Truncated hemoglobins in Frankia CcI3: effects of nitrogen source, oxygen concentration, and nitric oxide

Abstract

Frankia strain CcI3 produces 2 truncated hemoglobins, HbN and HbO. Using ion-exchange chromatography, we characterized the expression of the relative amounts of HbN and HbO in −N (nitrogen-fixing) cultures and +N (nitrogen-supplemented) cultures. The −N cultures maintained an approximately constant ratio of HbO to HbN throughout the life of the culture, with HbO constituting 80%–85% of the total hemoglobin produced. In contrast, in +N cultures, HbN was observed to increase over time and HbO decreased. Total hemoglobin as a fraction of total protein was approximately constant throughout the growth phase in −N cultures, while it decreased somewhat in +N cultures. Subjecting −N cultures to a NO generator resulted in increased production of HbN, relative to the controls. Nitrite accumulated in +N cultures, but not in −N cultures. This suggests that the greater amount of HbN in +N cultures might be due to NO produced by the reduction of nitrite. The effects of O2 concentration were determined in +N cultures. Cultures grown in 1% O2 produced about 4 times more HbO than cultures grown in 20% O2. Overall, these results provide evidence for a role of HbN in NO oxidation and for a role of HbO in adaptation to low oxygen concentrations.