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reduction of uranium by cytochrome c3 of desulfovibrio vulgaris.the mechanism for u(vi) reduction by desulfovibrio vulgaris (hildenborough) was investigated. the h2-dependent u(vi) reductase activity in the soluble fraction of the cells was lost when the soluble fraction was passed over a cationic exchange column which extracted cytochrome c3. addition of cytochrome c3 back to the soluble fraction that had been passed over the cationic exchange column restored the u(vi)-reducing capacity. reduced cytochrome c3 was oxidized by u(vi), as was a c-type cytochrom ...19938285665
dcra, a c-type heme-containing methyl-accepting protein from desulfovibrio vulgaris hildenborough, senses the oxygen concentration or redox potential of the environment.the amino acid sequence of dcra from desulfovibrio vulgaris hildenborough, a strictly anaerobic, sulfate-reducing bacterium, indicated homology with the methyl-accepting chemotaxis proteins from enteric bacteria (a. dolla, r. fu, m. j. brumlik, and g. voordouw, j. bacteriol. 174:1726-1733, 1992). the homology is restricted to the cytoplasmic c-terminal signaling domain. the periplasmic n-terminal sensor domain was found to contain a unique sequence, chhch, corresponding to a consensus c-type hem ...19948288528
identification of a large family of genes for putative chemoreceptor proteins in an ordered library of the desulfovibrio vulgaris hildenborough genome.a library of 879 recombinant lambda phages, constructed for the genome of desulfovibrio vulgaris hildenborough, has been ordered by restriction fingerprinting. restriction endonuclease hinfi digestion patterns were entered into a data base and sorted into 87 overlapping groups (contigs), with 19 clones remaining unattached. eight of ten cloned genes of d. vulgaris, including dcra, which encodes a transmembrane methyl-accepting protein, were assigned to contigs. probing of a filter containing the ...19948288529
characterization of a small plasmid from desulfovibrio desulfuricans and its use for shuttle vector construction.a 2.3-kb plasmid present in about 20 copies per genome was identified in extracts of desulfovibrio desulfuricans g100a and designated pbg1. it appears to be unable to replicate in escherichia coli. although composite plasmids of pbg1 inserted into ptz18u are stable in e. coli, few if any pbg1-specific transcripts are detectable. the plasmid sequence reveals several features typical of the origin of replication of non-cole1 enterobacterial plasmids as well as several potential open reading frames ...19938320227
microbial reduction of sulfur dioxide with pretreated sewage sludge and elemental hydrogen as electron donors.it has been demonstrated tht heat- and alkali-pretreated-sewage sludge may serve as an electron donor and carbon source for so2 reduction by desulfovibrio desulfuricans. a continuous d. desulfuricans culture was operated for 6 mo with complete reduction of so2 to h2s. the culture required only minor amounts of mineral nutrients in addition to pretreated sewage sludge. it has also been shown that the sulfate-reducing bacterium desulfotomaculum orientis can be grown on h2 as an energy source, co2 ...19938323272
effects of thiols and mercurials on the periplasmic hydrogenase from desulfovibrio vulgaris (hildenborough).the h2-oxidation, h2-production and h-3h-exchange activities of the periplasmic hydrogenase from desulfovibrio vulgaris (hildenborough) were almost completely abolished by hg(ii) and the organic mercurials p-chloromercuribenzoate (pcmb) and p-hydroxymercuriphenylsulphonate. the thiol-modifying reagents n-ethylmaleimide, iodoacetate, dithionitrobenzoate and 2-nitro-5-thiocyanobenzoate had no effect on the activities. kinetic and spectroscopic measurements suggest that inactivation by pcmb involve ...19938328964
the hmc operon of desulfovibrio vulgaris subsp. vulgaris hildenborough encodes a potential transmembrane redox protein complex.the nucleotide sequence of the hmc operon from desulfovibrio vulgaris subsp. vulgaris hildenborough indicated the presence of eight open reading frames, encoding proteins orf1 to orf6, rrf1, and rrf2. orf1 is the periplasmic, high-molecular-weight cytochrome (hmc) containing 16 c-type hemes and described before (w. b. r. pollock, m. loutfi, m. bruschi, b. j. rapp-giles, j. d. wall, and g. voordouw, j. bacteriol. 173:220-228, 1991). orf2 is a transmembrane redox protein with four iron-sulfur clus ...19938335628
ileal symbiont intracellularis, an obligate intracellular bacterium of porcine intestines showing a relationship to desulfovibrio species.a new genus and species of obligate intracellular-bacteria found in porcine intestines are described. growth on any bacteriological medium deprived of living cells has not been demonstrated. the organism has been grown intracellularly in cell culture. the 16s rrna gene sequence data, dna probe results, and microscopic observations provide evidence that these bacteria differ from those in other described genera and that they belong to the delta subdivision of the class proteobacteria. we have amp ...19938347512
subunit composition, crystallization and preliminary crystallographic studies of the desulfovibrio gigas aldehyde oxidoreductase containing molybdenum and [2fe-2s] centers.the desulfovibrio gigas aldehyde oxidoreductase contains molybdenum bound to a pterin cofactor and [2fe-2s] centers. the enzyme was characterized by sds/page, gel-filtration and analytical ultracentrifugation experiments. it was crystallized at 4 degrees c, ph 7.2, using isopropanol and mgcl2 as precipitants. the crystals diffract beyond 0.3-nm (3.0-a) resolution and belong to space group p6(1)22 or its enantiomorph, with cell dimensions a = b = 14.45 nm and c = 16.32 nm. there is one subunit/as ...19938354279
microbial transformation of nitroaromatic compounds under anaerobic conditions.the transformation of several mono- and dinitroaromatic compounds (tested at 50-200 microm) by methanogenic bacteria, sulphate-reducing bacteria and clostridia was studied. some of the nitroaromatics tested were transformed chemically by 1.5 mm quantities of culture media reducing agents, like cysteine or sulphide. this abiotic reduction occurred at the o-nitro-groups preferentially. nitrophenols, p-nitroaniline and p-nitrobenzoic acid were completely transformed biologically into the correspond ...19938360625
chromophores of dissimilatory sulphite reductase from wild strains desulfovibrio desulfuricans. 19938372578
aerobic metabolism of carbon reserves by the "obligate anaerobe" desulfovibrio gigas.the sulfate-reducing bacterium, desulfovibrio gigas, is shown by in vivo 31p-nmr to be capable of generating ntp from the utilization of internal carbon reserves both in anaerobic and in aerobic conditions. acetate, glycerol and ethanol are the major end-products, but the production of alcohols decreases strongly when oxygen is present. when the glycolytic pathway is inhibited with fluoride, ntp levels decrease drastically but can be remarkably restored when an electron acceptor, such as oxygen, ...19938373395
purification and characterization of an nadh-rubredoxin oxidoreductase involved in the utilization of oxygen by desulfovibrio gigas.an nadh--rubredoxin oxidoreductase previously isolated from desulfovibrio gigas [legall, j. (1968) ann. inst. pasteur 114, 109-115] has now been fully purified and further characterized. it contains two subunits of 27 kda and 32 kda. with two mid-point redox potentials of -295 mv and -325 mv, this fmn- and fad-containing protein can induce the specific reduction of d. gigas rubredoxin. in contrast, rubredoxins from the other desulfovibrio species or desulforedoxin from d. gigas show very low rea ...19938375383
nigerythrin and rubrerythrin from desulfovibrio vulgaris each contain two mononuclear iron centers and two dinuclear iron clusters.the trivial name 'rubr-erythrin' is a contraction of two other trivial names: rubredoxin (ruber, red) and hemerythrin. it names a protein of undetermined biological function which putatively carries rubredoxin-like mononuclear iron and hemerythrin-like dinuclear iron. the name 'nigerythrin' (niger, black) is an analogy of rubrerythrin. it identifies a second protein of undetermined function which has prosthetic groups similar to rubrerythrin. rubrerythrin was initially described [legall, j., pri ...19938383040
electrochemical studies of the hexaheme nitrite reductase from desulfovibrio desulfuricans atcc 27774.the electron-transfer kinetics between three different mediators and the hexahemic enzyme nitrite reductase isolated from desulfovibrio desulfuricans (atcc 27774) were investigated by cyclic voltammetry and by chronoamperometry. the mediators, methyl viologen, desulfovibrio vulgaris (hildenborough) cytochrome c3 and d. desulfuricans (atcc 27774) cytochrome c3 differ in structure, redox potential and charge. the reduced form of each mediator exchanged electrons with nitrite reductase. second-orde ...19938383043
amino-acid sequence of cytochrome c-553 from desulfovibrio desulfuricans norway.the amino-acid sequence of the cytochrome c-553 from desulfovibrio desulfuricans norway has been determined and compared with that of two different cytochromes c-553 from d. vulgaris already described and with that cytochrome c-551 from pseudomonas. this low-molecular-weight monohemic cytochrome comprises 80 amino acids and has the typical characteristics of small cytochromes such as mitochondrial cytochromes. secondary-structure predictions are deduced from sequence data and are compared with x ...19938383535
isolation and characterization of flavoredoxin, a new flavoprotein that permits in vitro reconstitution of an electron transfer chain from molecular hydrogen to sulfite reduction in the bacterium desulfovibrio gigas.a new fmn-containing flavoprotein isolated from desulfovibrio gigas provided maximum coupling efficiency for the reduction of bisulfite from molecular h2. this protein, which is distinct from flavodoxin and for which the name flavoredoxin is proposed, is required for reconstitution of an electron transfer chain between hydrogenase and bisulfite reductase. a ca(2+)-binding protein functions as a modulator in the presence of ca2+ in the process. the finding of a membrane-bound cytochrome c with a ...19938387752
rapid comparison of the cytochrome c3 gene from nine strains of desulfovibrio vulgaris using polymerase chain reaction amplification.polymerase chain reaction amplification was used to compare different regions of the cytochrome c3 gene from nine strains of desulfovibrio vulgaris, to examine homology within the species. six 30-base polymerase chain reaction primers and three probes were synthesized on the basis of the published nucleic acid sequence of the cytochrome c3 gene from d. vulgaris, ncimb 8303. amplifications were performed on genomic dna isolated from ncimb 8303 as well as eight other strains. six strains, ncimb 83 ...19938388770
electron transport components of the parasitic protozoon giardia lamblia.the energy metabolism of the intestinal parasite, giardia lamblia, involves the iron-sulphur protein, pyruvate:ferredoxin oxidoreductase. cell fractionation studies showed that this enzyme is associated with the membranes. nadh and nadph dehydrogenases were found in both the membrane and cytosolic fractions. epr spectroscopic studies showed the presence of iron-sulphur clusters in the membrane fraction and in the cytosolic fraction, non-sedimentable at 6 x 10(6) g.min. an acidic, soluble protein ...19938391475
spectroscopic characterization of 57fe-reconstituted rubrerythrin, a non-heme iron protein with structural analogies to ribonucleotide reductase.rubrerythrin, a contraction of rubredoxin and hemerythrin, is the trivial name given to a non-heme iron protein isolated from desulfovibrio vulgaris (hildenborough). this protein, whose physiological function is unknown, was first characterized by j. legall et al. [(1988) biochemistry 28, 1636] as being a homodimer of subunit m(r) = 21,900 with four fe per homodimer distributed as two rubredoxin-type fes4 centers and one hemerythrin-type diiron cluster. subsequent analysis of the amino acid sequ ...19938395205
protein conformational changes in tetraheme cytochromes detected by ftir spectroelectrochemistry: desulfovibrio desulfuricans norway 4 and desulfovibrio gigas cytochromes c3.the conformational change coupled to the redox processes of two tetraheme cytochromes c3 from bacteria of the genus desulfovibrio have been studied by uv-vis and ftir difference spectroscopy combined with protein electrochemistry. two pairs of equivalent hemes were found in desulfovibrio desulfuricans norway 4 cytochrome c3 by uv-vis spectroelectrochemical redox titration in an optically transparent thin-layer electrochemical cell. in contrast to this, desulfovibrio gigas cytochrome c3 showed a ...19938396427
intramolecular electron transfer in ferredoxin ii from desulfovibrio desulfuricans norway.in order to elucidate the role of the two (4fe-4s) clusters in ferredoxins and to determine whether an electron-transfer mechanism may occur between the clusters, the in vitro reduction of cytochrome c3 and cytochrome c553 by desulfovibrio desulfuricans norway ferredoxin ii was studied using spectrophotometric techniques. ferredoxin ii, covalently cross-linked with either cytochrome c3 or c553, is an obligate intermediate in cytochrome reduction by pyruvate dehydrogenase. both titration of the c ...19938396440
characterization of the structure and redox behaviour of cytochrome c3 from desulfovibrio baculatus by 1h-nuclear-magnetic-resonance spectroscopy.complete assignment of the aromatic and haem proton resonances in the cytochromes c3 isolated from desulfovibrio baculatus strains (norway 4, dsm 1741) and (dsm 1743) was achieved using one- and two-dimensional 1h n.m.r. nuclear overhauser enhancements observed between haem and aromatic resonances and between resonances due to different haems, together with the ring-current contributions to the chemical shifts of haem resonances, support the argument that the haem core architecture is conserved ...19938397513
structural studies on desulfovibrio gigas cytochrome c3 by two-dimensional 1h-nuclear-magnetic-resonance spectroscopy.several aromatic amino acid residues and haem resonances in the fully reduced form of desulfovibrio gigas cytochrome c3 are assigned, using two-dimensional 1h n.m.r., on the basis of the interactions between the protons of the aromatic amino acids and the haem protons as well as the intrahaem distances known from the x-ray structure [kissinger (1989) ph.d. thesis, washington state university]. the interhaem interactions observed in the n.m.r. spectra are in full agreement with the d. gigas x-ray ...19938397514
the dissimilatory sulfite reductase from desulfosarcina variabilis is a desulforubidin containing uncoupled metalated sirohemes and s = 9/2 iron-sulfur clusters.the active site of escherichia coli nadph-sulfite reductase has previously been modeled as a siroheme with its iron bridged to a nearby iron-sulfur cubane, resulting in antiferromagnetic exchange coupling between all iron atoms. the model has been suggested to hold also for other sulfite reductases and nitrite reductases. we have recently challenged the generality of the model with the finding that the epr of fe/s in dissimilatory sulfite reductase (desulfoviridin) from desulfovibrio vulgaris in ...19938399175
characterization of d. desulfuricans (atcc 27774) [nife] hydrogenase epr and redox properties of the native and the dihydrogen reacted states.redox intermediates of d. desulfuricans atcc 27774 [nife] hydrogenase were generated under dihydrogen. detailed redox titrations, coupled to epr measurements, give access to the mid-point redox potentials of the iron-sulfur centers and of the nickel-b signal that represents the ready form of the enzyme. the interaction between the dihydrogen molecule and the nickel centre was probed by the observation of an isotopic effect on the epr signals detected in turnover conditions, by comparison of the ...19938399280
environmental scanning electron microscope (esem) evaluation of crystal and plaque formation associated with biocorrosion.the biofilm attributed to desulfovibrio vulgaris growing in the presence of ferrous metals was examined with an environmental scanning electron microscope. this novel microscope produced images of iron sulfide colloids and other iron containing structures that had not been reported previously. a plaque composed of iron sulfide enveloped the surface of the corroding metal while crystals containing magnesium, iron, sulfur, and phosphorus were present in the culture where corrosion was in progress. ...19938400436
carboxy-terminal processing of the large subunit of [nife] hydrogenases.two electrophoretic forms of the large subunit of the soluble periplasmic [nife] hydrogenase from desulfovibrio gigas have been detected by western analysis. the faster moving form co-migrates with the large subunit from purified, active enzyme. amino acid sequence and composition of the c-terminal tryptic peptide of the large subunit from purified hydrogenase revealed that it is 15 amino acids shorter than that predicted by the nucleotide sequence. processing of the nascent large subunit occurs ...19938405419
flavin dynamics in reduced flavodoxins. a time-resolved polarized fluorescence study.the time-resolved fluorescence and fluorescence anisotropy characteristics of reduced flavin mononucleotide in solution as well as bound in flavodoxins isolated from the bacteria desulfovibrio gigas, desulfovibrio vulgaris, clostridium beijerinckii mp and megasphaera elsdenii have been examined. all fluorescence and fluorescence anisotropy decays were analyzed by two different methods: (a) least-squares fitting with a sum of exponentials and (b) the maximum entropy method to yield distributed li ...19938425547
expression of the gamma-subunit gene of desulfoviridin-type dissimilatory sulfite reductase and of the alpha- and beta-subunit genes is not coordinately regulated.it has been shown [pierik, a. j., duyvis, m. g., van helvoort, j. m. l. m., wolbert, r. b. g. & hagen, w. r. (1992) eur. j. biochem. 205, 111-115] that desulfoviridin, the dissimilatory sulfite reductase of sulfate-reducing bacteria of the genus desulfovibrio, contains a third, gamma, subunit (11 kda), in addition to the well-established alpha (50 kda) and beta (40 kda) subunits, and an alpha 2 beta 2 gamma 2 subunit structure has been proposed. cloning and sequencing of the dsvc gene indicated ...19938436111
cobalamin-mediated mercury methylation by desulfovibrio desulfuricans ls.the prominence of sulfate reducers in mercury biomethylation prompted the examination of the methyl carrier and mercury methylation activity of desulfovibrio desulfuricans ls. there was a low degree of mercury tolerance and a high degree of methylation during fermentative growth; the opposite was true during sulfate reduction. during 2 days of fermentative growth, up to 37% of hgcl2 was methylated at 0.1 micrograms/ml, but only 1.5% was methylated at 10.0 micrograms/ml. less than 1% of the added ...19938439155
complete oxidation of benzoate and 4-hydroxybenzoate by a new sulfate-reducing bacterium resembling desulfoarculus.a new sulfate-reducer "strain sax" was isolated from an anaerobic marine sediment [saxild, denmark]. the isolate was a gram-negative, motile and non-spore-forming rod which sometimes appeared as a curved rod. strain sax differed from all described desulfovibrio-, desulfobotulus- and desulfoarculus-species by the ability to degrade aromatic compounds such as benzoate, 4-hydroxybenzoate and phenol completely to co2. electron donors used included lactate, pyruvate, malate, fumarate, crotonate and b ...19938439232
a novel parameterization scheme for energy equations and its use to calculate the structure of protein molecules.a novel scheme for the parameterization of a type of "potential energy" function for protein molecules is introduced. the function is parameterized based on the known conformations of previously determined protein structures and their sequence similarity to a molecule whose conformation is to be calculated. once parameterized, minima of the potential energy function can be located using a version of simulated annealing which has been previously shown to locate global and near-global minima with ...19938441753
1h and 15n resonance assignments and solution secondary structure of oxidized desulfovibrio vulgaris flavodoxin determined by heteronuclear three-dimensional nmr spectroscopy.sequence-specific 1h and 15n resonance assignments have been made for all 145 non-prolyl residues and for the flavin cofactor in oxidized desulfovibrio vulgaris flavodoxin. assignments were obtained by recording and analyzing 1h-15n heteronuclear three-dimensional nmr experiments on uniformly 15n-enriched protein, ph 6.5, at 300 k. many of the side-chain resonances have also been assigned. observed medium-and long-range noes, in combination with 3jnh alpha coupling constants and 1hn exchange dat ...19938477184
homonuclear and heteronuclear nmr studies of oxidized desulfovibrio vulgaris flavodoxin. sequential assignments and identification of secondary structure elements.recombinant desulfovibrio vulgaris flavodoxin (molecular mass 16.3 kda) was produced in escherichia coli. the oxidized protein has been investigated with a combination of homonuclear and heteronuclear two-dimensional and heteronuclear three-dimensional nmr spectroscopy. sequence-specific assignment of all backbone and most of the side chain 1h and 15n resonances has been obtained. the secondary structure has been inferred from the pattern of sequential, medium-, and long-range noes, together wit ...19938477691
resonance raman studies of iron-only hydrogenases.the nature of the iron-sulfur clusters in oxidized and reduced forms of fe-only hydrogenases from desulfovibrio vulgaris, thermotoga maritima, and clostridium pasteurianum has been investigated by resonance raman spectroscopy. the results indicate the presence of ferredoxin-like [4fe-4s]2+,+ and [2fe-2s]2+,+ clusters in both t. maritima hydrogenase and c. pasteurianum hydrogenase i, but only [4fe-4s]2+,+ clusters in d. vulgaris hydrogenase. this necessitates a reevaluation of the iron-sulfur clu ...19938490025
purification and characterization of an oxygen-labile, nad-dependent alcohol dehydrogenase from desulfovibrio gigas.a nad-dependent, oxygen-labile alcohol dehydrogenase was purified from desulfovibrio gigas. it was decameric, with subunits of m(r) 43,000. the best substrates were ethanol (km, 0.15 mm) and 1-propanol (km, 0.28 mm). n-terminal amino acid sequence analysis showed that the enzyme belongs to the same family of alcohol dehydrogenases as zymomonas mobilis adh2 and bacillus methanolicus mdh.19938491707
nitroaromatic compounds serve as nitrogen source for desulfovibrio sp. (b strain).a sulfate-reducing bacterium, desulfovibrio sp. (b strain), isolated from a continuous anaerobic digester, used various nitroaromatic compounds such as 2,4-dinitrophenol, 2,4-dinitrotoluene, and 2,6-dinitrotoluene as sole nitrogen sources for growth and also used these compounds as electron acceptors in the absence of sulfate in the culture medium. more than 60% of the nitroaromatics were transformed within 6 days of incubation. the organism also used aniline as sole nitrogen source, but not as ...19938500012
analysis of the periplasmic [nife] hydrogenase transcription unit from desulfovibrio fructosovorans.two genes, hyna and hynb, encode the two subunits of the periplasmic [nife] hydrogenase in desulfovibrio fructosovorans. sequencing downstream from hynb revealed a third open reading frame (hync) that has the potential for encoding a polypeptide showing 21% identity with the hyad, hoxm, and hupd proteins, belonging to putative operons encoding escherichia coli hydrogenase 1, alcaligenes eutrophus h16 membrane-bound hydrogenase, and rhizobium leguminosarum uptake hydrogenase, respectively. northe ...19938501043
rubredoxin oxidase, a new flavo-hemo-protein, is the site of oxygen reduction to water by the "strict anaerobe" desulfovibrio gigas.a rubredoxin-oxygen oxidoreductase, a homodimer with a molecular weight of 43 kda per monomer, was found to be a component of an electron transfer chain that couples the reduction of oxygen to water with nadh oxidation. this fad-containing protein appears to contain a new type of heme group. the electron transfer chain is not inhibited by cyanide and azide. in contrast, co decreases nadh oxidation rate and also induces release of the prosthetic groups from the native terminal reductase.19938503894
modeling the structure of pyrococcus furiosus rubredoxin by homology to other x-ray structures.the three-dimensional structure of rubredoxin from the hyperthermophilic archaebacterium, pyrococcus furiosus, has been modeled from the x-ray crystal structures of three homologous proteins from clostridium pasteurianum, desulfovibrio gigas, and desulfovibrio vulgaris. all three homology models are similar. when comparing the positions of all heavy atoms and essential hydrogen atoms to the recently solved crystal structure (day, m. w., et al., 1992, protein sci. 1, 1494-1507) of the same protei ...19938518735
establishing isostructural metal substitution in metalloproteins using 1h nmr, circular dichroism, and fourier transform infrared spectroscopy.far-uv cd, 1h-nmr, and fourier transform infrared (ftir) spectroscopy are three of the most commonly used methods for the determination of protein secondary structure composition. these methods are compared and evaluated as a means of establishing isostructural metal substitution in metalloproteins, using the crystallographically defined rubredoxin from desulfovibrio gigas and its well-characterized cadmium derivative as a model system. it is concluded that analysis of the ftir spectrum of the p ...19958520483
molecular properties of the dissimilatory sulfite reductase from desulfovibrio desulfuricans (essex) and comparison with the enzyme from desulfovibrio vulgaris (hildenborough).the dissimilatory sulfite reductase desulfoviridin was purified from the membrane (msir) and the soluble fraction (ssir) of the sulfate-reducing bacterium desulfovibrio desulfuricans (essex). molecular and spectroscopic properties were determined and compared with the properties of the soluble desulfoviridin from desulfovibrio vulgaris (hildenborough). the enzymes were isolated as alpha 2 beta 2 gamma n (n = 1-3) multimers with a relative molecular mass of 200 +/- 10 (gel filtration). both msir ...19958521853
biodegradation of nitroaromatic compounds.nitroaromatic compounds are released into the biosphere almost exclusively from anthropogenic sources. some compounds are produced by incomplete combustion of fossil fuels; others are used as synthetic intermediates, dyes, pesticides, and explosives. recent research revealed a number of microbial systems capable of transforming or biodegrading nitroaromatic compounds. anaerobic bacteria can reduce the nitro group via nitroso and hydroxylamino intermediates to the corresponding amines. isolates o ...19958561470
purification and characterization of the formate dehydrogenase from desulfovibrio vulgaris hildenborough.formate dehydrogenase from desulfovibrio vulgaris hildenborough, a sulfate-reducing bacterium, has been isolated and characterized. the enzyme is composed of three subunits. a high molecular mass subunit (83,500 da) is proposed to contain a molybdenum cofactor, a 27,000 da subunit is found to be similar to the fe-s subunit of the formate dehydrogenase from escherichia coli and a low molecular mass subunit (14,000 da) holds a c-type heme. the presence of heme c in formate dehydrogenase is reporte ...19958566699
study of the new stability properties induced by amino acid replacement of tyrosine 64 in cytochrome c553 from desulfovibrio vulgaris hildenborough using electrospray ionization mass spectrometry.hydrogen/deuterium exchange as well as charge state distribution monitored by electrospray ionization mass spectrometry were demonstrated to be a powerful and effective new tool for probing conformational properties of proteins in solution. in this paper, the influence of single amino acid replacements on the global conformation of cytochrome c553 from desulfovibrio vulgaris hildenborough using isotopic exchange monitored by electrospray ionization mass spectrometry is reported. based on their r ...19968573183
biochemical studies of the c-type cytochromes of the sulfate reducer desulfovibrio africanus. characterization of two tetraheme cytochromes c3 with different specificity.three c-type cytochromes were isolated and characterized from the sulfate reducer desulfovibrio africanus. a basic tetraheme cytochrome c3 of molecular mass 16 kda was previously described and we have extended its characterization. two other c3-type cytochromes, not previously observed, have also been characterized. these include an acidic tetraheme cytochrome c3 of molecular mass 15 kda and an octaheme dimeric cytochrome c3 with a native size of 35 kda. this is the first report of the presence ...19968573595
sulfate-reducing bacteria in the periodontal pocket.this report is the first to describe the occurrence of sulfate-reducing bacteria in the human mouth. samples of subgingival dental plaque were examined for the presence of sulfate-reducing bacteria. using enrichment cultures, sulfate-reducing bacteria were detected in 25 (58%) of 43 individuals, and in 39 (48%) of the 82 samples. pure isolates of sulfate-reducing bacteria, obtained from a limited number of enrichment cultures, belonged to the genera desulfobacter and desulfovibrio. these genera ...19958596671
kinetics and thermodynamics of the binding of riboflavin, riboflavin 5'-phosphate and riboflavin 3',5'-bisphosphate by apoflavodoxins.the reactions of excess apoflavodoxin from desulfovibrio vulgaris, anabaena variabilis and azotobacter vinelandii with riboflavin 5'-phosphate (fmn), riboflavin 3',5'-bisphosphate and riboflavin are pseudo-first-order. the rates increase with decreasing ph in the range ph 5-8, and, in general, they increase with increasing ionic strength to approach a maximum at an ionic strength greater than 0.4 m. the rate of fmn binding in phosphate at high ph increases to a maximum at an ionic strength of ab ...19968611166
heteronuclear relayed e.cosy applied to the determination of accurate 3j(hn,c') and 3j(h beta,c') coupling constants in desulfovibrio vulgaris flavodoxin.a simple constant-time 3d heteronuclear nmr pulse sequence has been developed to quantitatively determine the heteronuclear three-bond couplings 3j(hn,c') and 3j(h beta,c') in uniformly 13c-enriched proteins. the protocols for measuring accurate coupling constants are based on 1h,13c-heteronuclear relayed e.cosy [schmidt, j.m., ernst, r.r., aimoto, s. and kainosho, m. (1995) j. biomol. nmr, 6, 95-105] in combination with numerical least-squares spectrum evaluation. accurate coupling constants ar ...19968616270
characterization of representative enzymes from a sulfate reducing bacterium implicated in the corrosion of steel.this communication reports the isolation, purification and characterization of key enzymes involved in dissimilatory sulfate reduction of a sulfate reducing bacterium classified as desulfovibrio desulfuricans subspecies desulfuricans new jersey (ncimb 8313) (ddd nj). the chosen strain, originally recovered from a corroding cast iron heat exchanger, was grown in large scale batch cultures. physico-chemical and spectroscopic studies of the purified enzymes were carried out. these analyses revealed ...19968619870
molecular characterization and transcriptional analysis of the putative hydrogenase gene of clostridium acetobutylicum atcc 824.a 2.8-kbp dna region of clostridium acetobutylicum atcc 824 containing the putative hydrogenase gene (hyda) was cloned and sequenced. the 1,745-bp hyda encodes a 64,415-da protein and presents strong identity with the [fe] hydrogenase genes of desulfovibrio and clostridium species. the level of the putative hyda mrna was high in cells from an acidogenic or an alcohologenic phosphate-limited continuous culture, while it was comparatively very low in cells from a solventogenic phosphate-limited co ...19968626337
the structure of desulfovibrio vulgaris rubrerythrin reveals a unique combination of rubredoxin-like fes4 and ferritin-like diiron domains.we have determined the structure of rubrerythrin, a non-haem iron protein from the anaerobic sulphate-reducing bacterium, desulfovibrio vulgaris (hildenborough), by x-ray crystallography. the structure reveals a tetramer of two-domain subunits. each subunit contains a four-helix bundle surrounding a diiron-oxo site and a c-terminal rubredoxin-like fes4 domain. the diiron-oxo site contains a larger number of carboxylate ligands and a higher degree of solvent exposure than do those in other diiron ...19968646540
similarities in the architecture of the active sites of ni-hydrogenases and fe-hydrogenases detected by means of infrared spectroscopy.three groups that absorb in the 2100-1800-cm-1 infrared spectral region have recently been detected in ni-hydrogenase from chromatium vinosum [bagley, k.a., duin, e.c., roseboom, w., albracht, s. p.j. & woodruff, w.h. (1995) biochemistry 34, 5527-5535]. to assess the significance and generality of this observation, we have carried out an infrared-spectroscopic study of eight hydrogenases of three different types (nickel, iron and metal-free) and of 11 other iron-sulfur and/or nickel proteins. in ...19968647106
an nmr-derived model for the solution structure of oxidized thermotoga maritima 1[fe4-s4] ferredoxin.the solution structure of the 60-residue 1[fe4-s4] ferredoxin from the hyperthermophilic bacterium thermotoga maritima was determined based on 683 distance and 35 dihedral angle restraints that were obtained from nmr data. in addition, data known from crystallographic studies of ferredoxins was used for modeling of the iron-sulfur cluster and its environment. the protein shows a globular fold very similar to the fold of the related 1[fe4-s4] ferredoxins from desulfovibrio gigas and desulfovibrio ...19968647119
primary structure of desulfoferrodoxin from desulfovibrio desulfuricans atcc 27774, a new class of non-heme iron proteins.the primary structure of desulfoferrodoxin from desulfovibrio desulfuricans atcc 27774, a redox protein with two mononuclear iron sites, was determined by automatic edman degradation and mass spectrometry of the composing peptides. it contains 125 amino acid residues of which five are cysteines. the first four, cys-9, cys-12, cys-28 and cys-29, are responsible for the binding of center i which has a distorted tetrahedral sulfur coordination similar to that found in desulforedoxin from d. gigas. ...19968647238
regulation of the redox order of four hemes by ph in cytochrome c3 from d. vulgaris miyazaki f.the assignment of 1h-nmr signals of the heme methyl and propionate groups of cytochrome c3 of d. vulgaris miyazaki f was performed. the heme assignment was revised for hemes 2 and 3 (sequential heme numbering). namely, heme 4 is mainly reduced at first with hemes 1, 2 and 3 following it in this order. the p2h titration of heme methyl signals in four macroscopic oxidation states was performed in the p2h range of 5.2 to 9.0. while the heme methyl resonances in the fully oxidized state showed just ...19968652627
expression in escherichia coli and characterization of a reconstituted recombinant 7fe ferredoxin from desulfovibrio africanus.desulfovibrio africanus ferredoxin iii is a monomeric protein (molecular mass of 6585 da) that contains one [3fe-4s]1+/0 and one [4fe-4s]2+/1+ cluster when isolated aerobically. the amino acid sequence consists of 61 amino acids, including seven cysteine residues that are all involved in co-ordination to the clusters. in order to isolate larger quantities of d. africanus ferredoxin iii, we have overexpressed it in escherichia coli by constructing a synthetic gene based on the amino acid sequence ...19968660311
purification and characterization of the hydrogenase from thiobacillus ferrooxidans.hydrogenase of thiobacillus ferrooxidans atcc 19859 was purified from cells grown lithoautotrophically with 80% hydrogen, 8.6% carbon dioxide, and 11.4% air. hydrogenase was located in the 140,000 x g supernatant in cell-free extracts. the enzyme was purified 7.3-fold after chromatography on procion red and q-sepharose with a yield of 19%, resulting in a 85% pure preparation with a specific activity of 6.0 u (mg protein)-1. with native page, a mol. mass of 100 and 200 kda was determined. with sd ...19968661919
total synthesis of a miniferredoxin.a miniferredoxin has been designed based on the desulfovibrio gigas ferredoxin ii structure and has been successfully synthesized. the 31 amino acid apoprotein was synthesized via standard fmoc solid phase peptide synthesis and in vitro cluster insertion carried out. the uv-visible spectrum of the miniferredoxin (peak at 300 nm and a shoulder at 405 nm) shows the same features as that of the d. gigas ferredoxin. cyclic voltammetry indicated a quasireversible electrode process with a midpoint pot ...19968670287
tungsten in biological systems.tungsten (atomic number 74) and the chemically analogous and very similar metal molybdenum (atomic number 42) are minor yet equally abundant elements on this planet. the essential role of molybdenum in biology has been known for decades and molybdoenzymes are ubiquitous. yet, it is only recently that a biological role for tungsten has been established in prokaryotes, although not as yet in eukaryotes. the best characterized organisms with regard to their metabolism of tungsten are certain specie ...19968672295
nmr studies of flavoproteins. 19968674598
regulation of the redox potentials of flavodoxins: modification of the flavin binding. 19968674606
the effects of ph on the absorption spectrum of the hydroquinone of flavodoxin from desulfovibrio vulgaris. 19968674675
the redox potentials of flavodoxin from desulfovibrio vulgaris and ferredoxin-nadp+ reductase from spinacia oleracea and their complexes. 19968674686
reaction of apoflavodoxin from desulfovibrio vulgaris with 5,5'-dithio-bis-(2-nitrobenzoic acid). 19968674695
nmr investigation of the solution conformation of oxidized flavodoxin from desulfovibrio vulgaris. determination of the tertiary structure and detection of protein-bound water molecules.desulfovibrio vulgaris flavodoxin has been investigated with a combination of homo- and hetero-nuclear two-dimensional and three-dimensional nmr spectroscopy. the analysis of noe, hydrogen exchange and j-coupling data led to a set of 1349 noe, 63 hydrogen bond and 109 backbone phi-angle restraints which were used to determine the solution structure of the oxidized flavodoxin applying the distance geometry program diana combined with restrained energy minimization methods. flavodoxin in solution ...19968681954
sulfonates: novel electron acceptors in anaerobic respiration.the enrichment and isolation in pure culture of a bacterium, identified as a strain of desulfovibrio, able to release and reduce the sulfur of isethionate (2-hydroxyethanesulfonate) and other sulfonates to support anaerobic respiratory growth, is described. the sulfonate moiety was the source of sulfur that served as the terminal electron acceptor, while the carbon skeleton of isethionate functioned as an accessory electron donor for the reduction of sulfite. cysteate (alanine-3-sulfonate) and s ...19968703197
dissimilatory sulfite reductase revisited. the desulfoviridin molecule does contain 20 iron ions, extensively demetallated sirohaem, and an s = 9/2 iron-sulfur cluster.assimilatory sulfite reductase contains a sirohaem that is very weakly coupled to a [4fe-4s] cubane, i.e. five iron atoms in total. dissimilatory sulfite reductase is a complex system with 20 fe atoms/alpha 2 beta 2 gamma 2 hexamer. a recent revision of the purification procedure for the desulfovibrio vulgaris dissimilatory enzyme has afforded a preparation of only 10 fe atoms hexamer, this has led to the convulsion that the topology of prosthetic groups parallels that of the assimilatory system ...19968706673
crystal structure of flavodoxin from desulfovibrio desulfuricans atcc 27774 in two oxidation states.the crystal structures of the flavodoxin from desulfovibrio desulfuricans atcc 27774 have been determined and refined for both oxidized and semi-reduced forms to final crystallographic r-factors of 17.9% (0.8-0.205-nm resolution) and 19.4% (0.8-0.215-nm resolution) respectively. native flavodoxin crystals were grown from ammonium sulfate with cell constants a = b = 9.59 nm, c=3.37nm (oxidized crystals) and they belong to space group p3(2)21. semireduced crystals showed some changes in cell dimen ...19968706707
structural and functional characterization of cytochrome c3 from d. desulfuricans atcc 27774 by 1h-nmr.cooperativity between redox and protonation centres is known to be crucial for the function of complex proteins, but it is often difficult to describe in terms of thermodynamic parameters. cytochrome c3 is a good model for these studies since, while retaining the overall complexity of larger systems, it is suitable for detailed crystallographic and spectroscopic studies. assignment of the haem substituent nmr resonances, together with nmr redox titrations of cytochrome c3 from d. desulfuricans a ...19968706829
nitrite reductase from desulfovibrio desulfuricans (atcc 27774)--a heterooligomer heme protein with sulfite reductase activity.the membrane bound cytochrome c nitrite reductase from the sulfate reducer desulfovibrio desulfuricans (atcc 27774) was found to have a high specific activity in the reduction of sulfite, producing stoichiometric amounts of sulfide. the k(m) for sulfite in the mv+.:sulfite oxidoreductase assay is 0.75 mm, and the specific activity 2.06 mumolh2/min/mg. visible and epr spectroscopies studies indicate that the enzyme high-spin heme reacts with sulfite in the oxidised state, and that sulfide partial ...19968713097
1h nmr investigation of the secondary structure, tertiary contacts and cluster environment of the four-iron ferredoxin from the hyperthermophilic archaeon thermococcus litoralis.the solution molecular structure of the four-iron ferredoxin (fd) from the hyperthermophilic archaeon thermococcus litoralis (tl) has been investigated by 1h nmr spectroscopy. tocsy and noesy experiments in h2o, tailored to detect both weakly and strongly relaxed resonances, together with steady-state noes in both h2o and d2o, allowed the identification of 58 of the 59 residues, with one residue near the paramagnetic center undetected. it is shown that the contact shifted and strongly relaxed si ...19968720830
crystal structure of a dimeric octaheme cytochrome c3 (m(r) 26,000) from desulfovibrio desulfuricans norway.the octaheme cytochrome c3 (m(r) 26,000; cc3) from desulfovibrio desulfuricans norway is a dimeric cytochrome made up of two identical subunits, each containing four heme groups. it is involved in the redox transfer chain of sulfate-reducing bacteria, which links the periplasmic oxidation of hydrogen to the cytoplasmic reduction of sulfate. the amino-acid sequence of cc3 shows similarities to that of the tetraheme cytochrome c3 (m(r) 13,000; c3) from the same bacteria. structural analysis of cc3 ...19968740362
[isolation and reassociation of acetogen and methanogen in a syntrophobic coculture degrading butyrate anaerobically].anaerobic coculture bf2 which degraded butyrate into acetate and produced methane was isolated from granular methanogenic sludge. the coculture is associated syntrophically the syntrophomonas subsp. saponavida strain cf2 with methanobacterium formicicum strain mf2 and appeared to degraded c4 approximately c18 fatty acids including isobutyrate. the optimal temperature and ph for growth was 37 degrees c and 7.7 respectively. the strain cf2 was obtained in pure culture with crotonate as substrate a ...19958745550
genetic control of the resistance to phage c1 of escherichia coli k-12.escherichia coli k-12 lytic phage c1 was earlier isolated in our laboratory. its adsorption is controlled by at least three bacterial genes: dcra, dcrb, and btub. our results provide evidence that the dcra gene located at 60 min on the e. coli genetic map is identical to the sdac gene. this gene product is an inner membrane protein recently identified as a putative specific serine transporter. the dcrb gene, located at 76.5 min, encodes a 20-kda processed periplasmic protein, as determined by ma ...19968752353
preliminary crystallographic analysis of the oxidized form of a two mono-nuclear iron centres protein from desulfovibrio desulfuricans atcc 27774.crystals of the fully oxidized form of desulfoferrodoxin were obtained by vapor diffusion from a solution containing 20% peg 4000, 0.1 m hepes buffer, ph 7.5, and 0.2 m cacl2. trigonal and/or rectangular prisms could be obtained, depending on the temperature used for the crystal growth. trigonal prisms belong to the rhombohedral space group r32, with a = 112.5 a and c = 63.2 a; rectangular prisms belong to the monoclinic space group c2, with a = 77.7 a, b = 80.9 a, c = 53.9 a, and beta = 98.1 de ...19968762151
investigation of oxidation state-dependent conformational changes in desulfovibrio vulgaris hildenborough cytochrome c553 by two-dimensional h-nmr spectra.two-dimensional nuclear magnetic resonance spectroscopy (2d-nmr) was used to assign the proton resonances of ferricytochrome c553 from desulfovibrio vulgaris hildenborough. the spin systems of 76 out of 79 amino acids were identified by j-correlation spectroscopy (cosy and hohaha) in h20 and d20 and correlated by nuclear overhauser effect spectroscopy (noesy). the proton chemical shifts are compared in both oxidized and reduced states of the protein at 23 degrees c and ph 5.9. chemical shift var ...19968766830
properties of xanthine dehydrogenase variants from rosy mutant strains of drosophila melanogaster and their relevance to the enzyme's structure and mechanism.xanthine dehydrogenase, a molybdenum, iron-sulfur flavoenzyme encoded in the fruit fly drosophila melanogaster by the rosy gene, has been characterised both from the wild-type and mutant files. enzyme assays, using a variety of different oxidising and reducing substrates were supplemented by limited molecular characterisation. four rosy strains showed no detectable activity in any enzyme assay tried, whereas from four wild-type and three rosy mutant strains, those for the [e89k], [l127f] and [l1 ...19968774727
desulfovibrio gabonensis sp. nov., a new moderately halophilic sulfate-reducing bacterium isolated from an oil pipeline.two moderately halophilic sulfate-reducing bacteria were isolated from an african oil pipeline and designated strains sebr 3640 and sebr 2840t (t = type strain). both of these strains possess traits that define the genus desulfovibrio. the cells of both isolates were motile curved rods that had a single polar flagellum and contained desulfoviridin, and both isolates utilized lactate, pyruvate, malate, fumarate, succinate, and ethanol in the presence of sulfate. sulfite, thiosulfate, and elementa ...19968782680
probable new species of desulfovibrio isolated from a pyogenic liver abscess.a fastidious, slowly growing, spiral gram-negative bacterium was isolated from the liver abscess of an 82-year-old man with a 3-week history of febrile illness. the organism was an obligate anaerobe that grew at 37 and 42 degrees c but not at 25 degrees c. its vibrioid or spiral morphology on gram staining, rapid progressive motility, electron micrograph features, and biochemical tests were all consistent with the organism belonging to the genus desulfovibrio. 16s rrna gene sequencing of this or ...19968784584
1h and 15n nmr resonance assignments and solution secondary structure of oxidized desulfovibrio desulfuricans flavodoxin.sequence-specific 1h and 15n resonance assignments have been made for 137 of the 146 nonprolyl residues in oxidized desulfovibrio desulfuricans [essex 6] flavodoxin. assignments were obtained by a concerted analysis of the heteronuclear three-dimensional 1h-15n noesy-hmqc and tocsy-hmqc data sets, recorded on uniformly 15n-enriched protein at 300 k. numerous side-chain resonances have been partially or fully assigned. residues with overlapping 1hn chemical shifts were resolved by a three-dimensi ...19968785498
population dynamics of propionate-oxidizing bacteria under methanogenic and sulfidogenic conditions in anaerobic granular sludge.laboratory-scale upflow anaerobic sludge-bed reactors were inoculated with industrial granular sludge and fed with either propionate or propionate and sulfate. the population dynamics of the propionate-oxidizing bacteria desulfobulbus sp. and the syntrophically growing strain syn7 were studied in reactors by dot blot and in situ hybridization with 16s rrna-based oligonucleotide probes.19968787413
intraspecies variability of cellular fatty acids among soil and intestinal strains of desulfovibrio desulfuricans.a comparison of cellular fatty acid profiles of desulfovibrio desulfuricans dsm 642 and 14 wild strains of this species, isolated from two completely different environments, soil and the human intestine, was carried out. all the d. desulfuricans strains grown on lactate and sulfate indicated the presence of considerable amounts of i-c15:0, i-c17:1 and c16:0. although differences in the quantities of individual fatty acids present in each strain were clear in the group of soil strains (similarity ...19968795227
cloning, expression, and sequence analysis of the three genes encoding quinoline 2-oxidoreductase, a molybdenum-containing hydroxylase from pseudomonas putida 86.the three genes coding for quinoline 2-oxidoreductase (qor) of pseudomonas putida 86 were cloned and sequenced. the qor genes are clustered in the transcriptional order medium (m) small (s), large (l) and code for three subunits of 288 (qorm), 168 (qors), and 788 (qorl) amino acids, respectively. formation of active quinoline 2-oxidoreductase and degradation of quinoline occurred in a recombinant p. putida kt2440 clone. the amino acid sequences of qor show significant homology to various prokary ...19968798497
redox properties of cytochrome c nitrite reductase from desulfovibrio desulfuricans atcc 27774.the dissimilatory nitrite reductase from desulfovibrio desulfuricans atcc 27774 catalyzes the reduction of nitrite to ammonia. previous spectroscopic investigation revealed that it is a hexaheme cytochrome containing one high spin ferric heme and five low spin ferric hemes in the oxidized enzyme. the current study uses the high resolution of mössbauer spectroscopy to obtain redox properties of the six heme groups. correlating the mössbauer findings with the epr data reveals the pairwise spin-spi ...19968798514
interactions of 77se and 13co with nickel in the active site of active f420-nonreducing hydrogenase from methanococcus voltae.the selenium-containing f420-nonreducing hydrogenase from methanococcus voltae was prepared in the nia(i) middle dotco state. the effect of illumination on this light-sensitive species was studied. epr studies were carried out with enzyme containing natural selenium or with enzyme enriched in 77se. samples were prepared with either co or 13co. in the nia(i) middle dotco state, the nuclear spins of both 77se (i = 1/2) and 13c (i = 1/2) interacted with the nickel-based unpaired electron, suggestin ...19968798608
a structure-based catalytic mechanism for the xanthine oxidase family of molybdenum enzymes.the crystal structure of the xanthine oxidase-related molybdenum-iron protein aldehyde oxido-reductase from the sulfate reducing anaerobic gram-negative bacterium desulfovibrio gigas (mop) was analyzed in its desulfo-, sulfo-, oxidized, reduced, and alcohol-bound forms at 1.8-a resolution. in the sulfo-form the molybdenum molybdopterin cytosine dinucleotide cofactor has a dithiolene-bound fac-[mo, = o, = s, ---(oh2)] substructure. bound inhibitory isopropanol in the inner compartment of the subs ...19968799115
cytochrome c3 from desulfovibrio gigas: crystal structure at 1.8 a resolution and evidence for a specific calcium-binding site.crystals of the tetraheme cytochrome c3 from sulfate-reducing bacteria desulfovibrio gigas (dg) (mw 13 kda, 111 residues, four heme groups) were obtained and x-ray diffraction data collected to 1.8 a resolution. the structure was solved by the method of molecular replacement and the resulting model refined to a conventional r-factor of 14.9%. the three-dimensional structure shows many similarities to other known crystal structures of tetraheme c3 cytochromes, but it also shows some remarkable di ...19968819167
comparison of low oxidoreduction potential cytochrome c553 from desulfovibrio vulgaris with the class i cytochrome c family.the cytochrome c553 from desulfovibrio vulgaris (dvh c553) is of importance in the understanding of the relationship of structure and function of cytochrome c due to its lack of sequence homology with other cytochromes, and its abnormally low oxido-reduction potential. in evolutionary terms, this protein also represents an important reference point for the understanding of both bacterial and mitochondrial cytochromes c. using the recently determined nuclear magnetic resonance (nmr) structure of ...19968820485
characterization of microbial communities in anaerobic bioreactors using molecular probes.the microbial community structure of twenty-one single-phase and one two-phase full-scale anaerobic sewage sludge digesters was evaluated using oligonucleotide probes complementary to conserved tracts of the 16s rrnas of phylogenetically defined groups of methanogens and sulfate-reducing bacteria. these probe results were interpreted in combination with results from traditional chemical analyses and metabolic activity assays. it was determined that methanogens in "healthy" mesophilic, single-pha ...19958821785
evaluation of the electrostatic effect of the 5'-phosphate of the flavin mononucleotide cofactor on the oxidation--reduction potentials of the flavodoxin from desulfovibrio vulgaris (hildenborough).two mutants of the desulfovibrio vulgaris flavodoxin, t12h and n14h, were generated which, for the first time, place a basic residue within the normally neutral 5'-phosphate binding loop of the flavin mononucleotide cofactor binding site found in all flavodoxins. these histidine residues were designed to form an ion pair with the dianionic 5'-phosphate, either altering its ionization state or offsetting its negative charge to allow evaluation of the magnitude of its electrostatic effect on the r ...19968823179
the dcr gene family of desulfovibrio: implications from the sequence of dcrh and phylogenetic comparison with other mcp genes.desulfovibrio vulgaris hildenborough contains a family of genes for methyl-accepting chemotaxis proteins (mcps). here we report the complete sequence of the gene for desulfovibrio chemoreceptor h (dcrh). the deduced amino acid sequence of dcrh protein, which has an enlarged n-terminal, ligand binding domain, indicates a structure similar to that of other mcps. comparison of the sequences for dcra, determined earlier, and dcrh indicated that similarity is essentially limited to the c-terminal exc ...19968836438
degradative capacities and 16s rrna-targeted whole-cell hybridization of sulfate-reducing bacteria in an anaerobic enrichment culture utilizing alkylbenzenes from crude oil.a mesophilic sulfate-reducing enrichment culture growing anaerobically on crude oil was used as a model system to study which nutritional types of sulfate-reducing bacteria may develop on original petroleum constituents in oil wells, tanks, and pipelines. chemical analysis of oil hydrocarbons during growth revealed depletion of toluene and o-xylene within 1 month and of m-xylene, o-ethyltoluene, m-ethyltoluene, m-propyltoluene, and m-isopropyltoluene within approximately 2 months. in anaerobic c ...19968837415
transposon mutagenesis in desulfovibrio desulfuricans: development of a random mutagenesis tool from tn7.the transposons tn5, tn7, tn9, and tn10 or their derivatives have been examined for transposition in the sulfate-reducing bacterium desulfovibrio desulfuricans g20. tn7 inserted with a frequency of 10(-4) to 10(-3) into a unique attachment site that shows strong homology with those sites identified in other gram-negative bacteria. inactivation of the tnsd gene in tn7, encoding the function directing insertion into the unique site, yielded a derivative that transposed essentially randomly with a ...19968837431
new conformational properties induced by the replacement of tyr-64 in desulfovibrio vulgaris hildenborough ferricytochrome c553 using isotopic exchanges monitored by mass spectrometry.in order to study the conformational stability induced by the replacement of tyr-64 in desulfovibrio vulgaris hildenborough (dvh) cytochrome c553, fast peptic digestion of deuterated protein followed by separation and measurement of related peptides using liquid chromatography coupled to electrospray ionization mass spectrometry was performed. we show that the h-bonding and/or solvent accessibility properties were modified by the single-site mutation. the mutant proteins can be classified into t ...19968849688
activation and degradation of benzoate, 3-phenylpropionate and crotonate by syntrophus buswellii strain ga. evidence for electron-transport phosphorylation during crotonate respiration.a strictly anaerobic, benzoate-degrading bacterium, syntrophus buswellii strain ga, was able to degrade benzoate or 3-phenylpropionate to acetate, co2 and h2, if the hydrogen partial pressure was sufficiently low. the hydrogen was removed in syntrophic coculture by methanospirillum hungatei or by desulfovibrio sp. through interspecies hydrogen transfer or in pure culture by the use of crotonate as reducible cosubstrate. alternatively, s. buswellii strain ga could grow in pure culture with croton ...19968867639
characterization of sulfate-reducing bacteria isolated from oil-field waters.the occurrence and metabolic capacities of sulfate-reducing bacteria (srb) were studied in 23 water samples taken from producing wells at 14 different sites. oil fields in france, the north sea, and the gulf of guinea were selected and classified according to physicochemical parameters (salinity ranging from 0.3 to 120 g.l-1 and temperature between 29 and 85 degrees c). after the distribution of srb within oil fields was studied, several strains of srb were isolated and characterized metabolical ...19968868233
electron transfer in tetrahemic cytochromes c3: spectroelectrochemical evidence for a conformational change triggered by heme iv reduction.electron transfer in tetrahemic cytochromes c3 from desulfovibrio vulgaris hildenborough (d.v.h.) and desulfovibrio desulfuricans norway (d.d.n.) strains has been investigated by thin layer spectroelectrochemistry with visible absorption, cd, and resonance raman (rr) monitoring. the observed splitting of the isosbestic point in the soret absorption band indicates that the electron transfer from the (feiii)4 state to the (feii)4 state proceeds via an intermediate species, which corresponds to 25 ...19968873609
rnase e can inhibit the decay of some degradation intermediates: degradation of desulfovibrio vulgaris cytochrome c3 mrna in e coli.in escherichia coli, ribonuclease e (rnase e) is a key endonuclease in mrna decay. we have analysed the role of e coli rnase e on the degradation of a heterologous cytochrome c3 (cyc) mrna from desulfovibrio vulgaris hildenborough. the decay of the cyc transcript in wild-type and mutant e coli cells was followed and the degradation intermediates analysed by northern blotting and s1 protection analysis. the half-life of total cyc mrna intermediates was increased in the rnase e mutant. a number of ...19968874797
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