A new extant respirometric assay to estimate intrinsic growth parameters applied to study plasmid metabolic burden

Authors

  • Jose Seoane,

    1. Department of Environmental Engineering, Technical University of Denmark, Lyngby DK 2800, Denmark; telephone: +45-45-25-1600; fax: +45-45-93-28-50
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  • Gürkan Sin,

    1. Department of Chemical and Biochemical Engineering, Technical University of Denmark, Lyngby, Denmark
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  • Laurent Lardon,

    1. Laboratory of Environmental Biotechnology, Institute Nationale de la Recherche Agronomique, Narbonne, France
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  • Krist V. Gernaey,

    1. Department of Chemical and Biochemical Engineering, Technical University of Denmark, Lyngby, Denmark
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  • Barth F. Smets

    Corresponding author
    1. Department of Environmental Engineering, Technical University of Denmark, Lyngby DK 2800, Denmark; telephone: +45-45-25-1600; fax: +45-45-93-28-50
    • Department of Environmental Engineering, Technical University of Denmark, Lyngby DK 2800, Denmark; telephone: +45-45-25-1600; fax: +45-45-93-28-50.
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Abstract

Start-up phenomena in microbial biokinetic assays are not captured by the most commonly used growth-related equations. In this study we propose a new respirometric experimental design to estimate intrinsic growth parameters that allow us to avoid these limitations without data omission, separate mathematical treatment, or wake-up pulses prior to the analysis. Identifiability and sensitivity analysis were performed to confirm the robustness of the new approach for obtaining unique and accurate estimates of growth kinetic parameters. The new experimental design was applied to establish the metabolic burden caused by the carriage of a pWW0 TOL plasmid in the model organism Pseudomonas putida KT2440. The metabolic burden associated was manifested as a reduction in the yield and the specific growth rate of the host, with both plasmid maintenance and the over-expression of recombinant proteins from the plasmid contributing equally to the overall effect. Biotechnol. Bioeng. 2010;105: 141–149. © 2009 Wiley Periodicals, Inc.

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