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Keywords:

  • nitrate reduction;
  • nitrite conversion;
  • nitric oxide;
  • nitrosative stress

Abstract

The influence of nitrate and nitrite on growth of Corynebacterium glutamicum under aerobic conditions in shake flasks was analysed. When dissolved oxygen became limiting at higher cell densities, nitrate was reduced almost stoichiometrically to nitrite by nitrate reductase (NarGHJI). The nitrite concentration also declined slowly, presumably as a result of several reactions including reduction to nitric oxide by a side-activity of nitrate reductase. The flavohaemoglobin gene hmp was most strongly upregulated (19-fold) in the presence of nitrite. Hmp is known to catalyse the oxygen-dependent oxidation of nitric oxide to nitrate and, in the absence of oxygen, with a much lower rate the reduction of nitric oxide to nitrous oxide. A Δhmp mutant showed strong growth defects under aerobic conditions in the presence of nitrate, nitrite and the NO-donating reagent sodium nitroprusside, but also under anaerobic nitrate-respiring conditions. Therefore, Hmp is likely to be responsible for nitric oxide conversion to either nitrate or nitrous oxide in C. glutamicum. The results suggest that a cyclic nitrate–nitrite conversion takes place in C. glutamicum under microaerobic conditions.