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  • 1
    ISSN: 1432-072X
    Keywords: Azotobacter vinelandii ; Ammonium assimilation ; Dinitrogen fixation ; Nitrogenase proteins ; Continuous culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Azotobacter vinelandii was grown at constant growth rate in a chemostat with different molar ratios of sucrose to ammonium (C/N) in the influent media. Both compounds were consumed at essentially the same ratios as were present in the influent media. At low (C/N)-ratios, the cultures were ammonium-limited. At increased (C/N)-ratio ammonium-assimilating cultures additionally began to fix dinitrogen. The (C/N)-ratio at which nitrogenase activity became measurable, increased when the ambient oxygen concentration was increased. Immunoblotting revealed the appearance of nitrogenase proteins when the activity became detectable. Nitrogenase activity as determined either by acetylene reduction or by total nitrogen fixation gave constant relative activities of 1:3.8 (mol of N2 fixed per mol of acetylene reduced) under all sets of conditions used in this investigation. In spite of the oxygen dependent variation of the (C/N)-ratio, nitrogenase became active when the ammonium supply was less than about 14 nmol of ammonium per g of protein. This suggests that oxygen was not directly involved in the onset of dinitrogen fixation.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-072X
    Keywords: Azotobacter vinelandii ; Respiration ; Growth yield coefficient ; Dinitrogen fixation ; Ammonium-assimilation ; Continuous culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Azotobacter vinelandii was grown in continuous culture at constant dilution rate and at different molar ratios of sucrose to ammonium (C/N) in the inflowing medium. The organisms used up essentially all of the carbon and fixed nitrogen sources. Therefore, the (C/N)-ratio in the influent was the same as the (C/N)-ratio of consumption. Starting close to unity, slight increases of the (C/N)-ratio resulted in increases of cellular respiration. Concomitantly, growth yield coefficients on sucrose decreased while the total biomass stayed constant. At there low (C/N)-ratios growth was limited by ammonium with a yield coefficient on ammonium of about 0.07 g protein per mmol of ammonium. Eventually, however, upon furhter increasing the (C/N)-ratio, respiration as well as the yield coefficient on sucrose approached constant values while the biomass levels increased linearly. This result indicated that a transition to sucrose-limited growth had occurred. The (C/N)-ratio, above which respiration and yield coefficients on sucrose approached constancy, increased when the cultures were grown at higher oxygen tension. When the oxygen tension was higher, and at the same (C/N)-ratios, respiratory values increased, and biomass levels as well as yield coefficients decreased. The data suggest control of respiration and thus of growth yield by the ratio of sucrose to ammonium consumed. These observations infer that commencement of dinitrogen fixation kept the internal (C/N)-ratio constant and consequently respiration as well as yield coefficients on sucrose were maintained.
    Type of Medium: Electronic Resource
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