Abuse　of　antibiotics　makes　antibiotic-resistance　become　a　huge　challenge　in　bacterial　infection　treatment.　The　discovery　of　new　antibiotics　is　of　great　significance　to　human　health.　In　this　study,　the　antibacterial　mechanism　of　Sparassis　crispa　polysaccharides　(SCPs)　was　explored.　The　SCPs　isolated　from　Sparassis　crispa　was　composed　of　fucose,　glucose　and　galactose　with　a　molar　ratio　of　0.043　:　0.652:　0.305.　Bacteriostatic　tests　showed　SCPs　inhibited　the　growth　of　Staphylococcus　aureus　better　than　Escherichia　coli＇s,　and　damage　to　bacteria　was　observed　under　scanning　electron　microscopy.　Metabolomic　analysis　based　on　HPLC-Q-TOF/MS　indicated　that　SCPs　disrupted　metabolism　of　the　glycolysis　and　tricarboxylic　acid　cycle　pathways　in　S.　aureus.　The　variations　of　fructose-1,6-diphosphate,　1,3-diphosphoglycerol,　succinate　and　oxaloacetate　were　significant,　whose　systematic　changes　accompanied　with　decrease　of　ATP　in　cells　indicated　that　SCPs　could　exert　antibacterial　effects　by　inducing　dysfunction　of　catabolism　and　energy　metabolism.　Our　research　confirmed　the　antibacterial　properties　of　SCPs　and　provided　a　perspective　for　understanding　antibacterial　mechanism　of　polysaccharides　from　natural　products　through　metabolomics　technology.　©　2021