Proteomic Analyses of Chlorhexidine Tolerance Mechanisms in <named-content content-type="genus-species">Delftia acidovorans</named-content> Biofilms

ABSTRACT Protein expression and fatty acid profiles of biofilm cells of chlorhexidine-tolerant Delftia acidovorans (MIC = 15 µg/ml) and its chlorhexidine-susceptible mutant (MIC = 1 µg/ml) were investigated. The chlorhexidine-susceptible mutant (MT51) was derived from the parental strain (WT15) usin...

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Autores principales:	Tara Rema, Prabhakara Medihala, John R. Lawrence, Sinisa Vidovic, Gary G. Leppard, Marcia Reid, Darren R. Korber
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Delftia acidovorans tolQ biofilms chlorhexidine tolerance protein expression Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/e6fa31c7a8f9488ebdaacf9d5e4baf09
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spelling	oai:doaj.org-article:e6fa31c7a8f9488ebdaacf9d5e4baf092021-11-15T15:21:37ZProteomic Analyses of Chlorhexidine Tolerance Mechanisms in <named-content content-type="genus-species">Delftia acidovorans</named-content> Biofilms10.1128/mSphere.00017-152379-5042https://doaj.org/article/e6fa31c7a8f9488ebdaacf9d5e4baf092016-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00017-15https://doaj.org/toc/2379-5042ABSTRACT Protein expression and fatty acid profiles of biofilm cells of chlorhexidine-tolerant Delftia acidovorans (MIC = 15 µg/ml) and its chlorhexidine-susceptible mutant (MIC = 1 µg/ml) were investigated. The chlorhexidine-susceptible mutant (MT51) was derived from the parental strain (WT15) using Tn5 transposon mutagenesis. The disrupted gene was identified as tolQ, a component of the tolQRAB gene cluster known to be involved in outer membrane stability. Proteomic responses of biofilm cells were compared by differential in-gel electrophoresis following exposure to chlorhexidine at sub-MIC (10 µg/ml) and above-MIC (30 µg/ml) concentrations. Numerous changes in protein abundance were observed in biofilm cells following chlorhexidine exposure, suggesting that molecular changes occurred during adaptation to chlorhexidine. Forty proteins showing significant differences (≥1.5-fold; P < 0.05) were identified by mass spectrometry and were associated with various functions, including amino acid and lipid biosynthesis, protein translation, energy metabolism, and stress-related functions (e.g., GroEL, aspartyl/glutamyl-tRNA amidotransferase, elongation factor Tu, Clp protease, and hydroxymyristoyl-ACP dehydratase). Several proteins involved in fatty acid synthesis were affected by chlorhexidine, in agreement with fatty acid analysis, wherein chlorhexidine-induced shifts in the fatty acid profile were observed in the chlorhexidine-tolerant cells, primarily the cyclic fatty acids. Transmission electron microscopy revealed more prominent changes in the cell envelope of chlorhexidine-susceptible MT51 cells. This study suggests that multiple mechanisms involving both the cell envelope (and likely TolQ) and panmetabolic regulation play roles in chlorhexidine tolerance in D. acidovorans. IMPORTANCE Delftia acidovorans has been associated with a number of serious infections, including bacteremia, empyema, bacterial endocarditis, and ocular and urinary tract infections. It has also been linked with a variety of surface-associated nosocomial infections. Biofilm-forming antimicrobial-resistant D. acidovorans strains have also been isolated, including ones displaying resistance to the common broad-spectrum agent chlorhexidine. The mechanisms of chlorhexidine resistance in D. acidovorans are not known; hence, a chlorhexidine-susceptible mutant of the tolerant wild-type strain was obtained using transposon mutagenesis, and the proteome and ultrastructural changes of both strains were compared under chlorhexidine challenge.Tara RemaPrabhakara MedihalaJohn R. LawrenceSinisa VidovicGary G. LeppardMarcia ReidDarren R. KorberAmerican Society for MicrobiologyarticleDelftia acidovoranstolQbiofilmschlorhexidine toleranceprotein expressionMicrobiologyQR1-502ENmSphere, Vol 1, Iss 1 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Delftia acidovorans tolQ biofilms chlorhexidine tolerance protein expression Microbiology QR1-502
spellingShingle	Delftia acidovorans tolQ biofilms chlorhexidine tolerance protein expression Microbiology QR1-502 Tara Rema Prabhakara Medihala John R. Lawrence Sinisa Vidovic Gary G. Leppard Marcia Reid Darren R. Korber Proteomic Analyses of Chlorhexidine Tolerance Mechanisms in <named-content content-type="genus-species">Delftia acidovorans</named-content> Biofilms
description	ABSTRACT Protein expression and fatty acid profiles of biofilm cells of chlorhexidine-tolerant Delftia acidovorans (MIC = 15 µg/ml) and its chlorhexidine-susceptible mutant (MIC = 1 µg/ml) were investigated. The chlorhexidine-susceptible mutant (MT51) was derived from the parental strain (WT15) using Tn5 transposon mutagenesis. The disrupted gene was identified as tolQ, a component of the tolQRAB gene cluster known to be involved in outer membrane stability. Proteomic responses of biofilm cells were compared by differential in-gel electrophoresis following exposure to chlorhexidine at sub-MIC (10 µg/ml) and above-MIC (30 µg/ml) concentrations. Numerous changes in protein abundance were observed in biofilm cells following chlorhexidine exposure, suggesting that molecular changes occurred during adaptation to chlorhexidine. Forty proteins showing significant differences (≥1.5-fold; P < 0.05) were identified by mass spectrometry and were associated with various functions, including amino acid and lipid biosynthesis, protein translation, energy metabolism, and stress-related functions (e.g., GroEL, aspartyl/glutamyl-tRNA amidotransferase, elongation factor Tu, Clp protease, and hydroxymyristoyl-ACP dehydratase). Several proteins involved in fatty acid synthesis were affected by chlorhexidine, in agreement with fatty acid analysis, wherein chlorhexidine-induced shifts in the fatty acid profile were observed in the chlorhexidine-tolerant cells, primarily the cyclic fatty acids. Transmission electron microscopy revealed more prominent changes in the cell envelope of chlorhexidine-susceptible MT51 cells. This study suggests that multiple mechanisms involving both the cell envelope (and likely TolQ) and panmetabolic regulation play roles in chlorhexidine tolerance in D. acidovorans. IMPORTANCE Delftia acidovorans has been associated with a number of serious infections, including bacteremia, empyema, bacterial endocarditis, and ocular and urinary tract infections. It has also been linked with a variety of surface-associated nosocomial infections. Biofilm-forming antimicrobial-resistant D. acidovorans strains have also been isolated, including ones displaying resistance to the common broad-spectrum agent chlorhexidine. The mechanisms of chlorhexidine resistance in D. acidovorans are not known; hence, a chlorhexidine-susceptible mutant of the tolerant wild-type strain was obtained using transposon mutagenesis, and the proteome and ultrastructural changes of both strains were compared under chlorhexidine challenge.
format	article
author	Tara Rema Prabhakara Medihala John R. Lawrence Sinisa Vidovic Gary G. Leppard Marcia Reid Darren R. Korber
author_facet	Tara Rema Prabhakara Medihala John R. Lawrence Sinisa Vidovic Gary G. Leppard Marcia Reid Darren R. Korber
author_sort	Tara Rema
title	Proteomic Analyses of Chlorhexidine Tolerance Mechanisms in <named-content content-type="genus-species">Delftia acidovorans</named-content> Biofilms
title_short	Proteomic Analyses of Chlorhexidine Tolerance Mechanisms in <named-content content-type="genus-species">Delftia acidovorans</named-content> Biofilms
title_full	Proteomic Analyses of Chlorhexidine Tolerance Mechanisms in <named-content content-type="genus-species">Delftia acidovorans</named-content> Biofilms
title_fullStr	Proteomic Analyses of Chlorhexidine Tolerance Mechanisms in <named-content content-type="genus-species">Delftia acidovorans</named-content> Biofilms
title_full_unstemmed	Proteomic Analyses of Chlorhexidine Tolerance Mechanisms in <named-content content-type="genus-species">Delftia acidovorans</named-content> Biofilms
title_sort	proteomic analyses of chlorhexidine tolerance mechanisms in <named-content content-type="genus-species">delftia acidovorans</named-content> biofilms
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/e6fa31c7a8f9488ebdaacf9d5e4baf09
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Proteomic Analyses of Chlorhexidine Tolerance Mechanisms in <named-content content-type="genus-species">Delftia acidovorans</named-content> Biofilms

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