, Volume 16, Issue 2, pp 181–186 | Cite as

Detection and characterisation of catechol 2,3-dioxygenase in an indigenous soil Pseudomonad by MALDI-TOF MS using a column separation

  • Eirini Tsirogianni
  • Michalis Aivaliotis
  • Michael Karas
  • Georgios Tsiotis


The key enzyme catalyzing the second step in the phenol degradation meta-cleavage pathway (C23O) has been purified to homogeneity from a new bacterial strain, which belongs to genus Pseudomonas. The species was growing on phenol as carbon source. The C23O was detected and identified by absorption spectroscopy. The protein was isolated using sucrose density centrifugation and anion exchange chromatography. The purified protein showed a molecular mass of 32 kDa to sodium dodecyl sulfate polyacrylamid gel electrophoresis and an isoelectric point of 5 estimated by analytical isoelectrical focusing. Matrix-assisted laser desorption ionization-time of flight mass spectrometry and peptide mapping was attempted for the identification of the isolated protein and proteins involved in the metabolic pathway.


MALDI; catechol-2,3-dioxygenase extradiol dioxygenase phenol degradation Pseudomonas sp. strain phDV1 



catechol 1,2-dioxygenase


catechol 2,3-dioxygenase


matrix assisted laser desorption ionisation-time of flight


mass spectrometry


sodium dodecyl sulfate polyacrylamid gel electrophoresis


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Copyright information

© Springer 2005

Authors and Affiliations

  • Eirini Tsirogianni
    • 1
  • Michalis Aivaliotis
    • 1
  • Michael Karas
    • 2
  • Georgios Tsiotis
    • 1
  1. 1.Division of Biochemistry, Department of ChemistryUniversity of CreteHeraklionGreece
  2. 2.Institut für Pharmazeutische ChemieInstrumentelle Analytische Chemie Johann Wolfgang Goethe UniversitätFrankfurt am MainGermany

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