Enrichment of homoacetogens converting H2/CO2 into acids and ethanol and simultaneous methane production.

An anaerobic granular sludge was enriched to utilize H2/CO2 in a continuous gas-fed up-flow anaerobic sludge reactor by applying operating conditions expected to produce acetic acid, butyric acid, and ethanol. Three stages of fermentation were found: Stage I with acetic acid accumulation with the highest concentration of 35 mM along with a pH decrease from initial 6 to 4.5. In Stage II, H2/CO2 was replaced by 100% H2 to induce solventogenesis, whereas butyric acid was produced with the highest concentration of 2.5 mM. At stage III with 10 µM tungsten (W) addition, iso-valeric acid, valeric acid, and caproic acid were produced at pH 4.5-5.0. In the batch tests inoculated with the enriched sludge taken from the bioreactor (day 70), however, methane production occurred at pH 6. Exogenous 15 mM acetate addition enhanced both the H2 and CO2 consumption rate compared to exogenous 10, 30, and 45 mM acetate by the enriched sludge. Exogenous acetate was failed to be converted to ethanol using H2 as electron donor by the enriched acetogens.

Enhanced removal of mixed VOCs with different hydrophobicities by Tween 20 in a biotrickling filter: Kinetic analysis and biofilm characteristics.

Mass transfer limitation usually causes the poor performance of biotrickling filters (BTFs) for the treatment of hydrophobic volatile organic compounds (VOCs) during long-term operation. In this study, two identical lab-scale BTFs were established to remove a mixture of n-hexane and dichloromethane (DCM) gases using non-ionic surfactant Tween 20 by Pseudomonas mendocina NX-1 and Methylobacterium rhodesianum H13. A low pressure drop (≤110 Pa) and a rapid biomass accumulation (17.1 mg g-1) were observed in the presence of Tween 20 during the startup period (30 d). The removal efficiency (RE) of n-hexane was enhanced by 15.0%- 20.5% while DCM was completely removed with the inlet concentration (IC) of 300 mg·m-3 at different empty bed residence times in the Tween 20 added BTF. The viable cells and the relative hydrophobicity of the biofilm were increased under the action of Tween 20, which facilitated the mass transfer and enhanced the metabolic utilization of pollutants by microbes. Besides, Tween 20 addition enhanced the biofilm formation processes including the increased extracellular polymeric substance (EPS) secretion, biofilm roughness and biofilm adhesion. The kinetic model simulated the removal performance of the BTF with Tween 20 for the mixed hydrophobic VOCs, and the goodness-of-fit was above 0.9.

Biofilm metagenomic characteristics behind high coulombic efficiency for propanethiol deodorization in two-phase partitioning microbial fuel cell.

Hydrophobic volatile organic sulfur compounds (VOSCs) are frequently found during sewage treatment, and their effective management is crucial for reducing malodorous complaints. Microbial fuel cells (MFC) are effective for both VOSCs abatement and energy recovery. However, the performance of MFC on VOSCs remains limited by the mass transfer efficiency of MFC in aqueous media. Inspired by two-phase partitioning biotechnology, silicone oil was introduced for the first time into MFC as a non-aqueous phase (NAP) medium to construct two-phase partitioning microbial fuel cell (TPPMFC) and augment the mass transfer of target VOSCs of propanethiol (PT) in the liquid phase. The PT removal efficiency within 32 h increased by 11-20% compared with that of single-phase MFC, and the coulombic efficiency of TPPMFC (11.01%) was 4.32-2.68 times that of single-phase MFC owing to the fact that highly active desulfurization and thiol-degrading bacteria (e.g., Pseudomonas, Achromobacter) were attached to the silicone oil surface, whereas sulfur-oxidizing bacteria (e.g., Thiobacillus, Commonas, Ottowia) were dominant on the anodic biofilm. The outer membrane cytochrome-c content and NADH dehydrogenase activity improved by 4.15 and 3.36 times in the TPPMFC, respectively. The results of metagenomics by KEGG and COG confirmed that the metabolism of PT in TPPMFC was comprehensive, and that the addition of a NAP upregulates the expression of genes related to sulfur metabolism, energy generation, and amino acid synthesis. This finding indicates that the NAP assisted bioelectrochemical systems would be promising to solve mass-transfer restrictions in low solubility contaminates removal.

Enhanced solventogenesis in syngas bioconversion: Role of process parameters and thermodynamics.

Biofuels, such as ethanol and butanol, obtained from carbon monoxide-rich gas or syngas bioconversion (solventogenesis) are an attractive alternative to traditional fermentation processes with merits of no competition with food production and sustainability. However, there is a lack of comprehensive understanding of some key process parameters and mechanisms enhancing solventogenesis during the fermentation process. This review provides an overview of the current state of the art of the main influencing factors during the syngas fermentation process catalyzed by acetogenic species as well as undefined mixed cultures. The role of syngas pressure, syngas components, fermentation pH, temperature, trace metals, organic compounds and additional materials is overviewed. As a so far hardly considered approach, thermodynamic calculations of the Gibbs free energy of CO conversion to acetic acid, ethanol, butyric acid and butanol under different CO pressures and pH at 25, 33 and 55 °C are also addressed and reviewed. Strategies for enhancing mass transfer and longer carbon chain solvent production are considered as well.

Effect of Endogenous and Exogenous Butyric Acid on Butanol Production From CO by Enriched Clostridia.

Butanol is a potential renewable fuel. To increase the selectivity for butanol during CO fermentation, exogenous acetic acid and ethanol, exogenous butyric acid or endogenous butyric acid from glucose fermentation have been investigated using CO as reducing power, with a highly enriched Clostridium sludge. Addition of 3.2 g/L exogenous butyric acid led to the highest 1.9 g/L butanol concentration with a conversion efficiency of 67%. With exogenous acetate and ethanol supply, the butanol concentration reached 1.6 g/L at the end of the incubation. However, the presence of acetic acid and ethanol favoured butanol production to 2.6 g/L from exogenous butyric acid by the enriched sludge. Finally, exogenous 14 g/L butyric acid yielded the highest butanol production of 3.4 g/L, which was also among the highest butanol concentration from CO/syngas fermentation reported so far. CO addition triggered butanol production from endogenous butyric acid (produced from glucose, Glucose + N2) with as high as 58.6% conversion efficiency and 62.1% butanol yield. However, no efficient butanol production was found from glucose and CO co-fermentation (Glucose + CO), although a similar amount of endogenous butyric acid was produced compared to Glucose + N2. The Clostridium genus occupied a relative abundance as high as 82% from the initial inoculum, while the Clostridia and Bacilli classes were both enriched and dominated in Glucose + N2 and Glucose + CO incubations. This study shows that the supply of butyric acid is a possible strategy for enhancing butanol production by CO fed anaerobic sludge, either via exogenous butyric acid, or via endogenous production by sugar fermentation.

Selective butanol production from carbon monoxide by an enriched anaerobic culture.

An anaerobic mixed culture able to grow on pure carbon monoxide (CO) as well as syngas (CO, CO2 and H2), that produced unusual high concentrations of butanol, was enriched in a bioreactor with intermittent CO gas feeding. At pH 6.2, it mainly produced acids, generally acetic and butyric acid. After adaptation, under stress conditions of CO exposure at a partial pressure of 1.8 bar and low pH (e.g., 5.7), the enrichment accumulated ethanol, but also high amounts of butanol, up to 6.8 g/L, never reported before, with a high butanol/butyric acid molar ratio of 12.6, highlighting the high level of acid to alcohol conversion. At the end of the assay, both the acetic acid and ethanol concentrations decreased, with concomitant butyric acid production, suggesting C2 to C4 acid bioconversion, though this was not a dominant bioconversion process. The reverse reaction of ethanol oxidation to acetic acid was observed in the presence of CO2 produced during CO fermentation. Interestingly, butanol oxidation with simultaneous butyric acid production occurred upon production of CO2 from CO, which has to the best of our knowledge never been reported. Although the sludge inoculum contained a few known solventogenic Clostridia, the relative taxonomic abundance of the enriched sludge was diverse in Clostridia and Bacilli classes, containing known solventogens, e.g., Clostridium ljungdhalii, Clostridium ragsdalei and Clostridium coskatii, confirming their efficient enrichment. The relative abundance of unassigned Clostridium species amounted to 27% with presumably novel ethanol/butanol producers.

Bioethanol Production From H2/CO2 by Solventogenesis Using Anaerobic Granular Sludge: Effect of Process Parameters.

CO2 fermentation by biocatalysis is a promising route for the sustainable production of valuable chemicals and fuels, such as acetic acid and ethanol. Considering the important role of environmental parameters on fermentation processes, granular sludge from an industrial anaerobic wastewater treatment system was tested as inoculum for ethanol production from H2/CO2 at psychrophilic (18°C), submesophilic (25°C), and mesophilic (30°C) temperatures. The highest acetic acid and ethanol production was obtained at 25°C with a final concentration of 29.7 and 8.8 mM, respectively. The presence of bicarbonate enhanced acetic acid production 3.0 ∼ 4.1-fold, while inhibiting ethanol production. The addition of 0.3 g/L glucose induced butyric acid production (3.7 mM), while 5.7 mM ethanol was produced at the end of the incubation at pH 4 with glucose. The addition of 10 µM W enhanced ethanol production up to 3.8 and 7.0-fold compared to, respectively, 2 µM W addition and the control. The addition of 2 µM Mo enhanced ethanol production up to 8.1- and 5.4-fold compared to, respectively, 10 µM Mo and the control. This study showed that ethanol production from H2/CO2 conversion using granular sludge as the inoculum can be optimized by selecting the operational temperature and by trace metal addition.

Enhanced Ethanol Production From Carbon Monoxide by Enriched Clostridium Bacteria.

Carbon monoxide (CO)-metabolizing Clostridium spp. were enriched from the biomass of a butanol-producing reactor. After six successive biomass transfers, ethanol production reached as much as 11.8 g/L with minor accumulation of acetic acid, under intermittent gas feeding conditions and over a wide pH range of 6.45-4.95. The molar ratio of ethanol to acetic acid exceeded 1.7 after the lag phase of 11 days and reached its highest value of 8.6 during the fermentation process after 25 days. Although butanol production was not significantly enhanced in the enrichment, the biomass was able to convert exogenous butyric acid (3.2 g/L) into butanol with nearly 100% conversion efficiency using CO as reducing power. This suggested that inhibition of butanol production from CO was caused by the lack of natural butyric acid production, expectedly induced by unsuitable pH values due to initial acidification resulting from the acetic acid production. The enriched Clostridium population also converted glucose to formic, acetic, propionic, and butyric acids in batch tests with daily pH adjustment to pH 6.0. The Clostridium genus was enriched with its relative abundance significantly increasing from 7% in the inoculum to 94% after five successive enrichment steps. Unidentified Clostridium species showed a very high relative abundance, reaching 73% of the Clostridium genus in the enriched sludge (6th transfer).

Homoacetogenesis and solventogenesis from H2/CO2 by granular sludge at 25, 37 and 55 °C.

CO2 fermentation is a promising process to produce biofuels like ethanol. It can be integrated in third generation biofuel production processes to substitute traditional sugar fermentation when supplied with cheap electron donors, e.g. hydrogen derived from wind energy or as surplus gas in electrolysis. In this study, granular sludge from an industrial wastewater treatment plant was tested as inoculum for ethanol production from H2/CO2 via non-phototropic fermentation at submesophilic (25 °C), mesophilic (37 °C) and thermophilic (55 °C) conditions. The highest ethanol concentration (17.11 mM) was obtained at 25 °C and was 5-fold higher than at 37 °C (3.36 mM), which was attributed to the fact that the undissociated acid (non-ionized acetic acid) accumulation rate constant (0.145 h-1) was 1.39 fold higher than at 25 °C (0.104 h-1). Methane was mainly produced at 55 °C, while neither acetic acid nor ethanol were formed. Ethanol production was linked to acetic acid production with the highest ethanol to acetic acid ratio of 0.514 at 25 °C. The carbon recovery was 115.7%, 131.2% and 117.1%, while the electron balance was almost closed (97.1%, 110.1% and 109.1%) at 25 °C, 37 °C and 55 °C, respectively. The addition of bicarbonate inhibited ethanol production both at 25 °C and 37 °C. Clostridium sp. were the prevalent species at both 25 and 37 °C at the end of the incubation, which possibly contributed to the ethanol production.

Bioremediation of Cr (VI) contaminated groundwater by Geobacter sulfurreducens: Environmental factors and electron transfer flow studies.

In this study, the removal of Cr (VI) was examined in the presence of bio-produced Fe (II) from hematite, sulfate and dissolved organic matter by Geobacter sulfurreducens. The adaptation results of G. sulfurreducens showed that cells growth was stimulated up to 576 µM of Cr (VI) concentration. The first-order rate and electron transfer rate in each step during Cr (VI) reduction by G. sulfurreducens in the presence of hematite was clearly modeled and calculated. For Cr (VI) reduction rate, both separately dissolved and adsorbed bio-produced Fe (II) were faster than G. sulfurreducens although bio-produced Fe (II) contributed only 20% to total Cr (VI) removal in a combined system containing Cr (VI), hematite and G. sulfurreducens. The electron transfer rate from G. sulfurreducens to hematite (R2) to produce Fe (II) was a limited step and electron transfer rate from acetate to Cr (VI) (1.8 µeq L-1 h-1) by G. sulfurreducens was much higher than that to hematite (0.272 µeq L-1 h-1, producing Fe (II)). Cr (VI) reduction was enhanced in the presence of SO42- due to sulfate boost cells growth. AQDS enhanced Cr (VI) reduction by serving as an electron shuttle thus accelerating the electron transfer rate.

Intracellular versus extracellular accumulation of Hexavalent chromium reduction products by Geobacter sulfurreducens PCA.

Hexavalent chromium (Cr(VI)) reduction by Geobacter sulfurreducens PCA was evaluated in batch experiments, and the form and amounts of intracellular and extra-cellular Cr(VI) reduction products were determined over time. The first-order Cr(VI) reduction rate per unit mass of cells was consistent for different initial cell concentrations, and approximately equal to (2.065 ±â€¯0.389) x 10-9 mL CFU-1 h-1. A portion of the reduced Cr(VI) products precipitated on Geobacter cell walls as Cr(III) and was bound via carboxylate functional groups, a portion accumulated inside Geobacter cells, and another portion existed as soluble Cr(III) or organo-Cr(III) released to solution. A mass balance analysis of total chromium in aqueous media, on cell walls, and inside cells was determined as a function of time, and with different initial cell concentrations. Mass balances were between 92% and 98%, and indicated Cr(VI) reduction products accumulate more on cell walls and inside cells with time and with increasing initial cell concentration, as opposed to particulates in aqueous solution. Reduced Cr(VI) products both in solution and on cell surfaces appear to form organo-Cr(III) complexes, and our results suggest that such complexes are more stable to reoxidation than aqueous Cr(III) or Cr(OH)3. Chromium inside cells is also likely more stable to reoxidation, both because it can form organic complexes, and it is separated by the cell membrane from solution conditions. Hence, Cr(VI) reduction products in groundwater during bioremediation may become more stable against re-oxidation, and may pose a lower risk to human health, over time and with greater initial biomass densities.