Fatty acid methyl ester (FAME) profiling identifies carbapenemase-producing Klebsiella pneumoniae belonging to clonal complex 258

Carbapenem-resistant Klebsiella pneumoniae (CRKP) is one of the most extensively antibiotic-resistant pathogens encountered in the clinical setting today. A few studies to-date suggest that CRKP and carbapenem-susceptible K. pneumoniae (CSKP) differ from one another not only with respect to their underlying genetics, but also their transcriptomic and metabolomic fingerprints. Within this context, we characterize the fatty acid methyl ester (FAME) profiles of these pathogens in vitro. Specifically, we evaluated the FAME profiles of six Klebsiella pneumoniae carbapenemase (KPC)-producing isolates belonging to the CC258 lineage (KPC+/258+), six KPC-producing isolates belonging to non-CC258 lineages (KPC+/258−), and six non-KPC-producing isolates belonging to non-CC258 lineages (KPC−/258−). We utilized a single-step sample preparation method to simultaneously lyse bacterial cells and transesterify the lipid fraction, and identified 14 unique FAMEs using gas chromatography-mass spectrometry. The machine learning algorithm Random Forest identified four FAMEs that were highly discriminatory between CC258 and non-CC258 isolates (9(Z)-octadecenoate, 2-phenylacetate, pentadecanoate, and hexadecanoate), of which three were also significantly different in relative abundance between these two groups. These findings suggest that distinct differences exist between CC258 and non-CC258 K. pneumoniae isolates with respect to the metabolism of both fatty acids and amino acids, a hypothesis that is supported by previously-acquired transcriptomic data.