Cytochrome gene expression shifts in Geobacter sulfurreducens to maximize energy conservation in response to changes in redox conditions.

Previous studies have identified that Geobacter sulfurreducens has three different electron transfer pathways for respiration, and it switches between these pathways to adapt to the redox potential of its electron acceptor. However, only a small fraction of the electron carriers from each pathway have been identified. In this study, we combined electrochemical and gene expression data to identify electron carriers in the inner membrane, periplasm, outer membrane, and exterior of the cell that may be induced by the use of the three different electron transfer pathways. Cyclic voltammetry was performed on thin biofilms grown on anodes poised at different redox potentials, providing a quantitative assessment of the relative use of three electron-transfer pathways in each condition (catalytic midpoint potentials (EKAs) of -0.227 V [Low], -0.15 V [Medium], -0.1 V [High] vs. SHE). Transcriptomic analyses as a function of electrochemical signals or fumarate utilization showed differential induction in inner membrane (Medium: cbcL), periplasmic (Low: ppcB/ppcE, Medium: ppcA), outer membrane (Low: extA/extC, Medium: extJ/extK, Fumarate: extF/extG), and extracellular (Medium: omcZ, High/Fumarate: omcS/omcT) cytochromes, suggesting the pathway signals are associated with complex transcriptomic responses in genes across the electron transfer pathway. Our method combining electrochemical modeling and transcriptomics could be adapted to better understand electron transport in other electroactive organisms with complex metabolisms.

Intracytoplasmic membranes develop in Geobacter sulfurreducens under thermodynamically limiting conditions.

Geobacter sulfurreducens is an electroactive bacterium capable of reducing metal oxides in the environment and electrodes in engineered systems1,2. Geobacter sp. are the keystone organisms in electrogenic biofilms, as their respiration consumes fermentation products produced by other organisms and reduces a terminal electron acceptor e.g. iron oxide or an electrode. To respire extracellular electron acceptors with a wide range of redox potentials, G. sulfurreducens has a complex network of respiratory proteins, many of which are membrane-bound3-5. We have identified intracytoplasmic membrane (ICM) structures in G. sulfurreducens. This ICM is an invagination of the inner membrane that has folded and organized by an unknown mechanism, often but not always located near the tip of a cell. Using confocal microscopy, we can identify that at least half of the cells contain an ICM when grown on low potential anode surfaces, whereas cells grown at higher potential anode surfaces or using fumarate as electron acceptor had significantly lower ICM frequency. 3D models developed from cryo-electron tomograms show the ICM to be a continuous extension of the inner membrane in contact with the cytoplasmic and periplasmic space. The differential abundance of ICM in cells grown under different thermodynamic conditions supports the hypothesis that it is an adaptation to limited energy availability, as an increase in membrane-bound respiratory proteins could increase electron flux. Thus, the ICM provides extra inner-membrane surface to increase the abundance of these proteins. G. sulfurreducens is the first Thermodesulfobacterium or metal-oxide reducer found to produce ICMs.

Geobacter sulfurreducens' Unique Metabolism Results in Cells with a High Iron and Lipid Content.

Geobacter sulfurreducens is a ubiquitous iron-reducing bacterium in soils, and in engineered systems, it can respire an electrode to produce measurable electric current. Its unique metabolism, heavily dependent on an extensive network of cytochromes, requires a unique cell composition. In this work, we used metallomics, cell fraction and elemental analyses, and transcriptomics to study and analyze the cell composition of G. sulfurreducens. Elemental composition studies (C, H, O, N, and ash content) showed high C:O and H:O ratios of approximately 1.7:1 and 0.25:1, indicative of more reduced cell composition that is consistent with high lipid content. Our study shows that G. sulfurreducens cells have a large amount of iron (2 ± 0.2 µg/g dry weight) and lipids (32 ± 0.5% dry weight/dry weight) and that this composition does not change whether the cells are grown with a soluble or an insoluble electron acceptor. The high iron concentration, higher than similar microorganisms, is attributed to the production of cytochromes that are abundant in transcriptomic analyses in both solid and soluble electron acceptor growth. The unique cell composition of G. sulfurreducens must be considered when growing this microorganism for lab studies and commercial applications. IMPORTANCE Geobacter sulfurreducens is an electroactive microorganism. In nature, it grows on metallic minerals by transferring electrons to them, effectively "breathing" metals. In a manmade system, it respires an electrode to produce an electric current. It has become a model organism for the study of electroactive organisms. There are potential biotechnological applications of an organism that can bridge the gap between biology and electrical signal and, as a ubiquitous iron reducer in soils around the world, G. sulfurreducens has an impact on the global iron cycle. We measured the concentrations of metals, macromolecules, and basic elements in G. sulfurreducens to define this organism's composition. We also used gene expression data to discuss which proteins those metals could be associated with. We found that G. sulfurreducens has a large amount of lipid and iron compared to other bacteria-these observations are important for future microbiologists and biotechnologists working with the organism.

Organic carbon metabolism is a main determinant of hydrogen demand and dynamics in anaerobic soils.

Hydrogen (H2) is a crucial electron donor for many processes in the environment including nitrate-, sulfate- and, iron-reduction, homoacetogenesis, and methanogenesis, and is a major determinant of microbial competition and metabolic pathways in groundwater, sediments, and soils. Despite the importance of H2 for many microbial processes in the environment, the total H2 consuming capacity (or H2 demand) of soils is generally unknown. Using soil microcosms with added H2, the aims of this study were 1) to measure the H2 demand of geochemically diverse soils and 2) to define the processes leading to this demand. Study results documented a large range of H2 demand in soil (0.034-1.2 millielectron equivalents H2 g-1 soil). The measured H2 demand greatly exceeded the theoretical demand predicted based on measured concentrations of common electron acceptors initially present in a library of 15 soils. While methanogenesis accounted for the largest fraction of H2 demand, humic acid reduction and acetogenesis were also significant contributing H2-consuming processes. Much of the H2 demand could be attributed to CO2 produced during incubation from fermentation and/or acetoclastic methanogenesis. The soil initial total organic carbon showed the strongest correlation to H2 demand. Besides external additions, H2 was likely generated or cycled in the microcosms. Apart from fermentative H2 production, carboxylate elongation to produce C4-C7 fatty acids may have accounted for additional H2 production in these soils. Many of these processes, especially the organic carbon contribution is underestimated in microbial models for H2 consumption in natural soil ecosystems or during bioremediation of contaminants in soils.

Electrochemically Driven Photosynthetic Electron Transport in Cyanobacteria Lacking Photosystem II.

Light-activated photosystem II (PSII) carries out the critical step of splitting water in photosynthesis. However, PSII is susceptible to light-induced damage. Here, results are presented from a novel microbial electro-photosynthetic system (MEPS) that uses redox mediators in conjunction with an electrode to drive electron transport in live Synechocystis (ΔpsbB) cells lacking PSII. MEPS-generated, light-dependent current increased with light intensity up to 2050 µmol photons m-2 s-1, which yielded a delivery rate of 113 µmol electrons h-1 mg-chl-1 and an average current density of 150 A m-2 s-1 mg-chl-1. P700+ re-reduction kinetics demonstrated that initial rates exceeded wildtype PSII-driven electron delivery. The electron delivery occurs ahead of the cytochrome b6f complex to enable both NADPH and ATP production. This work demonstrates an electrochemical system that can drive photosynthetic electron transport, provides a platform for photosynthetic foundational studies, and has the potential for improving photosynthetic performance at high light intensities.

Determining global trends in syngas fermentation research through a bibliometric analysis.

Syngas fermentation, in which microorganisms convert H2, CO, and CO2 to acids and alcohols, is a promising alternative for carbon cycling and valorization. The intellectual landscape of the topic was characterized through a bibliometric analysis using a search query (SQ) that included all relevant documents on syngas fermentation available through the Web of Science database up to December 31st, 2021. The SQ was validated with a preliminary analysis in bibliometrix and a review of titles and abstracts of all sources. Although syngas fermentation began in the early 1980s, it grew rapidly beginning in 2008, with 92.5% of total publications and 87.3% of total citations from 2008 to 2021. The field has been steadily moving from fundamentals towards applications, suggesting that the field is maturing scientifically. The greatest number of publications and citations are from the USA, and researchers in China, Germany, and Spain also are highly active. Although collaborations have increased in the past few years, author-cluster analysis shows specialized research domains with little collaboration between groups. Based on topic trends, the main challenges to be address are related to mass-transfer limitations, and researchers are starting to explore mixed cultures, genetic engineering, microbial chain elongation, and biorefineries.

Carboxylates and alcohols production in an autotrophic hydrogen-based membrane biofilm reactor.

Microbiological conversion of CO2 into biofuels and/or organic industrial feedstock is an excellent carbon-cycling strategy. Here, autotrophic anaerobic bacteria in the membrane biofilm reactor (MBfR) transferred electrons from hydrogen gas (H2 ) to inorganic carbon (IC) and produced organic acids and alcohols. We systematically varied the H2 -delivery, the IC concentration, and the hydraulic retention time in the MBfR. The relative availability of H2 versus IC was the determining factor for enabling microbial chain elongation (MCE). When the H2 :IC mole ratio was high (>2.0 mol H2 /mol C), MCE was an important process, generating medium-chain carboxylates up to octanoate (C8, 9.1 ± 1.3 mM C and 28.1 ± 4.1 mmol C m-2 d-1 ). Conversely, products with two carbons were the only ones present when the H2 :IC ratio was low (<2.0 mol H2 /mol C), so that H2 was the limiting factor. The biofilm microbial community was enriched in phylotypes most similar to the well-known acetogen Acetobacterium for all conditions tested, but phylotypes closely related with families capable of MCE (e.g., Bacteroidales, Rhodocyclaceae, Alcaligenaceae, Thermoanaerobacteriales, and Erysipelotrichaceae) became important when the H2 :IC ratio was high. Thus, proper management of IC availability and H2 supply allowed control over community structure and function, reflected by the chain length of the carboxylates and alcohols produced in the MBfR.

Effect of pH on bacterial distributions within cathodic biofilm of the microbial fuel cell with maltodextrin as the substrate.

The aim of this study was to investigate pH effect on stratification of bacterial community in cathodic biofilm of the microbial fuel cell (MFC) under alkaline conditions. A single-chamber MFC with air-cathode was operated with 0.8 g/L maltodextrin and bicarbonate buffer solutions under pH values of 8.5, 9.5, and 10.5, respectively. The cathodic biofilms were characterized by linear sweep voltammetry (LSV), electrochemical impedance spectroscopy (EIS), confocal laser scanning microscopy (CLSM), freezing microtome and high-throughput sequencing analysis on bacterial communities, respectively. Results showed that the maximum power densities in the MFC increased with the pH values and reached 1221 ± 96 mW/m2 at pH = 10.5 during ∼30 d of operation. With different pH values, the composition and relative abundance of bacterial community significantly changed in the bottom (0-50 µm), middle (50-100 µm), and top (100-150 µm) layers of the cathodic biofilm. With pH = 10.5, aerobic bacteria accounted for 12%, 13%, and 34% of the bacterial community in the top, middle, and bottom layers, respectively. The amount of anaerobic bacteria in the top and middle layers (i.e., 52%, and 50% of the bacterial community, respectively) was higher than that in the bottom layer (22%). The distribution of aerobic and anaerobic bacteria showed a "valley-peak" structure within the layers. The high CO32- concentration facilitates the hydroxyl transfer and the neutralization in the anode of the MFC under high alkali conditions. The results from this study should be useful to develop new catalyst and cathode in the MFC.

Development, Fabrication, and Characterization of Composite Polycaprolactone Membranes Reinforced with TiO2 Nanoparticles.

This paper focuses on developing, fabricating, and characterizing composite polycaprolactone (PCL) membranes reinforced with titanium dioxide nanoparticles (NPs) elaborated by using two solvents; acetic acid and a mixture of chloroform and N,N-dimethylformamide (DMF). The resulting physical, chemical, and mechanical properties of the composite materials are studied by using experimental characterization techniques such as scanning electron microscopy (SEM), differential scanning calorimetry (DSC), X-Ray diffraction (XRD), Fourier-transform infrared (FTIR) analysis, contact angle (CA), uniaxial and biaxial tensile tests, and surface roughness measurements. Experimental results show that the composite material synthesized by sol-gel and chloroform-DMF has a better performance than the one obtained by using acetic acid as a solvent.

pH Dependency in Anode Biofilms of Thermincola ferriacetica Suggests a Proton-Dependent Electrochemical Response.

Monitoring the electrochemical response of anode respiring bacteria (ARB) helps elucidate the fundamental processes of anode respiration and their rate limitations. Understanding these limitations provides insights on how ARB create the complex interfacing of biochemical metabolic processes with insoluble electron acceptors and electronics. In this study, anode biofilms of the thermophilic (60 °C) Gram-positive ARB Thermincola ferriacetica were studied to determine the presence of a proton-dependent electron transfer response. The effects of pH, the presence of an electron donor (acetate), and biofilm growth were varied to determine their influence on the electrochemical midpoint potential ( EKA) and formal redox potential ( E°') under nonturnover conditions. The EKA and E°' are associated with an enzymatic process within ARB's metabolism that controls the rate and energetic state of their respiration. Results for all conditions indicate that pH was the major contributor to altering the energetics of T. ferriacetica anode biofilms. Electrochemical responses measured in the absence of an electron donor and with a minimal proton gradient within the anode biofilms resulted in a 48 ± 7 mV/pH unit shift in the E°', suggesting a proton-dependent rate-limiting process. Given the limited energy available for anode respiration (<200 mV when using acetate as electron donor), our results provide a new perspective in understanding proton-transport limitations in ARB biofilms, one in which ARB are thermodynamically limited by pH gradients. Since the anode biofilms of all ARB that perform direct extracellular electron transfer (EET) investigated thus far exhibit an n = 1 Nernstian behavior, and because this behavior is affected by changes in pH, we hypothesize that the Nernstian response is associated with membrane proteins responsible for proton translocation. Finally, this study shows that the EKA and E°' are a function of pH within the physiological range of ARB, and thus, given the significant effect pH has on this parameter, we recommend reporting the EKA and E°' of ARB biofilms at a specific bulk pH.

Geobacter Dominates the Inner Layers of a Stratified Biofilm on a Fluidized Anode During Brewery Wastewater Treatment.

In this study, we designed a microbial electrochemical fluidized bed reactor (ME-FBR), with an electroconductive anodic bed made of activated carbon particles for treating a brewery wastewater. Under a batch operating mode, acetate and propionate consumption rates were 13-fold and 2.4-fold higher, respectively, when the fluidized anode was polarized (0.2 V) with respect to open circuit conditions. Operating in a continuous mode, this system could effectively treat the brewery effluent at organic loading rates (OLR) over 1.7 kg m-3NRV d-1 and with removal efficiencies of 95 ± 1.4% (hydraulic retention time of 1 day and an influent of 1.7 g-COD L-1). The coulombic efficiency values highly depended upon the OLR applied, and varied from a 56 ± 15% to 10 ± 1%. Fluorescence in situ hybridization (FISH) analysis revealed a relative high abundance of Geobacter species (ca. 20%), and clearly showed a natural microbial stratification. Interestingly, the Geobacter cluster was highly enriched in the innermost layers of the biofilm (thickness of 10 µm), which were in contact with the electroconductive particles of bed, whereas the rest of bacteria were located in the outermost layers. To our knowledge, this is the first time that such a clear microbial stratification has been observed on an anode-respiring biofilm. Our results revealed the relevant role of Geobacter in switching between the electrode and other microbial communities performing metabolic reactions in the outermost environment of the biofilm.

Simultaneous fermentation of cellulose and current production with an enriched mixed culture of thermophilic bacteria in a microbial electrolysis cell.

An enriched mixed culture of thermophilic (60°C) bacteria was assembled for the purpose of using cellulose to produce current in thermophilic microbial electrolysis cells (MECs). Cellulose was fermented into sugars and acids before being consumed by anode-respiring bacteria (ARB) for current production. Current densities (j) were sustained at 6.5 ± 0.2 A m-2 in duplicate reactors with a coulombic efficiency (CE) of 84 ± 0.3%, a coulombic recovery (CR) of 54 ± 11% and without production of CH4 . Low-scan rate cyclic voltammetry (LSCV) revealed a mid-point potential (Eka ) of -0.17 V versus SHE. Pyrosequencing analysis of the V4 hypervariable region of 16S rDNA and scanning electron microscopy present an enriched thermophilic microbial community consisting mainly of the phylum Firmicutes with the Thermoanaerobacter (46 ± 13%) and Thermincola (28 ± 14%) genera occupying the biofilm anode in high relative abundance and Tepidmicrobium (38 ± 6%) and Moorella (11 ± 8%) genera present in high relative abundance in the bulk medium. The Thermoanaerobacter (15 ± 16%) and Brevibacillus (21 ± 30%) genera were also present in the bulk medium; however, their relative abundance varied by reactor. This study indicates that thermophilic consortia can obtain high CE and CR, while sustaining high current densities from cellulose in MECs.

Impact of carbon monoxide partial pressures on methanogenesis and medium chain fatty acids production during ethanol fermentation.

Medium-chain fatty acids (MCFA) are important biofuel precursors. Carbon monoxide (CO) is a sustainable electron and carbon donor for fatty acid elongation, since it is metabolized to MCFA precursors, it is toxic to most methanogens, and it is a waste product generated in the gasification of waste biomass. The main objective of this work was to determine if the inhibition of methanogenesis through the continuous addition of CO would lead to increased acetate or MCFA production during fermentation of ethanol. The effects of CO partial pressures (PCO ; 0.08-0.3 atm) on methanogenesis, fatty acids production, and the associated microbial communities were studied in batch cultures fed with CO and ethanol. Methanogenesis was partially inhibited at PCO ≥ 0.11 atm. This inhibition led to increased acetate production during the first phase of fermentation (0-19 days). However, a second addition of ethanol (day 19) triggered MCFA production only at PCO ≥ 0.11 atm, which probably occurred through the elongation of acetate with CO-derived ethanol and H2 :CO2 . Accordingly, during the second phase of fermentation (days 20-36), the distribution of electrons to acetate decreased at higher PCO , while electrons channeled to MCFA increased. Most probably, Acetobacterium, Clostridium, Pleomorphomonas, Oscillospira, and Blautia metabolized CO to H2 :CO2 , ethanol and/or fatty acids, while Peptostreptococcaceae, Lachnospiraceae, and other Clostridiales utilized these metabolites, along with the provided ethanol, for MCFA production. These results are important for biotechnological systems where fatty acids production are preferred over methanogenesis, such as in chain elongation systems and microbial fuel cells.

Changes in Glucose Fermentation Pathways as a Response to the Free Ammonia Concentration in Microbial Electrolysis Cells.

When a mixed-culture microbial electrolysis cell (MEC) is fed with a fermentable substrate, such as glucose, a significant fraction of the substrate's electrons ends up as methane (CH4) through hydrogenotrophic methanogenesis, an outcome that is undesired. Here, we show that free ammonia-nitrogen (FAN, which is NH3) altered the glucose fermentation pathways in batch MECs, minimizing the production of H2, the "fuel" for hydrogenotrophic methanogens. Consequently, the Coulombic efficiency (CE) increased: 57% for 0.02 g of FAN/L of fed-MEC, compared to 76% for 0.18 g of FAN/L of fed-MECs and 62% for 0.37 g of FAN/L of fed-MECs. Increasing the FAN concentration was associated with the accumulation of higher organic acids (e.g., lactate, iso-butyrate, and propionate), which was accompanied by increasing relative abundances of phylotypes that are most closely related to anode respiration (Geobacteraceae), lactic-acid production (Lactobacillales), and syntrophic acetate oxidation (Clostridiaceae). Thus, the microbial community established syntrophic relationships among glucose fermenters, acetogens, and anode-respiring bacteria (ARB). The archaeal population of the MEC fed 0.02 g FAN/L was dominated by Methanobacterium, but 0.18 and 0.37 g FAN/L led to Methanobrevibacter becoming the most abundant species. Our results provide insight into a way to decrease CH4 production and increase CE using FAN to control the fermentation step, instead of inhibiting methanogens using expensive or toxic chemical inhibitors, such as 2-bromoethanesulfonic acid.

H2O2 Production in Microbial Electrochemical Cells Fed with Primary Sludge.

We developed an energy-efficient, flat-plate, dual-chambered microbial peroxide producing cell (MPPC) as an anaerobic energy-conversion technology for converting primary sludge (PS) at the anode and producing hydrogen peroxide (H2O2) at the cathode. We operated the MPPC with a 9 day hydraulic retention time in the anode. A maximum H2O2 concentration of ∼230 mg/L was achieved in 6 h of batch cathode operation. This is the first demonstration of H2O2 production using PS in an MPPC, and the energy requirement for H2O2 production was low (∼0.87 kWh/kg H2O2) compared to previous studies using real wastewaters. The H2O2 gradually decayed with time due to the diffusion of H2O2-scavenging carbonate ions from the anode. We compared the anodic performance with a H2-producing microbial electrolysis cell (MEC). Both cells (MEC and MPPC) achieved ∼30% Coulombic recovery. While similar microbial communities were present in the anode suspension and anode biofilm for the two operating modes, aerobic bacteria were significant only on the side of the anode facing the membrane in the MPPC. Coupled with a lack of methane production in the MPPC, the presence of aerobic bacteria suggests that H2O2 diffusion to the anode side caused inhibition of methanogens, which led to the decrease in chemical oxygen demand removal. Thus, the Coulombic efficiency was ∼16% higher in the MPPC than in the MEC (64% versus 48%, respectively).

Electrochemical techniques reveal that total ammonium stress increases electron flow to anode respiration in mixed-species bacterial anode biofilms.

When anode-respiring bacteria (ARB) respire electrons to an anode in microbial electrochemical cells (MXCs), they harvest only a small amount of free energy. This means that ARB must have a high substrate-oxidation rate coupled with a high ratio of electrons used for respiration compared to total electrons removed by substrate utilization. It also means that they are especially susceptible to inhibition that slows anode respiration or lowers their biomass yield. Using several electrochemical techniques, we show that a relatively high total ammonium-nitrogen (TAN) concentration (2.2 g TAN/L) induced significant stress on the ARB biofilms, lowering their true yield and forcing the ARB to boost the ratio of electrons respired per electrons consumed from the substrate. In particular, a higher respiration rate, measured as current density (j), was associated with slower growth and a lower net yield, compared to an ARB biofilm grown with a lower ammonium concentration (0.2 g TAN/L). Further increases in influent TAN (to 3 and then to 4.4 g TAN/L) caused nearly complete inhibition of anode respiration. However, the ARB could recover from high-TAN inhibition after a shift of the MXC's feed to 0.2 g TAN/L. In summary, ARB biofilms were inhibited by a high TAN concentration, but could divert more electron flow toward anode respiration with modest inhibition and recover when severe inhibition was relieved. Biotechnol. Bioeng. 2017;114: 1151-1159. © 2017 Wiley Periodicals, Inc.

Tailoring Microbial Electrochemical Cells for Production of Hydrogen Peroxide at High Concentrations and Efficiencies.

A microbial peroxide producing cell (MPPC) for H2 O2 production at the cathode was systematically optimized with minimal energy input. First, the stability of H2 O2 was evaluated using different catholytes, membranes, and catalyst materials. On the basis of these results, a flat-plate MPPC fed continuously using 200 mm NaCl catholyte at a 4 h hydraulic retention time was designed and operated, producing H2 O2 for 18 days. H2 O2 concentration of 3.1 g L-1 H2 O2 with 1.1 Wh g-1 H2 O2 power input was achieved in the MPPC. The high H2 O2 concentration was a result of the optimum materials selected. The small energy input was largely the result of the 0.5 cm distance between the anode and cathode, which reduced ionic transport losses. However, >50 % of operational overpotentials were due to the 4.5-5 pH unit difference between the anode and cathode chambers. The results demonstrate that a MPPC can continuously produce H2 O2 at high concentration by selecting compatible materials and appropriate operating conditions.

Shifting the balance of fermentation products between hydrogen and volatile fatty acids: microbial community structure and function.

Fermentation is a key process in many anaerobic environments. Varying the concentration of electron donor fed to a fermenting community is known to shift the distribution of products between hydrogen, fatty acids and alcohols. Work to date has focused mainly on the fermentation of glucose, and how the microbial community structure is affected has not been explored. We fed ethanol, lactate, glucose, sucrose or molasses at 100 me- eq. L-1, 200 me- eq. L-1 or 400 me- eq. L-1 to batch-fed cultures with fermenting, methanogenic communities. In communities fed high concentrations of electron donor, the fraction of electrons channeled to methane decreased, from 34% to 6%, while the fraction of electrons channeled to short chain fatty acids increased, from 52% to 82%, averaged across all electron donors. Ethanol-fed cultures did not produce propionate, but did show an increase in electrons directed to acetate as initial ethanol concentration increased. In glucose, sucrose, molasses and lactate-fed cultures, propionate accumulation co-occurred with known propionate producing organisms. Overall, microbial communities were determined by the substrate provided, rather than its initial concentration, indicating that a change in community function, rather than community structure, is responsible for shifts in the fermentation products produced.

The effect of pH and buffer concentration on anode biofilms of Thermincola ferriacetica.

We assessed the effects of pH and buffer concentration on current production and growth of biofilms of Thermincola ferriacetica - a thermophilic, Gram-positive, anode-respiring bacterium (ARB) - grown on anodes poised at a potential of -0.06V vs. SHE in microbial electrolysis cells (MECs) at 60°C. T. ferriacetica generated current in the pH range of 5.2 to 8.3 with acetate as the electron donor and 50mM bicarbonate buffer. Maximum current density was reduced by ~80% at pH5.2 and ~14% at 7.0 compared to pH8.3. Increasing bicarbonate buffer concentrations from 10mM to 100mM resulted in an increase in the current density by 40±6%, from 6.8±1.1 to 11.2±2.7Am(-2), supporting that more buffer alleviated pH depression within T. ferriacetica biofilms. Confocal laser scanning microscopy (CLSM) images indicated that higher bicarbonate buffer concentrations resulted in larger live biofilm thicknesses: from 68±20µm at 10mM bicarbonate to >150µm at 100mM, supporting that buffer availability was a strong influence on biofilm thickness. In comparison to mesophilic Geobacter sulfurreducens biofilms, the faster transport rates at higher temperature and the ability to grow at relatively lower pH allowed T. ferriacetica to produce higher current densities with lower buffer concentrations.

Evaluating biochemical methane production from brewer's spent yeast.

Anaerobic digestion treatment of brewer's spent yeast (SY) is a viable option for bioenergy capture. The biochemical methane potential (BMP) assay was performed with three different samples (SY1, SY2, and SY3) and SY1 dilutions (75, 50, and 25 % on a v/v basis). Gompertz-equation parameters denoted slow degradability of SY1 with methane production rates of 14.59-4.63 mL/day and lag phases of 10.72-19.7 days. Performance and kinetic parameters were obtained with the Gompertz equation and the first-order hydrolysis model with SY2 and SY3 diluted 25 % and SY1 50 %. A SY2 25 % gave a 17 % of TCOD conversion to methane as well as shorter lag phase (<1 day). Average estimated hydrolysis constant for SY was 0.0141 (±0.003) day(-1), and SY2 25 % was more appropriate for faster methane production. Methane capture and biogas composition were dependent upon the SY source, and co-digestion (or dilution) can be advantageous.