title: Carvacrol induces heat shock protein 60 and inhibits synthesis of flagellin in Escherichia coli O157:H7.
abstract: The essential oils of oregano and thyme are active against a number of food-borne pathogens, such as Escherichia coli O157:H7. Carvacrol is one of the major antibacterial components of these oils, and p-cymene is thought to be its precursor in the plant. The effects of carvacrol and p-cymene on protein synthesis in E. coli O157:H7 ATCC 43895 cells were investigated. Bacteria were grown overnight in Mueller-Hinton broth with a sublethal concentration of carvacrol or p-cymene, and their protein compositions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and confirmed by Western blotting. The presence of 1 mM carvacrol during overnight incubation caused E. coli O157:H7 to produce significant amounts of heat shock protein 60 (HSP60) (GroEL) (P < 0.05) and inhibited the synthesis of flagellin highly significantly (P < 0.001), causing cells to be aflagellate and therefore nonmotile. The amounts of HSP70 (DnaK) were not significantly affected. p-Cymene at 1 mM or 10 mM did not induce HSP60 or HSP70 in significant amounts and did not have a significant effect on flagellar synthesis. Neither carvacrol (0.3, 0.5, 0.8, or 1 mM) nor p-cymene (0.3, 0.5, or 0.8 mM) treatment of cells in the mid-exponential growth phase induced significant amounts of HSP60 or HSP70 within 3 h, although numerical increases of HSP60 were observed. Motility decreased with increasing concentrations of both compounds, but existing flagella were not shed. This study is the first to demonstrate that essential oil components induce HSP60 in bacteria and that overnight incubation with carvacrol prevents the development of flagella in E. coli O157:H7.
authors.affiliation: Institute for Risk Assessment Sciences, Division of Veterinary Public Health, Faculty of Veterinary Medicine, Utrecht University, 3508TD Utrecht, The Netherlands. s.a.burt@iras.uu.nl
authors.forename: Sara A
authors.initials: SA
authors.lastname: Burt
authors.forename: Ruurd
authors.initials: R
authors.lastname: van der Zee
authors.forename: Ad P
authors.initials: AP
authors.lastname: Koets
authors.forename: Anko M
authors.initials: AM
authors.lastname: de Graaff
authors.forename: Frans
authors.initials: F
authors.lastname: van Knapen
authors.forename: Wim
authors.initials: W
authors.lastname: Gaastra
authors.forename: Henk P
authors.initials: HP
authors.lastname: Haagsman
authors.forename: Edwin J A
authors.initials: EJ
authors.lastname: Veldhuizen
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D015153:Blotting, Western; D018834:Chaperonin 60 / Q000096:biosynthesis*; D000081007:Cymenes; D004591:Electrophoresis, Polyacrylamide Gel; D019453:Escherichia coli O157 / Q000737:chemistry / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D029968:Escherichia coli Proteins / Q000096:biosynthesis*; D005407:Flagella / Q000737:chemistry; D005408:Flagellin / Q000096:biosynthesis*; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis; D008124:Locomotion; D008857:Microscopy, Interference; D039821:Monoterpenes / Q000494:pharmacology*; D020543:Proteome / Q000032:analysis
chemical_list: D000900:Anti-Bacterial Agents; D018834:Chaperonin 60; D000081007:Cymenes; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D039821:Monoterpenes; D020543:Proteome; D005408:Flagellin; C007210:4-cymene; C073316:carvacrol