000057860 001__ 57860
000057860 005__ 20170327111912.0
000057860 0247_ $$2doi$$a10.1371/journal.pone.0094072
000057860 0248_ $$2sideral$$a91078
000057860 037__ $$aART-2014-91078
000057860 041__ $$aeng
000057860 100__ $$aChueca, Beatriz
000057860 245__ $$aDifferential Mechanism of Escherichia coli Inactivation by (+)-Limonene as a Function of Cell Physiological State and Drug's Concentration
000057860 260__ $$c2014
000057860 5060_ $$aAccess copy available to the general public$$fUnrestricted
000057860 5203_ $$a(+)-limonene is a lipophilic antimicrobial compound, extracted from citrus fruits' essential oils, that is used as a flavouring agent and organic solvent by the food industry. A recent study has proposed a common and controversial mechanism of cell death for bactericidal antibiotics, in which hydroxyl radicals ultimately inactivated cells. Our objective was to determine whether the mechanism of Escherichia coli MG1655 inactivation by (+)-limonene follows that of bactericidal antibiotics. A treatment with 2,000 µL/L (+)-limonene inactivated 4 log10 cycles of exponentially growing E. coli cells in 3 hours. On one hand, an increase of cell survival in the ¿acnB mutant (deficient in a TCA cycle enzyme), or in the presence of 2,2'-dipyridyl (inhibitor of Fenton reaction by iron chelation), thiourea, or cysteamine (hydroxyl radical scavengers) was observed. Moreover, the ¿recA mutant (deficient in an enzyme involved in SOS response to DNA damage) was more sensitive to (+)-limonene. Thus, this indirect evidence indicates that the mechanism of exponentially growing E. coli cells inactivation by 2,000 µL/L (+)-limonene is due to the TCA cycle and Fenton-mediated hydroxyl radical formation that caused oxidative DNA damage, as observed for bactericidal drugs. However, several differences have been observed between the proposed mechanism for bactericidal drugs and for (+)-limonene. In this regard, our results demonstrated that E. coli inactivation was influenced by its physiological state and the drug's concentration: experiments with stationary-phase cells or 4,000 µL/L (+)-limonene uncovered a different mechanism of cell death, likely unrelated to hydroxyl radicals. Our research has also shown that drug's concentration is an important factor influencing the mechanism of bacterial inactivation by antibiotics, such as kanamycin. These results might help in improving and spreading the use of (+)-limonene as an antimicrobial compound, and in clarifying the controversy about the mechanism of inactivation by bactericidal antibiotics.
000057860 536__ $$9info:eu-repo/grantAgreement/ES/MINECO/AGL2012-32165
000057860 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000057860 590__ $$a3.234$$b2014
000057860 591__ $$aMULTIDISCIPLINARY SCIENCES$$b8 / 56 = 0.143$$c2014$$dQ1$$eT1
000057860 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000057860 700__ $$0(orcid)0000-0002-0238-6328$$aPagan, Rafael$$uUniversidad de Zaragoza
000057860 700__ $$aGarcia-Gonzalo, Diego
000057860 7102_ $$12008$$2780$$aUniversidad de Zaragoza$$bDepartamento de Producción Animal y Ciencia de los Alimentos$$cTecnología de Alimentos
000057860 773__ $$g9, 4 (2014), e94072 [7 pp]$$pPLoS One$$tPLoS One$$x1932-6203
000057860 8564_ $$s364003$$uhttp://zaguan.unizar.es/record/57860/files/texto_completo.pdf$$yVersión publicada
000057860 8564_ $$s126339$$uhttp://zaguan.unizar.es/record/57860/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000057860 909CO $$ooai:zaguan.unizar.es:57860$$particulos$$pdriver
000057860 951__ $$a2016-12-14-14:51:44
000057860 980__ $$aARTICLE