Simulation and prediction of protein production in fed‐batch E. coli cultures: An engineering approach

Abstract

An overall model describing the dynamic behavior of fed-batch E. coli processes for protein production has been built, calibrated and validated. Using a macroscopic approach, the model consists of three interconnected blocks allowing simulation of biomass, inducer and protein concentration profiles with time. The model incorporates calculation of the extra and intracellular inducer concentration, as well as repressor-inducer dynamics leading to a successful prediction of the product concentration. The parameters of the model were estimated using experimental data of a rhamnulose-1-phosphate aldolase-producer strain, grown under a wide range of experimental conditions. After validation, the model has successfully predicted the behavior of different strains producing two different proteins: fructose-6-phosphate aldolase and ω-transaminase. In summary, the presented approach represents a powerful tool for E. coli production process simulation and control.

Synthesis of a precursor of D-fagomine by immobilized fructose-6-phosphate aldolase

“…Recombinant FSA A129S was produced and purified according to the literature with an activity of 15.0 U mg -1 FSA (S1 Fig) [38]. N- (3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDAC), α-glycerophosphate dehydrogenase-triosephosphate isomerase (GPD-TPI), D-fructose-6-phosphate (F6P) and 3-[(benzyloxycarbonyl)amino]propionaldehyde (β-CHO) were purchased from Sigma Aldrich (St. Louis, USA).…”