Metabolic profiling of bacteria with the application of polypyrrole-MOF SPME fibers and plasmonic nanostructured LDI-MS substrates

Abstract

Abstract Here we present application of innovative lab-made analytical devices such as plasmonic silver nanostructured substrates and polypyrrole-MOF solid-phase microextraction fibers for metabolic profiling of bacteria. For the first time, comprehensive metabolic profiling of both volatile and non-volatile low-molecular weight compounds in eight bacterial strains was carried out with utilization of lab-made devices. Profiles of low molecular weight metabolites were analyzed for similarities and differences using principal component analysis, hierarchical cluster analysis and random forest algorithm. The results showed clear differentiation between Gram positive (G+) and Gram negative (G−) species which were identified as distinct clusters according to their volatile metabolites. In case of non-volatile metabolites, differentiation between G+ and G− species and clustering for all eight species were observed for the chloroform fraction of the Bligh & Dyer extract, while methanolic fraction failed to recover specific ions in the profile. Furthermore, the results showed correlation between volatile and non-volatile metabolites, which suggests that lab-made devices presented in the current study might be complementary and therefore, useful for species differentiation and gaining insights into bacterial metabolic pathways.