Multiomics analysis revealed anti-freezing mechanism of Staphylococcus aureus in its anti-freezing strain

Frozen food is currently a common food type. However, the presence of Staphylococcus aureus contamination caused serious challenge to frozen food safety. In this study, we explored the differences between sensitive strains and anti-freeze strains through multiomics analysis such as proteomics, phosphorylated proteomics, and metabolomics studies to understand the anti-freezing mechanism of S. aureus. This study compared the proteomics, phosphorylated proteomics and metabolomic differences between anti-freeze strains and sensitive strains before and after freezing. pre- and post-freezing, antifreeze strains mainly responded to low temperatures by enhancing antioxidant capacity, energy metabolism and antibiotic synthesis before freezing, while after freezing, bacteria depended more on biofilm formation, nitrogen metabolism and amino acid metabolism to improve frost resistance and adaptability. The ket gene such as sucD and coaD in the energy metabolism and metabolic pathways, gapA1 and tpiA in glucose metabolism and energy balance, asd and gnd in the metabolic pathways of amino acid synthesis and protein synthesis, crtN in the context of protective mechanisms and stress response should be explored furthermore. In metabolomic research, the arginine synthesis pathway, biofilm formation pathway, Adenosine triphosphate-binding cassette transporters and other metabolic pathways deserve special attention and research. IMPORTANCEThis study highlights the importance of understanding the anti-freezing mechanisms of *Staphylococcus aureus* in frozen food safety. By analyzing proteomics, phosphorylated proteomics, and metabolomics, it identifies key pathways like energy metabolism, biofilm formation, and amino acid synthesis that enhance bacterial frost resistance, offering insights for improving food preservation and safety.