Scale-up of the production of highly reactive biogenic magnetite nanoparticles using Geobacter sulfurreducens.

Although there are numerous examples of large-scale commercial microbial synthesis routes for organic bioproducts, few studies have addressed the obvious potential for microbial systems to produce inorganic functional biomaterials at scale. Here we address this by focusing on the production of nanoscale biomagnetite particles by the Fe(III)-reducing bacterium Geobacter sulfurreducens, which was scaled up successfully from laboratory- to pilot plant-scale production, while maintaining the surface reactivity and magnetic properties which make this material well suited to commercial exploitation. At the largest scale tested, the bacterium was grown in a 50 l bioreactor, harvested and then inoculated into a buffer solution containing Fe(III)-oxyhydroxide and an electron donor and mediator, which promoted the formation of magnetite in under 24 h. This procedure was capable of producing up to 120 g of biomagnetite. The particle size distribution was maintained between 10 and 15 nm during scale-up of this second step from 10 ml to 10 l, with conserved magnetic properties and surface reactivity; the latter demonstrated by the reduction of Cr(VI). The process presented provides an environmentally benign route to magnetite production and serves as an alternative to harsher synthetic techniques, with the clear potential to be used to produce kilogram to tonne quantities.

Cr(VI) and azo dye removal using a hollow-fibre membrane system functionalized with a biogenic Pd-magnetite catalyst.

This study investigates the application of a hybrid system combining hollow-fibre membrane technology with the reductive abilities of magnetic nanoparticles for the remediation of toxic Cr(VI) and the azo dye, Remazol Black B. Nano-scale biogenic magnetite (Fe3O4), formed by microbial reduction of the mineral ferrihydrite, has a high reductive capacity due to the presence of Fe(II) in the mineral structure. The magnetic nanoparticles (approximately 20 nm) can be arrayed with Pd0 nanoparticles (approximately 5 nm) making a catalytically active nanomaterial. Membrane units, with and without nanoparticles, were challenged with either Cr(VI) or azo dye and some were supplemented with sodium formate, as an electron donor for contaminant reduction promoted by the Pd. The combination of Pd-magnetite with formate resulted in the most effective remediation strategy for both contaminants and the lifetime of the membrane unit was also increased, with 55% (19 days) and 70% (23 days) removal of the azo dye and Cr(VI), respectively. Low flow rates of 0.1 ml/min resulted in improved efficiencies due to increased contact time with the membrane/nanoparticle unit, with 70-75% removal of each contaminant. Chemical analyses of the nanoparticles post-exposure to Cr(VI) in the membrane modules indicated Pd to be more oxidized when Cr removal was maximized, and that the Cr was partially reduced to Cr(III) at the surface of the magnetite. These results have demonstrated that hollow-fibre membrane units can be enhanced for the removal of soluble, redox sensitive contaminants by incorporation of a layer of palladized biogenic nanoparticulate magnetite.

Controlled cobalt doping in biogenic magnetite nanoparticles.

Cobalt-doped magnetite (CoxFe3 -xO4) nanoparticles have been produced through the microbial reduction of cobalt-iron oxyhydroxide by the bacterium Geobacter sulfurreducens. The materials produced, as measured by superconducting quantum interference device magnetometry, X-ray magnetic circular dichroism, Mössbauer spectroscopy, etc., show dramatic increases in coercivity with increasing cobalt content without a major decrease in overall saturation magnetization. Structural and magnetization analyses reveal a reduction in particle size to less than 4 nm at the highest Co content, combined with an increase in the effective anisotropy of the magnetic nanoparticles. The potential use of these biogenic nanoparticles in aqueous suspensions for magnetic hyperthermia applications is demonstrated. Further analysis of the distribution of cations within the ferrite spinel indicates that the cobalt is predominantly incorporated in octahedral coordination, achieved by the substitution of Fe(2+) site with Co(2+), with up to 17 per cent Co substituted into tetrahedral sites.

Ex situ formation of metal selenide quantum dots using bacterially derived selenide precursors.

Luminescent quantum dots were synthesized using bacterially derived selenide (Se(II-)) as the precursor. Biogenic Se(II-) was produced by the reduction of Se(IV) by Veillonella atypica and compared directly against borohydride-reduced Se(IV) for the production of glutathione-stabilized CdSe and ß-mercaptoethanol-stabilized ZnSe nanoparticles by aqueous synthesis. Biological Se(II-) formed smaller, narrower size distributed QDs under the same conditions. The growth kinetics of biologically sourced CdSe phases were slower. The proteins isolated from filter sterilized biogenic Se(II-) included a methylmalonyl-CoA decarboxylase previously characterized in the closely related Veillonella parvula. XAS analysis of the glutathione-capped CdSe at the S K-edge suggested that sulfur from the glutathione was structurally incorporated within the CdSe. A novel synchrotron based XAS technique was also developed to follow the nucleation of biological and inorganic selenide phases, and showed that biogenic Se(II-) is more stable and more resistant to beam-induced oxidative damage than its inorganic counterpart. The bacterial production of quantum dot precursors offers an alternative, 'green' synthesis technique that negates the requirement of expensive, toxic chemicals and suggests a possible link to the exploitation of selenium contaminated waste streams.

Characterisation of the dissimilatory reduction of Fe(III)-oxyhydroxide at the microbe-mineral interface: the application of STXM-XMCD.

A combination of scanning transmission X-ray microscopy and X-ray magnetic circular dichroism was used to spatially resolve the distribution of different carbon and iron species associated with Shewanella oneidensis MR-1 cells. S. oneidensis MR-1 couples the reduction of Fe(III)-oxyhydroxides to the oxidation of organic matter in order to conserve energy for growth. Several potential mechanisms may be used by S. oneidensis MR-1 to facilitate Fe(III)-reduction. These include direct contact between the cell and mineral surface, secretion of either exogenous electron shuttles or Fe-chelating agents and the production of conductive 'nanowires'. In this study, the protein/lipid signature of the bacterial cells was associated with areas of magnetite (Fe3O4), the product of dissimilatory Fe(III) reduction, which was oversaturated with Fe(II) (compared to stoichiometric magnetite). However, areas of the sample rich in polysaccharides, most likely associated with extracellular polymeric matrix and not in direct contact with the cell surface, were undersaturated with Fe(II), forming maghemite-like (γ-Fe2O3) phases compared to stoichiometric magnetite. The reduced form of magnetite will be much more effective in environmental remediation such as the immobilisation of toxic metals. These findings suggest a dominant role for surface contact-mediated electron transfer in this study and also the inhomogeneity of magnetite species on the submicron scale present in microbial reactions. This study also illustrates the applicability of this new synchrotron-based technique for high-resolution characterisation of the microbe-mineral interface, which is pivotal in controlling the chemistry of the Earth's critical zone.

Control of nanoparticle size, reactivity and magnetic properties during the bioproduction of magnetite by Geobacter sulfurreducens.

The bioproduction of nanoscale magnetite by Fe(III)-reducing bacteria offers a potentially tunable, environmentally benign route to magnetic nanoparticle synthesis. Here, we demonstrate that it is possible to control the size of magnetite nanoparticles produced by Geobacter sulfurreducens by adjusting the total biomass introduced at the start of the process. The particles have a narrow size distribution and can be controlled within the range of 10-50 nm. X-ray diffraction analysis indicates that controlled production of a number of different biominerals is possible via this method including goethite, magnetite and siderite, but their formation is strongly dependent upon the rate of Fe(III) reduction and total concentration and rate of Fe(II) produced by the bacteria during the reduction process. Relative cation distributions within the structure of the nanoparticles have been investigated by x-ray magnetic circular dichroism and indicate the presence of a highly reduced surface layer which is not observed when magnetite is produced through abiotic methods. The enhanced Fe(II)-rich surface, combined with small particle size, has important environmental applications such as in the reductive bioremediation of organics, radionuclides and metals. In the case of Cr(VI), as a model high-valence toxic metal, optimized biogenic magnetite is able to reduce and sequester the toxic hexavalent chromium very efficiently to the less harmful trivalent form.

Biomineralization: linking the fossil record to the production of high value functional materials.

The microbial cell offers a highly efficient template for the formation of nanoparticles with interesting properties including high catalytic, magnetic and light-emitting activities. Thus biomineralization products are not only important in global biogeochemical cycles, but they also have considerable commercial potential, offering new methods for material synthesis that eliminate toxic organic solvents and minimize expensive high-temperature and pressure processing steps. In this review we describe a range of bacterial processes that can be harnessed to make precious metal catalysts from waste streams, ferrite spinels for biomedicine and catalysis, metal phosphates for environmental remediation and biomedical applications, and biogenic selenides for a range of optical devices. Recent molecular-scale studies have shown that the structure and properties of bionanominerals can be fine-tuned by subtle manipulations to the starting materials and to the genetic makeup of the cell. This review is dedicated to the late Terry Beveridge who contributed much to the field of biomineralization, and provided early models to rationalize the mechanisms of biomineral synthesis, including those of geological and commercial potential.

XAS and XMCD evidence for species-dependent partitioning of arsenic during microbial reduction of ferrihydrite to magnetite.

Poorly crystalline Fe(III) oxyhydroxides, ubiquitously distributed as mineral coatings and discrete particles in aquifer sediments, are well-known hosts of sedimentary As. Microbial reduction of these phases is widely thought to be responsible for the genesis of As-rich reducing groundwaters found in many parts of the world, most notably in Bangladesh and West Bengal, India. As such, it is important to understand the behavior of As associated with ferric oxyhydroxides during the early stages of Fe(lll) reduction. We have used X-ray absorption spectroscopy (XAS) and X-ray magnetic circular dichroism (XMCD) to elucidate the changes in the bonding mechanism of As(III) and As(V) as their host Fe(III) oxyhydroxide undergoes bacterially induced reductive transformation to magnetite. Two-line ferrihydrite, with adsorbed As(III) or As(V), was incubated under anaerobic conditions in the presence of acetate as an electron donor, and Geobacter sulfurreducens, a subsurface bacterium capable of respiring on Fe(lll), but not As(V). In both experiments, no increase in dissolved As was observed during reduction to magnetite (complete upon 5 days incubation), consistent with our earlier observation of As sequestration by the formation of biogenic Fe(III)-bearing minerals. XAS data suggested that the As bonding environment of the As(III)-magnetite product is indistinguishable from that obtained from simple adsorption of As(lll) on the surface of biogenic magnetite. In contrast, reduction of As(V)-sorbed ferrihydrite to magnetite caused incorporation of As5+ within the magnetite structure. XMCD analysis provided further evidence of structural partitioning of As5+ as the small size of the As5+ cation caused a distortion of the spinel structure compared to standard biogenic magnetite. These results may have implications regarding the species-dependent mobility of As undergoing anoxic biogeochemical transformations, e.g., during early sedimentary diagenesis.