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A cytochrome c peroxidase from Pseudomonas nautica 617 active at high ionic strength: expression, purification and characterization

Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, ISSN: 0167-4838, Vol: 1434, Issue: 2, Page: 248-259
1999
  • 44
    Citations
  • 0
    Usage
  • 31
    Captures
  • 0
    Mentions
  • 24
    Social Media
Metric Options:   Counts1 Year3 Year

Metrics Details

  • Citations
    44
  • Captures
    31
  • Social Media
    24
    • Shares, Likes & Comments
      24
      • Facebook
        24

Article Description

Cytochrome c peroxidase was expressed in cells of Pseudomonas nautica strain 617 grown under microaerophilic conditions. The 36.5 kDa dihaemic enzyme was purified to electrophoretic homogeneity in three chromatographic steps. N-terminal sequence comparison showed that the Ps. nautica enzyme exhibits a high similarity with the corresponding proteins from Paracoccus denitrificans and Pseudomonas aeruginosa. UV-visible spectra confirm calcium activation of the enzyme through spin state transition of the peroxidatic haem. Monohaemic cytochrome c 552 from Ps. nautica was identified as the physiological electron donor, with a half-saturating concentration of 122 μM and allowing a maximal catalytic centre activity of 116 000 min −1. Using this cytochrome the enzyme retained the same activity even at high ionic strength. There are indications that the interactions between the two redox partners are mainly hydrophobic in nature.

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