Metabolic footprinting of the anaerobic bacterium Fusobacterium varium using 1H NMR spectroscopy

Mol Biosyst. 2011 Jul;7(7):2220-7. doi: 10.1039/c1mb05105a. Epub 2011 May 6.

Abstract

Metabolic footprinting of the anaerobic bacterium Fusobacterium varium demonstrated the accumulation of six carboxylic acids as metabolic end-products and revealed specific growth requirements and utilization capabilities towards amino acids. Guided by (1)H NMR determinations of residual amino acids in spent medium, a modified chemically defined minimal medium (CDMM*) was developed by minimizing the amino acid composition while satisfying nutritional requirements to support abundant growth of F. varium. Quantitative determinations of carboxylate salts and residual substrates were readily performed by (1)H NMR analysis of lyophilized residues from CDMM* cultures without interference from initial medium components. Only small concentrations of alanine, arginine, glycine, isoleucine, leucine, methionine, proline and valine were required to support growth of F. varium, whereas larger quantities of aspartate, asparagine, cysteine, glutamine, glutamate, histidine, lysine, serine and threonine were utilized, most likely as energy sources. Both bacterial growth and the distribution of carboxylate end-products depended on the composition of the chemically defined medium. In cultures provided with glucose as the primary energy source, the accumulation of butyrate and lactate correlated with growth, consistent with the regeneration of reduced coenzyme formed by the oxidative steps of glucose catabolism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / metabolism
  • Amino Acids / pharmacology
  • Anaerobiosis / drug effects
  • Culture Media / pharmacology
  • Energy Metabolism / drug effects
  • Fusobacterium / drug effects
  • Fusobacterium / growth & development
  • Fusobacterium / metabolism*
  • Glucose / metabolism
  • Magnetic Resonance Spectroscopy
  • Metabolome / drug effects
  • Metabolomics / methods*
  • Protons*
  • Reference Standards
  • Reproducibility of Results

Substances

  • Amino Acids
  • Culture Media
  • Protons
  • Glucose