A Comparative Study of Global Stress Gene Regulation in Response to Overexpression of Recombinant Proteins in Escherichia coli

Role of theompTMutation in Stimulated Decrease in Colony-Forming Ability Due to Intracellular Protein Aggregate Formation inEscherichia coliStrain BL21

“…23,24) It is also known to be involved in the maturation of secreted proteins and in in vivo degradation of foreign proteins. 20,25,26) It is also known that ompT is expressed in E. coli producing recombinant proteins, 27,28) and that OmpT protease associates with intracellular protein aggregates, 29) indicating that it plays a central role in E. coli protein quality control. Mutants lacking this enzyme are constructed and are used as expression hosts for recombinant proteins.…”

Recombinant protein production in an Escherichia coli reduced genome strain

“…Although E. coli is a well studied and a commonly used recombinant host, the behavior of E. coli can still be unpredictable at times, especially while expressing recombinant protein (Andersson et al, 1996;Baneyx, 1999;Baneyx and Mujacic, 2004;Gill et al, 2000;Oh and Liao, 2000;Sanden et al, 2003;Swartz, 1996). One of the overall goals of the multiple deletion strain project is to improve the growth/yield properties and robustness of E. coli as a recombinant host by eliminating potentially nonessential genes.…”

Increasing Recombinant Protein Production in E. coli by an Alternative Method to Reduce Acetate

“…The production of heterologous proteins to high titres concurs mostly with the initiation of a stress response and/or metabolic burden, both associated with the use of multicopy plasmids, resulting in misfolding and degradation of the heterologous protein and formation of inclusion bodies (Noack et al, 1981;Parsell & Sauer, 1989;Bentley et al, 1990;Gill et al, 2000;Hoffmann & Rinas, 2004;Ventura & Villaverde, 2006). ii.…”

“…However, problems such as metabolic burden, segregational instability, misfolding and proteolytic breakdown or aggregation in inclusion bodies, and difficulties in controlling gene expression are usually associated with multi-copy plasmids and the use of strong promoters (Noack et al, 1981;Parsell & Sauer, 1989;Bentley et al, 1990;Dong et al, 1995;Kurland & Dong, 1996;Gill et al, 2000;Hoffmann & Rinas, 2004;Ventura & Villaverde, 2006). Most engineering strategies to tackle these problems focus on prevention of misfolding, neutralisation of increased protease activity or stress response (Chou, 2007).…”