title: Growth media simulating ileal and colonic environments affect the intracellular proteome and carbon fluxes of enterohemorrhagic Escherichia coli O157:H7 strain EDL933.
abstract: In this study, the intracellular proteome of Escherichia coli O157:H7 strain EDL933 was analyzed by two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) spectrometry after growth in simulated ileal environment media (SIEM) and simulated colonic environment media (SCEM) under aerobic and microaerobic conditions. Differentially expressed intracellular proteins were identified and allocated to functional protein groups. Moreover, metabolic fluxes were analyzed by isotopologue profiling with [U-(13)C(6)]glucose as a tracer. The results of this study show that EDL933 responds with differential expression of a complex network of proteins and metabolic pathways, reflecting the high metabolic adaptability of the strain. Growth in SIEM and SCEM is obviously facilitated by the upregulation of nucleotide biosynthesis pathway proteins and could be impaired by exposition to 50 µM 6-mercaptopurine under aerobic conditions. Notably, various stress and virulence factors, including Shiga toxin, were expressed without having contact with a human host.
authors.affiliation: Institute of Food Science and Biotechnology, Department of Food Microbiology, University of Hohenheim, Stuttgart, Germany.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Polzin
authors.forename: Claudia
authors.initials: C
authors.lastname: Huber
authors.forename: Eva
authors.initials: E
authors.lastname: Eylert
authors.forename: Ines
authors.initials: I
authors.lastname: Elsenhans
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Eisenreich
authors.forename: Herbert
authors.initials: H
authors.lastname: Schmidt
mesh_terms: D002247:Carbon Isotopes / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D019453:Escherichia coli O157 / Q000187:drug effects* / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D007422:Intestines / Q000737:chemistry; D053858:Metabolic Networks and Pathways / Q000187:drug effects* / Q000502:physiology; D010449:Peptide Mapping; D020543:Proteome / Q000187:drug effects*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D002247:Carbon Isotopes; D003470:Culture Media; D020543:Proteome
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