Enzymatic synthesis and semi-preparative isolation of N-acetylmuramic acid 6-phosphate

Carbohydrate Research
Sandra UnsleberChristoph Mayer

Abstract

N-acetylmuramic acid 6-phosphate (MurNAc-6P) is a constituent of the bacterial peptidoglycan cell wall, serving as an anchor point of secondary cell wall polymers such as teichoic acids, and it is a key metabolite of the peptidoglycan recycling metabolism. Thus, there is a demand for MurNAc-6P as a standard for cell wall compositional and metabolic analyses and, in addition, as a substrate for peptidoglycan recycling enzymes, e.g. MurNAc-6P etherases (MurQ) and MurNAc-6P phosphatases (MupP), or as an effector molecule of transcriptional MurR regulators. However, MurNAc-6P is commercially not available. We report here the facile enzymatic production of MurNAc-6P in mg-scale from MurNAc and ATP, applying Clostridium acetobutylicum kinase MurK, and purification by semi-preparative HPLC. MurNAc-6P was quantified using a coupled enzyme assay, revealing 75-80% overall product yield, and high purity was confirmed by mass spectrometry and proton NMR.

References

Jun 29, 2005·The Journal of Biological Chemistry·Tina JaegerChristoph Mayer
Mar 11, 2008·Nature Reviews. Microbiology·Christopher Weidenmaier, Andreas Peschel
Jun 7, 2008·Microbiology and Molecular Biology Reviews : MMBR·James T Park, Tsuyoshi Uehara
Nov 10, 2009·Bioscience, Biotechnology, and Biochemistry·Daiki IshiiKaneyoshi Yamamoto
Jul 26, 2011·Journal of Bacteriology·Jan Reith, Christoph Mayer
Jul 29, 2011·Applied Microbiology and Biotechnology·Jan Reith, Christoph Mayer
Oct 4, 2011·The EMBO Journal·Yoshikazu KawaiJeff Errington
Jul 9, 2013·Nature Chemical Biology·Jonathan GisinChristoph Mayer
Sep 13, 2013·Annual Review of Microbiology·Stephanie BrownSuzanne Walker
Oct 23, 2016·Journal of Bacteriology·Angela RuscittoAshu Sharma

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Citations

Jul 13, 2019·International Journal of Medical Microbiology : IJMM·Christoph MayerMarina Borisova
Nov 19, 2020·Pharmaceuticals·Fausto QuedaM Manuel B Marques

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Bacterial Cell Wall Structure (ASM)

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.

Bacterial Cell Wall Structure

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.