Off-line monitoring of bacterial stress response during recombinant protein production using an optical biosensor.
A surface plasmon resonance (SPR) biosensor was used to monitor the profiles of the heat-shock protein (DnaK) and the expression of a heterologous protein to map the dynamics of the cellular stress response in Escherichia coli. As expression system was used an E. coli strain overproducing human recombinant superoxide dismutase (rhSOD). Expression of DnaK showed complex patterns differing with strength of induction. The strong up-regulation of DnaK expression was observed in all cultivations which over-produced of rhSOD. Similar patterns were not observed in non-induced reference cultures. Differences in DnaK concentration profiles were correlated with induction strength. Presented data, carried out in shake flask and glucose limited fed-batch cultivation, show a good consistency with previously published transcriptional profiling results and provide complementary information to understand stress response related to overproduction of recombinant protein. The study also demonstrates the feasibility of using the SPR as a two channel protein array for monitoring of intracellular components.
Vostiar
Igor
I
Division of Biotechnology, Department of Physics and Measurement Technology, Linköping University, 581 83 Linköping, Sweden.
Tkac
Jan
J
Mandenius
Carl-Fredrik
CF
D019149:Bioreactors / Q000382:microbiology; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D049109:Cell Proliferation; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000032:analysis / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000378:metabolism*; D018869:Heat-Shock Response / Q000502:physiology*; D009862:Online Systems; D055095:Optics and Photonics / Q000295:instrumentation; D018384:Oxidative Stress / Q000502:physiology; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000235:genetics; D020349:Surface Plasmon Resonance / Q000295:instrumentation* / Q000379:methods
D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D011994:Recombinant Proteins; D013482:Superoxide Dismutase; C063842:dnaK protein, E coli