Pool levels of UDP N-acetylglucosamine and UDP N-acetylglucosamine-enolpyruvate in Escherichia coli and correlation with peptidoglycan synthesis

Abstract

A high-pressure liquid chromatography procedure was developed for the isolation and quantitation of UDP-N-acetylglucosamine, UDP-N-acetylglucosamine-enolpyruvate, and UDP-N-acetylmuramic acid, which are the early cytoplasmic precursors of bacterial peptidoglycan. In exponential-phase cells of E. coli K-12, the intracellular concentration of UDP-N-acetylglucosamine was about 100 μM, whereas that of UDP-n-acetylglucosamine-enolpyruvate was only 2μM. The phosphoenolpyruvate : UDP-N-acetylglucosamine transferase and UDP-N-acetylglucosamine-enolpyruvate reductase activities were investigated in extracts from E. coli. These activities appeared to be present in amounts sufficient for the ongoing rate of peptidoglycan synthesis. Certain uridine nucleotide peptidoglycan precursors were found to inhibit phosphoenolpyruvate : UDP-N-acetylglucosamine transferase activity.