Bioreduction Of Lepidocrocite Research Articles

Electron Donor Utilization and Secondary Mineral Formation during the Bioreduction of Lepidocrocite by Shewanella putrefaciens CN32

The bioreduction of Fe(III) oxides by dissimilatory iron reducing bacteria (DIRB) may result in the production of a suite of Fe(II)-bearing secondary minerals, including magnetite, siderite, vivianite, green rusts, and chukanovite; the formation of specific phases controlled by the interaction of various physiological and geochemical factors. In an effort to better understand the effects of individual electron donors on the formation of specific Fe(II)-bearing secondary minerals, we examined the effects of a series of potential electron donors on the bioreduction of lepidocrocite (γ-FeOOH) by Shewanella putrefaciens CN32. Biomineralization products were identified by X-ray diffraction, Mössbauer spectroscopy, and scanning electron microscopy. Acetate, citrate, ethanol, glucose, glutamate, glycerol, malate, and succinate were not effectively utilized for the bioreduction of lepidocrocite by S. putrefaciens CN32; however, substantial Fe(II) production was observed when formate, lactate, H2, pyruvate, serine, or N acetylglucosamine (NAG) was provided as an electron donor. Carbonate or sulfate green rust was the dominant Fe(II)-bearing secondary mineral when formate, H2, lactate, or NAG was provided, however, siderite formed with pyruvate or serine. Geochemical modeling indicated that pH and carbonate concentration are the key factors determining the prevalence of carbonate green rust verses siderite.

Magnetic comparison of abiogenic and biogenic alteration products of lepidocrocite

Lepidocrocite is a potentially important Fe-bearing precursor phase for the production of nanoscale Fe-oxide particles in the environment. We present a detailed magnetic characterization of various alteration products of lepidocrocite resulting from thermal dehydroxylation reactions and bacterially induced bioreduction and remineralization, accompanied by characterization with x-ray diffraction (XRD) and transmission electron microscopy. Dehydroxylation during annealing at moderate temperatures produces a topotactic transformation from lepidocrocite to maghemite when heated in an oxidizing atmosphere, or to magnetite when heated in a reducing atmosphere. The abiotic Fe-oxide products form an oriented framework of strongly interacting superparamagnetic crystallites and are characterized by a distinctive porous nanostructure observed by electron microscopy. Lepidocrocite bioreduction by the iron-reducing bacterium Shewanella putrefaciens ATCC 8071 produces nanoscale particles of a strongly magnetic phase. This Fe(II)-bearing mineral produced by bioreduction is highly crystalline and euhedral in shape, with a broad grain size distribution and is indicated by magnetic and XRD measurements to be a cation-excess magnetite. We highlight the distinguishing microscopic characteristics of magnetite from both abiotic and bacterially induced mineralization that should allow them to be identified in natural settings. Moreover, both mechanisms of alteration represent potential pathways for the direct formation of strongly magnetic fine-grained Fe-oxide particles in sedimentary environments.

Nitrite Reduction by Biogenic Hydroxycarbonate Green Rusts: Evidence for Hydroxy-nitrite Green Rust Formation as an Intermediate Reaction Product

The present study investigates for the first time the reduction of nitrite by biogenic hydroxycarbonate green rusts, bio-GR(CO3), produced from the bioreduction of ferric oxyhydroxycarbonate (Fohc), a poorly crystalline solid phase, and of lepidocrocite, a well-crystallized Fe(III)-oxyhydroxide mineral. Results show a fast Fe(II) production from Fohc, which leads to the precipitation of bio-GR(CO3) particles that were roughly 2-fold smaller (2.3 ± 0.4 μm) than those obtained from the bioreduction of lepidocrocite (5.0 ± 0.4 μm). The study reveals that both bio-GR(CO3) are capable of reducing nitrite ions into gaseous nitrogen species such as NO, N2O, or N2 without ammonium production at neutral initial pH and that nitrite reduction proceeded to a larger extent with smaller particles than with larger ones. On the basis of the identification of intermediates and end-reaction products using X-ray diffraction and X-ray absorption fine structure (XAFS) spectroscopy at the Fe K-edge, our study shows the formation of hydroxy-nitrite green rust, GR(NO2), a new type of green rust 1, and suggests that the reduction of nitrite by biogenic GR(CO3) involves both external and internal reaction sites and that such a mechanism could explain the higher reactivity of green rust with respect to nitrite, compared to other mineral substrates possessing only external reactive sites.

Contribution of Anionic vs. Neutral Polymers to the Formation of Green Rust 1 from γ-FeOOH Bioreduction

To assess the effect of polymeric substances on the biomineralization and stabilization of green rust (GR), the effect of two organic polymers on the transformation of lepidocrocite (γ-FeOOH) to GR vs. magnetite in presence of Shewanella putrefaciens was investigated. These two polymers, generally used as flocculants, are polyacrylic acid (PAA), which bears negatively charged carboxylic groups at neutral pH and is expected to react with cationic hydrolyzed iron species, and polyacrylamide (PAM), which is a neutral polymer that may develop hydrogen bonds with iron nanocolloids. The bioreduction of lepidocrocite by S. putrefaciens was performed under conditions known to yield either magnetite or GR. Each operational condition of interest was investigated with various polymer concentrations from 0.6 to 60 mg g−1 Fe (10 to 1000 mg l−1). The final product was characterized using X-ray diffraction and electronic microscopy. The results showed that the formation of GR is favored, with respect to magnetite, to a lesser extent with PAM than with PAA. These results indicated that polymers influence the chemical stability of GR and/or guide the route of biomineralization. Polymer properties, in addition to silica and phosphate concentrations, are then critical parameters that control the secondary iron mineral biomineralization from iron-reducing bacteria. Supplemental materials are available for this article. Go to the publisher's online edition of Geomicrobiology Journal to view the supplemental file.

Biotransformation of lepidocrocite in the presence of quinones and flavins

This study investigated the bioreduction of lepidocrocite (γ-FeIIIOOH) and its mineral transformation in the presence of exogenous (quinones) and endogenous (flavins) electron transfer mediators (ETMs) at low concentrations of the ETMs and bacterial cells (Shewanella putrefaciens CN32). It is very important to investigate the bioreduction of lepidocrocite in the presence of different ETMs because biotransformation of Fe(III)-containing minerals can be stimulated by ETMs and affect fate and transport of contaminants in contaminated environments. In the absence of phosphate, green rust formation was observed with fast Fe(II) production rate (0.44–0.56mMd−1) during the bioreduction of lepidocrocite with exogenous ETMs, while goethite formed at slow Fe(II) production rate (0.24–0.29mMd−1) with endogenous ETMs. In the presence of phosphate, formation of green rust and vivianite was observed with fast Fe(II) production rate (0.54–0.74mMd−1) during the bioreduction of lepidocrocite with exogenous ETMs, while vivianite formed at moderate Fe(II) production rate (0.36–0.40mMd−1) with endogenous ETMs. Vivianite formed in all experimental cases with phosphate in a broad range of Fe(II) production rates (0.23–0.74mMd−1). Our results (1) suggest that exogenous and endogenous ETMs can significantly but differently affect the biotransformation of lepidocrocite, especially at low concentrations of the ETMs and bacterial cells, (2) highlight the importance of Fe(II) production rate to determine the formation of specific biogenic minerals, (3) provide additional evidence that phosphate can significantly affect the bioreduction rate and the mineral transformation, and (4) help to understand the basic knowledge about complex interactions among microbial cell, soil mineral, and ETM in natural environments and engineered systems.

In situ monitoring of lepidocrocite bioreduction and magnetite formation by reflection Mossbauer spectroscopy

The miniaturized Mössbauer spectrometer (MIMOS II) was used to monitor in situ the mineralogical transformation of lepidocrocite (γ-FeOOH) in a Shewanella putrefaciens culture under anaerobic conditions using methanoate as the electron source. Magnetite was the only biogenic mineral formed during the course of the incubation. The analysis of the biogenic mineral by transmission electron microscopy (TEM) revealed cubic-shaped crystals with a relatively homogeneous grain size of about 50 nm. After one day of incubation, the departure from stoichiometry, δ, of the biogenerated magnetite was very low (δ ~ 0.025) and rapidly reached values close to zero. Such low values of δ were not obtained for magnetite synthesized inorganically when Fe3+ in the form of γ-FeOOH was reacted with stoichiometric quantities of soluble Fe2+ and OH-. The experimental setup used in this study could be replicated in field experiments when assessing the formation of magnetite in modern geological settings as its formation is suspected to be caused by a strong bacterial activity.

Effects of Si-bearing minerals on the nature of secondary iron mineral products from lepidocrocite bioreduction

Lepidocrocite (γ-FeOOH) bioreduction by Shewanella putrefaciens was examined in systems containing three kinds of quartz silica sand with solid contents ranging from 5 to 70 wt.%. The results notably demonstrated that the formation routes of secondary iron minerals from microbial reduction of γ-FeOOH in silica mineral assemblages are dependent on quartz sand loading and particle size distribution. Biomineralization experiments with a concentration range of aqueous silica showed that a lower Si concentration (less than 2 mM) led to magnetite or magnetite/chukanovite, while higher concentrations resulted in the formation of green rust (GR). Dissolution of very fine quartz particles may have promoted the formation of biogenic GR instead of magnetite. This behavior could be explained by the sorption of dissolved silicate on the lepidocrocite surface, which might limit the bioreduction rate and promote GR formation. As observed with the chemically synthesized GR, the interactions of dissolved or colloidal Si with the lateral faces of the GR crystals could stabilize the GR structure, thus preventing its transformation into other products. Furthermore, complex physical and chemical distribution at the surfaces and within heteroaggregates of mixed mineral phases may affect the diffusion processes of species and therefore the biomineralization mechanism.

Effects of Oxyanions, Natural Organic Matter, and Bacterial Cell Numbers on the Bioreduction of Lepidocrocite (γ-FeOOH) and the Formation of Secondary Mineralization Products

Microbial reduction of Fe(III) oxides results in the production of Fe(II) and may lead to the subsequent formation of Fe(II)-bearing secondary mineralization products including magnetite, siderite, vivianite, chukanovite (ferrous hydroxy carbonate (FHC)), and green rust; however, the factors controlling the formation of specific Fe(II) phases are often not well-defined. This study examined effects of (i) a range of inorganic oxyanions (arsenate, borate, molybdate, phosphate, silicate, and tungstate), (ii) natural organic matter (citrate, oxalate, microbial extracellular polymeric substances [EPS], and humic substances), and (iii) the type and number of dissimilatory iron-reducing bacteria on the bioreduction of lepidocrocite and formation of Fe(II)-bearing secondary mineralization products. The bioreduction kinetics clustered into two distinct Fe(II) production profiles. "Fast" Fe(II) production kinetics [19-24 mM Fe(II) d(-1)] were accompanied by formation of magnetite and FHC in the unamended control and in systems amended with borate, oxalate, gellan EPS, or Pony Lake fulvic acid or having "low" cell numbers. Systems amended with arsenate, citrate, molybdate, phosphate, silicate, tungstate, EPS from Shewanella putrefaciens CN32, or humic substances derived from terrestrial plant material or with "high" cell numbers exhibited comparatively slow Fe(II) production kinetics [1.8-4.0 mM Fe(II) d(-1)] and the formation of green rust. The results are consistent with a conceptual model whereby competitive sorption of more strongly bound anions blocks access of bacterial cells and reduced electron-shuttling compounds to sites on the iron oxide surface, thereby limiting the rate of bioreduction.

Bacterial and iron oxide aggregates mediate secondary iron mineral formation: green rust versus magnetite

In the presence of methanoate as electron donor, Shewanella putrefaciens, a Gram-negative, facultative anaerobe, is able to transform lepidocrocite (gamma-FeOOH) to secondary Fe (II-III) minerals such as carbonated green rust (GR1) and magnetite. When bacterial cells were added to a gamma-FeOOH suspension, aggregates were produced consisting of both bacteria and gamma-FeOOH particles. Recently, we showed that the production of secondary minerals (GR1 vs. magnetite) was dependent on bacterial cell density and not only on iron reduction rates. Thus, gamma-FeOOH and S. putrefaciens aggregation pattern was suggested as the main mechanism driving mineralization. In this study, lepidocrocite bioreduction experiments, in the presence of anthraquinone disulfonate, were conducted by varying the [cell]/[lepidocrocite] ratio in order to determine whether different types of aggregate are formed, which may facilitate precipitation of GR1 as opposed to magnetite. Confocal laser scanning microscopy was used to analyze the relative cell surface area and lepidocrocite concentration within the aggregates and captured images were characterized by statistical methods for spatial data (i.e. variograms). These results suggest that the [cell]/[lepidocrocite] ratio influenced both the aggregate structure and the nature of the secondary iron mineral formed. Subsequently, a [cell]/[lepidocrocite] ratio above 1 x 10(7) cells mmol(-1) leads to densely packed aggregates and to the formation of GR1. Below this ratio, looser aggregates are formed and magnetite was systematically produced. The data presented in this study bring us closer to a more comprehensive understanding of the parameters governing the formation of minerals in dense bacterial suspensions and suggest that screening mineral-bacteria aggregate structure is critical to understanding (bio)mineralization pathways.

Arsenite sequestration at the surface of nano-Fe(OH) 2, ferrous-carbonate hydroxide, and green-rust after bioreduction of arsenic-sorbed lepidocrocite by Shewanella putrefaciens

X-ray Absorption Fine Structure (XAFS) spectroscopy was used in combination with high resolution transmission electron microscopy (HRTEM), electron energy loss spectroscopy (EELS), X-ray energy dispersive spectroscopy (XEDS), X-ray powder diffraction, and Mössbauer spectroscopy to obtain detailed information on arsenic and iron speciation in the products of anaerobic reduction of pure and As(V)- or As(III)-adsorbed lepidocrocite (γ-FeOOH) by Shewanella putrefaciens ATCC 12099. We found that this strain of S. putrefaciens is capable of using Fe(III) in lepidocrocite and As(V) in solution or adsorbed on lepidocrocite surfaces as electron acceptors. Bioreduction of lepidocrocite in the absence of arsenic resulted in the formation of hydroxycarbonate green rust 1 [Fe II 4Fe III 2(OH) 12CO 3: GR1(CO 3)], which completely converted into ferrous-carbonate hydroxide (Fe II 2(OH) 2CO 3: FCH) over nine months. This study thus provides the first evidence of bacterial reduction of stoichiometric GR1(CO 3) into FCH. Bioreduction of As(III)-adsorbed lepidocrocite also led to the formation of GR1(CO 3) prior to formation of FCH, but the presence of As(III) slows down this transformation, leading to the co-occurrence of both phases after 22-month of aging. At the end of this experiment, As(III) was found to be adsorbed on the surfaces of GR1(CO 3) and FCH. After five months, bioreduction of As(V)-bearing lepidocrocite led directly to the formation of FCH in association with nanometer-sized particles of a minor As-rich Fe(OH) 2 phase, with no evidence for green rust formation. In this five-month experiment, As(V) was fully converted to As(III), which was dominantly sorbed at the surface of the Fe(OH) 2 nanoparticles as oligomers binding to the edges of Fe(OH) 6 octahedra at the edges of the octahedral layers of Fe(OH) 2. These multinuclear As(III) surface complexes are characterized by As–As pairs at a distance of 3.32 ± 0.02 Å and by As–Fe pairs at a distance of 3.50 ± 0.02 Å and represent a new type of As(III) surface complex. Chemical analyses show that the majority of As(III) produced in the experiments with As present is associated with iron-bearing hydroxycarbonate or hydroxide solids, reinforcing the idea that, at least under some circumstances, bacterial reduction can promote As(III) sequestration instead of mobilizing it into solution.

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.

Green Rust Formation from the Bioreduction of γ –FeOOH (Lepidocrocite): Comparison of Several Shewanella Species

Green rusts are mixed ferrous/ferric hydroxides that typically form under weakly acidic to alkaline conditions in suboxic environments. The recent identification of green rusts as products of the reduction of Fe(III) oxides and oxyhydroxides by Shewanella putrefaciens, a dissimilatory iron-reducing bacterium (DIRB), suggests that green rusts may play a role in the redox cycling of Fe in many aquatic and terrestrial environments. We examined the potential for green rust formation resulting from the bioreduction of lepidocrocite(γ -FeOOH) by a series of Shewanella species (S. alga BrY, S. amazonensis SB2B, S. baltica OS155, S. denitrificans OS217T, S. loihica PV-4, S. oneidensis MR-1, S. putrefaciens ATCC 8071, S. putrefaciens CN32, S. saccharophilia, and Shewanella sp. ANA-3). All Shewanella species, with the exception of S. denitrificans OS217T, were able to couple the oxidation of formate to the reduction of Fe(III) in lepidocrocite; however there were significant differences among species with respect to the rate and extent of Fe(II) production. Despite these differences, green rust was the only Fe(II)-bearing solid phase formed under our experimental conditions, as indicated by X-ray diffraction, Mössbauer spectroscopy, and scanning electron microscopy. The formation of green rust by Shewanella species isolated from a wide range of habitats and possessing varied metabolic capabilities suggests that under favorable conditions biogenic green rusts may be formed by a diverse array of DIRB.

Monitoring structural transformation of hydroxy-sulphate green rust in the presence of sulphate reducing bacteria

The activities of bacterial consortia enable organisms to maximize their metabolic capabilities. This article assesses the synergetic relationship between iron reducing bacteria (IRB), Shewanella putrefaciens and sulphate reducing bacteria (SRB) Desulfovibrio alaskensis. Thus, the aim of this study was first to form a biogenic hydroxy-sulpahte green rust GR2(\( {\text{SO}}_{{\text{4}}} ^{{2 - }} \)) through the bioreduction of lepidocrocite by S. putrefaciens and secondly to investigate if sulfate anions intercalated in the biogenic GR2(\( {\text{SO}}_{{\text{4}}} ^{{2 - }} \)) could serve as final electron acceptor for a sulfate reducing bacterium, D. alaskensis. The results indicate that the IRB lead to the formation of GR2(\( {\text{SO}}_{{\text{4}}} ^{{2 - }} \)) and this mineral serve as an electron acceptor for SRB. GR2(\( {\text{SO}}_{{\text{4}}} ^{{2 - }} \)) precipitation and its transformation was demonstrated by using X-ray diffraction (DRX), Mossbauer spectroscopy (TMS) and transmission electron spectroscopy (TEM). These observations point out the possible acceleration of steel corrosion in marine environment in presence of IRB/SRB consortia.

Iron(II,III) hydroxycarbonate green rust formation and stabilization from lepidocrocite bioreduction.

Bioreduction of the well-crystallized ferric oxyhydroxide gamma-FeOOH lepidocrocite was investigated in batch cultures using Shewanella putrefaciens bacterium (strain CIP 8040) at initial pH 7.5 in bicarbonate buffer. The cultures were performed with formate as electron donor without phosphate, in the presence or absence of anthraquinone-2,6-disulfonate (AQDS) as electron shuttle. During lepidocrocite reduction, the iron(II,III) hydroxycarbonate green rust GR(CO32-) was characterized by X-ray diffraction, transmission electron microscopy, and transmission Mössbauer spectroscopy. The AQDS accelerated the kinetics of GR formation. GR was the major end product when bacterial reduction was not stopped by lack of electron donor, and between 55 and 86% of the iron from gamma-FeOOH precipitated in GR(CO32-). However, when the bacterial reduction was stopped by freezing/thawing or the electron donor was exhausted, the large quantity of remaining lepidocrocite induced a transformation of GR into magnetite. This confirms that GR is metastable with respect to magnetite in the presence of gamma-FeOOH.