Biotechnologie, Agronomie, Société et Environnement (Jan 2010)
Dynamic analysis of microbial behavior face to environmental heterogeneities encountered in large-scale bioreactors [abstract]
Abstract
Heterogeneities caused by deficient mixing in large-scale bioreactors have been identified and described in literature. These heterogeneities affect physiological changes of microorganisms through its passage in different zones of concentrations. Consequently the differences in terms of productivities, qualities and/or yields of products of interest have been observed during scaling-up from laboratory to larger scales. For this reason, large-scale process improvement depends on the understanding of dynamic interactions between microbial responses and physical phenomena inside bioreactors. The dynamic responses of microorganisms are used as a tool for gaining insight into the fundamentals of microbial changes under a mixing-well controlled environment. Our research group has not only applied scale-down methodology to study the microbial responses at molecular, microscopic and macroscopic levels of observation, but also has used innovative strains and process engineering tools to evaluate fast dynamic responses of microorganism at time scales from seconds to minutes. This presentation focuses on the application of rational strategies in order to characterize distributed relaxation times of microorganisms which are considered to be constant at all fermentation scales. Experiments were conducted with reporter bioluminescent strains of Escherichia coli in which the luxCDABE operon was fused to promoters responding to different selected environmental stresses (dissolved oxygen, pH, temperature and substrate, etc.). Such strains allow real-time recording of the expression of genes involved in stress responses. Kinetic analyses of biomass, extracellular metabolites, inlet/exhaust gas, were carried out in order to determine mass balances and biological kinetic parameters. We will present our approach and the results concerning continuous culture of E. coli DPD2417 (nirB::luxCDABE) to monitor the microbial responses to oxygen limitation.
