Functional genomics via metabolic footprinting: monitoring metabolite secretion by Escherichia coli tryptophan metabolism mutants using FT-IR and direct injection electrospray mass spectrometry

David Ian Ellis, Royston Goodacre, Naheed N. Kaderbhai, David Iain Broadhurst, Douglas B. Kell

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Abstract

We sought to test the hypothesis that mutant bacterial strains could be discriminated from each other on the basis of the metabolites they secrete into the medium (their ‘metabolic footprint’), using two methods of ‘global’ metabolite analysis (FT–IR and direct injection electrospray mass spectrometry). The biological system used was based on a published study of Escherichia coli tryptophan mutants that had been analysed and discriminated by Yanofsky and colleagues using transcriptome analysis. Wild-type strains supplemented with tryptophan or analogues could be discriminated from controls using FT–IR of 24 h broths, as could each of the mutant strains in both minimal and supplemented media. Direct injection electrospray mass spectrometry with unit mass resolution could also be used to discriminate the strains from each other, and had the advantage that the discrimination required the use of just two or three masses in each case. These were determined via a genetic algorithm. Both methods are rapid, reagentless, reproducible and cheap, and might beneficially be extended to the analysis of gene knockout libraries.
Original languageEnglish
Pages (from-to)376-391
Number of pages16
JournalComparative and Functional Genomics
Volume4
Issue number4
DOIs
Publication statusPublished - 18 Jul 2003

Keywords

  • metabolome
  • metabolomics
  • metabolic footprinting
  • mass spectromerty
  • FTIR spectroscopy
  • Escherichia coli
  • tryptophan
  • genetic algorithm

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