Dan Sun

Integrated hydrogen production process from cellulose by combining dark fermentation, microbial fuel cells, and a microbial electrolysis cell

Hydrogen gas production from cellulose was investigated using an integrated hydrogen production process consisting of a dark fermentation reactor and microbial fuel cells (MFCs) as power sources for a microbial electrolysis cell (MEC). Two MFCs (each 25 mL) connected in series to an MEC (72 mL) produced a maximum of 0.43 V using fermentation effluent as a feed, achieving a hydrogen production rate from the MEC of 0.48 m(3) H(2)/m(3)/d (based on the MEC volume), and a yield of 33.2 mmol H(2)/g COD removed in the MEC. The overall hydrogen production for the integrated system (fermentation, MFC and MEC) was increased by 41% compared with fermentation alone to 14.3 mmol H(2)/g cellulose, with a total hydrogen production rate of 0.24 m(3) H(2)/m(3)/d and an overall energy recovery efficiency of 23% (based on cellulose removed) without the need for any external electrical energy input.

Examination of microbial fuel cell start-up times with domestic wastewater and additional amendments

Rapid startup of microbial fuel cells (MFCs) and other bioreactors is desirable when treating wastewaters. The startup time with unamended wastewater (118 h) was similar to that obtained by adding acetate or fumarate (110-115 h), and less than that with glucose (181 h) or Fe(III) (353 h). Initial current production took longer when phosphate buffer was added, with startup times increasing with concentration from 149 h (25 mM) to 251 h (50 mM) and 526 h (100 mM). Microbial communities that developed in the reactors contained Betaproteobacteria, Acetoanaerobium noterae, and Chlorobium sp. Anode biomass densities ranged from 200 to 600 μg/cm(2) for all amendments except Fe(Ш) (1650 μg/cm(2)). Wastewater produced 91 mW/m(2), with the other MFCs producing 50 mW/m(2) (fumarate) to 103mW/m(2) (Fe(III)) when amendments were removed. These experiments show that wastewater alone is sufficient to acclimate the reactor without the need for additional chemical amendments.

A rapid selection strategy for an anodophilic consortium for microbial fuel cells

A rapid selection method was developed to enrich for a stable and efficient anodophilic consortium (AC) for microbial fuel cells (MFCs). A biofilm sample from a microbial electrolysis cell was serially diluted up to 10(-9) in anaerobic phosphate buffer solution and incubated in an Fe(III)-acetate medium, and an Fe(III)-reducing AC was obtained for dilutions up to 10(-6). The activity of MFC inoculated with the enrichment AC was compared with those inoculated with original biofilm or activated sludge. The power densities and Coulombic efficiencies of the AC (226 mW/m(2), 34%) were higher than those of the original biofilm (209 mW/m(2), 23%) and activated sludge (192 mW/m(2), 19%). The start-up period of the AC (60 h) was also shorter than those obtained with the other inocula (biofilm, 95h; activated sludge, 300 h). This indicated that such a strategy is highly efficient for obtaining an anodophilic consortium for improving the performance of an MFC.

Syntrophic Interactions Improve Power Production in Formic Acid Fed MFCs Operated With Set Anode Potentials or Fixed Resistances

Formic acid is a highly energetic electron donor but it has previously resulted in low power densities in microbial fuel cells (MFCs). Three different set anode potentials (−0.30, −0.15, and +0.15 V; vs. a standard hydrogen electrode, SHE) were used to evaluate syntrophic interactions in bacterial communities for formic acid degradation relative to a non‐controlled, high resistance system (1,000 Ω external resistance). No current was generated at −0.30 V, suggesting a lack of direct formic acid oxidation (standard reduction potential: −0.40 V). More positive potentials that allowed for acetic acid utilization all produced current, with the best performance at −0.15 V. The anode community in the −0.15 V reactor, based on 16S rDNA clone libraries, was 58% Geobacter sulfurreducens and 17% Acetobacterium, with lower proportions of these genera found in the other two MFCs. Acetic acid was detected in all MFCs suggesting that current generation by G. sulfurreducens was dependent on acetic acid production by Acetobacterium. When all MFCs were subsequently operated at an external resistance for maximum power production (100 Ω for MFCs originally set at −0.15 and +0.15 V; 150 Ω for the control), they produced similar power densities and exhibited the same midpoint potential of −0.15 V in first derivative cyclic voltammetry scans. All of the mixed communities converged to similar proportions of the two predominant genera (ca. 52% G. sulfurreducens and 22% Acetobacterium). These results show that syntrophic interactions can be enhanced through setting certain anode potentials, and that long‐term performance produces stable and convergent communities. Biotechnol. Bioeng. 2012; 109:405–414.

Improving startup performance with carbon mesh anodes in separator electrode assembly microbial fuel cells

In a separator electrode assembly microbial fuel cell, oxygen crossover from the cathode inhibits current generation by exoelectrogenic bacteria, resulting in poor reactor startup and performance. To determine the best approach for improving startup performance, the effect of acclimation to a low set potential (-0.2V, versus standard hydrogen electrode) was compared to startup at a higher potential (+0.2 V) or no set potential, and inoculation with wastewater or pre-acclimated cultures. Anodes acclimated to -0.2 V produced the highest power of 1330±60 mW m(-2) for these different anode conditions, but unacclimated wastewater inocula produced inconsistent results despite the use of this set potential. By inoculating reactors with transferred cell suspensions, however, startup time was reduced and high power was consistently produced. These results show that pre-acclimation at -0.2 V consistently improves power production compared to use of a more positive potential or the lack of a set potential.

Temporal-Spatial Changes in Viabilities and Electrochemical Properties of Anode Biofilms

Sustained current generation by anodic biofilms is a key element for the longevity and success of bioelectrochemical systems. Over time, however, inactive or dead cells can accumulate within the anode biofilm, which can be particularly detrimental to current generation. Mixed and pure culture (Geobacter anodireducens) biofilms were examined here relative to changes in electrochemical properties over time. An analysis of the three-dimensional metabolic structure of the biofilms over time showed that both types of biofilms developed a live outer-layer that covered a dead inner-core. This two-layer structure appeared to be mostly a result of relatively low anodic current densities compared to other studies. During biofilm development, the live layer reached a constant thickness, whereas dead cells continued to accumulate near the electrode surface. This result indicated that only the live outer-layer of biofilm was responsible for current generation and suggested that the dead inner-layer continued to function as an electrically conductive matrix. Analysis of the electrochemical properties and biofilm thickness revealed that the diffusion resistance measured using electrochemical impedance spectroscopy might not be due to acetate or proton diffusion limitations to the live layer, but rather electron-mediator diffusion.

Geobacter anodireducens sp. nov., an exoelectrogenic microbe in bioelectrochemical systems

A previously isolated exoelectrogenic bacterium, strain SD-1(T), was further characterized and identified as a representative of a novel species of the genus Geobacter. Strain SD-1(T) was Gram-negative, aerotolerant, anaerobic, non-spore-forming, non-fermentative and non-motile. Cells were short, curved rods (0.8-1.3 µm long and 0.3 µm in diameter). Growth of strain SD-1(T) was observed at 15-42 °C and pH 6.0-8.5, with optimal growth at 30-35 °C and pH 7. Analysis of 16S rRNA gene sequences indicated that the isolate was a member of the genus Geobacter, with the closest known relative being Geobacter sulfurreducens PCA(T) (98% similarity). Similar to other members of the genus Geobacter, strain SD-1(T) used soluble or insoluble Fe(III) as the sole electron acceptor coupled with the oxidation of acetate. However, SD-1(T) could not reduce fumarate as an electron acceptor with acetate oxidization, which is an important physiological trait for G. sulfurreducens. Moreover, SD-1(T) could grow in media containing as much as 3% NaCl, while G. sulfurreducens PCA(T) can tolerate just half this concentration, and this difference in salt tolerance was even more obvious when cultivated in bioelectrochemical systems. DNA-DNA hybridization analysis of strain SD-1(T) and its closest relative, G. sulfurreducens ATCC 51573(T), showed a relatedness of 61.6%. The DNA G+C content of strain SD-1(T) was 58.9 mol%. Thus, on the basis of these characteristics, strain SD-1(T) was not assigned to G. sulfurreducens, and was instead classified in the genus Geobacter as a representative of a novel species. The name Geobacter anodireducens sp. nov. is proposed, with the type strain SD-1(T) ( = CGMCC 1.12536(T) = KCTC 4672(T)).

Characterization of microbial communities during anode biofilm reformation in a two-chambered microbial electrolysis cell (MEC).

GeoChip (II) and single strand conformation polymorphism (SSCP) were used to characterize anode microbial communities of a microbial electrolysis cell (MEC). Biofilm communities, enriched in a two-chamber MEC (R1, 0.6 V applied) having a coulombic efficiency (CE) of 35±4% and a hydrogen yield (Y(H₂))of 31±3%, were used as the inoculum for a new reactor (R2). After three months R2 achieved stable performance with CE=38±4% and (Y(H₂)). Few changes in the predominant populations were observed from R1 to R2. Unlike sludge inoculation process in R1 in the beginning, little further elimination was aroused by community competitions in anode biofilm reformation in R2. Functional genes detection of biofilm indicated that cytochrome genes enriched soon in new reactor R2, and four genera (Desulfovibrio, Rhodopseudomonas, Shewanella and Geobacter) were likely to contribute to exoelectrogenic activity. This work also implied that symbiosis of microbial communities (exoelectrogens and others) contribute to system performance and stability.

Geobacter sp. SD-1 with enhanced electrochemical activity in high-salt concentration solutions

An isolate, designated strain SD-1, was obtained from a biofilm dominated by Geobacter sulfurreducens in a microbial fuel cell. The electrochemical activity of strain SD-1 was compared with type strains, G. sulfurreducens PCA and Geobacter metallireducens GS-15, and a mixed culture in microbial electrolysis cells. SD-1 produced a maximum current density of 290 ± 29 A m−3 in a high-concentration phosphate buffer solution (PBS-H, 200 mM). This current density was significantly higher than that produced by the mixed culture (189 ± 44 A m−3) or the type strains (< 70 A m−3). In a highly saline water (SW; 50 mM PBS and 650 mM NaCl), current by SD-1 (158 ± 4 A m−3) was reduced by 28% compared with 50 mM PBS (220 ± 4 A m−3), but it was still higher than that of the mixed culture (147 ± 19 A m−3), and strains PCA and GS-15 did not produce any current. Electrochemical tests showed that the improved performance of SD-1 was due to its lower charge transfer resistance and more negative potentials produced at higher current densities. These results show that the electrochemical activity of SD-1 was significantly different than other Geobacter strains and mixed cultures in terms of its salt tolerance.

Complete Genome Sequence of Geobacter anodireducens SD-1T, a Salt-Tolerant Exoelectrogenic Microbe in Bioelectrochemical Systems

ABSTRACT Strain SD-1 is the type strain of the species Geobacter anodireducens, which was originally isolated from a microbial fuel cell reactor in the United States. The characteristic of this bacterium is its high electrochemical activity. Here, we report the fully assembled genome and plasmid sequence of G. anodireducens SD-1T.