Antibiotic tolerance often involves reversible metabolic reprogramming. Here, we applied DIA-based quantitative proteomics to profile Vibrio alginolyticus (V. alginolyticus) under norfloxacin (NOR) exposure and observed widespread decreases in protein abundance, consistent with a global metabolic downshift. The most prominent changes were in central carbon and energy metabolism, encompassing the Embden–Meyerhof–Parnas (EMP), pentose phosphate (PPP), and tricarboxylic acid (TCA) pathways, alongside amino acid metabolism and oxidative phosphorylation. . The study utilizes advancedproteomics techniques, including LC-/MS/MS,to uncoverdifferential protein expression patterns.