Shin-ichi Hirano

Bioelectrochemical system stabilizes methane fermentation from garbage slurry.

Methanogenic bioreactors, which are packed with supporting material, have attracted attention as an efficient means of degrading garbage. We aimed to increase bioreactor performance by using an electrochemical system to regulate the electrical potential on supporting material. At an organic loading rate of 26.9g dichromate chemical oxygen demand (CODcr)/L/day, reactors with a potential of -0.6 or -0.8V, generated by a cathodic electrochemical reaction, showed greater removal of CODcr and methanogenesis than reactors with a potential of 0.0 or -0.3V, generated by anodic reaction, or control reactors without electrochemical regulation. 16S rRNA gene analysis revealed that the same methanogens were present in all our reactors, but quantitative real-time polymerase chain reaction showed that higher prokaryotic and methanogenic copy numbers were present on cathodic electrodes than on anodic or control electrodes. These results indicate that cathodic electrochemical regulation can support methane fermentation from garbage.

Effect of adding carbon fiber textiles to methanogenic bioreactors used to treat an artificial garbage slurry

To compare the performances and microbial populations of methanogenic reactors with and without carbon fiber textiles (CFT), we operated small-scale (200 ml) reactors using a slurry of artificial garbage. For both types of reactors, the organic loading rate (OLR) was stepwisely and rapidly increased in the same manner. Start-up period was shortened by adding CFT. Reactors with CFT showed greater efficiency for removal of suspended solid and volatile suspended solid than reactors without CFT at a long hydraulic retention time (HRT) between 8 and 13 days. The reactors with CFT maintained stable methane production at an OLR of 15.3 g dichromate chemical oxygen demand (CODcr)/l/day and DNAs from microorganisms were highly concentrated in adhering fractions on CFT. As shown by quantitative PCR analysis, the proportions of methanogenic archaea were conserved more than 25% in adhering fractions on CFT in reactors with CFT. By contrast, reactors without CFT showed accumulation of volatile fatty acid and deteriorated at an OLR of 2.4 gCODcr/l/day. Methanogenic proportions dropped to 17.1% in suspended fractions of reactors without CFT. Denaturing gradient gel electrophoresis (DGGE) analysis revealed that all archaeal DGGE bands in both types of reactors were related to methanogens, but more bands were observed in reactors with CFT. Thus the higher performance of reactors with CFT likely reflects the greater abundance of microorganisms and methanogenic diversity.

Electrochemical control of redox potential affects methanogenesis of the hydrogenotrophic methanogen Methanothermobacter thermautotrophicus.

To investigate the precise effect of the redox potential on the methanogenesis of the hydrogenotrophic methanogen Methanothermobacter thermautotrophicus by using an electrochemical redox controlling system without adding oxidizing or reducing agents. A bioelectrochemical system was applied to control the redox conditions in culture and to measure the methane‐producing activity of M. thermautotrophicus at a constant potential from +0·2 to −0·8 V (vs Ag/AgCl). Methane production and growth of M. thermautotrophicus were 1·6 and 3·5 times increased at −0·8 V, compared with control experiments without electrolysis, respectively, while methanogenesis was suppressed between +0·2 and −0·2 V. A clear relationship between an electrochemically regulated redox potential and methanogenesis was revealed.

A bioelectrochemical reactor containing carbon fiber textiles enables efficient methane fermentation from garbage slurry.

A packed-bed system includes supporting materials to retain microorganisms and a bioelectrochemical system influences the microbial metabolism. In our study, carbon fiber textiles (CFT) as a supporting material was attached onto a carbon working electrode in a bioelectrochemical reactor (BER) that degrades garbage slurry to methane, in order to investigate the effect of combining electrochemical regulation and packing CFT. The potential on the working electrode in the BER containing CFT was set to -1.0 V or -0.8 V (vs. Ag/AgCl). BERs containing CFT exhibited higher methane production, elimination of dichromate chemical oxygen demand, and the ratio of methanogens in the suspended fraction than reactors containing CFT without electrochemical regulation at an organic loading rate (OLR) of 27.8 gCODcr/L/day. In addition, BERs containing CFT exhibited higher reactor performances than BERs without CFT at this OLR. Our results revealed that the new design that combined electrochemical regulation and packing CFT was effective.

Methanogenic communities on the electrodes of bioelectrochemical reactors without membranes.

Methane fermentation was successfully carried out in bioelectrochemical reactors without membranes under a working potential of -0.6 or -0.8 V (vs. Ag/AgCl) and neutral pH conditions. The hydrogenotrophic methanogens that dominated on the anodic and cathodic electrodes differed from those found on the electrodes in the control reactors without electrochemical reactions.

Efficient treatment of garbage slurry in methanogenic bioreactor packed by fibrous sponge with high porosity

Adding a supporting material to a methanogenic bioreactor treating garbage slurry can improve efficiency of methane production. However, little is known on how characteristics (e.g., porosity and hydrophobicity) of the supporting material affect the bioreactor degrading garbage slurry. We describe the reactor performances and microbial communities in bioreactors containing hydrophilic or hydrophobic sheets, or fibrous hydrophilic or hydrophobic sponges. The porosity affected the efficiency of methane production and solid waste removal more than the hydrophilic or hydrophobic nature of the supporting material. When the terminal restriction fragment length polymorphism technique was used at a lower organic loading rate (OLR), microbial diversities in the suspended fraction were retained on the hydrophobic, but not the hydrophilic, sheets. Moreover, real-time quantitative polymerase chain reaction (PCR) performed at a higher OLR revealed that the excellent performance of reactors containing fibrous sponges with high porosity (98%) was supported by a clear increase in the numbers of methanogens on these sponges, resulting in larger total numbers of methanogens in the reactors. In addition, the bacterial communities in fractions retained on both the hydrophobic and hydrophilic fibrous sponges differed from those in the suspended fraction, thus increasing bacterial diversity in the reactor. Thus, higher porosity of the supporting material improves the bioreactor performance by increasing the amount of methanogens and bacterial diversity; surface hydrophobicity contributes to maintaining the suspended microbial community.

Bioelectrochemical regulation accelerates facultatively syntrophic proteolysis.

Bioelectrochemical systems can affect microbial metabolism by controlling the redox potential. We constructed bioelectrochemical cultures of the proteolytic bacterium, Coprothermobacter proteolyticus strain CT-1, both as a single-culture and as a co-culture with the hydrogenotrophic methanogen, Methanothermobacter thermautotrophicus strain ∆H, to investigate the influences of bioelectrochemical regulation on facultatively syntrophic proteolysis. The co-culture and single-culture were cultivated at 55°C with an anaerobic medium containing casein as the carbon source. The working electrode potential of the bioelectrochemical system was controlled at -0.8V (vs. Ag/AgCl) for bioelectrochemical cultures and was not controlled for non-bioelectrochemical cultures. The cell densities of hydrogenotrophic methanogen and methane production in the bioelectrochemical co-culture were 3.6 and 1.5 times higher than those in the non-bioelectrochemical co-culture after 7 days of cultivation, respectively. Contrastingly, the cell density of Coprothermobacter sp. in the bioelectrochemical co-culture was only 1.3 times higher than that in the non-bioelectrochemical co-culture. The protein decomposition rates were nearly proportional to the cell density of Coprothermobacter sp. in the all types of cultures. These results indicate that bioelectrochemical regulation, particularly, affected the carbon fixation of the hydrogenotrophic methanogen and that facultatively syntrophic proteolysis was accelerated as a result of hydrogen consumption by the methanogens growing well in bioelectrochemical co-cultures.

Electrical-biological hybrid system for CO2 reduction

Here we have developed an electrochemical-biological hybrid system to fix CO2. Natural biological CO2 fixation processes are relatively slow. To increase the speed of fixation we applied electrocatalysts to reduce CO2 to formate. We chose a user-friendly organism, Escherichia coli, as host. Overall, the newly constructed CO2 and formate fixation pathway converts two formate and one CO2 to one pyruvate via glycine and L-serine in E. coli. First, one formate and one CO2 are converted to one glycine. Second, L-serine is produced from one glycine and one formate. Lastly, L-serine is converted to pyruvate. E. colis genetic tractability allowed us to balance various parameters of the pathway. The carbon flux of the pathway was sufficient to compensate L-serine auxotrophy in the strain. In total, we integrated both electrocatalysis and biological systems into a single pot to support E. coli growth with CO2 and electricity. Results show promise for using this hybrid system for chemical production from CO2 and electricity.

Analysis of a bio-electrochemical reactor containing carbon fiber textiles for the anaerobic digestion of tomato plant residues

A bio-electrochemical system packed with supporting material can promote anaerobic digestion for several types of organic waste. To expand the target organic matters of a BES, tomato plant residues (TPRs), generated year-round as agricultural and cellulosic waste, were treated using three methanogenic reactors: a continuous stirred tank reactor (CSTR), a carbon fiber textile (CFT) reactor, and a bio-electrochemical reactor (BER) including CFT with electrochemical regulation (BER + CFT). CFT had positive effects on methane fermentation and methanogen abundance. The microbial population stimulated by electrochemical regulation, including hydrogenotrophic methanogens, cellulose-degrading bacteria, and acetate-degrading bacteria, suppressed acetate accumulation, as evidenced by the low acetate concentration in the suspended fraction in the BER + CFT. These results indicated that the microbial community in the BER + CFT facilitated the efficient decomposition of TPR and its intermediates such as acetate to methane.

Enrichment of Chromium-Reducing Bacteria Using Potential-Controlled Electrochemical Cultivation

We present here a method for the electrochemical enrichment of chromium-reducing bacteria from environmental samples containing various microorganisms. An electrochemical cultivation system was constructed; this consisted of a Pt anode and a carbon cathode separated by an ion-exchange membrane. For an electrochemical cultivation, an environmental sample with chromium-reducing bacteria was inoculated into the anode well where a medium containing 0.1 mM Cr(VI) ions was poured. On the other hand, Cr(VI)-free medium was poured into the cathode well. Chromium-reducing bacteria grew selectively after 10 days of incubation and application of 1.0 V (vs. Ag/AgCl) of anode potential that reproduces Cr(VI) ions. This result suggested that Cr(VI) ions generated on the anode promoted the growth of chromium-reducing bacteria as their electron acceptor and due to Cr(VI) toxicity, inhibited the growth of other microorganisms.