Proteomics-based mechanistic investigation of Staphylococcus aureus inactivation by pulsed electric field

ABSTRACT: The Staphylococcus aureus strain was cultured in LB medium at 150 rpm and 37 ℃ to a final optical density (OD) of 1.0 at 600 n m. Subsequently, 1 m L of the culture broth was collected and centrifuged at 8000 rpm, 4 ℃ for 2 min. The cell pellet was re-suspended in 2 m L of ice-cold 0.1% (w/v) bacteriological peptone solution and divided into 70 µL aliquots. For PEF treatment, each aliquot was pipetted into a sterile 1.0 mm electroporation cuvette and pulsed using a BTX ECM830 Square Wave Electroporation System (BTX, United States). For a cell killing extent of 94.89±0.90% , pulse intensity,number and duration were set to 11 kV·cm-1 ,86 and 70μs，respectively. The killing extent was 50.29±5.27% with the treatment parameters of 9kV·cm-1 , pulse number of 50, pulse duration of 50μs. For controls, a mock treatment without the application of PEF was performed, with identical downstream procedures. The whole experiment described above was conducted in triplicate. Extract the total protein of the sample and perform quality inspection, reductive alkylation and enzymatic hydrolysis, TMT labeling and mixing, and perform two-dimensional analysis by RPLC one-dimensional separation and liquid phase tandem mass spectrometry (Easy-nLC 1200 combined with Q Exactive mass spectrometer). The raw files off the computer are analyzed by ProteomeDiscovererTM Software 2.2, and the searched species is the self-tested Staphylococcus aureus CGMCCC26003 genome database.

ORGANISM(S): Staphylococcus Aureus

SUBMITTER: Yanbo Song  

PROVIDER: PXD026761 | iProX | Thu Jun 17 00:00:00 BST 2021

REPOSITORIES: iProX