Modelling the stability of the pBR322 plasmid derivative pBB210 in Escherichia coli TG1 under non-selective and selective conditions

A mathematical model has been developed to describe the stability behaviour of the pBR322 plasmid derivative pBB210 with the β-lactamase gene and the human interferon-α1 gene in Escherichia coli TG1 under non-selective, selective and modified selective conditions in a chemostat. The model was formulated on the basis of experimental investigations. It includes the interaction between β-lactam antibiotics (ampicillin and sulbactam) and cells (with and without plasmids), in particular the correlation between the growth rate of plasmid-free cells and ampicillin concentration in the medium; ampicillin transport into the periplasm of the plasmid-bearing cells; ampicillin degradation in the periplasm by by plasmid-encoded β-lactamase and the inhibition of the latter by sulbactam. The results obtained by the simulation of chemostat cultivations under various conditions and by steady state analyses are closely related to the results of experiments. Under non-selective conditions, the fraction of plasmid-bearing cells was approaching zero. Under selective and modified selective conditions, a coexistence between plasmid-free and plasmid-bearing cells was reached at steady state. Under these conditions, the steady state fraction of plasmid-bearing cells was proportional to the ampicillin concentration in the feed and inversely proportional to the cell concentration in the chemostat. During high-density cultivation, a large amount of ampicillin is necessary to suppress plasmid-free cells. Even small concentrations of the β-lactamase inhibitor sulbactam in the feed increased the steady state fraction of plasmid-bearing cells (from 17.2% to 99.6% at sulbactam-Na concentrations of 0 to 5 mg/l).