High cell density culture of recombinant E. coli in the miniaturized bubble columns

Miniaturized bubble columns (MBCs) can provide mass transfer characteristics similar to stirred tank bioreactors. In this study, a new application was developed for MBCs to investigate the effect of feeding strategy and medium type on the fed-batch culture of recombinant E. coli. The results showed that the exponential feeding strategy and defined M9 medium were more suitable to achieve the high cell density culture (HCDC). The maximum obtained cell concentration in exponential feeding strategy in the defined medium without induction, was at OD₆₀₀ of 169, while glucose concentration was maintained under 2 g/L. To the best of our knowledge, this cell concentration cannot be achieved in lab or pilot scale bubble columns. At the end of the process, adverse effect of the metabolic burden due to induction and mass transfer limitations decreased the obtained final cell concentration to OD₆₀₀ of 116. Finally, a comparison of the results for fed-batch culture in the stirred tank bioreactor with those of the MBCs showed that their lower cell concentrations were due to the hydrodynamics limitations of MBCs. Yet, it was found that the MBCs are efficient tools in development of feeding strategies and evaluation of medium components for HCDC of recombinant E. coli.