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Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor
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NMR bioreactor development for live in-situ microbial functional analysis.

Paul D Majors1, Jeffrey S McLean, Johannes C M Scholten

  • 1Biological Sciences Division, Pacific Northwest National Laboratory, 3335 Q Avenue, MSIN: K8-98, Richland, WA 99352, USA. paul.majors@pnl.gov

Journal of Magnetic Resonance (San Diego, Calif. : 1997)
|March 4, 2008
PubMed
Summary

This study developed live, in-situ metabolomics for microbial cultures, revealing Eubacterium aggregans produces significant lactate, a novel finding for biofuel research.

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Area of Science:

  • Microbiology
  • Metabolomics
  • Bioengineering

Background:

  • Prokaryotic metabolomics is crucial for understanding microbial functions, particularly in industrial applications like biofuel production.
  • Existing methods often lack the capability for real-time, in-situ analysis of microbial metabolism under controlled conditions.
  • Eubacterium aggregans is an anaerobic bacterium relevant to biofuel production, but its metabolic pathways require further elucidation.

Purpose of the Study:

  • To develop and validate a live, in-situ metabolomics platform for prokaryotic cultures.
  • To investigate the metabolic profile of Eubacterium aggregans under controlled batch and continuous culture conditions.
  • To identify novel metabolic products or pathways in E. aggregans relevant to biofuel production.

Main Methods:

  • Development of a radiofrequency-transparent bioreactor integrated with a wide-bore nuclear magnetic resonance (NMR) imaging spectrometer and bioreactor controller.
  • Application of water-suppressed 1H NMR spectroscopy for real-time monitoring of substrate utilization and byproduct excretion.
  • Cultivation of Eubacterium aggregans under controlled batch and continuous growth conditions.

Main Results:

  • Successful implementation of a live, in-situ metabolomics capability for monitoring microbial cultures.
  • Quantification of glucose and fructose utilization and excretion of metabolites like short-chain organic acids and ethanol by E. aggregans.
  • Discovery that E. aggregans produces lactate as a significant end product, a previously unreported metabolic output.

Conclusions:

  • The developed live, in-situ metabolomics platform provides real-time insights into microbial metabolism.
  • The identification of lactate production in E. aggregans expands the understanding of its metabolic capabilities.
  • This approach complements functional genomics and systems biology, offering a powerful tool for microbial research and biotechnology.