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Archives of Microbiology

, Volume 165, Issue 1, pp 55–61 | Cite as

Characterization of the membranous denitrification enzymes nitrite reductase (cytochrome cd1) and copper-containing nitrous oxide reductase from Thiobacillus denitrificans

  • U. H. Hole
  • Kai-Uwe Vollack
  • Walter G. Zumft
  • Effi Eisenmann
  • Roman A. Siddiqui
  • Bärbel Friedrich
  • Peter M. H. Kroneck
Original paper

Abstract

Cytochrome cd 1-nitrite reductase and nitrous oxide reductase of Thiobacillus denitrificans were purified and characterized by biochemical and immunochemical methods. In contrast to the generally soluble nature of the denitrification enzymes, these two enzymes were isolated from the membrane fraction of T. denitrificans and remained active after solubilization with Triton X-100. The properties of the membrane-derived enzymes were similar to those of their soluble counterparts from the same organism. Nitrous oxide reductase activity was inhibited by acetylene. Nitrite reductase and nitrous oxide reductase cross-reacted with antisera raised against the soluble enzymes from Pseudomonas stutzeri. The nirS, norBC, and nosZ genes encoding the cytochrome cd 1-nitrite reductase, nitric oxide reductase, and nitrous oxide reductase, respectively, from P. stutzeri hybridized with genomic DNA from T. denitrificans. Cross-reactivity and similar N-terminal amino acid and gene sequences suggest that the primary structures of the Thiobacillus enzymes are homologous to the soluble proteins from P. stutzeri.

Key words Cytochrome cd1 Nitrite reductase Nitrous oxide reductase Denitrification Thiobacillus denitrificans Pseudomonas stutzeri DNA hybridization 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1996

Authors and Affiliations

  • U. H. Hole
    • 1
  • Kai-Uwe Vollack
    • 2
  • Walter G. Zumft
    • 2
  • Effi Eisenmann
    • 3
  • Roman A. Siddiqui
    • 4
  • Bärbel Friedrich
    • 3
  • Peter M. H. Kroneck
    • 1
  1. 1.Fakultät fur Biologie, Universität Konstanz, Postfach 5560, D-78434 Konstanz, Germany Tel. +49-7531-882093; Fax +49-7531-882966DE
  2. 2.Lehrstuhl fur Mikrobiologie, Universität Fridericiana, D-76128 Karlsruhe, GermanyDE
  3. 3.Institut für Biologie, Humboldt-Universität zu Berlin, D-10115 Berlin, GermanyDE
  4. 4.Institut für Pflanzenphysiologie und Mikrobiologie der Freien Universität Berlin, Königin-Luise-Strasse 12–16, D-14195 Berlin, GermanyDE

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