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Naphthalene uptake by a Pseudomonas fluorescens isolate.

B E Whitman1, D R Lueking, J R Mihelcic

  • 1Department of GeoEnvironmental Science and Engineering, Wilkes University, Wilkes-Barre, PA 18766, USA.

Canadian Journal of Microbiology
|February 25, 1999
PubMed
Summary

Pseudomonas fluorescens actively transports naphthalene into its cells via a specific, energy-dependent system. This uptake is crucial for naphthalene utilization and is inhibited by certain chemicals and naphthalene analogues.

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Area of Science:

  • Microbiology
  • Biochemistry
  • Environmental Science

Background:

  • Naphthalene is an aromatic hydrocarbon that can be utilized by various microorganisms.
  • Understanding the initial steps of xenobiotic compound uptake is crucial for bioremediation and microbial metabolism studies.

Purpose of the Study:

  • To investigate the mechanism of naphthalene uptake in Pseudomonas fluorescens.
  • To characterize the kinetics and specificity of the naphthalene transport system.

Main Methods:

  • Utilized [1-14C]naphthalene to quantify uptake in Pseudomonas fluorescens.
  • Tested inhibition by metabolic inhibitors (azide, 2,4-dinitrophenol) and structural analogues (alpha-naphthol).
  • Assessed naphthalene uptake in a naphthalene-nonutilizing mutant strain (TG-5).

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Main Results:

  • Naphthalene uptake exhibited Michaelis-Menten kinetics with an affinity constant (Kt) of 11 microM and maximal velocity (Vmax) of 17 nmol.h-1.mg-1.
  • Uptake was significantly inhibited by metabolic inhibitors and structural analogues, and absent in the mutant strain.
  • Alpha-naphthol noncompetitively inhibited uptake with a Ki of 0.041 microM; intracellular ATP levels remained unaffected by inhibitors.

Conclusions:

  • Pseudomonas fluorescens possesses a specific, energy-dependent transport system for naphthalene.
  • This transport system represents the initial and critical step in naphthalene utilization by the bacterium.