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Chorismate synthase from Staphylococcus aureus

M J Horsburgh1, T J Foster, P T Barth

  • 1Division of Biochemistry & Molecular Biology, University of Glasgow, UK.

Microbiology (Reading, England)
|October 1, 1996
PubMed
Summary
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Researchers cloned and characterized Staphylococcus aureus aroC and ndk genes, revealing gene linkages similar to Bacillus subtilis. Overexpressed chorismate synthase was purified and analyzed, with properties compared to related enzymes.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • The Staphylococcus aureus aroC gene encodes chorismate synthase, essential for folate biosynthesis.
  • The Staphylococcus aureus ndk gene encodes nucleoside diphosphate kinase (Ndk), involved in nucleotide metabolism.
  • Gene organization and function in S. aureus are crucial for understanding bacterial physiology.

Purpose of the Study:

  • To clone and characterize the aroC and ndk genes from Staphylococcus aureus.
  • To investigate the genetic organization and linkage of aroC, ndk, and gerC genes.
  • To overexpress, purify, and characterize Staphylococcus aureus chorismate synthase.

Main Methods:

  • Gene cloning and DNA sequencing of aroC and ndk.
  • Southern hybridization for gene mapping.

Related Experiment Videos

  • Overexpression in Escherichia coli using a T7 promoter.
  • Enzyme purification and characterization (mass spectrometry).

    • Main Results:

    • The aroC gene, encoding a 388-amino acid protein with 61% identity to Bacillus subtilis AroF, was identified.
    • The ndk gene, encoding a 149-amino acid protein highly identical to other bacterial Ndk, was characterized.
    • Linkage of gerC genes with aroC and ndk was observed, similar to B. subtilis.
    • Purified S. aureus chorismate synthase is a homotetramer with a subunit molecular mass of 43024 Da.

    Conclusions:

    • The genetic organization of aroC, ndk, and gerC genes in S. aureus is conserved with B. subtilis.
    • Staphylococcus aureus chorismate synthase has been successfully overexpressed, purified, and characterized.
    • Comparative analysis of S. aureus chorismate synthase properties provides insights into enzyme function across species.