Glutamine synthetase activity in the ruminal bacterium Succinivibrio dextrinosolvens.
Succinivibrio dextrinosolvens C18 was found to possess glutamine synthetase (GS), urease, glutamate dehydrogenase, and several other nitrogen assimilation enzymes. When grown in continuous culture under ammonia limitation, both GS and urease activities were high and glutamate dehydrogenase activity was low, but the opposite activity pattern was observed for growth in the presence of ample ammonia. The addition of high-level (15 mM) ammonium chloride to ammonia-limited cultures resulted in a rapid loss of GS activity as measured by either the gamma-glutamyl transferase or forward assay method with cells or extracts. No similar activity losses occurred for urease, glutamate dehydrogenase, or pyruvate kinase. The GS activity loss was not prevented by the addition of chloramphenicol and rifampin. The GS activity could be recovered by washing or incubating cells in buffer or by the addition of snake venom phosphodiesterase to cell extracts. Manganese inhibited the GS activity (forward assay) of untreated cells but stimulated the GS activity in ammonia-treated cells. Alanine, glycine, and possibly serine were inhibitory to GS activity. Optimal pH values for GS activity were 7.3 and 7.4 for the forward and gamma-glutamyl transferase assays, respectively. The glutamate dehydrogenase activity was NADPH linked and optimal in the presence of KCl. The data are consistent with an adenylylation-deadenylylation control mechanism for GS activity in S. dextrinosolvens, and the GS pathway is a major route for ammonia assimilation under low environmental ammonia levels. The rapid regulation of the ATP-requiring GS activity may be of ecological importance to this strictly anaerobic ruminal bacterium.
Patterson
J A
JA
Hespell
R B
RB
D000641:Ammonia / Q000494:pharmacology; D000818:Animals; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism*; D006089:Gram-Negative Anaerobic Bacteria / Q000187:drug effects / Q000201:enzymology*; D007700:Kinetics; D012417:Rumen / Q000382:microbiology*; D014510:Urease / Q000378:metabolism
D000641:Ammonia; D014510:Urease; D005974:Glutamate-Ammonia Ligase