References of "Lejeune, Annick"
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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 detailModélisation et étude expérimentale du comportement de Saccharomyces cerevisiae (boulardii) lors de l'extrapolation du procédé à des bioréacteurs de volume industriel
Lejeune, Annick ULg

Doctoral thesis (2014)

L’extrapolation de la production d’un microorganisme est un problème complexe qui prend en compte de nombreux paramètres. En effet, les paramètres opératoires induisent des conditions environnementales ... [more ▼]

L’extrapolation de la production d’un microorganisme est un problème complexe qui prend en compte de nombreux paramètres. En effet, les paramètres opératoires induisent des conditions environnementales qui peuvent être dommageables pour les cellules se développant dans le réacteur. Il est donc important de prendre en compte le côté génie chimique pour comprendre quelles sont les conditions hydrodynamiques présentes dans le réacteur et ensuite s’attacher à étudier la réponse des cellules vis-à-vis de ces conditions. Ce travail se place dans le cas particulier de la production de biomasse de Saccharomyces cerevisiae (boulardii). Dans ce cas, le point critique est l’ajout de la solution concentrée de substrat dans le réacteur. L’utilisation de réacteurs scale-down a permis d’étudier différents cas de conditions hydrodynamiques défavorables, ainsi que leurs impacts sur différents paramètres. Dans un premier temps, l’étude des paramètres ségrégés (concentration en biomasse, en co-produits…) a montré, notamment, une diminution du rendement et une augmentation de la concentration en éthanol. Ensuite, l’étude s’est placée au niveau cellulaire avec l’utilisation de souches exprimant une protéine recombinante fluorescente et de marqueurs cellulaires fluorescents, couplée à la cytométrie en flux. Ces techniques ont permis de mettre en évidence la présence de différentes sous-populations cellulaires, ainsi que l’apparition du phénomène de résistance cellulaire aux stress lors de la production en réacteurs scale-down. L’importance du choix de la méthode de traitement des données cytométriques a également été débattue. Ce travail a abordé différents domaines pour tenter de mieux comprendre la réponse cellulaire aux fluctuations des conditions environnementales ressenties par Saccharomyces cerevisiae (boulardii) lors de productions en réacteurs mimant les conditions hydrodynamiques des réacteurs de grands volumes. [less ▲]

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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 detailStress microbien lors du processus d’extrapolation: Approche physique et biologique
Lejeune, Annick ULg; Thonart, Philippe ULg; Delvigne, Frank ULg

in Cerevisia : Belgian Journal of Brewing and Biotechnology (2013), 38(3), 89-101

Les micro-organismes sont utilisés dans de nombreux domaines (agro-alimentaire, pharmaceutique, environnemental, énergétique,…) que ce soit pour la production de biomasse ou de métabolites particuliers ... [more ▼]

Les micro-organismes sont utilisés dans de nombreux domaines (agro-alimentaire, pharmaceutique, environnemental, énergétique,…) que ce soit pour la production de biomasse ou de métabolites particuliers. Le passage de l’échelle du laboratoire à l’échelle industrielle est souvent problématique car les micro-organismes sont sensibles aux conditions environnementales développées au sein du volume réactionnel. De plus, la perte de l’efficacité d’homogénéisation des bioréacteurs industriels entraîne des perturbations au niveau du métabolisme des cellules. C’est pourquoi il est important d’étudier les conditions hydrodynamiques développées au sein du réacteur. Cet article présente des méthodes de calcul et de modélisation de ces conditions. L’impact de l’hydrodynamique sur le métabolisme microbien peut être étudié par l’utilisation de réacteurs scale-down. Les techniques nouvelles permettant de réaliser un scale-up prennent en compte des caractères physiologiques des cellules, qui estiment l’impact des conditions environnementales sur le métabolisme microbien et donc la bonne réussite du procédé. Enfin, l’étude de Saccharomyces cerevisiae, micro-organisme de grand intérêt biotechnologique, est réalisée. [less ▲]

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See detailPhysiological response of yeast to process perturbations: A mini-bioreactor approach
Lejeune, Annick ULg; Delvigne, Frank ULg; Thonart, Philippe ULg

in Cerevisia : Belgian Journal of Brewing and Biotechnology (2013), 38(1), 15-19

Large-scale production of yeast (Saccharomyces cerevisiae) is difficult to control, considering the drop of mixing and mass transfer efficiency during scale-up. The drop of hydrodynamic efficiency in ... [more ▼]

Large-scale production of yeast (Saccharomyces cerevisiae) is difficult to control, considering the drop of mixing and mass transfer efficiency during scale-up. The drop of hydrodynamic efficiency in large-scale bioreactors induces the formation of heterogeneities, i.e. mainly substrate and dissolved oxygen in process conditions. These extracellular fluctuations have several impacts at the level of the physiology of microorganisms, from metabolic shift to specific gene expression (stress response). Microbial cell responses to extracellular fluctuations are actually not fully understood. In this work, we propose to reproduce the main extracellular fluctuations at the level of a mini bioreactor platform. These mini-reactors are shake flasks equipped with dissolved oxygen probes. The cultures are realized with different fed-batch control strategies. A scale-down approach has been developed by considering slow release techniques and intermittent feeding, in order to reproduce the glucose and dissolved oxygen fluctuations experienced in large-scale reactors. The mini-reactor has been used to screen the response of several green fluorescent protein (GFP) reporter strains (adh2, tps2, pdc6 and hxt2). The GFP content of cells has been determined by flow cytometry in order to take into account population heterogeneity. In front of these results, the methodology presented in this work can be proposed as a scale-down tool. [less ▲]

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See detailL'extrapolation des bioréacteurs : un problème de génie des procédés ou de physiologie microbienne ?
Delvigne, Frank ULg; Destain, Jacqueline ULg; Brognaux, Alison ULg et al

in Chimie Nouvelle (2010), 104

Bioreactor scale-up often pose a serious issue during the industrial development of a bioprocess considering the numerous physical and biological phenomena occurring in the reacting volume. The basic ... [more ▼]

Bioreactor scale-up often pose a serious issue during the industrial development of a bioprocess considering the numerous physical and biological phenomena occurring in the reacting volume. The basic principles of scale-up coming from the traditional chemical and process engineering approaches will be first reviewed and will be then compared to a new one involving recent development at the level of microbial strain manipulation. This "physiological" approach of scale-up involves directly a biological component of the system (by comparison with the traditional approach for scaling-up involving physical parameters indirectly linked to the physiological phenomena occurring in the bioreactor), i.e. the synthesis of a reporter fluorescent protein when microbial cells are exposed to stress. It will be shown how this principle can be used for a better understanding of the relationship between bioreactor hydrodynamics and microbial stress. [less ▲]

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See detailInfluence of bioreactor hydraulic characteristics on a Saccharomyces cerevisiae fed-batch culture: hydrodynamic modelling and scale-down investigations.
Lejeune, Annick ULg; Delvigne, Frank ULg; Thonart, Philippe ULg

in Journal of Industrial Microbiology & Biotechnology (2010), 37

Yeast is a widely used microorganism at the industrial level because of its biomass and metabolite production capabilities. However, due to its sensitivity to the glucose effect, problems occur during ... [more ▼]

Yeast is a widely used microorganism at the industrial level because of its biomass and metabolite production capabilities. However, due to its sensitivity to the glucose effect, problems occur during scale-up to the industrial scale. Hydrodynamic conditions are not ideal in large-scale bioreactors, and glucose concentration gradients can arise when these bioreactors are operating in fed-batch mode. We have studied the effects of such gradients in a scale-down reactor, which consists of a mixed part linked to a non-mixed part by a recirculation pump, in order to mimic the hydrodynamic conditions encountered at the large scale. During the fermentation tests in the scale-down reactor, there was a drop in both biomass yield (ratio between the biomass produced and the glucose added) and trehalose production and an increase in both fermentation time (time between inoculation and beginning of stationary phase) and ethanol production. We have developed a stochastic model which explains these effects as the result of an induction process determined mainly by the hydrodynamic conditions. The concentration profiles experienced by the microorganisms during the scale-down tests were expressed and linked to the biomass yields of the scale-down tests. [less ▲]

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See detailTrehalose as a stress marker of the physiological impact of mixing on yeast production: scale-down reactors and mini-bioreactors investigations
Lejeune, Annick ULg; Delvigne, Frank ULg; Thonart, Philippe ULg

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

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See detailStochastic models to study the impact of mixing on a fed-batch culture of Saccharomyces cerevisiae.
Delvigne, Frank ULg; Lejeune, Annick ULg; Destain, Jacqueline ULg et al

in Biotechnology Progress (2006), 22(1), 259-69

The mechanisms of interaction between microorganisms and their environment in a stirred bioreactor can be modeled by a stochastic approach. The procedure comprises two submodels: a classical stochastic ... [more ▼]

The mechanisms of interaction between microorganisms and their environment in a stirred bioreactor can be modeled by a stochastic approach. The procedure comprises two submodels: a classical stochastic model for the microbial cell circulation and a Markov chain model for the concentration gradient calculus. The advantage lies in the fact that the core of each submodel, i.e., the transition matrix (which contains the probabilities to shift from a perfectly mixed compartment to another in the bioreactor representation), is identical for the two cases. That means that both the particle circulation and fluid mixing process can be analyzed by use of the same modeling basis. This assumption has been validated by performing inert tracer (NaCl) and stained yeast cells dispersion experiments that have shown good agreement with simulation results. The stochastic model has been used to define a characteristic concentration profile experienced by the microorganisms during a fermentation test performed in a scale-down reactor. The concentration profiles obtained in this way can explain the scale-down effect in the case of a Saccharomyces cerevisiae fed-batch process. The simulation results are analyzed in order to give some explanations about the effect of the substrate fluctuation dynamics on S. cerevisiae. [less ▲]

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