Purification and characterization of a nitrous oxide reductase from Thiosphaera pantotropha. Implications for the mechanism of aerobic nitrous oxide reduction.
The aerobic denitrifer Thiosphaera pantotropha is able to reduce simultaneously nitrous oxide and oxygen even after anaerobic growth [Bell, L. C. & Ferguson, S. J. (1991) Biochem J. 273, 423-427]. A nitrous oxide reductase was purified from anaerobically grown T. pantotropha cells. It is argued, on the basis of inhibitor sensitivities and from immunological evidence, that the same nitrous oxide reductase is involved in nitrous oxide reduction in aerobically grown cells. The purified nitrous oxide reductase was shown to have molecular properties very similar to nitrous oxide reductases previously isolated from anaerobically denitrifying bacteria. The visible absorption spectra of the T. pantotropha enzyme resemble those of the oxygen-affected form of nitrous oxide reductases from other organisms. It is thus concluded that the T. pantotropha nitrous oxide reductase is not peculiarly resistant to the structural changes caused by oxygen. The activity of the purified T. pantotropha nitrous oxide reductase was reconstituted in vitro using horse heart cytochrome c, T. pantotropha cytochrome c551 and T. pantotropha pseudoazurin as electron donors. It is suggested on this basis that either of the T. pantotropha electron-carrier proteins are possible physiological electron donors to T. pantotropha nitrous oxide reductase. Oxygen was shown not to inhibit the in-vitro reduction of nitrous oxide with horse heart ferrocytochrome c as electron donor to the reductase.
Department of Biochemistry, University of Oxford, England.
B C
BC
Berks
D
D
Baratta
J
J
Richardson
S J
SJ
Ferguson
D000332:Aerobiosis; D000818:Animals; D002841:Chromatiaceae / Q000201:enzymology*; D003574:Cytochrome c Group / Q000378:metabolism; D006736:Horses; D006863:Hydrogen-Ion Concentration; D009609:Nitrous Oxide / Q000378:metabolism*; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism
D003574:Cytochrome c Group; D010088:Oxidoreductases; C022763:nitrous oxide reductase; D009609:Nitrous Oxide