One-step purification procedure for UDP-N-acetylmuramyl-peptide murein precursors from Bacillus cereus

1 March 1991

journal article
Published by Oxford University Press (OUP) in FEMS Microbiology Letters

Vol. 78 (2-3) , 253-257
https://doi.org/10.1016/0378-1097(91)90166-8

Abstract

A method is described for the rapid isolation of the activated murein precursors UDP-N-acetylmuramyl-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 μmol from a 10–1 culture of B. cereus.

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