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Effects of outer membrane vesicle formation, surface-layer production and nanopod development on the metabolism of phenanthrene by Delftia acidovorans Cs1-4.

ABSTRACT: Nanopods are extracellular structures arising from the convergence of two widely distributed bacterial characteristics: production of outer membrane vesicles (OMV) and formation of surface layers (S-layers). Nanopod production is driven by OMV formation, and in Delftia acidovorans Cs1-4 growth on phenanthrene induces OMV/nanopod formation. While OMV production has been associated with many functions, particularly with pathogens, linkage to biodegradation has been limited to a membrane stress response to lipophilic compounds. The objectives of this study were to determine: 1.) Whether induction of nanopod formation was linked to phenanthrene metabolism or a non-specific membrane stress response, and 2.) The relative importance of OMV/nanopod formation vs. formation of the S-layer alone to phenanthrene utilization. Membrane stress response was investigated by quantifying nanopod formation following exposure to compounds that exceeded phenanthrene in membrane stress-inducing potential. Naphthalene did not induce nanopod formation, and toluene was a weak inducer compared to phenanthrene (two- vs. six-fold increase, respectively). Induction of nanopod formation by growth on phenanthrene was therefore linked to phenanthrene metabolism and not a membrane stress response. Impacts on phenanthrene biodegradation of OMV/nanopod production vs. S-layer formation were assessed with D. acidovorans Cs1-4 mutants deficient in S-layer formation or OMV/nanopod production. Both mutants had impaired growth on phenanthrene, but the loss of OMV/nanopod production was more significant than loss of the S-layer. The S-layer of D. acidovorans Cs1-4 did not affect phenanthrene uptake, and its primary role in phenanthrene biodegradation process appeared to be enabling nanopod development. Nanopods appeared to benefit phenanthrene biodegradation by enhancing cellular retention of metabolites. Collectively, these studies established that nanopod/OMV formation was an essential characteristic of the D. acidovorans Cs1-4 phenanthrene degradation process. This report thus established a new dimension in the area of biodegradation, namely, the involvement of extracellular structures as elements supporting metabolic processes underlying biodegradation.

SUBMITTER: Shetty A

PROVIDER: S-EPMC3958437 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Effects of outer membrane vesicle formation, surface-layer production and nanopod development on the metabolism of phenanthrene by Delftia acidovorans Cs1-4.

Shetty Ameesha A Hickey William J WJ

PloS one 20140318 3

Nanopods are extracellular structures arising from the convergence of two widely distributed bacterial characteristics: production of outer membrane vesicles (OMV) and formation of surface layers (S-layers). Nanopod production is driven by OMV formation, and in Delftia acidovorans Cs1-4 growth on phenanthrene induces OMV/nanopod formation. While OMV production has been associated with many functions, particularly with pathogens, linkage to biodegradation has been limited to a membrane stress res ...[more]

PMID: 24642639

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Project description: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.

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Dataset Information

Effects of outer membrane vesicle formation, surface-layer production and nanopod development on the metabolism of phenanthrene by Delftia acidovorans Cs1-4.

Publications

Effects of outer membrane vesicle formation, surface-layer production and nanopod development on the metabolism of phenanthrene by Delftia acidovorans Cs1-4.

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