Hyung Sool Lee

A kinetic perspective on extracellular electron transfer by anode-respiring bacteria.

In microbial fuel cells and electrolysis cells (MXCs), anode-respiring bacteria (ARB) oxidize organic substrates to produce electrical current. In order to develop an electrical current, ARB must transfer electrons to a solid anode through extracellular electron transfer (EET). ARB use various EET mechanisms to transfer electrons to the anode, including direct contact through outer-membrane proteins, diffusion of soluble electron shuttles, and electron transport through solid components of the extracellular biofilm matrix. In this review, we perform a novel kinetic analysis of each EET mechanism by analyzing the results available in the literature. Our goal is to evaluate how well each EET mechanism can produce a high current density (> 10 A m(-2)) without a large anode potential loss (less than a few hundred millivolts), which are feasibility goals of MXCs. Direct contact of ARB to the anode cannot achieve high current densities due to the limited number of cells that can come in direct contact with the anode. Slow diffusive flux of electron shuttles at commonly observed concentrations limits current generation and results in high potential losses, as has been observed experimentally. Only electron transport through a solid conductive matrix can explain observations of high current densities and low anode potential losses. Thus, a study of the biological components that create a solid conductive matrix is of critical importance for understanding the function of ARB.

Biological hydrogen production: prospects and challenges

Hydrogen gas provides exceptional value as an energy carrier and industrial feedstock, but currently is produced entirely by reforming fossil fuels. Biological hydrogen production (BioH(2)), which offers the possibility of being renewable and carbon neutral, can be achieved by photosynthesis, fermentation, and microbial electrolysis cells. This review introduces the principles, advantages and challenges of each approach to BioH(2). Photosynthetic BioH(2) is the ultimate renewable source, since it directly uses inexhaustible resources: sunlight energy and electrons from H(2)O. However, it presents major technical challenges, particularly due to oxygen sensitivity. Fermentative BioH(2) offers a high production rate, but poor conversion efficiency from the organic substrate to H(2). The microbial electrolysis cell can achieve high conversion efficiency, but is an emerging technology.

Syntrophic interactions among anode respiring bacteria (ARB) and Non‐ARB in a biofilm anode: electron balances

We demonstrate that the coulombic efficiency (CE) of a microbial electrolytic cell (MEC) fueled with a fermentable substrate, ethanol, depended on the interactions among anode respiring bacteria (ARB) and other groups of micro‐organisms, particularly fermenters and methanogens. When we allowed methanogenesis, we obtained a CE of 60%, and 26% of the electrons were lost as methane. The only methanogenic genus detected by quantitative real‐time PCR was the hydrogenotrophic genus, Methanobacteriales, which presumably consumed all the hydrogen produced during ethanol fermentation (∼30% of total electrons). We did not detect acetoclastic methanogenic genera, indicating that acetate‐oxidizing ARB out‐competed acetoclastic methanogens. Current production and methane formation increased in parallel, suggesting a syntrophic interaction between methanogens and acetate‐consuming ARB. When we inhibited methanogenesis with 50 mM 2‐bromoethane sulfonic acid (BES), the CE increased to 84%, and methane was not produced. With no methanogenesis, the electrons from hydrogen were converted to electrical current, either directly by the ARB or channeled to acetate through homo‐acetogenesis. This illustrates the key role of competition among the various H2 scavengers and that, when the hydrogen‐consuming methanogens were present, they out‐competed the other groups. These findings also demonstrate the importance of a three‐way syntrophic relationship among fermenters, acetate‐consuming ARB, and a H2 consumer during the utilization of a fermentable substrate. To obtain high coulombic efficiencies with fermentable substrates in a mixed population, methanogens must be suppressed to promote new interactions at the anode that ultimately channel the electrons from hydrogen to current. Biotechnol. Bioeng. 2009;103: 513–523.

Fate of H2 in an upflow single-chamber microbial electrolysis cell using a metal-catalyst-free cathode

With the goal of maximizing the H2-harvesting efficiency, we designed an upflow single-chamber microbial electrolysis cell (MEC) by placing the cathode on the top of the MEC and carried out a program to track the fate of H2 and electron equivalents in batch experiments. When the initial acetate concentration was 10 mM in batch-evaluation experiments lasting 32 h, the cathodic conversion efficiency (CCE) from coulombs (i.e., electron equivalents in current from the anode to the cathode) to H2 was 98 +/- 2%, the Coulombic efficiency (CE) was 60 +/- 1%, the H2 yield was 59 +/- 2%, and methane production was negligible. However, longer batch reaction time (approximately 7 days) associated with higher initial acetate concentrations (30 or 80 mM) led to significant H2 loss due to CH4 accumulation: up to 14 +/- 1% and 16 +/- 2% of the biogas at 30 and 80 mM of acetate, respectively. Quantitative PCR proved that no acetoclastic methanogens were present, but that hydrogenotrophic methanogens (i.e., Methanobacteriales) were present on both electrodes. The hydrogenotrophic methanogens decreased the CCE by diverting H2 generated at the cathode to CH4 in the upflow single-chamber MEC. In some experiments, the CE was greater than 100%. The cause was anode-respiring bacteria oxidizing H2 and producing current which recycled H2 between the cathode and the anodes, increasing CE to over 100%, but with a concomitant decline in CCE, despite negligible CH4 formation.

Significance of biological hydrogen oxidation in a continuous single-chamber microbial electrolysis cell.

A single-chamber microbial electrolysis cell (MEC) that used a high density of nonmetal-catalyst carbon fibers as the anode achieved high volumetric current densities from 1470 +/- 60 to 1630 +/- 50 A/m(3) for a hydraulic retention time of 1.6-6.5 h. The high current density was driven by a large anode surface area and corresponded to a volumetric chemical oxygen demand (COD)-removal rate of 27-49 kg COD/m(3).d. Observed H(2) harvesting rates were from 2.6 +/- 0.10 to 4.3 +/- 0.46 m(3) H(2)/m(3).d, but the H(2) production rates computed from the current densities were 16.3-18.2 m(3) H(2)/m(3).d. Tracking all significant electron sinks (residual acetate, H(2), CH(4), biomass, and soluble microbial products (SMP)) in the single-chamber MEC showed that H(2) reoxidation by anode-respiring bacteria recycled H(2) between the cathode and the anode, and this caused the large discrepancy in H(2) production and harvest rates. H(2) recycle accounted for 62-76% of observed current density, and this made the observed Coulombic efficiency 190-310% at steady state. Consequently, the cathodic conversion efficiency was only 16-24%. The current density added by H(2) recycle also increased the applied voltage from approximately 0.6 V to approximately 1.5 V for the highest H(2) harvest rate (4.3 m(3) H(2)/m(3).d). CH(4) generation consistently occurred in the continuous single-chamber MEC, and its electron fraction of consumed acetate was 7-25%. Because of methane formation and biomass/SMP accumulation, the overall H(2) recovery was moderate at 1.8-2.0 mol of H(2)/mol of acetate in the MEC. Thus, this study illustrates that a single-chamber MEC with a high anode surface area can generate high volumetric rates for COD removal and H(2) generation, but H(2) recycle and methanogenesis present significant challenges for practical application.

A μL-scale micromachined microbial fuel cell having high power density

We report a MEMS (Micro-Electro-Mechanical Systems)-based microbial fuel cell (MFC) that produces a high power density. The MFC features 4.5-μL anode/cathode chambers defined by 20-μm-thick photo-definable polydimethylsiloxane (PDMS) films. The MFC uses a Geobacter-enriched mixed bacterial culture, anode-respiring bacteria (ARB) that produces a conductive biofilm matrix. The MEMS MFC generated a maximum current density of 16,000 μA cm(-3) (33 μA cm(-2)) and power density of 2300 μW cm(-3) (4.7 μW cm(-2)), both of which are substantially greater than achieved by previous MEMS MFCs. The coulombic efficiency of the MEMS MFC was at least 31%, by far the highest value among reported MEMS MFCs. The performance improvements came from using highly efficient ARB, minimizing the impact of oxygen intrusion to the anode chamber, having a large specific surface area that led to low internal resistance.

Evaluation of metabolism using stoichiometry in fermentative biohydrogen

We first constructed full stoichiometry, including cell synthesis, for glucose mixed‐acid fermentation at different initial substrate concentrations (0.8–6 g‐glucose/L) and pH conditions (final pH 4.0–8.6), based on experimentally determined electron‐equivalent balances. The fermentative bioH2 reactions had good electron closure (−9.8 to +12.7% for variations in glucose concentration and −3 to +2% for variations in pH), and C, H, and O errors were below 1%. From the stoichiometry, we computed the ATP yield based on known fermentation pathways. Glucose‐variation tests (final pH 4.2–5.1) gave a consistent fermentation pattern of acetate + butyrate + large H2, while pH significantly shifted the catabolic pattern: acetate + butyrate + large H2 at final pH 4.0, acetate + ethanol + modest H2 at final pH 6.8, and acetate + lactate + trivial H2 at final pH 8.6. When lactate or propionate was a dominant soluble end product, the H2 yield was very low, which is in agreement with the theory that reduced ferredoxin (Fdred) formation is required for proton reduction to H2. Also consistent with this hypothesis is that high H2 production correlated with a high ratio of butyrate to acetate. Biomass was not a dominant sink for electron equivalents in H2 formation, but became significant (12%) for the lowest glucose concentration (i.e., the most oligotrophic condition). The fermenting bacteria conserved energy similarly at ∼3 mol ATP/mol glucose (except 0.8 g‐glucose/L, which had ∼3.5 mol ATP/mol glucose) over a wide range of H2 production. The observed biomass yield did not correlate with ATP conservation; low observed biomass yields probably were caused by accelerated rates of decay or production of soluble microbial products. Biotechnol. Bioeng. 2009; 102: 749–758.

A paper-based microbial fuel cell: instant battery for disposable diagnostic devices.

We present a microfabricated paper-based microbial fuel cell (MFC) generating a maximum power of 5.5 μW/cm(2). The MFC features (1) a paper-based proton exchange membrane by infiltrating sulfonated sodium polystyrene sulfonate and (2) micro-fabricated paper chambers by patterning hydrophobic barriers of photoresist. Once inoculum and catholyte were added to the MFC, a current of 74 μA was generated immediately. This paper-based MFC has the advantages of ease of use, low production cost, and high portability. The voltage produced was increased by 1.9 × when two MFC devices were stacked in series, while operating lifetime was significantly enhanced in parallel.

Hydrogen consumption in microbial electrochemical systems (MXCs): The role of homo-acetogenic bacteria

Homo-acetogens in the anode of a microbial electrolysis cell (MEC) fed with H(2) as sole electron donor allowed current densities similar to acetate-fed biofilm anodes (∼10 A/m(2)). Evidence for homo-acetogens included accumulation of acetate at high concentrations (up to 18 mM) in the anode compartment; detection of formate, a known intermediate during reductive acetogenesis by the acetyl-CoA pathway; and detection of formyl tetrahydrofolate synthetase (FTHFS) genes by quantitative real-time PCR. Current production and acetate accumulation increased in parallel in batch and continuous mode, while both values decreased simultaneously at short hydraulic retention times (1h) in the anode compartment, which limited suspended homo-acetogens. Acetate produced by homo-acetogens accounted for about 88% of the current density of 10A/m(2), but the current density was sustained at 4A/m(2) at short hydraulic retention time because of a robust partnership of homo-acetogens and anode respiring bacteria (ARB) in the biofilm anode.

Using a pulsed electric field as a pretreatment for improved biosolids digestion and methanogenesis

Researchers tested using pulsed electric field (PEF) technology to enhance conversion of organic solids material in waste activated sludge (WAS) and pig manure to soluble and colloidal forms, which are more bioavailable for methane production during subsequent anaerobic digestion. Operating parameters were varied from 1.1 to 19.8 kWhr/m3 to show the influence amount of treatment has on soluble chemical oxygen demand (SCOD), small colloidal solids, and methane production via the biochemical methane potential test. When PEF treatment exceeded a threshold, which was approximately 10 kWhr/m3, focused pulsed treatment solubilized approximately 10% of the total COD, increasing SCOD from as low as 20 mg/L to more than 1000 mg/L. The process also disrupted a larger portion of the volatile suspended solids (VSS) to form small colloids not measured by the VSS assay (between 0.2 and 1.2 microm). The effects increased the biological methane potential of the samples significantly: by 80% for pig manure and 100% for WAS after 25 to 30 days. These results support the conclusion that PEF pretreatment before anaerobic digestion has the potential to significantly improve digester performance, resulting in added methane production and decreased residual biosolids.