Interspecies Electron Transfer Mechanisms Research Articles

Residence Time Reduction in Anaerobic Reactors: Investigating the Economic Benefits of Magnetite-Induced Direct Interspecies Electron Transfer Mechanism

Existing research on direct interspecies electron transfer (DIET) has predominantly focused on the types and concentrations of conductive materials across diverse anaerobic digestion. However, insufficient understanding of the impact of residence time, a critical economic factor, prompted this investigation. Magnetite, a conductive material, was introduced into the anaerobic digestion of food wastewater, leading to a significant increase in ultimate methane production (Bu) with 25 mM-Fe3O4 (p < 0.05). Despite a subsequent decline in methane production efficiency from 388.9% to 7.1% over the 15- to 65-day anaerobic digestion period, the initial impact of increased methane production due to magnetite addition was evident. Control’s maximum methane production rate (Rm) was 27.5 mL/day, reaching its highest point at 37.4 mL/day with 15 mM-Fe3O4, accompanied by a noteworthy 56.6% reduction in the attainment day of Rm (Rm-day), shortened to 8.2 days. Even with 100 mM-Fe3O4, while Bu showed no significant difference, Rm-day exhibited a substantial reduction of 22.8. Despite the lower overall anaerobic digestion efficiency under some magnetite input conditions, this study confirmed a substantial shortening of Rm-day, suggesting that the DIET mechanism induced by conductive materials such as magnetite could reduce the residence time in continuous-type anaerobic reactors, contributing to improved economic feasibility.

Enhanced Anaerobic Digestion Using Conductive Materials through Mediation of Direct Microbial Interspecies Electron Transfer: A Review

The anaerobic digestion (AD) of organic matter is susceptible to the challenges posed by low-speed electron transfer between microorganisms and the limitation of low hydrogen partial pressure, resulting in low methane recovery efficiency and poor system stability. Numerous studies in recent years have shown that a variety of conductive materials can significantly increase the interspecies electron transfer (IET) rate, optimize the structure and function of anaerobic microbial communities, improve methane yield, and promote system stability by mediating the direct interspecies electron transfer (DIET) of reciprocal microorganisms. In this study, on the basis of investigating the IET mechanism of methanogenic microorganisms in the AD of organic matter, the effects of carbon-based conductive materials (activated carbon, biochar, carbon cloth, carbon fiber, graphite, graphite felt, graphene, and carbon nanotubes) and iron-based conductive materials (magnetite, Fe3O4, hematite, Fe2O3, goethite, and zero-valent iron) on AD performance and microbial community using DIET are reviewed. Future research should focus on establishing an evaluation system, identifying flora with DIET potential, and finding methods for engineering applications that increase recovery efficiency and reveal the principle of conductive materials to mediate DIET.

Meta-analysis of bio-based carbon materials for anaerobic digestion with direct interspecies electron transfer mechanism

There has been considerable interest in enhancing the anaerobic digestion process leading to the use of various additives or accelerants to leverage lucrative synergies, such as increased CH4 output and better digestate quality. There is less operational instability and substrate-induced inhibition with carbon-based materials, which helps the organic matter to break down quickly. New developments are examined in this research study to improve anaerobic digestion system performance, emphasizing cheap carbon materials used in anaerobic digestion, with special emphasis on using bio-based carbons like biochar, activated carbon and cloth made from bio-based coals. When these are used in an anaerobic digestion system, bio-based carbons help to promote bacterial syntropy along with speeding up direct interspecies electron transfer (DIET) and increasing methane production. This study emphasizes the production of bio-based carbon compounds and their use as anaerobic digestion additives with synergetic mechanisms. It also highlightsimprovement in biogas output using bio-based carbons for energy generation and sustainable environment leading to healthy environment.

Biofuel production from fruit and vegetable market waste and mature landfill leachate by an active filter-anaerobic digestion integrated system

• The sustainable management of landfill leachate (LL) is particularly problematic. • Anaerobic digestion (AD) is suitable for producing a sustainable biofuel. • Pre-treated LL is used for integration of NH 3 -N in AD of market waste. • The addition of granular activated carbon (GAC) was tested during AD. • GAC allowed a 14% increase of methane yield that reached 0.302 NL/gVS added . The management of municipal solid wastes represents one of the biggest challenges for the actual implementation of the circular economy. While the main “dry” fractions (such as plastics and paper) can be easily recycled, treatments for both the valorisation of the organic waste and for the disposal of leachate (produced by the landfilling of unsorted municipal waste) often turn out to be inadequate and expensive. Particularly, the leachate management is a critical issue since the mature landfill leachate (MLL) must be treated for decades even after the landfill closure. In this study, an innovative approach is proposed and it consists in an integrated system of active filters and anaerobic reactors for the simultaneous treatment of both waste streams (i.e., organic waste and MLL). Firstly, bio-refractory compounds (e.g., metals) present in MLL are removed by filtration on active materials (i.e., zero valent iron (ZVI) and granular activated carbon (GAC) or ZVI and Lapillus mixtures). The so-treated MLLs are then used as nutrient solutions for the anaerobic digestion (AD) of market wastes (MW) to optimize the C/N ratio. Moreover, during the anaerobic digestion, granular active carbon (GAC) is added in some reactors along with the feeding solution (substrate and pre-treated MLLs) to retain possible refractory or toxic compounds and to facilitate microorganisms’ activities by promoting the direct interspecies electron transfer mechanism. MLLs, used as nutrient solutions, lead to a stable AD of MW with a methane yield of about 0.260 NL/gVS added . GAC is proved to be also efficient in the enhancement of the methane production as higher methane yields (about 0.302 NL/gVS added ) are obtained in reactors where it is added as a supplement.

Proteomic Analysis of a Syntrophic Coculture of Syntrophobacter fumaroxidans MPOBT and Geobacter sulfurreducens PCAT.

We established a syntrophic coculture of Syntrophobacter fumaroxidans MPOBT (SF) and Geobacter sulfurreducens PCAT (GS) growing on propionate and Fe(III). Neither of the bacteria was capable of growth on propionate and Fe(III) in pure culture. Propionate degradation by SF provides acetate, hydrogen, and/or formate that can be used as electron donors by GS with Fe(III) citrate as electron acceptor. Proteomic analyses of the SF-GS coculture revealed propionate conversion via the methylmalonyl-CoA (MMC) pathway by SF. The possibility of interspecies electron transfer (IET) via direct (DIET) and/or hydrogen/formate transfer (HFIT) was investigated by comparing the differential abundance of associated proteins in SF-GS coculture against (i) SF coculture with Methanospirillum hungatei (SF-MH), which relies on HFIT, (ii) GS pure culture growing on acetate, formate, hydrogen as propionate products, and Fe(III). We noted some evidence for DIET in the SF-GS coculture, i.e., GS in the coculture showed significantly lower abundance of uptake hydrogenase (43-fold) and formate dehydrogenase (45-fold) and significantly higher abundance of proteins related to acetate metabolism (i.e., GltA; 62-fold) compared to GS pure culture. Moreover, SF in the SF-GS coculture showed significantly lower abundance of IET-related formate dehydrogenases, Fdh3 (51-fold) and Fdh5 (29-fold), and the rate of propionate conversion in SF-GS was 8-fold lower than in the SF-MH coculture. In contrast, compared to GS pure culture, we found lower abundance of pilus-associated cytochrome OmcS (2-fold) and piliA (5-fold) in the SF-GS coculture that is suggested to be necessary for DIET. Furthermore, neither visible aggregates formed in the SF-GS coculture, nor the pili-E of SF (suggested as e-pili) were detected. These findings suggest that the IET mechanism is complex in the SF-GS coculture and can be mediated by several mechanisms rather than one discrete pathway. Our study can be further useful in understanding syntrophic propionate degradation in bioelectrochemical and anaerobic digestion systems.

Enhanced anaerobic co-digestion of waste activated sludge and food waste by sulfidated microscale zerovalent iron: Insights in direct interspecies electron transfer mechanism

The enhancement of zerovalent iron (ZVI) on anaerobic digestion (AD) has been proved, but there are still some problems that constrain the large-scale application of ZVI, such as the destruction of cell membrane and the inhibition of methanogenesis led by rapid H2 accumulation. Aiming at these problems, sulfidated microscale zerovalent iron (S-mZVI) was employed to evaluate its effect on anaerobic co-digestion (AcoD) of waste activated sludge (WAS) and food waste (FW). Experimental results showed that S-mZVI promoted the direct interspecies electron transfer (DIET) between specific bacteria and methanogens, resulting in higher methane yield. At S-mZVI 10 g/L, the cumulative methane yield and ETS activity reached 264.78 mL/g-VS and 24.62 mg INTF/(g-TS h), which was 1.33 and 1.83 times that of blank. Microbiological analysis demonstrated that the abundance of DIET-related microorganisms such as Syntrophomonas, Methanosarcina and Methanobacterium increased with the increasing dosage of S-mZVI.

Improved Food Waste Stabilization and Valorization by Anaerobic Digestion Through Supplementation of Conductive Materials and Trace Elements

The positive effects of conductive material supplementation on anaerobic digestion have been mainly investigated for single synthetic substrates, while its significance for real and complex organic wastes such as food waste has not been sufficiently investigated. This study investigated the effect of conductive material (biochar and magnetite) and trace metal supplementation on the anaerobic digestion of food waste by means of biochemical methane potential assays. The results indicated that the supplementation of biochar and trace metals improved both total biogas production and methane yields. A biochar dose of 2.0 and 5.0 g/L resulted in 11.2 ± 6.5 and 27.3 ± 9.5% increase in biogas and 8.3 ± 6.8 and 33.2 ± 2.8% increase in methane yield, respectively. Moreover, the same reactors demonstrated high food waste stabilization performance of over 80% chemical oxygen demand removal efficiency. These results indicate that biochar supplementation leads to more enhanced anaerobic digestion operation that could be through increased surface area for microbial growth and/or direct interspecies electron transfer mechanism. In turn, food waste will not only be stabilized but also valorized by anaerobic digestion at higher efficiencies that support sustainable waste management through both environmentally safe disposal and value-added generation.

Dark fermentation production of volatile fatty acids from glucose with biochar amended biological consortium

Effects of adding biochars on dark fermentation production of volatile fatty acids (VFAs) from glucose were investigated. Nine biochars were synthesized and applied, together with an activated carbon, as the testing amendment to enhance preferable fermentation. Biochars were porous materials with internal pores and excess surface functional groups, which would lead to enrichment of acetate over butyrate in the VFA production. Biochar coconut and biochar longan shell showed excess functional groups and high bulk internal crystallinity, presented 109.6% and 71.8% enrichments of acetate production, respectively. The syntrophic growth of fermentative bacteria and homoacetogens on biochar surfaces via direct interspecies electron transfer mechanism was assumed to interpret the noted enhanced acetate production. The excess functional groups on biochar surface to facilitate biofilm development and the high crystallinity of biochar bulk to ease electron transfer favored the production of acetate.

Quantitative analysis of acetate oxidation in the presence of iron in a thermophilic methanogenic reactor

In natural and engineered anaerobic ecosystems, syntrophic oxidation of short-chain fatty acid is one of the key factors and interspecies electron transfer mechanisms between bacteria and archaea play a pivotal role. Growing evidence suggests that microorganisms have the capability to promote interspecies electrons transfer in a more direct manner. In the current study, three different types of iron oxides were used to thermophilic anaerobic reactors (at 50 °C), quantitative polymerase chain reaction and 16S rRNA sequencing technique were applied to investigate the relation between iron reduction bacteria and syntrophic acetate oxidation bacteria. Experiment results showed that goethite and hematite promoted the methane production and generation rate and partially relieved the negative effects of H2. The syntrophic relation between Syntrophaceticus and Methanobacterium worked with a direct interspecies electron transfer model rather than interspecies hydrogen transfer in goethite and hematite dosed reactors. The gene copies of Syntrophaceticus predominated over that of Geobacter. Theoretical calculations revealed that the rates of iron oxide mediated interspecies electron transfer among syntrophic partners were 105-106 times higher than those attainable via interspecies H2 transfer.

Elucidating Syntrophic Butyrate-Degrading Populations in Anaerobic Digesters Using Stable-Isotope-Informed Genome-Resolved Metagenomics.

Linking the genomic content of uncultivated microbes to their metabolic functions remains a critical challenge in microbial ecology. Resolving this challenge has implications for improving our management of key microbial interactions in biotechnologies such as anaerobic digestion, which relies on slow-growing syntrophic and methanogenic communities to produce renewable methane from organic waste. In this study, we combined DNA stable-isotope probing (SIP) with genome-centric metagenomics to recover the genomes of populations enriched in 13C after growing on [13C]butyrate. Differential abundance analysis of recovered genomic bins across the SIP metagenomes identified two metagenome-assembled genomes (MAGs) that were significantly enriched in heavy [13C]DNA. Phylogenomic analysis assigned one MAG to the genus Syntrophomonas and the other MAG to the genus Methanothrix. Metabolic reconstruction of the annotated genomes showed that the Syntrophomonas genome encoded all the enzymes for beta-oxidizing butyrate, as well as several mechanisms for interspecies electron transfer via electron transfer flavoproteins, hydrogenases, and formate dehydrogenases. The Syntrophomonas genome shared low average nucleotide identity (<95%) with any cultured representative species, indicating that it is a novel species that plays a significant role in syntrophic butyrate degradation within anaerobic digesters. The Methanothrix genome contained the complete pathway for acetoclastic methanogenesis, indicating that it was enriched in 13C from syntrophic acetate transfer. This study demonstrates the potential of stable-isotope-informed genome-resolved metagenomics to identify in situ interspecies metabolic cooperation within syntrophic consortia important to anaerobic waste treatment as well as global carbon cycling.IMPORTANCE Predicting the metabolic potential and ecophysiology of mixed microbial communities remains a major challenge, especially for slow-growing anaerobes that are difficult to isolate. Unraveling the in situ metabolic activities of uncultured species may enable a more descriptive framework to model substrate transformations by microbiomes, which has broad implications for advancing the fields of biotechnology, global biogeochemistry, and human health. Here, we investigated the in situ function of mixed microbiomes by combining stable-isotope probing with metagenomics to identify the genomes of active syntrophic populations converting butyrate, a C4 fatty acid, into methane within anaerobic digesters. This approach thus moves beyond the mere presence of metabolic genes to resolve "who is doing what" by obtaining confirmatory assimilation of the labeled substrate into the DNA signature. Our findings provide a framework to further link the genomic identities of uncultured microbes with their ecological function within microbiomes driving many important biotechnological and global processes.

Insights into direct interspecies electron transfer mechanisms for acceleration of anaerobic digestion of wastes

Anaerobic digestion of waste organic biomass is a well-known process for conversion of biomass to bioenergy. However, global acceptability of anaerobic digestion process for renewable energy has often been undermined because of its inconsistent or lower rate of biogas production. The overall success of the anaerobic digestion process considerably depends on the interactions between the microbial communities within the digester. The electron transfer through interspecies hydrogen transfer between acetogens and methanogens is a major bottleneck for successful anaerobic digestion process. Recent studies have reported the role of biotic (pili and cytochromes) as well as abiotic (conductive materials) components to accelerate direct interspecies electron transfer between microbial communities in the digester. These transfer mechanisms via biotic components of exoelectrogenic bacteria and methanogenic archaea is thermodynamically more favorable over indirect interspecies electron transfer. However, the use of conductive materials in promoting anaerobic digestion process has been an area of research in the last few years. The process of direct interspecies electron transfer promoting anaerobic digestion process has been investigated evidently in pure microbial cultures; however, there are scanty reports on existence of direct interspecies electron transfer in mixed microbial consortia. The present review highlights the fundamentals and applications of direct interspecies electron transfer-promoted anaerobic digestion process at laboratory scale studies used to improve kinetics of methanogenesis.

Magnetite particles triggering a faster and more robust syntrophic pathway of methanogenic propionate degradation.

Interspecies electron transfer mechanisms between Bacteria and Archaea play a pivotal role during methanogenic degradation of organic matter in natural and engineered anaerobic ecosystems. Growing evidence suggests that in syntrophic communities electron transfer does not rely exclusively on the exchange of diffusible molecules and energy carriers such as hydrogen or formate, rather microorganisms have the capability to exchange metabolic electrons in a more direct manner. Here, we show that supplementation of micrometer-size magnetite (Fe3O4) particles to a methanogenic sludge enhanced (up to 33%) the methane production rate from propionate, a key intermediate in the anaerobic digestion of organic matter and a model substrate to study energy-limited syntrophic communities. The stimulatory effect most probably resulted from the establishment of a direct interspecies electron transfer (DIET), based on magnetite particles serving as electron conduits between propionate-oxidizing acetogens and carbon dioxide-reducing methanogens. Theoretical calculations revealed that DIET allows electrons to be transferred among syntrophic partners at rates which are substantially higher than those attainable via interspecies H2 transfer. Besides the remarkable potential for improving anaerobic digestion, which is a proven biological strategy for renewable energy production, the herein described conduction-based DIET could also have a role in natural methane emissions from magnetite-rich soils and sediments.

Characterization and modelling of interspecies electron transfer mechanisms and microbial community dynamics of a syntrophic association

Syntrophic associations are central to microbial communities and thus have a fundamental role in the global carbon cycle. Despite biochemical approaches describing the physiological activity of these communities, there has been a lack of a mechanistic understanding of the relationship between complex nutritional and energetic dependencies and their functioning. Here we apply a multi-omic modelling workflow that combines genomic, transcriptomic and physiological data with genome-scale models to investigate dynamics and electron flow mechanisms in the syntrophic association of Geobacter metallireducens and Geobacter sulfurreducens. Genome-scale modelling of direct interspecies electron transfer reveals insights into the energetics of electron transfer mechanisms. While G. sulfurreducens adapts to rapid syntrophic growth by changes at the genomic and transcriptomic level, G. metallireducens responds only at the transcriptomic level. This multi-omic approach enhances our understanding of adaptive responses and factors that shape the evolution of syntrophic communities.

Microbial interspecies electron transfer via electric currents through conductive minerals

In anaerobic biota, reducing equivalents (electrons) are transferred between different species of microbes [interspecies electron transfer (IET)], establishing the basis of cooperative behaviors and community functions. IET mechanisms described so far are based on diffusion of redox chemical species and/or direct contact in cell aggregates. Here, we show another possibility that IET also occurs via electric currents through natural conductive minerals. Our investigation revealed that electrically conductive magnetite nanoparticles facilitated IET from Geobacter sulfurreducens to Thiobacillus denitrificans, accomplishing acetate oxidation coupled to nitrate reduction. This two-species cooperative catabolism also occurred, albeit one order of magnitude slower, in the presence of Fe ions that worked as diffusive redox species. Semiconductive and insulating iron-oxide nanoparticles did not accelerate the cooperative catabolism. Our results suggest that microbes use conductive mineral particles as conduits of electrons, resulting in efficient IET and cooperative catabolism. Furthermore, such natural mineral conduits are considered to provide ecological advantages for users, because their investments in IET can be reduced. Given that conductive minerals are ubiquitously and abundantly present in nature, electric interactions between microbes and conductive minerals may contribute greatly to the coupling of biogeochemical reactions.