References of "Delvigne, Frank"
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See detailApplication of Steam Explosion as Pretreatment on Lignocellulosic Material: A Review
Jacquet, Nicolas ULg; Maniet, Guillaume ULg; Vanderghem, Caroline ULg et al

in Industrial & Engineering Chemistry Research (in press)

Steam explosion is a thermo-mechanicochemical pretreatment which allows the breakdown of lignocellulosic structural components by the action of heating, formation of organic acids during the process, and ... [more ▼]

Steam explosion is a thermo-mechanicochemical pretreatment which allows the breakdown of lignocellulosic structural components by the action of heating, formation of organic acids during the process, and shearing forces resulting in the expansion of the moisture. Two distinct stages compose the steam-explosion process: vapocracking and explosive decompression which include modification of the material components: hydrolysis of hemicellulosic components (mono- and oligosaccharides released), modification of the chemical structure of lignin, and modification of the cellulose crystallinity index, etc. These effects allow the opening of lignocellulosic structures and influence the enzymatic hydrolysis yield of the material. [less ▲]

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See detailMetabolic variability in bioprocessing : implications of microbial phenotypic heterogeneity
Delvigne, Frank ULg; Zune, Quentin ULg; Lara, Alvaro et al

in Trends in Biotechnology (in press)

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See detailDynamic single-cell analysis of Saccharomyces cerevisiae under process perturbation: comparison of different methods for monitoring the intensity of population heterogeneity
Delvigne, Frank ULg; Baert, Jonathan ULg; Gofflot, Sébastien et al

in Journal of Chemical Technology & Biotechnology (in press)

BACKGROUND: Single cell biology has attracted a lot of attention these past few years and has led to numerous fundamental results pointing out the heterogeneity of clonal cell populations. In this context ... [more ▼]

BACKGROUND: Single cell biology has attracted a lot of attention these past few years and has led to numerous fundamental results pointing out the heterogeneity of clonal cell populations. In this context, microbial phenotypic heterogeneity under bioprocessing conditions needs to be further investigated. In this study, yeast based processes have been investigated by using on-line flow cytometry in combination with a fluorescent transcriptional reporter (GFP) and viability fluorescence tags (propidium iodide, PI). Methods aiming at expressing the dispersion of these fluorescence tags among the yeast populations have been investigated for different bioreactor operating conditions. RESULTS: Yeast viability was determined on the basis of PI uptake. Segregation between PI negative and positive subpopulations could be efficiently quantified on the basis of the mean-to-median ratio or the amplitude of the interquartile range. On the other hand, the same quantification could not be made for the segregation occurring at the level of GFP synthesis. Indeed, when cells were exposed to sub-lethal or mild stresses (such as in scale-down reactors) two GFP subpopulations could be visualized by real-time FC, but quantification by one of the above-mentioned methods was not possible. CONCLUSIONS: Yeast population heterogeneity was observed in representative bioreactor operating conditions. Difficulties for the determination of segregation at the level of GFP synthesis point out the fact that one needs to understand the segregation mechanisms for the applied fluorescent reporters, to judge whether simple mathematical tools may be applied or if more sophisticated computational tools are needed for the quantification of the microbial population segregation. [less ▲]

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See detailFungal biofilm reactor improves the productivity of hydrophobin HFBII
Khalesi, Mohammadreza; Zune, Quentin ULg; Telek, Samuel ULg et al

in Biochemical Engineering Journal (in press)

Production and purification of hydrophobin HFBII has recently been the subject of intensive research, but the yield of production needs to be further improved for a generic use of this molecule at ... [more ▼]

Production and purification of hydrophobin HFBII has recently been the subject of intensive research, but the yield of production needs to be further improved for a generic use of this molecule at industrial scale. In a first step, the influence of different carbon sources on the growth of Trichoderma reesei and the production of HFBII was investigated. The optimum productivity was obtained by using 40 g/L lactose. Carbon starvation and excretion of extracellular enzyme were determined as two main conditions for the production of HFBII. In the second phase, and according to the physiological mechanisms observed during the screening phase, a bioreactor set up has been designed and two modes of cultures have been investigated, i.e. the classical submerged fermentation and a fungal biofilm reactor. In this last set-up, the broth is continuously recirculated on a metal packing exhibiting a high specific surface. In this case, the fungal biomass was mainly attached to the metal packing, leading to a simplification of downstream processing scheme. More importantly, the HFBII concentration increased up to 48.6 ± 6.2 mg/L which was 1.8 times higher in this reactor configuration and faster than the submerged culture. X-ray tomography analysis shows that the biofilm overgrowth occurs when successive cultures are performed on the same packing. However, this phenomenon has no significant influence on the yield of HFBII, suggesting that this process could be operated in continuous mode. Protein hydrolysis during stationary phase was observed by MALDI-TOF analysis according to the removal of the last amino acid from the structure of HFBII after 48 h from the beginning of fermentation in biofilm reactor. Hopefully this modification does not lead to alternation of the main physicochemical properties of HFBII. [less ▲]

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See detailA fungal biofilm reactor based on metal structured packing improves the quality of a Gla::GFP fusion protein produced by Aspergillus oryzae
Zune, Quentin ULg; Delepierre, Anissa ULg; Gofflot, Sebastien et al

in Applied Microbiology and Biotechnology (2015)

Fungal biofilm is known to promote the excretion of secondary metabolites in accordance with solid-state related physiological mechanisms. This work is based on the comparative analysis of classical ... [more ▼]

Fungal biofilm is known to promote the excretion of secondary metabolites in accordance with solid-state related physiological mechanisms. This work is based on the comparative analysis of classical submerged fermentation with a fungal biofilm reactor for the production of a Gla::GFP fusion protein by Aspergillus oryzae. The biofilm reactor comprises a metal structured packing allowing the attachment of the fungal biomass. Since the production of the target protein is under the control of the promoter glaB, specifically induced in solid-state fermentation, the biofilm mode of culture is expected to enhance the global productivity. Although production of the target protein was enhanced by using the biofilm mode of culture, we also found that fusion protein production is also significant when the submerged mode of culture is used. This result is related to high shear stress leading to biomass autolysis and leakage of intracellular fusion protein into the extracellular medium. Moreover, 2D-gel electrophoresis highlights the preservation of fusion protein integrity produced in biofilm conditions. Two fungal biofilm reactor designs were then investigated further, i.e. with full immersion of the packing or with medium recirculation on the packing, and the scale-up potentialities were evaluated. In this context, it has been shown that full immersion of the metal packing in the liquid medium during cultivation allows for a uniform colonization of the packing by the fungal biomass and leads to a better quality of the fusion protein. [less ▲]

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See detailDetermination of Zinc, Cadmium and Lead Bioavailability in Contaminated Soils at the Single-Cell Level by a Combination of Whole-Cell Biosensors and Flow Cytometry
Hurdebise, Quentin ULg; Tarayre, Cédric ULg; Fischer, Christophe ULg et al

in Sensors (2015)

Zinc, lead and cadmium are metallic trace elements (MTEs) that are widespread in the environment and tend to accumulate in soils because of their low mobility and non-degradability. The purpose of this ... [more ▼]

Zinc, lead and cadmium are metallic trace elements (MTEs) that are widespread in the environment and tend to accumulate in soils because of their low mobility and non-degradability. The purpose of this work is to evaluate the applicability of biosensors as tools able to provide data about the bioavailability of such MTEs in contaminated soils. Here, we tested the genetically-engineered strain Escherichia coli pPZntAgfp as a biosensor applicable to the detection of zinc, lead and cadmium by the biosynthesis of green fluorescent protein (GFP) accumulating inside the cells. Flow cytometry was used to investigate the fluorescence induced by the MTEs. A curvilinear response to zinc between 0 and 25 mg/L and another curvilinear response to cadmium between 0 and 1.5 mg/L were highlighted in liquid media, while lead did not produce exploitable results. The response relating to a Zn2+/Cd2+ ratio of 10 was further investigated. In these conditions, E. coli pPZntAgfp responded to cadmium only. Several contaminated soils with a Zn2+/Cd2+ ratio of 10 were analyzed with the biosensor, and the metallic concentrations were also measured by atomic absorption spectroscopy. Our results showed that E. coli pPZntAgfp could be used as a monitoring tool for contaminated soils being processed. [less ▲]

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See detailThermophilic and cellulolytic consortium isolated from composting plants improves anaerobic digestion of cellulosic biomass: toward a microbial resource management approach
Kinet, Romain ULg; Destain, Jacqueline ULg; Hiligsmann, Serge ULg et al

in Bioresource Technology (2015)

A cellulolytic consortium was isolated from a composting plant in order to boost the initial hydrolysis step encountered in anaerobic digestion. Improvement of the cellulose degradation, as well as biogas ... [more ▼]

A cellulolytic consortium was isolated from a composting plant in order to boost the initial hydrolysis step encountered in anaerobic digestion. Improvement of the cellulose degradation, as well as biogas production, was observed for the cultures inoculated with the exogenous consortium. Metagenomics analyses pointed out a weak richness (related to the number of OTUs) of the exogenous consortium induced by the selective pressure (cellulose as sole carbon source) met during the initial isolation steps. Main microbial strains determined were strictly anaerobic and belong to the Clostridia class. During cellulose anaerobic degradation, pH drop induced a strong modification of the microbial population. Despite the fact that richness and evenness were very weak, the exogenous consortium was able to adapt and to maintain the cellulolytic degradation potential. This important result point out the fact that simplified microbial communities could be used in order to increase the robustness of mixed cultures involved in environmental biotechnology. [less ▲]

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See detailInvestigation of the links between mass transfer conditions, dissolved hydrogen concentration and biohydrogen production by the pure strain Clostridium butyricum CWBI1009
Beckers, L.; Masset, J.; Hamilton, C. et al

in Biochemical Engineering Journal (2015), 98

Fermentative hydrogen production has often been described as inhibited by its own gas production. In this work, hydrogen production by Clostridium butyricum was investigated in batch Biochemical Hydrogen ... [more ▼]

Fermentative hydrogen production has often been described as inhibited by its own gas production. In this work, hydrogen production by Clostridium butyricum was investigated in batch Biochemical Hydrogen Potential (BHP) tests and in a 2.5L anaerobic sequenced batch reactor (AnSBR) under different operating conditions regarding liquid-to-gas mass transfer. Through the addition of both stirring up to 400rpm and nitrogen sparging, the yields were enhanced from 1.6 to 3.1molH2molglucose -1 and the maximum hydrogen production rates from 140 to 278mLh-1. These original results were achieved with a pure Clostridium strain. They showed that hydrogen production was improved by a higher liquid-to-gas hydrogen transfer resulting in a lower dissolved hydrogen concentration in the culture medium and therefore in a lower bacterial inhibition. In addition, biohydrogen partitioning between the gas and the liquid phase did not conform to Henry's Law due to critical supersaturation phenomena up to seven-fold higher than the equilibrium conditions. Therefore, dissolved hydrogen concentration should be systematically measured instead of the headspace hydrogen partial pressure. A model was proposed to correlate H2 production yield and rate by the pure C. butyricum strain CWBI1009 with mass transfer coefficient KLa. [less ▲]

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See detailThe use of microorganisms of cassava retting for the production of pectinolytic enzymes
Kouhounde, Sonagnon Hermann Serge ULg; Adeoti, Kifouli; Delvigne, Frank ULg et al

in Journal of Microbiology Biotechnology and Food Sciences (2014), 4(3), 277-281

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See detailLa lutte contre les moustiques (Diptera: Culicidae): diversité des approches et application du contrôle biologique
Bawin, Thomas ULg; Seye, Fawrou; Boukraa, Slimane ULg et al

in Canadian Entomologist (2014)

Many mosquito (Diptera: Culicidae) species are zoonotic vectors responsible for numerous infectious diseases of medical and veterinary importance. Currently, changes in the vectors’ geographical ... [more ▼]

Many mosquito (Diptera: Culicidae) species are zoonotic vectors responsible for numerous infectious diseases of medical and veterinary importance. Currently, changes in the vectors’ geographical distribution induced chiefly by anthropogenic factors are accompanied by emerging and reemerging infectious diseases in Europe and North America. Since the advent of synthetic insecticides during the Second World War, mosquitoes are the object of considerably expanded and deepened research. In an integrated pest management context, means of control are now mainly classified as: (1) environmental management and physical control, (2) chemical control, (3) genetic control, and (4) biological control by means of entomophagous predators and entomopathogenic microorganisms. In this context, these last have significant potential because of their ability to infect and kill their host with more or less targeted selectivity. This article proposes to emphasize biological control among other techniques in mosquito control, and to assess the potential and the opportunities offered by entomopathogenic bacteria, viruses and fungi. Finally, their use as biopesticides is discussed. [less ▲]

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See detailBioRefine Project: Detection of bioavailability of Metallic Trace Elements in soils by the use of microbial biosensors
Tarayre, Cédric ULg; Hurdebise, Quentin ULg; Fischer, Christophe ULg et al

Poster (2014, September 09)

Zinc, lead and cadmium are the main Metallic Trace Elements (MTEs) found in soils contaminated by the mining industry in Europe. MTEs are spread in the environment because of the disruption of ... [more ▼]

Zinc, lead and cadmium are the main Metallic Trace Elements (MTEs) found in soils contaminated by the mining industry in Europe. MTEs are spread in the environment because of the disruption of biogeochemical cycles caused by human activities. Due to their low mobility and biodegradability, they accumulate in soils where they are strongly bound to particles. It has become necessary to understand interactions between MTEs and the environment and to implement remediation actions. This work is focused on remediation monitoring techniques by using whole cell microbial biosensors able to detect zinc, lead and cadmium. Biosensors provide a signal in response to the bio-available concentration in MTEs, which are valuable for the design of efficient techniques involving bioremediation. Whole cell biosensors used in this work are based on Escherichia coli strains carrying a fluorescent reporter system. The reporter element contains a promoter sensitive to MTEs and a gene coding for the Green Fluorescent Protein (GFP). MTEs activate the synthesis of GFP, which is a very stable protein, causing the accumulation of GFP inside the cells. Then, fluorescence can be measured by flow cytometry. In this study, two biosensors were investigated: E. coli pPzraPgfp and E. coli pPzntAgfp. The last strain provided a linear response to zinc up to 20 mg/l and a curvilinear response to cadmium up to 0.15 mg/l. No detection was highlighted regarding lead. In practical cases, soils and wastes are contaminated by several types of MTEs. Consequently, combined contaminations were also tested. This work allowed highlighting that the strain E. coli pPzntAgfp can be used to assess the bioavailability of cadmium in soils, although the experimental procedure must be improved. This work is supported by the BioRefine Project, a European project in which various member states focus on recovery of inorganics from organic wastestreams. We gratefully acknowledge the INTERREG IVB NWE programme, which financed the BioRefine Project (ref. 320J-BIOREFINE). [less ▲]

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See detailDesign of a fungal biofilm reactor for recombinant protein production from Aspergillus oryzae
Zune, Quentin ULg; Delepierre, Anissa ULg; Bauwens, Julien ULg et al

Poster (2014, September 07)

Fungi are microorganisms exhibiting high secretive power of various metabolites and have the ability to perform post-translational modifications during protein synthesis. In the field of fermentation ... [more ▼]

Fungi are microorganisms exhibiting high secretive power of various metabolites and have the ability to perform post-translational modifications during protein synthesis. In the field of fermentation industry, they are ideal hosts for secondary metabolites and recombinant protein production. At the industrial-scale, equipments usually required for solid-state or submerged fermentation of filamentous fungi have demonstrated their limitations in terms of productivity, mass transfers or products recovery (1, 2). Recently, fungal biofilm reactors were designed to combine advantages from submerged and solid-state culture and reveal their usefulness for greater secondary metabolites production relative to submerged culture conditions (3). In our work, we propose the design of a fungal biofilm reactor for a recombinant protein production from an Aspergillus oryzae strain containing a GFP reporter gene system under the control of a promoter specifically induced in solid-state conditions. The fungal biofilm reactor is composed of a metal structured packing, having the function of inert support for biofilm growth, immerged or aspersed by a liquid medium. Whereas recombinant protein production is not significantly different at the flask-scale between submerged and biofilm conditions, productivity is higher in the submerged conditions at the bioreactor-scale. Presence of recombinant proteins entrapped in the biofilm matrix highlights a diffusion constraint and a lower mass transfer in our fungal biofilm reactor. However, persistence of a free liquid biomass of low viscosity and fungal biomass retention on the support are attractive for the implementation of a continuous process in our fungal biofilm reactor. Further studies will consider a 2-D proteomic comparison of the extracellular medium from fungal biofilm reactor and submerged culture conditions in order to better understand proteins secretion and identify over-expressed proteins in biofilm conditions. [less ▲]

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See detailImplications of microbial phenotypic heterogeneity in large-scale bioprocessing conditions
Delvigne, Frank ULg; Gorret, Nathalie; Molina-Jouve, Carole et al

Conference (2014, September)

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See detailMesophilic biohydrogen production by Clostridium butyricum CWBI1009 in trickling biofilter reactor
Puhulwella, Rathnasiri G.; Beckers, Laurent; Delvigne, Frank ULg et al

in International Journal of Hydrogen Energy (2014), 39

This study investigates the mesophilic biohydrogen production from glucose using a strictly anaerobic strain, Clostridium butyricum CWBI1009, immobilized in a trickling bed sequenced batch reactor (TBSBR ... [more ▼]

This study investigates the mesophilic biohydrogen production from glucose using a strictly anaerobic strain, Clostridium butyricum CWBI1009, immobilized in a trickling bed sequenced batch reactor (TBSBR) packed with a Lantec HD Q-PAC® packing material (132 ft2/ft3 specific surface). The reactor was operated for 62 days. The main parameters measured here were hydrogen composition, hydrogen production rate and soluble metabolic products. pH, temperature, recirculation flow rate and inlet glucose concentration at 10 g/L were the controlled parameters. The maximum specific hydrogen production rate and the hydrogen yield found from this study were 146 mmol H2/L.d and 1.67 mol H2/mol glucose. The maximum hydrogen composition was 83%. Following a thermal treatment, the culture was active without adding fresh inoculum in the subsequent feeding and both the hydrogen yield and the hydrogen production rate were improved. For all sequences, the soluble metabolites were dominated by the presence of butyric and acetic acids compared to other volatile fatty acids. The results from the standard biohydrogen production (BHP) test which was conducted using samples from TBSBR as inoculum confirmed that the culture generated more biogas and hydrogen compared to the pure strain of C. butyricum CWBI1009. The effect of biofilm activity was studied by completely removing (100%) the mixed liquid and by adding fresh medium with glucose. For three subsequent sequences, similar results were recorded as in the previous sequences with 40% removal of spent medium. The TBSBR biofilm density varied from top to bottom in the packing bed and the highest biofilm density was found at the bottom plates. Moreover, no clogging was evidenced in this packing material, which is characterized by a relatively high specific surface area. Following a PCA test, contaminants of the Bacillus genus were isolated and a standard BHP test was conducted, resulting in no hydrogen production. [less ▲]

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See detailPathogenicity of Aspergillus clavatus produced in a fungal biofilm bioreactor toward Culex quinquefasciatus (Diptera: Culicidae)
Seye, Fawrou; Bawin, Thomas ULg; Boukraa, Slimane ULg et al

in Journal of Pesticide Science (2014), 39(3), 127-132

Many entomopathogenic fungi have been demonstrated to be potential agents for efficiently controlling mosquito populations. In the present study, we investigated a bioreactor system to produce metabolites ... [more ▼]

Many entomopathogenic fungi have been demonstrated to be potential agents for efficiently controlling mosquito populations. In the present study, we investigated a bioreactor system to produce metabolites and conidia by combining technological advantages of submerged and solid-state fermentations. The efficiency of fungal products was tested toward mosquitoes. Aspergillus clavatus (Eurotiales: Trichocomaceae) was grown by semi-solid-state fermentation in a bioreactor for up to 7 days. Depending on conidial doses (2.5×10^7, 5×10^7, 7.5×10^7, 10×10^7 and 12.5×10^7 conidia/mL), mortality ranged from 37.2±15.0 to 86.3±5.0% toward larvae and from 35.8±2.0 to 85.2±1.5% toward adults. The metabolites (10, 20, 40, 60, 80 and 100% v/v) yielded mortality from 23.7±15.0 to 100.0±0.1% toward larvae, and two sprayed volumes (5 and 10 mL) reached 45.5±1.4 and 75.6±2.6% mortality, respectively, toward adults. [less ▲]

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See detailDevelopment of entomopathogenic fungi in mosquito control: which kind of production for which efficiency?
Bawin, Thomas ULg; Delvigne, Frank ULg; Francis, Frédéric ULg

Poster (2014, August 06)

Mosquitoes (Diptera: Culicidae) are zoonotic vectors responsible for numerous infectious diseases of medical and veterinary importance such as filariasis, malaria and encephalitis. As part of an ... [more ▼]

Mosquitoes (Diptera: Culicidae) are zoonotic vectors responsible for numerous infectious diseases of medical and veterinary importance such as filariasis, malaria and encephalitis. As part of an integrated vector control, entomopathogenic fungi could be developed as biopesticides in two ways: spores and metabolites recognized as effective virulence factors. Solid-state fermentation enhances spore production and induces the secretion of metabolites quantitatively and qualitatively different from submerged fermentation, which impairs fungal metabolic efficiency. In this context, we showed high spore productivity of solid-state media based on agro-industrial substrates as wheat bran. Spores remained pathogenic, as revealed by classical toxicity tests and electron microscopy. However, the absence of free water makes culture parameter variations difficult to control in large-scale. Recently, we performed a bioreactor design intended for simultaneous spore and metabolite production, combining the technological advantages of submerged and solid-state fermentations. Biofilm fermentation (i.e. growth of fungal biomass on an inert support immerged in a nutrient medium) is a tremendous production system favouring the secretion of insecticidal metabolites in the liquid medium as we showed recently. This is also an interesting tool to provide an overview of the complexity of the metabolic pathways involved in the regulation of extracellular metabolites secretion because corresponding genes are reported to be differentially expressed from classical fermentation systems. Researches in vector control are currently intensified. In this context, the identification of genes and metabolites specifically expressed during biofilm fermentation will help to develop new technologies related both to the design of bioreactor and the production of insecticidal proteins. [less ▲]

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