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See detailLa production d'hydrogène par fermentation anaérobie: Voies d'optimalisation et d'application du bioprocédé
Beckers, Laurent ULg

Doctoral thesis (2013)

The emergence of environmental and societal issues caused by the fossil fuels consumption and the simultaneous increase of the energetic needs will lead the society to evolve into a new energetic system ... [more ▼]

The emergence of environmental and societal issues caused by the fossil fuels consumption and the simultaneous increase of the energetic needs will lead the society to evolve into a new energetic system. The creation of the hydrogen society could bring a suitable and sustainable solution since the production and use of hydrogen could be operated at higher yields than the fossil fuels economy and produce energy while generating only water vapour as co product. However, in order to get rid of the fossil fuels consumption, there is a need to diversify the hydrogen production processes and technology, currently still based on CO2—emitting technologies. The so-called “dark fermentation” process is based on strict of facultative anaerobic bacteria producing biohydrogen and soluble metabolites as a fermentation co product. These microorganisms consume organic substrates such as in wastewater to achieve their growth. The biohydrogen technology has been studied during several years in laboratory but still is not mature to be brought at an industrial scale. Indeed, there is first a need to improve the performances (such as the H2 yields and production rates) to achieve the technical and economical requirements. This thesis investigates and discusses various possibilities in order to bring the biohydrogen production process to a larger scale. The strain investigated in this work, Clostridium butyricum, can achieve high performances (in terms of hydrogen yields, about 1.9 to 2.2 molH2·molglucose -1 and production rates, about 50 to 110 mLH2·L-1·h-1 in batch or sequencing-batch cultures) and is able to consume simple and more complex substrates. However, being a strict anaerobic strain, its uses in pure culture requires the achievement of strong anaerobic conditions using artificial and costly means. Moreover, even if the work in pure culture has some advantages at the laboratory scale, it is inappropriate to larger volumes of bioreactor. Therefore, mixed cultures were investigated in batch and sequencing-batch bioreactors, resulting in a decrease by about 30 to 50% of the yields (down to 1.2 to 1.7 molH2·molglucose -1). The mixed cultures reached however comparable or higher performances than the scientific literature confirming the interest of the approach suggested in this work. However, the performances need to be further enhanced in order to make the process economically possible. Therefore, improvements of the yields and the rates were proposed. On the one hand, the yields were increased by 55 to 100% (up to 3.1 molH2·molglucose -1) by improving the mass transfer conditions and, by the way, decreasing the dissolved hydrogen concentration in the liquid media. These considerations led to the design of a novel biodisc bioreactor working continuously and allowing the efficient hydrogen mass transfer. In mixed culture, the biodisc bioreactor reached high performances (H2 yields of 2.4 molH2·molglucose -1 and H2 rates of 600 mLH2·L-1·h-1), showing the interest of the original design and of the “mass transfer” approach achieved in this work. On the other hand, the addition in the culture medium of small quantities of metallic nanoparticles showed a catalytic-like effect by enhancing the hydrogen production rate by about 40 to 100%. Ending this work, the general discussion evidences the advantages of the different techniques suggested in the work and compares them to the recent scientific literature. Furthermore, perspectives are given in terms of scientific outlooks, considering the economical, environmental and technical aspects, in order to bring the hydrogen production process at a large scale. [less ▲]

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See detailInvestigating the factors for fermentative biohydrogen improvement: original bioreactors design and hydrogen partial pressure effect
Beckers, Laurent ULg; Hiligsmann, Serge ULg; Hamilton, Christopher ULg et al

in WHEC 2012, Toronto June 3rd - 7th (2012, June 05)

The anaerobic production of hydrogen from biomass offers the potential production of usable biogas from a variety of renewable resources. However, in order to produce hydrogen at high yields and ... [more ▼]

The anaerobic production of hydrogen from biomass offers the potential production of usable biogas from a variety of renewable resources. However, in order to produce hydrogen at high yields and production rates the biotechnological process needs to be further optimized and efficient bioreactors must be designed [1]. At the CWBI, a continuous horizontal rotating cylinder bioreactor has been designed and investigated to produce biohydrogen from glucose by the strain Clostridium butyricum [2] at good yields (1,9molH2•molglucose-1) and production rates (48,6mmolH2•Lmilieu-1.molhexose-1•h-1). This reactor has an internal volume of 2.3L and a small working volume (300ml) (fig.1). It enhances the hydrogen production rates (by about three times more than a completely stirred bioreactor) by partially immobilizing the bacteria on the porous support. Moreover, the rotating cylinder design enables efficient H2 gas transfer from the liquid phase increasing hydrogen yields by about 25% compared to a completely stirred bioreactor [3-4]. Other original bioreactors, such as a trickle bed, have been built with the same aim of lowering the hydrogen partial pressure and led to similar results. Our work shows the importance of a good liquid to gas transfers in the biohydrogen-producing reactors to reach higher performances. [less ▲]

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See detailThe MicroH2 project:an association of four laboratories to improve theknowledge on biohydrogen production precesses
Beckers, Laurent ULg; Calusinska, Magdalena ULg; Hamilton, Christopher ULg et al

Poster (2012, June 04)

This poster presents a collaborative research project (MicroH2) held at the University of Liège (Belgium) since 2007 (www.microh2.ulg.ac.be) and involving four different research groups. The project aims ... [more ▼]

This poster presents a collaborative research project (MicroH2) held at the University of Liège (Belgium) since 2007 (www.microh2.ulg.ac.be) and involving four different research groups. The project aims to develop a center of excellence in the fields of photo- and dark- biohydrogen production. Our studies contribute to improve the knowledge of the processes involved in the microbiological production of hydrogen, from a fundamental and practical point of view. Some results are highlighted here. The research concerning photofermentation focuses on the interactions between respiration, photosynthesis and H2-producing pathways in algal microorganisms, by using mitochondrial mutants and genetically modified strains with modified ability for hydrogen production [1-2]. To study the metabolism of the hydrogen production by anaerobic bacteria, pure cultures and defined consortia are used and their production of biogas and soluble metabolites is measured. Moreover, we have developed and optimized molecular tools, like quantitative RT-PCR and FISH, to monitor the variations of bacterial populations in novel bioreactors for hydrogen production [3-4]. We have also mined the complete genomes of Clostridium spp. for putative hydrogenase genes and found a large diversity of them [5]. [less ▲]

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See detailEffects of hydrogen partial pressure on fermentative biohydrogen production by a chemotropic Clostridium bacterium in a new horizontal rotating cylinder reactor
Beckers, Laurent ULg; Hiligsmann, Serge ULg; MAsset, Julien et al

in Energy Procedia (2012)

In order to produce fermentative biohydrogen at high yields and production rates, efficient bioreactors have to be designed. A new reactor called anaerobic biodisc reactor allowed the production of ... [more ▼]

In order to produce fermentative biohydrogen at high yields and production rates, efficient bioreactors have to be designed. A new reactor called anaerobic biodisc reactor allowed the production of biohydrogen from glucose with the selected Clostridium sp. strain at high yields (2.49 molH2•molglucose-1) and production rates (598 mlH2•medium-1•h-1). The bacteria were fixed on a rotating support enabling efficient gas transfer from the liquid to the phase. It allowed the metabolism of the bacteria to produce more hydrogen. Moreover, an increase of the total pressure 0.18 bar lowered the yields of 19.5% while a decrease of 0.11 bar increased the yields of 7%. Our work concludes on the importance of providing good liquid to gas transfers in the biohydrogen-producing reactors. [less ▲]

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See detailLa production de biohydrogène à partir de substrats carbohydratés : état de l'art
Hiligsmann, Serge ULg; Beckers, Laurent ULg; Masset, Julien ULg et al

in Récents progrès en génie des procédés (2011, December 01)

Hydrogen is being considered as an ideal and clean energy carrier. The recent advances to produce biohydrogen from renewable sources such as biomass and particularly by fermentation of carbohydrate-rich ... [more ▼]

Hydrogen is being considered as an ideal and clean energy carrier. The recent advances to produce biohydrogen from renewable sources such as biomass and particularly by fermentation of carbohydrate-rich substrates from agriculture and agro-industries appear promising. Such a process enables both organic waste treatment and renewable energy production to be coupled. The paper presents the state of the art about the different hydrogen-producing microorganisms and the parameters that have been investigated in order to improve the hydrogen production yields and rates. [less ▲]

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See detailEffets de la pression partielle en hydrogène sur la production anaérobie de bioH2 par une bactérie chimiotrophe du genre Clostridium dans un nouveau réacteur à cylindre horizontal rotatif.
Beckers, Laurent ULg; Hiligsmann, Serge ULg; Hamilton, Christopher ULg et al

in Récents Progrès en Génie des Procédés (2011, December), 101

Hydrogen is widely considered as the most promising energetic carrier. At an industrial scale, steam reforming of methane is currently the major hydrogen producing process. But it may also be produced ... [more ▼]

Hydrogen is widely considered as the most promising energetic carrier. At an industrial scale, steam reforming of methane is currently the major hydrogen producing process. But it may also be produced from renewable biomass. Indeed, the fermentative production of hydrogen from renewable biomass using anaerobic bacteria could at least partially reduce our dependence on fossil fuel, decrease the carbon dioxide emissions and produce “green” energy. It offers the potential production of usable hydrogen from a variety of renewable resources such as carbohydrates wastes from agriculture or agro-food industries. This technology is based on anaerobic fermentation, called dark fermentation, by chemotrophic bacteria. The investigations carried out at CWBI involve selection and characterization of bacteria strains able to produce biohydrogen efficiently and with a wide range of substrate. The selected strains at the laboratory has been characterised as Clostridium sp. In order to produce hydrogen at high yields and production rates, the biotechnological process needs to be further optimized and efficient bioreactors must be designed. At CWBI, a new reactor called “horizontal rotating cylinder bioreactor” allows the production of biohydrogen from glucose with our Clostridium sp. strain with a high yield and production rate. This reactor, working continuously, has an internal volume of 2.3l but a working volume (liquid phase) of 300ml. Firstly, it enhances the hydrogen production rate by partially fixing the bacteria on the porous cylinder and thus increasing the cell concentration in the bioreactor. Secondly, the rotative cylinder enables efficient gas transfer (mainly hydrogen) from the liquid phase where it is produced by the bacteria. This is an important way to enhance hydrogen production yield by allowing the bacteria metabolism to shift in a fermentation type that produces more hydrogen. This was confirmed by increasing or decreasing the total pressure in the bioreactor and observing the influence of hydrogen production. The liquid to gas hydrogen transfer is possibly an important factor to enhance the biogas production. Our investigation confirmed this by testing different liquid to gas transfer condition in BHP test (batch fermentation in 250ml serum bottles).This was made either by decreasing total and partial pressure or by increasing the mixing state of the media. Our work concludes the importance of providing good liquid to gas transfers in the biohydrogen producing reactors to enhance the hydrogen production and reach higher yields and production rates. [less ▲]

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See detailEffects of hydrogen partial pressure on fermentative biohydrogen production by a chemotropic Clostridium bacterium in a new horizontal rotating cylinder reactor
Beckers, Laurent ULg; Hiligsmann, Serge ULg; Hamilton, Christopher ULg et al

Poster (2011, December)

The fermentative production of hydrogen using chemotrophic anaerobic bacteria offers a new way to produce “green” energy from a large variety of renewable resources and organic wastes. In order to produce ... [more ▼]

The fermentative production of hydrogen using chemotrophic anaerobic bacteria offers a new way to produce “green” energy from a large variety of renewable resources and organic wastes. In order to produce hydrogen at high yields and production rates, efficient bioreactors must be designed. A new reactor called “horizontal rotating cylinder bioreactor” allows the production of biohydrogen from glucose with the selected Clostridium sp. strain at high yields (1,9molH2·molglucose-1) and production rates (48,6mmolH2·lmilieu-1.molhexose-1·h-1). The rotative cylinder where the bacteria are fixed enables efficient gas transfer (mainly hydrogen) from the liquid phase where it is produced by the bacteria. This is an important way to allow the bacteria metabolism to shift in a fermentation pathway that produces more hydrogen. This was confirmed by varying the total pressure in the bioreactor. An increase of the total pressure 0,18bar lowered the yields of 19,5% while a decrease of 0,11bar increased the yields of 7%. Our work concludes the importance of providing good liquid to gas transfers in the biohydrogen-producing reactors in order to reach higher yields and production rates. [less ▲]

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See detailFermentative biohydrogen production in a novel biodisc bioreactor: Principle and Improvement
Beckers, Laurent ULg; Hiligsmann, Serge ULg; Masset, Julien ULg et al

in Bozhou, Li (Ed.) Low Carbon Earth Summit 2011 Proceeding (2011, October 23)

In order to produce hydrogen at high yields and production rates, the biotechnological process needs to be further optimized and efficient bioreactors must be designed. A biodisc bioreactor has been ... [more ▼]

In order to produce hydrogen at high yields and production rates, the biotechnological process needs to be further optimized and efficient bioreactors must be designed. A biodisc bioreactor has been design and investigated to produce biohydrogen from glucose by the Clostridium butyricum CWBI1009 strain at a high yield and production rate. This reactor, working continuously, has an internal volume of 2.3l but a working volume (liquid phase) of 300ml. Firstly, it enhances the hydrogen production rate (by about 3 times more than a completely stirred bioreactor) by partially fixing the bacteria on the porous support and thus increasing the cell concentration in the bioreactor (decoupling of HRT and SRT). Secondly, the rotating biodisc design enables efficient gas transfer (hydrogen and carbon dioxyde) from the liquid phase where it is produced by the bacteria to the headspace. Indeed, this is an important way to increase hydrogen production yields (by about 25% compared to a completely stirred bioreactor) by allowing the bacteria to focus on the metabolites pathways that produce more hydrogen. Other reactors designs have shown such good results by increasing the interfacial surface. [less ▲]

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See detailComparative study of biological hydrogen production by pure strains and consortia of facultative and strict anaerobic bacteria
Hiligsmann, Serge ULg; Masset, Julien ULg; Hamilton, Christopher ULg et al

in Bioresource Technology (2011), 102

In this paper, a simple and rapid method was developed in order to assess in comparative tests the production of binary biogas mixtures containing CO2 and another gaseous compound such as hydrogen or ... [more ▼]

In this paper, a simple and rapid method was developed in order to assess in comparative tests the production of binary biogas mixtures containing CO2 and another gaseous compound such as hydrogen or methane. This method was validated and experimented for the characterisation of the biochemical hydrogen potential of different pure strains and mixed cultures of hydrogen-producing bacteria (HPB) growing on glucose. The experimental results compared the hydrogen production yield of 19 different pure strains and sludges : facultative and strict anaerobic HPB strains along with anaerobic digester sludges thermally pre-treated or not. Significant yields variations were recorded even between different strains of the same species by i.e. about 20% for three Clostridium butyricum strains. The pure Clostridium butyricum and pasteurianum strains achieved the highest yields i.e. up to 1,36 mol H2/mol glucose compared to the yields achieved by the sludges and the tested Escherichia and Citrobacter strains. [less ▲]

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See detailInvestigation of anaerobic digestion in a two-stage bioprocess producing hydrogen and methane
Hiligsmann, Serge ULg; Hamilton, Christopher ULg; Beckers, Laurent ULg et al

Conference (2010, November 17)

Hydrogen has received wide attention in the last decades as a clean energy vector. The major advantage of energy generation from hydrogen is the near-zero carbon emissions, since the utilization of ... [more ▼]

Hydrogen has received wide attention in the last decades as a clean energy vector. The major advantage of energy generation from hydrogen is the near-zero carbon emissions, since the utilization of hydrogen, either via combustion or via fuel cells, results in pure water. Recently, there has been increasing interest on the biological production of hydrogen gas from renewable biomass such as carbohydrates from agriculture or agro-food industries. This specific anaerobic digestion is called dark fermentation and is involved in the classic anaerobic digestion producing methane. Indeed, in a two-step process, i.e. when acido- and aceto-genesis are carried out in a different bioreactor than methanogenesis, it is feasible to generate separated biogas flows containing either H2 or CH4 depending on specific operating parameters. This paper deals with the comparison of the first stage performances whether operated in optimum conditions for H2 production with a mixed culture or a pure Clostridium butyricum strain. Hydrogen yields of about 1.75 and 2.3 mol H2/mol glucose were achieved respectively. The metabolites, mainly acetate and butyrate, contained in the spent medium were efficiently converted to methane in a second anaerobic digester with a methane yield of about 170 ml/g COD initially fed in the first stage. [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 detailFermentative hydrogen production by Clostridium butyricum CWBI1009 and Citrobacter freundii CWBI952 in pure and mixed cultures
Beckers, Laurent ULg; Hiligsmann, Serge ULg; Hamilton, Christopher ULg et al

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (2010), 14(S2), 541-548

This paper investigates the biohydrogen production by two mesophilic strains, a strict anaerobe (Clostridium butyricum CWBI1009) and a facultative anaerobe (Citrobacter freundii CWBI952). They were ... [more ▼]

This paper investigates the biohydrogen production by two mesophilic strains, a strict anaerobe (Clostridium butyricum CWBI1009) and a facultative anaerobe (Citrobacter freundii CWBI952). They were cultured in pure and mixed cultures in serum bottles with five different carbon sources. The hydrogen yields of pure C. freundii cultures ranged from 0.09 molH2.molhexose-1 (with sucrose) to 0.24 molH2.molhexose-1 (with glucose). Higher yields were obtained by the pure cultures of Cl. butyricum ranging from 0.44 molH2.molhexose-1 (with sucrose) to 0.69 molH2.molhexose-1 (with lactose). This strain also fermented starch whereas C. freundii did not. However, it consumed the other substrates faster and produced hydrogen earlier than Cl. butyricum. This ability has been used to promote the growth conditions of Cl. butyricum in co-culture with C. freundii, since Cl. butyricum is extremely sensitive to the presence of oxygen which strongly inhibits H2 production. This approach could avoid the addition of any expensive reducing agents in the culture media such as L-cysteine since C. freundii consumes the residual oxygen. Thereafter, co-cultures with glucose and starch were investigated: hydrogen yields decreased from 0.53 molH2.molhexose-1 for pure Cl. butyricum cultures to 0.38 molH2.molhexose -1 for mixed culture with glucose but slightly increased with starch (respectively 0.69 and 0.73 molH2.molhexose-1). After 48 h of fermentation, metabolites analysis confirmed with microbial observation, revealed that the cell concentration of C. freundii dramatically decreased or was strongly inhibited by the development of Cl. butyricum. [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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