Fast Sampling of the Cellular Metabolome.

Area of Science:

• Microbiology
• Metabolomics
• Biochemistry

Background:

• Metabolomics aims to capture cellular metabolite profiles.
• Accurate microbial metabolomics requires rapid sampling, quenching, and extraction to prevent metabolite changes.
• Extracellular metabolites can interfere with intracellular metabolite quantification.

Purpose of the Study:

• To review and present methods for microbial metabolomics.
• To provide protocols for sampling, quenching, and extraction.
• To address challenges in quantifying microbial endo-metabolomes, especially for prokaryotes.

Main Methods:

• Overview of literature-described sampling, quenching, and extraction techniques.
• Detailed protocols for rapid sampling, quenching, and extraction.
• Measurement of metabolites in total broth, washed cells, and supernatant for differential analysis.

Main Results:

• Established methods for rapid microbial sample processing.
• Protocols applicable to both eukaryotic and prokaryotic microorganisms.
• A differential method is proposed for accurate prokaryotic endo-metabolome quantification.

Conclusions:

• Effective microbial metabolomics relies on optimized sampling, quenching, and extraction.
• The differential method is essential for accurate prokaryotic metabolomics.
• The presented protocols facilitate quantitative metabolomics in diverse microorganisms.