Summary
Strategy of managing antibiotic‐resistant Vibrio alginolyticus, a bacterial pathogen that threatens human health and animal farming, is not available due to the lack of knowledge about the underlying mechanism of antibiotic resistance. Here, we showed that gentamicin‐resistant V. alginolyticus (VA‐RGEN) has four mutations on metabolism and one mutation on a two‐component system by whole‐genome and PCR‐based sequencing, indicating the metabolic shift in VA‐RGEN. Thus, metabolic profile was investigated by GC–MS based metabolomics. Glucose was identified as a crucial biomarker, whose abundance was decreased in VA‐RGEN. Further analysis with iPath, and gene expression and enzyme activity of the pyruvate cycle (the P cycle) demonstrated a global depressed metabolic pathway network in VA‐RGEN. Consistently, NADH, sodium‐pumping NADH:ubiquinone oxidoreductase (Na(+)‐NQR) system, membrane potential and intracellular gentamicin were decreased in VA‐RGEN. These findings indicate that the reduced redox state contributes to antibiotic resistance. Interestingly, exogenous glucose potentiated gentamicin to efficiently kill VA‐RGEN through the promotion of the P cycle, NADH, membrane potential and intracellular gentamicin. The potentiation was further confirmed in a zebrafish model. These results indicate that the gentamicin resistance reduces the P cycle and Na(+)‐NQR system and thereby decreases redox state, membrane potential and gentamicin uptake, which can be reversed by exogenous glucose.