NMR-based metabolomics of <i>Burkholderia pseudomallei</i> biofilms and extracellular polymeric substance cultured in LB and MVBM media

<i>Burkholderia pseudomallei</i> biofilms are resistant to antibiotics and immune responses, leading to persistent infections. This study aimed to investigate the metabolic profiles of <i>B. pseudomallei</i> in biofilms and the extracellular polymeric substances (EPS) produced during grown in LB or MVBM medium using Nuclear Magnetic Resonance (NMR) spectroscopy to identify key metabolites. The results revealed similar biofilm metabolites in both media. However, betaine was detected in LB, but not in the case of MVBM. Acetate was significantly higher in MVBM compared to that of LB. Pathway analysis revealed that betaine-producing <i>B. pseudomallei</i> biofilm in LB was associated with metabolism of glycine, serine, and threonine, while acetate in MVBM was associated with metabolism of taurine and hypotaurine, phosphonate and phosphinate, and glycolysis/gluconeogenesis. The NMR analysis of EPS disclosed shared metabolites including dimethylsulfide, 1-methyluric acid and oxypurinol. This study provides the first extensive investigation into <i>B. pseudomallei</i> biofilm and EPS metabolites, identifying pathways that offer potential targets for combating <i>B. pseudomallei</i> biofilm-associated infections.