Characterization of stress and protein turnover from protein overexpression in fed-batch E. coli cultures.
A structured kinetic model that accounts for proteolytic degradation due to recombinant protein overexpression is introduced and its performance evaluated by comparison with previously reported fed-batch experimental data. This mathematical model contains an additional pool for a generic key precursor (in our case phenylalanine), an improved IPTG transport term, a phenylalanine transport term, and a variable protein turnover expression that accounts for proteolytic activity. The model predictions concerning proteolytic activity, glucose level, and cell growth are in very good agreement with an amino acid depletion hypothesis. Cultures exposed to greater stress showed higher and/or longer proteolysis, whereas less overall proteolytic activity was observed when the effect of induction was somewhat ameliorated.
Ramírez
D M
DM
Center for Agricultural Biotechnology, University of Maryland Biotechnology Institute, College Park 20742, USA.
Bentley
W E
WE
D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D005285:Fermentation; D007700:Kinetics; D008433:Mathematics; D008961:Models, Structural; D011994:Recombinant Proteins / Q000378:metabolism*
D011994:Recombinant Proteins; D015500:Chloramphenicol O-Acetyltransferase