The nitrate- and nitrite-dependent O2-evolution in N2 by Ankistrodesmus braunii

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Abstract
O2-evolution in N2 by young cells of synchronous Ankistrodesmus braunii was measured manometrically in the presence and in the absence of nitrate or nitrite. Nitrate-starved cells produce O2 as a function of nitrate concentration with an optimum at 10 mM nitrate. The optimum rates are strongly dependent upon the pH of the medium culminating at pH 8.0. Nitrite excretion is initially slow and has its optimum at the same nitrate concentration as O2-evolution. It is slower under anaerobic conditions than in the presence of CO2 and slower at pH 5.6 than at pH 8.0. At pH 8.0 in N2 an accumulation of nitrite starts 40 to 100 min after the addition of nitrate to the algae. O2-evolution is faster with nitrite than with nitrate. No optimum curve is observed at pH 8.0 with various concentrations of nitrite in the medium; however, at pH 5.6 a distinct peak is found at 3 mM nitrite. This peak is found for a fast short-time reaction as well as for the following low rates of O2-evolution. The uncoupler carbonylcyanide-m-chlorophenylhydrazone (CCCP) equally inhibits nitrate- and nitrite-dependent O2-evolution and the incorporation of 32P into cellular phosphate compounds. There is no indication of uncoupling in vivo of nitrite reduction which is completely independent of ATP in vitro. The inhibition of the nitrate-dependent O2-evolution by high concentrations of nitrate cannot be explained by an accumulation of products such as ammonia or nitrite.