One-step purification procedure for UDP-N-acetylmuramyl-peptide murein precursors from Bacillus cereus.
A method is described for the rapid isolation of the activated murein precursors UDP-N-acetyl-muramyl-pentapeptide (UDP-MurNAc-pentapeptide) and UDP-MurNAc-tripeptide from Bacillus cereus. After accumulation of the precursors by inhibition of murein synthesis either in the presence of vancomycin (for the pentapeptide precursor) or D-cycloserine (for the tripeptide precursor) the cells were extracted with boiling water. Prior to high pressure liquid chromatography the material was freed from acid precipitable material. UDP-MurNAc-penta- and tripeptide were separated from other components by reversed-phase HPLC on Hypersil ODS using isocratic elution conditions with sodium phosphate buffer. The precursors were obtained with at least 98% purity and a yield of about 50 mumol from a 10-l culture of B. cereus.
Kohlrausch
U
U
Max-Planck-Institut für Entwicklungsbiologie, Abteilung II, Tübingen, F.R.G.
Höltje
J V
JV
D000119:Acetylmuramyl-Alanyl-Isoglutamine / Q000031:analogs & derivatives* / Q000302:isolation & purification; D001409:Bacillus cereus / Q000032:analysis* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D010457:Peptidoglycan / Q000096:biosynthesis*; D011498:Protein Precursors / Q000302:isolation & purification*; D014538:Uridine Diphosphate N-Acetylmuramic Acid / Q000031:analogs & derivatives* / Q000302:isolation & purification; D014640:Vancomycin / Q000494:pharmacology
D010457:Peptidoglycan; D011498:Protein Precursors; D014538:Uridine Diphosphate N-Acetylmuramic Acid; C002225:UDP-N-acetylmuramic acid pentapeptide; D000119:Acetylmuramyl-Alanyl-Isoglutamine; C035104:muramylNAc-Ala-isoGln-Lys-tripeptide; D014640:Vancomycin