Project description:Microbial Fuel Cells (MFCs) are innovative environmental engineering systems that harness the metabolic activities of microbial communities to convert chemical energy in waste into electrical energy. However, MFC performance optimization remains challenging due to limited understanding of microbial metabolic mechanisms, particularly with complex substrates under realistic environmental conditions. This study investigated the effects of substrate complexity (acetate vs. starch) and varying mass transfer (stirred vs. non-stirred) on acclimatization rates, substrate degradation, and microbial community dynamics in air-cathode MFCs. Stirring was critical for acclimating to complex substrates, facilitating electrogenic biofilm formation in starch-fed MFCs, while non-stirred MFCs showed limited performance under these conditions. Non-stirred MFCs, however, outperformed stirred systems in current generation and coulombic efficiency (CE), especially with simple substrates (acetate), achieving 66% CE compared to 38% under stirred conditions, likely due to oxygen intrusion in the stirred systems. Starch-fed MFCs exhibited consistently low CE (19%) across all tested conditions due to electron diversion into volatile fatty acids (VFA). Microbial diversity was higher in acetate-fed MFCs but unaffected by stirring, while starch-fed MFCs developed smaller, more specialized communities. Kinetic analysis identified hydrolysis of complex substrates as the rate-limiting step, with rates an order of magnitude slower than acetate consumption. Combined hydrolysis-fermentation rates were unaffected by stirring, but stirring significantly impacted acetate consumption rates, likely due to oxygen-induced competition between facultative aerobes and electrogenic bacteria. These findings highlight the trade-offs between enhanced substrate availability and oxygen-driven competition in MFCs. For real-world applications, initiating reactors with dynamic stirring to accelerate acclimatization, followed by non-stirred operation, may optimize performance. Integrating MFCs with anaerobic digestion could overcome hydrolysis limitations, enhancing the degradation of complex substrates while improving energy recovery. This study introduces novel strategies to address key challenges in scaling up MFCs for wastewater treatment, bridging the gap between fundamental research and practical applications to advance environmental systems.
Project description:Electrochemically active bacteria (EAB) are capable of electrochemical interactions with electrodes via extracellular electron transfer (EET) pathways and serve as essential components in bioelectrochemical systems. Previous studies have suggested that EAB, such as Shewanella oneidensis MR-1, use cyclic AMP (cAMP) receptor proteins for coordinately regulating the expression of catabolic and EET-related genes, allowing us to hypothesize that the intracellular cAMP concentration is an important factor determining electrochemical activities of EAB. The present study constructed an MR-1 mutant, cyaC-OE that overexpressed cyaC, a gene encoding a membrane-bound class III adenylate cyclase, and examined its electrochemical and transcriptomic characteristics. We show that intracellular cAMP concentration in cyaC-OE is more than double that in wild-type MR-1, and cya-OE generates approximately two-fold higher current in BES than the wild type. In addition, the expression of genes involved in EET and anaerobic carbon catabolism is up-regulated in cya-OE as compared to that in the wild type. These results suggest that enhancement of the intracellular cAMP level is a promising approach for constructing an EAB with high catabolic and electrochemical activities.
Project description:We investigated ion transport limitations on 3D graphite felt electrodes by growing Geobacter sulfurreducens biofilms with advection to eliminate external mass transfer limitations. We characterized ion transport limitations by: (i) showing that serially increasing NaCl concentration up to 200 mM increased current linearly up to a total of +273% vs. 0 mM NaCl under advective conditions; (ii) growing the biofilm with a starting concentration of 200 mM NaCl, which led to a maximum current increase of 400% vs. current generation without NaCl, and (iii) showing that un-colonized surface area remained even after steady-state current was reached. After accounting for iR effects, we confirmed that the excess surface area existed despite a non-zero overpotential. The fact that the biofilm was constrained from colonizing and producing further current under these conditions confirmed the biofilms under study here were ion transport-limited. Our work demonstrates that the use of high surface area electrodes may not increase current density when the system design allows ion transport limitations to become dominant.
Project description:To get insights in the electrogenic anaerobic lifestyle of P. putida KT2440 cultivated in a bioelectrochemical system (BES), we employed whole genome microarray expression profile.
Project description:We report an integrated experimental and simulation study of ammonia recovery using microbial electrolysis cells (MECs). The transport of various species during the batch-mode operation of an MEC was examined experimentally and the results were used to validate the mathematical model for such an operation. It was found that, while the generated electrical current through the system tends to acidify (or basify) the anolyte (or catholyte), their effects are buffered by a cascade of chemical groups such as the NH3/NH4(+) group, leading to relatively stable pH values in both anolyte and catholyte. The transport of NH4(+) ions accounts for ~90% of the total current, thus quantitatively confirming that the NH4(+) ions serve as effective proton shuttles during MEC operations. Analysis further indicated that, because of the Donnan equilibrium at cation exchange membrane-anolyte/catholyte interfaces, the Na(+) ion in the anolyte actually facilitates the transport of NH4(+) ions during the early stage of a batch cycle and they compete with the NH4(+) ions weakly at later time. These insights, along with a new and simple method for predicting the strength of ammonia diffusion from the catholyte toward the anolyte, will help effective design and operation of bioeletrochemical system-based ammonia recovery systems.
Project description:Microbial electrochemical technologies have been extensively employed for phenol removal. Yet, previous research has yielded inconsistent results, leaving uncertainties regarding the feasibility of phenol degradation under strictly anaerobic conditions using anodes as sole terminal electron acceptors. In this study, we employed high-performance liquid chromatography and gas chromatography-mass spectrometry to investigate the anaerobic phenol degradation pathway. Our findings provide robust evidence for the purely anaerobic degradation of phenol, as we identified benzoic acid, 4-hydroxybenzoic acid, glutaric acid, and other metabolites of this pathway. Notably, no typical intermediates of the aerobic phenol degradation pathway were detected. One-chamber reactors (+0.4 V vs. SHE) exhibited a phenol removal rate of 3.5 ± 0.2 mg L-1 d-1, while two-chamber reactors showed 3.6 ± 0.1 and 2.6 ± 0.9 mg L-1 d-1 at anode potentials of +0.4 and + 0.2 V, respectively. Our results also suggest that the reactor configuration certainly influenced the microbial community, presumably leading to different ratios of phenol consumers and microorganisms feeding on degradation products.
Project description:Overexpression of the adenylate cyclase gene cyaC facilitates current generation by Shewanella oneidensis in bioelectrochemical systems