Investigating Abiotic and Biotic Mechanisms of Pyrite Reduction

Abstract

Pyrite (FeS2) has a very low solubility and therefore has historically been considered a sink for iron (Fe) and sulfur (S) and unavailable to biology in the absence of oxygen and oxidative weathering. Anaerobic methanogens were recently shown to reduce FeS2 and assimilate Fe and S reduction products to meet nutrient demands. However, the mechanism of FeS2 mineral reduction and the forms of Fe and S assimilated by methanogens remained unclear. Thermodynamic calculations described herein indicate that H2 at aqueous concentrations as low as 10–10 M favors the reduction of FeS2, with sulfide (HS–) and pyrrhotite (Fe1–xS) as products; abiotic laboratory experiments confirmed the reduction of FeS2 with dissolved H2 concentrations greater than 1.98 × 10–4 M H2. Growth studies of Methanosarcina barkeri provided with FeS2 as the sole source of Fe and S resulted in H2 production but at concentrations too low to drive abiotic FeS2 reduction, based on abiotic laboratory experimental data. A strain of M. barkeri with deletions in all [NiFe]-hydrogenases maintained the ability to reduce FeS2 during growth, providing further evidence that extracellular electron transport (EET) to FeS2 does not involve H2 or [NiFe]-hydrogenases. Physical contact between cells and FeS2 was required for mineral reduction but was not required to obtain Fe and S from dissolution products. The addition of a synthetic electron shuttle, anthraquinone-2,6-disulfonate, allowed for biological reduction of FeS2 when physical contact between cells and FeS2 was prohibited, indicating that exogenous electron shuttles can mediate FeS2 reduction. Transcriptomics experiments revealed upregulation of several cytoplasmic oxidoreductases during growth of M. barkeri on FeS2, which may indicate involvement in provisioning low potential electrons for EET to FeS2. Collectively, the data presented herein indicate that reduction of insoluble FeS2 by M. barkeri occurred via electron transfer from the cell surface to the mineral surface resulting in the generation of soluble HS– and mineral-associated Fe1–xS. Solubilized Fe(II), but not HS–, from mineral-associated Fe1–xS reacts with aqueous HS– yielding aqueous iron sulfur clusters (FeSaq) that likely serve as the Fe and S source for methanogen growth and activity. FeSaq nucleation and subsequent precipitation on the surface of cells may result in accelerated EET to FeS2, resulting in positive feedback between cell activity and FeS2 reduction.

A naturalist perspective of microbiology: Examples from methanogenic archaea

“…Intriguing recent studies of the methanogens Methanococcus voltae (Methanococcales) and Methanosarcina barkeri (Methanosarcinales) showed that they can reductively dissolve FeS 2 and use dissolution products to meet Fe and S demands (Payne et al, 2021;Spietz, Payne, Kulkarni, et al, 2022b). Methanococcus voltae encodes a SUF-minimal pathway lacking a homologue of cysteine desulfurase whereas M. barkeri encodes a traditional SUF pathway with a homologue of cysteine desulfurase (Johnson et al, 2021).…”

“…Recent studies that showed FeS (aq) clusters are made available through FeS 2 reduction (Payne et al, 2021; Spietz, Payne, Kulkarni, et al, 2022b) and are likely neutrally charged at circumneutral pH (Rickard & Luther, 2007). FeS (aq) are likely to be directly assimilated by methanogens, which provides a new avenue to begin to explore questions of how such a transition between abiotic catalysis and biotic catalysis might have taken place.…”

“…Methanococcus voltae encodes a SUF‐minimal pathway lacking a homologue of cysteine desulfurase whereas M. barkeri encodes a traditional SUF pathway with a homologue of cysteine desulfurase (Johnson et al, 2021). Based on available chemical and microbial data from growth studies, Spietz et al describe a model for reductive dissolution of FeS 2 wherein FeS 2 reduction occurs via spontaneous extracellular electron transfer from the cell membrane to the mineral, with direct cell‐mineral contact required for reduction (Spietz, Payne, Kulkarni, et al, 2022b; Spietz, Payne, Szilagyi, & Boyd, 2022a). Reduction of a surface exposed persulfide in FeS 2 releases HS − into solution and generates pyrrhotite (Fe 1‐ x S) as a residual mineral on the surface of FeS 2 .…”

Development of molecular cluster models to probe pyrite surface reactivity

“…However, several recent studies have shown that FeS 2(p) can be reductively dissolved through both abiotic and biotic processes at anoxic, physiological conditions 5 . For the former, dissolved hydrogen gas in micromolar concentrations and at biologically relevant temperatures (approximately 40°C) can abiotically reduce FeS 2(p) , resulting in what has been hypothesized to be a pyrrhotite (Fe 1‐x S (p) ) surface‐associated phase and release of HS − (aq) into solution 6 . For the latter, methanogens as anaerobic archaea generating methane via metabolism of carbon dioxide, alcohols, and organic acids, 7 were also shown to reduce FeS 2(p) 8–13 .…”

Proteomic Analysis of Methanococcus voltae Grown in the Presence of Mineral and Nonmineral Sources of Iron and Sulfur

“…The mechanism and cellular pathways responsible for this process have yet to be elucidated. Recent transcriptomics work on Methanosarcina barkeri cultured in the presence of different Fe/S sources implicates alpha-keto reductases, a flavin mononucleotide-dependent flavodoxin reductase, and hydrolases as putative enzymes involved in FeS 2 reduction ( 21 ). Our previous work with M. voltae showed that cells grown on FeS 2 are smaller and may use an IssA protein to sequester iron as a thioferrate-like species ( 20 ).…”