Cloning and expression of Staphylococcus aureus and Streptococcus pyogenes murD genes encoding uridine diphosphate N-acetylmuramoyl-L-alanine:D-glutamate ligases

Mohamed El-Sherbeini, Wayne M. Geissler, Jamya Pittman, Xiling Yuan, Kenny K. Wong, David L. Pompliano

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

Bacterial UDP-N-acetylmuramyl-L-alanine:D-glutamate ligase (MurD), a cytoplasmic peptidoglycan biosynthetic enzyme, catalyzes the ATP-dependent addition of D-glutamate to an alanyl residue of the UDP-N-acetylmuramyl-L-alanine precursor, generating the dipeptide. The murD gene was cloned from both Staphylococcus aureus and Streptococcus pyogenes. Sequence analysis of the S. aureus murD gene revealed an open reading frame of 449 amino acids. The deduced aa sequence of S. aureus MurD is highly homologous to MurD from Escherichia coli, Haemophilus influenzae, Bacillus subtilis and St. pyogenes. Recombinant MurD protein from both S. aureus and St. pyogenes was separately overproduced in E. coli and purified as His-tagged fusion. Both recombinant enzymes catalyzed the ATP-dependent addition of D-glutamate to the precursor sugar peptide.

Original languageEnglish
Pages (from-to)117-125
Number of pages9
JournalGene
Volume210
Issue number1
DOIs
StatePublished - 27 Mar 1998

Keywords

  • Bacterial cell wall
  • Gram-positive cocci
  • Peptidoglycan

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