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Related Experiment Videos

Rational medium design for Bordetella pertussis: basic metabolism.

M Thalen1, J van den IJssel, W Jiskoot

  • 1National Institute of Public Health and the Environment (RIVM), Laboratory for Product and Process Development, Bilthoven, The Netherlands. marcel.thalen@rivm.nl

Journal of Biotechnology
|December 20, 1999
PubMed
Summary
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Optimizing Bordetella pertussis culture media by understanding its metabolism prevents ammonium accumulation and metabolite excretion. Using lactate and glutamate as substrates significantly improves biomass yield in fed-batch cultures.

Area of Science:

  • Microbiology
  • Biotechnology
  • Metabolic Engineering

Background:

  • Ammonium accumulation limits Bordetella pertussis cell density in fed-batch cultures due to substrate imbalances.
  • B. pertussis exhibits partially dysfunctional glycolysis and citric-acid cycle, impacting its metabolism.

Purpose of the Study:

  • To map Bordetella pertussis catabolic and anabolic capabilities.
  • To design a culture medium that prevents ammonium accumulation and ensures complete substrate metabolism.
  • To enhance biomass yield in B. pertussis cultures.

Main Methods:

  • Utilized nuclear magnetic resonance (NMR) and electron microscopy to analyze B. pertussis metabolism.
  • Investigated various carbon sources, including glutamate, lactate, and combinations thereof.

Related Experiment Videos

  • Assessed metabolite excretion and poly-hydroxy-butyrate (PHB) globule formation.

    • Main Results:

    • Adding various carbon sources to glutamate-containing media avoided ammonium accumulation but led to acetate and beta-hydroxy-butyrate excretion.
    • Poly-hydroxy-butyrate (PHB) globules were observed, coinciding with beta-hydroxy-butyrate and acetoacetate formation.
    • Using lactate with glutamate as substrates prevented metabolite excretion and ammonium accumulation, achieving a high biomass yield (8.8 g C-mol⁻¹).

    Conclusions:

    • A novel medium using lactate and glutamate effectively supports B. pertussis growth without ammonium accumulation or metabolite excretion.
    • This optimized medium significantly improves biomass yield compared to existing literature values.
    • Further optimization for pertussis toxin production is detailed in a separate study.