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See detailImprovement of fermentative biohydrogen production by Clostridium butyricum CWBI1009 in sequenced-batch, horizontal fixed bed and biodisc-like anaerobic reactors with biomass retention
Hiligsmann, Serge ULg; Beckers, Laurent; Masset, Julien et al

in International Journal of Hydrogen Energy (2014), 39

A horizontal tubular fixed bed bioreactor (HFBR) and an anaerobic biodisc-like reactor (AnBDR) were designed to both fix Clostridium biomass and enable rapid transfer of the hydrogen produced to gas phase ... [more ▼]

A horizontal tubular fixed bed bioreactor (HFBR) and an anaerobic biodisc-like reactor (AnBDR) were designed to both fix Clostridium biomass and enable rapid transfer of the hydrogen produced to gas phase in order to decrease the strong effect of H2 partial pressure and H2 supersaturation on the performances of Clostridium strains. The highest H2 production rate (703 mL H2/L.h) and yield (302 mL/g glucose consumed i.e. 2.4 mol/mol) with the pure culture were recorded in the AnBDR with 300 mL culture medium (total volume 2.3 L) at pH 5.2 and a glucose loading rate of 2.87 g/L.h. These results are about 2.3 and 1.3-fold higher than those achieved in the same bioreactor with 500 mL liquid medium and with the same glucose consumption rate. Therefore, our experimentations and a short review of the literature reported in this paper emphasize the relevance of performing bioreactors with high L/G transfer. [less ▲]

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See detailA novel screening method for hydrogenase-deficient mutants in Chlamydomonas reinhardtii based on in vivo chlorophyll fluorescence and photosystem II quantum yield
Godaux, Damien ULg; Emonds-Alt, Barbara ULg; Berne, Nicolas ULg et al

in International Journal of Hydrogen Energy (2013), 38

In Chlamydomonas reinhardtii, prolonged anaerobiosis leads to the expression of enzymes <br />Received 30 August 2012 belonging to various fermentative pathways. Among them, oxygen-sensitive hydrogenases ... [more ▼]

In Chlamydomonas reinhardtii, prolonged anaerobiosis leads to the expression of enzymes <br />Received 30 August 2012 belonging to various fermentative pathways. Among them, oxygen-sensitive hydrogenases <br />Received in revised form (HydA1/2) catalyze the synthesis of molecular hydrogen from protons and reduced ferre- <br />12 November 2012 doxin in the stroma. In this work, by analyzing wild type and mutants affected in H2 <br />Accepted 16 November 2012 production, we show that maximal PSII photosynthetic electron transfer during the first <br />Available online 21 December 2012 seconds of illumination after a prolonged dark-anaerobiosis period is linearly related to <br />hydrogenase capacity. Based on the specific chlorophyll fluorescence induction kinetics <br />Keywords: typical of hydrogenase-deficient mutants, we set up an in vivo fluorescence imaging <br />Chlamydomonas reinhardtii screening protocol allowing to isolate mutants impaired in hydrogenase expression or <br />Anaerobic photosynthesis activity, as well as mutants altered in related metabolic pathways required for energy <br />Hydrogenase production in anaerobiosis. Compared to previously described screens for mutants <br />Chlorophyll fluorescence impaired in H2 production, our screening method is remarkably fast, sensitive and non- <br />Microalgae invasive. Out of 3000 clones from a small-sized insertional mutant library, five mutants <br />Hydrogen photoproduction were isolated and the most affected one was analyzed and shown to be defective for the <br />hydrogenase HydG assembly factor. [less ▲]

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See detailInfluence of the carbon texture of platinum/carbon aerogel electrocatalysts on their behaviour in a Proton Exchange Membrane Fuel Cell cathode
Ouattara-Brigaudet, M.; Berthon-Fabry, S.; Beauger, C. et al

in International Journal of Hydrogen Energy (2012), 37

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See detailNovel FISH and quantitative PCR protocols to monitor artificial consortia composed of different hydrogen-producing Clostridium spp.
Savichtcheva, Olga; Joris, Bernard ULg; Wilmotte, Annick ULg et al

in International Journal of Hydrogen Energy (2011), 36

The use of an artificial consortium composed of selected hydrogen-producing species, instead of a natural anaerobic sludge, has been proposed for biohydrogen production. In order to monitor such a ... [more ▼]

The use of an artificial consortium composed of selected hydrogen-producing species, instead of a natural anaerobic sludge, has been proposed for biohydrogen production. In order to monitor such a consortium composed of different Clostridium spp., new protocols were tested for two different assays, FISH and qPCR. New species-specific FISH probes and qPCR primer sets were developed and optimised for three strains: Clostridium butyricum, Clostridium felsineum and Clostridium pasteurianum, that were used in a consortium. Application of a fast two-step FISH protocol, with pre-treatment step at 90 C for 5 min and a subsequent hybridisation step at higher temperature (55 C) for 20 min resulted in a much shorter analytical time compared to the standard FISH procedure (46 C for 2e3 h) and gave a high hybridisation performance. Moreover, to accurately quantify each microorganism by qPCR assay, two innovations were tested: the direct use of cell lysates (omitting the DNA extraction step) and the use of two alternative molecular markers, recA and gyrA. These markers are present in single copies in the genome, whereas there are multiple copies of the ribosomal operons. This resulted in the development of accurate, reliable and fast FISH and qPCR assays for routine monitoring of the dynamics of artificial hydrogen-producing microbial consortia. Moreover, both techniques can be easily adapted to new Clostridium strains. [less ▲]

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See detailFunctional analysis of hydrogen photoproduction in respiratory-deficient mutants of Chlamydomonas reinhardtii
Lecler, Renaud ULg; Godaux, Damien ULg; Vigeolas, Hélène ULg et al

in International Journal of Hydrogen Energy (2011), 36

In this paper, mitochondrial mutants of Chlamydomonas reinhardtii defective for respiratory complex I (NADH:ubiquinone oxidoreductase), complex III (ubiquinol cytochrome c oxidoreductase) and both ... [more ▼]

In this paper, mitochondrial mutants of Chlamydomonas reinhardtii defective for respiratory complex I (NADH:ubiquinone oxidoreductase), complex III (ubiquinol cytochrome c oxidoreductase) and both complexes I and III were analyzed for H2 photoproduction. Several parameters were followed during the S-deficiency stage and the anaerobic stage leading to H2 photoproduction. At the early aerobic S-deficiency stage, starch and neutral lipids accumulated in all strains but their amount was significantly decreased in mutants compared to wild type. During the H2 photoproduction process, whereas starch content strongly decreased in all strains, neutral lipid amount remained nearly unchanged, suggesting that starch degraded by glycolysis is the preferential substrate for energy production during anaerobiosis. The mutants displayed a decrease in H2 photoproduction correlating to the number of active mitochondrial proton-pumping sites lost in the strains. Our results thus highlight the critical role of oxidative phosphorylation during the first (aerobic) stage of S-starvation when carbon resources are accumulated. [less ▲]

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See detailEffect of pH on glucose and starch fermentation in batch and sequenced-batch mode with a recently isolated strain of hydrogen-producing Clostridium butyricum CWBI1009
Masset, Julien ULg; Hiligsmann, Serge ULg; Hamilton, Christopher ULg et al

in International Journal of Hydrogen Energy (2010), 35(8), 3371-3378

This paper reports investigations carried out to determine the optimum culture conditions for the production of hydrogen with a recently isolated strain Clostridium butyricum CWBI1009. The production ... [more ▼]

This paper reports investigations carried out to determine the optimum culture conditions for the production of hydrogen with a recently isolated strain Clostridium butyricum CWBI1009. The production rates and yields were investigated at 30 °C in a 2.3 l bioreactor operated in batch and sequenced-batch mode using glucose and starch as substrates. In order to study the precise effect of a stable pH on hydrogen production, and the metabolite pathway involved, cultures were conducted with pH controlled at different levels ranging from 4.7 to 7.3 (maximum range of 0.15 pH unit around the pH level). For glucose the maximum yield (1.7 mol H2 mol-1 glucose) was measured when the pH was maintained at 5.2. The acetate and butyrate yields were 0.35 mol acetate mol-1 glucose and 0.6 mol butyrate mol-1 glucose. For starch a maximum yield of 2.0 mol H2 mol-1 hexose, and a maximum production rate of 15 mol H2 mol-1 hexose h-1 were obtained at pH 5.6 when the acetate and butyrate yields were 0.47 mol acetate mol-1 hexose and 0.67 mol butyrate mol-1 hexose. [less ▲]

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See detailOptimisation of culture conditions for biological hydrogen production by Citrobacter freundii CWBI952 in batch, sequenced-batch and semicontinuous operating mode
Hamilton, Christopher ULg; Hiligsmann, Serge ULg; Beckers, Laurent ULg et al

in International Journal of Hydrogen Energy (2010), 35

Investigations were carried out to determine the effect of the pH, the nitrogen source, iron and the dilution rate (h 1) on fermentative hydrogen production from glucose by the newly isolated strain ... [more ▼]

Investigations were carried out to determine the effect of the pH, the nitrogen source, iron and the dilution rate (h 1) on fermentative hydrogen production from glucose by the newly isolated strain Citrobacter freundii CWBI952. The hydrogen production rate (HPR), hydrogen yield, biomass and soluble metabolites were monitored at 30 C in 100 mL serum bottles and in a 2.3 L bioreactor operated in batch, sequenced-batch and semicontinuous mode. The results indicate that hydrogen production activity, formate biosynthesis and glucose intake rates are very sensitive to the culture pH, and that additional formate bioconversion and production of hydrogen with lower biomass yields can be obtained at pH 5.9. In a further series of cultures casein peptone was replaced by (NH4)2SO4, a low cost alternative nitrogen source. The ammonia-based substitute was found to be suitable for H2 production when a concentration of 0.045 g/L FeSO4 was provided. Optimal overall performances (ca. an HPR of 33.2 mL H2/L h and a yield of 0:83 molH2 =molglucose) were obtained in the semicontinuous culture applying the previously optimized parameters for pH, nitrogen, and iron with a dilution rate of 0.012 h 1 and degassing of biogas by N2 at a 28 mL/min flow rate. [less ▲]

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