Reducing Bacterium Shewanella Putrefaciens Research Articles

Effects of gamma(γ)-irradiation on the physicochemical properties and bioavailability of iron oxyhydroxides coprecipitated with varying concentrations of Na-alginate

This study investigated the impacts of γ-radiation at a final dose of 25 kGy on the physicochemical properties of a range of wet iron oxyhydroxides (2-line ferrihydrite, lepidocrocite, and goethite) synthesized in the presence of varying concentrations of the polysaccharide Na-alginate (starting solutions containing C/Fe ratios of 0, 0.5, 1.0, and 1.5). The degree of impact to the minerals was examined by chemical extractions, and various analytical techniques including XRD, FTIR-ATR, Mössbauer spectroscopy, and N2 adsorption-desorption isotherms, as well as by assessing their bioavailability towards the model Fe(III) reducing bacteria Shewanella putrefaciens CN32. Across all the coprecipitates studied, it was found that γ-irradiation led to a substantial dissolution of Fe and the concomitant release of Fe(II) and alginate into solution. Despite this observation, the bulk mineralogy and crystallinity of the studied iron oxyhydroxides, as determined by XRD and FTIR-ATR, did not appear to change. However, analyses via Mössbauer spectroscopy (77 and 5 K) revealed that the crystallinity of the 2-line ferrihydrites increased post-irradiation. Moreover, among the minerals studied, the specific surface area and porosity decreased for only the post-irradiated 2-line ferrihydrite coprecipitates with a C/Fe ratio of 0.5 or 1.0. The bioreduction rates of the studied minerals and their irradiated counterparts did not significantly differ, whereas the extent of bioreduction of post-irradiated 2-line ferrihydrite coprecipitates (C/Fe 0.5, 1.0, and 1.5) exhibited a significant increase of up to 28%. In contrast, the extent of Fe reduction for select post-irradiated lepidocrocite (C/Fe 0.5) and goethite (C/Fe 1.5) coprecipitates was slightly higher than determined for their corresponding native controls. The observed differences in bioavailability between the native and irradiated coprecipitates were attributed to irradiation induced alteration of particle aggregation and coagulation as determined by particle size analyses and visual observations. In summary, the findings suggest that even a low total dose of 25 kGy, γ-radiation can lead to significant physicochemical changes in coprecipitates with relatively low organic matter content. Therefore, future research designed to investigate organic matter-Fe composite systems in natural samples should be cognizant of the potential effects of sterilization through γ-irradiation. These effects could potentially result in inaccurate over- or under- estimations of the bioavailability of Fe or organic matter, as well as the possible adsorption capacity of organic matter-Fe coprecipitates for contaminants.

Biofilm Development on Carbon Steel by Iron Reducing Bacterium Shewanella putrefaciens and Their Role in Corrosion

Microscopic, electrochemical and surface characterization techniques were used to investigate the effects of iron reducing bacteria (IRB) biofilm on carbon steel corrosion for 72 and 168 h under batch conditions. The organic nutrient availability for the bacteria was varied to evaluate biofilms formed under nutritionally rich, as compared to nutritionally deficient, conditions. Focused ion beam-scanning electron microscopy (FIB-SEM) was used to investigate the effect of subsurface biofilm structures on the corrosion characteristics of carbon steel. Hydrated biofilms produced by IRB were observed under environmental scanning electron microscope (ESEM) with minimal surface preparation, and the elemental composition of the biofilms was investigated using energy dispersive spectroscopy (EDX). Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) was used to provide information on the organic and inorganic chemical makeup of the biofilms. Electrochemical techniques employed for assessing corrosion, by open circuit potential, linear polarization and potentiodynamic polarization tests indicated no significant difference in the corrosion resistance for carbon steel in IRB-inoculated, compared to the abiotic solutions of common Postgate C after 72 and 168 h. However, the steel was found to be more susceptible to corrosion when the yeast extract was removed from the biotic environment for the 168 h test. In the absence of yeast nutrient, it is postulated that IRB received energy by transforming the protective film of Fe3+ into more soluble Fe2+ products.

Lignin-enhanced reduction of structural Fe(III) in nontronite: Dual roles of lignin as electron shuttle and donor

Lignin is a major component of plant-derived soil organic matter (SOM) in soils and sediments. Fe-bearing clay minerals are widely distributed in these environments and often co-exist with lignin. While previous studies have reported the electron shuttling and donating roles of certain redox-active SOM in the dissimilatory reduction of structural Fe(III) in Fe-bearing clay minerals, the role of lignin in this process remains unknown. Here we studied this role by incubating an Fe-rich smectite (nontronite NAu-2) with two types of lignin (soluble and insoluble) in the absence and presence of an Fe(III)-reducing bacterium Shewanella putrefaciens CN32 under anaerobic condition. Lactate was added in some experiments as an extra electron donor. The results demonstrated that both soluble and insoluble lignins abiotically reduced structural Fe(III) in NAu-2. The reduction extent was proportional to lignin concentration. After abiotic reaction, lignin served as either electron shuttle or electron donor in the presence of CN32: (1) When lactate was present, lignin served as an electron shuttle to enhance the rate of Fe(III) reduction; (2) When lactate was absent, lignin served as an electron donor for Fe(III) reduction. Although the ultimate biotic Fe(III) reduction extents were similar in the presence of either soluble or insoluble lignin, the reduction rates with soluble lignin were higher than those with insoluble lignin, likely owing to their different electron transfer mechanisms. After interaction with NAu-2 and/or CN32, soluble lignin structure largely remained intact, but with some decreases of humic/fulvic acid-like and protein-like compounds, aromatic functional groups (e.g., CH, CO, COOH), and aliphatic/aromatic compounds. An increase of semiquinone-like organic radicals was observed after lignin interaction with NAu-2. These chemical changes of lignin were likely coupled with the reduction of structural Fe(III) in nontronite. Upon reduction, the nontronite did not display much dissolution and mineral transformation. The findings of this study provide insights into the role of lignin in promoting mineral-microbe interactions and have significant implications for coupled Fe and C biogeochemical cycles in soils and sediments.

Abundance of Fe(III) during cultivation affects the microbiologically influenced corrosion (MIC) behaviour of iron reducing bacteria Shewanella putrefaciens

The effect of the presence of Fe(III) during the cultivation on the electrochemical activity and corrosion behaviour of dissimilatory iron reducing bacteria Shewanella putrefaciens was studied by means of ex situ and in situ X-ray absorption near-edge spectroscopy (XANES). Stainless steel AISI 304 and thin iron films were studied as substrates. XANES analysis indicated an accelerated iron dissolution and growth of an oxide/hydroxide film for the culture grown with Fe(III) in comparison to the culture grown in absence of Fe(III). Electrochemical analysis indicated that the biofilm resulted in acceleration of the general corrosion but provides protection against local corrosion.

Effect of Indigenous Microbial Consortium on Bioleaching of Arsenic from Contaminated Soil by Shewanella putrefaciens

The effects of indigenous microbial consortium on removal of As from As-contaminated soil using an Fe(III)-reducing bacterium Shewanella putrefaciens were investigated under circumneutral pH condition. Sequential extraction of As revealed that more than 30% of As was associated with Fe(III)-(oxy)hydroxides in the soil. Bioleaching experiments were conducted anaerobically with a supply of lactate as a carbon source. The highest As removal efficiency (57.5%) was obtained when S. putrefaciens and indigenous bacterial consortium coexisted in the soil. S. putrefaciens and indigenous bacteria solely removed 30.1% and 16.4% of As from the soil, respectively. The combination of S. putrefaciens and indigenous bacteria led to a higher amount of labile As after microbial dissolution of Fe(III)-(oxy)hydroxides. After microbial treatment, soil quality represented by pH and organic content appeared to be preserved. The results indicated that the ecological and physiological understanding of the indigenous microbiome might be important for the efficient application of bioleaching technology to remove As from contaminated soils.

Trained to corrode: Cultivation in the presence of Fe(III) increases the electrochemical activity of iron reducing bacteria – An in situ electrochemical XANES study

This paper reports results from in situ electrochemical X-ray absorption near-edge spectroscopy (XANES) studies of the corrosion processes on model thin iron films in the presence of iron reducing bacteria Shewanella putrefaciens. Here we investigate the electrochemical activity of two cultures grown in the presence and absence of Fe(III) citrate in the culture medium. The XANES spectra and the OCP data of the Fe sample incubated with the culture grown in absence of Fe(III) did not show any significant changes during twenty hours of monitoring. In the case of the culture grown in Fe(III) containing medium, an accelerated dissolution of the iron film was observed together with the formation of a mixed Fe(II)-Fe(III) hydroxide surface layer. The open circuit potential (OCP) steadily approached the free corrosion potential of iron in neutral chloride containing electrolytes, indicating a continuous dissolution process without passivation.

Microbial mediated sedimentary phosphorus mobilization in emerging and eroding wetlands of coastal Louisiana

The interactions between the microbial reduction of Fe (III) oxides and sediment geochemistry are poorly understood and mostly unknown for the Louisiana deltaic plain. This study evaluates the potential of P mobilization for this region during bacterially mediated redox reactions. Samples were collected from two wetland habitats (forested wetland ridge, and marsh) characterized by variations in vegetation structure and elevation in the currently prograding Wax Lake Delta (WLD) and two habitats (wetland marsh, and benthic channel) in degrading Barataria Bay in Lake Cataouatche (BLC). Our results show that PO43− mobilization from WLD and BLC habitats were negligible under aerobic condition. Under anaerobic condition, there is a potential for significant release of PO43− from sediment and wetland soils. PO43− release in sediments spiked with Fe reducing bacteria Shewanella putrefaciens (Sp-CN32) were significantly higher in all cases with respect to a control treatment. In Wax Lake delta, PO43− release from sediment spiked with Sp-CN32 increased significantly from 0.064 ± 0.001 to 1.460 ± 0.005 μmol g−1 in the ridge and from 0.079 ± 0.007 to 2.407 ± 0.001 μmol g−1 in the marsh substrates. In Barataria bay, PO43− release increased from 0.103 ± 0.006 μmol g−1 to 0.601 ± 0.008 μmol g−1 in the channel and 0.050 ± 0.000 to 0.618 ± 0.026 μmol g−1 in marsh substrates. The PO43− release from sediment slurries spiked with Sp-CN32 was higher in the WLD habitats (marsh 30-fold, ridge 22-fold) compared to the BLC habitats (marsh 12-fold, channel 6-fold). The increase in PO43− release was significantly correlated with the Fe bound PO43− in sediments from different habitats but not with their organic matter content. This study contributes to our understanding of the release mechanism of PO43− during bacterial mediated redox reaction in wetland soils undergoing pulsing sediment deposition and loss.

Biological redox cycling of iron in nontronite and its potential application in nitrate removal.

Biological redox cycling of structural Fe in phyllosilicates is an important but poorly understood process. The objective of this research was to study microbially mediated redox cycles of Fe in nontronite (NAu-2). During the reduction phase, structural Fe(III) in NAu-2 served as electron acceptor, lactate as electron donor, AQDS as electron shuttle, and dissimilatory Fe(III)-reducing bacterium Shewanella putrefaciens CN32 as mediator in bicarbonate- and PIPES-buffered media. During the oxidation phase, biogenic Fe(II) served as electron donor and nitrate as electron acceptor. Nitrate-dependent Fe(II)-oxidizing bacterium Pseudogulbenkiania sp. strain 2002 was added as mediator in the same media. For all three cycles, structural Fe in NAu-2 was able to reversibly undergo three redox cycles without significant dissolution. Fe(II) in bioreduced samples occurred in two distinct environments, at edges and in the interior of the NAu-2 structure. Nitrate reduction to nitrogen gas was coupled with oxidation of edge-Fe(II) and part of interior-Fe(II) under both buffer conditions, and its extent and rate did not change with Fe redox cycles. These results suggest that biological redox cycling of structural Fe in phyllosilicates is a reversible process and has important implications for biogeochemical cycles of carbon, nitrogen, and other nutrients in natural environments.

Competitive reduction of nitrate and iron oxides by Shewanella putrefaciens 200 under anoxic conditions

While the competitive reduction of nitrate and iron oxides has been documented previously, the detailed kinetics and the influence of iron oxides’ properties remain unclear. In this study, the kinetics for the microbial reduction of nitrate and iron oxides by a dissimilatory iron reducing bacterium Shewanella putrefaciens 200 (SP200) were investigated in artificial groundwater medium containing nitrate and various synthetic iron oxides. All iron oxides exhibited the inhibition of nitrate reduction by SP200, and the reduction of those iron oxides by SP200 was simultaneously inhibited, as well. The effects of crystalline α-Fe2O3 indicated that the extent of nitrate reduction inhibition decreased gradually in the presence of increasing crystalline hematite. The competitive effects were also confirmed by the cyclic voltammetry analysis and spectral analysis of cytochromes in whole cells of SP200. The competition between nitrate and ferric reduction might be ascribed to (1) the competition of Fe (III) and nitrate as coexisting electron acceptors for iron-reducing bacteria; (2) the slow reaction of cell-sorbed Fe(II) with nitrate compared with the direct reduction of nitrate by SP200. This study would provide useful information concerning the redox cycles of N in various iron-bearing environments.

Dissimilatory Reduction and Transformation of Ferrihydrite-Humic Acid Coprecipitates

Organic matter (OM) is present in most terrestrial environments and is often found coprecipitated with ferrihydrite (Fh). Sorption or coprecipitation of OM with Fe oxides has been proposed to be an important mechanism for long-term C preservation. However, little is known about the impact of coprecipitated OM on reductive dissolution and transformation of Fe(III) (oxyhydr)oxides. Thus, we study the effect of humic acid (HA) coprecipitation on Fh reduction and secondary mineral formation by the dissimilatory Fe(III)-reducing bacterium Shewanella putrefaciens strain CN32. Despite similar crystal structure for all coprecipitates investigated, resembling 2-line Fh, the presence of coprecipitated HA resulted in lower specific surface areas. In terms of reactivity, coprecipitated HA resulted in slower Fh bioreduction rates at low C/Fe ratios (i.e., C/Fe ≤ 0.8), while high C/Fe ratios (i.e., C/Fe ≥ 1.8) enhanced the extent of bioreduction compared to pure Fh. The coprecipitated HA also altered the secondary Fe mineralization pathway by inhibiting goethite formation, reducing the amount of magnetite formation, and increasing the formation of a green rust-like phase. This study indicates that coprecipitated OM may influence the rates, pathway, and mineralogy of biogeochemical Fe cycling and anaerobic Fe respiration within soils.

Effects of Electron Transfer Mediators on the Bioreduction of Lepidocrocite (γ-FeOOH) by Shewanella putrefaciens CN32

Electron transfer mediators (ETMs) such as low-molecular-mass quinones (e.g., juglone and lawsone) and humic substances are believed to play a role in many redox reactions involved in contaminant transformations and the biogeochemical cycling of many redox-active elements (e.g., Fe and Mn) in aquatic and terrestrial environments. This study examines the effects of a series of compounds representing major classes of natural and synthetic organic ETMs, including low-molecular-mass quinones, humic substances, phenazines, phenoxazines, phenothiazines, and indigo derivatives, on the bioreduction of lepidocrocite (gamma-FeOOH) by the dissimilatory Fe(III)-reducing bacterium Shewanella putrefaciens CN32. Although S. putrefaciens CN32 was able to reduce lepidocrocite in the absence of exogenous ETMs, the addition of exogenous ETMs enhanced the bioreduction of lepidocrocite. In general, the rate of Fe(II) production correlated well with the reduction potentials of the ETMs. The addition of humic acids or unfractionated natural organic matter at concentrations of 10 mg organic CL(-1) resulted in, at best, a minimal enhancement of lepidocrocite bioreduction. This observation suggests that electron shuttling by humic substances is not likely to play a major role in Fe(lll) bioreduction in oligotrophic environments such as subsurface sediments with low organic C contents.

Confounding Impacts of Iron Reduction on Arsenic Retention

A transition from oxidizing to reducing conditions has long been implicated to increase aqueous As concentrations, for which reductive dissolution of iron (hydr)oxides is commonly implicated as the primary culprit. Confounding our understanding of processes controlling As retention, however, is that reductive transformation of ferrihydrite has recently been shown to promote As retention rather than release. To resolve the role iron phases have in regulating arsenic concentrations, here we examine As desorption from ferrihydrite-coated sands presorbed with As(III); experiments were performed at circumneutral pH under Fe-reducing conditions with the dissimilatory iron reducing bacterium Shewanella putrefaciens strain CN-32 over extended time periods. We reveal that with the initial phase of iron reduction, ferrihydrite undergoes transformation to secondary phases and increases As(III) retention (relative to abiotic controls). However, with increased reaction time, cessation of the phase transitions and ensuing reductive dissolution result in prolonged release of As(III) to the aqueous phase. Our results suggest that As(III) retention during iron reduction is temporally dependent on secondary precipitation of iron phases; during transformation to secondary phases, particularly magnetite, As(III) retention is enhanced even relative to oxidized systems. However, conditions that retard secondary transformation (more stable iron oxides or limited iron reducing bacterial activity), or prolonged anaerobiosis, will lead to both the dissolution of ferric (hydr)oxides and release of As(III) to the aqueous phase.

Oxidative Remobilization of Biogenic Uranium(IV) Precipitates

The oxidative remobilization of uranium from biogenic U(IV) precipitates was investigated in bioreduced sediment suspensions in contact with atmospheric O2 with an emphasis on the influence of Fe(II) and pH on the rate and extent of U release from the solid to the aqueous phase. The sediment was collected from the U.S. Department of Energy Field Research Center (FRC) site at Oak Ridge, Tennessee. Biogenic U(IV) precipitates and bioreduced sediment were generated through anaerobic incubation with a dissimilatory metal reducing bacterium Shewanella putrefaciens strain CN32. The oxidative remobilization of freshly prepared and 1-yr aged biogenic U(IV) was conducted in 0.1 mol/L NaNO3 electrolyte with variable pH and Fe(II) concentrations. Biogenic U(IV)O2(s) was released into the aqueous phase with the highest rate and extent at pH 4 and 9, while the U remobilization was the lowest at circumneutral pH. Increasing Fe(II) significantly decreased U remobilization to the aqueous phase. From 70 to 100% of the U in the sediments used in all the tests was extractable at the experiment termination (41 d) with a bicarbonate solution (0.2 mol/L), indicating that biogenic U(IV) was oxidized regardless of Fe(II) concentration and pH. Sorption experiments and modeling calculations indicated that the inhibitive effect of Fe(II) on U(IV) oxidative remobilization was consistent with the Fe(III) oxide precipitation and U(VI) sorption to this secondary phase.

Microbial reduction of iron(III) oxyhydroxides: effects of mineral solubility and availability

The rate of Fe(III) reductive dissolution by ascorbate has been shown by [Larsen, O. and Postma, D., 2001. Kinetics of reductive bulk dissolution of lepidocrocite, ferrihydrite and goethite. Geochim. Cosmochim. Acta, 65: 1367–1379.] to decrease in the order ferrihydrite>lepidocrocite>goethite>hematite. The abiotic rate of reductive dissolution thus correlates with the solubility of the iron oxyhydroxides. We investigated whether this also holds for the microbial reduction kinetics of Fe(III) iron oxyhydroxides. The solubilities of nanoparticulate hematite, lepidocrocite, ferrihydrite and amorphous Fe(III) oxyhydroxide were obtained from pe–pH titrations of oxyhydroxide/Fe 2+( aq) suspensions, in pH range 4 to 7. The solubility of low surface area (LSA) hematite was estimated from redox potential measurements at pH 2. The same solid phases and soluble Fe(III)-citrate were then reduced by the iron reducing bacterium Shewanella putrefaciens using lactate as the electron donor. In all cases, the microbial Fe(III) reduction rates exhibited saturation behavior with respect to the initial Fe(III) concentration. The maximum specific rate of reduction ( v max, in μmol cell −1 h −1) correlated positively with the solubility of the Fe(III) oxyhydroxides, with the highest values of v max for ferrihydrite and amorphous Fe(III) oxide, the lowest for LSA hematite. Hence, the solubility appears to be a rate-controlling parameter in both the abiotic and enzymatic reduction of Fe(III) oxyhydroxides.

Transformation of 2-line ferrihydrite to 6-line ferrihydrite under oxic and anoxic conditions

Mineralogical transformations of 2-line ferrihydrite were studied under oxic and Fe3+-reducing conditions to establish the role, if any, of 6-line ferrihydrite (“well” organized ferrihydrite) in the reaction pathway and as a final product. In oxic experiments, concentrated suspensions (0.42 mol/L Fe3+ in 0.1 mol/L NaClO4) of freshly synthesized 2-line ferrihydrite, with and without 3% Ni2+, were aged at an initial pH = 7.2 (unbuffered and unadjusted) and 25 °C for more than three years. X-ray diffraction, transmission electron microscopy, and Mossbauer spectroscopy measurements were performed on the solids after different aging periods. The primary mineralogical products observed were 6-line ferrihydrite and goethite, with minor hematite. Aggregation and crystallization of the 2-line ferrihydrite liberated protons and depressed suspension pH, but coprecipitated Ni2+ retarded this process. The joint, interrelated effects of Ni and pH influenced both the extent of conversion of 2-line ferrihydrite and the identity of the major transformation products. Six-line ferrihydrite dominated in the Ni ferrihydrite suspension, whereas goethite dominated in the absence of Ni. Aggregation-induced crystallization of 2-line ferrihydrite particles seemed responsible for 6-line ferrihydrite formation. Mineralogical changes to Ni ferrihydrite under anaerobic conditions were investigated at circumneutral pH using the Fe3+-reducing bacterium Shewanella putrefaciens . Residual 6-line ferrihydrite dominated bioreduced samples that also contained goethite and magnetite. The conversion of 2-line ferrihydrite to 6-line ferrihydrite was considerably more rapid under anaerobic conditions. The sorption of biogenic Fe2+ apparently induced intra-aggregate transformation of 2-line ferrihydrite to 6-line ferrihydrite. Collectively, abiotic and biotic studies indicated that 6-line ferrihydrite can be a transformation product of 2-line ferrihydrite, especially when 2-line ferrihydrite is undergoing transformation to more stable hematite or magnetite.

The roles of natural organic matter in chemical and microbial reduction of ferric iron

Although natural organic matter (NOM) is known to be redox reactive, the roles and effectiveness of specific functional groups of NOM in metal reduction are still a subject of intense investigation. This study entails the investigation of the Fe(III) reduction kinetics and capacity by three fractionated NOM subcomponents in the presence or absence of the dissimilatory metal reducing bacterium Shewanella putrefaciens CN32. Results indicate that NOM was able to reduce Fe(III) abiotically; the reduction was pH-dependent and varied greatly with different fractions of NOM. The polyphenolic-rich NOM-PP fraction exhibited the highest reactivity and oxidation capacity at a low pH (<4) as compared with the carbohydrate-rich NOM-CH fraction and a soil humic acid (soil HA) in reducing Fe(III). However, at a pH>4, soil HA showed a relatively high oxidation capacity, probably resulting from its conformational and solubility changes with an increased solution pH. In the presence of S. putrefaciens CN32, all NOM fractions were found to enhance the microbial reduction of Fe(III) under anaerobic, circumneutral pH conditions. Soil HA was found to be particularly effective in mediating the bioreduction of Fe(III) as compared with the NOM-PP or NOM-CH fractions. NOM-CH was the least effective because it was depleted in both aromatic and polyphenolic organic contents. However, because both soil HA and NOM-PP contain relatively high amounts of aromatic and phenolic compounds, results may indicate that low-molecular-weight polyphenolic organics in NOM-PP were less effective in mediating the bioreduction of Fe(III) at circumneutral pH than the high-molecular-weight polycondensed, conjugated aromatics present in soil HA. These research findings may shed additional light in understanding of the roles and underlying mechanisms of NOM reactions with contaminant metals, radionuclides, and other toxic chemicals in the natural environment.

Crystal Structures of an Oxygen-binding Cytochrome cfrom Rhodobacter sphaeroides

The photosynthetic bacterium Rhodobacter sphaeroides produces a heme protein (SHP), which is an unusual c-type cytochrome capable of transiently binding oxygen during autooxidation. Similar proteins have not only been observed in other photosynthetic bacteria but also in the obligate methylotroph Methylophilus methylotrophus and the metal reducing bacterium Shewanella putrefaciens. A three-dimensional structure of SHP was derived using the multiple isomorphous replacement phasing method. Besides a model for the oxidized state (to 1.82 A resolution), models for the reduced state (2.1 A resolution), the oxidized molecule liganded with cyanide (1. 90 A resolution), and the reduced molecule liganded with nitric oxide (2.20 A resolution) could be derived. The SHP structure represents a new variation of the class I cytochrome c fold. The oxidized state reveals a novel sixth heme ligand, Asn(88), which moves away from the iron upon reduction or when small molecules bind. The distal side of the heme has a striking resemblance to other heme proteins that bind gaseous compounds. In SHP the liberated amide group of Asn(88) stabilizes solvent-shielded ligands through a hydrogen bond.

Microbial Reduction of Iodate

The different oxidation species of iodine have markedly different sorption properties. Hence, changes in iodine redox states can greatly affect the mobility of iodine in the environment. Although a major microbial role has been suggested in the past to account for these redox changes, little has been done to elucidate the responsible microorganisms or the mechanisms involved. In the work presented here, direct microbial reduction of iodate was demonstrated with anaerobic cell suspensions of the sulfate reducing bacterium Desulfovibrio desulfuricans which reduced 96% of an initial 100 µM iodate to iodide at pH 7 in 30 mM NaHCO3 buffer, whereas anaerobic cell suspensions of the dissimilatory Fe(III)-reducing bacterium Shewanella putrefaciens were unable to reduce iodate in 30 mM NaHCO3 buffer (pH 7). Both D. desulfuricans and S. putrefaciens were able to reduce iodate at pH 7 in 10 mM HEPES buffer. Both soluble ferrous iron and sulfide, as well as iron monosulfide (FeS) were shown to abiologically reduce iodate to iodide. These results indicate that ferric iron and/or sulfate reducing bacteria are capable of mediating both direct, enzymatic, as well as abiotic reduction of iodate in natural anaerobic environments. These microbially mediated reactions may be important factors in the fate and transport of129 I in natural systems.

Design and application of rRNA-targeted oligonucleotide probes for the dissimilatory iron- and manganese-reducing bacterium Shewanella putrefaciens.

A 16S rRNA-targeted oligonucleotide probe specific for the iron (Fe3+)- and manganese (Mn4+)-reducing bacterium Shewanella putrefaciens was constructed and tested in both laboratory- and field-based hybridization experiments. The radioactively labeled probe was used to detect S. putrefaciens in field samples collected from the water column and sediments of Oneida Lake in New York and its major southern tributary, Chittenango Creek. S. putrefaciens was quantified by (i) hybridization of the probe to bulk RNA extracted from field samples and normalization of the S. putrefaciens-specific rRNA to total eubacterial rRNA, (ii) a colony-based probe hybridization assay, and (iii) a colony-based biochemical assay which detected the formation of iron sulfide precipitates on triple-sugar iron agar. The results of field applications indicated that the three detection methods were comparable in sensitivity for detecting S. putrefaciens in water column and sediment samples. S. putrefaciens rRNA was detected in the surficial layers of the lake and creek sediments, but the levels of S. putrefaciens rRNA were below the detection limits in the lake and creek water samples. The highest concentrations of S. putrefaciens rRNA, corresponding to approximately 2% of the total eubacterial rRNA, were detected in the surficial sediments of Chittenango Creek and at a midlake site where the Oneida Lake floor is covered by a high concentration of ferromanganese nodules. This finding supports the hypothesis that metal-reducing bacteria such as S. putrefaciens are important components in the overall biogeochemical cycling of iron, manganese and other elements in seasonally anoxic freshwater basins.