recombinant protein production in e.coli

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Recombinant protein production in E.coli. Molecular and process strategies. Plasmid maintenance and copy number. Plasmid stability : Segregational stability Structural stability Host cell mutation. Improving plasmid stability. Programmed cell death upon loss of plasmid - PowerPoint PPT Presentation

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  • Recombinant protein production in E.coliMolecular and process strategies

  • Plasmid maintenance and copy numberPlasmid stability :

    Segregational stability

    Structural stability

    Host cell mutation

  • Improving plasmid stabilityProgrammed cell death upon loss of plasmidIntegrate gene of interest in chromosome gene copy number ?Combination of selection pressure and growth rate control by limiting the carbon source in the medium

  • Expression vector design consideration

  • High Cell Density cultivationDevelopment of growth mediaAcetate formationOxygen concentrationMixing patterns in large scale fermentersDissolved carbondioxide

  • Above the following concentrations E.coli is inhibited.Glucose 50 g/lAmmonia 3 g/lPhosphorus 10 g/lMagnesium 8.7 g/lZinc -0.038 g/lIron 1.15 g/lHence fed batch is the most preferable method

  • General methods applied to reduce acetate formation in E. coli

  • General methods applied to reduce acetate formation in E. coli

  • Oxygen concentrationPure oxygen can be sparged. But expensiveAnaerobic conditions lead to acetate formation MixingCells close to the injection port are exposed to high concentrated nutrients.

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