[Show abstract][Hide abstract] ABSTRACT: Shewanella-containing microbial fuel cells (MFCs) typically use the fresh water wild-type strain Shewanella oneidensis MR-1 due to its metabolic diversity and facultative oxidant tolerance. However, S. oneidensis MR-1 is not capable of metabolizing polysaccharides for extracellular electron transfer. The applicability of Shewanella japonica (an agar-lytic Shewanella strain) for power applications was analyzed using a diverse array of carbon sources for current generation from MFCs, cellular physiological responses at an electrode surface, biofilm formation, and the presence of soluble extracellular mediators for electron transfer to carbon electrodes. Critically, air-exposed S. japonica utilizes biosynthesized extracellular mediators for electron transfer to carbon electrodes with sucrose as the sole carbon source.
[Show abstract][Hide abstract] ABSTRACT: The ability of the metal reducer Shewanella oneidensis MR-1 to generate electricity in microbial fuel cells (MFCs) depends on the activity of a predicted type IV prepilin peptidase; PilD. Analysis of an S. oneidensis MR-1 pilD mutant indicated that it was deficient in pili production (Msh and type IV) and type II secretion (T2S). The requirement for T2S in metal reduction has been previously identified, but the role of pili remains largely unexplored. To define the role of type IV or Msh pili in electron transfer, mutants that lack one or both pilus biogenesis systems were generated and analyzed; a mutant that lacked flagella was also constructed and tested. All mutants were able to reduce insoluble Fe(III) and to generate current in MFCs, in contrast to the T2S mutant that is deficient in both processes. Our results show that loss of metal reduction in a PilD mutant is due to a T2S deficiency, and therefore the absence of c cytochromes from the outer surface of MR-1 cells, and not the loss of pili or flagella. Furthermore, MR-1 mutants deficient in type IV pili or flagella generated more current than the wild type, even though extracellular riboflavin levels were similar in all strains. This enhanced current generating ability is in contrast to a mutant that lacks the outer membrane c cytochromes, MtrC and OmcA. This mutant generated significantly less current than the wild type in an MFC and was unable to reduce Fe(III). These results indicated that although nanofilaments and soluble mediators may play a role in electron transfer, surface exposure of outer membrane c cytochromes was the determining factor in extracellular electron transfer in S. oneidensis MR-1.
[Show abstract][Hide abstract] ABSTRACT: Shewanella oneidensis is a facultative anaerobe that uses more than 14 different terminal electron acceptors for respiration. These include metal oxides and hydroxyoxides, and toxic metals such as uranium and chromium. Mutants deficient in metal reduction were isolated using the mariner transposon derivative, minihimar RB1. These included mutants with transposon insertions in the prepilin peptidase and type II secretion system genes. All mutants were deficient in Fe(III) and Mn(IV) reduction, and exhibited slow growth when DMSO was used as the electron acceptor. The genome sequence of S. oneidensis contains one prepilin peptidase gene, pilD. A similar prepilin peptidase that may function in the processing of type II secretion prepilins was not found. Single and multiple chromosomal deletions of four putative type IV pilin genes did not affect Fe(III) and Mn(IV) reduction. These results indicate that PilD in S. oneidensis is responsible for processing both type IV and type II secretion prepilin proteins. Type IV pili do not appear to be required for Fe(III) and Mn(IV) reduction.
[Show abstract][Hide abstract] ABSTRACT: Changes in metabolism and cellular physiology of facultative anaerobes during oxygen exposure can be substantial, but little is known about how these changes connect with electrical current output from an operating microbial fuel cell (MFC). A high-throughput voltage based screening assay (VBSA) was used to correlate current output from a MFC containing Shewanella oneidensis MR-1 to carbon source (glucose or lactate) utilization, culture conditions, and biofilm coverage over 250 h. Lactate induced an immediate current response from S. oneidensis MR-1, with both air-exposed and anaerobic anodes throughout the duration of the experiments. Glucose was initially utilized for current output by MR-1 when cultured and maintained in the presence of air. However, after repeated additions of glucose, the current output from the MFC decreased substantially while viable planktonic cell counts and biofilm coverage remained constant suggesting that extracellular electron transfer pathways were being inhibited. Shewanella maintained under an anaerobic atmosphere did not utilize glucose consistent with literature precedents. Operation of the VBSA permitted data collection from nine simultaneous S. oneidensis MR-1 MFC experiments in which each experiment was able to demonstrate organic carbon source utilization and oxygen dependent biofilm formation on a carbon electrode. These data provide the first direct evidence of complex cellular responses to electron donor and oxygen tension by Shewanella in an operating MFC at select time points.
Biotechnology and Bioengineering 02/2009; 103(3):524-31. · 4.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A miniature-microbial fuel cell (mini-MFC, chamber volume: 1.2 mL) was used to monitor biofilm development from a pure culture of Shewanella oneidensis DSP10 on graphite felt (GF) under minimal nutrient conditions. ESEM evidence of biofilm formation on GF is supported by substantial power density (per device cross-section) from the mini-MFC when using an acellular minimal media anolyte (1500 mW/m2). These experiments demonstrate that power density per volume for a biofilm flow reactor MFC should be calculated using the anode chamber volume alone (250W/m3), rather than with the full anolyte volume. Two oxygen reduction cathodes (uncoated GF or a Pt/vulcanized carbon coating on GF) were also compared to a cathode using uncoated GF and a 50mM ferricyanide catholyte solution. The Pt/C-GF (2-4% Pt by mass) electrodes with liquid cultures of DSP10 produced one order of magnitude larger power density (150W/m3) than bare graphite felt (12W/m3) in this design. These advances are some of the required modifications to enable the mini-MFC to be used in real-time, long-term environmental power generating situations.
[Show abstract][Hide abstract] ABSTRACT: The use of proton exchange membranes (PEMs) in biological fuel cells limits the diversity of novel designs for increasing output power or enabling autonomous function in unique environments. Here we show that selected nanoporous polymer filters (nylon, cellulose, or polycarbonate) can be used effectively in place of PEMs in a miniature microbial fuel cell (mini-MFC, device cross-section 2 cm2), generating a power density of 16 W/m3 with an uncoated graphite felt oxygen reduction reaction (ORR) cathode. The incorporation of polycarbonate or nylon membranes into biological fuel cell designs produced comparable power and durability to Nafion-117 membranes. Also, high power densities for novel larger (5 cm3 anode volume, 0.6 W/m3) and smaller (0.025 cm3 projected geometric volume, average power density 10 W/m3) chamberless and pumpless microbial fuel cells were observed. As an additional benefit, the nanoporous membranes isolated the anode from invading natural bacteria, increasing the potential applications for MFCs beyond aquatic sediment environments. This work is a practical solution for decreasing the cost of biological fuel cells while incorporating new features for powering long-term autonomous devices.
Environmental Science and Technology 03/2007; 41(4):1444-9. · 5.48 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A miniature microbial fuel cell (mini-MFC) is described that utilizes an aerobic culture of Shewanella oneidensis DSP10 as the active electrochemical species in the anode chamber. We find that the maximum aerobic mini-MFC power without the addition of exogenous mediators was 0.40 mW, a 33% decrease when compared with an anaerobic DSP10 culture (0.6 mW) operating in the mini-MFC. This decrease is most likely due to the presence of dissolved oxygen in the anode chamber that scavenges electrons to form water, thereby reducing the number of electrons donated to the anode. Aerobic power and current density at maximum power using the true surface area of the anode (611 cm2) were calculated to be 6.5 mW m−2 and 13 mA m−2. The power density rises to 2.0 W m−2 and 330 W m−3 when calculated using the cross-sectional area and volume of the device (2 cm2, 1.2 cm3). The Coulombic efficiency was also reduced from 11 to 5% when using the aerobic versus anaerobic culture. Similar results were found when the external mediator anthraquinone-2,6-disulfonate (AQDS) was added to the aerobic culture, resulting in a maximum power of 0.54 mW, a 37% drop in power when compared to the anaerobic mediated system.
[Show abstract][Hide abstract] ABSTRACT: A miniature microbial fuel cell (mini-MFC) is described that demonstrates high output power per device cross-section (2.0 cm2) and volume (1.2 cm3). Shewanella oneidensis DSP10 in growth medium with lactate and buffered ferricyanide solutions were used as the anolyte and catholyte, respectively. Maximum power densities of 24 and 10 mW/m2 were measured using the true surface areas of reticulated vitreous carbon (RVC) and graphite felt (GF) electrodes without the addition of exogenous mediators in the anolyte. Current densities at maximum power were measured as 44 and 20 mA/m2 for RVC and GF, while short circuit current densities reached 32 mA/m2 for GF anodes and 100 mA/m2 for RVC. When the power density for GF was calculated using the cross sectional area of the device or the volume of the anode chamber, we found values (3 W/m2, 500 W/m3) similar to the maxima reported in the literature. The addition of electron mediators resulted in current and power increases of 30-100%. These power densities were surprisingly high considering a pure S. oneidensis culture was used. We found that the short diffusion lengths and high surface-area-to-chamber volume ratio utilized in the mini-MFC enhanced power density when compared to output from similar macroscopic MFCs.
Environmental Science and Technology 05/2006; 40(8):2629-34. · 5.48 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This minimization procedure is only necessary for samples that must be viewed in the hydrated state without prior drying. A more direct installation sequence may be used for initially dry samples. For some samples, it may be important to avoid dehydration, but the presence of additional water is not a problem. In such cases it is recommended that the sample be covered in excess water before using the optimized installation sequence.