PMID:14659546
| Citation |
Raymond, JB, Price, NP and Pavelka, MS Jr (2003) A method for the enzymatic synthesis and HPLC purification of the peptidoglycan precursor UDP-N-acetylmuramic acid. FEMS Microbiol. Lett. 229:83-9 |
|---|---|
| Abstract |
UDP-N-acetylmuramic acid (UDP-MurNAc) is a precursor for peptidoglycan biosynthesis in bacteria. A major difficulty in the study of this pathway is that UDP-MurNAc is not commercially available. We have developed an enzymatic synthesis scheme for UDP-MurNAc using two easily purified Escherichia coli polyhistidine tagged peptidoglycan biosynthesis enzymes, MurZ and MurB, followed by a single-step purification of UDP-MurNAc by high-performance liquid chromatography. The identity of the UDP-MurNAc synthesized by our method was confirmed by electrospray ionization mass spectrometry. Furthermore, we show that the UDP-MurNAc can support a UDP-MurNAc-L-alanine ligase reaction. |
| Links | |
| Keywords |
Alkyl and Aryl Transferases/genetics; Alkyl and Aryl Transferases/metabolism; Carbohydrate Dehydrogenases/genetics; Carbohydrate Dehydrogenases/metabolism; Chromatography, High Pressure Liquid/methods; Escherichia coli; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Enzymologic; Histidine/genetics; Peptide Synthases/genetics; Peptide Synthases/metabolism; Peptidoglycan/biosynthesis; Recombinant Proteins/genetics; Recombinant Proteins/metabolism; Uridine Diphosphate N-Acetylmuramic Acid/analogs & derivatives; Uridine Diphosphate N-Acetylmuramic Acid/biosynthesis; Uridine Diphosphate N-Acetylmuramic Acid/isolation & purification |
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