Cloning and characterization of Escherichia coli DUF299: a bifunctional ADP-dependent kinase--Pi-dependent pyrophosphorylase from bacteria

BMC Biochem. 2010 Jan 3;11:1. doi: 10.1186/1471-2091-11-1.


Background: Phosphoenolpyruvate synthetase (PEPS; EC catalyzes the synthesis of phosphoenolpyruvate from pyruvate in Escherichia coli when cells are grown on a three carbon source. It also catalyses the anabolic conversion of pyruvate to phosphoenolpyruvate in gluconeogenesis. A bioinformatics search conducted following the successful cloning and expression of maize leaf pyruvate, orthophosphate dikinase regulatory protein (PDRP) revealed the presence of PDRP homologs in more than 300 bacterial species; the PDRP homolog was identified as DUF299.

Results: This paper describes the cloning and expression of both PEPS and DUF299 from E. coli and establishes that E. coli DUF299 catalyzes both the ADP-dependent inactivation and the Pi-dependent activation of PEPS.

Conclusion: This paper represents the first report of a bifunctional regulatory enzyme catalysing an ADP-dependent phosphorylation and a Pi-dependent pyrophosphorylation reaction in bacteria.

MeSH terms

  • Adenosine Diphosphate / metabolism*
  • Cloning, Molecular
  • Escherichia coli / enzymology*
  • Escherichia coli Proteins / classification
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Gluconeogenesis
  • Multienzyme Complexes / classification
  • Multienzyme Complexes / genetics
  • Multienzyme Complexes / metabolism*
  • Phosphoenolpyruvate / metabolism
  • Phosphoric Monoester Hydrolases / classification
  • Phosphoric Monoester Hydrolases / genetics
  • Phosphoric Monoester Hydrolases / metabolism*
  • Phosphotransferases (Paired Acceptors) / classification
  • Phosphotransferases (Paired Acceptors) / genetics
  • Phosphotransferases (Paired Acceptors) / metabolism*
  • Phylogeny
  • Protein Kinases / classification
  • Protein Kinases / genetics
  • Protein Kinases / metabolism*
  • Pyruvate, Orthophosphate Dikinase / classification
  • Pyruvate, Orthophosphate Dikinase / genetics
  • Pyruvate, Orthophosphate Dikinase / metabolism
  • Pyruvic Acid / metabolism
  • Zea mays / enzymology


  • Escherichia coli Proteins
  • Multienzyme Complexes
  • Adenosine Diphosphate
  • Phosphoenolpyruvate
  • Pyruvic Acid
  • Protein Kinases
  • DUF299 protein, E coli
  • Phosphotransferases (Paired Acceptors)
  • Pyruvate, Orthophosphate Dikinase
  • pyruvate, water dikinase
  • Phosphoric Monoester Hydrolases