Expression of aspartokinase, dihydrodipicolinic acid synthase and homoserine dehydrogenase during growth of carrot cell suspension cultures on lysine- and threonine-supplemented media.
Reduction in the amounts of activity of the first enzyme, aspartokinase (EC 2.7.2.4) and two branch-point enzymes, dihydrodipicolinic acid synthase (EC 4.2.1.52) and homoserine dehydrogenase (EC 1.1.1.3), located in the pathway for the synthesis of aspartate-family amino acids, occurred when cell suspension cultures of Daucus carota L. var. Danvers were grown in media containing 2 mM threonine or 2 mM lysine, endproducts of the pathway. Activity of the lysine-sensitive form of aspartokinase was decreased when cells were grown in medium containing lysine and the activity of the threonine-sensitive form was decreased when cells were grown in medium containing threonine. Activity of the branch-point enzyme leading to threonine synthesis, homoserine dehydrogenase, was decreased up to 70% in specific activity (units/mg protein) and relative activity (units/g fresh weight) when cells were grown in media containing lysine or threonine. Threonine had no effect on the relative activity of dihydrodipicolinic acid synthase, but decreased its specific activity. Lysine decreased the relative activity of the synthase by up to 40%, but had little effect on its specific activity. The decreased activities of the enzymes were apparently not due to binding of the inhibitory amino acids to the enzymes since homogenization of cells in buffer with 2 mM lysine and threonine did not decrease the measurable enzyme activities. These and other results presented suggest that both forms of the aspartokinase activity and homoserine dehydrogenase activity can be altered by supplementing the growth medium with lysine or threonine.
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis*; D000596:Amino Acids / Q000378:metabolism; D001222:Aspartate Kinase / Q000096:biosynthesis*; D002262:Carboxy-Lyases / Q000096:biosynthesis*; D002478:Cells, Cultured; D006715:Homoserine Dehydrogenase / Q000096:biosynthesis*; D007700:Kinetics; D008239:Lysine / Q000378:metabolism*; D010770:Phosphotransferases / Q000096:biosynthesis*; D010944:Plants / Q000201:enzymology* / Q000378:metabolism; D012459:Salicylates / Q000096:biosynthesis; D013912:Threonine / Q000378:metabolism*
D000596:Amino Acids; D012459:Salicylates; D013912:Threonine; D000429:Alcohol Oxidoreductases; D006715:Homoserine Dehydrogenase; D010770:Phosphotransferases; D001222:Aspartate Kinase; D002262:Carboxy-Lyases; C021586:6-methylsalicylate decarboxylase; D008239:Lysine