Effects of high-pressure carbon dioxide on proteins and DNA in Escherichia coli

Abstract

Protein changes in Escherichia coli, when subjected to high-pressure carbon dioxide (HPCD) at 10 MPa and 3 °C for 5–75 min, were assessed using the Bradford method, 2D electrophoresis (2-DE) and liquid chromatography-electrospray ionization-MS-MS (LC-ESI-MS-MS). The changes in DNA in E. coli under the same conditions were also investigated by using flow cytometry with propidium iodide and acridine orange, agarose gel electrophoresis (AGE) and the comet assay. The results showed that HPCD induced leakage loss of the proteins and DNA of E. coli as a function of treatment time. With regard to the protein changes, 182 proteins in the 2-DE profile were not found in the HPCD-treated E. coli. Among 20 selected protein spots exhibiting significant changes in intensity, 18 protein spots were identified as 15 known proteins and two as hypothetical proteins. These proteins were involved in cell composition, energy metabolism pathways, nucleic acid metabolism, global stress regulation and general metabolism. The DNA denaturation of E. coli induced by HPCD was demonstrated in this study for the first time to our knowledge, and the denaturation was enhanced by increasing treatment time. However, HPCD did not cause DNA degradation, as suggested by both AGE analysis and the comet assay.