Back to Symposium home page 12th International Symposium on Bioluminescence & Chemiluminescence

Symposium abstracts:

Use of bioluminescence for on-line estimation of the production of recombinant proteins

Trezzani, I.1,4, Nadri, M.1, Hammouri, H.1, Lieto, J.1, Dorel, C.2, Lejeune, P.2, Dhurjati, P.3, Bellalou, J.4, Longin, R.4

1. Laboratoire d’Automatique et de Génie des Procédés (LAGEP), UMR CNRS 5007, Université Claude Bernard et ESCPE Lyon, bât 308G, 43 bd du 11 novembre 1918, 69622 Villeurbanne Cedex, France

2. Laboratoire de Génétique Moléculaire des Micro-organismes et des Interactions Cellulaires, INSA Lyon, 20 av. Albert Einstein, 69621 Villeurbanne Cedex, France

3. Dept of Chemical Engineering, Univ. Delaware, Newark, Delaware 19716, USA

4. Laboratoire des Fermentations, Institut Pasteur, 25-28 rue du Dr Roux, 75724 Paris Cedex 15, France.

Email : rlongin@pasteur.fr

Optimization of product formation in a fermentation process is greatly facilitated by on-line product measurements. Unfortunately, most products require time consuming off-line assays. One solution is to "estimate" the product levels from the available fermentation measurements. We used bioluminescence as a macroscopic indicator for the estimation of a microscopic parameter, intracellular product formation. A recombinant plasmid was constructed containing the PBAD promoter induced by arabinose, which controls the lacZ gene coding for b -galactosidase as a model protein, and the luxCDABE genes coding for bioluminescence. Bioluminescence and turbidity were measured on-line by specially designed sensors.... Correlations between bioluminescence and enzyme production were obtained in a minimal medium with glycerol and in a complex medium with higher cell density cultures. The objective was to infer at any time the amount of product produced in the reactor by the cells, using a model that relates product to the on-line measurements. This initial model opens up the possibility for optimization and control of the fermentation process.

This is a preprint of an article accepted for publication in Luminescence: Copyright 2001 John
Wiley & Sons, Ltd (Wiley website)