title: Tunable recombinant protein expression with E. coli in a mixed-feed environment.
abstract: Controlling the recombinant protein production rate in Escherichia coli is of utmost importance to ensure product quality and quantity. Up to now, only the genetic construct, introduced into E. coli, and the specific growth rate of the culture were used to influence and stir the productivity. However, bioprocess technological means to control or even tune the productivity of E. coli are scarce. Here, we present a novel method for the process-technological control over the recombinant protein expression rate in E. coli. A mixed-feed fed-batch bioprocess based on the araBAD promoter expression system using both D-glucose and L-arabinose as assimilable C-sources was designed. Using the model product green fluorescent protein, we show that the specific product formation rate can be efficiently tuned even on the cellular level only via the uptake rate of L-arabinose. This novel approach introduces an additional degree of freedom for the design of recombinant bioprocesses with E. coli. We anticipate that the presented method will result in significant quality and robustness improvement as well as cost and process time reduction for recombinant bacterial bioprocesses in the future.
authors: Institute of Biochemical Engineering, Vienna University of Technology, Gumpendorfer strasse 1A/166-4, Vienna 1060, Austria.
authors: Patrick
authors: P
authors: Sagmeister
authors: Clemens
authors: C
authors: Schimek
authors: Andrea
authors: A
authors: Meitz
authors: Christoph
authors: C
authors: Herwig
authors: Oliver
authors: O
authors: Spadiut
mesh_terms: D001089:Arabinose / Q000378:metabolism; D001431:Bacteriological Techniques / Q000379:methods*; D001709:Biotechnology / Q000379:methods; D002244:Carbon / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005947:Glucose / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D002244:Carbon; D001089:Arabinose; D005947:Glucose