Resumen: This work explores the bactericidal effect of (+)-limonene, the major constituent of citrus fruits' essential oils, against E. coli. The degree of E. coli BJ4 inactivation achieved by (+)-limonene was influenced by the pH of the treatment medium, being more bactericidal at pH 4.0 than at pH 7.0. Deletion of rpoS and exposure to a sub-lethal heat or an acid shock did not modify E. coli BJ4 resistance to (+)-limonene. However, exposure to a sub-lethal cold shock decreased its resistance to (+)-limonene. Although no sub-lethal injury was detected in the cell envelopes after exposure to (+)-limonene by the selective-plating technique, the uptake of propidium iodide by inactivated E. coli BJ4 cells pointed out these structures as important targets in the mechanism of action. Attenuated Total Reflectance Infrared Microspectroscopy (ATR-IRMS) allowed identification of altered E. coli BJ4 structures after (+)-limonene treatments as a function of the treatment pH: ß-sheet proteins at pH 4.0 and phosphodiester bonds at pH 7.0. The increased sensitivity to (+)-limonene observed at pH 4.0 in an E. coli MC4100 lptD4213 mutant with an increased outer membrane permeability along with the identification of altered ß-sheet proteins by ATR-IRMS indicated the importance of this structure in the mechanism of action of (+)-limonene. The study of mechanism of inactivation by (+)-limonene led to the design of a synergistic combined process with heat for the inactivation of the pathogen E. coli O157:H7 in fruit juices. These results show the potential of (+)-limonene in food preservation, either acting alone or in combination with lethal heat treatments. Idioma: Inglés DOI: 10.1371/journal.pone.0056769 Año: 2013 Publicado en: PLoS ONE 8, 2 (2013), e56769 [10 pp] ISSN: 1932-6203 Factor impacto JCR: 3.534 (2013) Categ. JCR: MULTIDISCIPLINARY SCIENCES rank: 8 / 56 = 0.143 (2013) - Q1 - T1 Financiación: info:eu-repo/grantAgreement/ES/MINECO/AGL2009-11660 Financiación: info:eu-repo/grantAgreement/ES/MINECO/AGL2012-32165 Tipo y forma: Artículo (Versión definitiva) Área (Departamento): Área Tecnología de Alimentos (Dpto. Produc.Animal Cienc.Ali.)