Recombinant protein production in an Escherichia coli reduced genome strain |
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Authors: | Sharma Shamik S Blattner Frederick R Harcum Sarah W |
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Institution: | Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, SC 29634, USA. |
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Abstract: | 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. |
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