Summary
Whole cells of Rhodospirillum rubrum were cultivated in a malate medium lacking bound nitrogen under N2 and tested for their nitrogenase activity by measuring the disappearance of nitrogen manometrically.
Several experimental conditions were relevant in maintaining consistently high activities. These included: a) light intensity, b) substrate concentration, c) concentration of the cell suspension, d) buffer molarity, pH, and e) temperature.
Under our optimal experimental conditions about 6 moles of either l-malate, fumarate, succinate or 10 moles of pyruvate were consumed per 1 mole of molecular nitrogen.
The amount of gas taken up by the cells agreed quantitatively with the increase of bound nitrogen found in the cells by microkjeldahl determinations.
The fixation of molecular nitrogen is suppresed quickly and specifically by very small amounts of ammonia (<5 μg ammonia N/ml). The duration of this inhibition depends on the amount of ammonia available to the cells.
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Schick, H.J. Substrate and light dependent fixation of molecular nitrogen in Rhodospirillum rubrum . Archiv. Mikrobiol. 75, 89–101 (1971). https://doi.org/10.1007/BF00407997
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DOI: https://doi.org/10.1007/BF00407997