Cytochrome c2 is essential for electron transfer to nitrous oxide reductase from physiological substrates in Rhodobacter capsulatus and can act as an electron donor to the reductase in vitro Correlation with photoinhibition studies

Richardson, David J. ORCID:, Bell, Louise C., McEwan, Alastair G., Jackson, J. Baz and Ferguson, Stuart J. (1991) Cytochrome c2 is essential for electron transfer to nitrous oxide reductase from physiological substrates in Rhodobacter capsulatus and can act as an electron donor to the reductase in vitro Correlation with photoinhibition studies. European Journal of Biochemistry, 199 (3). pp. 677-683. ISSN 0014-2956

Full text not available from this repository. (Request a copy)


1. Addition of nitrous oxide to a periplasmic fraction released from Rhodobacter capsulatus strains MT1131, N22DNAR+ or AD2 caused oxidation of c‐type cytochrome, as judged by the decrease in absorbance at 550 nm. The periplasmic fraction catalysed reduction of nitrous oxide in the presence of either isoascorbate plus phenazine ethosulphate or reduced methyl viologen. The rates with these two electron donors were similar and were comparable to the activity observed with a quantity of cells equivalent to those from which the periplasm sample had been derived. Activity in the periplasm could not be observed with ascorbate plus 2,3,5,6‐tetramethyl‐p‐phenylenediamine although this reductant was effective with intact cells treated with myxothiazol to block the activity of the cytochrome‐bc1 complex.  2. Cells of R. capsulatus MTG4/S4, a mutant from which the gene for cytochrome c2 has been specifically deleted, did not catalyse detectable rates of nitrous‐oxide reduction. A nitrous‐oxide reductase activity was present, as shown by activity of both cells and a periplasmic fraction with isoascorbate plus phenazine ethosulphate as reductant. The rates in cells and the periplasmic fraction were similar to those observed in the corresponding wild‐type strain (MT1131). In contrast to wild‐type cells, 2,3,5,6‐tetramethyl‐p‐phenylenediamine and N,N,N,N′‐tetramethyl‐p‐phenylenediamine [Ph(NMe2)2] were ineffective as mediators of electrons from isoascorbate. Visible absorption spectra showed that no detectable cytochromes in either the periplasm or intact cells of the MTG4/S4 mutant were oxidised by nitrous oxide.  3. Purified ferroycytochrome c2 from R. capsulatus was oxidised by nitrous oxide in the presence of periplasm from R. capsulatus MTG4/S4. The rate of oxidation was proportional to the amount of periplasm added, but was considerably lower than the rate of nitrous‐oxide reduction observed with the same periplasmic fraction when either ascorbate plus phenazine ethosulphate or reduced methyl viologen were used as substrates. The oxidation of cytochrome c2 was inhibited by acetylene and by low concentrations of NaCl.  4. Oxidation of ferrocytochrome c2 by nitrous oxide was observed when the purified cytochrome was mixed with a preparation of nitrous‐oxide reductase. However, oxidation of ferrocytochrome c′ by nitrous oxide was not observed in the presence of the reductase. The observations with the mutant MTG4/S4 suggest that cytochrome c2 is the only periplasmic cytochrome involved in nitrous‐oxide reduction.  5. Nitrous‐oxide‐dependent oxidation of a c‐type cytochrome was observed in a periplasmic fraction from Paracoccus denitrificans, provided the fraction was first reduced. Nitrous‐oxide‐reductase activity was detected with reduced methyl viologen as electron donor, but not, in contrast to R. capsulatus, with isoascorbate plus phenazine ethosulphate.  6. Inhibition by myxothiazol of nitrous‐oxide reduction by cells of R. capsulatus N22DNAR+ was overcome by isoascorbate plus Ph(NMe2)2. Illumination inhibited the isoascorbate plus Ph(NMe2)2‐dependent activity. In the presence of rotenone [in either the presence or absence of isoascorbate plus Ph(NMe2)2] illumination stimulated the rate of reduction of nitrous oxide. Whereas certain effects of illumination have previously been taken as evidence against a role for cytochrome c2 in nitrous‐oxide respiration, additional observations presented herein lead to reversal of this conclusion.

Item Type: Article
Uncontrolled Keywords: biochemistry ,/dk/atira/pure/subjectarea/asjc/1300/1303
Faculty \ School:
Faculty of Science > School of Biological Sciences
UEA Research Groups: Faculty of Science > Research Groups > Organisms and the Environment
Faculty of Science > Research Groups > Molecular Microbiology
Faculty of Science > Research Centres > Centre for Molecular and Structural Biochemistry
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 15 Jul 2022 13:30
Last Modified: 15 May 2023 00:55
DOI: 10.1111/j.1432-1033.1991.tb16170.x

Actions (login required)

View Item View Item