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See detailStochastic exposure to sub-lethal high temperature enhances exopolysaccharides (EPS) excretion and improves Bifidobacterium bifidum cell survival to freeze-drying
Nguyen, Huu Thanh ULg; Razafindralambo, Hary; Blecker, Christophe ULg et al

in Biochemical Engineering Journal (2014), 88

Exposure of microbial cells to sub-lethal stresses is known to increase cell robustness. In this work, a two-compartment bioreactor in which microbial cells are stochastically exposed to sub-lethal ... [more ▼]

Exposure of microbial cells to sub-lethal stresses is known to increase cell robustness. In this work, a two-compartment bioreactor in which microbial cells are stochastically exposed to sub-lethal temperature stresses has been used in order to investigate the response of the stress sensitive Bifidobacterium bifidum THT 0101 to downstream processing operations. A stochastic model validated by residence time distribution experiments has shown that in the heat-shock configuration, a two-compartment bioreactor (TCB) allows the exposure of microbial cells to sub-lethal temperature of 42°C for a duration comprised between 100 and 300 seconds. This exposure resulted in a significant increase of cell resistance to freeze-drying by comparison with cells cultivated in conventional bioreactors or in the TCB in the cold shock mode (CS-TCB). The mechanism behind this robustness seems to be related with the coating of microbial cells with exopolysaccharide (EPS), as assessed by the change of the zeta potential and the presence of higher EPS concentration after heat shock. Conditioning of Bifidobacteria on the basis of the heat shock technique is interesting from the practical and economical point of view since this strategy can be directly implemented in the bioreactor during stationary phase preceding cell recovery and freeze-drying [less ▲]

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See detailEnzymatic process for the fractionation of baker's yeast cell wall (saccharomyces cerevisiae)
Borchani, Chema; Fonteyn, Fabienne; Jamin, Guilhem et al

Conference (2014, April 07)

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See detailImaging MS: strategies for the identification of analytes
Debois, Delphine ULg; Smargiasso, Nicolas ULg; Jourdan, Emmanuel et al

Scientific conference (2014, April 04)

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See detailUse of on-line flow cytometry for the characterization of microbial stress dynamics during the bioprocess
Brognaux, Alison ULg; Han, Shanshan; Sorensen, Soren et al

Conference (2014, April 03)

Microbial cell population heterogeneity is now recognized as a major source of issues for the development and optimization of bioprocesses. Flow cytometry is a very powerful tool for the follow up of ... [more ▼]

Microbial cell population heterogeneity is now recognized as a major source of issues for the development and optimization of bioprocesses. Flow cytometry is a very powerful tool for the follow up of physiological properties of microbial cells in process-related conditions at the single cell level, and can be used to study the dynamics of segregation directly in bioreactors. In this context, specific interfaces have been developed in order to connect flow cytometer (FC) directly on bioreactor for automated analyses. In this work, we propose a simplified version of such interface and demonstrated its usefulness for multiplexed experiments. This automated FC system has been tested for the follow up of the dynamics of an E. coli pfis::gfpAAV fluorescent bio-reporter and its PI uptake, correlated with membrane permeability. This bioreporter is composed of a fis promoter, a growth dependent promoter-indicator of the nutrient status of cells, fused to a gene expressing an unstable variant of GFP. The results obtained showed that the dynamics of the GFP synthesis is complex and can be attributed to a complex set of biological parameters. Segregation in the membrane permeability has been noticed. This work demonstrates that a simplified version of on-line FC can be used at the process level for the investigation of the dynamics of complex physiological mechanisms. [less ▲]

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See detailCFD-based Compartment model for description of mixing in bioreactors
Delafosse, Angélique ULg; Calvo, Sébastien ULg; Collignon, Marie-Laure ULg et al

in Chemical Engineering Science (2014), 106

In most bioprocesses, it is fundamental to accurately predict the hydrodynamics behavior of bioreactors of different size and its interaction with the biological reaction. Computational Fluid Dynamics can ... [more ▼]

In most bioprocesses, it is fundamental to accurately predict the hydrodynamics behavior of bioreactors of different size and its interaction with the biological reaction. Computational Fluid Dynamics can provide detailed modeling about hydrodynamics and mixing. However, it is computationally intensive, especially when reactions are taken into account. Another way to predict hydrodynamics is the use of “Compartment” or “Network-of-zones” model which are much less demanding in computation time than CFD. However, compartments and fluxes between them are often defined by considering global quantities not representative of the flow complexity. To overcome the limitations of these two methods, a solution is to combine compartment modeling and CFD simulations. The aim of this study is to propose a compartment model where the flow rates between two adjacent compartments are easily computed from the velocity fields obtained by CFD. The mixing evolution predicted by the CFD-based compartment model have been then compared with mixing experiment results. Unlike a CFD mixing simulation and a classical compartment model, the CFD-based compartment model proposed in this work reproduces with a good accuracy the spatial distribution of concentrations during the mixing process and this, without any adjustable parameters. [less ▲]

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See detailScale-down effect on the extracellular proteome of Escherichia coli: correlation with membrane permeability and modulation according substrate heterogeneities
Brognaux, Alison ULg; Francis, Frédéric ULg; Twizere, Jean-Claude ULg et al

in Bioprocess and Biosystems Engineering (2014)

Protein leakage is induced in well-mixed fed-batch bioreactor by comparison with cultures carried out in scale-down conditions. This effect is attributed to a progressive increase of cell membrane ... [more ▼]

Protein leakage is induced in well-mixed fed-batch bioreactor by comparison with cultures carried out in scale-down conditions. This effect is attributed to a progressive increase of cell membrane permeability and the synthesis of several outer-membrane components allowing to cope with substrate limitation commonly found in high-cell density culture. A comparative analysis of protein leakage has thus been performed in well-mixed bioreactors and in scale-down devices. The extracellular proteome of E.coli has been investigated by 2D-gel electrophoresis and identified by subsequent MALDI-TOF analysis. On 110 picked spots, 67 proteins have been identified and the sub-localisation and the molecular function of these proteins have been determined. A majority of the extracellular proteome was composed of outer-membrane and periplasmic proteins (64%) confirming the fact that leakage is involved in high-cell density cultures. About 50% of this extracellular proteome was composed of transport and binding proteins. Furthermore, the more abundant spots on the gel corresponded to porin proteins and periplasmic transporters. In particular, the OmpC porin was found to be very abundant. Moreover, the scale-down effect on this extracellular proteome has been investigated by 2D-DIGE analysis (2-Dimensional Differential in-Gel Electrophoresis) and significant differences have been observed by comparison with culture carried out in well-mixed systems. Indeed, since substrate limitation signal is alleviated in this kind of apparatus, cell permeability was lowered as shown by flow cytometry. In scale-down conditions, protein leakage was thus less abundant. [less ▲]

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See detailScale-down effect on the extracellular proteome of Escherichia coli: correlation with membrane permeability and modulation according substrate heterogeneities
Brognaux, Alison ULg; Francis, Frédéric ULg; Twizere, Jean-Claude ULg et al

in Bioprocess and Biosystems Engineering (2014)

Protein leakage is induced in well-mixed fed-batch bioreactor by comparison with cultures carried out in scale-down conditions. This effect is attributed to a progressive increase of cell membrane ... [more ▼]

Protein leakage is induced in well-mixed fed-batch bioreactor by comparison with cultures carried out in scale-down conditions. This effect is attributed to a progressive increase of cell membrane permeability and the synthesis of several outer-membrane components allowing to cope with substrate limitation commonly found in high-cell density culture. A comparative analysis of protein leakage has thus been performed in well-mixed bioreactors and in scale-down devices. The extracellular proteome of E.coli has been investigated by 2D-gel electrophoresis and identified by subsequent MALDI-TOF analysis. On 110 picked spots, 67 proteins have been identified and the sub-localisation and the molecular function of these proteins have been determined. A majority of the extracellular proteome was composed of outer-membrane and periplasmic proteins (64%) confirming the fact that leakage is involved in high-cell density cultures. About 50% of this extracellular proteome was composed of transport and binding proteins. Furthermore, the more abundant spots on the gel corresponded to porin proteins and periplasmic transporters. In particular, the OmpC porin was found to be very abundant. Moreover, the scale-down effect on this extracellular proteome has been investigated by 2D-DIGE analysis (2-Dimensional Differential in-Gel Electrophoresis) and significant differences have been observed by comparison with culture carried out in well-mixed systems. Indeed, since substrate limitation signal is alleviated in this kind of apparatus, cell permeability was lowered as shown by flow cytometry. In scale-down conditions, protein leakage was thus less abundant. [less ▲]

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See detailEffect of encapsulated nanoparticles on thermophillic anaerobic digestion
Al-Ahmad, Alaa Eddin ULg; Hiligsmann, Serge ULg; Lambert, Stéphanie ULg et al

Poster (2014, February 07)

Recently, enormous interest has been focused on biological applications of metal nanoparticles NPs due to their small size, high specified surface and their great potential in application to many science ... [more ▼]

Recently, enormous interest has been focused on biological applications of metal nanoparticles NPs due to their small size, high specified surface and their great potential in application to many science fields. The most studied process concerns zero valent palladium and iron NPs improving anaerobic biodegradation of chlorinated hydrocarbons (Windt et al., 2005). Moreover, investigation carried out in our lab showed that iron NPs encapsulated in silicate matrix may enhance hydrogen production by Clostridium butyricum (Beckers et al., 2013). Nevertheless the influences of metal NPs on methane producing anaerobic digestion have seldom been investigated. The present work investigates the enhancement effect of seven different metal NPs on methane production during the thermophilic anaerobic digestion. NPs of Cu, Pd, Pt, Ni, Co, Ag and Fe encapsulated in porous silica (SiO2) to prevent their coagulation and agglomeration, were added at concentration of 10-5mol/L in batch test (125ml serum bottles containing 70mL culture medium with 5g/L acetate monohydrate as the sole carbon substrate). Nickel, cobalt and iron NPs improved methane production from acetate. To confirm the previous results, the NPs were tested at different concentrations (10-4, 10-5, and 10-6 mol/L) with starch and glucose substrates. The results show that the impact increases with the increase of NPs concentrations up to 10-4 mol/L. The modified Gompertz equation was applied to describe the effect of NPs on anaerobic digestion. According to this model, the kinetic of methane production was particularly affected by nanoparticles addition. The values of the maximum methane production rate MPR (ml/day) was significantly higher 72.5% with nickel NPs at a concentration of 10-4 mol/L than the control without NPs. [less ▲]

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See detailUse of on-line flow cytometry for the characterization of microbial stress dynamics during the bioprocess
Brognaux, Alison ULg; Han, Shanshan; Sorensen, Soren et al

Poster (2014, February 07)

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See detailNew alternatives to chemical pesticides: deciphering the action mechanisms of lipid based plant elicitors via complementary biophysical and biological approaches.
Nasir, Mehmet Nail ULg; Polo Lozano, Damien ULg; Luzuriaga Loaiza, Walter ULg et al

Poster (2014, February)

Nowadays, many health and environmental problems are caused by the use of chemical pesticides. In this context, an increasing demand for alternative products such as biopesticides has been observed. Among ... [more ▼]

Nowadays, many health and environmental problems are caused by the use of chemical pesticides. In this context, an increasing demand for alternative products such as biopesticides has been observed. Among biopesticides, elicitor molecules which are able to trigger immune defense responses in plants are one of the most promising options. Although numerous elicitors have been discovered, the mechanisms involved in the perception, by plants, of only a few molecules have been identified. These elicitors usually interact with proteic receptors but we have recently shown that they may also act on the lipid phase of the plasma membrane. This project first aims to improve our understanding of the molecular mechanisms involved in the recognition of specific lipid based elicitors (LBE). On that basis, the FIELD project will contribute to the design and the development of innovative compounds derived natural LBE. A multi-disciplinary approach, based on chemistry, bio-physics, bio-chemistry, and phytopathology will be followed by a consortium of different research groups from Gembloux Agro-Bio Tech in close collaboration with teams from foreign institutions. [less ▲]

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See detailThe role of protein modifications in senescence of freeze-dried Acetobacter senegalensis during storage
Shafiei, Rasoul ULg; Zarmehrkhorshid, Raziyeh ULg; Bentaib, Azeddine ULg et al

in Microbial Cell Factories (2014)

Background Loss of viability is one of the most important problems during starter culture production. Previous research has mostly focused on the production process of bacterial starters, but there are ... [more ▼]

Background Loss of viability is one of the most important problems during starter culture production. Previous research has mostly focused on the production process of bacterial starters, but there are few studies about cellular protein deterioration causing cell defectiveness during storage. In the present study, we investigated the influence of storage temperature (−21, 4, 35°C) on the cellular protein modifications which may contribute to the senescence of freeze-dried Acetobacter senegalensis. Results Heterogeneous populations composed of culturable cells, viable but non-culturable cells (VBNC) and dead cells were generated when freeze-dried cells were kept at −21 and 4°C for 12 months whereas higher storage temperature (35°C) mainly caused death of the cells. The analysis of stored cell proteome by 2D-DiGE demonstrated a modified pattern of protein profile for cell kept at 4 and 35°C due to the formation of protein spot trains and shift of Isoelectric point (pI). Quantification of carbonylated protein by ELISA showed that the cells stored at 4 and 35°C had higher carbonylated protein contents than fresh cells. 2D-DiGE followed by Western blotting also confirmed the carbonylation of cellular proteins involved in translation process and energy generation. The auto-fluorescent feature of cells kept at 35°C increased significantly which may be an indication of protein glycation during storage. In addition, the percentage of cellular unsaturated fatty acid and the solubility of cellular proteins decreased upon storage of cells at higher temperature suggesting that peroxidation of fatty acids and possibly protein lipidation and oxidation occurred. Conclusions High storage temperature induces some deteriorative reactions such as protein oxidation, lipidation and glycation which may cause further protein modifications like pI-shift, and protein insolubility. These modifications can partly account for the changes in cell viability. It can also be deduced that even moderate carbonylation of some critical cellular proteins (like ribosomal proteins) may lead to VBNC formation or death of freeze-dried bacteria. Moreover, it seems that other mechanisms of biomolecule deterioration preceding protein carbonylation lead to VBNC formation under very low storage temperature. [less ▲]

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See detailTechnologie de production de masse d’insectes - INSECTECH
Richard, Gaetan ULg; Hance, Thierry; Fauconnier, Marie-Laure ULg et al

Report (2014)

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See detailCharacterization of Lactobacillus strains isolated from Algerian children faeces for their probiotic properties
Bahri, F.; Lejeune, Annick ULg; Dubois-Dauphin, R. et al

in African Journal of Microbiology Research [=AJMR] (2014), 8(3), 297-303

Lactic acid bacteria termed probiotics have preventive as well as curative effects on several types of diarrhoea of different aetiologies. The main objective of this study was to screen lactobacilli ... [more ▼]

Lactic acid bacteria termed probiotics have preventive as well as curative effects on several types of diarrhoea of different aetiologies. The main objective of this study was to screen lactobacilli strains having probiotic traits, isolated from Algerian healthy children faeces on the purpose of using them further in children diarrheal illnesses. One hundred and twenty (120) lactic acid bacteria isolates were selected from faecal samples of healthy Algerian children aged between five and ten years. Gram positive rods and catalase negative bacteria (52 isolates) were screened, in vitro, for their probiotic potential properties including ability to survive in simulated gastro-intestinal conditions, adherence to Caco-2 cells and their antimicrobial activity. The results show that only five strains resisted in simulated gastric juice at pH 1.5 and pepsin. Four of them were resistant to simulated intestinal conditions at pH 8 and pancreatin and have a good adherence. In the end, three of them were retained as they display interesting probiotic profiles characterized by a strong antimicrobial effect against some intestinal pathogenic bacteria. They were identified by 16S rDNA sequencing as Lactobacillus plantarum F12, Lactobacillus brevis G6 and Lactobacillus paracasei B13. [less ▲]

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See detailIsolation of an amylolytic chrysophyte, Poterioochromonas sp. from the digestive tract of the termite R. santonensis
Tarayre, Cédric ULg; Bauwens, Julien ULg; Brasseur, Catherine ULg et al

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (2014), 18(1),

The aim of this work was the isolation and cultivation of amylolytic protists living in the digestive tract of the termite Reticulitermes santonensis (Feytaud). A chrysophyte identified as ... [more ▼]

The aim of this work was the isolation and cultivation of amylolytic protists living in the digestive tract of the termite Reticulitermes santonensis (Feytaud). A chrysophyte identified as Poterioochromonas sp. was isolated in a special medium containing rice grains as a source of carbon and nitrogen. Then, the protist was grown in a medium containing starch as a carbon source, tryptone, and a phosphate buffer at different pH values (5, 6 and 7). Yeast extract was added or not. Ciprofloxacin was used to avoid the bacterial development. Other antibiotics were also tested but showed an inhibitive effect on the growth of Poterioochromonas sp. Yeast extract allowed reaching 1.9 (pH 5), 2.3 (pH 6) and 2.2 (pH 7) times higher final cell concentrations, and 2.8 (pH 5), 2.8 (pH 6) and 2.2 (pH 7) times higher biomass yields. The starch concentration did not decrease in the medium until 3 and 4 days of culture, with and without yeast extract, respectively. Eight days of culture were necessary for hydrolyzing the starch completely, with and without yeast extract. Maltose and maltotriose were detected in the culture media and were hydrolyzed progressively. Maximal maltose concentrations were 0.68, 0.66 and 0.51 g.l-1 in the medium containing yeast extract. Maltotriose concentrations were only 0.17, 0.14 and 0.12 g.l-1. Other glucose oligomers were also detected but in lower quantities. It was determined that the protist developed a weak amylase activity, particularly at a weakly acidic pH (5-6). Such a pH also allowed a better growth of the protist. A maximal amylase activity of 112 nkat.l-1 was measured with yeast extract at pH 5. No other enzymatic activity (protease, cellulase or xylanase) was detected except amylase. The degradation products of starch which were obtained by enzymatic hydrolysis allow the identification of α-amylase, amyloglucosidase and possibly β-amylase activities. [less ▲]

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See detailLack of isocitrate lyase in Chlamydomonas leads to changes in carbon metabolism and in the response to oxidative stress under mixotrophic growth.
Plancke, Charlotte; Vigeolas, Hélène ULg; Hohner, Ricarda et al

in The Plant journal : for cell and molecular biology (2014), 77(3), 404-417

Isocitrate lyase is a key enzyme of the glyoxylate cycle. This cycle plays an essential role in cell growth on acetate, and is important for gluconeogenesis as it bypasses the two oxidative steps of the ... [more ▼]

Isocitrate lyase is a key enzyme of the glyoxylate cycle. This cycle plays an essential role in cell growth on acetate, and is important for gluconeogenesis as it bypasses the two oxidative steps of the tricarboxylic acid (TCA) cycle in which CO2 is evolved. In this paper, a null icl mutant of the green microalga Chlamydomonas reinhardtii is described. Our data show that isocitrate lyase is required for growth in darkness on acetate (heterotrophic conditions), as well as for efficient growth in the light when acetate is supplied (mixotrophic conditions). Under these latter conditions, reduced acetate assimilation and concomitant reduced respiration occur, and biomass composition analysis reveals an increase in total fatty acid content, including neutral lipids and free fatty acids. Quantitative proteomic analysis by 14 N/15 N labelling was performed, and more than 1600 proteins were identified. These analyses reveal a strong decrease in the amounts of enzymes of the glyoxylate cycle and gluconeogenesis in parallel with a shift of the TCA cycle towards amino acid synthesis, accompanied by an increase in free amino acids. The decrease of the glyoxylate cycle and gluconeogenesis, as well as the decrease in enzymes involved in beta-oxidation of fatty acids in the icl mutant are probably major factors that contribute to remodelling of lipids in the icl mutant. These modifications are probably responsible for the elevation of the response to oxidative stress, with significantly augmented levels and activities of superoxide dismutase and ascorbate peroxidase, and increased resistance to paraquat. [less ▲]

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See detailMicroorganism-associated semiochemicals reduce the size of aphid populations in potato fields
Alabi, Taofic ULg; Brostaux, Yves ULg; Grigorescu, Alina ULg et al

in Revue des Régions Arides (2014), 35

The chemical cues released by many insect species, including agricultural pests, are used by predators and parasitoids to locate their prey or host. For instance, aphids excrete honeydew, which contains ... [more ▼]

The chemical cues released by many insect species, including agricultural pests, are used by predators and parasitoids to locate their prey or host. For instance, aphids excrete honeydew, which contains bacteria that produce semiochemicals. Ladybeetles and hoverflies use these semiochemicals to locate the colonies of prey aphid species. One bacterium (Staphylococcus sciuri) has been identified in the honeydew of the pea aphid, Acyrthosiphon pisum. This bacterium is responsible for the production of kairomonal substances, which guide hoverflies to aphid colonies. In the present study, we cultivated S. sciuri, and used solid-phase microextraction (SPME) to confirm the ability of this bacterium to produce 3-methyl-2-butenal and 3-methyl-2-butenoic acid, which previous studies have demonstrated as being the two semiochemicals that exhibit kairomonal activity. We subsequently conducted field experiments to evaluate the efficiency of two solutions as biological products to control aphid populations inhabiting potato plants; the first solution contained a suspension of living S. sciuri, and the second solution contained a mixture of the two semiochemicals produced by this bacterium. While the semiochemical solution did not lead to a significant reduction in aphid number, potato plants treated with the S. sciuri solution were infested with 28% less aphids compared to untreated plants. This study demonstrates the potential of using naturally occurring bacteria as a form of biological control of aphid infestations in agricultural management. [less ▲]

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See detailWhole-genome sequence of Serratia symbiotica strain CWBI-2.3T, a free-living symbiont of the Black Bean Aphid Aphis fabae
Foray, Vincent; Grigorescu, Alina ULg; Sabri, Ahmed et al

in Genome Announcements (2014), 2(4),

The gammaproteobacterium Serratia symbiotica is one of the major secondary symbionts found in aphids. Here, we report the draft genome sequence of S. symbiotica strain CWBI-2.3T, previously isolated from ... [more ▼]

The gammaproteobacterium Serratia symbiotica is one of the major secondary symbionts found in aphids. Here, we report the draft genome sequence of S. symbiotica strain CWBI-2.3T, previously isolated from the black bean aphid Aphis fabae. The 3.58-Mb genome sequence might provide new insights to understand the evolution of insect-microbe symbiosis. [less ▲]

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