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.

Direct Electron Transfer Reactions of Glucose Oxidase and D-Amino Acid Oxidase at a Glassy Carbon Electrode in Organic Media

Cyclic voltammetry is employed to demonstrate feasibility of direct electron transfer of glucose oxidase and D amino acid oxidase at a glassy carbon electrode in organic media. The reversible slight conformational change of glucose oxidase is observed by changing 0.1 mol/L phosphate buffer to acetonitrile containing 10% v/v of water and 0.05 mol/L tetrabutyalammonium perchlorate, and vice versa.

Electrically conductive nanowires controlled one pivotal route in energy harvest and microbial corrosion via direct metal-microbe electron transfer

Extracellular electron transfer(EET)plays a critical role in bioelectrochemical processes,allowing cou-pling between microorganisms and extracellular solid-state electrodes,metals,or other cells in energy metabolism.Previous studies have suggested a role for outer-surface c-type cytochromes in direct metal-to-microbe electron transfer by Geobacter sulfurreducens,a model electroactive bacterium.Here,we ex-amined the possibility of other microbially produced electrical contacts by deleting the gene for PilA,the protein monomer that G.sulfurreducens assembles into electrically conductive protein nanowires(e-pili).Deleting pilA gene inhibited electron extraction from pure iron and 316L stainless steel up to 31%and 81%,respectively more than deleting the gene for the outer-surface cytochrome OmcS.This PilA-deficient phenotype,and the observation that relatively thick biofilms(21.7μm)grew on the metal surfaces at multi-cell distances from the metal surfaces suggest that e-pili contributed significantly to microbial cor-rosion via direct metal-to-microbe electron transfer.These results have implications for the fundamental understanding of electron harvest via e-pili by electroactive microbes,their uses in bioenergy production,as well as in monitoring and mitigation of metal biocorrosion.

Direct Electron Transfer between Glucose Oxidase and Multi-walled Carbon Nanotubes

The direct electrochemical behavior between the glucose oxidase (GOD) and the multi walled carbon nanotubes (MWNTs) has been studied. Two pairs of cyclic voltammetric peaks corresponding to the two different processes, i.e. mass transport and surface reaction of GOD are observed on this MWNTs. The formal potentials with E o′=-0.45 V and E o′=-0.55 V were obtained respectively. The GOD film was observed on the carbon nanotube by the TEM.

Magnetic Fe_3O_4-Reduced Graphene Oxide Nanocomposites-Based Electrochemical Biosensing

An electrochemical biosensing platform was developed based on glucose oxidase(GOx)/Fe3O4-reduced graphene oxide(Fe3O4-RGO) nanosheets loaded on the magnetic glassy carbon electrode(MGCE).With the advantages of the magnetism, conductivity and biocompatibility of the Fe3O4-RGO nanosheets, the nanocomposites could be facilely adhered to the electrode surface by magnetically controllable assembling and beneficial to achieve the direct redox reactions and electrocatalytic behaviors of GOx immobilized into the nanocomposites. The biosensor exhibited good electrocatalytic activity, high sensitivity and stability. The current response is linear over glucose concentration ranging from 0.05 to 1.5 m M with a low detection limit of0.15 μM. Meanwhile, validation of the applicability of the biosensor was carried out by determining glucose in serum samples. The proposed protocol is simple, inexpensive and convenient, which shows great potential in biosensing application.

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.

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.

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.

Combining metal-microbe and microbe-microbe dual direct electron transfer on Fe(0)-cathode of bio-electrochemical system to enhance anaerobic digestion of cellulose wastewater

Considering that cathode of microbial electrochemical system(MES)is a good electrons source for methane production via direct/indirect electron transfer to electroactive microorganisms,and that Fe(0)is also a confirmed electron donor for some electroactive microorganisms through metal-microbe direct electron transfer(DET),Fe(0)-cathode was equipped into an MES digester to enhance cathodic methane production.The results of this study indicated that the potential DET participator,Clostridium possibly obtained electrons directly from Fe(0)-cathode via metal-microbe electrons transfer,then transferred electrons directly to the definite DET participators,Methanosarcina/Methanothrix via microbemicrobe electrons transfer for CH_(4)production.In addition,Methanobacterium is another specially enriched methanogen on Fe(0)-cathode,which might obtain electrons directly from Fe(0)-cathode to produce CH_(4) via metal/electrode-microbe DET.The increment of conductivity of cathodic sludge in Fe(0)-cathode MES digester(R1)further confirmed the enrichment of electroactive microorganisms participating in DET process.As a consequence,a higher CH_(4) production(1205–1508 m L/d)and chemical oxygen demand(COD)removal(79.0%-93.8%)were achieved in R1 compared with graphite-cathode MES digester(R2,720–1090 m L/d and 63.6%-85.6%)and the conventional anaerobic digester(R3,384–428 m L/d and 35.2%-41.0%).In addition,energy efficiency calculated indicated that the output energy of CH_(4) production was 8.16 folds of electricity input in Fe(0)-cathode MES digester.

A direct electrochemical biosensor for rapid glucose detection based on nitrogen-doped carbon nanocages

Given the increasing number of diabetic patients,rapid and accurate detection of glucose in body fluids is critical.This study developed a direct electrochemical biosensor for glucose based on nitrogen-doped carbon nanocages(NCNCs).NCNCs possess a large specific surface area of 1395 m^(2)·g^(-1),a high N atomic content of 9.37%and good biocompatibility,which is favorable for enzyme loading and electron transfer.The surface average concentration of electroactive glucose oxidase on NCNCs was 2.82×10^(-10)mol·cm^(-2).The NCNC-based direct electrochemical biosensor exhibited a high sensitivity of 13.7μA·(mmol·L^(-1))^(-1)·cm^(-2),rapid response time of 5 s and an impressive electron-transferrate constant(ks)of 1.87 s^(-1).Furthermore,we investigated an NCNC-based direct electron transfer(DET)biosensor for sweat glucose detection,which demonstrated tremendous promise for non-invasive wearable diabetes diagnosis.

Myoglobin/Gold Nanoparticles/Carbon Spheres 3-D Architecture for the Fabrication of a Novel Biosensor

A novel biosensor based on a myoglobin/gold nanoparticles/carbon spheres(Mb AuNPs CNs)3-D architecture bioconjunction has been fabricated for the determination of hydrogen peroxide(H2O2).Cyclic voltammetry(CV),Fourier transform infrared(FT-IR)spectroscopy and scanning electron microscopy(SEM)were used to characterize the bioconjunction of the AuNPs CNs with Mb.Experimental results demonstrate that the AuNPsCNs hybrid material is more effective in facilitating electron transfer of the immobilized enzyme than CNs alone,which can be attributed to the unique nanostructure and larger surface area of the bioconjunction.The biosensor displayed good performance for the detection of H_(2)O_(2)with a wide linear range from 0.28μmol/L to 116.5μmol/L and a detection limit of 0.12μmol/L.The Michaelis-Menten constant KMapp value was estimated to be 0.3 mmol/L.The resulting biosensor exhibited fast amperometric response,and good stability,reproducibility,and selectivity to H_(2)O_(2).

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).

Direct electrochemistry and electrocatalysis of myoglobin in dodecyltrimethylammonium bromide film modified carbon ceramic electrode

Direct electrochemistry and electrocatalysis of myoglobin（Mb） were studied with Mb immobilized on dodecyltrimethylammonium bromide（DTAB） film modified carbon ceramic（CC） electrode.Cyclic voltammetry showed a pair of well-defined and nearly reversible redox peaks of Mb（Fe~Ⅱ/Fe~Ⅲ） at about—0.3 V vs.SCE（pH = 6.98）.The currents of the redox peak were linear to scan rate,and rate constant（Ks） was estimated to be 3.03 s^（-1）.The formal potential（E°＇） of Mb in the DTAB/CC electrodes shifted linearly with pH with a slope of -36.44 mV/pH,implying that the electron transfer between DTAB and CC electrodes is accompanied by proton transportation.The immobilized Mb exhibited excellent electrocatalytic response to the reduction of hydrogen peroxide（H2O2）.

Immobilization of Hemoglobin on Cobalt Nanoparticles-modified Indium Tin Oxide Electrode： Direct Electrochemistry and Electrocatalytic Activity

A cobalt nanoparticles-attached indium tin oxide（CoNPs/ITO） electrode was applied to the immobilization of hemoglobin（Hb） and an Hb modified CoNPs/ITO electrode（Hb/CoNPs/ITO） was prepared. The direct electron transfer of Hb was shown by the well-behaved voltammetric responses for Hb/CoNPs/ITO electrode and the effects of scan rate and pH value were observed. Based on the catalytic activity of Hb immobilized on the CoNPs/ITO electrode toward the reduction of H2O2, a mediator-free H2O2 sensor was developed. A linear relationship existed between the catalytic current and the H2O2 concentration in a range of 1.0--100.0 μmol/L with a detection limit （S/N=3） of 0.2μmol/L.

Direct Electrochemistry of Hemoglobin on Vertically Aligned Carbon Hybrid TiO2 Nanotubes and Its Highly Sensitive Biosensor Performance

The present work is focused on developing a novel biomaterial platform to achieve enhanced direct electron transfer （DET） of hemoprotein and higher biosensor performance on vertically aligned carbon hybrid TiO2 nanotubes （C-TiO2 NTs）. Using a simple surfactant-assisted method, controllable hybridization of TiO2 NTs with conductive amorphous carbon species is realized. The obtained C-TiO2 NTs is ingeniously chosen to serve as an ideal ＂vessel＂ for protein immobilization and biosensor applications. Results show that the appropriate hybridization of C into TiO2 NTs leads to a much better conductivity of TiO2 NTs without destroying their preponderant tubular structures or damaging their excellent biocompatibility and hydrophilicity. When used in loading proteins, the C-TiO2 NTs can be used as a super vessel for rapid and substantive immobilization of hemoglobin （Hb）, with a large surface electroactive Hb coverage （I＊） of 3.3 × 10 9 mol·cm^-2. Enhanced DET of Hb is commendably observed on the constructed Hb/C-TiO2 NTs biosensor with a couple of well-defined redox peaks in a fast electron transfer process. The biosensor further exhibits fast response, high sensitivity and stability for the amperometric biosensing of H202 with the detection limit as low as 3.1 × 10^-8 mol/L.

Direct Electrochemistry of Hemoglobin Based on Fe3O4@SiO2 Nanoparticles Modified Electrode

A biosensor based on hemoglobin-Fe304@SiO2 nanoparticle bioconjunctions modified indium-tin-oxide （Hb/Fe3O4@SiOz/ITO） electrode was fabricated to determine the concentration of H202. UV-vis absorption spectra, fourier transform infrared （FT-IR） spectroscopy, cyclic voltammetry （CV） and high-resolution transmission electron microscopy （HRTEM） were used to characterize the bioconjunction of Fe3O4@SiO2 with Hb. Experimental results demonstrate that the immobilized Hb on the Fe3O4@SiO2 matrix retained its native structure well. In addition, Fe3O4@SiO2 nanoparticles （NPs） are very effective in facilitating electron transfer of the immobilized enzyme, which can be attributed to the unique nanostructure and larger surface area of the Fe304@SiO2 NPs. The biosensor displayed good performance for the detection of H2O2 with a wide linear range from 2.03×10^-6 to 4.05×10^3 mol/L and a detection limit of 0.32 μmol/L. The resulting biosensor exhibited fast amperometric response, good stability, reproducibility, and selectivity to H2O2.