Expression of peptidylarginine deiminase from Porphyromonas gingivalis in Escherichia coli: enzyme purification and characterization

Arch Biochem Biophys. 2009 Aug 1;488(1):14-22. doi: 10.1016/ Epub 2009 Jun 21.


Porphyromonas gingivalis peptidylarginine deiminase (PAD) catalyzes the deimination of peptidylarginine residues of various peptides to produce peptidylcitrulline and ammonia. P. gingivalis is associated with adult-onset periodontitis and cardiovascular disease, and its proliferation depends on secretion of PAD. We have expressed two recombinant forms of the P. gingivalis PAD in Escherichia coli, a truncated form with a 43-amino acid N-terminal deletion and the full-length form of PAD as predicted from the DNA sequence. Both forms contain a poly-His tag and Xpress epitope at the N-terminus to aid in detection and purification. The activities and stabilities of these two forms have been evaluated. PAD is cold sensitive; it aggregates within 30 min at 4 degrees C, and optimal storage conditions are at 25 degrees C in the presence of a reducing agent. PAD is not a metalloenzyme and does not need a cofactor for catalysis or stability. Multiple l-arginine analogs, various arginine-containing peptides, and free l-arginine were used to evaluate substrate specificity and determine kinetic parameters.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Biocatalysis
  • Coenzymes / metabolism
  • Enzyme Stability
  • Escherichia coli / genetics*
  • Gene Deletion
  • Gene Expression
  • Hydrogen-Ion Concentration
  • Hydrolases / biosynthesis
  • Hydrolases / chemistry
  • Hydrolases / isolation & purification*
  • Hydrolases / metabolism*
  • Imines / metabolism
  • Metals / metabolism
  • Molecular Sequence Data
  • Molecular Weight
  • Porphyromonas gingivalis / enzymology*
  • Porphyromonas gingivalis / genetics
  • Protein-Arginine Deiminases
  • Sequence Analysis, DNA
  • Substrate Specificity


  • Coenzymes
  • Imines
  • Metals
  • Hydrolases
  • Protein-Arginine Deiminases