Affiliation:
1. Institute of Biochemistry University of Greifswald Felix-Hausdorff-Strasse 4 17489 Greifswald Germany
2. Institute of Pharmacy University of Greifswald Friedrich-Ludwig-Jahn-Strasse 17 17489 Greifswald Germany
Abstract
AbstractBiofilms of pathogenic bacteria are responsible for persistent infections in humans, therefore investigations of biofilm formation and treatment strategies are required. The gram‐negative enterobacterium Escherichia (E.) coli is the most common pathogen causing chronic or recurring urinary tract infections. Metabolomics approaches targeted the bacterium to investigate specific metabolic patterns of biofilms and regulatory influences on biofilm formation. In this study, we aimed to investigate the metabolome of biofilms formed by the multidrug‐resistant extended‐spectrum beta‐lactamase‐producing (ESBL) E. coli PBIO729. For this purpose, a protocol for fast sampling of the macrocolony biofilms and efficient extraction of metabolites was optimized. Validation of an LC‐MS/MS method confirmed its usability for the analysis of nucleotides and other phosphorylated metabolites. A GC‐MS approach was used to monitor nutrient uptake from the medium in addition to the analysis of amino acid content and metabolites of glycolysis and TCA cycle in E. coli biofilms.