Characterization of a heme c nitrite reductase from a non-ammonifying microorganism, Desulfovibrio vulgaris Hildenborough.
A cytochrome c nitrite reductase (NiR) was purified for the first time from a microorganism not capable of growing on nitrate, the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough. It was isolated from the membranes as a large heterooligomeric complex of 760 kDa, containing two cytochrome c subunits of 56 and 18 kDa. This complex has nitrite and sulfite reductase activities of 685 micromol NH(4)(+)/min/mg and 1.0 micromol H(2)/min/mg. The enzyme was studied by UV-visible and electron paramagnetic resonance (EPR) spectroscopies. The overall redox behavior was determined through a visible redox titration. The data were analyzed with a set of four redox transitions, with an E(0)' of +160 mV (12% of total absorption), -5 mV (38% of total absorption), -110 mV (38% of total absorption) and -210 mV (12% of total absorption) at pH 7.6. The EPR spectra of oxidized and partially reduced NiR show a complex pattern, indicative of multiple heme-heme magnetic interactions. It was found that D. vulgaris Hildenborough is not capable of using nitrite as a terminal electron acceptor. These results indicate that in this organism the NiR is not involved in the dissimilative reduction of nitrite, as is the case with the other similar enzymes isolated so far. The possible role of this enzyme in the detoxification of nitrite and/or in the reduction of sulfite is discussed.
Pereira
I A
IA
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Portugal. ipereira@itqb.unl.pt
LeGall
J
J
Xavier
A V
AV
Teixeira
M
M
D000595:Amino Acid Sequence; D001205:Ascorbic Acid; D003574:Cytochrome c Group / Q000737:chemistry* / Q000302:isolation & purification; D016969:Desulfovibrio vulgaris / Q000201:enzymology* / Q000254:growth & development; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D009572:Nitrite Reductases / Q000737:chemistry*; D010084:Oxidation-Reduction; D012966:Sodium Cyanide; D013053:Spectrophotometry
D003574:Cytochrome c Group; C083951:cytochrome c553; C022302:cytochrome C-552; D009572:Nitrite Reductases; D012966:Sodium Cyanide; D001205:Ascorbic Acid