Different outer membrane c‐type cytochromes are involved in direct interspecies electron transfer to Geobacter or Methanosarcina species

Direct interspecies electron transfer(DIET)may be most important in methanogenic environments,but mechanistic studies of DIET to date have primarily focused on cocultures in which fumarate was the terminal electron acceptor.To better understand DIET with methanogens,the transcriptome of Geobacter metallireducens during DIET‐based growth with G.sulfurreducens reducing fumarate was compared with G.metallireducens grown in coculture with diverse Methanosarcina.The transcriptome of G.metallireducens cocultured with G.sulfurreducens was significantly different from those with Methanosarcina.Furthermore,the transcriptome of G.metallireducens grown with Methanosarcina barkeri,which lacks outer‐surface c‐type cytochromes,differed from those of G.metallireducens cocultured with M.acetivorans or M.subterranea,which have an outer‐surface c‐type cytochrome that serves as an electrical connect for DIET.Differences in G.metallireducens expression patterns for genes involved in extracellular electron transfer were particularly notable.Cocultures with c‐type cytochrome deletion mutant strains,ΔGmet_0930,ΔGmet_0557 andΔGmet_2896,never became established with G.sulfurreducens but adapted to grow with all three Methanosarcina.Two porin–cytochrome complexes,PccF and PccG,were important for DIET;however,PccG was more important for growth with Methanosarcina.Unlike cocultures with G.sulfurreducens and M.acetivorans,electrically conductive pili were not needed for growth with M.barkeri.Shewanella oneidensis,another electroactive microbe with abundant outer‐surface c‐type cytochromes,did not grow via DIET.The results demonstrate that the presence of outer‐surface c‐type cytochromes does not necessarily confer the capacity for DIET and emphasize the impact of the electron‐accepting partner on the physiology of the electron‐donating DIET partner.

Combined pretreatment using CaO and liquid fraction of digestate of rice straw: Anaerobic digestion performance and electron transfer

To improve anaerobic digestion(AD)efficiency of rice straw,solid alkaline CaO and the liquid fraction of digestate(LFD)were used as pretreatment agents of rice straw.The results showed that AD performance of rice straw with CaOLFD pretreatment was optimal in different pretreatment methods of the CaO+LFD,CaOLFD,LFD+CaO,CaO,and LFD.The maximum methane yield(314 ml(g VS)^(-1))and the highest VFAs concentration(14851 mg·L^(-1) on day 3)of the CaOLFD pretreatment group were 81%and 118%higher than that of the control group,respectively.Under the action of solid alkaline CaO,the bacteria of Clostridium,Atopostipes,Sphaerochaeta,Tissierella,Thiopseudomonas,Rikenellaceae,and Sedimentibacter could build up co-cultures with the archaeal of Methanosaeta,Methanobacterium,and Methanosarcina performing direct interspecies electron transfer(DIET)and improving AD performance of rice straw.Therefore,the combined pretreatment using CaO and LFD could not only pretreat rice straw but also stimulate co-cultures of microorganism to establish DIET enhancing AD efficiency.

Comparative transcriptomic insights into the mechanisms of electron transfer in Geobacter co-cultures with activated carbon and magnetite

Both activated carbon and magnetite have been reported to promote the syntrophic growth of Geobacter metallireducens and Geobacter sulfurreducens co-cultures, the first model to show direct interspecies electron transfer (DIET); however, differential transcriptomics of the promotion on co-cultures with these two conductive materials are unknown. Here, the comparative transcriptomic analysis of G. metallireducens and G. sulfurreducens co-cultures with granular activated carbon (GAC) and magnetite was reported. More than 2.6-fold reduced transcript abundances were determined for the uptake hydrogenase genes of G. sulfurreducens as well as other hydrogenases in those co-cultures to which conductive materials had been added. This is consistent with electron transfer in G. metallireducens-G. sulfurreducens co-cultures as evinced by direct interspecies electron transfer (DIET). Transcript abundance for the structural component of electrically conductive pili (e-pili), PilA, was 2.2-fold higher in G. metallireducens, and, in contrast, was 14.9-fold lower in G. sulfurreducens in co-cultures with GAC than in Geobacters co-cultures without GAC. However, it was 9.3-fold higher in G. sulfurreducens in co-cultures with magnetite than in Geobacters co-cultures. Mutation results showed that GAC can be substituted for the e-pili of both strains but magnetite can only compensate for that of G. sulfurreducens, indicating that the e-pili is a more important electron acceptor for the electron donor strain of G. metallireducens than for G. sulfurreducens. Transcript abundance for G. metallireducens c-type cytochrome gene GMET_RS14535, a homologue to c-type cytochrome gene omcE of G. sulfurreducens was 9.8-fold lower in co-cultures with GAC addition, while that for OmcS of G. sulfurreducens was 25.1-fold higher in co-cultures with magnetite, than in that without magnetite. Gene deletion studies showed that neither GAC nor magnetite can completely substitute the cytochrome (OmcE homologous) of G. metallireducens but compensate for the cytochrome (OmcS) of G. sulfurreducens. Moreover, some genes associated with central metabolism were up-regulated in the presence of both GAC and magnetite; however, tricarboxylic acid cycle gene transcripts in G. sulfurreducens were not highly-expressed in each of these amended co-cultures, suggesting that there was considerable redundancy in the pathways utilised by G. sulfurreducens for electron transfer to reduce fumarate with the amendment of GAC or magnetite. These results support the DIET model of G. metallireducens and G. sulfurreducens and suggest that e-pili and cytochromes of the electron donor strain are more important than that of the electron acceptor strain, indicating that comparative transcriptomics may be a promising route by which to reveal different responses of electron donor and acceptor during DIET in co-cultures.

Biogeochemistry of methanogenesis with a specific emphasis on the mineral-facilitating effects

The Earth surface contains various oxic and anoxic environments. The later include natural wetlands,river and lake sediments, paddy field soils and landfills. In the last few decades, the biogeochemical cycle of carbon in anoxic environments, which leads to the production and emission of methane, a potent greenhouse gas in the atmosphere, has drawn great attentions from both scientific and public sectors. New organisms and mechanisms involved in methanogenesis and carbon cycling have been uncovered. Interspecies electron transfer is considered as a crucial step in methanogenesis in anoxic environments.Electron-carrying mediators, like H_2 and formate, are known to play the key role in electron transfer. Recently, it has been found that in addition to the conventional electron transfer via chemical mediators, direct interspecies electron transfer(DIET) can occur. In this Review, we describe the ecology and biogeochemistry of methanogenesis and highlight the effect of microbe-mineral interaction on microbial syntrophy. Recent advances in the study of DIET may pave the way towards a mechanistic understanding of methanogenesis and the influence of microbe-mineral interaction on this process.

The role of NiFe_(2)O_(4)nanoparticle in the anaerobic digestion(AD)of waste activated sludge(WAS)

Anaerobic digestion(AD)is a promising technology for the treatment of waste activated sludge(WAS)with energy recovery.However,the low methane yield and slow methanogenesis limit its broad application.In this study,the NiFe_(2)O_(4)nanoparticles(NPs)were fabricated and applied as a conductive material to enhance the AD via promoting the direct interspecies electron transfer(DIET).The crystal structure,specific surface area,morphology and elemental composition of the as-prepared NiFe_(2)O_(4)NPs were characterized by X-ray diffraction(XRD),Brunauer-Emmett-Teller(BET),scanning electron microscopy(SEM)and energy dispersive spectroscopy(EDS).The biochemical methane potential(BMP)test was performed(lasting for 35 days)to evaluate the energy recovery in AD with the addition of the NiFe_(2)O_(4)NPs.The results illustrate that NiFe_(2)O_(4)NPs could accelerate both the hydrolysis,acidogenesis and methanogenesis,i.e.,the cumulative methane production and daily methane yield increased from 96.76±1.70 mL/gVS and 8.24±1.26 mL gVS^(-1)d^(-1)in the absence of NiFe_(2)O_(4)NPs(Group A)to 123.69±3.20 mL/gVS and 9.71±0.77 mL gVS^(-1)d^(-1)in the presence of NiFe_(2)O_(4)NPs(Group B).The model simulation results showed that both the first-order kinetic model and the modified Gompertz model can well simulate the experimental results.The hydrolysis rate constant k increased from 0.04±0.01 d^(-1)in Group A to 0.06±0.01 d^(-1)in Group B.And the maximum methane production potential and activity were both improved after adding NiFe_(2)O_(4).The microbial community analysis revealed that the microorganisms associated with hydrolysis and acidogenesis were more abundant in the presence of NiFe_(2)O_(4).And the methanogenic archaea were enriched to a larger extent,resulted in the higher methanogenesis activities via dosing NiFe_(2)O_(4).