Reference : Direct and indirect use of GFP whole cell biosensors for the assessment of bioprocess pe...
Scientific journals : Article
Life sciences : Biotechnology
http://hdl.handle.net/2268/134258
Direct and indirect use of GFP whole cell biosensors for the assessment of bioprocess performances: design of milliliter scale-down bioreactors
English
Brognaux, Alison mailto [Université de Liège - ULg > Département des sciences de la vie > Biochimie et microbiologie industrielles >]
Neubauer, Peter []
Twizere, Jean-Claude mailto [Université de Liège - ULg > Chimie et bio-industries > Biologie cell. et moléc. >]
Francis, Frédéric mailto [Université de Liège - ULg > Sciences agronomiques > Entomologie fonctionnelle et évolutive >]
Gorret, Nathalie []
Thonart, Philippe mailto [Université de Liège - ULg > Département des sciences de la vie > Biochimie et microbiologie industrielles >]
Delvigne, Frank mailto [Université de Liège - ULg > Chimie et bio-industries > Bio-industries >]
2013
Biotechnology Progress
American Chemical Society
29
1
48-59
Yes (verified by ORBi)
International
8756-7938
1520-6033
[en] microbial stress ; scale-up ; scale-down ; fluorescent reporter ; protein leakage
[en] 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.
Fonds pour la formation à la Recherche dans l'Industrie et dans l'Agriculture (Communauté française de Belgique) - FRIA
Researchers ; Professionals ; Students
http://hdl.handle.net/2268/134258

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