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| Reference : E.coli prpoS::gfp strain as biosensor of glucose heterogeneity inside industrial bioreac... |
| Scientific congresses and symposiums : Poster | |||
| Life sciences : Biotechnology | |||
| http://hdl.handle.net/2268/79346 | |||
| E.coli prpoS::gfp strain as biosensor of glucose heterogeneity inside industrial bioreactors | |
| English | |
Brognaux, Alison  [Université de Liège - ULg > > > Form. doct. sc. agro. & ingé. biol.] | |
| Delvigne, Frank [Université de Liège - ULg > Chimie et bio-industries > Bio-industries >] | |
| Thonart, Philippe [Université de Liège - ULg > Département des sciences de la vie > Biochimie et microbiologie industrielles - Bio-industries >] | |
| 8-Dec-2010 | |
| International | |
| Biowin Day 2010. "Science for business: open innovation, the new model to innovate in health" | |
| 8 décembre 2010 | |
| pôle de compétitivité Biowin | |
| Louvain-la-neuve | |
| Belgique | |
| [en] Flow cytometry ; scale-up ; stress gene | |
| [en] • OBJECTIVE: Escherichia coli is a microorganism widely used in the industry for the
production of recombinant proteins. The performances obtained at the laboratory level are not reproducible at a large scale. Actually, the mixing operation is not efficient enough: gradients of glucose and oxygen appear when operating in fed-batch mode (addition of glucose during the culture). These gradients cause adverse impacts on the production of biomass and recombinant protein. The aim of this work is to use the microbial population as biocaptor of the encoutered stress inside heterogeneous industrial bioreactors to better scale-up and regulate these reactors. • MATERIALS AND METHODS: A plasmid carrying a stress promoter followed by the coding sequence of the Green Fluorescent Protein (GFP) is introduced in the bacterial host (in our case, the strain E. coli K12 will be considered as a model organism). When the cell is submitted to given stress condition, GFP synthesis is induced and accumulated into the cytoplasm, leading to the increase of the cell's fluorescence. Flow cytometry detection is used in order to quantify the fluorescence at the single cell level. Obtained results are frequency histograms of fluorescenceintensity in the microbial population • RESULTS: The rpoS gene is a gene of the general stress response, mainly induced at the entrance to stationary phase (during a lack of glucose). The tracking of the GFP fluorescence linked to the activation / repression of the rpoS promoter gives good results. Indeed, there is appearance of a segregation at the level of the GFP content among the microbial population. The intensity of the segregation, as well as its time of appearance during the culture can be related to the bioreactor mixing efficiency. • CONCLUSION: prpoS::gfp strains can be used as biosensors of the heterogeneity of glucose encountered inside industrial reactors. • POTENTIAL APPLICATIONS & KEY BENEFITS: These strains could be used to validate a fed-batch regulation (addition of glucose) at the industrial level. | |
| Unité de bio-industries / Gembloux Agro Bio-Tech / Université de Liège | |
| Fond pour la recherche dans l'industrie et dans l'agriculture - FRIA | |
| Researchers ; Professionals ; Students | |
| http://hdl.handle.net/2268/79346 |
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