Direct and indirect use of GFP whole cell biosensors for the assessment of bioprocess performances: Design of milliliter scale-down bioreactors

Authors

  • Alison Brognaux,

    1. Université de Liège, Gembloux Agro-Bio Tech, Unité de Bio-industries/CWBI, Passage des Déportés 2, B-5030 Gembloux, Belgium
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  • Philippe Thonart,

    1. Université de Liège, Gembloux Agro-Bio Tech, Unité de Bio-industries/CWBI, Passage des Déportés 2, B-5030 Gembloux, Belgium
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  • Frank Delvigne,

    Corresponding author
    1. Université de Liège, Gembloux Agro-Bio Tech, Unité de Bio-industries/CWBI, Passage des Déportés 2, B-5030 Gembloux, Belgium
    • Université de Liège, Gembloux Agro-Bio Tech, Unité de Bio-industries/CWBI, Passage des Déportés 2, B-5030 Gembloux, Belgium

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  • Peter Neubauer,

    1. Dept. of Biotechnology, Technische Universität Berlin, Laboratory of Bioprocess Engineering, Ackerstr. 71-76, D-13355 Berlin, Germany
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  • Jean-Claude Twizere,

    1. Université de Liège, Gembloux Agro-Bio Tech, Unité de biologie cellulaire et moléculaire, Avenue Maréchal Juin 13, B-5030 Gembloux, Belgium
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  • Frédéric Francis,

    1. Université de Liège, Gembloux Agro-Bio Tech, Unité d'entomologie fonctionnelle et évolutive, Passage des Déportés 2, B-5030 Gembloux, Belgium
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  • Nathalie Gorret

    1. Université de Toulouse; INSA, UPS, INP, LISBP, INRA, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, 135 Avenue de Rangueil, F-31077 Toulouse, France
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Abstract

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. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2013

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