title: 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.
authors.affiliation: Key Laboratory of Fruit and Vegetable Processing, Ministry of Agriculture, PR China.|Research Center for Fruit and Vegetable Processing Engineering, Ministry of Education, PR China.|College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, PR China.|State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, Jiangsu, PR China.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Liao
authors.affiliation: Research Center for Fruit and Vegetable Processing Engineering, Ministry of Education, PR China.|Key Laboratory of Fruit and Vegetable Processing, Ministry of Agriculture, PR China.|College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, PR China.
authors.forename: Fusheng
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Research Center for Fruit and Vegetable Processing Engineering, Ministry of Education, PR China.|Key Laboratory of Fruit and Vegetable Processing, Ministry of Agriculture, PR China.|College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, PR China.
authors.forename: Xiaosong
authors.initials: X
authors.lastname: Hu
authors.affiliation: Research Center for Fruit and Vegetable Processing Engineering, Ministry of Education, PR China.|Key Laboratory of Fruit and Vegetable Processing, Ministry of Agriculture, PR China.|College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, PR China.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Liao
mesh_terms: D002245:Carbon Dioxide / Q000494:pharmacology*; D002853:Chromatography, Liquid / Q000379:methods; D004269:DNA, Bacterial / Q000187:drug effects* / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000187:drug effects* / Q000378:metabolism; D006874:Hydrostatic Pressure; D011419:Propidium; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D002245:Carbon Dioxide; D011419:Propidium