Quantitative metabolomics of the thermophilic methylotroph Bacillus methanolicus

Marc Carnicer, Gilles Vieira, Trygve Brautaset, Jean Charles Portais, Stephanie Heux

Research output: Indexed journal article Articlepeer-review

18 Citations (Scopus)

Abstract

Background: The gram-positive bacterium Bacillus methanolicus MGA3 is a promising candidate for methanol-based biotechnologies. Accurate determination of intracellular metabolites is crucial for engineering this bacteria into an efficient microbial cell factory. Due to the diversity of chemical and cell properties, an experimental protocol validated on B. methanolicus is needed. Here a systematic evaluation of different techniques for establishing a reliable basis for metabolome investigations is presented. Results: Metabolome analysis was focused on metabolites closely linked with B. methanolicus central methanol metabolism. As an alternative to cold solvent based procedures, a solvent-free quenching strategy using stainless steel beads cooled to 20 °C was assessed. The precision, the consistency of the measurements, and the extent of metabolite leakage from quenched cells were evaluated in procedures with and without cell separation. The most accurate and reliable performance was provided by the method without cell separation, as significant metabolite leakage occurred in the procedures based on fast filtration. As a biological test case, the best protocol was used to assess the metabolome of B. methanolicus grown in chemostat on methanol at two different growth rates and its validity was demonstrated. Conclusion: The presented protocol is a first and helpful step towards developing reliable metabolomics data for thermophilic methylotroph B. methanolicus. This will definitely help for designing an efficient methylotrophic cell factory.

Original languageEnglish
Article number92
JournalMicrobial Cell Factories
Volume15
Issue number1
DOIs
Publication statusPublished - 1 Jun 2016
Externally publishedYes

Keywords

  • Bacillus methanolicus
  • Methanol
  • Quantitative metabolomics
  • Quenching

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