Metal chelating properties of pyridine-2,6-bis(thiocarboxylic acid) produced by Pseudomonas spp. and the biological activities of the formed complexes

We evaluated the ability of pyridine-2,6-bis(thiocarboxylic acid) (pdtc) to form complexes with 19 metals and 3 metalloids. Pdtc formed complexes with 14 of the metals. Two of these metal:pdtc complexes, Co:(pdtc)₂ and Cu:pdtc, showed the ability to cycle between redox states, bringing to 4 the number of known redox-active pdtc complexes. A precipitant formed when pdtc was added to solutions of As, Cd, Hg, Mn, Pb, and Se. Additionally, 14 of 16 microbial strains tested were protected from Hg toxicity when pdtc was present. Pdtc also mediated protection from the toxic effects of Cd and Te, but for fewer strains. Pdtc by itself does not facilitate iron uptake, but increases the overall level of iron uptake of Pseudomonas stutzeri strain KC and P. putida DSM301. Both these pseudomonads could reduce amorphous Fe(III) oxyhydroxide in culture. In vitro reactions showed that copper and pdtc were required for this activity. This reaction may derive its reducing power from the hydrolysis of the thiocarboxyl groups of pdtc.     

Pyridine-2,6-Bis(Thiocarboxylic Acid) Produced by Pseudomonas stutzeri KC Reduces and Precipitates Selenium and Tellurium Oxyanions

The siderophore of Pseudomonas stutzeri KC, pyridine-2,6-bis(thiocarboxylic acid) (pdtc), is shown to detoxify selenium and tellurium oxyanions in bacterial cultures. A mechanism for pdtc's detoxification of tellurite and selenite is proposed. The mechanism is based upon determination using mass spectrometry and energy-dispersive X-ray spectrometry of the chemical structures of compounds formed during initial reactions of tellurite and selenite with pdtc. Selenite and tellurite are reduced by pdtc or its hydrolysis product H₂S, forming zero-valent pdtc selenides and pdtc tellurides that precipitate from solution. These insoluble compounds then hydrolyze, releasing nanometer-sized particles of elemental selenium or tellurium. Electron microscopy studies showed both extracellular precipitation and internal deposition of these metalloids by bacterial cells. The precipitates formed with synthetic pdtc were similar to those formed in pdtc-producing cultures of P. stutzeri KC. Culture filtrates of P. stutzeri KC containing pdtc were also active in removing selenite and precipitating elemental selenium and tellurium. The pdtc-producing wild-type strain KC conferred higher tolerance against selenite and tellurite toxicity than a pdtc-negative mutant strain, CTN1. These observations support the hypothesis that pdtc not only functions as a siderophore but also is involved in an initial line of defense against toxicity from various metals and metalloids.     

Antimicrobial Properties of Pyridine-2,6-Dithiocarboxylic Acid, a Metal Chelator Produced by Pseudomonas spp.

Pyridine-2,6-dithiocarboxylic acid (pdtc) is a metal chelator produced by Pseudomonas spp. It has been shown to be involved in the biodegradation of carbon tetrachloride; however, little is known about its biological function. In this study, we examined the antimicrobial properties of pdtc and the mechanism of its antibiotic activity. The growth of Pseudomonas stutzeri strain KC, a pdtc-producing strain, was significantly enhanced by 32 μM pdtc. All nonpseudomonads and two strains of P. stutzeri were sensitive to 16 to 32 μM pdtc. In general, fluorescent pseudomonads were resistant to all concentrations tested. In competition experiments, strain KC demonstrated antagonism toward Escherichia coli. This effect was partially alleviated by 100 μM FeCl₃. Less antagonism was observed in mutant derivatives of strain KC (CTN1 and KC657) which lack the ability to produce pdtc. A competitive advantage was restored to strain CTN1 by cosmid pT31, which restores pdtc production. pT31 also enhanced the pdtc resistance of all pdtc-sensitive strains, indicating that this plasmid contains elements responsible for resistance to pdtc. The antimicrobial effect of pdtc was reduced by the addition of Fe(III), Co(III), and Cu(II) and enhanced by Zn(II). Analyses by mass spectrometry determined that Cu(I):pdtc and Co(III):pdtc₂ form immediately under our experimental conditions. Our results suggest that pdtc is an antagonist and that metal sequestration is the primary mechanism of its antimicrobial activity. It is also possible that Zn(II), if present, may play a role in pdtc toxicity.     

Biogeochemical Elimination of Chromium (VI) from Contaminated Water

Ferrous iron [Fe(II)] reductively transforms heavy metals in contaminated groundwater, and the bacterial reduction of indigenous ferric iron [Fe(III)] to Fe(II) has been proposed as a means of establishing redox reactive barriers in the subsurface. The reduction of Fe(III) to Fe(II) can be accomplished by stimulation of indigenous dissimilatory metal-reducing bacteria (DMRB) or injection of DMRB into the subsurface. The microbially produced Fe(II) can chemically react with contaminants such as Cr(VI) to form insoluble Cr(III) precipitates. The DMRB Shewanella algae BrY reduced surface-associated Fe(III) to Fe(II), which in batch and column experiments chemically reduced highly soluble Cr(VI) to insoluble Cr(III). Once the chemical Cr(VI) reduction capacity of the Fe(II)/Fe(III) couple in the experimental systems was exhausted, the addition of S. algae BrY allowed for the repeated reduction of Fe(III) to Fe(II), which again reduced Cr(VI) to Cr(III). The research presented herein indicates that a biological process using DMRB allows the establishment of a biogeochemical cycle that facilitates chromium precipitation. Such a system could provide a means for establishing and maintaining remedial redox reactive zones in Fe(III)-bearing subsurface environments.     

Metal binding by pyridine-2,6-bis(monothiocarboxylic acid), a biochelator produced by Pseudomonas stutzeri and Pseudomonas putida

Pyridine-2,6-bis(monothiocarboxylic acid) (pdtc),a natural metal chelator produced by Pseudomonas stutzeri and Pseudomonas putidathat promotes the degradation of carbon tetrachloride, was synthesized and studiedby potentiometric and spectrophotometric techniques. The first two stepwise protonationconstants (pK) for successive proton addition to pdtc were found to be 5.48 and2.58. The third stepwise protonation constant was estimated to be 1.3. The stability (affinity)constants for iron(III), nickel(II), and cobalt(III) were determined by potentiometric orspectrophotometric titration. The results show that pdtc has strong affinity for Fe(III)and comparable affinities for various other metals. The stability constants (log K) are 33.93 for Co(pdtc)₂ ^1-; 33.36 for Fe(pdtc)₂ ^1-; and 33.28 for Ni(pdtc)₂ ^2-. These protonationconstants and high affinity constants show that over a physiological pH range theferric pdtc complex has one of the highest effective stability constants for ironbinding among known bacterial chelators.     

Bioreduction of Hexavalent Chromium from Soil Column Leachate by Pseudomonas stutzeri

Bioreduction of Cr(VI) to less toxic Cr(III) by chromate-reducing bacteria has offered an ecological and economical option for chromate detoxification. The present study reports isolation of chromate-resistant bacterial strain Cr8 from chromium slag, identified as Pseudomonas stutzeri, based on 16S rRNA gene sequencing and their potential use in Cr(VI) reduction. The reduced product associated with bacterial cell was characterized by scanning electron microscopy–energy-dispersive x-ray spectroscopy (SEM-EDS) and x-ray diffraction (XRD) analyses. At initial concentrations of 100 and 200 mg L^?1 Cr(VI), P. stutzeri Cr8 reduced Cr(VI) completely within 24 h, whereas it reduced almost 1000 mg L^?1 Cr(VI) at the end of 120 h. Further, soil column leaching experiments were performed and found that bacterial cells reduced Cr(VI) leachate at faster rate that almost disappeared at the end of 168 h. The leachate precipitates also revealed efficient chromate bioreduction. The remediation process utilizing P. stutzeri could be considered as a viable alternative to reduce Cr(VI) contamination, especially emanating from the overburden dumps of chromite ores and mine drainage.     

Effect of Zn(II) on the reduction and accumulation of Cr(VI) by <Emphasis Type="Italic">Arthrobacter</Emphasis> species

Natural habitats are often characterized by the coexistence of Zn and Cr. This study assessed the potential of two Gram-positive, Cr(VI)-reducing, aerobic bacterial strains belonging to Arthrobacter genera, which were isolated from basalt samples taken from the most polluted region of the Republic of Georgia, to remediate Cr(VI) in environments in the presence of Zn(II). Our batch experiments revealed that the addition of Zn(II) to the tested bacterial cells significantly enhanced the accumulation of Cr. According to electron spin resonance (ESR) measurements, the presence of Zn(II) ions did not change the nature of Cr(V) and Cr(III) complexes generated during the microbial reduction of Cr(VI). The efficiency of Cr(VI) reduction also remained unchanged after the addition of 50 mg/l of Zn(II) to the bacterial cells. However, at high concentrations of Zn(II) (higher than 200 mg/l), the transformation of Cr(VI) to Cr(V) and Cr(III) complexes decreases significantly. In addition, it was shown that the accumulation pattern of Zn in the tested bacterial species in the presence of 100 mg/l of Cr(VI) fits the Langmuir–Freundlich model well. The two tested bacterial strains exhibited different characteristics of Zn accumulation.     

Pyridine-2,6-bis(thiocarboxylic acid) produced by Pseudomonas stutzeri KC reduces and precipitates selenium and tellurium oxyanions

The siderophore of Pseudomonas stutzeri KC, pyridine-2,6-bis(thiocarboxylic acid) (pdtc), is shown to detoxify selenium and tellurium oxyanions in bacterial cultures. A mechanism for pdtc's detoxification of tellurite and selenite is proposed. The mechanism is based upon determination using mass spectrometry and energy-dispersive X-ray spectrometry of the chemical structures of compounds formed during initial reactions of tellurite and selenite with pdtc. Selenite and tellurite are reduced by pdtc or its hydrolysis product H(2)S, forming zero-valent pdtc selenides and pdtc tellurides that precipitate from solution. These insoluble compounds then hydrolyze, releasing nanometer-sized particles of elemental selenium or tellurium. Electron microscopy studies showed both extracellular precipitation and internal deposition of these metalloids by bacterial cells. The precipitates formed with synthetic pdtc were similar to those formed in pdtc-producing cultures of P. stutzeri KC. Culture filtrates of P. stutzeri KC containing pdtc were also active in removing selenite and precipitating elemental selenium and tellurium. The pdtc-producing wild-type strain KC conferred higher tolerance against selenite and tellurite toxicity than a pdtc-negative mutant strain, CTN1. These observations support the hypothesis that pdtc not only functions as a siderophore but also is involved in an initial line of defense against toxicity from various metals and metalloids.     

Biogeochemical Elimination of Chromium (VI) from Contaminated Water

Ferrous iron [Fe(II)] reductively transforms heavy metals in contaminated groundwater, and the bacterial reduction of indigenous ferric iron [Fe(III)] to Fe(II) has been proposed as a means of establishing redox reactive barriers in the subsurface. The reduction of Fe(III) to Fe(II) can be accomplished by stimulation of indigenous dissimilatory metal-reducing bacteria (DMRB) or injection of DMRB into the subsurface. The microbially produced Fe(II) can chemically react with contaminants such as Cr(VI) to form insoluble Cr(III) precipitates. The DMRB Shewanella algae BrY reduced surface-associated Fe(III) to Fe(II), which in batch and column experiments chemically reduced highly soluble Cr(VI) to insoluble Cr(III). Once the chemical Cr(VI) reduction capacity of the Fe(II)/Fe(III) couple in the experimental systems was exhausted, the addition of S. algae BrY allowed for the repeated reduction of Fe(III) to Fe(II), which again reduced Cr(VI) to Cr(III). The research presented herein indicates that a biological process using DMRB allows the establishment of a biogeochemical cycle that facilitates chromium precipitation. Such a system could provide a means for establishing and maintaining remedial redox reactive zones in Fe(III)-bearing subsurface environments.     

Antimicrobial properties of pyridine-2,6-dithiocarboxylic acid, a metal chelator produced by Pseudomonas spp

Pyridine-2,6-dithiocarboxylic acid (pdtc) is a metal chelator produced by Pseudomonas spp. It has been shown to be involved in the biodegradation of carbon tetrachloride; however, little is known about its biological function. In this study, we examined the antimicrobial properties of pdtc and the mechanism of its antibiotic activity. The growth of Pseudomonas stutzeri strain KC, a pdtc-producing strain, was significantly enhanced by 32 microM pdtc. All nonpseudomonads and two strains of P. stutzeri were sensitive to 16 to 32 microM pdtc. In general, fluorescent pseudomonads were resistant to all concentrations tested. In competition experiments, strain KC demonstrated antagonism toward Escherichia coli. This effect was partially alleviated by 100 microM FeCl3. Less antagonism was observed in mutant derivatives of strain KC (CTN1 and KC657) which lack the ability to produce pdtc. A competitive advantage was restored to strain CTN1 by cosmid pT31, which restores pdtc production. pT31 also enhanced the pdtc resistance of all pdtc-sensitive strains, indicating that this plasmid contains elements responsible for resistance to pdtc. The antimicrobial effect of pdtc was reduced by the addition of Fe(III), Co(III), and Cu(II) and enhanced by Zn(II). Analyses by mass spectrometry determined that Cu(I):pdtc and Co(III):pdtc2 form immediately under our experimental conditions. Our results suggest that pdtc is an antagonist and that metal sequestration is the primary mechanism of its antimicrobial activity. It is also possible that Zn(II), if present, may play a role in pdtc toxicity.     

Effect of organic Cr(III) complexes on chromium speciation

Abstract

Chromium speciation in the presence of organic chromium(III) complexes was investigated using solid-phase extraction. The adsorptions of Cr(VI) and Cr(III) on alumina and pumice powder were studied. Maximum sorption of Cr(VI) was obtained by alumina (90.22%), while Cr(III) was highly adsorbed onto pumice powder (86.65%). This result shows that pumice may be a new and promising adsorbent for Cr(III). The experimental equilibrium data for Cr(VI) adsorption onto alumina and Cr(III) sorption onto pumice were analysed using Langmuir and Freundlich isotherms. The separation and adsorption of Cr(VI), Cr(III) and five organic chromium(III) complexes onto pumice and alumina at different pH values were evaluated. Ethylenediaminetetraacetate (EDTA), oxalate, citrate, glycine, alanine and 8-hydroxyqinoline were used as ligands. Sorption of alanine and ethylenediaminetetraacetate complexes was higher onto alumina than pumice at pH>3. The enhancement of adsorption of chromium(III) complexes onto pumice was achieved by surface modification of pumice using a surfactant, namely hexadecyltrimethylammoniumbromür (HDTMA). The presence of surfactant enhanced the adsorption of Cr(III) citrate, oxalate, glycine and 8-hydroxyquinoline complexes onto pumice. However, the adsorption of EDTA and alanine complexes decreased, with ratio of 13.40% and 4.00% respectively. Here we demonstrate that chromium speciation methods depending on adsorption onto various adsorbents including alumina may lead erroneous results. Analytical measurements were performed by flame AAS, data were obtained by standard addition method.     

METAL‐INDUCED REACTIVE OXYGEN SPECIES PRODUCTION IN CHLAMYDOMONAS REINHARDTII (CHLOROPHYCEAE)1

Toxic effects of metals appear to be partly related to the production of reactive oxygen species (ROS), which can cause oxidative damage to cells. The ability of several redox active metals [Fe(III), Cu(II), Ag(I), Cr(III), Cr(VI)], nonredox active metals [Pb(II), Cd(II), Zn(II)], and the metalloid As(III) and As(V) to produce ROS at environmentally relevant metal concentrations was assessed. Cells of the freshwater alga Chlamydomonas reinhardtii P. A. Dang. were exposed to various metal concentrations for 2.5 h. Intracellular ROS accumulation was detected using an oxidation‐sensitive reporter dye, 5‐(and‐6)‐carboxy‐2′,7′‐dihydrodifluorofluorescein diacetate (H₂DFFDA), and changes in the fluorescence signal were quantified by flow cytometry (FCM). In almost all cases, low concentrations of both redox and nonredox active metals enhanced intracellular ROS levels. The hierarchy of maximal ROS induction indicated by the increased number of stained cells compared to the control sample was as follows: Pb(II) > Fe(III) > Cd(II) > Ag(I) > Cu(II) > As(V) > Cr(VI) > Zn(II). As(III) and Cr(III) had no detectable effect. The effective free metal ion concentrations ranged from 10^?6 to 10^?9 M, except in the case of Fe(III), which was effective at 10^?18 M. These metal concentrations did not affect algal photosynthesis. Therefore, a slightly enhanced ROS production is a general and early response to elevated, environmentally relevant metal concentrations.     

Role of extracellular polymeric substance (EPS) in toxicity response of soil bacteria Bacillus sp. S3 to multiple heavy metals

Heavy metal resistant bacteria are of great interest because of their potential use in bioremediation. Understanding the survival and adaptive strategies of these bacteria under heavy metal stress is important for better utilization of these bacteria in remediation. The objective of this study was to investigate the role of bacterial extracellular polymeric substance (EPS) in detoxifying against different heavy metals in Bacillus sp. S3, a new hyper antimony-oxidizing bacterium previously isolated from contaminated mine soils. The results showed that Bacillus sp. S3 is a multi-metal resistant bacterial strain, especially to Sb(III), Cu(II) and Cr(VI). Toxic Cd(II), Cr(VI) and Cu(II) could stimulate the secretion of EPS in Bacillus sp. S3, significantly enhancing the adsorption and detoxification capacity of heavy metals. Both Fourier transform infrared spectroscopy (FTIR) and three-dimensional excitation–emission matrix (3D-EEM) analysis further confirmed that proteins were the main compounds of EPS for metal binding. In contrast, the EPS production was not induced under Sb(III) stress. Furthermore, the TEM–EDX micrograph showed that Bacillus sp. S3 strain preferentially transported the Sb(III) to the inside of the cell rather than adsorbed it on the extracellular surface, indicating intracellular detoxification rather than extracellular EPS precipitation played an important role in microbial resistance towards Sb(III). Together, our study suggests that the toxicity response of EPS to heavy metals is associated with difference in EPS properties, metal types and corresponding environmental conditions, which is likely to contribute to microbial-mediated remediation.

     

Decomposition of Cr(V)-diols to Cr(III) Complexes by <Emphasis Type="Italic">Arthrobacter oxydans</Emphasis>

We demonstrated previously that Cr(VI) is readily reduced to oxoCr(V)-diols at the surface of Arthrobacter oxydans—a Gram-positive aerobic bacteria isolated from Columbia basalt rocks originated from a highly contaminated site in the USA. Here, we report an electron spin resonance (ESR) study of Cr(III) hydroxide formation from Cr(V)-diols by this bacterial strain as cells were exposed to 35, 200, and 400 mg/L of Cr(VI) under aerobic conditions as a batch culture and as lyophilized cells. The time-dependent ESR measurements show that the half-time of Cr(III) formation is almost equal to that of Cr(V) decomposition, which is in the range of 3–6 days for all cases. This rate is at least 300 times slower than that of Cr(V) formation. Additionally, atomic absorption spectrometry was also employed to examine the time course of total chromium in bacterial cells. This is the first time the kinetics of Cr(III) complexes formation in bacteria is evaluated.     

Reduction of chromium (VI) by reference bacterial strains

Collection bacterial strains were found to be capable of chromium (VI) reduction although they had not been in contact with chromium compounds before. Strains capable of nitrate respiration could use bichromate ions as a terminal electron acceptor in the absence of competing acceptors. Cr(VI) was reduced to Cr(III) when bichromate was added to the cultural broth whose redox potential reached -140 mV.     

Oxygen radical-mediated DNA damage by redox-active Cr(III) complexes.

The mechanism of DNA damage induced by Cr(III) complexes is currently unknown even though it is considered to be the ultimate biologically active oxidation state of chromium. In this study, we have employed the Salmonella reversion assay to identify mutagenic Cr(III) complexes. Cyclic voltammetry was used to differentiate the redox kinetics between mutagenic and selected nonmutagenic Cr(III) species. Plasmid relaxation of supercoiled DNA was employed to show in vitro interactions with plasmid DNA and correlate the interactions with the electrochemical behavior and biological activity. The results of this study demonstrate that the mutagenic Cr(III) complexes identified in the Salmonella reversion assay display characteristics of reversibility and positive shifts of the Cr(III)/Cr(II) redox couple consistent with the ability of these Cr(III) complexes to serve as cyclical electron donors in a Fenton-like reaction. These same mutagenic complexes display an ability to relax supercoiled DNA in vitro, presumably by the induction of single-strand breaks. Nonmutagenic complexes were selected to test different ligands to determine how the ligand directs the activity of Cr(III) complexes. All nonmutagenic complexes tested thus far have shown classical irreversibility, more negative reduction potentials, and an inability to relax supercoiled plasmid DNA. These results suggest that the mechanism by which chromium complexes potentiate mutagenesis involves an oxygen radical as an active intermediate. These data also demonstrate the effect of associated ligands with regard to the ability of a metal to generate an active redox center.     

Tolerance of a ruminant ciliate <Emphasis Type="Italic">Entodinium caudatum</Emphasis> against mercury,copper and chromium

The effects of three heavy metal cations, mercury (II), copper (II), and chromium (VI), on the growth of the rumen ciliate Entodinium caudatum in vitro culture was studied. The E. caudatum culture was challenged by HgCl₂, CuCl₂, and K₂Cr₂O₇ for a period of 4 days. The tested concentrations of mercury (II) and copper (II) were 1, 5, 10, 20, 50 mg/L and 2, 10, 20, 40 mg/L for chromium (VI) at single dose with either untreated or inhibited bacterial co-culture population. Effective metal concentrations required to reduce ciliate growth by 50% (EC₅₀) for mercury (II), copper (II), and chromium (VI) either with untreated or inhibited bacterial co-culture population after 24 h of metal application were 24, 20, and 21 or 15, 20, and 19 mg/L, respectively. After 4 days of metal application, corresponding EC₅₀ values for mercury (II), copper (II), and chromium (VI) were 16, 20, and 17 (with untreated bacterial population) or not determinable, 20, and 15 mg/L, respectively (with inhibited bacterial population). Increased sensitivity of E. caudatum to tested heavy metals with inhibited bacterial co-culture population indicate that the ciliate resistance to the heavy metal tested depends on detoxification abilities of rumen bacterial population.     

Addition of DNA to Cr(VI) and cytochrome b5 containing proteoliposomes leads to generation of DNA strand breaks and Cr(III) complexes

Chromium (Cr) is a cytotoxic metal that can be associated with a variety of types of DNA damage, including Cr-DNA adducts and strand breaks. Prior studies with purified human cytochrome b(5) and NADPH:P450 reductase in reconstituted proteoliposomes (PLs) demonstrated rapid reduction of Cr(VI) (hexavalent chromium, as CrO(4)(2-), and the generation of Cr(V), superoxide (O(2)(*-)), and hydroxyl radical (HO(*)). Studies reported here examined the potential for the species produced by this system to interact with DNA. Strand breaks of purified plasmid DNA increased over time aerobically, but were not observed in the absence of O(2). Cr(V) is formed under both conditions, so the breaks are not mediated directly by Cr(V). The aerobic strand breaks were significantly prevented by catalase and EtOH, but not by the metal chelator diethylenetriaminepentaacetic acid (DTPA), suggesting that they are largely due to HO(*) from Cr-mediated redox cycling. EPR was used to assess the formation of Cr-DNA complexes. Following a 10-min incubation of PLs, CrO(4)(2-), and plasmid DNA, intense EPR signals at g=5.7 and g=5.0 were observed. These signals are attributed to specific Cr(III) complexes with large zero field splitting (ZFS). Without DNA, the signals in the g=5 region were weak. The large ZFS signals were not seen, when Cr(III)Cl(3) was incubated with DNA, suggesting that the Cr(III)-DNA interactions are different when generated by the PLs. After 24 h, a broad signal at g=2 is attributed to Cr(III) complexes with a small ZFS. This g=2 signal was observed without DNA, but it was different from that seen with plasmid. It is concluded that EPR can detect specific Cr(III) complexes that depend on the presence of plasmid DNA and the manner in which the Cr(III) is formed.     

Fe(III), Cr(VI), and Fe(III) mediated Cr(VI) reduction in alkaline media using a <Emphasis Type="Italic">Halomonas</Emphasis> isolate from Soap Lake,Washington

Hexavalent chromium is one of the most widely distributed environmental contaminants. Given the carcinogenic and mutagenic consequences of Cr(VI) exposure, the release of Cr(VI) into the environment has long been a major concern. While many reports of microbial Cr(VI) reduction are in circulation, very few have demonstrated Cr(VI) reduction under alkaline conditions. Since Cr(VI) exhibits higher mobility in alkaline soils relative to pH neutral soils, and since Cr contamination of alkaline soils is associated with a number of industrial activities, microbial Cr(VI) reduction under alkaline conditions requires attention. Soda lakes are the most stable alkaline environments on earth, and contain a wide diversity of alkaliphilic organisms. In this study, a bacterial isolate belonging to the Halomonas genus was obtained from Soap Lake, a chemically stratified alkaline lake located in central Washington State. The ability of this isolate to reduce Cr(VI) and Fe(III) was assessed under alkaline (pH = 9), anoxic, non-growth conditions with acetate as an electron donor. Metal reduction rates were quantified using Monod kinetics. In addition, Cr(VI) reduction experiments were carried out in the presence of Fe(III) to evaluate the possible enhancement of Cr(VI) reduction rates through electron shuttling mechanisms. While Fe(III) reduction rates were slow compared to previously reported rates, Cr(VI) reduction rates fell within range of previously reported rates.     

The determination of the stability constant for the iron(II) complex of the biochelator pyridine-2,6-bis(monothiocarboxylic acid)

Pyridine-2,6-bis(monothiocarboxylic acid), also known as pyridine-2,6-dithiocarboxylic acid (pdtc), is a unique and powerful metal chelator produced by Pseudomonas stutzeri and Pseudomonas putida. The actual physiological roles of pdtc in these pseudomonads are not known with certainty, though it is likely that the compound acts as a siderophore, an antibiotic, or both. The stability constant of Fe^III(pdtc)₂ ^2- was determined in previous work to be 10^33.36. Here we determined that the stability constant of Fe^II(pdtc)₂ ^2- is 10¹². We determined this stability constant through potentiometric and spectrophotometric measurements of a ligand-ligand competition study using 2,6-pyridine dicarboxylic acid as the competitor for iron. Comparing the stability constant for Fe^II(pdtc)₂ ^2- to the constant for Fe^III(pdtc)₂ ^2- shows that the stability constant of Fe^II(pdtc)₂ ^2- is approximately 21 orders of magnitude smaller. This represents a very significant decrease in the binding strength of pdtc toward iron. Thus, if the host cell produces pdtc as a siderophore for sequestering Fe(III), it is likely that a second metabolite or a membrane protein of the host cell is used for reduction of the chelated iron at or near the cell membrane in order to facilitate its release from pdtc for cellular use.