This project investigates the role of intestinal β-galactosidase in sepsis using 16S rRNA sequencing, metabolomic, and proteomic analyses. Septic patients showed reduced intestinal β-galactosidase activity associated with depletion of putative β-galactosidase-producing bacteria, including Blautia, Faecalibacterium, and Ruminococcus. Metabolomic profiling identified hesperetin as a key microbiota-associated metabolite, and proteomic analysis identified CDK13 as a critical downstream target regulating NLRP3 inflammasome priming and macrophage pyroptosis. Restoration of intestinal β-galactosidase promoted dietary hesperetin liberation and improved survival in septic mice.