Current Microbiology

, Volume 50, Issue 5, pp 251–256 | Cite as

Identification, Cloning, and Expression of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Gene

  • María J. Massimelli
  • Paola R. Beassoni
  • Marina A. Forrellad
  • José L. Barra
  • Mónica N. Garrido
  • Carlos E. Domenech
  • Angela T. LisaEmail author


Pseudomonas aeruginosa phosphorylcholine phosphatase (PChP) is a periplasmic enzyme produced simultaneously with the hemolytic phospholipase C (PLc-H) when the bacteria are grown in the presence of choline, betaine, dimethylglycine or carnitine. Molecular analysis of the P. aeruginosa mutant JUF8-00, after Tn5-751 mutagenesis, revealed that the PA5292 gene in the P. aeruginosa PAO1 genome was responsible for the synthesis of PChP. The enzyme expressed in E. coli, rPChP-Ec, purified by a chitin-binding column (IMPACT-CN system, New England BioLabs) was homogeneous after SDS-PAGE analysis. PChP was also expressed in P. aeruginosa PAO1-LAC, rPChP-Pa. Both recombinant enzymes exhibited a molecular mass of approximately 40 kDa, as expected for the size of the PA5292 gene, and catalyzed the hydrolysis of phosphorylcholine, phosphorylethanolamine, and p-nitrophenylphosphate. The saturation curve of rPChP-Ec and rPChP-Pa by phosphorylcholine revealed that these recombinant enzymes, like the purified native PChP, also contained the high- and low-affinity sites for phosphorylcholine and that the enzyme activity was inhibited by high substrate concentration.


Recombinant Enzyme Phosphorylcholine Dimethylglycine Periplasmic Extract Phosphorylethanolamine 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank Dr. Fernando J. Irazoqui for advice and help with the preparation of mouse polyclonal anti-rPChP-Ec antibodies, the language consultants Iliana A. Martinez (UNRC) and Ann Lewis (USA) for assistance in the preparation of the manuscript, and Mariela Woelker for technical assistance. C.E.D., A.T.L., M.N.G., and J.L.B. are Career Members of the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). M.J.M., P.R.B., and M.A.F. have a fellowship from CONICET. This work was supported by grants from CONICET, Agencia Córdoba Ciencia and SECYT-UNRC of Argentina.

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Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • María J. Massimelli
    • 1
  • Paola R. Beassoni
    • 1
  • Marina A. Forrellad
    • 1
  • José L. Barra
    • 2
  • Mónica N. Garrido
    • 1
  • Carlos E. Domenech
    • 1
  • Angela T. Lisa
    • 1
    Email author
  1. 1.Departamento de Biología Molecular, Facultad de Ciencias Exactas, Fisicoquímicas y NaturalesUniversidad Nacional de Río CuartoCórdobaArgentina
  2. 2.CIQUIBIC, Departamento de Química BiológicaUniversidad Nacional de CórdobaCórdobaArgentina

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