Hydrogen-producing Reactors Research Articles

Work scheme to isolate the different micro-organisms found in hydrogen-producing reactors: a study of effectiveness by pyrosequencing analysis.

The aim of this research was to create a work scheme for the isolation of the different micro-organisms commonly found in hydrogen-producing reactors and to test its effectiveness. Methods were selected to isolate anaerobic spore-forming fermenters, anaerobic fermenters that do not form spores, facultative aerobic fermenters and lactic acid bacteria. The methods were tested in two samples taken from a hydrogen-producing reactor fed with cheese whey. 16S rRNA gene sequences from isolates were compared with pyrosequencing analysis from the same samples. The isolates represented more than 88% of the abundance detected by pyrosequencing. Organisms from the genera Clostridium, Rahnella, Megasphaera, Lactobacillus, Propionibacterium, Bifidobacterium, Chryseobacterium and Acetobacter were isolated. Hydrogen-producing capacity was confirmed for the Clostridium, Rahnella and Megasphaera isolates. Coculture experiments indicate that Megasphaera prevented the total inhibition of Clostridium by Lactobacillus. The work scheme proposed was effective to isolate most of the micro-organisms detected by pyrosequencing analysis. Physiological studies suggested a key role of Megasphaera. We showed the high culturability of the microbial communities from hydrogen-producing bioreactors. The isolates can be used to perform physiological studies to understand the H2 -producing process.

Biomethane generation in an AnSBBR treating effluent from the biohydrogen production from vinasse: Optimization, metabolic pathways modeling and scale-up estimation

Abstract A study was performed regarding the production of methane by an AnSBBR treating wastewater coming from a biohydrogen production process using vinasse (sugar cane stillage). The reactor operated with recirculation of liquid phase in sequencing batch and fed-batch mode. The influence of the applied volumetric organic load (AVOL) was examined by varying influent concentration between 1000 and 4500 mgCOD L−1 (1.5–6.6 kgCOD m−3 d−1). Increasing AVOL resulted in a decrease in organic matter removal efficiency and in an increase in methane productivity. The highest productivity (133.1 molCH4 m−3 d−1) was obtained in fed-batch operation at the highest AVOL. Methane productivity and yield were always higher for fed-batch operation. From the kinetic metabolic model, it was possible to infer that methane is mainly produced via the acetoclastic route in batch mode and via both acetoclastic and hydrogenotrophic routes in fed-batch mode. Energy production of the two-stage system (acidogenic/biohydrogen-methanogenic/methane) was 13.6 kJ per gram of applied COD, which corresponds to a 38.8% increase compared to the traditional one-stage system (methanogenic/methane). The scale-up assessment (based on industrial production and performed with best condition data) proposed an operation of six parallel hydrogen-producing reactors of 6076 m3 each followed by four parallel methane-producing reactors of 1720 m3 each.

Microbial communities from 20 different hydrogen-producing reactors studied by 454 pyrosequencing.

To provide new insight into the dark fermentation process, a multi-lateral study was performed to study the microbiology of 20 different lab-scale bioreactors operated in four different countries (Brazil, Chile, Mexico, and Uruguay). Samples (29) were collected from bioreactors with different configurations, operation conditions, and performances. The microbial communities were analyzed using 16S rRNA genes 454 pyrosequencing. The results showed notably uneven communities with a high predominance of a particular genus. The phylum Firmicutes predominated in most of the samples, but the phyla Thermotogae or Proteobacteria dominated in a few samples. Genera from three physiological groups were detected: high-yield hydrogen producers (Clostridium, Kosmotoga, Enterobacter), fermenters with low-hydrogen yield (mostly from Veillonelaceae), and competitors (Lactobacillus). Inocula, reactor configurations, and substrates influence the microbial communities. This is the first joint effort that evaluates hydrogen-producing reactors and operational conditions from different countries and contributes to understand the dark fermentation process.

Role of homo-and heterofermentative lactic acid bacteria on hydrogen-producing reactors operated with cheese whey wastewater

The prevalence of lactic acid bacteria (LAB) and the effects of their antimicrobial peptides on H2 production in anaerobic fluidized bed reactors (AFBRs) operated with cheese whey (AFBR1 and AFBR2) were verified in this study. The AFBR1 received 5 g COD L−1 of cheese whey with decreasing hydraulic retention times (HRT) of 14 to 8 h. The AFBR2 was operated with 3–10 g COD L−1 of cheese whey with an HRT of 6 h. Next, 152 colonies were selected from de Man, Rogosa and Sharpe MRS agar plates, and 45 strains were classified as LAB. The counts oscillated between 6.6 and 8.1 log CFU mL−1, indicating that the LAB survived and persisted in the AFBRs. Pure cultures were identified using 16S rRNA gene sequencing, and Lactococcus lactis was the prevalent LAB (70%) in both reactors. The highest H2 yields (1.9 and 2.3 mol H2 mol lactose−1) were obtained during the first operational phase in both reactors when a low organic loading rate (OLR) was applied, and when the growth of Lactococcus spp. was associated with Leuconostoc pseudomesenteroides. The bacteriocin-producing LAB (mostly Lactobacillus spp.) found on the specific phases of reactors AFBR1 and AFBR2 exerted a remarkable influence on H2 yield.

Strategies to cope with methanogens in hydrogen producing UASB reactors: Community dynamics

Methane occurrence is a common concern in hydrogen producing reactors. This study presents the analysis of the microbial community structure during the application of operational strategies to decrease methane production, in three different up-flow anaerobic sludge blanket hydrogen-producing reactors. Cloning and denaturing gradient gel electrophoresis approach were used to establish the presence of homoacetogens, methanogens and hydrogen producers. The results showed that homoacetogenic organisms related to Blautia hydrogenotrophica and Oscillibacter valericigenes, and the hydrogen producer Enterobacter aerogenes where favored during pH decreasing strategies (5.6 to 4.5). The increment of the organic loading rate from 20 to 30 g chemical oxygen demand/L-d, selected hydrogen producers similar to Clostridium tyrobutyricum, Citrobacter freundii and E. aerogenes; further increments caused inhibition of hydrogen production due to the high undissociated acids concentration. Methane production was inhibited completely only when the biomass of the reactor was heat treated for a second time, this strategy selected hydrogen producers capable to sporulate, but homoacetogens were also favored. In all reactors the methanogenic activity was attributed to hydrogenotrophs related to the genera Methanobrevibacter and Methanobacterium.

Decreasing methane production in hydrogenogenic UASB reactors fed with cheese whey

One of the problems in fermentative hydrogen producing reactors, inoculated with pre-treated anaerobic granular sludge, is the eventual methane production by hydrogen-consuming methanogens. In this study, strategies such as reduction of pH and HRT, organic shock loads and repeated biomass heat treatment were applied to hydrogenogenic UASB reactors fed with cheese whey, that showed methane production after certain time of continuous operation (between 10 and 60 days). The reduction of pH to 4.5 not only decreased methane production but also hydrogen production. Organic shock load (from 20 to 30 g COD/L-d) was the more effective strategy to decrease the methane production rate (75%) and to increase the hydrogen production rate (172%), without stopping reactor operation. Repeated heat treatment of the granular sludge was the only strategy that inhibited completely methane production, leading to high volumetric hydrogen production rates (1.67 L H2/L-d), however this strategy required stopping reactor operation; in addition homoacetogenesis, another hydrogen-consuming pathway, was not completely inhibited. This work demonstrated that it was possible to control the methane activity in hydrogen producing reactors using operational strategies.

Enhanced hydrogen production in a UASB reactor by retaining microbial consortium onto carbon nanotubes (CNTs)

Biohydrogen and subsequent biomethane generation from biomass is a promising strategy for renewable energy supply, because this combination can lead to higher energy recovery efficiency and faster fermentation than single methane fermentation. Microbial consortium control by retaining hydrogen-producers through the addition of microbial carriers is an alternative to constructing hydrogen-producing reactors. Here we report the use of carbon nanotubes (CNTs) as microbial carriers to enhance microbial retention and the production of biohydrogen. Laboratory-scale upflow anaerobic sludge blanket (UASB) reactors with CNTs at 100 mg/L achieved a maximal hydrogen production rate of 5.55 L/L/d and a maximal hydrogen yield of 2.45 mol/mol glucose. Compared to frequently used activated carbon (AC) particles, CNTs resulted in quicker startup and better performance of hydrogen fermentation in UASB reactors. Scanning electron microscopy (SEM) and pyrosequencing results revealed that the reactor with CNTs led to a high proportion of hydrogen-producing bacteria among the microbial consortium, which endowed the microbes with strong flocculation capacity and hydrogen productivity.

Harnessing Bacteria That Use Light To Produce Hydrogen

Rhodopseudomonas palustris is one of several types of microorganisms that produce hydrogen gas (H2), a deceptively simple molecule consisting of two protons and two electrons. We study how R. palustris produces H2 in part because this organism so aptly illustrates the different types of data and skill sets that are required to bring a microbial metabolic property into practical use. Focused studies on specific mechanisms, broader approaches to decipher extensive biological networks, and nonbiological skills such as those needed to design and operate hydrogen-producing reactors all come into play as part of an effort to harness microbes to augment and replace unsustainable petrochemical processes.

16S rRNA-targeted probes for specific detection of Thermoanaerobacterium spp., Thermoanaerobacterium thermosaccharolyticum, and Caldicellulosiruptor spp. by fluorescent in situ hybridization in biohydrogen producing systems

16S rRNA gene targeted oligonucleotide probes for specific detection of genera Thermoanaerobacterium (Tbm1282), Caldicellulosiruptor (Ccs432), and specie Thermoanaerobacterium thermosaccharolyticum (Tbmthsacc184) were designed and used to monitor the spatial distribution of hydrogen producing bacteria in sludge and granules from anaerobic reactors. The designed probes were checked for their specificity and then validated using fluorescence in situ hybridization with target microorganisms and non-target microorganisms. Thermoanaerobacterium spp., T. thermosaccharolyticum and Caldicellulosiruptor spp. were detected with the probes designed with coverage of 75%, 100% and 93%, respectively. Thermophilic (60 °C) hydrogen producing reactors, one fed with sucrose and another, fed with palm oil mill effluent comprised of following major groups of hydrogen producers: Thermoanaerobacterium spp. (49% and 36%), T. thermosaccharolyticum (16% and 10%), phylum Firmicutes (low G+C) gram positive bacteria (15% and 27%). Extreme-thermophilic (70 °C) hydrogen producing reactors, one fed with xylose and another, fed with lignocellulosic hydrolysate comprised of following major groups of hydrogen producers: Caldicellulosiruptor spp. (40.5% and 20.5%), phylum Firmicutes (low G+C) gram positive bacteria (17% and 20%), Archaea (7% and 8.5%), and Thermoanaerobacterium spp. (0% and 5%). Results obtained, showed good applicability of the probes Tbm1282, Tbmthsacc184 and Ccs432 for specific detection and quantification of thermophilic and extreme-thermophilic hydrogen producers in complex environments.

Start-up of Anaerobic Hydrogen Producing Reactors Seeded with Sewage Sludge

The procedure for starting-up continuously stirred tank reactors (CSTR) for acclimating anaerobic hydrogen-producing microorganisms with sewage sludge was investigated. Initially, feeding with glucose and sucrose as well as mixing were carried out in semicontinuous mode; hydraulic retention time (HRT) was in an order of 20, 15, 10, 5, 2.5 and 2 days. When the pH declined to its lowest value (pH 5.18), it was adjusted to 6.7 using sodium hydroxide (1 N). At the same time, the semi-continuous operation was changed to a continuous one. Finally, the pH was continuously regulated at approximately 6.7. The results indicate that this procedure can be used to cultivate seed sludge for hydrogen production from sewage sludge resulting in a large hydrogen production in less than 60 days. When the substrate was glucose, a hydrogen yield of 1.63 mol H2/mol glucose and a specific hydrogen production rate of 321 mmol H2/g VSS day at an HRT of 13.3 h was achieved. When the substrate was sucrose with the same HTR, a hydrogen yield of 4.45 mol H2/mol sucrose and a specific hydrogen production rate of 707 mmol H2/g VSS day was obtained.