Inactivation of the Pseudomonas putida KT2440 dsbA gene promotes extracellular matrix production and biofilm formation

Yunho Lee, Sejong Oh, Woojun Park

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

To identify genes essential to biofilm formation in Pseudomonas putida KT2440, 12 mutants defective in oxidative stress-related or metabolic pathway-related genes were evaluated. Of them, only the dsbA mutant lacking the disulfide bond isomerase exhibited significantly increased attachment to the polystyrene surface. Visual evaluation by extracellular matrix staining and scanning electron microscopy indicated that the KT2440-ΔdsbA strain displays enhanced extracellular matrix production, rugose colony morphology on agar plates and floating pellicles in static culture. Accordingly, we propose that deletion of the dsbA gene may stimulate production of the extracellular matrix, resulting in those phenotypes. In addition, the lack of detectable fluorescence in the KT2440-ΔdsbA under UV light as well as in both the wild type and the KT2440-ΔdsbA when grown on Luria-Bertani plates containing ferrous iron suggests that the fluorescent molecule may be a fluorescent siderophore with its synthesis/secretion controlled by DsbA in KT2440. These phenotypic defects observed in the dsbA mutant were complemented by the full-length KT2440 and Escherichia coli dsbA genes. In contrast to the role of DsbA in other bacteria, our results provide the first evidence that disruption of P. putida KT2440 dsbA gene overproduces the extracellular matrix and thus promotes biofilm formation.

Original languageEnglish
Pages (from-to)38-48
Number of pages11
JournalFEMS Microbiology Letters
Volume297
Issue number1
DOIs
Publication statusPublished - 2009 Aug 1

Fingerprint

Pseudomonas putida
Biofilms
Extracellular Matrix
Genes
Protein Disulfide-Isomerases
Siderophores
Polystyrenes
Essential Genes
Gene Deletion
Ultraviolet Rays
Metabolic Networks and Pathways
Electron Scanning Microscopy
Agar
Oxidative Stress
Iron
Fluorescence
Staining and Labeling
Escherichia coli
Bacteria
Phenotype

Keywords

  • Escherichia coli O157
  • Motility assay
  • Oxidative stress
  • Pellicle
  • Pleiotropic phenotypes
  • Soil bacterium

ASJC Scopus subject areas

  • Microbiology
  • Genetics
  • Molecular Biology

Cite this

Inactivation of the Pseudomonas putida KT2440 dsbA gene promotes extracellular matrix production and biofilm formation. / Lee, Yunho; Oh, Sejong; Park, Woojun.

In: FEMS Microbiology Letters, Vol. 297, No. 1, 01.08.2009, p. 38-48.

Research output: Contribution to journalArticle

@article{aa99153192fe4d1a8abd42ae45e26b22,
title = "Inactivation of the Pseudomonas putida KT2440 dsbA gene promotes extracellular matrix production and biofilm formation",
abstract = "To identify genes essential to biofilm formation in Pseudomonas putida KT2440, 12 mutants defective in oxidative stress-related or metabolic pathway-related genes were evaluated. Of them, only the dsbA mutant lacking the disulfide bond isomerase exhibited significantly increased attachment to the polystyrene surface. Visual evaluation by extracellular matrix staining and scanning electron microscopy indicated that the KT2440-ΔdsbA strain displays enhanced extracellular matrix production, rugose colony morphology on agar plates and floating pellicles in static culture. Accordingly, we propose that deletion of the dsbA gene may stimulate production of the extracellular matrix, resulting in those phenotypes. In addition, the lack of detectable fluorescence in the KT2440-ΔdsbA under UV light as well as in both the wild type and the KT2440-ΔdsbA when grown on Luria-Bertani plates containing ferrous iron suggests that the fluorescent molecule may be a fluorescent siderophore with its synthesis/secretion controlled by DsbA in KT2440. These phenotypic defects observed in the dsbA mutant were complemented by the full-length KT2440 and Escherichia coli dsbA genes. In contrast to the role of DsbA in other bacteria, our results provide the first evidence that disruption of P. putida KT2440 dsbA gene overproduces the extracellular matrix and thus promotes biofilm formation.",
keywords = "Escherichia coli O157, Motility assay, Oxidative stress, Pellicle, Pleiotropic phenotypes, Soil bacterium",
author = "Yunho Lee and Sejong Oh and Woojun Park",
year = "2009",
month = "8",
day = "1",
doi = "10.1111/j.1574-6968.2009.01650.x",
language = "English",
volume = "297",
pages = "38--48",
journal = "FEMS Microbiology Letters",
issn = "0378-1097",
publisher = "Wiley-Blackwell",
number = "1",

}

TY - JOUR

T1 - Inactivation of the Pseudomonas putida KT2440 dsbA gene promotes extracellular matrix production and biofilm formation

AU - Lee, Yunho

AU - Oh, Sejong

AU - Park, Woojun

PY - 2009/8/1

Y1 - 2009/8/1

N2 - To identify genes essential to biofilm formation in Pseudomonas putida KT2440, 12 mutants defective in oxidative stress-related or metabolic pathway-related genes were evaluated. Of them, only the dsbA mutant lacking the disulfide bond isomerase exhibited significantly increased attachment to the polystyrene surface. Visual evaluation by extracellular matrix staining and scanning electron microscopy indicated that the KT2440-ΔdsbA strain displays enhanced extracellular matrix production, rugose colony morphology on agar plates and floating pellicles in static culture. Accordingly, we propose that deletion of the dsbA gene may stimulate production of the extracellular matrix, resulting in those phenotypes. In addition, the lack of detectable fluorescence in the KT2440-ΔdsbA under UV light as well as in both the wild type and the KT2440-ΔdsbA when grown on Luria-Bertani plates containing ferrous iron suggests that the fluorescent molecule may be a fluorescent siderophore with its synthesis/secretion controlled by DsbA in KT2440. These phenotypic defects observed in the dsbA mutant were complemented by the full-length KT2440 and Escherichia coli dsbA genes. In contrast to the role of DsbA in other bacteria, our results provide the first evidence that disruption of P. putida KT2440 dsbA gene overproduces the extracellular matrix and thus promotes biofilm formation.

AB - To identify genes essential to biofilm formation in Pseudomonas putida KT2440, 12 mutants defective in oxidative stress-related or metabolic pathway-related genes were evaluated. Of them, only the dsbA mutant lacking the disulfide bond isomerase exhibited significantly increased attachment to the polystyrene surface. Visual evaluation by extracellular matrix staining and scanning electron microscopy indicated that the KT2440-ΔdsbA strain displays enhanced extracellular matrix production, rugose colony morphology on agar plates and floating pellicles in static culture. Accordingly, we propose that deletion of the dsbA gene may stimulate production of the extracellular matrix, resulting in those phenotypes. In addition, the lack of detectable fluorescence in the KT2440-ΔdsbA under UV light as well as in both the wild type and the KT2440-ΔdsbA when grown on Luria-Bertani plates containing ferrous iron suggests that the fluorescent molecule may be a fluorescent siderophore with its synthesis/secretion controlled by DsbA in KT2440. These phenotypic defects observed in the dsbA mutant were complemented by the full-length KT2440 and Escherichia coli dsbA genes. In contrast to the role of DsbA in other bacteria, our results provide the first evidence that disruption of P. putida KT2440 dsbA gene overproduces the extracellular matrix and thus promotes biofilm formation.

KW - Escherichia coli O157

KW - Motility assay

KW - Oxidative stress

KW - Pellicle

KW - Pleiotropic phenotypes

KW - Soil bacterium

UR - http://www.scopus.com/inward/record.url?scp=67650723517&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67650723517&partnerID=8YFLogxK

U2 - 10.1111/j.1574-6968.2009.01650.x

DO - 10.1111/j.1574-6968.2009.01650.x

M3 - Article

C2 - 19500143

AN - SCOPUS:67650723517

VL - 297

SP - 38

EP - 48

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

SN - 0378-1097

IS - 1

ER -