone increased the formation and accumulation of extracellular precipitates. The efficient mediated microbial reduction of Se(IV)/Te(IV) may be exploited for pollution removal and biological nanomaterials production.