Uptake of N-acetyl-D-mannosamine: an essential intermediate in polysialic acid biosynthesis by Escherichia coli K92.
The N-acetyl-D-mannosamine (ManNAc) transport system of Escherichia coli K92 was studied when this bacterium was grown in a chemically defined medium containing ManNAc as carbon source. Kinetic measurements were carried out in vivo at 37 degrees C in 25 mM phosphate buffer, pH 7.5. Under these conditions, the uptake rate was linear for at least 15 min and the calculated Km for ManNAc was 280 microM. The transport system was strongly inhibited by sodium arsenate (97%), potassium cyanide (84%) and 2,4-dinitrophenol (88%) added at final concentrations of 1 mM (each). Analysis of bacterial ManNAc phosphotransferase activity revealed in vitro ManNAc phosphorylation activity only when phosphoenolpyruvate was present. These results strongly support the notion that ManNAc uptake depends on a specific phosphotransferase system. Study of specificities showed that N-acetylglucosamine and mannosamine specifically inhibited the transport of ManNAc in this bacterium. Analysis of expression revealed that the ManNAc transport system was induced by ManNAc, glucosamine, galactosamine, mannosamine and mannose but not by N-acetylglucosamine or N-acetylgalactosamine. Moreover, ManNAc permease was subject to glucose repression and cAMP stimulation. Full induction of the ManNAc transport system required the simultaneous presence of both cAMP and ManNAc.
Revilla-Nuin
B
B
Departamento de Bioquímica y Biología Molecular, Universidad de León, Spain.
Reglero
A
A
Ferrero
M A
MA
Rodríguez-Aparicio
L B
LB
D001692:Biological Transport; D004926:Escherichia coli / Q000378:metabolism*; D006595:Hexosamines / Q000378:metabolism*; D012794:Sialic Acids / Q000096:biosynthesis*
D006595:Hexosamines; D012794:Sialic Acids; C021319:polysialic acid; C002022:N-acetylmannosamine