Quantitative Proteomic Analysis on the Slightly Acidic Electrolyzed Water Triggered Viable but Non-Culturable Listeria monocytogenes
Abstract
:1. Introduction
2. Results
2.1. The Culturability of L. monocytogenes after Treatment with Slightly Acidic Electrolyzed Water (SAEW)
2.2. Determination of Cell Viability of L. monocytogenes Using Flow Cytometry
2.3. Morphological Changes of VBNC L. monocytogenes Using Scanning Electron Microscopy
2.4. Differential Expressed Proteins (DEPs) of L. monocytogenes after Treating with SAEW
2.5. GO and KEGG Functional Enrichment Analysis of SAEW-Regulated Proteins
2.6. SAEW-Mediated Protein Chlorination in VBNC L. monocytogenes
3. Discussion
4. Materials and Methods
4.1. Bacterial Cultures and Preparations
4.2. Preparation of Slightly Acidic Electrolyzed Water (SAEW)
4.3. Sample Treatment
4.4. Flow Cytometry Analysis of Treated Samples
4.5. Scanning Electron Microscopy (SEM) Analysis
4.6. Whole-Protein Extraction for Proteomic Analysis
4.7. Mass Spectrometric Analysis
4.7.1. In Solution Digestion
4.7.2. Tandem Mass Tags (TMT) Labeling
4.7.3. SDB Fractionation and Desalted
4.7.4. LC-MS/MS
4.7.5. Data Analysis and Protein Network Construction
4.8. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Chlorine Concentration (mg/L) | Recovery on BHIA (log CFU/mL) | Reduction on BHIA (log CFU/mL) |
---|---|---|
Control | 8.26 ± 0.09 | - |
4 | 7.95 ± 0.17 | 0.31 |
6 | 7.54 ± 0.44 | 0.72 |
8 | ND | 8.26 |
10 | ND | 8.26 |
20 | ND | 8.26 |
Protein IDs | Identified Sequence | Gene Symbol Cl Residue | Fold Change (log2) | |
---|---|---|---|---|
SA20_CTL | SA10_CTL | |||
Q8Y5V1 | EFVPLLVYSPR | deoB, Y350 | 1.038 | 1.102 |
P0DJM2 | IINEPTAAALAYGMDK | dnaK, Y153 | 1.180 | 1.155 |
Q8Y8G1 | MDDLYSEFGEQMDEVAER | dps, Y50 | 0.884 | 0.590 |
P64074 | TSEEMVTWYEEMITK | eno, W273 | 1.809 | 0.564 |
P64074 | TSEEMVTWYEEMITK | eno, Y274 | 1.500 | 1.024 |
Q8Y7G7 | LSVDYYGAATPVNQMASISVPEAR | frr, Y45 | 0.869 | 1.217 |
Q8Y421 | GVYTMQFDHYEEVPK | fusA, Y666 | 2.352 | 2.508 |
Q8Y6D3 | INDLGLPAYDAMVLTLTK | gatB, Y313 | 1.455 | 1.589 |
Q8Y571 | VIPYWMDTIAPEIK | gpmA, W162 | 1.282 | 1.615 |
B8DH59 | HEAVGVGFNAANGEWINMIDAGIVDPTK | groEL, W482 | 1.543 | 1.041 |
B8DH59 | GYTSPYMVTDSDK | groEL, Y201 | 1.044 | 1.056 |
A0A0H3GEZ8 | YGLIETN | hpf, Y181 | 0.882 | 0.951 |
A0A0H3GEZ8 | GENIEVTEPIRDYVEK | hpf, Y20 | 1.310 | 0.927 |
C1KV98 | WDVADLMDYIK | ispH, W175; Y183 | 0.334 | 0.587 |
Q8YAE0 | LAEKYGVSIR | lmo0191, Y143 | −0.053 | 0.076 |
Q8Y8D7 | KYMITSK | nadK1, Y3 | 0.262 | 0.292 |
Q8Y6C8 | VTAEGYVILNK | pcrB, Y112 | 2.267 | 2.457 |
Q71XR3 | NFGAPYELIK | pdxS, Y194 | 1.158 | 1.349 |
P58641 | ANDPVAAYNQVLKEWNA | pyrF, Y224; W231 | −0.504 | −0.326 |
Q927L9 | EQLIFPEIDYDQVSK | rplE, Y143 | −0.272 | −0.532 |
Q8Y6T6 | RPYAPGQHGPTQR | rpsD, Y31 | 1.469 | 0.732 |
Q8Y6T6 | HMYGLTER | rpsD, Y61 | 1.781 | 1.652 |
Q71WF0 | HVPVYVQEDMVGHK | rpsS, Y61 | 0.790 | 1.068 |
Q8Y6U0 | YSGQVQMHIVPFTEIQEVIK | thiI, Y233 | 0.446 | 0.549 |
Q8Y6M7 | TEMPNGQTVQDYITEAITK | tsf, Y125 | 1.771 | 1.791 |
Q8Y422 | QVGVPYIVVFMNK | tuf, Y130 | 3.229 | 2.687 |
Q8Y422 | DLLTEYEFPGDDIPVIK | tuf, Y161 | 1.010 | 0.921 |
Q8Y422 | IDELMEAVDSYIPTPER | tuf, Y199 | 1.353 | 1.216 |
Q8Y422 | LLDYAEAGDNIGALLR | tuf, Y269 | 0.157 | 0.229 |
Q8Y422 | GYADAQAYDQIDGAPEER | tuf, Y41 | 1.229 | 1.230 |
Q8Y422 | HYAHVDCPGHADYVK | tuf, Y88 | 0.538 | 1.220 |
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Chang, H.-Y.; Gui, C.-Y.; Huang, T.-C.; Hung, Y.-C.; Chen, T.-Y. Quantitative Proteomic Analysis on the Slightly Acidic Electrolyzed Water Triggered Viable but Non-Culturable Listeria monocytogenes. Int. J. Mol. Sci. 2023, 24, 10616. https://doi.org/10.3390/ijms241310616
Chang H-Y, Gui C-Y, Huang T-C, Hung Y-C, Chen T-Y. Quantitative Proteomic Analysis on the Slightly Acidic Electrolyzed Water Triggered Viable but Non-Culturable Listeria monocytogenes. International Journal of Molecular Sciences. 2023; 24(13):10616. https://doi.org/10.3390/ijms241310616
Chicago/Turabian StyleChang, Hsin-Yi, Chin-Ying Gui, Tsui-Chin Huang, Yen-Con Hung, and Tai-Yuan Chen. 2023. "Quantitative Proteomic Analysis on the Slightly Acidic Electrolyzed Water Triggered Viable but Non-Culturable Listeria monocytogenes" International Journal of Molecular Sciences 24, no. 13: 10616. https://doi.org/10.3390/ijms241310616