Recombinant protein production in an Escherichia coli reduced genome strain.
Recently, efforts have been made to improve the properties of Escherichia coli as a recombinant host by 'genomic surgery'-deleting large segments of the E. coli K12 MG1655 genome without scars. These excised segments included K-islands, which contain a high proportion of transposons, insertion sequences, cryptic phage, damaged, and unknown-function genes. The resulting multiple-deletion strain, designated E. coli MDS40, has a 14% (about 700 genes) smaller genome than the parent strain, E. coli MG1655. The multiple-deletion and parent E. coli strains were cultured in fed-batch fermenters to high cell densities on minimal medium to simulate industrial conditions for evaluating growth and recombinant protein production characteristics. Recombinant protein production and by-product levels were quantified at different controlled growth rates. These results indicate that the multiple-deletion strain's growth behavior and recombinant protein productivity closely matched the parent stain. Thus, the multiple-deletion strain E. coli MDS40 provides a suitable foundation for further genomic reduction.
Sharma
Shamik S
SS
Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, SC 29634, USA.
Blattner
Frederick R
FR
Harcum
Sarah W
SW
D049109:Cell Proliferation; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D017353:Gene Deletion; D018390:Gene Targeting / Q000379:methods; D024861:Genetic Enhancement / Q000379:methods*; D016680:Genome, Bacterial / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism*; D013045:Species Specificity
D029968:Escherichia coli Proteins; D011994:Recombinant Proteins