References of "Thonart, Philippe"
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See detailBiofilm formation on metal structured packing for the production of high added value biomolecules
Zune, Quentin ULg; Zune, Quentin ULg; Toye, Dominique ULg et al

in Récents Progrès en Génie des Procédés (2013)

Many white biotechnology bioprocesses apply techniques from chemical engineering based on bioreactors with mechanical stirring system commonly employed in pharmaceutical sector, food industry or energy ... [more ▼]

Many white biotechnology bioprocesses apply techniques from chemical engineering based on bioreactors with mechanical stirring system commonly employed in pharmaceutical sector, food industry or energy field (Dasilva, 2004). As in chemical engineering, scale-up of these bioprocesses induces physicochemical constraints that affect physiological pathways and decrease performances. In this context, it is essential to think new bioprocesses better suited to physiology of microorganisms, minimizing physicochemical constraints. The aim of this work consists to use stainless steel structured packing (SSP) with high specific area (500-750 m²/m³) as inert support for biomass immobilization in order to produce high added value biomolecules. These bioreactors are biocatalysts in which microbial system is immobilized biomass on the form of a biofilm performing bioconversion of a substrate into a specific product (Rosche, 2009). In this study, an experimental setting containing a SSP reproduces solid-state fermentation (SSF) like conditions. Two well known microorganisms for their ability to form biofilm and secrete metabolites are tested in the experimental setting : Bacillus subtilis for its lipopeptides and Aspergillus oryzae for its glucoamylase. Effectiveness of the bioprocess in term of dynamic of the excretion of the target biomolecule is compared with a classical submerged culture (SmF). For lipopeptides production from B. subtilis, SSP is located in a 20L bioreactor continuously aspersed by liquid medium required to the growth of the biofilm. In the case of A. oryzae, the SSP is partially immerged in a 250 mL shake flask. X-ray tomography of the SSP allows non-invasive visualization and quantification of biofilm repartition inside the support. Implementation of SSP permits almost total immobilization of biomass on the form of a mono-species biofilm to the detriment of the liquid phase. Processing of images obtained by X-ray tomography of the SSP provides relevant information for the optimization of the bioprocess. For both microorganism species, results indicate the influence of parameters such as hydrodynamics, aeration rate and microorganism specificity, on the biofilm morphology inside the support and the performances of the bioprocess. SSF-like conditions in the experimental setting lead to technologic progress, such as absence of foam formation, persistence of the microbial system, and improve the dynamic of metabolites excretion compared with conditions imposed by the submerged culture. Further experiment will consider hydrodynamics aspects and amount of carbon source on effectiveness of the bioprocess. [less ▲]

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See detailSymbiont Diversity in Reticulitermes santonensis (Isoptera: Rhinotermitidae): Investigation Strategy Through Proteomics.
Bauwens, Julien ULg; Millet, Catherine ULg; Tarayre, Cédric ULg et al

in Environmental entomology (2013), 42(5), 882-7

The complex microbial community living in the hindgut of lower termites includes prokaryotes, flagellates, yeasts, and filamentous fungi. Many microorganisms are found in the termite gut, but only a few ... [more ▼]

The complex microbial community living in the hindgut of lower termites includes prokaryotes, flagellates, yeasts, and filamentous fungi. Many microorganisms are found in the termite gut, but only a few are thought to be involved in symbiotic association to participate in cellulose digestion. Proteomics provides analyses from both taxonomical and functional perspectives. We aimed to identify symbiont diversity in the gut of Reticulitermes santonensis (Feytaud), via complementary electrospray ionization associated to ion trap tandem mass spectrometry (LC-MS/MS) and two-dimensional gel electrophoresis associated to matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry analysis. One specific challenge to the study of lower termites is the relatively few data available on abundant symbiotic flagellates. Analysis based on LC-MS/MS revealed few protein families showing assignments to eukaryotes and the taxonomic origin of highly represented actins could not be established. Tubulins proved to be the most suitable protein family with which to identify flagellate populations from hindgut samples using LC-MS/MS, compared with other protein families, although this method targeted few prokaryotes in our assay. Similarly, two-dimensional gel electrophoresis associated to matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry did not succeed in identifying flagellate populations, but did permit the identification of most of the prokaryotic components of the symbiotic system. Finally, fungi and yeasts were identified by both methods. Owing to the lack of sequenced genes in flagellates, targeting tubulins for LC-MS/MS could allow fingerprints of flagellate populations to be established. Experimental and technical improvements might increase the efficiency of identification of prokaryotic populations in the near future, based on metaproteomic development. [less ▲]

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See detailVolatile compounds of the traditional sorghum beers “ikigage” brewed with Vernonia amygdalina “umubirizi”
Lyumugabe, F.; Bajyana Songa, E.; Wathelet, Jean-Paul ULg et al

in Cerevisia : Belgian Journal of Brewing and Biotechnology (2013), 37

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See detailEffect of humic substances on in vitro rooting of alder (Alnus glutinosa (L.) Gaertn) and birch (Betula pendula Roth)
Tahiri, Abdelghani ULg; Destain, Jacqueline ULg; Druart, Philippe et al

Poster (2013)

Humic substances (HS) are organic compounds resulting from the physical, chemical and microbiological transformations of organic residues. Present everywhere in the nature; they are taking part in basic ... [more ▼]

Humic substances (HS) are organic compounds resulting from the physical, chemical and microbiological transformations of organic residues. Present everywhere in the nature; they are taking part in basic functionalities in any ecosystems involving soils, sediments, water and landfills. They are heterogeneous and complex carbon macromolecules. Our study aims to compare the effect of HS from différent sources on in vitro rooting of woody plants. [less ▲]

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See detailPhysicochemical characterization of dextrins prepared with amylases from sorghum malt
Ba, Khady; Blecker, Christophe ULg; Danthine, Sabine ULg et al

in Stärke = Starch (2013)

Dextrins are partially hydrolyzed starch products which are used in a wide range of applications. Hydrolysis can be performed by acid or enzymatic reaction. However the properties of dextrins are ... [more ▼]

Dextrins are partially hydrolyzed starch products which are used in a wide range of applications. Hydrolysis can be performed by acid or enzymatic reaction. However the properties of dextrins are influenced by the type of reaction and the source of starch. The aim of the present study was to determine physicochemical properties of dextrins obtained from hydrolysis of starches (corn and wheat) and flours (wheat and cassava) by using sorghum malt amylases. Hydrolysis of starches and flours was performed at 65°C, the hydrolysates were centrifuged and spray-dried for analysis. Physicochemical and structure of the powders were investigated. Results showed that the proportion of DP2 (maltose and isomaltose) was higher in dextrins and more in dextrins from wheat flour. Low water activity and low dissolution time were found in all dextrins. XRD patterns revealed that the crystalline structure partially disappeared in some of them. At 30% concentration, we noted a Newtonian behavior for the dextrins. The freeze-concentrated T0 g and the peak melting temperature (T0 m) determined by DSC (DSC) showed lowest values for dextrins from wheat. Thermogravimetric analysis (TGA) revealed that the dextrins were quite stable until 200°C. [less ▲]

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See detailDegradation of p-nitrophenol and bacteria with TiO2 xerogels sensitized in situ with tetra(4-carboxyphenyl)porphyrins
Tasseroul, Ludivine ULg; Lambert, Stéphanie ULg; Eskenazi, David ULg et al

in Journal of Photochemistry and Photobiology A : Chemistry (2013), 272

TiO2-based materials doped with porphyrins have been prepared using a sol-gel process. To stabilize the TiO2-dye interactions, free metal tetra(4-carboxyphenyl)porphyrin and nickel tetra(4-carboxyphenyl ... [more ▼]

TiO2-based materials doped with porphyrins have been prepared using a sol-gel process. To stabilize the TiO2-dye interactions, free metal tetra(4-carboxyphenyl)porphyrin and nickel tetra(4-carboxyphenyl) porphyrin were introduced in situ into the TiO2 matrix during the sol-gel process rather than by grafting. Samples were thoroughly characterized by TEM, X-ray diffraction, FT-IR, DR-UV/vis and their texture has been examined by nitrogen adsorption-desorption at 77 K. The low thermal treatment allows obtaining anatase-TiO2 particles with high specific surface area without porphyrin degradation. A significant improvement of the photoactivity under visible light of porphyrin doped-TiO2 xerogels was observed for p-nitrophenol and for bacterial degradation. © 2013 Elsevier B.V. All rights reserved. [less ▲]

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See detailA low-cost, multiplexable, automated flow cytometry procedure for the characterization of microbial stress dynamics in bioreactors
Brognaux, Alison ULg; Han, Shanshan; Sorensen, Soren J et al

in Microbial Cell Factories (2013), 12(100), 1-14

Background Microbial cell population heterogeneity is now recognized as a major source of issues in the development and optimization of bioprocesses. Even if single cell technologies are available for the ... [more ▼]

Background Microbial cell population heterogeneity is now recognized as a major source of issues in the development and optimization of bioprocesses. Even if single cell technologies are available for the study of microbial population heterogeneity, only a few of these methods are available in order to study the dynamics of segregation directly in bioreactors. In this context, specific interfaces have been developed in order to connect a flow cytometer directly to a bioreactor for automated analyses. In this work, we propose a simplified version of such an interface and demonstrate its usefulness for multiplexed experiments.Results A low-cost automated flow cytometer has been used in order to monitor the synthesis of a destabilized Green Fluorescent Protein (GFP) under the regulation of the fis promoter and propidium iodide (PI) uptake. The results obtained showed that the dynamics of GFP synthesis are complex and can be attributed to a complex set of biological parameters, i.e. on the one hand the release of protein into the extracellular medium and its uptake modifying the activity of the fis promoter, and on the other hand the stability of the GFP molecule itself, which can be attributed to the protease content and energy status of the cells. In this respect, multiplexed experiments have shown a correlation between heat shock and ATP content and the stability of the reporter molecule. Conclusion This work demonstrates that a simplified version of on-line FC can be used at the process level or in a multiplexed version to investigate the dynamics of complex physiological mechanisms. In this respect, the determination of new on-line parameters derived from automated FC is of primary importance in order to fully integrate the power of FC in dedicated feedback control loops. [less ▲]

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See detailMunkoyo : des racines comme sources potentielles en enzymes amylolytiques et une boisson fermentée traditionnelle (synthèse bibliographique)
Foma, Roland Kibwega; Destain, Jacqueline ULg; Kayisu, Kalenga et al

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (2013), 17(2), 352-363

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See detailComparison of plant defense elicitor properties of amphiphilic compounds from plant-associated bacteria
Mariutto, M.; Fernandez, O.; Thonart, Philippe ULg et al

in Bulletin OILB/SROP = IOBC/WPRS Bulletin (2013), 89

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See detailBiocontrol and Plant Growth Promotion Characterization of Bacillus Species Isolated from Calendula officinalis Rhizosphere
Ait Kaki, A.; Kacem Chaouche, N.; Dehimat, L. et al

in Indian Journal of Microbiology (2013)

The phenotypic and genotypic diversity of the plant growth promoting Bacillus genus have been widely investigated in the rhizosphere of various agricultural crops. However, to our knowledge this is the ... [more ▼]

The phenotypic and genotypic diversity of the plant growth promoting Bacillus genus have been widely investigated in the rhizosphere of various agricultural crops. However, to our knowledge this is the first report on the Bacillus species isolated from the rhizosphere of Calendula officinalis. 15 % of the isolated bacteria were screened for their important antifungal activity against Fusarium oxysporum, Botrytis cinerea, Aspergillus niger, Cladosporium cucumerinium and Alternaria alternata. The bacteria identification based on 16S r-RNA and gyrase-A genes analysis, revealed strains closely related to Bacillus amyloliquefaciens, B. velezensis, B. subtilis sub sp spizezenii and Paenibacillus polymyxa species. The electro-spray mass spectrometry coupled to liquid chromatography (ESI-LC MS) analysis showed that most of the Bacillus isolates produced the three lipopeptides families. However, the P. polymyxa (18SRTS) didn't produce any type of lipopeptides. All the tested Bacillus isolates produced cellulase but the protease activity was observed only in the B. amyloliquefaciens species (9SRTS). The Salkowsky colorimetric test showed that the screened bacteria synthesized 6-52 μg/ml of indole 3 acetic acid. These bacteria produced siderophores with more than 10 mm wide orange zones on chromazurol S. The greenhouse experiment using a naturally infested soil with Sclerotonia sclerotiorum showed that the B. amyloliquefaciens (9SRTS) had no significant (P > 0.05) effect on the pre-germination of the chickpea seeds. However, it increased the size of the chickpea plants and reduced the stem rot disease (P < 0.05).These results suggested that the Bacillus strains isolated in this work may be further used as bioinoculants to improve the production of C. officinalis and other crop systems. © 2013 Association of Microbiologists of India. [less ▲]

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See detailInfluence of lignin in Reticulitermes santonensis: symbiotic interations investigated through proteomics
Bauwens, Julien ULg; Tarayre, Cédric ULg; Brasseur, Catherine ULg et al

in Symbiosis (2013)

The gut of lower termites is populated by numerous microbial species belonging to prokaryotes, fungi, yeasts and protists. These micro-organisms are organized in a complex symbiotic system, interacting ... [more ▼]

The gut of lower termites is populated by numerous microbial species belonging to prokaryotes, fungi, yeasts and protists. These micro-organisms are organized in a complex symbiotic system, interacting together and with the insect host. Their likely ability to degrade ligno-cellulosic compounds could lead to improvements in second generation biofuels production. Lignin elimination represents a critical point as this polymer significantly interferes with industrial process of cellulose. Although host produces its own lignin-degrading enzymes, some symbionts may participate in digestion of lignin and its degradation products in termite gut. Here, we compared gut proteomes from R. santonensis after rearing on artificial diets composed of cellulose with and without lignin. The effect of lignin in artificial diets on different parts of the digestive tract was compared through liquid chromatography associated with tandem mass spectrometry (LC-MS/MS) experiments. Enzymatic assays were performed to characterize activities present in R. santonensis digestive tract after feeding on artificial diets. Microscopic observations of microbial communities provided some information on population balances after feeding experiment. [less ▲]

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See detailReal-time monitoring of cell viability and cell density on the basis of a three dimensional optical reflectance method (3D-ORM): investigation of the effect of sub-lethal and lethal injuries
Brognaux, Alison ULg; Bugge, Jörg; Schwartz, Friedel et al

in Journal of Industrial Microbiology & Biotechnology (2013)

Cell density and cell viability have been followed on-line by using a three-dimensional optical reflectance method (3D-ORM) probe. This method has allowed to highlight the differences between a well-mixed ... [more ▼]

Cell density and cell viability have been followed on-line by using a three-dimensional optical reflectance method (3D-ORM) probe. This method has allowed to highlight the differences between a well-mixed and a scale-down bioreactor configured in order to reproduce mixing deficiencies during a fed-batch culture of E. coli. These differences have been observed both for the obscuration factor (OBF) and the coincidence probability (COP) delivered by the probe. These parameters are correlated to flow cytometry measurement based on the PI-uptake test and cell density based on optical density measurement. This first set of results has pointed out the fact that the 3D-ORM probe is sensitive to sub-lethal injuries encountered by microbial cells in process-related conditions. The effect of lethal injuries has been further investigated on the basis of additional experiments involving heat stress and a sharp increase of the OBF has been observed indicating that cells are effectively injured by the increase of temperature. However, further improvement of the probe are needed in order to give access to single-cell measurements. [less ▲]

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See detailDirect and indirect use of GFP whole cell biosensors for the assessment of bioprocess performances: design of milliliter scale-down bioreactors
Brognaux, Alison ULg; Neubauer, Peter; Twizere, Jean-Claude ULg et al

in Biotechnology Progress (2013), 29(1), 48-59

Substrate limitation responsive biosensors have been used for the development of a mini-bioreactor platform that can be used as a scale-down tool. Three green fluorescent protein (GFP) transcriptional ... [more ▼]

Substrate limitation responsive biosensors have been used for the development of a mini-bioreactor platform that can be used as a scale-down tool. Three green fluorescent protein (GFP) transcriptional reporters have been chosen in Escherichia coli, i.e., uspA::gfp, csiE::gfp and yciG::gfp. Our previous studies have shown that these kinds of promoters are induced in response to substrate limitation and are significantly repressed when cultures are carried out in heterogeneous bioreactors. This sensitivity to substrate limitation has been confirmed in the case of the csiE and yciG biosensors. A mini-scale-down platform is proposed as a high throughput tool to rapidly investigate the usefulness of a given microbial biosensor. This platform is composed of shake flasks able to operate in fed-batch mode either using the slow release or the intermittent feeding principle. Local heterogeneities were reproduced at the level of these mini-bioreactors (operating under the intermittent feeding principle) and caused a decrease in GFP expression as in conventional scale-down reactors. The presence of GFP in supernatants was also noted and seems to be correlated with the substrate limitation signal for the three cultivation systems considered in this work (i.e., chemostat, conventional and mini-bioreactors) and with membrane permeability. [less ▲]

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See detailHydrolysis of starches and flours by sorghum malt amylases for dextrins production
Ba, Khady ULg; Aguedo, Mario ULg; Tine, Emmanuel et al

in European Food Research & Technology (2013)

Corn and wheat starches as well as wheat and cassava flours were hydrolyzed using sorghum malt at 65 °C for 6 hours. During these reactions, dextrose equivalent (DE) values were followed under 3 ... [more ▼]

Corn and wheat starches as well as wheat and cassava flours were hydrolyzed using sorghum malt at 65 °C for 6 hours. During these reactions, dextrose equivalent (DE) values were followed under 3 concentrations of sorghum malt and of calcium chloride. Wheat flour presented the highest DE values and cassava flour had the highest hydrolysis yield. Thus, different dextrins were produced in a pilot plant and were analyzed by HPSEC and HPAEC-PAD for their molecular weight distribution and oligosaccharides composition respectively. The results indicated that oligosaccharides with broad molecular weight distributions were present in the dextrins produced and that the proportion of maltose was very high. [less ▲]

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See detailFermentation of Kivuguto, a Rwandese traditional milk : selection of microbes for a starter culture
Karenzi, E.; Dubois-Dauphin, R.; Mashaku, A. et al

in Sciences & technologie C (2012), 36

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See detailUtilisation des termites comme source de microorganismes dans la filière de production du bioéthanol de seconde génération
Tarayre, Cédric ULg; Bauwens, Julien ULg; Brasseur, Catherine ULg et al

Poster (2012, November 14)

Les termites abritent une microflore symbiotique qui intervient dans la dégradation des fibres constitutives du bois, synthétisant des enzymes capables d’hydrolyser ses composants. Les sucres ... [more ▼]

Les termites abritent une microflore symbiotique qui intervient dans la dégradation des fibres constitutives du bois, synthétisant des enzymes capables d’hydrolyser ses composants. Les sucres fermentescibles libérés suite à cette hydrolyse sont utilisables dans le cadre de la production du bioéthanol de seconde génération. [less ▲]

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See detailImplementation of structured metal packing for the design of biofilm reactor : analysis by high energy X-ray tomography and application to the production of lipopeptides by Bacillus subtilis
Zune, Quentin ULg; Soyeurt, Delphine; Ongena, Marc ULg et al

Poster (2012, October 08)

1. Whereas multi-species biofilm reactors are commonly used for treatments of water and gas effluents, new strategies are arising for the development of mono-species biofilm reactors in order to produce ... [more ▼]

1. Whereas multi-species biofilm reactors are commonly used for treatments of water and gas effluents, new strategies are arising for the development of mono-species biofilm reactors in order to produce high added value molecules. Thus, it is required to design new bioreactors able to promote the growth of the biomass on the form of a biofilm and to identify the key physico-chemical parameters involved in order to optimize the bioprocess. 2. Aim of this study was to investigate a pilot-scale biofilm reactor comprising a metal structured packing promoting growth of Bacillus subtilis as a biofilm for the production of lipopeptides, high added value compounds with high surface active properties. 3. In this work, the mechanical stirring system of a 20L stirred tank bioreactor has been removed and replaced by a metal structured packing positioned in the headspace of the vessel above a liquid phase. The culture medium is continuously recirculated on the packing thanks to a peristaltic pump and air supply is performed just above the liquid phase under the packing. High energy X-ray tomography was used to estimate non-invasively the biofilm distribution inside the packing and permitted to define parameters that affect scale-up. Performances of the biofilm reactor were compared with a submerged culture in a stirred tank reactor in terms of lipopeptides production. 4. After 72 hours of fermentation, 94 % of the total biomass adheres onto the metal packing on the form of a biofilm. The colonization of this latter has been visualized non-invasively by X-ray tomography directly inside the packing and shows a conical repartition of the biofilm mass (about 25% of the total volume of the packing) as well as the presence of clogging. However, unlike the submerged culture, no foam formation appeared during fermentation and surfactin yield reaches 345,4 ± 32,8 mg / L for the biofilm reactor against 277,3 ± 34,4 mg / L in the stirred tank reactor. 5. In conclusion, this experimental setting leads to a major technological progress avoiding foam formation and increasing surfactin production. Nevertheless, significant improvements are required at the level of the biofilm distribution in thin layers inside the packing in order to increase mass transfer and lipopeptides recoveries. Further investigations will be devoted to the optimization of the physico-chemical parameters involved in biofilm distribution. [less ▲]

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See detailImplementation of structured metal packing for the design of biofilm reactor : analysis by high energy X-ray tomography and application to the production of lipopeptides by Bacillus subtilis
Zune, Quentin; Soyeurt, Delphine; Ongena, Marc ULg et al

Poster (2012, October 08)

1. Whereas multi-species biofilm reactors are commonly used for treatments of water and gas effluents, new strategies are arising for the development of mono-species biofilm reactors in order to produce ... [more ▼]

1. Whereas multi-species biofilm reactors are commonly used for treatments of water and gas effluents, new strategies are arising for the development of mono-species biofilm reactors in order to produce high added value molecules. Thus, it is required to design new bioreactors able to promote the growth of the biomass on the form of a biofilm and to identify the key physico-chemical parameters involved in order to optimize the bioprocess. 2. Aim of this study was to investigate a pilot-scale biofilm reactor comprising a metal structured packing promoting growth of Bacillus subtilis as a biofilm for the production of lipopeptides, high added value compounds with high surface active properties. 3. In this work, the mechanical stirring system of a 20L stirred tank bioreactor has been removed and replaced by a metal structured packing positioned in the headspace of the vessel above a liquid phase. The culture medium is continuously recirculated on the packing thanks to a peristaltic pump and air supply is performed just above the liquid phase under the packing. High energy X-ray tomography was used to estimate non-invasively the biofilm distribution inside the packing and permitted to define parameters that affect scale-up. Performances of the biofilm reactor were compared with a submerged culture in a stirred tank reactor in terms of lipopeptides production. 4. After 72 hours of fermentation, 94 % of the total biomass adheres onto the metal packing on the form of a biofilm. The colonization of this latter has been visualized non-invasively by X-ray tomography directly inside the packing and shows a conical repartition of the biofilm mass (about 25% of the total volume of the packing) as well as the presence of clogging. However, unlike the submerged culture, no foam formation appeared during fermentation and surfactin yield reaches 345,4 ± 32,8 mg / L for the biofilm reactor against 277,3 ± 34,4 mg / L in the stirred tank reactor. 5. In conclusion, this experimental setting leads to a major technological progress avoiding foam formation and increasing surfactin production. Nevertheless, significant improvements are required at the level of the biofilm distribution in thin layers inside the packing in order to increase mass transfer and lipopeptides recoveries. Further investigations will be devoted to the optimization of the physico-chemical parameters involved in biofilm distribution. [less ▲]

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