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Related Experiment Videos

Nitrosating activity in Escherichia coli.

C M O'Donnell1, C Edwards

  • 1Department of Genetics and Microbiology, The University, Liverpool, UK.

FEMS Microbiology Letters
|August 1, 1992
PubMed
Summary
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Nitrosation activity in Escherichia coli was linked to intact cells and aerobic conditions. NADH-dependent nitrite reductase (nir) appears crucial for this process, as indicated by mutant analysis.

Area of Science:

  • Microbiology
  • Biochemistry
  • Enzymology

Background:

  • Nitrosation is a chemical process with implications in various biological systems.
  • Escherichia coli (E. coli) is a model organism for studying bacterial metabolism and enzyme function.
  • Nitrite reductase (nir) enzymes play a key role in nitrogen metabolism.

Purpose of the Study:

  • To investigate the factors influencing nitrosation activity in E. coli isolates.
  • To determine the role of nitrite reductase (nir) in the nitrosation process.

Main Methods:

  • Measurement of nitrosation activity in intact E. coli cells and various nir mutants.
  • Assessment of activity under different growth conditions (aerobic vs. oxygen-limited).
  • Evaluation of the effect of cell treatments (sonication, osmotic shock) and media components (nitrite) on activity.

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Main Results:

  • Nitrosation activity was detected only in intact E. coli cells and was sensitive to physical disruption.
  • Aerobic growth conditions correlated with higher nitrosation activity compared to oxygen-limited conditions.
  • Two specific nir mutants exhibited a complete loss of nitrosation activity, implicating NADH-dependent nitrite reductase.

Conclusions:

  • NADH-dependent nitrite reductase is directly or indirectly involved in the nitrosation activity observed in E. coli.
  • Cell integrity and aerobic metabolism are important factors for bacterial nitrosation.