Kun Dai

Stable acetate production in extreme-thermophilic (70°C) mixed culture fermentation by selective enrichment of hydrogenotrophic methanogens

The control of metabolite production is difficult in mixed culture fermentation. This is particularly related to hydrogen inhibition. In this work, hydrogenotrophic methanogens were selectively enriched to reduce the hydrogen partial pressure and to realize efficient acetate production in extreme-thermophilic (70°C) mixed culture fermentation. The continuous stirred tank reactor (CSTR) was stable operated during 100 days, in which acetate accounted for more than 90% of metabolites in liquid solutions. The yields of acetate, methane and biomass in CSTR were 1.5 ± 0.06, 1.0 ± 0.13 and 0.4 ± 0.05 mol/mol glucose, respectively, close to the theoretical expected values. The CSTR effluent was stable and no further conversion occurred when incubated for 14 days in a batch reactor. In fed-batch experiments, acetate could be produced up to 34.4 g/L, significantly higher than observed in common hydrogen producing fermentations. Acetate also accounted for more than 90% of soluble products formed in these fed-batch fermentations. The microbial community analysis revealed hydrogenotrophic methanogens (mainly Methanothermobacter thermautotrophicus and Methanobacterium thermoaggregans) as 98% of Archaea, confirming that high temperature will select hydrogenotrophic methanogens over aceticlastic methanogens effectively. This work demonstrated a potential application to effectively produce acetate as a value chemical and methane as an energy gas together via mixed culture fermentation.

Electricity production and microbial characterization of thermophilic microbial fuel cells

Thermophilic microbial fuel cell (TMFC) offers many benefits, but the investigations on the diversity of exoelectrogenic bacteria are scarce. In this study, a two-chamber TMFC was constructed using ethanol as an electron donor, and the microbial dynamics were analyzed by high-throughput sequencing and 16S rRNA clone-library sequencing. The open-circuit potential of TMFC was approximately 650mV, while the maximum voltage was around 550mV. The maximum power density was 437mW/m2, and the columbic efficiency in this work was 20.5±6.0%. The Firmicutes bacteria, related to the uncultured bacterium clone A55_D21_H_B_C01 with a similarity of 99%, accounted for 90.9% of all bacteria in the TMFC biofilm. This unknown bacterium has the potential to become a new thermophilic exoelectrogenic bacterium that is yet to be cultured. The development of TMFC-involved biotechnologies will be beneficial for the production of valuable chemicals and generation of energy in the future.

Valuable biochemical production in mixed culture fermentation: fundamentals and process coupling

The mixed culture fermentation is an important environmental biotechnology that converts biodegradable organic wastes to valuable chemicals such as hydrogen, methane, acetate, ethanol, propionate, and so on. For the multistep process of hydrolysis, acidogenesis, acetogenesis/homoacetogensis, and methanogenesis, the typical metabolic reactions are firstly summarized. And then, since the final metabolites are always a mixture, the separation and purification processes are necessary to couple with anaerobic fermentation. Therefore, several typical coupling technologies including biogas upgrading, two-stage fermentation, gas stripping, membrane technology of pervaporation, membrane distillation, electrodialysis, bipolar membrane electrodialysis, and microbial fuel cells are summarized to separate the metabolites and recover energy. At last, the novel technologies such as the controlled metabolite production, medium chain carboxylic acid production, and high temperature ethanol recovery in thermophilic mixed culture fermentation are also reviewed. However, the novel concepts are still needed to meet the demands of better overall performances and lower total costs.

Hydrogen and carbon dioxide mixed culture fermentation in a hollow-fiber membrane biofilm reactor at 25 °C

There have been no reports of H2 and CO2 mixed-culture fermentation (MCF) at 25 °C in a hollow-fiber membrane biofilm reactor (HfMBR). In this study, H2 and CO2 MCF were conducted in an HfMBR at 25 °C producing metabolites including acetate, ethanol, butyrate, and caproate. Compared to pure culture fermentation (i.e., Clostridium carboxidivorans P7), the MCF in HfMBR at 25 °C produced a higher concentration of caproate in this study (3.4 g/L in batch 1 and 5.7 g/L in batch 2). The dominant genera were Clostridium_sensu_stricto_12 and Prevotella_7. The caproate was more likely formed from the pathway of acetate and ethanol rather than via butyrate and ethanol. Since caproate is more valuable than acetate and low temperature fermentation consumes less energy, this process of H2 and CO2 MCF at 25 °C is appropriate for industrial application.

Free acetic acid as the key factor for the inhibition of hydrogenotrophic methanogenesis in mesophilic mixed culture fermentation

The inhibition of acetate under acidic pH is an ideal way to reduce methanogenesis in mesophilic mixed culture fermentation (MCF). However, the effects of acetate concentration and acidic pH on methanogenesis remain unclear. Besides, although hydrogenotrophic methanogens can be suitable targets in MCF, they are generally ignored. Therefore, we intentionally enriched hydrogenotrophic methanogens and found that free acetic acid (FAA, x) concentration and specific methanogenic activity (SMA, y) were correlated according to the equation: y = 0.86 × 0.31/(0.31 + x) (R2 = 0.909). The SMA was decreased by 50% and 90% at the FAA concentrations of 0.31 and 2.36 g/L, respectively. The coenzyme M concentration and relative electron transport activity agreed well with the FAA concentration. Moreover, the methanogenic activity could not be recovered when the FAA concentration exceeded 0.81 g/L. These findings indicated that neither acetate nor acidic pH, but FAA was the key factor to inhibit methanogenesis in MCF.

The chemostat metabolite spectra of alkaline mixed culture fermentation under mesophilic, thermophilic, and extreme-thermophilic conditions

Alkaline mixed culture fermentation (MCF) is a promising technology for reducing organic waste and producing biochemicals. However, chemostat metabolite spectra that are necessary for constructing a model and analyzing factors are seldom reported. In the present study, the effects of pH on the metabolites distribution in mesophilic (35 °C), thermophilic (55 °C), and extreme-thermophilic (70 °C) alkaline MCF were demonstrated. A chemical oxygen demand balance above 80% was achieved, and the main metabolites included acetate, ethanol, propionate, lactate, and formate. The yields of ethanol and formate increased as pH was increased from 7.5 to higher pH under mesophilic and thermophilic conditions, while the yields of acetate, lactate, and/or propionate decreased. The yields of ethanol, acetate, and formate increased under extreme-thermophilic conditions as pH was increased from 7.5 to 9.0, whereas lactate and hydrogen yields decreased. Low biomass yield under thermophilic and extreme-thermophilic conditions benefited higher metabolite production and minimized the accumulation of sludge.