The effect of various culture conditions on the levels of ammonia assimilatory enzymes of Corynebacterium callunae.
Corynebacterium callunae (NCIB 10338) grows faster on glutamate than ammonia when used as sole nitrogen sources. The levels of glutamine synthetase (GS; EC 6.3.1.2) and glutamate synthase (GOGAT; EC 1.4.1.13) of C. callunae were found to be influenced by the nitrogen source. Accordingly, the levels of GS and GOGAT activities were decreased markedly under conditions of ammonia excess and increased under low nitrogen conditions. In contrast, glutamate dehydrogenase (GDH; EC 1.4.1.4) activities were not significantly affected by the type or the concentration of the nitrogen source supplied. The carbon source in the growth medium could also affect GDH, GS and GOGAT levels. Of the carbon sources tested in the presence of 2 mM or 10 mM ammonium chloride as the nitrogen source pyruvate, acetate, fumarate and malate caused a decrease in the levels of all three enzymes as compared with glucose. GDH, GS and GOGAT levels were slightly influenced by aeration. Also, the enzyme levels varied with the growth phase. Methionine sulfoximine, an analogue of glutamine, markedly inhibited both the growth of C. callunae cells and the transferase activity of GS. The apparent Km values of GDH for ammonia and glutamate were 17.2 mM and 69.1 mM, respectively. In the NADPH-dependent reaction of GOGAT, the apparent Km values were 0.1 mM for alpha-ketoglutarate and 0.22 mM for glutamine.
Department of Biology, University of Istanbul, Faculty of Science, Vezneciler, Turkey.
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D000641:Ammonia / Q000378:metabolism*; D003352:Corynebacterium / Q000201:enzymology* / Q000254:growth & development; D003470:Culture Media; D005650:Fumarates / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D005969:Glutamate Dehydrogenase / Q000032:analysis*; D005970:Glutamate Synthase / Q000032:analysis*; D005974:Glutamate-Ammonia Ligase / Q000032:analysis* / Q000037:antagonists & inhibitors; D005971:Glutamates / Q000096:biosynthesis / Q000378:metabolism; D018698:Glutamic Acid; D007700:Kinetics; D008293:Malates / Q000378:metabolism; D008717:Methionine Sulfoximine / Q000494:pharmacology
D003470:Culture Media; D005650:Fumarates; D005971:Glutamates; D008293:Malates; D008717:Methionine Sulfoximine; D018698:Glutamic Acid; D000641:Ammonia; C030298:malic acid; C032005:fumaric acid; D005970:Glutamate Synthase; D005969:Glutamate Dehydrogenase; D005974:Glutamate-Ammonia Ligase; D005947:Glucose