Web Release Date: June 19,
Monitoring of Recombinant Protein Production Using Bioluminescence in a Semiautomated Fermentation Process
and
Laboratoire d'Automatique et de Génie des Procédés (LAGEP), UMR CNRS 5007, Université Claude Bernard Lyon1 et ESCPE-Lyon, bât 308 G, 43 bd du 11 nov 1918, 69622 Villeurbanne Cedex, France; Unité de Génétique et de Microbiologie, UMR CNRS 5122 composante INSA, bât A. Lwoff, 10 rue Dubois, 69621 Villeurbanne Cedex, France; Plateau technique 5, Génomique Structurale, site Fernbach, Institut Pasteur, 25-28 rue du Dr Roux, 75724 Paris Cedex 15, France; and Department of Chemical Engineering, University of Delaware, Newark, Delaware 19716
Accepted for publication April 28, 2003.
Abstract:
On-line optimization of fermentation processes can be greatly aided by the availability of information on the physiological state of the cell. The goal of our "BioLux" research project was to design a recombinant cell capable of intracellular monitoring of product synthesis and to use it as part of an automated fermentation system. A recombinant plasmid was constructed containing an inducible promoter that controls the gene coding for a model protein and the genes necessary for bioluminescence. The cells were cultured in microfermenters equipped with an on-line turbidity sensor and a specially designed on-line light sensor capable of continuous measurement of bioluminescence. Initial studies were done under simple culture conditions, and a linear correlation between luminescence and protein production was obtained. Such specially designed recombinant bioluminescent cells can potentially be applied for model-based inference of intracellular product formation, as well as for optimization and control of recombinant fermentation processes.
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