MurA or UDP-N-acetylglucosamine enolpyruvyl transferase catalyzes the first committed step in peptidoglycan biosynthesis in bacteria.  It is an essential enzyme and attractive target for anti-bacterial drug discovery. MurA transfers enolpyruvate from phosphoenolpyruvate (PEP) to uridine diphospho-N-acetylglucosamine (UNAG) generating enolpyruvyl-UDPN-acetylglucosamine (EP-UNAG) and inorganic phosphate.

The Bacterial MurA Assay is based on measurement of the inorganic phosphate generated from the MurA reaction.  The inorganic phosphate is detected by light absorbance at 650 nm. The assay reactions and detection can be performed by using 384-well or 96-well assay plates. Alternatively, the assay reaction can be carried out in Eppendorf tubes and the signal is measured using a cuvette. The high throughput assay can be used for screening inhibitors of bacterial MurA in drug discovery research.  It may also be used for characterization of bacterial MurA.

References:                                       

1. Kurnia D. et al, Antibacterial Flavonoids against Oral Bacteria of Enterococcus faecalis ATCC 29212 from Sarang Semut (Myrmecodia pendans) and its Inhibitor Activity against Enzyme MurA. Curr Drug Discov Technol. 2018 Aug 27.
2. Rustemi e. Antimicrobal potencial of actinobacterial inhibitors of mura and murb ligases in bacterium clostridium difficile. University of ljubljana, Thesis (2019).