1. Enzymatic synthesis and semi-preparative isolation of N-acetylmuramic acid 6-phosphate.
- Author
-
Unsleber S, Borisova M, and Mayer C
- Subjects
- Chemistry Techniques, Synthetic, Clostridium acetobutylicum enzymology, Muramic Acids chemical synthesis, Muramic Acids isolation & purification, Phosphotransferases metabolism
- 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., (Copyright © 2017 Elsevier Ltd. All rights reserved.)
- Published
- 2017
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