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.
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Citations
Feb 13, 2018·Frontiers in Microbiology·Isabel HottmannChristoph Mayer
May 8, 2018·The Journal of Biological Chemistry·Mohd Syed AhangarElizabeth Fullam
May 17, 2018·Neural Plasticity·Yong XieBaodong Zheng
Jul 13, 2019·International Journal of Medical Microbiology : IJMM·Christoph MayerMarina Borisova
Nov 19, 2020·Pharmaceuticals·Fausto QuedaM Manuel B Marques
Aug 10, 2017··Christoph MayerMarina Borisova