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Bacillus subtilis 5'-nucleotidases with various functions and substrate specificities.

Ayako Terakawa1, Ayane Natsume1, Atsushi Okada1

  • 1Department of Agrobioscience, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan.

BMC Microbiology
|October 28, 2016
PubMed
Summary

Bacillus subtilis possesses major 5'-nucleotidases YcsE and YktC, with YktC also aiding oxidative stress resistance. Minor enzymes may function at high substrate levels, and YqeG is crucial for colony formation.

Keywords:
5′-nucleotidaseBacillus subtilisHaloacid dehalogenase superfamilyInositol monophosphataseInositol phosphateNucleoside/nucleotide metabolismOxidative stressPhosphataseProtein motif

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Microbiology

Background:

  • Escherichia coli possesses multiple 5 '-nucleotidases, including NagD, involved in nucleotide hydrolysis.
  • Genes encoding 5 '-nucleotidase in Bacillus subtilis remained largely unidentified.

Purpose of the Study:

  • To identify and characterize 5 '-nucleotidases in Bacillus subtilis.
  • To investigate the roles of identified enzymes in cellular processes and stress responses.

Main Methods:

  • Expression and purification of His6-tagged B. subtilis proteins in E. coli.
  • Enzymatic assays to determine 5 '-nucleotidase and inositol monophosphatase activities.
  • Gene knockout studies to assess the function of identified genes under various conditions.

Main Results:

  • B. subtilis YcsE demonstrated significant 5 '-nucleotidase activity with broad specificity.
  • B. subtilis YktC exhibited both inositol monophosphatase and 5 '-nucleotidase activity (preferential for GMP and IMP).
  • Other identified B. subtilis enzymes (AraL, YutF, YsaA, YqeG) showed weaker nucleotidase activities.
  • Genes ycsE, yktC, and yqeG were induced by oxidative stress.
  • Mutants lacking yktC showed enhanced growth defects under diamide-induced oxidative stress.
  • yqeG was essential for normal colony formation on solid medium.

Conclusions:

  • YcsE and YktC are identified as major 5 '-nucleotidases in B. subtilis.
  • Minor nucleotidases may play a role when substrate concentrations are high.
  • YktC contributes to oxidative stress resistance, and YqeG is important for colony development.