J. Biochem, 1984, Vol. 95, No. 6 1535-1541
© 1984 Japanese Biochemical Society
research-article |
Redox Properties of Membrane-Bound b-Type Cytochromes and a Soluble c-Type Cytochrome of Nitrate Reductase in a Photodenitrifier, Rhodopseudomonas sphaeroides forma sp. denitrificans
Department of Biology, Tokyo Metropolitan University Fukazawa, Setagaya-ku, Tokyo 158
2To whom correspondence should be addressed.
In order to identify the b-type cytochrome involved in the nitrate reduction in a photodenitrifier, Rhodopseudomonas sphaeroides forma sp. denitrificans, the b-type cytochromes in the spheroplast membranes were characterized. Difference spectra at 77K of spheroplast membranes indicated the presence of two b-type cytochromes with a bands at 556.5 and 562 nm. Three components considered to be of the b-type cytochrome were resolved by anaerobic potentiometric titration at 560–572 nm. Their midpoint potentials at pH 7, Em,7, were –135 mV, +40 mV and + 175 mV and their approximate reduced minus oxidized maxima were determined to be at 565 nm (562 nm at 77K), 560 nm (556.5 nm) and 560 nm (556.5 nm), respectively. These values are almost the same as those reported for R. sphaeroides. The Em,7 value of the cytochrome c involved in the nitrate reductase of this denitrifier was determined to be 250 mV. A b-type cytochrome reduced with NADH and FMN was oxidized by nitrate in chromatophore membranes. The possibility that cytochrome b (Em,7 = 175 mV) is involved in the nitrate reduction is discussed.
1General Research Institute, Mitsui Toatsu Chemicals, Inc., Nishikubo, Chigasaki, Kanagawa 253