TMT proteomic analysis for molecular mechanism of Staphylococcus aureus in response to freezing stress

The foodborne pathogen Staphylococcus aureus continues to challenge the food industry due to the pathogenicity and tolerance of the bacterium. As a common storage condition for frozen food during transportation, distribution, and storage, freezing does not seem to be entirely safe due to the cold tolerance of S. aureus. In addition, our study indicated that the biofilm formation ability of S. aureus was significantly increased in response to freezing stress. To explore the molecular mechanism regulating the response to freezing stress, the proteomics signature of S. aureus after freezing stress based on tandem mass tag (TMT) labeling and liquid chromatography tandem mass spectrometry (LC-MS/MS) was analyzed. Gene Ontology and pathway analysis revealed that ribosome function, metabolism, RNA repair, and stress response proteins were differentially regulated (P < 0.05). Furthermore, transpeptidase sortase A, biofilm operon icaADBC HTH-type negative transcriptional regulator IcaR, and HTH-type transcriptional regulator MgrA were involved in the modulation of increased biofilm formation in response to freezing stress (P < 0.05). Moreover, significant lysine acetylation and malonylation signals in the S. aureus response to freezing stress were observed. Collectively, the current work provides additional insight for comprehending the molecular mechanism of S. aureus in response to freezing stress and presents potential targets for developing strategies to control S. aureus. KEY POINTS: • TMT proteomic analysis was first used on S. aureus in response to freezing stress. • Ribosome-, metabolism-, and biofilm-related proteins change after freezing stress. • Increased biofilm formation in S. aureus responded to freezing stress.