The proteases of Escherichia coli

Thus far, only a handful of the around eighty proteases found in E. coli have been well characterized, and it is generally assumed that most of the uncharacterized proteases are of less importance.

However, my laboratory has recently shown that many of these ‘less important’ proteases can interfere with the production of recombinant proteins. This suggests that many proteases have not yet been studied under for them relevant conditions.

Therefore, I propose to use a combination of advanced bacterial genetics, microbial physiology, transcriptomics and proteomics to characterize the totality of the proteases of E. coli under a variety of naturally occurring stress conditions and recombinant protein production stress by -i- analysing the regulation of their genes, -ii- determining how they contribute to growth and stress relief, and -iii- identifying their interacting partners and substrates.

Since E. coli has successfully been used before to further our understanding of protein homeostasis in other pro- and eukaryotes, it is envisaged that the knowledge generated will be widely applicable. It will also enable to further develop E. coli as cost- and time-effective recombinant protein production host.

The aims of the project reflect the long-standing goals of the laboratory: -i- to provide a global picture of how protein homeostasis in bacteria is maintained, and -ii- to improve E. coli as a recombinant protein production host.