MurD is a D-Glutamic acid-adding enzyme in the pathway for bacterial cell-wall peptidoglycan synthesis. It is an essential enzyme and attractive target for anti-bacterial drug discovery. MurD catalyses the addition of D-glutamic acid to UDP-MurNAc-L-Ala, generating UDP-MurNAc-dipeptide. The ligation reaction uses ATP hydrolysis as an energy source forming ADP and inorganic phosphate.

The Bacterial MurD Assay is based on measurement of the inorganic phosphate generated from the MurD 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 MurD in drug discovery research.  It may also be used for characterization of bacterial MurD.

E. coli MurD Assay Kit Plus-100 (Catalog No. MURD100KE)

P. aeruginosa MurD Assay Kit Plus-100 (Catalog No. MURD100KP)

S. aureus MurD Assay Kit Plus-100 (Catalog No. MURD100KS)

References:                                       

1. Martinez SR. et al, Identification of the potential biological target of N-benzenesulfonyl-1,2,3,4-tetrahydroquinoline compounds active against gram-positive and gram-negative bacteria. J Biomol Struct Dyn. 23:1-10 (2019).
2. Azam M. A., at al, Structure-based virtual screening to identify inhibitors against Staphylococcus aureus MurD enzyme, Structural Chemistry,  https://doi.org/10.1007/s11224-019-01330-z, April (2019).
3. Jupudi S. et al, Synthesis, molecular docking, binding free energy calculation and molecular dynamics simulation studies of benzothiazol-2-ylcarbamodithioates as Staphylococcus aureus MurD inhibitors. Journal of Receptors and Signal Transduction, Volume 39, Issue 3, (2019).