Regulation of sialic acid metabolism in Escherichia coli: role of N-acylneuraminate pyruvate-lyase.
In Escherichia coli, synthesis of sialic acid is not regulated by allosteric inhibition mediated by cytidine 5'-monophospho-N-acetylneuraminic acid (CMP-NeuNAc). Evidence for the lack of metabolic control by feedback inhibition was demonstrated by measuring the intracellular level of sialic acid and CMP-NeuNAc in mutants defective in sialic acid polymerization and in CMP-NeuNAc synthesis. Polymerization-defective mutants could not synthesize the polysialic acid capsule and accumulated ca. 25-fold more CMP-NeuNAc than the wild type. Mutants unable to activate sialic acid because of a defect in CMP-NeuNAc synthetase accumulated ca. sevenfold more sialic acid than the wild type. An additional threefold increase in sialic acid levels occurred when a mutation resulting in loss of N-acylneuraminate pyruvate-lysase (sialic acid aldolase) was introduced into the CMP-NeuNAc synthetase-deficient mutant. The aldolase mutation could not be introduced into the polymerization-defective mutant, suggesting that any further increase in the intracellular CMP-NeuNAc concentration was toxic. These results show that sialic acid aldolase can regulate the intracellular concentration of sialic acid and therefore the concentration of CMP-NeuNAc. We conclude that regulation of aldolase, mediated by sialic acid induction, is necessary not only for dissimilating sialic acid (E.R. Vimr and F. A. Troy, J. Bacteriol. 164:845-853, 1985) but also for modulating the level of metabolic intermediates in the sialic acid pathway. In agreement with this conclusion, an increase in the intracellular sialic acid concentration was correlated with an increase in aldolase activity. Direct evidence for the central role of aldolase in regulating the metabolic flux of sialic adid in E. coli was provided by the finding that exogenous radiolabeled sialic acid was specifically incorporated into sialyl polymer in aldolase-negative strain but not in the wild type.
D003569:Cytidine Monophosphate N-Acetylneuraminic Acid / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D009154:Mutation; D007652:Oxo-Acid-Lyases / Q000378:metabolism*; D011108:Polymers; D012794:Sialic Acids / Q000378:metabolism*
D011108:Polymers; D012794:Sialic Acids; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid; D007652:Oxo-Acid-Lyases; C018728:N-acetylneuraminate lyase