Bradley M. Tebo

Occurrence and Mechanisms of Microbial Oxidation of Manganese

Publisher Summary This chapter discusses occurrence and mechanisms of microbial oxidation of Manganese. It examines the biogeochemistry and microbiology of manganese. The chapter focuses on two aspects of the field in which recent progress has been made: field studies of Mn(II) oxidation, including newly developed methods for measuring rates of Mn(II) oxidation and a (2) synopsis of some of the field data that unequivocally establishes the importance of microbes in Mn(II) oxidation in natural systems. The chapter also presents a brief synopsis of some of the field data that unequivocally establishes the importance of microbes in Mn(II) oxidation in natural systems. Finally, the chapter discusses the recent physiological, structural, and biochemical studies of microbial manganese oxidation. In this context, it presents an overview of the chemistry and biology of manganese, which must be understood in order to properly appreciates the field and laboratory studies.

A Large Gene Cluster Encoding Several Magnetosome Proteins Is Conserved in Different Species of Magnetotactic Bacteria

ABSTRACT In magnetotactic bacteria, a number of specific proteins are associated with the magnetosome membrane (MM) and may have a crucial role in magnetite biomineralization. We have cloned and sequenced the genes of several of these polypeptides in the magnetotactic bacterium Magnetospirillum gryphiswaldense that could be assigned to two different genomic regions. Except for mamA, none of these genes have been previously reported to be related to magnetosome formation. Homologous genes were found in the genome sequences ofM. magnetotacticum and magnetic coccus strain MC-1. The MM proteins identified display homology to tetratricopeptide repeat proteins (MamA), cation diffusion facilitators (MamB), and HtrA-like serine proteases (MamE) or bear no similarity to known proteins (MamC and MamD). A major gene cluster containing several magnetosome genes (including mamA and mamB) was found to be conserved in all three of the strains investigated. ThemamAB cluster also contains additional genes that have no known homologs in any nonmagnetic organism, suggesting a specific role in magnetosome formation.

Environmental oxidation rate of manganese(II): bacterial catalysis

Abstract A simple mass balance for dissolved manganese(II) in waters containing low levels of oxygen in Saanich Inlet indicates that the residence time for Mn(II) removal to the solid phase is on the order of a few days. The average oxidation state of Mn in particulate material sampled from the region of Mn removal was 2.3–2.7, and electron micrographs revealed structures characteristic of bacterially formed Mn precipitates. Radiotracer experiments utilizing 54 Mn(II) indicated that removal of Mn from solution in the region of active uptake was substantially blocked by a poison mixture, demonstrating that Mn(II) binding to particulates is catalyzed by bacteria in this environment.

Soluble Mn(III) in Suboxic Zones

Soluble manganese(III) [Mn(III)] has been thought to disproportionate to soluble Mn(II) and particulate MnIVO2 in natural waters, although it persists as complexes in laboratory solutions. We report that, in the Black Sea, soluble Mn(III) concentrations were as high as 5 micromolar and constituted up to 100% of the total dissolved Mn pool. Depth profiles indicated that soluble Mn(III) was produced at the top of the suboxic zone by Mn(II) oxidation and at the bottom of the suboxic zone by MnIVO2 reduction, then stabilized in each case by unknown natural ligands. We also found micromolar concentrations of dissolved Mn(III) in the Chesapeake Bay. Dissolved Mn(III) can maintain the existence of suboxic zones because it can act as either an electron acceptor or donor. Our data indicate that Mn(III) should be ubiquitous at all water column and sediment oxic/anoxic interfaces in the environment.

Structural characterization of biogenic Mn oxides produced in seawater by the marine bacillus sp. strain SG-1

Abstract Natural Mn-oxide nanoparticles and grain coatings are ubiquitous in the environment and profoundly impact the water quality and quality of sediments through their ability to degrade and sequester contaminants. These oxides, which are believed to form dominantly via oxidation of Mn2+ by marine and freshwater bacteria, have extremely high sorptive capacities for heavy metals. We have used XANES, EXAFS, and synchrotron (SR)-XRD techniques to study biogenic Mn oxides produced by spores of the marine Bacillus sp. strain SG-1 in seawater as a function of reaction time under in-situ conditions. An EXAFS model was developed to fully account for the structure and features in the data, providing realistic structural information. The first observed biogenic solid-phase Mn-oxide product is a layered phyllomanganate with hexagonal sheet symmetry and an Mn-oxidation state similar to that in δ-MnO2, between 3.7 and 4.0. XRD and SEM-EDS data show the biooxides to have a phyllomanganate 10 Å basal plane spacing and an interlayer containing Ca. With time, a phyllomanganate oxide with pseudo-orthogonal sheet symmetry appears. Fits to these EXAFS spectra suggest the octahedral layers have relatively few Mn octahedral site vacancies in the lattice and the layers bend to accommodate Jahn-Teller distortions creating the change in symmetry. A reaction mechanism is proposed to account for the observed products. The phyllomanganate oxides observed in this study may be the same as the most abundant Mn-oxide phases suspended in the oxic and sub-oxic zones of the oceanic water column that are of global importance in trace metal and nutrient cycling

Manganese(II) oxidation in the suboxic zone of the Black Sea

Measurements of manganese(II) removal from solution and oxidation were made in the suboxic zone of the Black Sea during Leg 3 of the U.S.-Turkey Black Sea Expedition in June 1988. Two types of rate measurements were conducted. First, Mn(II) removal rate measurements were made under air saturation conditions and in the presence and absence of a biological poison to determine the potential (maximal) rates of Mn(II) binding and oxidation as a result of microbial activity. Second, rates of Mn(II) removal from solution were measured under pH, O2 and temperature conditions simulating those found in situ. By subtracting results from experiments conducted in the absence of oxygen from the oxygen-containing experiments, estimates of Mn(II) oxidation rates could be derived. Both the potential Mn(ll) removal rates and the Mn(II) oxidation rates were 1–2 orders of magnitude higher at the nearshore station (BS3-3) than at a station in the central Black Sea (BS3-6). Residence times of dissolved Mn(II) with respect to oxidation were about 0.6 days at BS3-3 and greater than 9 days at BS3-6. The absolute rates of Mn(II) oxidation in the Black Sea were 1–4 orders of magnitude faster than observed in any other marine environment. The average calculated rate constant for Mn(II) oxidation (kMn) was 1.4 × 1024 M−4 day−1, 5–6 orders of magnitude faster than that calculated for autocatalytic Mn(II) oxidation on the surface of colloidal MnO2. This extremely high rate constant coupled with the inhibition of Mn(II) removal by azide, glutaraldehyde and formaldehyde indicate that Mn(II) oxidation is biologically catalysed. Measurements conducted at BS3-3 1 week apart also demonstrated significant temporal variability in Mn(II) removal rates. Because the removal and oxidation of Mn(II) is so pronounced along the coastal margins it is possible that horizontal advective processes involving Mn(II) oxidation may have a profound influence on the biogeochemistry of other redox sensitive elements within the suboxic zone of the Black Sea and may help explain the depth distribution of particulate Mn observed in the central basin.

Biotic and abiotic products of Mn(II) oxidation by spores of the marine Bacillus sp. strain SG-1

Abstract Bacterial Mn(II) oxidization by spores of Bacillus, sp. strain SG-1 has been systematically probed over the time scale 0.22 to 77 days under in-situ conditions and at differing Mn(II) concentrations. Three complementary techniques, K-edge X-ray absorption near-edge spectroscopy (XANES), X-ray emission spectroscopy (XES), and in-situ synchrotron radiation-based X-ray diffraction (SR-XRD), have been utilized to examine time-dependent changes in Mn oxidation state, local-, and long-range structure in amorphous, crystalline, cell-bound, and solute Mn species. The primary solid biogenic product of Mn(II) oxidation is an X-ray amorphous oxide similar to δ-MnO2, which has a Mn oxidation state between 3.7 and 4.0. Reaction of Mn(II) with the primary biogenic oxide results in the production of abiotic secondary products, feitknechtite or a 10 Å Na phyllomanganate. The identity of the secondary product depends upon the Mn(II) concentration as described by thermodynamic relations. A decrease in the dissolved Mn(II) concentration is followed by mineralogic transformation of the secondary products. Thus, Mn(II) appears to act as a reductant toward the biogenic oxide and to control the stability of secondary reaction products. Mineralogic changes similar to these are likely to be commonplace in natural settings where bacterial Mn(II) oxidation is occurring and may liberate sorbed metal ions or alter the rates of important Mn oxide surface-mediated processes such as the degradation of organic molecules. It is plausible that microbes may exploit such mineral transformation reactions to indirectly control specific chemical conditions in the vicinity of the cell.

Enzymatic manganese(II) oxidation by metabolically dormant spores of diverse Bacillus species

ABSTRACT Bacterial spores are renowned for their longevity, ubiquity, and resistance to environmental insults, but virtually nothing is known regarding whether these metabolically dormant structures impact their surrounding chemical environments. In the present study, a number of spore-forming bacteria that produce dormant spores which enzymatically oxidize soluble Mn(II) to insoluble Mn(IV) oxides were isolated from coastal marine sediments. The highly charged and reactive surfaces of biogenic metal oxides dramatically influence the oxidation and sorption of both trace metals and organics in the environment. Prior to this study, the only known Mn(II)-oxidizing sporeformer was the marine Bacillus sp. strain SG-1, an extensively studied bacterium in which Mn(II) oxidation is believed to be catalyzed by a multicopper oxidase, MnxG. Phylogenetic analysis based on 16S rRNA and mnxG sequences obtained from 15 different Mn(II)-oxidizing sporeformers (including SG-1) revealed extensive diversity within the genus Bacillus, with organisms falling into several distinct clusters and lineages. In addition, active Mn(II)-oxidizing proteins of various sizes, as observed in sodium dodecyl sulfate-polyacrylamide electrophoresis gels, were recovered from the outer layers of purified dormant spores of the isolates. These are the first active Mn(II)-oxidizing enzymes identified in spores or gram-positive bacteria. Although extremely resistant to denaturation, the activities of these enzymes were inhibited by azide and o-phenanthroline, consistent with the involvement of multicopper oxidases. Overall, these studies suggest that the commonly held view that bacterial spores are merely inactive structures in the environment should be revised.

Dissimilatory metal reduction by the facultative anaerobe Pantoea agglomerans SP1.

ABSTRACT Anaerobic enrichments with acetate as the electron donor and Fe(III) as the terminal electron acceptor were obtained from sediments of Salt Pond, a coastal marine basin near Woods Hole, Mass. A pure culture of a facultatively anaerobic Fe(III) reducer was isolated, and 16S rRNA analysis demonstrated that this organism was most closely related to Pantoea (formerly Enterobacter)agglomerans, a member of the familyEnterobacteriaceae within the gamma subdivision of theProteobacteria. This organism, designated strain SP1, can grow by coupling the oxidation of acetate or H2 to the reduction of a variety of electron acceptors, including Fe(III), Mn(IV), Cr(VI), and the humic substance analog 2,6-anthraquinone disulfonate, but not sulfate. To our knowledge, this is the first mesophilic facultative anaerobe reported to couple acetate oxidation to dissimilatory metal reduction.

Bacteriogenic Manganese Oxides

Microorganisms control the redox cycling of manganese in the natural environment. Although the homogeneous oxidation of Mn(II) to form manganese oxide minerals is slow, solid MnO(2) is the stable form of manganese in the oxygenated portion of the biosphere. Diverse bacteria and fungi have evolved the ability to catalyze this process, producing the manganese oxides found in soils and sediments. Other bacteria have evolved to utilize MnO(2) as a terminal electron acceptor in respiration. This Account summarizes the properties of Mn oxides produced by bacteria (bacteriogenic MnO(2)) and our current thinking about the biochemical mechanisms of bacterial Mn(II) oxidation. According to X-ray absorption spectroscopy and X-ray scattering studies, the MnO(2) produced by bacteria consists of stacked hexagonal sheets of MnO(6) octahedra, but these particles are extremely small and have numerous structural defects, particularly cation vacancies. The defects provide coordination sites for binding exogenous metal ions, which can be adsorbed to a high loading. As a result, bacterial production of MnO(2) influences the bioavailability of these metals in the natural environment. Because of its high surface area and oxidizing power, bacteriogenic MnO(2) efficiently degrades biologically recalcitrant organic molecules to lower-molecular-mass compounds, spurring interest in using these properties in the bioremediation of xenobiotic organic compounds. Finally, bacteriogenic MnO(2) is reduced to soluble Mn(II) rapidly in the presence of exogenous ligands or sunlight. It can therefore help to regulate the bioavailability of Mn(II), which is known to protect organisms from superoxide radicals and is required to assemble the water-splitting complex in photosynthetic organisms. Bioinorganic chemists and microbiologists have long been interested in the biochemical mechanism of Mn(IV) oxide production. The reaction requires a two-electron oxidation of Mn(II), but genetic and biochemical evidence for several bacteria implicate multicopper oxidases (MCOs), which are only known to engage one-electron transfers from substrate to O(2). In experiments with the exosporium of a Mn(II)-oxidizing Bacillus species, we could trap the one-electron oxidation product, Mn(III), as a pyrophosphate complex in an oxygen-dependent reaction inhibited by azide, consistent with MCO catalysis. The Mn(III) pyrophosphate complex can further act as a substrate, reacting in the presence of the exosporium to produce Mn(IV) oxide. Although this process appears to be unprecedented in biology, it is reminiscent of the oxidation of Fe(II) to form Fe(2)O(3) in the ferritin iron storage protein. However, it includes a critical additional step of Mn(III) oxidation or disproportionation. We shall continue to investigate this biochemically unique process with purified enzymes.