Project description:The spores of A. acidoterrestris cryopreserved at -80 ℃ were activated by inoculating in AAM medium and cultured at 45 ℃ to logarithmic phase [19]. The activated bacterial suspension was centrifuged and resuspended into pH 2.0, 2.5, 3.0, and 4.0 (Control) medium, which were correspondingly cultured in medium for 20 and 60 minutes, respectively. Scanning electron microscopy was used to monitor the bacterial activity and morphology different acid stress.Alicyclobacillus acidoterrestris (A. acidoterrestris) causes pasteurized acidic juice spoilage, resulting in a significant decline of juice quality, and causing economic losses. Exploration of A. acidoterrestris in response to acid stress could help control contamination caused by the bacteria. In this study, the mechanism of A. acidoterrestris in response to acid stress was studied by quantitative phosphoproteomics technique. Result showed that the phosphorylation of 40 proteins in A. acidoterrestris were closely related to the regulation of acid stress. The KEGG pathway enrichment analysis showed that the quorum sensing pathway which might involved in the perception of A. acidoterrestris was mainly enriched. we found that the up-regulation of Spo0A and YidC phosphorylation, which may resist acid stress by forming spores. The phosphorylation level of pyruvate kinase increased, which may improve bacterial acid stress resistance through formation of energy supply. The phosphorylation level of ABC transporter permease was significantly up-regulated, which may be part of the cell adaptation adjustment and contribute to the survival of A. acidoterrestris under acid stress. In summary, the molecular mechanism of acid stress regulation of A. acidoterrestris was proposed via quantitative phosphoproteomics, which provided a theoretical and experimental basis for further investigation acid resistance mechanism of A. acidoterrestris.
Project description:Alicyclobacillus acidoterrestris is a major putrefying bacterium that can cause pecuniary losses in the global juice industry. Current detection approaches are time-consuming and exhibit reduced specificity and sensitivity. In this study, an immunoproteomic approach was utilized to identify specific biomarkers from A. acidoterrestris for the development of new detection methods. Cell surface-associated proteins were extracted and separated by 2-D (two-dimensional) gel electrophoresis. Immunogenic proteins were detected by Western blot analysis using antisera against A. acidoterrestris. Twenty-two protein spots exhibiting immunogenicity were excised and eighteen of the associated spots were successfully identified by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF MS). These proteins were observed to be involved in energy and carbohydrate metabolism, transmembrane transport, response to oxidative stress, polypeptide biosynthesis, and molecule binding activity. This is the first report detailing the identification of cell surface-associated antigens of A. acidoterrestris. The identified immunogenic proteins could serve as potential targets for the development of novel detection methods.