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• W3020659518 startingPage "1336" @default.
• W3020659518 abstract "Modulating resource allocation in bacteria to redirect metabolic building blocks to the formation of recombinant proteins rather than biomass formation remains a grand challenge in biotechnology. Here, we present a novel approach for improved recombinant protein production (RPP) using Escherichia coli (E. coli) by decoupling recombinant protein synthesis from cell growth. We show that cell division and host mRNA transcription can be successfully inhibited by coexpression of a bacteriophage-derived E. coli RNA polymerase (RNAP) inhibitor peptide and that genes overtranscribed by the orthogonal T7 RNAP can finally account to >55% of cell dry mass (CDM). This RNAP inhibitor peptide binds the E. coli RNAP and therefore prevents σ-factor 70 mediated formation of transcriptional qualified open promoter complexes. Thereby, the transcription of σ-factor 70 driven host genes is inhibited, and metabolic resources can be exclusively utilized for synthesis of the protein of interest (POI). Here, we mimic the late phase of bacteriophage infection by coexpressing a phage-derived xenogeneic regulator that reprograms the host cell and thereby are able to significantly improve RPP under industrial relevant fed-batch process conditions at bioreactor scale. We have evaluated production of several different recombinant proteins at different scales (from microscale to 20 L fed-batch scale) and have been able to improve total and soluble proteins yields up to 3.4-fold in comparison to the reference expression system E. coli BL21(DE3). This novel approach for growth-decoupled RPP has profound implications for biotechnology and bioengineering and helps to establish more cost-effective and generic manufacturing processes for biologics and biomaterials." @default.
• W3020659518 created "2020-05-01" @default.
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• W3020659518 date "2020-04-23" @default.
• W3020659518 modified "2023-10-07" @default.
• W3020659518 title "Bacteriophage Inspired Growth-Decoupled Recombinant Protein Production in <i>Escherichia coli</i>" @default.
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• W3020659518 doi "https://doi.org/10.1021/acssynbio.0c00028" @default.
• W3020659518 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/32324989" @default.
• W3020659518 hasPublicationYear "2020" @default.
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