Accumulation Of Arsenate Research Articles

Accumulation and mobilization of arsenate by Fe(III) polyions trapped in a Ca-polygalacturonate network

The role of Fe(III) stored at the soil-root interface in the accumulation of arsenate and the influence of citric acid on the As(V) mobility were investigated by using Ca-polygalacturonate networks (PGA). The results indicate that in the 2.5-6.2 pH range Fe(III) interacts with As(V) leading to the sorption of As(V) on Fe(III) precipitates or Fe-As coprecipitates. The FT-IR analysis of these precipitates evidenced that the interaction produces Fe(III)-As(V) inner-sphere complexes with either monodentate or bidentate binuclear attachment of As(V) depending on pH. In the 3.0-6.0 pH range, As(V) diffuses freely through the polysaccharidic matrix that was found to exert a negligible reducing action towards As(V). At pH 6.0 citric acid is able to mobilize arsenate from the As-Fe-PGA network through the complexation of the Fe(III) polyions that leads to the release of As(V).

Enzyme phylogenies as markers for the oxidation state of the environment: The case of respiratory arsenate reductase and related enzymes

BackgroundPhylogenies of certain bioenergetic enzymes have proved to be useful tools for deducing evolutionary ancestry of bioenergetic pathways and their relationship to geochemical parameters of the environment. Our previous phylogenetic analysis of arsenite oxidase, the molybdopterin enzyme responsible for the biological oxidation of arsenite to arsenate, indicated its probable emergence prior to the Archaea/Bacteria split more than 3 billion years ago, in line with the geochemical fact that arsenite was present in biological habitats on the early Earth. Respiratory arsenate reductase (Arr), another molybdopterin enzyme involved in microbial arsenic metabolism, serves as terminal oxidase, and is thus situated at the opposite end of bioenergetic electron transfer chains as compared to arsenite oxidase. The evolutionary history of the Arr-enzyme has not been studied in detail so far.ResultsWe performed a genomic search of genes related to arrA coding for the molybdopterin subunit. The multiple alignment of the retrieved sequences served to reconstruct a neighbor-joining phylogeny of Arr and closely related enzymes. Our analysis confirmed the previously proposed proximity of Arr to the cluster of polysulfide/thiosulfate reductases but also unravels a hitherto unrecognized clade even more closely related to Arr. The obtained phylogeny strongly suggests that Arr originated after the Bacteria/Archaea divergence in the domain Bacteria, and was subsequently laterally distributed within this domain. It further more indicates that, as a result of accumulation of arsenate in the environment, an enzyme related to polysulfide reductase and not to arsenite oxidase has evolved into Arr.ConclusionThese findings are paleogeochemically rationalized by the fact that the accumulation of arsenate over arsenite required the increase in oxidation state of the environment brought about by oxygenic photosynthesis.

Arsenic uptake and transport of Pteris vittata L. as influenced by phosphate and inorganic arsenic species under sand culture

In order to understand the similarity or difference of inorganic As species uptake and transport related to phosphorus in Ashyperaccumulator, uptake and transport of arsenate (As(V)) and arsenite (As(III)) were studied using Pteris vittata L. under sand culture. Higher concentrations of phosphate were found to inhibit accumulation of arsenate and arsenite in the fronds of P. vittata. The reduction in As accumulation was greater in old fronds than in young fronds, and relatively weak in root and rhizome. Moderate increases, from 0.05 to 0.3 mmol/L, in phosphate reduced uptake of As(III) more than As(V), while the reverse was observed at high concentrations of phosphate (≥1.0 mmol/L). Phosphate apparently reduced As transport and the proportion of As accumulated in fronds of P. vittata when As was supplied as As(V). It may in part be due to competition between phosphorus and As(V) during transport. In contrast, phosphate had a much smaller effect on As transport when the As was supplied as As(III). Therefore, the results from present experiments indicates that a higher concentration of phosphate suppressed As accumulation and transport in P. vittata, especially in the fronds, when exposed to As(V); but the suppression of phosphate to As transport may be insignificant when P. vittata exposed to As(III) under sand culture conditions. The finding will help to understand the interaction of P and As during their uptake process in P. vittata.

Enhanced Accumulation of Arsenate in Carp in the Presence of Titanium Dioxide Nanoparticles

In this study adsorption of arsenic (As) onto TiO2 nanoparticles and the facilitated transport of As into carp (Cyprinus carpio) by TiO2 nanoparticles was examined. Adsorption kinetics and adsorption isotherm were conducted by adding As(V) to TiO2 suspensions. Facilitated transport of As by TiO2 nanoparticles was assessed by accumulation tests exposing carp to As(V) contaminated water in the presence of TiO2 nanoparticles. The results showed that TiO2 nanoparticles had a significant adsorption capacity for As(V). Equilibrium was established within 30 min and the isotherm data was described by Freundlich isotherm. The KF and 1/n were 20.71 mg/g and 0.58, respectively. When exposed to As(V)-contaminated water in the presence of TiO2 nanoparticles, carp accumulated considerably more As, and As concentration in carp increased by 132% after 25 days exposure. Considerable As and TiO2 accumulated in intestine, stomach and gills of the fish, and the lowest level of accumulation was found in muscle. Accumulation of As and TiO2 in stomach, intestine and gills are significant. Arsenic accumulation in these tissues was enhanced by the presence of TiO2 nanoparticles. TiO2 nanoparticles that have accumulated in intestine and gills may release adsorbed As and As bound on TiO2 nanoparticles which cannot be released maybe transported by TiO2 nanoparticles as they transferred in the body. In this work, an enhancement of 80% and 126% As concentration in liver and muscle after 20 days of exposure was found.

Uptake, accumulation and depuration of sodium perchlorate and sodium arsenate in zebrafish (Danio rerio)

In toxicokinetics studies, interactions between chemicals in mixtures has been largely neglected. This study examines a mixture of perchlorate and arsenate because (1) they have the potential to co-occur in contaminated aquatic habitats, and (2) a previous study by the authors found possible toxicological interactive effects. In the present study, zebrafish (Danio rerio) were exposed to two concentrations of sodium perchlorate (10 and 100mgl−1), sodium arsenate (1 and 10mgl−1), and the mixture-sodium perchlorate+sodium arsenate (10+1mgl−1 and 100+10mgl−1 Na2HAsO4-high mixture) for 90d. Their uptake and accumulation by zebrafish was evaluated at 10, 30, 60, and 90d. In addition, depuration was examined at 1, 3, and 5d after cessation of the exposure. The uptake of either chemical was concentration-dependent, with significantly higher uptake at high concentrations at either exposure interval. In contrast, there was no significant difference in whole body residue between single chemicals and the corresponding mixture except for 100mgl−1 sodium arsenate at 90d. However, there was increasing accumulation over time at the high concentration of either chemical alone and their mixture, and this increasing trend was more pronounced in the single chemical exposures than in the mixture. At the concentrations tested in the current study, both chemicals reduced the uptake but enhanced the depuration of the other chemical from the zebrafish. This study represents the first examination of the interaction of two anions-perchlorate and arsenate with respect to toxicokinetics.

Constitutive Expression of a High‐Affinity Sulfate Transporter in Indian Mustard Affects Metal Tolerance and Accumulation

The Stylosanthes hamata SHST1 gene encodes a high-affinity sulfate transporter located in the plasma membrane. In this study the S. hamata SHST1 gene was constitutively expressed in Indian mustard [Brassica juncea (L.) Czern.] to investigate its importance for tolerance and accumulation of various oxyanions that may be transported by SHST1 and for cadmium, which is detoxified by sulfur-rich compounds. The transgenic SHST1 lines SHST1-12C and SHST1-4C were compared with wild-type Indian mustard for tolerance and accumulation of arsenate, chromate, tungstate, vanadate, and cadmium. As seedlings the SHST1 plants accumulated significantly more Cd and W, and somewhat more Cr and V. The SHST1 seedlings were less tolerant to Cd, Mo, and V compared to wild-type plants. Mature SHST1 plants were less tolerant than wild-type plants to Cd and Cr. SHST1 plants accumulated significantly more Cd, Cr, and W in their roots than wild-type plants. In their shoots they accumulated significantly more Cr and somewhat more V and W. Shoot Cd accumulation was significantly lower than in wild-type, and As levels were somewhat reduced. Compared to wild-type plants, sulfur accumulation was enhanced in roots of SHST1 plants but not in shoots. Together these results suggest that SHST1 can facilitate uptake of other oxyanions in addition to sulfate and that SHST1 mediates uptake in roots rather than root-to-shoot translocation. Since SHST1 overexpression led to enhanced accumulation of Cr, Cd, V, and W, this approach shows some potential for phytoremediation, especially if it could be combined with the expression of a gene that confers enhanced metal translocation or tolerance.

Effects of arsenate and phosphate on their accumulation by an arsenic-hyperaccumulator Pteris vittata L.

Arsenate and phosphate interactions are important for better understanding their uptake and accumulation by plant due to their similarities in chemical behaviors. The present study examined the effects of arsenate and phosphate on plant biomass and uptake of arsenate and phosphate by Chinese brake (Pteris vittata L.), a newly-discovered arsenic hyperaccumulator. The plants were grown for 20 weeks in a soil, which received the combinations of 670, 2670, or 5340 μmol kg−1 arsenate and 800, 1600, or 3200 μmol kg−1 phosphate, respectively. Interactions between arsenate and phosphate influenced their availability in the soil, and thus plant growth and uptake of arsenate and phosphate. At low and medium arsenate levels (670 and 2670 μmol kg−1), phosphate had slight effects on arsenate uptake by and growth of Chinese brake. However, phosphate substantially increased plant biomass and arsenate accumulation by alleviating arsenate phytotoxicity at high arsenate levels (5340 μmol kg−1). Moderate doses of arsenate increased plant phosphate uptake, but decreased phosphate concentrations at high doses because of its phytotoxicity. Based on our results, the minimum P/As molar ratios should be at least 1.2 in soil solution or 1.0 in fern fronds for the growth of Chinese brake. Our findings suggest that phosphate application may be an important strategy for efficient use of Chinese brake to phytoremediate arsenic contaminated soils. Further study is needed on the mechanisms of interactive effects of arsenate and phosphate on Chinese brake in hydroponic systems.

Mechanisms of arsenate tolerance in Cytisus striatus.

• Arsenate tolerance, uptake and arsenate-induced phytochelatin (PC) accumulation were compared at different phosphorus supply rates in two populations of the broom, Cytisus striatus , one from an arsenic-enriched gold mine and one from a nonmetalliferous site. • After 7d of exposure, arsenate tolerance was higher in the mine population. Arsenate uptake was phosphate-suppressible, and much lower in the mine plants. When compared at equal levels of stress, the mine plants and the nonmetallicolous plants exhibited similar arsenic accumulation, suggesting that reduced arsenate uptake is mainly responsible for superior tolerance. • Arsenate-induced PC accumulation occurred in both plant types. The γ-glutamylcysteine synthetase inhibitor, L-buthioninesulfoximine, caused arsenate hypersensitivity in both plant types, suggesting that PC-based arsenic sequestration is essential for both normal and enhanced arsenate tolerance. Mine plants produced longer PCs than the nonmetallicolous plants, possibly due to a differential temporal pattern of arsenate accumulation. • Our results are consistent with a similar mechanism underlying arsenate hypertolerance in C. striatus and grasses, that is reduced arsenate uptake through suppression of phosphate transporter activity.

Chromosomally encoded arsenical resistance of the moderately thermophilic acidophile Acidithiobacillus caldus.

Arsenical resistance is important to bioleaching microorganisms because these organisms release arsenic from minerals such as arsenopyrite during bioleaching. The acidophile Acidithiobacillus caldus KU was found to be resistant to the arsenical ions arsenate, arsenite, and antimony via an inducible, chromosomally encoded resistance mechanism. Because no apparent alteration of the toxic ions was observed, Acidithiobacillus (At.) caldus was tested to determine if it was resistant as a result of decreased accumulation of toxic ions. Reduced accumulation of arsenate and arsenite by induced At. caldus cells supported this hypothesis. It was also found that, with the addition of an energy source, induced At. caldus could transport arsenate and arsenite out of the cell against a concentration gradient. The lack of efflux in the absence of an added energy source and in the presence of inhibitors suggested that efflux was energy dependent. Induced At. caldus also expressed arsenate reductase activity, indicating that At. caldus has an arsenical resistance mechanism that is analogous to previously described systems from other Bacteria. Southern hybridization analysis showed that At. caldus and other gram-negative acidophiles carry an Escherichia coli arsB homologue on the chromosome.

ACCUMULATION AND REDUCTION OF ARSENATE BY THE FRESHWATER GREEN ALGA CHLORELLA SP. (CHLOROPHYTA).

Arsenate accumulation and reduction kinetics at both high and low phosphate concentrations were investigated in the green alga Chlorella sp, isolated from the arsenic-contaminated Upper Mystic Lake near Boston, MA. Growth rate, accumulated cellular arsenic, and release of As(III) were determined over a range of arsenate concentrations. Arsenate inhibited growth and reduced final cell yield at high phosphate concentration. However, growth rate, final cell yield, and cellular arsenic content were all enhanced by higher arsenate concentrations in cultures grown at a low concentration of phosphate. The traditional view that phosphate-limited cells are necessarily more sensitive to As(V) toxicity may not be correct. The reduction rates of As(V) by Chlorella sp. obtained in our laboratory were similar to net reduction rates measured in epilimnetic water from the Upper Mystic Lake, demonstrating the importance of phytoplankton in arsenic reduction in freshwater.

Edaphic factors affecting the toxicity and accumulation of arsenate in the earthwormLumbricus terrestris

AbstractThe toxicity and accumulation of arsenate was determined in the earthwormLumbricus terrestrisin soil from different layers of a forest profile. Toxicity increased fourfold between 2 and 10 d. Edaphic factors (pH, soil organic matter, and depth in soil profile) also affected toxicity with a three fold decrease in the concentration that causes 50% mortality with increasing depth in soil (from 0–70 mm to 500–700 mm). In a 4‐d exposure study, there was no evidence of arsenic bioconcentration in earthworm tissue, although bioaccumulation was occurring. There was a considerable difference in tissue residues between living and dead earthworms, with dead worms having higher concentrations. This difference was dependent on both soil arsenate concentration and on soil type. Over a wide range of soil arsenate concentrations, earthworm arsenic residues are homeostatically maintained in living worms, but this homeostasis breaks down during death. Alternatively, equilibration with soil residues may occur via accumulation after death. In long‐term accumulation studies in soils dosed with a sublethal arsenate concentration (40 μg/g dry weight), bioconcentration of arsenate did not occur until day 12, after which earthworm concentrations rose steadily above the soil concentration, with residues in worms three fold higher than soil concentrations by the termination of the study (23 d). This bioconcentration only occurred in depurated worms over the time period of the study. Initially, depurated worms had lower arsenic concentrations than undepurated until tissue concentrations were equivalent to the soil concentration. Once tissue concentration was greater than soil concentration, depurated worms had higher arsenic residues than undepurated.

EDAPHIC FACTORS AFFECTING THE TOXICITY AND ACCUMULATION OF ARSENATE IN THE EARTHWORM LUMBRICUS TERRESTRIS

The toxicity and accumulation of arsenate was determined in the earthworm Lumbricus terrestris in soil from different layers of a forest profile. Toxicity increased fourfold between 2 and 10 d. Edaphic factors (pH, soil organic matter, and depth in soil profile) also affected toxicity with a three fold decrease in the concentration that causes 50% mortality with increasing depth in soil (from 0–70 mm to 500–700 mm). In a 4-d exposure study, there was no evidence of arsenic bioconcentration in earthworm tissue, although bioaccumulation was occurring. There was a considerable difference in tissue residues between living and dead earthworms, with dead worms having higher concentrations. This difference was dependent on both soil arsenate concentration and on soil type. Over a wide range of soil arsenate concentrations, earthworm arsenic residues are homeostatically maintained in living worms, but this homeostasis breaks down during death. Alternatively, equilibration with soil residues may occur via accumulation after death. In long-term accumulation studies in soils dosed with a sublethal arsenate concentration (40 μg/g dry weight), bioconcentration of arsenate did not occur until day 12, after which earthworm concentrations rose steadily above the soil concentration, with residues in worms three fold higher than soil concentrations by the termination of the study (23 d). This bioconcentration only occurred in depurated worms over the time period of the study. Initially, depurated worms had lower arsenic concentrations than undepurated until tissue concentrations were equivalent to the soil concentration. Once tissue concentration was greater than soil concentration, depurated worms had higher arsenic residues than undepurated.

Accumulation of arsenic from acidic mine waters by ferruginous bacterial accretions (stromatolites)

In the acidic stream (pH 2.2–4) of the Carnoulès Pb-(Zn) mine, Gard, France, very high As contents (from 9 to 20%) can be accumulated as ferric arsenate and arsenate-sulphate precipitates in rapidly growing bacteria-made structures. The main bacterial forms are rod-shaped and sheathed, their sheath is made of Fe-As-rich material and is coated with ferric arsenate colloidal particles or may be partially included in authigenic crystals. Living forms of Thiobacillus-type bacteria have been recognized in the precipitates. The cyclic development of bacterial colonies alternating with sand deposition and erosive episodes results in the formation of As-rich ferruginous accretions. These laminated and dome-shaped bacterial constructions are similar to those of stromatolites. The extremely high contents of solute As in upstream flow (250 mg/1) are lowered by 2–3 order of magnitude downstream. Lead is also precipitated and concentrated in this Fe As-rich bacterial stromatolite (2500 ppm Pb). This accumulation and concentration of As and heavy metals via direct or induced microbial action limits pollution downflow. But seasonal storms could erode these Fe As Pb-rich deposits and drastically increase pollution. The accumulation of ferric arsenate by bacterial stromatolites suggests that possible microbial remediation strategies may be used in acid mine drainage environments.

Phosphorus Nutrition of Arsenate‐Tolerant and Nontolerant Phenotypes of Velvetgrass

AbstractVelvetgrass (Holcus lanatus L.), also known as Yorkshire fog grass, has evolved tolerance to high levels of arsenate, and this adaptation involves reduced accumulation of arsenate through the suppression of the high affinity phosphate‐arsenate uptake system. To determine the role of P nutrition in arsenate tolerance, inhibition kinetics of arsenate influx by phosphate were determined. The concentration of inhibitor required to reduce maximum influx (Vmax) by 50%, K1, of phosphate inhibition of arsenate influx was 0.02 mol m−3 in both tolerant and nontolerant clones. This was compared with the concentration where influx is 50% of maximum, a Km, for arsenate influx of 0.6 mol m−3 for tolerants and 0.025 mol m−3 for nontolerants and, therefore, phosphate was much more effective at inhibiting arsenate influx in tolerant genotypes. The high affinity phosphate uptake system is inducible under low plant phosphate status, this increasing plant phosphate status should increase tolerance by decreasing arsenate influx. Root extension in arsenate solutions of tolerant and nontolerant tillers grown under differing phosphate nutritional regimes showed that indeed, increased plant P status increased the tolerance to arsenate of both tolerant and nontolerant clones. That plant P status increased tolerance again argues that P nutrition has a critical role in arsenate tolerance. To determine if short term flux and solution culture studies were relevant to As and P accumulation in soils, soil and plant material from a range of As contaminated sites were analyzed. As predicted from the short‐term competition studies, P was accumulated preferentially to As in arsenate tolerant clones growing on mine spoil soils even when acid extractable arsenate in the soils was much greater than acid extractable phosphate. Though phosphate was much more efficient at competing with arsenate for uptake, plants growing on arsenate contaminated land still accumulated considerable amounts of As. Plants from the differing habitats showed large variation in plant phosphate status, pasture plants having much higher P levels than plants growing on the most contaminated mine spoil soils. The selectivity of the phosphate‐arsenate uptake system for phosphate compared with arsenate, coupled with the suppression of this uptake system enabled tolerant clones of the grass velvetgrass to grow on soils that were highly contaminated with arsenate and deficient in phosphate.

Arsenic accumulation in the shore crab Carcinus maenas: the influence of nutritional state, sex and exposure concentration

The accumulation of arsenate from seawater by the shore crab Carcinus maenas L. (collected from Odense Fjord, Denmark in 1991 and from Restronguet Creek, UK in 1991) was investigated in a series of laboratory experiments. A field study was also carried out to determine the effects of raised environmental arsenic concentrations on intra-organismal distribution and tissue concentrations. Studies on the influence of nutritional state and sex on accumulation of As(5) from seawater indicated that most of the arsenic taken up from seawater in laboratory experiments was retained in the gills and the midgut gland. Arsenic accumulation exhibited sex-dependent differences which were also evident in correlation analyses carried out between total lipid contents and total arsenic contents of midgut glands of individual crabs. Arsenic concentrations in the gonads of both sexes were strongly influenced by the nutritional state of the crabs. Elevated arsenic concentrations in seawater and food at an arsenic polluted site (Restronguet Creek) significantly influenced arsenic concentrations and distribution among the tissues of C. maenas. Arsenic concentrations and distribution patterns differed markedly from those crabs from an unpolluted site in Odense Fjord. The gills of the crabs from Restronguet Creek contained extremely high arsenic concentrations ranging from 179 to 483 μg As g-1 dry wt. These values were even higher than those measured in the gills of Odense crabs that had been exposed to 3 mgl-1 As(5) for 2 wk in the laboratory. Arsenic concentrations in the exoskeleton of Odense Fjord crabs were 15 times lower than those measured in exoskeletons of Restronguet Creek crabs. Approximately 69% of the total body burden of arsenic was located in muscle tissue of crabs from Odense Fjord, whereas the major pool of arsenic (46%) in Restronguet Creek crabs was located in the exoskeleton.

Arsenic efflux governed by the arsenic resistance determinant of Staphylococcus aureus plasmid pI258

The arsenic resistance operon of Staphylococcus aureus plasmid pI258 determined lowered net cellular uptake of 73As by an active efflux mechanism. Arsenite was exported from the cells; intracellular arsenate was first reduced to arsenite and then transported out of the cells. Resistant cells showed lower accumulation of 73As originating from both arsenate and arsenite. Active efflux from cells loaded with arsenite required the presence of the plasmid-determined arsB gene. Efflux of arsenic originating as arsenate required the presence of the arsC gene and occurred more rapidly with the addition of arsB. Inhibitor studies with S. aureus loaded with arsenite showed that arsenite efflux was energy dependent and appeared to be driven by the membrane potential. With cells loaded with 73AsO4(3-), a requirement for ATP for energy was observed, leading to the conclusion that ATP was required for arsenate reduction. When the staphylococcal arsenic resistance determinant was cloned into Escherichia coli, lowered accumulation of arsenate and arsenite and 73As efflux from cells loaded with arsenate were also found. Cloning of the E. coli plasmid R773 arsA gene (the determinant of the arsenite-dependent ATPase) in trans to the S. aureus gene arsB resulted in increased resistance to arsenite.

Uptake, accumulation and translocation of arsenate in arsenate‐tolerant and non‐tolerant Holcus lanatus L.

summaryUptake, translocation and accumulation of arsenate was determined in arsenate‐tolerant and non‐tolerant genotypes of Holcus lanatus L. Over a 6 h period of growth in 005 mol m−3arsenate, non‐tolerant genotypes accumulated arsenate to a much greater extent than tolerant plants. Tolerant plants transported a much greater proportion of As to their shoots compared with non‐tolerant plants. Phosphate at a concentration of either 0.05 or 0.5 mol m−3decreased arsenate uptake in both tolerant and non‐tolerant genotypes. When arsenate uptake was determined over 3 d at the same arsenate concentration, non‐tolerant plants grown in the presence of 0 and 0.05 mol m−3phosphate died, while those growing in 0.5 mol m−3phosphate survived and continued to take up arsenate. At all three phosphate levels tolerant plants survived. Over the 3 d period transport of arsenate to the shoots decreased. With increasing phosphate levels in solution transport of arsenate to the shoots increased in tolerant plants. The results from these experiments are discussed in terms of strategies for metal tolerance in this species.

Insight into the mechanism of accumulation of arsenate and phosphate in hydro lake sediments by measuring the rate of dissolution with ethylenediaminetetraacetic acid

The mechanism of accumulation of arsenic and phosphate on sediments in Lake Ohakuri in the North Island of New Zealand has been examined by comparing the rates of dissolution of the anions with the rates of dissolution of the matrix elements iron, aluminum, calcium, manganese, and silicon when samples were reacted with ethylenediaminetetraacetic acid (EDTA) at the natural pH of the sediments. Very close correlation was observed between the rates of dissolution of the anions and that of iron. This, together with the observation that iron not dissolved by EDTA was subsequently dissolved by Tamm's reagent, suggests that the most important mechanism of accumulation involves some interaction between hydrous iron oxides and the anions. From the constancy of the iron:arsenic and iron:phosphate ratios during dissolution it has been concluded that arsenic and phosphate are incorporated into these sediments by coprecipitation at the time of formation of the hydrous oxides rather than by adsorption on existing surfaces. 8 references, 4 figures, 5 tables.

Energy-dependent arsenate efflux: the mechanism of plasmid-mediated resistance.

Plasmid-mediated resistance to arsenate, arsenite, and antimony(III) is coordinately induced by arsenate, arsenite, antimony(III), and bismuth(III). Resistance to arsenate was recently shown [Silver, S., Budd, K., Leahy, K.M., Shaw, W.V., Hammond, D., Novick, R.P., Willsky, G.R., Malamy, M.H. & Rosenberg, H. (1981) J. Bacteriol. 146, 983-996] to be due to decreased accumulation of arsenate by the induced resistant cells. We report here that decreased net uptake results from accelerated efflux of arsenate by induced plasmid-containing cells of Staphylococcus aureus and Escherichia coli. The efflux system in S. aureus was inhibited by nigericin, monensin, and proton-mobilizing uncouplers; efflux was unaffected by valinomycin. The mechanism of arsenate efflux in S. aureus was apparently not by chemiosmotic coupling to the membrane electrical potential or pH gradient. The intracellular efflux system was inhibited by low pH and mercurials (reversible by mercaptoethanol). The efflux rate was relatively independent of external pH or phosphate level and showed a sigmoidal pattern of concentration dependence.

Effect of arsenate on inorganic phosphate transport in Escherichia coli.

The effect of arsenate on strains dependent on the two major inorganic phosphate (Pi) transport systems in Escherichia coli was examined in cells grown in 1 mM phosphate medium. The development of arsenate-resistant Pi uptake in a strain dependent upon the Pst (phosphate specific transport) system was examined. The growth rate of Pst-dependent cells in arsenate-containing medium was a function of the arsenate-to-Pi ratio. Growth in arsenate-containing medium was not due to detoxification of the arsenate. Kinetic studies revealed that cells grown with a 10-fold excess of arsenate to Pi have almost a twofold increase in capacity (Vmax) for Pi, but maintained the same affinity (Km). Pi accumulation in the Pst-dependent strain was still sensitive to changes in the arsenate-to-Pi ratio, and a Ki (arsenate) for Pi transport of 39 microM arsenate was determined. The Pst-dependent strain did not accumulate radioactive arsenate, and showed only a transient decrease in intracellular adenosine triphosphate levels after arsenate was added to the medium. The Pi transport-dependent strain ceased growth in arsenate-containing media. This strain accumulated 74As-arsenate, and intracellular adenosine triphosphate pools were almost completely depleted after the addition of arsenate to the medium. Arsenate accumulation required a metabolizable energy source and was inhibited by N-ethylmaleimide. Previously accumulated arsenate could exchange with arsenate or Pi in the medium.