Characterization of the membranous denitrification enzymes nitrite reductase (cytochrome cd1) and copper-containing nitrous oxide reductase from Thiobacillus denitrificans |
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Authors: | U H Hole Kai-Uwe Vollack Walter G Zumft Effi Eisenmann Roman A Siddiqui Bärbel Friedrich Peter M H Kroneck |
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Institution: | Fakult?t fur Biologie, Universit?t Konstanz, Postfach 5560, D-78434 Konstanz, Germany Tel. +49-7531-882093; Fax +49-7531-882966, DE Lehrstuhl fur Mikrobiologie, Universit?t Fridericiana, D-76128 Karlsruhe, Germany, DE Institut für Biologie, Humboldt-Universit?t zu Berlin, D-10115 Berlin, Germany, DE Institut für Pflanzenphysiologie und Mikrobiologie der Freien Universit?t Berlin, K?nigin-Luise-Strasse 12–16, D-14195 Berlin, Germany, DE
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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.
Received: 18 August 1995 / Accepted: 30 October 1995 |
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Keywords: | Cytochrome cd1 Nitrite reductase Nitrous oxide reductase Denitrification Thiobacillus denitrificans Pseudomonas stutzeri DNA hybridization |
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