New Results

H2 is a Major Intermediate in Desulfovibrio vulgaris Corrosion of Iron

Trevor L. Woodard, Toshiyuki Ueki, Derek R. Lovley
doi: https://doi.org/10.1101/2022.11.15.516606

Abstract

Desulfovibrio vulgaris has been the primary pure culture sulfate reducer for developing microbial corrosion concepts. Multiple mechanisms for how it accepts electrons from Fe0 have been proposed. We investigated Fe0 oxidation with a mutant of D. vulgaris in which hydrogenase genes were deleted. The hydrogenase mutant grew as well as the parental strain with lactate as the electron donor, but unlike the parental strain was not able to grow on H2. The parental strain reduced sulfate with Fe0 as the sole electron donor, but the hydrogenase mutant did not. H2 accumulated over time in Fe0 cultures of the hydrogenase mutant and sterile controls, but not in parental strain cultures. Sulfide stimulated H2 production in uninoculated controls apparently by both reacting with Fe0 to generate H2 and facilitating electron transfer from Fe0 to H+. Parental strain supernatants did not accelerate H2 production from Fe0, ruling out a role for extracellular hydrogenases. Previously proposed electron transfer between Fe0 and D. vulgaris via soluble electron shuttles was not evident. The hydrogenase mutant did not reduce sulfate in the presence of Fe0 and either riboflavin or anthraquinone-2,6-disulfonate and these potential electron shuttles did not stimulate parental strain sulfate reduction with Fe0 as the electron donor. The results demonstrate that D. vulgaris primarily accepts electrons from Fe0 via H2 as an intermediary electron carrier. These findings clarify the interpretation of previous D. vulgaris corrosion studies and suggest that H2-mediated electron transfer is an important mechanism for iron corrosion under sulfate-reducing conditions.

Importance Microbial corrosion of iron in the presence of sulfate-reducing microorganisms is economically significant. There is substantial debate over how microbes accelerate iron corrosion. Tools for genetic manipulation have only been developed for a few Fe(III)-reducing and methanogenic microorganisms known to corrode iron and in each case those microbes were found to accept electrons from Fe0 via direct electron transfer. However, iron corrosion is often most intense in the presence of sulfate-reducing microbes. The finding that Desulfovibrio vulgaris relies on H2 to shuttle electrons between Fe0 and cells revives the concept, developed in some of the earliest studies on microbial corrosion, that sulfate reducers consumption of H2 is a major microbial corrosion mechanism. The results further emphasize that direct Fe0-to-microbe electron transfer has yet to be rigorously demonstrated in sulfate-reducing microbes.

Introduction

Microbial corrosion of iron-containing metals is a substantial economic problem, but the mechanisms are poorly understood (1–3). Sulfate reducers are often implicated in iron corrosion (1, 4–6). Desulfovibrio vulgaris has been the most studied pure culture isolate for investigating iron corrosion under sulfate-reducing conditions (7), dating back to the some of the earliest studies on microbial corrosion (8). Yet there is still substantial debate over how D. vulgaris corrodes iron. At least six mechanisms have been proposed (Figure 1).

Figure 1.
Figure 1.

Previously proposed mechanisms for Desulfovibrio vulgaris to promote the oxidation of Fe0. (A) Consuming H2 produced from Fe0 abiotically reducing H+. (B) Direct Fe0-to-microbe electron transfer. (C) H2S produced from sulfate reduction reacting with Fe0 to produce H2. (D) Flavin serving as an electron shuttle to promote electron transfer from Fe0 to cells. (E) Hydrogenases released from cells catalyzing Fe0 oxidation via reduction of H+ to H2. (F) Iron sulfide functioning as a conductive conduit for improved electron transfer to H+ for abiotic H2 production.

The first mechanism proposed (8) was abiotic oxidation of Fe0 coupled to proton reduction to generate H2: Embedded Image and consumption of the H2 produced via sulfate reduction: Embedded Image

The H2 uptake might stimulate H2 production by lowering H2 concentrations, thus making H2 generation more thermodynamically favorable.

In a second mechanism, hydrogenases released from moribund cells might accelerate reaction #1 by catalyzing H2 production from Fe0 (9). A third proposed mechanism is that the H2S generated from D. vulgaris sulfate reduction might react with Fe0 to generate H2 (1): Embedded Image

Alternatively Fe+2 and H2S can form iron sulfide precipitates that facilitate electron transfer from the Fe0 to H+, accelerating reaction #1 (2, 10).

Faster rates of corrosion following the addition of riboflavin (11–13) led to the suggestion that riboflavin can function as an electron shuttle that Fe0 reduces: Embedded Image and D. vulgaris oxidizes the reduced riboflavin with the reduction of sulfate: Embedded Image

However, those studies did not determine whether Fe0 could donate electrons to riboflavin or whether reduced riboflavin can serve as an electron donor for sulfate reduction. The alternative possibility that riboflavin might stimulate other aspects of microbial metabolism was also not evaluated.

Direct Fe0-to-microbe electron transfer for D. vulgaris has also been proposed (14). Studies with Geobacter (15, 16), Shewanella (17, 18), and Methanosarcina (19) species have provided evidence for direct electron uptake from Fe0 by: 1) eliminating the possibility that H2 was serving as an electron shuttle between Fe0 and cells; and 2) demonstrating with gene deletions that outer-surface c-type cytochromes were required for electron uptake from Fe0. In contrast, no studies have previously reported on D. vulgaris corrosion with strains that were unable to use H2 (7). Furthermore, D. vulgaris lacks outer-surface cytochromes (20) and no other D. vulgaris outer surface electrical contacts are known. Unlike the microbes previously shown to directly accept electrons from Fe0 (15–19), D. vulgaris does not directly reduce Fe(III) (21), a capability common to most electroactive microbes (22).

The suggestion that D. vulgaris might be capable of direct Fe0-to-microbe electron transfer is related to an earlier suggestion that D. ferrophilus has this capability (23). However, direct Fe0-to-microbe electron transfer by D. ferrophilus was only inferred (23). D. ferrophilus uses H2 as an electron donor and the possibility of H2 serving as intermediary electron carrier between Fe0 and D. ferrophilus was not rigorously ruled out those early studies (3). In subsequent studies, D. ferrophilus grew with pure Fe0 as the electron donor, but not with stainless steel (24). This distinction is important because pure Fe0 abiotically generates H2 via reaction #1 (15, 25), but stainless steel does not (16). In contrast to D. ferrophilus, stainless steel was an effective electron donor for Geobacter and Methanosarcina strains capable of direct electron uptake from Fe0 (16, 19, 24). Notably, protease digestion of D. ferrophilus extracellular proteins did not affect sulfate reduction rates with Fe0 as the electron donor (26), a result inconsistent with a microbe making direct electrical contact with Fe0. Therefore, the evidence available to date suggests that D. ferrophilus is most likely to accept electrons from Fe0 via a H2 intermediate (24).

A rigorous strategy to evaluate the possibility of H2 serving as an intermediary electron carrier is to determine whether strains unable to use H2 can respire with Fe0 as the sole electron donor (15, 16, 18, 19, 27). In instances in which the wild-type strain of interest can consume H2, this can be accomplished by deleting genes necessary for H2 uptake (15, 18, 27). A strain of D. vulgaris in which genes for all of the annotated hydrogenases on the genome were deleted is available as one of a large collection of mutant strains (28). We report here on studies on Fe0-dependent sulfate reduction conducted with this hydrogenase-deficient strain.

Results and Discussion

Hydrogenase mutant unable to grow with H2 as electron donor

The hydrogenase mutant grew as well as the parental strain in medium with lactate as the electron donor and sulfate as the electron acceptor (Figure 2A), but unlike the parental strain, the hydrogenase mutant did not grow in medium with H2 as the sole electron donor (Figure 2B). These results suggested that the hydrogenase mutant was a suitable strain to evaluate the role of H2 as an intermediary electron carrier during growth with Fe0 as the electron donor.

Figure 2.
Figure 2.

Growth of Desulfovibrio vulgaris parental strain and hydrogenase mutant with sulfate as the electron acceptor and either lactate (A) or H2 (B) as the electron donor, as measured by culture turbidity. Data are means and standard deviations of triplicate incubations.

Hydrogenase mutant cannot reduce sulfate with Fe0 as electron donor

The parental strain reduced sulfate with Fe0 as the sole electron donor, but the hydrogenase mutant did not (Figure 3A). The slight decline in sulfate over time in cultures with the hydrogenase mutant could be attributed to carry over of lactate with the inoculum because the final sulfate levels for the hydrogenase mutant with Fe0 were the same as for the parental strain without Fe0 (Figure 3A). As expected from previous studies under similar conditions (15), H2 accumulated in sterile controls (Figure 3B) reflecting abiotic Fe0 oxidation coupled to H+reduction. H2 also accumulated in cultures inoculated with the hydrogenase mutant, further demonstrating the inability of this strain to consume H2. H2 accumulated more in the hydrogenase mutant cultures than in the uninoculated control, probably due to the sulfide that was transferred along with the inoculum (see sulfide effect on H2 production below). In contrast, the parental strain maintained low H2 concentrations (Figure 3B), as expected for a microbe that can consume H2 produced from Fe0 (15). In the presence of Fe0, sulfate reduction of the parental strain declined (Figure 3A) as H2 production plateaued in abiotic controls (Figure 3B), consistent with H2 serving as the electron donor for sulfate reduction. These results indicated that H2 produced from Fe0 was an important electron donor for sulfate reduction by the parental strain.

Figure 3.
Figure 3.

Sulfate reduction and H2 concentrations with iron as the electron donor in the presence of Desulfovibrio vulgaris parental strain or hydrogenase mutant. Sulfate loss (A) or H2 concentrations (B) over time with pure Fe0 as the sole electron donor in the presence of the D. vulgaris strains and in controls without cells or without Fe0. (C) Lack of sulfate depletion with 316L stainless steel as the potential electron donor for the parental strain and in controls without cells or without stainless steel. (D) Sulfate loss over time in parental strain cultures amended with riboflavin, anthraquinone-2,6-disulfonate (AQDS), or with no amendments, with Fe0 as the sole electron donor. The bar graph inset in (D) shows lack of sulfate loss in cultures of the hydrogenase mutant with Fe0 as the electron donor and amendments of riboflavin or AQDS. Data are means and standard deviations of triplicate incubations.

However, the quantity of H2 that accumulated in abiotic Fe0-only controls or in the presence of Fe0 and the hydrogenase mutant was not sufficient to account for the amount of sulfate that the parental strain reduced with Fe0 as the electron donor. For example, on day 14 the parental strain had reduced 3.4 mM sulfate (50% of the time 0 concentration of 6.8 mM), which would require 13.6 mM H2 (4:1 stoichiometry of H2 oxidized per sulfate reduced, reaction #2). Only ca. half that much H2 accumulated in the hydrogenase mutant cultures (Figure 3B). One possibility for this disparity is that because rapid H2 uptake by the parental strain maintained low H2 concentrations (Figure 3B), H2 production from Fe0 (reaction #1) was more thermodynamically favorable, possibly accelerating H2 generation over that in the hydrogenase mutant cultures in which H2 accumulated.

Sulfide stimulates H2 production from Fe0

Sulfide that the parental strain generated from sulfate reduction with Fe0 as the electron donor is also likely to have promoted H2 production (Figure 4). Parental strain sulfide production was evident from the intense black precipitates indicative of iron sulfides on the Fe0 (Figure 4A). In contrast, there was only a small amount of iron sulfide on the Fe0 of the hydrogenase mutant cultures, which could be attributed to sulfide transferred along with the inoculum (Figure 4A). Sulfide was added to sterile medium, generating black iron sulfide precipitates (Figure 4B), to assess the possible sulfide impact on H2 production. Adding sulfide stimulated H2 generation (Figure 4C). One potential source of more H2 was the reaction of sulfide with Fe0 (reaction #3) in which there is a 1:1 stoichiometry for sulfide reacted and H2 produced. However, within 300 h the addition of 1.25 mM sulfide produced 2.7 mmol/liter H2 (Figure 4c), more than twice that expected from reaction #3. This result suggested that, as previously proposed (2, 10), iron sulfide precipitates also facilitated electron transfer from Fe0 to H+ (reaction #1), leading to additional H2 formation. Addition of 10-fold more sulfide only increased H2 an additional ca. 2-fold (Figure 4C), further demonstrating a lack of defined stoichiometry between sulfide additions and H2 formation.

Figure 4.
Figure 4.

Iron sulfide accumulations in culture and impact of sulfide on abiotic H2 production. (A) Appearance of uninoculated Fe0-containing medium and cultures inoculated with either the hydrogenase mutant or parental strain after 14 days of incubation. (B) Appearance of sterile Fe0-containing medium amended with 1.25 or 12.5 mM final concentration of sodium sulfide. (C) Accumulation of H2 over time in sterile media with and without added sulfide. Data are means and standard deviations of quadruplicate incubations.

Culture supernatant does not stimulate H2 production

Hydrogenases released from some microbes can accelerate H+ reduction with Fe0 (27, 29, 30) and hydrogenase activity has been detected in supernatants of moribund D. vulgaris cultures (9). However, supernatants from D. vulgaris cultures grown either with H2 or Fe0 did not stimulate H2 production from Fe0 over that in abiotic controls.

Stainless steel studies further indicate the importance of H2 as electron carrier

The inability of the hydrogenase mutant to reduce sulfate with Fe0 as the electron donor contrasts with electroactive microbes such as Geobacter sulfurreducens (15) or Shewanella oneidensis (18), which continue to utilize Fe0 as an electron donor even after gene deletions have eliminated the capability for H2 uptake. Both G. sulfurreducens and S. oneidensis are capable of direct electron uptake as evidenced from an inhibition of Fe0-based respiration when genes for key outer-surface c-type cytochromes are deleted (15, 17, 18). Thus, the lack of sulfate reduction by the D. vulgaris hydrogenase mutant suggests that it is unlikely to support sulfate reduction with direct electron uptake from Fe0.

This conclusion was further supported with the results of studies in which stainless steel was provided as the electron donor. Unlike pure Fe0, H2 production from stainless steel is minimal (16). However, microbes capable of direct electron uptake from Fe0 can extract electrons from stainless steel to support anaerobic respiration (16, 17, 19). D. vulgaris did not reduce sulfate with stainless steel as the electron donor (Figure 3C).

Electron shuttles do not promote Fe0-dependent sulfate reduction

An alternative proposed electron transfer mechanism in Fe0 corrosion is that flavins shuttle electrons between Fe0 and D. vulgaris (Figure 1). An observed increase in Fe0 corrosion when riboflavin is added to D. vulgaris cultures has been offered as evidence for flavin shuttling (11–13). However, the riboflavin amendments were to complex medium in which lactate was provided as an electron donor in addition to Fe0. It was not demonstrated that the riboflavin additions increased rates of Fe0-dependent sulfate reduction. In order to examine the possibility of electron shuttles facilitating electron transfer between Fe0 and D. vulgaris, studies were conducted under defined conditions with Fe0 as the sole electron donor for sulfate reduction and either riboflavin or the known electron shuttle anthraquinone-2,6,-disulfonate (AQDS) (31, 32). Riboflavin or AQDS did not accelerate Fe0-dependent sulfate reduction in the parental strain and did not enable the hydrogenase mutant to reduce sulfate with Fe0 as the electron donor (Figure 3D). The mid-point potentials of AQDS (−184 mV) and riboflavin (−208 mV) are probably too positive for the reduced form of these molecules to support the reduction of sulfate to sulfide (midpoint potential −217 mV). Therefore, the enhanced D. vulgaris Fe0 corrosion with riboflavin amendments (11–13) is likely to represent an impact of riboflavin on some aspect of growth or metabolism other than enhancement of electron transfer from Fe0 via an electron shuttle.

D. vulgaris attaches to Fe0 electron donor

The turbidity of D. vulgaris growing on Fe0 was very low compared to the turbidity in H2-grown cultures when a comparable amount of sulfate had been reduced (Figure 5A). Confocal scanning laser microscopy revealed that cells attached to the Fe0 surface (Figure 5B,C). The attachment of cells places cells at the point of H2 production, which should be advantageous because it enables H2 uptake at the point of production where localized H2 concentrations are higher than in the bulk surrounding environment. Furthermore, localized conditions at the cell/Fe0 interface are likely to accelerate Fe0 oxidation (Figure 5D). For example, attached D. vulgaris oxidizing H2 can make Fe0 oxidation more thermodynamically favorable, both by removing a product of the reaction (H2) and resupplying a reactant (H+) near the Fe0 surface. Sulfide produced at the Fe0 surface can further accelerate H2 production.

Figure 5.
Figure 5.

Desulfovibrio vulgaris attached to Fe0 serving as sole electron donor. (A) Lack of cell turbidity of culture growing with Fe0 as the source of H2 versus culture turbidity in culture grown under an atmosphere of H2. (B,C) Confocal scanning laser microscopy images showing cells attached to the Fe0 surface. (D) Mechanisms by which attachment to Fe0 might accelerate H2 production.

A common practice in Fe0 corrosion studies has been to infer that corrosion rates faster than that observed from abiotic H2 generation are indicative of corrosion mechanisms other than H2 serving as an intermediary electron carrier between Fe0 and cells (3). However, the possibilities for attached H2-consuming cells to accelerate H2 production from Fe0 illustrate the limitations to that reasoning.

Implications

Understanding how D. vulgaris promotes Fe0 oxidation is important because it is the microbe that has been used to develop much of the existing mechanistic framework to describe how sulfate reducers corrode Fe0 (7). The results demonstrate that the primary mechanism for D. vulgaris to reduce sulfate with Fe0 as an electron donor is with H2 serving as an electron shuttle between Fe0 and the cells. Sulfate was not reduced in the absence of genes required for H2 uptake, even when previously proposed organic electron shuttles were added. All the microbes that have been previously shown to be capable of direct electron uptake from Fe0 have outer-surface c-type cytochromes known to be involved in extracellular electron exchange with other donors/acceptors (15–19). D. vulgaris lacks outer-surface c-type cytochromes (20). Direct electron uptake from extracellular electron donors by routes other than cytochromes is possible (33). For example, several methanogen species that lack outer-surface c-type cytochromes appear to directly accept electrons from Geobacter metallireducens (34–37). However, the results presented here demonstrate that D. vulgaris does not function as an electrotroph with Fe0 as the electron donor. If D. vulgaris is representative of the sulfate reducers most responsible for the corrosion of ferrous metals, then potent hydrogenase inhibitors might provide a targeted approach to mitigate iron corrosion.

However, microbes other than sulfate reducers are also likely to contribute to corrosion (3, 30, 38, 39). Elucidating the mechanisms by which a diversity of microbes accelerate corrosion is essential for understanding why corrosion takes place, predicting corrosion rates under various environmental conditions, and developing strategies for corrosion prevention. The studies reported here further demonstrate that construction of appropriate mutants is a powerful approach to distinguish between a complexity of potential corrosion mechanisms.

Methods and Materials

Microbial Strains

Desulfovibrio vulgaris strains JW710 and JW5095, which were constructed in the laboratory of Judy Wall, University of Minnesota (28, 40), were provided from a repository of D. vulgaris mutants by Valentine V. Trotter and Adam M. Deutschbauer of the Lawrence Berkeley Laboratory. Strain JW710, is as a platform strain for a markerless genetic exchange system in D. vulgaris (40). The upp gene encoding uracil phosphoribosyltransferase has been deleted, to enable utilization of the upp gene as a counterselectable marker (40). Strain JW5095 was constructed by markerless deletion of all the hydrogenases that have been described in the D. vulgaris genome: DVU1921-22, DVU2525-26, DVU1917-18, DVU1769-70, DVU0429-34, DVU2286-93, and DVU1771 (28).

Culture Conditions

Cultures were routinely grown anaerobically at 37 °C in 10 ml of medium in 28 ml anaerobic pressure tubes (Bellco, Inc.), under N2/CO2 (80:20) in a modification of the previously described NBAF medium (41), designated NB medium. Per liter of deionized water NB medium contains: 0.42 g of KH2PO4, 0.22 g of K2HPO4, 0.2 g of NH4Cl, 0.38 g of KCl, 0.36 g of NaCl, 0.04 g of CaCl2 · 2H2O, 0.1 g of MgSO4 · 7H2O, 1.8 g of NaHCO3, 0.5 g of Na2CO3,, 1.0 ml of 1 mM Na2SeO4, 15.0 ml of a vitamin solution (42), and 10.0 ml of NB trace mineral solution. The composition of the NB trace mineral solution per liter of deionized water is 2.14 g of nitriloacetic acid, 0.1 g of MnCl2 · 4H2O, 0.3 g of FeSO4 · 7H2O, 0.17 g of CoCl2 · 6H2O, 0.2 g of ZnSO4 · 7H2O, 0.03 g of CuCl2 · 2H2O, 0.005 g of AlK(SO4)2 · 12H2O, 0.005 g of H3BO3, 0.09 g of Na2MoO4, 0.11 g of NiSO4 · 6H2O, and 0.02 g of Na2WO4 · 2H2O. Cells were routinely grown with sodium DL-lactate as the electron donor (20 mM) and sodium sulfate (20 mM) as the electron acceptor. Growth was monitored by inserting culture tubes directly into a spectrophotometer and determining A600. Growth with H2 as the sole electron donor was evaluated with 5 mM sodium acetate as a carbon source and H2 (140 kPa) as the sole electron donor. Cultures were routinely repressurized with H2 to compensate for any H2 consumption.

To evaluate growth with Fe0 as the potential electron donor, cells were grown in NB medium with Fe0 granules (2 g;1 to 2 mm diameter; Thermo Scientific) as the sole electron donor, 5 mM sulfate as the electron acceptor, and 5 mM sodium acetate as a carbon source. When specified, 50 μM riboflavin or 50 μM anthraquinone-2,6-disulfonate were added from concentrated anaerobic stock solutions. For studies with 316L stainless steel as the potential electron donor for sulfate reduction, 5 stainless steel cubes (5 mm x 3 mm x 3 mm) replaced the pure Fe0. The stainless steel cubes were polished with sand paper and the pure Fe0 and stainless steel were presterilized with ethanol as previously described (24).

Impact of Added Sulfide or Culture Supernatant on H2 Production

A final concentration of either 1.25 mM or 12.5 mM sodium sulfide was added to sterile Fe0-containing medium to determine whether sulfide stimulated H2 production. Culture filtrates were prepared by filtering late log grown cultures (Fe0-grown or H2-grown) through a 0.2 μM PES filter in a Coy anaerobic glove bag (gas phase 7:20:73 H2/CO2/N2) into pressure tubes with 2 g of Fe0. Tubes were resealed and flushed with N2:CO2 (80:20) for 5 minutes. Controls were sterile NB medium.

Analytical Methods

For sulfate determinations, culture aliquots (0.1 ml) were anaerobically withdrawn with a syringe and needle, filtered (0.22 μm, PVDF), and analyzed with a Dionex ICS-1000 with an AS22 column and AG22 guard with an eluent of 4.1 mM sodium carbonate and 1 mM sodium bicarbonate at 1.2 ml/min. H2 concentrations in the headspace were monitored on an Agilent 6890 gas chromatograph fitted with a thermal conductivity detector. The column was a Supelco Carboxen 1010 plot capillary column (30 m x 0.53 mm) with N2 carrier gas and 0.5 ml injections. The oven temperature was 40°C and the inlet was splitless at 5.5 psi and 225°C, the detector had a makeup flow of 7 ml/min and temperature of 225°C.

Confocal Microscopy

For confocal microscopy, Fe0 was gently removed from the pressure tube, soaked in isotonic wash buffer for 10 minutes, drained, stained for 10 minutes (Live/Dead BacLight bacterial viability kit (Thermo Fisher) (1 ml staining with 3 μl of each stain per ml)), and destained for 10 minutes in isotonic wash buffer. Fe0 pieces were then mounted on petri plates with an antifade/glycerol mixture. Cells were visualized with a 100x objective on a Nikon A1R-SIMe confocal microscope with NIS-Elements software.

Acknowledgements

These studies were made possible by the public availability Desulfovibrio vulgaris strains JW710 and JW5095 made in the laboratory of Judy Wall, University of Missouri. Thomas R. Juba constructed strain JW5095. We thank Valentine Trotter and Adam Deutschbauer of the Berkeley National Laboratory for providing us with strains used in this study. Confocal microscopy was performed in the Light Microscopy Facility and Nikon Center of Excellence at the Institute for Applied Life Sciences, UMass Amherst.

References

  1. 1.↵
    Enning D, Garrelfs J. 2014. Corrosion of iron by sulfate-reducing bacteria: new views of an old problem. Applied and Environmental Microbiology 80:1226–1236.
    OpenUrlAbstract/FREE Full Text
  2. 2.↵
    Little B, Hinks J, Blackwood DJ. 2020. Microbially influenced corrosion: Towards an interdisciplinary perspective on mechanisms. International Biodeterioration and Biodegradation 154:105062.
    OpenUrl
  3. 3.↵
    Lekbach Y, Liu T, Li Y, Moradia M, Dou W, Xu D, Smith JA, Lovley DR. 2021. Microbial corrosion of metals-the corrosion microbiome. Advances in Microbial Physiology 78:317–390.
    OpenUrl
  4. 4.↵
    Paisse S, Ghiglione J-F, Marty F, Abbas B, Gueune H, Amaya JMS, Muyzer G, Quillet L. 2013. Sulfate-reducing bacteria inhabiting natural corrosion deposits from marine steel structures. Applied Microbiology Biotechnology 97:7493–7504.
    OpenUrl
  5. 5.
    Rajala P, Cheng D-Q, Rice SA, Lauro FM. 2022. Sulfate-dependant microbially induced corrosion of mild steel in the deep sea: a 10-year microbiome study. Microbiome 10:4.
    OpenUrl
  6. 6.↵
    Wakai S, Eno N, Miyanaga K, Mizukami H, Sunaba T, Miyano Y. 2022. Dynamics of microbial communities on the corrosion behavior of steel in freshwater environment. Npj Materials Degradation 6:45.
    OpenUrl
  7. 7.↵
    Ueki T, Lovley DR. 2022. Desulfovibrio vulgaris as a model microbe for the study of corrosion under sulfate-reducing conditions. mLife 1:13–20.
    OpenUrl
  8. 8.↵
    von Wolzogen Kühr CAH, van der Vlugt LS. 1934. The graphitization of cast iron as an electrobiochemical process in anaerobic soil. Water 18:147–165.
    OpenUrl
  9. 9.↵
    Chatelus C, Carrier P, Saignes P, Libert MF, Berlier Y, Lespinat PA, Fauque G, Legall J. 1987. Hydrogenase activity in aged, nonviable Desulfovibrio vulgaris cultures and its significance in anaerobic biocorrosion. App Environ Microbio 53:1708–1710.
    OpenUrl
  10. 10.↵
    King RA, Miller JDA. 1971. Corrosion by the sulphate-reducing bacteria. Nature 233:491–492.
    OpenUrlCrossRefPubMedWeb of Science
  11. 11.↵
    Li H, Xu D, Li Y, Feng H, Liu Z, Li X, Gu T, Yang K. 2015. Extracellular electron transfer is a bottleneck in the microbiologically influenced corrosion of C1018 Carbon steel by the biofilm of sulfate-reducing bacterium Desulfovibrio vulgaris. PLoS ONE 10:e0136183.
    OpenUrlCrossRef
  12. 12.
    Zhang P, Xu D, Li Y, Yang K, Gu T. 2015. Electron mediators accelerate the microbiologically influenced corrosion of 304 stainless steel by the Desulfovibrio vulgaris biofilm. Bioelectrochemistry 101:14–21.
    OpenUrlCrossRef
  13. 13.↵
    Wang D, Liu J, Jia R, Dou W, Kumseranee S, Punpruk S, Li X, Gu T. 2020. Distinguishing two different microbiologically influenced corrosion (MIC) mechanisms using an electron mediator and hydrogen evolution detection. Corrosion Science 177:108993.
    OpenUrl
  14. 14.↵
    Gu T, Wang D, Lekbach Y, Xu D. 2021. Extracellular electron transfer in microbial biocorrosion. Current Opinion in Electrochemistry 29:100763.
    OpenUrl
  15. 15.↵
    Tang H-Y, Holmes DE, Ueki T, Palacios PA, Lovley DR. 2019. Iron corrosion via direct metal-microbe electron transfer. mBio 10:e00303–19.
    OpenUrl
  16. 16.↵
    Tang H-Y, Yang C, Ueki T, Pittman CC, Xu D, Woodard TL, Holmes DE, Gu T, Wang F, Lovley DR. 2021. Stainless steel corrosion via direct iron-to-microbe electron transfer by Geobacter species. ISME J 15:3084–3093.
    OpenUrl
  17. 17.↵
    Zhou E, Li F, Zhang D, Xu D, Li Z, Jia R, Jin Y, Song H, Li H, Wang Q, Wang J, Li X, Gu T, Homborg AM, Mol JMC, Smith JA, Wang F, Lovley DR. 2022. Direct microbial electron uptake as a mechanism for stainless steel corrosion in aerobic environments. Water Research 219:118553.
    OpenUrl
  18. 18.↵
    Hernandez-Santana A, Suflita JM, Nanny MA. 2022. Shewanella oneidensis MR-1 accelerates the corrosion of carbon steel using multiple electron transfer mechanisms. International Biodeterioration and Biodegradation 173:105439.
    OpenUrl
  19. 19.↵
    Holmes DE, Tang H-Y, Woodard TL, Liang D, Zhou J, Liu X, Lovley DR. 2022. Cytochrome-mediated direct electron uptake from metallic iron by Methanosarcina acetivorans. mLife (in press).
  20. 20.↵
    Heidelberg JF, Seshadri R, Haveman SA, Hemme CL, Paulsen IT, Kolonay JF, Eisen JA, Ward N, Methe B, Brinkac LM, Daugherty SC, Deboy RT, Dodson RJ, Durkin AS, Madupu R, Nelson WC, Sullivan SA, Fouts D, Haft DH, Selengut J, Peterson JD, Davidsen TM, Zafar N, Zhou L, Radune D, Dimitrov G, Hance M, Tran K, Khouri H, Gill J, Utterback TR, Feldblyum TV, Wall JD, Voordouw G, Fraser CM. 2004. The genome sequence of the anaerobic, sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough. Nat Biotechnol 22:554–9.
    OpenUrlCrossRefPubMedWeb of Science
  21. 21.↵
    Lovley DR, Roden EE, Phillips EJP, Woodward JC. 1993. Enzymatic iron and uranium reduction by sulfate-reducing bacteria. Marine Geol 113:41–53.
    OpenUrl
  22. 22.↵
    Lovley DR, Holmes DE. 2022. Electromicrobiology: The ecophysiology of phylogenetically diverse electroactive microorganisms. Nature Reviews Microbiology 20:5–19.
    OpenUrl
  23. 23.↵
    Dinh HT, Kuever J, MacBmann M, Hassel AW, Stratmann M, Widdel F. 2004. Iron corrosion by novel anaerobic microorganisms. Nature 427:829–832.
    OpenUrlCrossRefPubMedWeb of Science
  24. 24.↵
    Liang D, Liu X, Woodard TL, Holmes DE, Smith JA, Nevin KP, Feng Y, Lovley DR. 2021. Extracellular electron exchange capabilities of Desulfovibrio ferrophilus and Desulfopila corrodens. Environmental Science and Technology 55:16195–16203.
    OpenUrl
  25. 25.↵
    Daniels L, Belay N, Rajagopal BS, Weimer PJ. 1987. Bacterial methanogensis and growth from CO2 with elemental irion as the sole source of electrons. Science 237:509–511.
    OpenUrlAbstract/FREE Full Text
  26. 26.↵
    McCully, A.L., Spormann AM. 2020. Direct cathodic electron uptake coupled to sulfate reduction by Desulfovibrio ferrophilus IS5 biofilms. Environmental Microbiology 22:4794–4807.
    OpenUrl
  27. 27.↵
    Deutzmann JS, Sahin M, Spormann AM. 2015. Extracellular enzymes facilitate electron uptake in biocorrosion and bioelectrosynthesis. mBio 6:e00496–15.
    OpenUrlPubMed
  28. 28.↵
    Wall JD, Zane GM, Juba TR, Kuehl JV, Ray J, Chhabra SR, Trotter VV, Shatsky M, De León KB, Keller KL, Bender KS, Butland G, Arkin AP, Deutschbauer AM. 2021. Deletion mutants, archived transposon library, and tagged protein constructs of the model sulfatereducing bacterium Desulfovibrio vulgaris Hildenborough. Microbiol Resour Announc 10:e00072–21.
    OpenUrl
  29. 29.↵
    Rouvre I, Basseguy R. 2016. Exacerbation of the mild steel corrosion process by direct electron transfer between [Fe-Fe]-hydrogenase and material surface. Corrosion Science 111:199–211.
    OpenUrl
  30. 30.↵
    Tsurumaru H, Ito N, Mori K, Wakai S, Uchiyama T, Iino T, Hosoyama A, Ataku H, Nishijima K, Mise M, Shimizu A, Harada T, Horikawa H, Ichikawa N, Sekigawa T, Jinno K, Tanikawa S, Yamazaki J, Sasaki K, Yamazaki S, Fujita N, Harayama S. 2018. An extracellular [NiFe] hydrogenase mediating iron corrosion is encoded in a genetically unstable genomic island in Methanococcus maripaludis. Scientific Reports 8:1–10.
    OpenUrl
  31. 31.↵
    Lovley DR, Coates JD, Blunt-Harris EL, Phillips EJP, Woodward JC. 1996. Humic substances as electron acceptors for microbial respiration. Nature 382:445–448.
    OpenUrlCrossRefWeb of Science
  32. 32.↵
    Lovley D, Fraga J, Coates J, Blunt-Harris E. 1999. Humics as an electron donor for anaerobic respiration. Environmental Microbiology 1:89–98.
    OpenUrlCrossRefPubMedWeb of Science
  33. 33.↵
    Lovley DR. 2022. Electrotrophy: Other microbial species, iron, and electrodes as electron donors for microbial respirations. Bioresource Technology 345:126553.
    OpenUrl
  34. 34.↵
    Rotaru A-E, Shrestha PM, Liu F, Nevin KP, Lovley DR. 2014. Direct interspecies electron transfer during syntrophic growth of Geobacter metallireducens and Methanosarcina barkeri on ethanol. Appl Environ Microbiol 80:4599–4605.
    OpenUrlAbstract/FREE Full Text
  35. 35.
    Rotaru A-E, Shrestha PM, Liu F, Shrestha M, Shrestha D, Embree M, Zengler K, Wardman C, Nevin KP, Lovley DR. 2014. A new model for electron flow during anaerobic digestion: direct interspecies electron transfer to Methanosaeta for the reduction of carbon dioxide to methane. Energy & Environ Sci 7:408–415.
    OpenUrl
  36. 36.
    Holmes DE, Rotaru A-E, Ueki T, Shrestha PM, Ferry JG, Lovley DR. 2018. Electron and proton flux for carbon dioxide reduction in Methanosarcina barkeri during direct interspecies electron transfer Front Microbiol 9:3109.
    OpenUrl
  37. 37.↵
    Holmes DE, Zhou J, Smith JA, Wang C, Liu X, Lovley DR. 2022. Different outer membrane c-type cytochromes are involved in direct interspecies electron transfer to Geobacter or Methanosarcina species. mLife 1:272–286.
    OpenUrl
  38. 38.↵
    Emerson D. 2019. The role of iron-oxidizing bacteria in biocorrosion: a review. Biofouling:DOI: 10.1080/08927014.2018.1526281.
    OpenUrlCrossRef
  39. 39.↵
    Lahme S, Mand J, Longwell J, Smith R, Enning D. 2021. Severe corrosion of carbon steel in oil field produced water can be linked to methanogenic archaea containing a special type of [NiFe] hydrogenase. Appl Environ Microbiol 87:e01819-20.
    OpenUrl
  40. 40.↵
    Keller KL, Bender KS, Wall JD. 2009. Development of a markerless genetic exchange system for Desulfovibrio vulgaris Hildenborough and its use in generating a strain with increased transformation efficiency. Appl Environ Microbiol 75:7682–7691.
    OpenUrlAbstract/FREE Full Text
  41. 41.↵
    Coppi MV, Leang C, Sandler SJ, Lovley DR. 2001. Development of a genetic system for Geobacter sulfurreducens. Appl Environ Microbiol 67:3180–3187.
    OpenUrlAbstract/FREE Full Text
  42. 42.↵
    Lovley DR, Greening RC, Ferry JG. 1984. Rapidly growing rumen methanogenic organism that synthesizes coenzyme M and has a high affinity for formate. Apl Environ Microbiol 48:81–87.
    OpenUrl
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Posted November 15, 2022.
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H2 is a Major Intermediate in Desulfovibrio vulgaris Corrosion of Iron
Trevor L. Woodard, Toshiyuki Ueki, Derek R. Lovley
bioRxiv 2022.11.15.516606; doi: https://doi.org/10.1101/2022.11.15.516606
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