Abstract
A procedure was devised for analyzing in vivo nitrogenase activity in Beggiatoa alba B18LD which involves: (1) the induction of nitrogenase in cells pre-grown on NH4Cl, by washing the cells free of NH4Cl and lowering their exposure to oxygen, and (2) measuring acetylene reduction by these cells. Using this induction methodology we examined the effects of pH, temperature, and nitrogenous compounds on in vivo nitrogenase induction and activity in Beggiatoa alba B18LD. Nitrate and nitrite repressed the induction of nitrogenase activity, but glutamine did not. Induction and activity had a combined pH optimum of 6.5 to 8.0, and activity had a temperature optimum of 29°C. Ammonium and urea caused immediate inhibition of nitrogenase activity, but nitrate, nitrite, glutamine, asparagine, and other amino acids did not. Ammonium-induced inhibition was transient and incomplete, and the duration of inhibition increased in direct proportion to the amount of ammonium added. Methionine sulfoximine, a glutamine synthetase inhibitor, at a final concentration of 50 μM blocked ammonium uptake by cells, but did not prevent nitrogenase inhibition if added before ammonium. Our results imply that B. alba nitrogenase inhibition by ammonium: (1) is not directly caused by ammonium assimilation products, (2) is probably not due to an enzymatic inactivation, and (3) may be related to ammonium transport.
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Abbreviations
- MSX:
-
methionine sulfoximine
- MES:
-
2[N-morpholino]ethanesulfonic acid
- bis Tris propane:
-
1,3-bis[tris-(hydroxymethyl)-methylamino]propane
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Approved for publication by the director of the Louisiana Agricultural Experiment Station as manuscript no. 87-33-1305
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Polman, J.K., Larkin, J.M. Properties of in vivo nitrogenase activity in Beggiatoa alba . Arch. Microbiol. 150, 126–130 (1988). https://doi.org/10.1007/BF00425151
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DOI: https://doi.org/10.1007/BF00425151