It was of major interest to know whether BRP-mediated secretion of recombinant proteins could be adopted in a stable chemostat and if so, what kind of interplay would be seen between activity of growth-phase regulated promoters and the productivity of a chemostat process. To make this continuous process even more sustainable, alternative plasmid selection mechanisms were tested with the aim of avoiding antibiotic-based plasmid selection. Through the use of auxotrophy complementation, it was possible to maintain a previously unstable plasmid, for extended periods of continuous cultivation without any antibiotic-based selection pressure and show extracellular expression of a recombinant enzyme, thus providing an interesting new model.
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