On the Nutrition of Desulphovibrio desulphuricans

Postgate, J. R.

doi:10.1099/00221287-5-4-714

Cited by 66 publications

(19 citation statements)

References 12 publications

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“…Cytochromes were eluted with starting buffer. After washing with 1 1 10 mM Tris/Cl, pH 8.0, plus 20 mM NaCl, a 20-400 mM NaCl gradient in 10 mM Tris/Cl, pH 8.0, (2 1) was applied to separate the bound proteins into two main fractions: a brownish fraction (60-110 mM NaCl) with the prismane protein (see below), (partially inactivated) periplasmic hydrogenase, adenosine-5'-phosphosulfate (APS) reductase, assimilatory sulfite reductase and a greenish-brown fraction containing desulfoviridin and ferredoxin. The brownish fraction (typically 400 ml) was concentrated using an Amicon device with YM-30 filter.…”

Section: Growth and Isolationmentioning

confidence: 99%

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Purification and biochemical characterization of a putative [6Fe‐6S] prismane‐cluster‐containing protein from Desulfovibrio vulgaris (Hildenborough)

Pierik

Wolbert

Mutsaers

et al. 1992

European Journal of Biochemistry

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A novel iron-sulfur protein has been isolated from the sulfate-reducing bacterium Desulfovibrio vulgaris (Hildenborough). It is a stable monomeric protein, which has a molecular mass of 52 kDa, as determined by sedimentation-equilibrium centrifugation. Analysis of the metal and acid-labile sulfur content of the protein revealed the presence of 6.3 rfr 0.4 Fe/polypeptide and 6.2 & 0.7 S2-/ polypeptide. Non-iron transition metals, heme, fiavin and selenium were absent. Combining these data with the observation of a very anisotropic S = 1/2 [6Fe-6SI3+ prismane-like EPR signal in the dithionite-reduced protein, we believe that we have encountered the first example of a prismanecluster-containing protein. The prismane protein has a slightly acidic amino acid composition and isoelectric point (PI = 4.9). The ultraviolet/visible spectrum is relatively featureless ( E~~~ = 81 mM-' . cm-l, E~~~ = 25 mM-l . cm-', E~~~,~~~ = 14 mM-' . cm-I). The shape of the protein is approximately globular (s20,w = 4.18 S). The N-terminal amino acid sequence is MFSlcFQSic QETAKNTG. Polyclonal antibodies against the protein were raised. Cytoplasmic localization was inferred from subcellular fractionation studies. Cross-reactivity of antibodies against this protein indicated the occurrence of a similar protein in D . vulgaris (Monticello) and Desulfovibrio desulfuricans (ATCC 27774). We have not yet identified a physiological function for the prismane protein despite trials for some relevant enzyme activities.Iron-sulfur clusters constitute the redox-active site of a large number of biological electron-transfer components. The ubiquity of these versatile redox centers is documented by their involvement in vital steps of biochemical pathways: the mitochondria1 respiratory chain, photosynthesis, Krebs' cycle, methanogenesis, nitrogen fixation, sulfate and nitrate reduction [l]. The knowledge of iron-sulfur clusters in biological components in general is based on spectroscopically and crystallographically scrutinized electron-transfer proteins. reactions can be accomplished by iron-sulfur clusters [4]. The best-characterized example is aconitase, for which an X-ray crystallographical structure is available [ 51.Unfortunately, the basic set of structures and their respective spectroscopic properties is not sufficient to explain the EPR characteristics of multielectron redox proteins [6]. Straightforward elucidation of the structure of the iron-sulfur sites is hampered by the lack of sufficient resolution of the crystallographical structures, the absence of amino acid sequence data, and/or the lability of many iron-sulfur centres in the aerobic environment [7, 81. As a working hypothesis for the description of these structures and their magnetism, we have recently proposed the supercluster/superspin concept [9, lo]. This hypothesis explains the absence of classical ironsulfur clusters and the presence of unusual EPR spectra in multielectron redox proteins with a high Fe/S content by proposing the existence of superclusters of more than four ma...

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Section: Growth and Isolationmentioning

confidence: 99%

“…Desuljovihrio vulgaris (Hildenborough) NCIB 8303 was maintained on Postgate's medium [20]. For localization experiments of Desulfovibrio strains, cells were grown in Saunders' N medium [21] to an absorbance of approximately 0.8 at 660 nm.…”

Section: Growth and Isolationmentioning

confidence: 99%

Purification and biochemical characterization of a putative [6Fe‐6S] prismane‐cluster‐containing protein from Desulfovibrio vulgaris (Hildenborough)

Pierik

Wolbert

Mutsaers

et al. 1992

European Journal of Biochemistry

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“…They reduced sulphate, produced desulfoviridin, and were progressively motile vibrios. Although, in nitrogen-fixation experiments, the sulphate-reducing vibrios were the predominant organisms in these isolates, microscopic examination revealed some contaminating bacteria, confirmed by growth on the medium described by Postgate (1951). In older cultures it was sometimes dificult to detect microscopically anything but Desulfovibrio forms ; however, transfer of the culture into fresh medium containing combined nitrogen encouraged growth of the contaminants.…”

Section: Resultsmentioning

confidence: 99%

Nitrogen Fixation by Sulphate-reducing Bacteria

Riederer-Henderson

Wilson

1970

Journal of General Microbiology

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SUMMARYNitrogen fixation has been obained with strains of Desulfovibrio vulgaris and D . gigas, organisms hitherto believed to be incapable of using molecular nitrogen. Fixation has been demonstrated by increases in total nitrogen and by uptake of 15N2. Fixation of N, may be widespread in this genus of the sulphate-reducing bacteria. I N T R O D U C T I O NThe research reported here was begun as a result of Postgate's comment (19653) in a review that 'Most strains of Desulfovibrio do not fix nitrogen'. Enriched cultures from the mud of Lake Mendota, Wisconsin, not completely free of contaminating bacteria, readily fixed N, ; however, the pure cultures intended as negative controls, Desulfovibrio vulgaris NCIB 8303 HILDENBOROUGH and D. gigas NCIB 9332, also fixed in some experiments. Accordingly, the attempt to purify the new isolates was discontinued in favour of a more thorough examination of this unexpected development. The results, reported briefly at the Colloquium on Nitrogen Fixation held at Sanibd Island, Florida (Silver, 1967), are given in more detail here. MATERIALS A N D METHODSCultures. The culture of Desulfovibrio vulgaris strain NCIB 8303 (HILDENBOROUGH) originally was obtained from Sister M. Regina Lanigan, Rosary Hill College (Buffalo, N.Y.); later, subcultures of this same strain were obtained from Professors L. L. Campbell (University of Illinois) and J. R. Postgate (University of Sussex). The purity of the cultures was routinely checked by examination in a phase microscope and inoculation of Postgate's medium A (1951). These subcultures are designated by the letters R, C and P after the strain number 8303. Crude cultures of this organism were isolated from Lake Mendota mud using the technique described by Postgate with 0.02 yo thioglycollate and 0.01 % ascorbic acid added. In the nitrogen fixation * Present address :

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“…The sulfate reducer D. vulgaris (Hildenborough) NCIB 8303 was maintained on Postgate's medium [14] ; large-scale growth at 35 "C on lactate/sulfate was in modified Saunders' medium [15]. Stirred batch cultures of 240 1 were grown under nitrogen to an A,,, value of approximately 0.8 in 60 h from a 1 % inoculum in a home-built anaerobic glass fermentor.…”

Section: Organism Growth and Isolationmentioning

confidence: 99%

“…Subsequently, however, the structural gene for Hmc was sequenced by Pollock et al, who found 16 regions encoding Cys-Xaa-Xaa-Cys-His motifs for the potential covalent binding of 16 hemes [4]. Furthermore, with this gene transferred to Desulfovibrio desulfuricans, strain G200, an overexpression system was constructed and the purified recombinant Hmc contained [13][14][15][16] hemes by chemical analysis [lo].…”

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confidence: 99%

Axial Coordination and Reduction Potentials of the Sixteen Hemes in High‐Molecular‐Mass Cytochrome c from Desulfovibrio Vulgaris (Hildenborough)

Verhagen

Pierik

Wolbert

et al. 1994

European Journal of Biochemistry

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A spectroelectrochemical study is described of the sixteen hemes in the high-molecular-mass, monomeric cytochrome c (Hmc) from the periplasmic space of Desulfovibrio vulgaris, strain Hildenborough. One of the hemes has special properties. In the oxidized state at pH 7 it is predominantly high-spin, S = 512, with a gi value of less than 6 indicative of quantum-mechanical mixing with a low-lying (800 cm-') S = 312 state; the balance is probably a low-spin derivative. The highspin heme has an E,,,7.5 value of +61 mV. The fifteen other hemes are low-spin bis-histidine coordinated with Em,7.5 values of approximately -0.20 V. Two of these hemes exhibit very anisotropic EPR spectra with a g, value of 3.65 characteristic for strained bis-histidine coordination. A previous proposal, namely that methionine is coordinated to one of the hemes Sulfate reducers in general have a high heme content. The heme porphyrin structure is used by these bacteria in two ways. Sirohydrochlorin, the basic component of siroheme, is the prosthetic group of dissimilatory and assimilatory sulfite reductases in the cytoplasm. Protoporphyrin IX is the basic component for b-type and c-type cytochromes in the periplasm, in the cytoplasmic membrane, and also possibly in the cytoplasm. Although some of these proteins have been studied in considerable detail, our knowledge of the number, type, location, biosynthesis, and, especially, the function of these cytochromes is incomplete [l].For example, in the intensively studied Desulfovibrio vulgaris, strain Hildenborough, no cytochromes have yet been identified in the cytoplasm or in the cytoplasmic membrane. Export to the periplasm has been found for three c-type cytochromes, monoheme cSs3 [2], tetraheme c3 [3], and multiheme high-molecular-mass cytochrome c (Hmc) [4] ; the functions of these cytochromes are either debatable or unknown. The existence of a proposed fourth cytochrome, cytochrome c3 (26 kDa) [ 5 ] , has been questioned [4].

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On the Nutrition of Desulphovibrio desulphuricans

Cited by 66 publications

References 12 publications

Purification and biochemical characterization of a putative [6Fe‐6S] prismane‐cluster‐containing protein from Desulfovibrio vulgaris (Hildenborough)

Purification and biochemical characterization of a putative [6Fe‐6S] prismane‐cluster‐containing protein from Desulfovibrio vulgaris (Hildenborough)

Nitrogen Fixation by Sulphate-reducing Bacteria

Axial Coordination and Reduction Potentials of the Sixteen Hemes in High‐Molecular‐Mass Cytochrome c from Desulfovibrio Vulgaris (Hildenborough)

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