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    Research study on phage-inspired growth decoupled recombinant protein production in Escherichia coli

    news-releasesenGenes Biotech GmbH
    May 28th 2020

    Vienna, Austria: – Scientists from recombinant proteins specialist CDMO enGenes Biotech GmbH (enGenes) have collaborated with researchers from Vienna’s University of Natural Resources and Life Sciences (BOKU) on an important study of bacteriophage Inspired Growth-Decoupled Recombinant Protein Production in Escherichia coli.

    The research study was authored by four enGenes scientists, Patrick Stargardt, Lukas Feuchtenhofer, and Juergen Mairhofer in partnership with Monika Cserjan-Puschmann and Prof. Gerald Striedner from BOKU.

    Decoupling synthesis from cell growth

    The article, accepted for publication in the American Chemical Society (ACS) Synthetic Biology journal, presents a novel approach for improved recombinant protein production (RPP) using E. coli by decoupling recombinant protein synthesis from cell growth and outlines the capabilities of the enGenes-eXpress technology within a peer-reviewed format.

    The authors show that cell division and host mRNA transcription can be successfully inhibited by co-expression of a bacteriophage-derived E. coli RNA polymerase (RNAP) inhibitor peptide and that genes over-transcribed by the orthogonal T7 RNAP can finally account for more than 55 per cent of cell dry mass (CDM).

    Beating the biomass challenge

    This helps to address a principal challenge in biotechnology; how to modulate resource allocation in bacteria to redirect metabolic building blocks to the formation of recombinant proteins rather than biomass.

    In the study, the 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).

    Improving RPP at bioreactor scale

    Lead author Patrick Stargardt comments: “Here, we mimic the late phase of bacteriophage infection by co-expressing 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 and even at 2500 L 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),” Mr. Stargardt explains.

    “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,” he concludes.

    About enGenes Biotech

    enGenes Biotech GmbH (enGenes) is a contract research, development and manufacturing company that provides leading edge technologies and production services focused on recombinant proteins in bacteria. The company’s mission is to provide cost-effective and scalable production of recombinant proteins at a fraction of current cost, allied to a vision of developing a world-class portfolio of cutting-edge protein production technologies, relevant to a broad spectrum of application fields.

    enGenes has developed advanced technologies to drive more cost-effective recombinant protein production processes, including its proprietary enGenes-eXpress™ E. coli platform that achieves outstanding yields of soluble and active recombinant protein. enGenes-eXpress™ has been successfully applied for the manufacturing of enzymes and biopharmaceutical products that failed to give economically feasible yields in a conventional expression host.

    enGenes Biotech offers development and manufacturing services tailored around the needs of pharmaceutical and industrial biotech companies. The services include expression strain and vector development, fermentation process development and optimization, downstream process development, production of purified protein, technology transfer and scale-up support with technology out-licensing and co-development opportunities.


    To access full paper click on Bacteriophage Inspired Growth-Decoupled Recombinant Protein Production in Escherichia coli

    Research study on phage-inspired growth decoupled recombinant protein production in Escherichia coli

    Phage-assisted RPP process described in enGenes/BOKU research study

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