Microbial Electrosynthesis Research Articles

Methanol as a co-substrate with CO2 enhances butyrate production in microbial electrosynthesis

Methanol is a promising feedstock for the bio-based economy as it can be derived from organic waste streams or produced electrochemically from CO2. Acetate production from CO2 in microbial electrosynthesis (MES) has been widely studied, while more valuable compounds such as butyrate are currently attracting attention. In this study, methanol was used as a co-substrate with CO2 to enhance butyrate production in MES. Feeding with CO2 and methanol resulted in the highest butyrate production rates and titres of 0.36 ± 0.01 g L−1 d−1 and 8.6 ± 0.2 g L−1, respectively, outperforming reactors with only CO2 feeding (0.20 ± 0.03 g L−1 d−1 and 5.2 ± 0.1 g L−1, respectively). Methanol acted as electron donor and as carbon source, both of which contributed ca. 50% of the carbon in the products. Eubacterium was the dominant genus with 52.6 ± 2.5% relative abundance. Thus, we demonstrate attractive route for the use of the C1 substrates, CO2 and methanol, to produce mainly butyrate.Key points• Butyrate was the main product from methanol and CO2in MES• Methanol acted as both carbon and electron source in MES• Eubacterium dominating microbial culture was enriched in MESGraphical

Bioelectrochemical protein production valorising NH3-rich pig manure-derived wastewater and CO2 from anaerobic digestion

Microbial electrosynthesis (MES) cell use is an innovative approach for single-cell proteins (SCP) production. Coupling MES with the valorization of CO2 from anaerobic digestion and nitrogen from livestock effluents has beneficial environmental effects, reducing greenhouse gas emissions and nitrogen overloading. In addition, the reducing power needed can come from surplus renewable energy. In this study, MES with a biochar-functionalized cathode was tested at varying polarizations, i.e. non polarized, −0.6 V and −1.0 V vs Ag/AgCl, and biogas-derived CO2 and recovered ammonia from pig slurry was supplied.Negative polarization switched the microbial community from heterotrophic, typical of unpolarized MES, to a mix of both heterotrophic and autotrophic/electrotrophic communities at −0.6 V and to mainly autotrophic/electrotrophic at −1.0 V. The more negative polarization allowed the highest CO2 and N capture, i.e. 39 ± 2 % of the supplied CO2, and 6.7 ± 0.8 % supplied N. Microbial biomass characterization indicated a protein content on dry matter basis of 33.1 ± 1.3 % (unpolarized), 43.2 ± 0.6 % (−0.6 V) and 69.1 ± 1.0 % (−1.0 V). The amino acids profiles investigated showed a high nutritional value of the produced biomass, not far from those of conventional protein sources used for producing feed/food.

In-situ biogenic FeS boosted acetate accumulation through CO2 capture and valorization using microbial electrosynthesis (MES)

Biosynthetic strategy of ferrous sulfide (FeS) through Shewanella oneidensis MR-1 in bioelectrochemical systems has been extensively explored to maintain bettered electron transfer performance. In this study, the simultaneous FeS biosynthesis and CO2 reduction were conducted in the microbial electrosynthesis (MES) system, aiming at improving electron transfer efficiency as well as acetate production. It was shown that the FeS was successfully biomineralized and then loaded onto the sludge in the Fe3+/S2O32−/MR-1 group, of which the acetate accumulation increased by 87.50%, 64.63% and 33.66% than the control, MR-1 and Fe3+/S2O32− groups. Also, electrochemical performance revealed by the CV analysis was enhanced, whereas the increased EPS accumulation and intracellular electron transfer activity by the biosynthesized FeS could also be proved. Moreover, the biogenic FeS enriched the abundances of acetogenic and sulfur-utilizing species as Acetobacterium, Clostridium_sensu_stricto_13, Sulfuricurvum and Desulfovibrio. Meanwhile, strongly positive correlations between Shewanella, the sulfur utilizers and acetogens were confirmed in the Fe3+/S2O32−/MR-1 group. With improved electron transfer efficiency and the boosted acetate accumulation, the in-situ biogenic FeS might be promising for CO2 capture and utilization.

Comparing the performance of fluidized and fixed granular activated carbon beds as cathodes for microbial electrosynthesis of carboxylates from CO2

Microbial electrosynthesis (MES) can use renewable electricity to power microbial conversion of carbon dioxide (CO2) into carboxylates. To ensure high productivities in MES, good mass transfer must be ensured, which could be accomplished with fluidization of granular activated carbon (GAC). In this study, fluidized and fixed GAC bed cathodes were compared. Acetate production rate and current density were 42 % and 47 % lower, respectively, in fluidized than fixed bed reactors. Although similar microbial consortium dominated by Eubacterium and Proteiniphilum was observed, lowest biomass quantity was measured with fixed GAC bed indicating higher specific acetate production rates compared to fluidized GAC bed. Furthermore, charge efficiency was the highest and charge recovery in carboxylates the lowest in fixed GAC beds indicating enhanced hydrogen evolution and need for enhancing CO2 feeding to enable higher production rates of acetate. Overall, fixed GAC beds have higher efficiency for acetate production in MES than fluidized GAC beds.

Microbial electrosynthesis of valuable chemicals from the reduction of CO2: a review.

The present energy demand of the world is increasing but the fossil fuels are gradually depleting. As a result, the need for alternative fuels and energy sources is growing. Fuel cells could be one alternative to address the challenge. The fuel cell can convert CO2 to value-added chemicals. The potential of bio-fuel cells, specifically enzymatic fuel cells and microbial fuel cells, and the importance of immobilization technology in bio-fuel cells are highlighted. The review paper also includes a detailed explanation of the microbial electrosynthesis system to reduce CO2 and the value-added products during microbial electrosynthesis. Future research in bio-electrochemical synthesis for CO2 conversion is expected to prioritize enhancing biocatalyst efficiency, refining reactor design, exploring novel electrode materials, understanding microbial interactions, integrating renewable energy sources, and investigating electrochemical processes for carbon capture and selective CO2 reduction. The challenges and perspectives of bio-electrochemical systems in the application of CO2 conversion are also discussed. Overall, this review paper provides valuable insights into the latest developments and criteria for effective research and implementation in bio-fuel cells, immobilization technology, and microbial electro-synthesis systems.

Using a non-precious metal catalyst for long-term enhancement of methane production in a zero-gap microbial electrosynthesis cell

Microbial electrosynthesis (MES) cells exploit the ability of microbes to convert CO2 into valuable chemical products such as methane and acetate, but high rates of chemical production may need to be mediated by hydrogen and thus require a catalyst for the hydrogen evolution reaction (HER). To avoid the usage of precious metal catalysts and examine the impact of the catalyst on the rate of methane generation by microbes on the electrode, we used a carbon felt cathode coated with NiMo/C and compared performance to a bare carbon felt or a Pt/C-deposited cathode. A zero-gap configuration containing a cation exchange membrane was developed to produce a low internal resistance, limit pH changes, and enhance direct transport of H2 to microorganisms on the biocathode. At a fixed cathode potential of –1 V vs Ag/AgCl, the NiMo/C biocathode enabled a current density of 23 ± 4 A/m2 and a high methane production rate of 4.7 ± 1.0 L/L-d. This performance was comparable to that using a precious metal catalyst (Pt/C, 23 ± 6 A/m2, 5.4 ± 2.8 L/L-d), and 3–5 times higher than plain carbon cathodes (8 ± 3 A/m2, 1.0 ± 0.4 L/L-d). The NiMo/C biocathode was operated for over 120 days without observable decay or severe cathode catalyst leaching, reaching an average columbic efficiency of 53 ± 9 % based on methane production under steady state conditions. Analysis of microbial community on the biocathode revealed the dominance of the hydrogenotrophic genus Methanobacterium (∼40 %), with no significant difference found for biocathodes with different materials. These results demonstrated that HER catalysts improved rates of methane generation through facilitating hydrogen gas evolution to an attached biofilm, and that the long-term enhancement of methane production in MES was feasible using a non-precious metal catalyst and a zero-gap cell design.

Electrodes from carbonized grass clippings for bioelectrochemical systems

One major obstacle to the commercialization of electrobiotechnological systems is the cost of materials, including expensive electrodes. Smart recycling as well as the use of renewable resources can contribute to producing electrodes more ecologically and economically. Green waste is a biogenic residual material that occurs mainly in urban areas and is currently not recycled to a sufficient extent. Here we show the fabrication of electrodes from carbonized grass clippings and their application in microbial electrosynthesis as well as microbial fuel cells. While the electrodes cannot compete with established metal competitors for water electrolysis in microbial electrosynthesis, they perform comparably to commercial graphite electrodes in microbial fuel cells. With Geobacter sulfurreducens, a current response can be recorded for more than six weeks. To the best of our knowledge, this is the first time that carbonized green waste has been used as an electrode material for bioelectrochemical systems. This demonstrates the potential of carbonized biological materials as a raw material for electrodes and presents a recycling alternative for green waste.

Integration of biocompatible hydrogen evolution catalyst developed from metal-mix solutions with microbial electrosynthesis

Microbial conversion of CO2 to multi-carbon compounds such as acetate and butyrate is a promising valorisation technique. For those reactions, the electrochemical supply of hydrogen to the biocatalyst is a viable approach. Earlier we have shown that trace metals from microbial growth media spontaneously form in situ electro-catalysts for hydrogen evolution. Here, we show biocompatibility with the successful integration of such metal mix-based HER catalyst for immediate start-up of microbial acetogenesis (CO2 to acetate). Also, n-butyrate formation started fast (after twenty days). Hydrogen was always produced in excess, although productivity decreased over the 36 to 50 days, possibly due to metal leaching from the cathode. The HER catalyst boosted microbial productivity in a two-step microbial community bioprocess: acetogenesis by a BRH-c20a strain and acetate elongation to n-butyrate by Clostridium sensu stricto 12 (related) species. These findings provide new routes to integrate electro-catalysts and micro-organisms showing respectively bio and electrochemical compatibility.

A biocompatible electrode/exoelectrogens interface augments bidirectional electron transfer and bioelectrochemical reactions

Bidirectional electron transfer is about that exoelectrogens produce bioelectricity via extracellular electron transfer at anode and drive cytoplasmic biochemical reactions via extracellular electron uptake at cathode. The key factor to determine above bioelectrochemical performances is the electron transfer efficiency under biocompatible abiotic/biotic interface. Here, a graphene/polyaniline (GO/PANI) nanocomposite electrode specially interfacing exoelectrogens (Shewanella loihica) and augmenting bidirectional electron transfer was conducted by in-situ electrochemical modification on carbon paper (CP). Impressively, the GO/PANI@CP electrode tremendously improved the performance of exoelectrogens at anode for wastewater treatment and bioelectricity generation (about 54 folds increase of power density compared to blank CP electrode). The bacteria on electrode surface not only showed fast electron release but also exhibited high electricity density of extracellular electron uptake through the proposed direct electron transfer pathway. Thus, the cathode applications of microbial electrosynthesis and bio-denitrification were developed via GO/PANI@CP electrode, which assisted the close contact between microbial outer-membrane cytochromes and nanocomposite electrode for efficient nitrate removal (0.333 mM/h). Overall, nanocomposite modified electrode with biocompatible interfaces has great potential to enhance bioelectrochemical reactions with exoelectrogens.

Production of polyhydroxybutyrate from industrial flue gas by microbial electrosynthesis

The aim of this work was to produce the biopolymer polyhydroxybutyrate (PHB) from industrial flue gas as CO2 source and electrolysis originating hydrogen via microbial electrosynthesis. Besides the laboratory experiments, the experiments were carried out under industrial conditions directly on-site in a cogeneration plant. We were able to demonstrate that the use of flue gas as a CO2 source has no detectable negative effect on bacterial growth and PHB production in comparison to a pure gas mixture. In an electrochemical H-cell 333 ± 44 mg L−1 PHB were obtained, which corresponds to a PHB content of 43 ± 3 % of the cell dry weight. By using flue gas for the production of the biopolymer PHB, not only CO2 emissions are reduced, but also possible environmental pollution from non-biodegradable plastics. To the best of our knowledge, this is the first time that the production of PHB from flue gas has been demonstrated using Cupriavidus necator.

Chain elongation in continuous microbial electrosynthesis cells: The effect of pH and precursors supply

Microbial electrosynthesis (MES) enables the production of carbon-neutral chemicals using CO2 as a carbon source. Acetic acid is the main MES product, but recent studies show the direct production of elongated carboxylic acids, e.g., butyric and caproic acid. However, the production of elongated acids in MES systems is still inefficient due to the low growth rates of acetogenic bacteria and to limited solventogenic rates. Subsequently, researchers have produced elongated carboxylic acids directly from acetic acid or have operated MES systems at low pH to favor solventogenesis. However, the effect the addition of different chain elongation precursors and the operation pH exerts in the bioelectrochemical production of elongated acids remains unclear. To investigate this, three pH-controlled MES systems were operated in this study with continuous liquid and gas supply. MES systems elongating acetic acid at pH 6 achieved higher butyric (0.71 vs. 0.42 g L−1) and caproic acid (0.71 vs. 0.42 g L−1) titers in the absence of CO2 sparging. Additionally, lowering the pH to 5 in the MES systems fed with CO2 and acetic acid improved the elongated acids titers, reaching 0.72 g L−1 butyric and 0.33 g L−1 caproic acid. The 16 S rRNA analysis showed the community was dominated by Oscillibacter at pH 6, and by Clostridium at pH 5. Furthermore, the first scanning electron microscopy pictures revealing biofilm stratification in MES cathodes were taken in this study, where homogeneous rod-shaped bacteria biofilm layers, in contact with the graphite cathode, were covered by heterogeneous biofilm layers.

Delineating cathodic extracellular electron transfer pathways in microbial electrosynthesis: Modulation of polarized potential and Pt@C addition

Microbial electrosynthesis (MES) is an innovative technology that employs microbes to synthesize chemicals by reducing CO2. A comprehensive understanding of cathodic extracellular electron transfer (CEET) is essential for the advancement of this technology. This study explores the impact of different cathodic potentials on CEET and its response to introduction of hydrogen evolution materials (Pt@C). Without the addition of Pt@C, H2-mediated CEET contributed up to 94.4 % at −1.05 V. With the addition of Pt@C, H2-mediated CEET contributions were 76.6 % (−1.05 V) and 19.9 % (−0.85 V), respectively. BRH-c20a was enriched as the dominated microbe (>80 %), and its relative abundance was largely affected by the addition of Pt@C NPs. This study highlights the tunability of MES performance through cathodic potential control and the addition of metal nanoparticles.

Shewanella oneidensis MR-1 coupled with biomass-derived carbon dots for promoted bioelectrochemical CO2 reduction: Mechanism elucidation of intensified energy metabolism

Bioelectrochemical CO2 reduction reaction (CO2RR) has been touted as one of the promising strategies to convert CO2 to high-value chemicals via the efficient microbial electrosynthesis (MES). However, the sluggish electron transfer efficiency between microbes and cathodes hampers the CO2 reduction efficiency. Herein, biomass-derived carbon dots (CDs) with high electrical conductivity were coupled with Shewanella oneidensis (S. oneidensis) MR-1 to construct a nanomaterial-bacterial biohybrid system to promote CO2-to-formate formation. Specifically, the corncob-derived CDs (cc-CDs) were highly-biocompatible with S. oneidensis MR-1, with such a constructed biohybrid system showing a marked formate titer of 0.50 mmol/L. Upon doping with Fe element, the resultant Fe-cc-CDs exhibited a considerably boosted formate titer of 1.22 mmol/L, which was 7.7 times higher than the pure S. oneidensis MR-1 system. Such significant CO2-to-formate conversion enhancement was contributed by the internalized CDs in S. oneidensis MR-1 stimulating extracellular electron transfer efficiency, therefore accelerating energy metabolism for formate bioelectrosynthesis. The intensified energy metabolism in S. oneidensis MR-1 was corroborated by the increased concentrations of ATP, NADH and overexpression of energy-related genes. This work demonstrates an intensified energy metabolism exemplification for CO2 reduction, which can be expanded to a host of different nanomaterials-internalized MES systems for bioelectrochemical implications.

ZnMo-MOF as anti-CO hydrogen electrocatalyst enhance microbial electrosynthesis for CO/CO2 conversion

Microbial electrosynthesis (MES) is an electrically driven technology that can be used for converting CO/CO2 into chemicals. The unique electronic and substrate properties of CO make it an important research target for MES. However, CO can poison the cathode and increase the overpotential of hydrogen evolution reaction (HER), thus reducing the electron transfer rate via H2. This work evaluated the effect of an anti-CO HER catalyst on the performance of MES for CO/CO2 conversion. ZnMo-metal–organic framework (MOF) materials with different calcination temperatures were synthesized. ZnMo-MOF-800 with Mo2C nanoparticles as active centers exhibited excellent resistance to CO toxicity. It also obtained the highest hydrogen evolution and enhanced electron transfer rate in CO atmosphere. MES with ZnMo-MOF-800 cathode and Clostridium ljungdahlii as biocatalyst obtained 0.31 g L−1 d−1 acetate yield, 0.1 g L−1 d−1 butyrate yield, and 0.09 g L−1 d−1 2,3-butanediol yield in CO/CO2, while Pt/C only get 0.076 g L−1 d−1 acetate yield, 0.05 g L−1 d−1 butyrate yield and 0.02 g L−1 d−1 2,3-butanediol yield. ZnMo-MOF-800 was conducive to biofilm formation, enabling it to better resist CO toxicity. This work provides new opportunities for constructing a highly efficient cathode with an anti-CO hydrogen evolution catalyst to enhance CO/CO2 conversion in MES.

Impact of intermittent electricity supply on a conceptual process design for microbial conversion of CO2 into hexanoic acid

Combining intermittent renewable electricity (IRE) with carbon capture and utilisation is urgently needed in the chemical sector. In this context, microbial electrosynthesis (MES) has gained attention. It can electrochemically produce hexanoic acid, a value-added chemical, from CO2. However, there is a lack of understanding regarding how the intermittency of renewable electricity could impact the design of a MES plant. We studied this using Aspen Plus models.A MES plant that was powered by constant grid electricity could operate from 100% down to 70% of its nominal capacity, at which point the heat exchangers and the internal geometrical design of the distillation towers became bottlenecks. The levelised production cost of hexanoic acid (LPCC6A) was estimated at 4.0 €/kg. Switching to IRE supply increased LPCC6A to 5.3 €/kg (for wind electricity) and 4.7 €/kg (for hybrid renewable electricity).A battery energy storage system (BESS) was deployed. The lowest LPCC6A was found at a BESS installation of 29 GJ/h for wind electricity (5.1 €/kg) and at 12 GJ/h for hybrid renewable electricity (4.7 €/kg). In both situations, the volume flexibility of the MES plant was not improved. At the investigated market and operating conditions, coupling IRE to the MES plant was economically infeasible.

Selective colonization of multifunctional microbes that facilitates caproate production in microbial electrosynthesis system

As promising and value-added platform chemicals, the biosynthesis of medium-chain fatty acids (MCFAs, C6-C12) via chain elongation (CE) in microbial electrosynthesis systems (MES) has recently garnered significant attentions due to the unique bioelectrocatalytic behaviors and the reduction of exogenous electron donors (EDs). Caproate-synthesizing bacteria (CSB) played a crucial role in the generation of acetyl-CoA, which subsequently underwent reverse β oxidation to synthesize MCFAs. However, the presence of electrochemically active bacteria (EAB) was also pivotal for MES as they generated hydrogen in the cathodic electron transfer pathways, the specific colonization patterns of key symbionts remained unknown. The aim of this study was to investigate the selective colonization of EAB and CSB on the performance of caproate production in MES. The reactors colonized EAB and CSB achieved the maximum caproate concentration of 5332.5 mg COD/L, which was 7.9 and 1.3 times higher compared to that inoculated with either sole EAB or CSB, respectively. Moreover, the reactors colonized with mixed culture exhibited a clear redox peak and preferable electron transfer efficiency. CSB and EAB mutually interacted in metabolic activities, thereby resulting in increased substrate utilization and caproate production. The findings of this study provide a new strategy and insights for value-added chemicals recovery in microbial electrosynthesis platform.

Enhancing Predictions of Acetate and Ethanol Production from Microbial Electrosynthesis Using Optimized Machine Learning Models

Enhancing Predictions of Acetate and Ethanol Production from Microbial Electrosynthesis Using Optimized Machine Learning Models

Nano zero-valent iron functioned 3D printing graphene aerogel electrode for efficient solar-driven biocatalytic methane production

To address the urgent need for developing carbon-neutral technologies, microbial electrosynthesis (MES), as an important contributor to the renewable energy field, can convert carbon dioxide to multi-carbon chemicals using microbes serving as living catalysts at the cathode. However, the performance of the cathode is restricted by the weak electrochemical reaction kinetics of the organisms, the specific area of the electrode, and the mass transfer under high current. Herein, we report a facile 3D-printed graphene aerogel (GA) electrode built with a nanozero-valent iron (NZVI)-functionalized graphene ink. The 3D-printed GA electrode was designed to possess hierarchical pores with an enhanced high specific surface area for microbial colonization while ensuring mass transfer. NZVI can assist in-situ hydrogen generation, facilitating biofilm formation and methane (CH4) production in the indirect electron transfer pathway. Consequently, the optimized 3D-printed electrode achieved a maximum CH4 production rate of 6993 ± 832 mmol/m2/d with a faradaic efficiency of 83.7%, which increased by 3.24-fold to the carbon felt electrode. In addition, integrating the proposed cathode into a PV-electrolyzer cell yielded a solar-to-CH4 efficiency of 4.70%. This study provides a unique method for the development of advanced cathodes and underscores the potential of integrating MES with renewable energy technologies.

Establishing CRISPRi for Programmable Gene Repression and Genome Evolution in Cupriavidus necator.

Cupriavidus necator H16 is a "Knallgas" bacterium with the ability to utilize various carbon sources and has been employed as a versatile microbial cell factory to produce a wide range of value-added compounds. However, limited genome engineering, especially gene regulation methods, has constrained its full potential as a microbial production platform. The advent of CRISPR/Cas9 technology has shown promise in addressing this limitation. Here, we developed an optimized CRISPR interference (CRISPRi) system for gene repression in C. necator by expressing a codon-optimized deactivated Cas9 (dCas9) and appropriate single guide RNAs (sgRNAs). CRISPRi was proven to be a programmable and controllable tool and could successfully repress both exogenous and endogenous genes. As a case study, we decreased the accumulation of polyhydroxyalkanoate (PHB) via CRISPRi and rewired the carbon fluxes to the synthesis of lycopene. Additionally, by disturbing the expression of DNA mismatch repair gene mutS with CRISPRi, we established CRISPRi-Mutator for genome evolution, rapidly generating mutant strains with enhanced hydrogen peroxide tolerance and robustness in microbial electrosynthesis (MES) system. Our work provides an efficient CRISPRi toolkit for advanced genetic manipulation and optimization of C. necator cell factories for diverse biotechnology applications.

Co3O4/C derived from ZIF-67 cathode enhances the microbial electrosynthesis of acetate from CO2

Microbial Electrosynthesis (MES) is an electrochemical reduction technology that efficiently converts CO2 into chemicals by electrically driving microbial catalysts. However, the lower mass transfer efficiency of CO2 and hydrogen H2 affected the performance of the MES. In this study, Co3O4/C is derived from the carbonization of ZIF-67 with different temperature (700 °C,800 °C,900 °C), in which some Co ions are exposed and a large number of catalytically active sites are supplied. Among them, the Co3O4/C with 800 °C had the highest hydrogen evolution reaction activity and the lowest internal resistance. The Co3O4/C with 800 °C showed the highest hydrogen production rate of 81.34 × 102 mol day−1, which was 4.4 times higher than that of the bare carbon felt. Furthermore, the Co3O4/C with 800 °C modified cathode was beneficial to the formation of biofilm and improved the enrichment of Acetobacterium and Arcobacter. The highest acetate yield of 0.19 g L−1d−1 was obtained by Co3O4/C with 800 °C, which was 2.1 times higher than that of bare carbon felt, and the corresponding acetate concentration reached 5.61 ± 0.1 g L−1. Co3O4/C derived from ZIF-67 with good H2 production and CO2 adsorption ability is an effective strategy to improve MES cathode.