title: High-performance recombinant protein production with Escherichia coli in continuously operated cascades of stirred-tank reactors.
abstract: The microbial expression of intracellular, recombinant proteins in continuous bioprocesses suffers from low product concentrations. Hence, a process for the intracellular production of photoactivatable mCherry with Escherichia coli in a continuously operated cascade of two stirred-tank reactors was established to separate biomass formation (first reactor) and protein expression (second reactor) spatially. Cascades of miniaturized stirred-tank reactors were implemented, which enable the 24-fold parallel characterization of cascade processes and the direct scale-up of results to the liter scale. With PAmCherry concentrations of 1.15 g L-1cascades of stirred-tank reactors improved the process performance significantly compared to production processes in chemostats. In addition, an optimized fed-batch process was outperformed regarding space-time yield (149 mg L-1h-1). This study implicates continuous cascade processes to be a promising alternative to fed-batch processes for microbial protein production and demonstrates that miniaturized stirred-tank reactors can reduce the timeline and costs for cascade process characterization.
authors.affiliation: Institute of Biochemical Engineering, Technical University of Munich, Boltzmannstr. 15, 85748, Garching, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Schmideder
authors.affiliation: Institute of Biochemical Engineering, Technical University of Munich, Boltzmannstr. 15, 85748, Garching, Germany. d.weuster-botz@lrz.tum.de.
authors.forename: Dirk
authors.initials: D
authors.lastname: Weuster-Botz
mesh_terms: D061249:Batch Cell Culture Techniques; D018533:Biomass; D019149:Bioreactors; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis*
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins
keywords: Cascades of stirred-tank reactors; Continuous culture; Escherichia coli; High-throughput bioprocess design; Protein expression