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genetics of hydrogenase from aerobic lithoautotrophic bacteria.aerobic facultatively autotrophic hydrogen bacteria are distinguished on the basis of their hydrogen-oxidizing enzyme system (hox). the major group, represented by paracoccus denitrificans and pseudomonas facilis, contains a membrane-bound, electron transport-coupled protein. species of nocardia are characterized by the possession of a cytoplasmic nad-dependent hydrogenase. both enzymes are present in strains of alcaligenes. all hydrogenases from lithoautotrophs are h2-consuming nickel-iron-sulf ...19863089306
cytochrome c oxidase from paracoccus denitrificans in triton x-100: aggregation state and kinetics.cytochrome c oxidase from paracoccus denitrificans was homogeneously dispersed in triton x-100. using gel exclusion chromatography and sucrose gradient centrifugation analysis a molecular weight of the detergent-protein complex of 155,000 was determined. after subtraction of the bound detergent (111 mol/mol heme aa3) a molecular weight of 85,000 resulted, which agreed well with the model of a monomer containing two subunits. this monomer showed high cytochrome c oxidase activity when measured sp ...19863017928
immunological identification of aa3-type cytochrome oxidase in membrane preparations of the cyanobacterium anacystis nidulans.membranes were isolated from the cyanobacterium anacystis nidulans by french press extrusion of lysozyme-treated cells. the membranes were solubilized with sodium dodecylsulfate and subjected to denaturing polyacrylamide gel electrophoresis. separated polypeptides were transferred to nitrocellulose by western blotting, and incubated with antibodies against aa3-type cytochrome oxidase of paracoccus denitrificans; antibodies against subunits i and ii, and against the holoenzyme, were used and gave ...19863010964
cytochrome c oxidase from paracoccus denitrificans. 19862856122
isolation of ubiquinol oxidase from paracoccus denitrificans. 19862856133
isolation of a three-subunit cytochrome bc1 complex from paracoccus denitrificans. 19862856134
a simple, one-step purification for cytochrome b from the bc1 complexes of bacteria. 19862856136
characterisation of phosphate binding to mitochondrial and bacterial membrane-bound atp synthase by studies of inhibition with 4-chloro-7-nitrobenzofurazan.the effect of phosphate on the inhibition by 4-chloro-7-nitrobenzofurazan of the atpase activity of the proton-translocating atp synthase in heart submitochondrial particles was investigated. binding of phosphate protected strongly against the inhibition. a dissociation constant of 0.2 mm was determined for the enzyme x pi complex and shown to be independent of ph in the range 7.0-8.0. the protective effect of phosphate was mimicked by arsenate but not by sulphate or malonate. similar results we ...19862869972
electron-transfer reactions between flavodoxin semiquinone and c-type cytochromes: comparisons between various flavodoxins.as an extension of previous work from this laboratory using clostridium pasteurianum flavodoxin [tollin, g., cheddar, g., watkins, j. a., meyer, t. e., & cusanovich, m. a. (1984) biochemistry 23, 6345-6349], we have measured the rate constants as a function of ionic strength for electron transfer from the semiquinones of clostridium mp, anacystis nidulans, and azotobacter vinelandii flavodoxins to the following oxidants: cytochrome c from tuna and horse, paracoccus denitrificans cytochrome c2, p ...19863024711
characterization of two inducible periplasmic c-type cytochromes from paracoccus denitrificans.when grown on methanol or methylamine, paracoccus denitrificans synthesized three periplasmic soluble c-type cytochromes, cytochrome c550 and two additional cytochromes which were not present during growth on succinate and have not previously been characterized. these cytochromes have been separated from each other and their physical properties have been determined. the inducible cytochromes, c551i and c553i, exhibit mr and pi values of 22,000 and 3.5, and 30,000 and 3.8, respectively. cytochrom ...19863013855
evolutionary relationship of denitrifying bacteria as deduced from 5s rrna sequences.the nucleotide sequences of 5s rrna from seven denitrifying bacteria have been determined. based on these sequences and those reported in the literature (including two denitrifiers), a phylogenic tree of 104 eubacterial 5s rrna sequences has been constructed to establish the position of the denitrifying bacteria. these bacteria belong to either one of the three major subgroups of gram-negative bacteria. the grouping based on 5s rrna sequences is almost compatible with the type of the nitrite red ...19862434470
properties of paracoccus denitrificans amicyanin.paracoccus denitrificans synthesizes an inducible, periplasmic, blue copper protein [husain, m., & davidson, v.l. (1985) j. biol. chem. 260, 14626-14629] that can be classified as an amicyanin on the basis of its ability to accept electrons from methylamine dehydrogenase. the amino acid composition and sequence of the 10 n-terminal residues of this protein have been determined. from these data, it is evident that amicyanin is structurally distinct from azurins as it contains no disulfide bond an ...19863718960
the respiratory nitrate reductase from paracoccus denitrificans. molecular characterisation and kinetic properties.the respiratory nitrate reductase from paracoccus denitrificans has been purified in the non-ionic detergent nonidet p-40. the enzyme comprises three polypeptides, alpha, beta and gamma with estimated relative molecular masses of 127 000, 61 000 and 21 000. duroquinol or reduced-viologen compounds acted as the reducing substrates. the nitrate reductase contained a b-type cytochrome that was reduced by duroquinol and oxidised by nitrate. a preparation of the enzyme that lacked both detectable b-t ...19863732277
electron transfer flavoprotein from methylophilus methylotrophus: properties, comparison with other electron transfer flavoproteins, and regulation of expression by carbon source.when grown on methylated amines as a carbon source, methylophilus methylotrophus synthesizes an electron transfer flavoprotein (etf) which is the natural electron acceptor of trimethylamine dehydrogenase. it is composed of two dissimilar subunits of 38,000 and 42,000 daltons and 1 mol of flavin adenine dinucleotide. it was reduced by trimethylamine dehydrogenase to a stable anionic semiquinone form, which could not be converted, either enzymatically or chemically, to the fully reduced dihydroqui ...19863711024
iron-dependent production of a heat-modifiable, 23,000-mr outer membrane protein in paracoccus denitrificans.production of a 23,000-mr major outer membrane protein of paracoccus denitrificans atcc 13543 was dependent upon the addition of iron to a succinate-salts medium. the 23,000-mr protein was not produced in an iron-deficient medium, but production of five outer membrane proteins in the 85,000- to 72,000-mr range and of catechol were induced. the 23,000-mr protein was not produced in a complex medium even when ferric citrate was added to the medium. production of the protein was influenced by the c ...19863759911
structural relationships between the nadh dehydrogenases of paracoccus denitrificans and bovine heart mitochondria as revealed by immunological cross-reactivities.an antibody raised against two subunits (mr 48 000 and 25 000) of nadh dehydrogenase from paracoccus denitrificans cross-reacts with one of more than 20 polypeptides that form the bovine heart mitochondrial nadh dehydrogenase. the cross-reacting subunit has mr 51 000 and is believed to be the nadh-binding subunit of the enzyme. antibodies raised against certain subunits of the bovine heart nadh dehydrogenase were tested for cross-reactivity with p. denitrificans cytoplasmic membranes. of those t ...19863956724
modeling of microbial substrate conversion, growth and product formation in a recycling fermentor.paracoccus denitrificans and bacillus licheniformis were grown in a carbon- and energy source-limited recycling fermentor with 100% biomass feedback. experimental data for biomass accumulation and product formation as well as rates of carbon dioxide evolution and oxygen consumption were used in a parameter optimization procedure. this procedure was applied on a model which describes biomass growth as a linear function of the substrate consumption rate and the rate of product formation as a linea ...19863532947
a novel prenyltransferase from paracoccus denitrificans.a new polyprenyltransferase catalysing the formation of z-double bonds was found and partially purified from extracts of paracoccus denitrificans. the enzyme catalysed a consecutive condensation of isopentenyl diphosphate with ee-farnesyl diphosphate as a primer to produce ee-farnesyl-all-z-hexaprenyl diphosphate (ze-mixed nonaprenyl diphosphate) as the final product. not only ee-farnesyl diphosphate but also neryl diphosphate, ze-farnesyl diphosphate, zee-geranylgeranyl diphosphate and zzee-pen ...19863707524
succinate dehydrogenase in rhodopseudomonas sphaeroides: subunit composition and immunocross-reactivity with other related bacteria.antibodies were raised against the succinate dehydrogenase (sdh) present in the chromatophores of phototrophically grown rhodopseudomonas sphaeroides. crossed immunoelectrophoresis experiments indicated that the sdh present in the cytoplasmic membranes of heterotrophically grown r. sphaeroides is probably the same enzyme observed in the chromatophores. the enzyme was extracted by triton x-100 in a form which consisted of only two subunits (molecular weight, 68,000 and 30,000) and was not associa ...19853874866
statistical analysis of growth limitations in paracoccus denitrificans: an experiment with a completely randomized two-way factorial design with replications.cultivation of microorganisms under growth limitation is a widely used technique in microbiology. the relevant investigations, though seemingly based on straightforward experiments, have generated conflicting results, e.g., concerning molar growth yields. the purpose of this paper is to show that discrepancies in the literature could be solved if more attention were paid to methodology, especially statistics. new experimental results, concerning growth limitations in paracoccus denitrificans, wi ...19854004200
on the dielectrically observable consequences of the diffusional motions of lipids and proteins in membranes. 2. experiments with microbial cells, protoplasts and membrane vesicles.the dielectric properties of suspensions of intact cells of methylophilus methylotrophus, paracoccus denitrificans and bacillus subtilis have been measured in the frequency range 1 khz to 13 mhz. all possess a pronounced dispersion corresponding in magnitude and relaxation time to the "beta-dispersion" in a terminology defined by schwan [adv. biol. med. phys. 5:147-209 (1957)]. the latter two strains, but not m. methylotrophus, also possess a substantial alpha-dispersion. the relaxation time of ...19853935420
the primary structures of pseudomonas am1 amicyanin and pseudoazurin. two new sequence classes of blue copper proteins.the amino acid sequences of two blue copper proteins from the pink facultative methylotroph pseudomonas am1 (n.c.i.b. 9133) were determined. they each consist of a single polypeptide chain and bind one copper atom. amicyanin contains 99 and pseudoazurin 123 residues. copper-binding sites, consisting of the side chains of two histidine, one cysteine and one methionine residues, can be recognized in each protein by analogy with azurin and plastocyanin, but the spacings of the ligand residues are d ...19854091802
isolation and characterization of paracoccus denitrificans mutants with defects in the metabolism of one-carbon compounds.mutants deficient in the metabolism of one-carbon compounds have been obtained by treating paracoccus denitrificans with the mutagen n-methyl-n'-nitro-n-nitrosoguanidine. mutants were selected without enrichment procedures by newly developed plate screening tests. the obtained mutants were characterized by their growth responses, cytochrome composition, enzyme activities, and immunogenic reaction with antisera against methanol dehydrogenase. by these criteria five mutant classes could be disting ...19853905763
on the structure of heme d1. an isobacteriochlorin derivative as the prosthetic group of dissimilatory nitrite reductase?timkovich and co-workers have recently proposed a chlorin macrocycle structure for the heme d1 prosthetic group isolated from cytochrome cd1 of pseudomonas aeruginosa and paracoccus denitrificans (timkovich, r., cork, m. s., and taylor, p. v. (1984) j. biol. chem. 259, 1577-1585; 15089-15093). however, this chlorin structure deduced by them is not entirely consistent with the spectral data. an alternative structure is proposed here based on the available spectral evidence. it is suggested that h ...19853926768
nitric oxide-dependent proton translocation in various denitrifiers.respiration of no resulted in transient proton translocation in anaerobically grown cells of four physiologically diverse denitrifiers. paracoccus denitrificans, rhodopseudomonas sphaeroides subsp. denitrificans, "achromobacter cycloclastes," and rhizobium japonicum gave, respectively, h+/no ratios of 3.65, 4.96, 1.94, and 1.12. antimycin a completely inhibited no-dependent proton translocation in p. denitrificans and severely restricted translocation in the r. sphaeroides strain. proton uptake ...19853928599
[microflora in swine slurry as a parameter in determining the efficiency of deodorizing treatment. i. continuous and discontinuous aeration treatment].liquid manure from the pig, fresh and stored, was treated by continuous and discontinuous aeration. the purpose was to achieve a biologically stabilized and deodorized organic material. the samples which were aerated intermittently had good decomposition rates for nitrogen compounds and even so good deodorization effects. in spite of a minimum of dissolved oxygen of appr. 0.2 mg/l, good deodorization could be achieved by intermittent aeration for 23 days. simultaneous processes of nitrification ...19854050182
mechanism and stereospecificity of the parabactin-mediated iron-transport system in paracoccus denitrificans.the mechanism by which paracoccus denitrificans utilizes parabactin in its iron-transport apparatus is examined. the cellular uptake of 55fe-parabactin, its enantiomer, as well as a large number of its homologues and analogues are measured. furthermore, the ability of these catecholamide ligands to stimulate microbial growth is also determined. the results of these studies point out several structural boundary conditions which the microorganism sets when utilizing iron chelates.19854008483
denitrification of high concentrations of nitrites and nitrates in synthetic medium with different sources of organic carbon. iii. methanol.the denitrification of nitrites and nitrates (1000 mg n/l) in medium containing methanol as a source of organic carbon was studied. continuous cultures of mixed population of autochtonic microflora from bottom sludge of nitrogenous wastewater reservoir were set up in a chemostat-type column and packed bed reactor. the efficiency of denitrification of nitrates in packed bed reactor was 506.7 mg n/l/h whereas denitrification of nitrites was from 8.7 to 16.0 mg n/l/h depending on the granulation of ...19852412408
quantitative selection of denitrifying bacteria in continuous cultures and requirement for organic carbon. i. starch.a mixed population of bacteria from bottom sludge of nitrogen wastewater reservoir was incubated in continuous culture in medium containing 1000 mg nitrate nitrogen/l and starch. maximal efficiency of denitrification was 5 mg n/l/h. marked changes in participation of denitrifying bacteria (16-76%) among total number of bacteria was observed, this being dependent on the ratio of starch concentration (cs) to nitrogen concentration (cn) in the medium. the optimal cs/cn ratio ensuring highest partic ...19852579531
an inducible periplasmic blue copper protein from paracoccus denitrificans. purification, properties, and physiological role.when grown on methylamine as a sole carbon source, paracoccus denitrificans synthesizes a type i blue copper protein which mediates electron transfer between methylamine dehydrogenase and cytochrome c. this blue copper protein does not serve as an electron acceptor for methanol dehydrogenase and is not synthesized by cells grown on methanol or succinate. the blue copper protein and methylamine dehydrogenase were localized in the periplasm of p. denitrificans, whereas formate dehydrogenase was cy ...19852997215
helical packing in the hydrophobic sector of cytochrome c oxidase.an arrangement for the membrane-spanning segments of the three larger subunits of cytochrome c oxidase is proposed on the basis of sequence comparison and polarity distribution estimated from the data available for 11 different organisms.19852991455
cloning of paracoccus cytochrome c oxidase subunit ii.cytochrome c oxidase from paracoccus denitrificans is composed of two subunits, yet is active in both electron transport and proton translocation. a cloning approach and immunologic screening protocol is described for the isolation of the subunit ii gene expressed in e. coli. dna sequencing should establish the extent of homology to eukaryotic oxidase.19852991456
spectroscopic and functional properties of subunit iii-depleted cytochrome oxidase.beef heart cytochrome c oxidase has been depleted of subunit iii by treatment with chymotrypsin. the removal of subunit iii has been evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel fluorography of preparations of the oxidase labeled with [14c]dicyclohexylcarbodiimide prior to proteolysis. removal of subunit iii resulted in a perturbation of the visible spectrum of reduced cytochrome oxidase. subunit iii-depleted oxidase is spectroscopically very similar to the oxid ...19852982875
a comparative study of the resonance raman spectra of bacterial cytochromes.resonance raman spectra were obtained for two newly isolated bacterial cytochromes, alcaligenes faecalis (atcc 8750) c554 and alcaligenes faecalis c556. their spectra were compared with those of mammalian cytochrome c and two other bacterial cytochromes, paracoccus denitrificans c550 and pseudomonas aeruginosa c551. the positions of the raman bands indicated that, while al. c554 and al. c556 were c-type cytochromes with two thioether linkages, several common features found in their raman spectra ...19852982314
osmotic effects on bacterial transport and energetics.paracoccus denitrificans suspended in media containing 20-300 mm nacl swelled progressively as the salt concentration was decreased. the increase in intracellular water volume was accompanied by an enhancement of respiration and a stimulation of the rates of net potassium and alpha-aminoisobutyric acid accumulation. it is postulated that influx of water and consequent lowering of intracellular solute concentration trigger transport mechanisms which are destined to restore the original ion and me ...19852991018
methanol dissimilation in xanthobacter h4-14: activities, induction and comparison to pseudomonas am1 and paracoccus denitrificans.methanol dissimilatory enzymes detected in the methanol autotroph xanthobacter h4-14 were a typical phenazine methosulphate-linked methanol dehydrogenase, a nad+-linked formate dehydrogenase, and a dye-linked formaldehyde dehydrogenase that could be assayed only by activity stains of polyacrylamide gels. this same methanol dehydrogenase activity was found in ethanol-grown cells and was apparently utilized for ethanol oxidation. formaldehyde dehydrogenase activities were investigated in paracoccu ...19852933486
isolation of ubiquinol oxidase from paracoccus denitrificans and resolution into cytochrome bc1 and cytochrome c-aa3 complexes.an enzyme complex with ubiquinol-cytochrome c oxidoreductase, cytochrome c oxidase, and ubiquinol oxidase activities was purified from a detergent extract of the plasma membrane of aerobically grown paracoccus denitrificans. this ubiquinol oxidase consists of seven polypeptides and contains two b cytochromes, cytochrome c1, cytochrome aa3, and a previously unreported c-type cytochrome. this c-type cytochrome has an apparent mr of 22,000 and an alpha absorption maximum at 552 nm. retention of thi ...19852982819
dimeric porin from paracoccus denitrificans.paracoccus denitrificans was shown to contain a 33,000-dalton porin, which produced pores of large (1.6 to 1.8 nm) diameter. cross-linking studies showed that the porin existed as dimers in the outer membrane.19852984184
helix movements and the reconstruction of the haem pocket during the evolution of the cytochrome c family.analysis of cytochromes c (tuna), c2 (rhodospirillum rubrum), c550 (paracoccus denitrificans) and c551 (pseudomonas aeruginosa) shows that they contain 48 residues identifiable as homologous from superposition of the structures. the other 34 to 64 residues are in loops that vary greatly in sequence, length and conformation, or in alpha-helices that are found in only some of the structures. of the 48 homologous residues, 17 are in three segments which pack onto the haem faces. in all four structu ...19852987508
the amino acid sequence of the cytochrome c-554(547) from the chemolithotrophic bacterium thiobacillus neapolitanus.an amino acid sequence is proposed for the cytochrome c-554(547) from the bacterium thiobacillus neapolitanus n.c.i.b. 8539). it consists of a polypeptide chain of 91 residues, with a pair of haem-attachment cysteine residues at positions 15 and 18. there is similarity in sequence with each of the halves of the sequence of the dihaem cytochromes c4 and with a cytochrome c-554(548) from a halophilic strain of paracoccus. detailed evidence for the amino acid sequence of the protein has been deposi ...19852988504
complexity in the redox titration of the dihaem cytochrome c4.redox titration of the dihaem, two domain cytochromes c4 from pseudomonas aeruginosa, pseudomonas stutzeri and azotobacter vinelandii showed complex behaviour indicative of the presence of two redox components. in the case of the p. stutzeri cytochrome c4, two spectroscopically distinct components were present during the redox titration. in contrast, cytochrome c-554(548) from a halophilic paracoccus species is a stable dimer of a monohaem cytochrome which shows close homology to cytochrome c4, ...19852990552
the role of subunit iii in bovine cytochrome c oxidase. comparison between native, subunit iii-depleted and paracoccus denitrificans enzymes.in order to obtain information on the role of subunit iii in the function and aggregation state of cytochrome c oxidase, the kinetics of ferrocytochrome c oxidation by the bovine cytochrome c oxidase depleted of its subunit iii were studied and compared with those of the oxidase isolated from p. denitrificans which contains only two subunits. the aggregation state of both enzymes dispersed in dodecyl maltoside was also compared. the two-subunit oxidase from p. denitrificans gave linear eadie-hof ...19852990554
partial purification and characterization of glutaryl-coenzyme a dehydrogenase, electron transfer flavoprotein, and electron transfer flavoprotein-q oxidoreductase from paracoccus denitrificans.glutaryl-coenzyme a (coa) dehydrogenase and the electron transfer flavoprotein (etf) of paracoccus denitrificans were purified to homogeneity from cells grown with glutaric acid as the carbon source. glutaryl-coa dehydrogenase had a molecular weight of 180,000 and was made up of four identical subunits with molecular weights of about 43,000 each of which contained one flavin adenine dinucleotide molecule. the enzyme catalyzed an oxidative decarboxylation of glutaryl-coa to crotonyl-coa, was maxi ...19852991202
monoclonal antibodies to cytochrome c from paracoccus denitrificans: effects on electron transport reactions.the effect of a monoclonal antibody to a soluble cytochrome c from paracoccus denitrificans was tested on the membrane-bound electron-transport system of this bacterium. this antibody (f3-10.2) and one previously described (f3-29.4) (kuo, l.m., davies, h.c. and smith, l. (1984) biochim. biophys. acta 766, 472-482) were deduced to bind to the cytochrome c in the area including amino acid residue number 23 on a loop on the side of the heme crevice. in contrast to the observations with the previous ...19852994723
phylogenetic origins of the plant mitochondrion based on a comparative analysis of 5s ribosomal rna sequences.the complete nucleotide sequences of 5s ribosomal rnas from rhodocyclus gelatinosa, rhodobacter sphaeroides, and pseudomonas cepacia were determined. comparisons of these 5s rna sequences show that rather than being phylogenetically related to one another, the two photosynthetic bacterial 5s rna sequences show that rather than being phylogenetically related to one another, the two photosynthetic bacterial 5s rnas share more sequence and signature homology with the rnas of two nonphotosynthetic ...198511542018
the phylogeny of purple bacteria: the alpha subdivision.the technique of oligonucleotide cataloging shows the purple photosynthetic eubacteria to comprise three major subdivisions, temporarily called alpha, beta, and gamma--previously designated groups i-iii by gibson et al. (1979). each subdivision contains a number of non-photosynthetic genera in addition to the photosynthetic ones. the alpha subdivision, the subject of the present report, contains most but not all of the species that fall into the classically defined genera rhodospirillum, rhodo ...198411541974
electron transport in paracoccus halodenitrificans and the role of ubiquinone.the membrane-bound nadh oxidase of paracoccus halodenitrificans was inhibited by dicoumarol, 2-n-heptyl-4-hydroxy-quinoline-n-oxide (hqno), and exposure to ultraviolet light (at 366 nm). when the membranes were extracted with n-pentane, nadh oxidase activity was lost. partial restoration was achieved by adding the ubiquinone fraction extracted from the membranes. succinate oxidation was not inhibited by dicoumarol or hqno, but was affected by ultraviolet irradiation or n-pentane extraction. ...198411536581
the growth of paracoccus halodenitrificans in a defined medium.a synthetic medium, consisting of inorganic salts and any of a number of carbon sources, supported the aerobic growth of paracoccus halodenitrificans when supplemented with thiamine. the same medium plus an appropriate nitrogenous oxide supported anaerobic growth when additionally supplemented with methionine. the observation that vitamin b12 or betaine replaced methionine suggested that p. halodenitrificans had a defect in the cobalamin-dependent pathway for methionine biosynthesis, as well a ...198411536589
1h nmr spectroscopy of paracoccus denitrificans cytochrome c-550.the 1h nmr spectra of ferri- and ferro-cytochrome c-550 from paracoccus denitrificans (atcc 13543) have been investigated at 300 mhz. the ferri-cytochrome c-550 shows hyperfine-shifted heme methyl resonances at 29.90, 29.10, 16.70, and 12.95 ppm and a ligand methionyl methyl resonance at -15.80 ppm (ph 8 and 23 degrees c). four ph-linked structural transitions were detected in spectra taken as a function of ph. the transitions have been interpreted as loss of the histidine heme ligand (pk less t ...19846087896
eubacteria have 3 growth modes keyed to nutrient flow. consequences for the concept of maintenance and maximal growth yield.aerobic growth of escherichia coli and paracoccus denitrificans has been studied in chemostat, fed batch, and recycling fermentor modes under carbon and energy limitation. two abrupt drops or discontinuities in molar growth yield, y, have been found that occur over relatively short ranges in the value of specific growth rate. before the first discontinuity, y is constant and maximal. after the first discontinuity, at a doubling time of 33 h, y becomes constant again and independent of mu until t ...19846202266
effects of monoclonal antibodies to bovine and paracoccus denitrificans cytochromes c on reactions with oxidase, reductase and peroxidase.the effects of monoclonal antibodies to bovine and paracoccus denitrificans cytochromes c (kuo, l.m. and davies, h.c. (1983) mol. immunol. 20, 827-838) in the reactions of the cytochromes c with cytochrome c oxidase, reductase and peroxidase were studied. spectrophotometric assays were employed, under conditions where binding of cytochrome c to the enzymes appears to be rate-limiting. less than stoichiometric amounts of antibodies to p. denitrificans cytochrome c added to the cytochrome rendered ...19846205693
patterns of product inhibition for bacterial nitrite reductase.product inhibition has been examined in the turnover kinetics of cytochrome cd1 from pseudomonas aeruginosa (atcc 19429) and from paracoccus denitrificans1 (atcc 13456). a common characteristic was a decrease in rate during the time course of assays that was not due to substrate depletion or irreversible inactivation. the product of nitrite reduction, nitric oxide (no), acted as a product inhibitor in anaerobic assays with an apparent ki of 0.2 microm, but only if the enzyme was first preincubat ...19846091654
the antibacterial action of tinopal an.the bactericidal activity of tinopal an [1,1-bis(3,n-5-dimethyl-benzoxazol-2-yl)-methine p-toluene sulphonate] was shown to be due to a mechanism entirely independent of its inhibitory effects upon nadh dehydrogenase which were reported previously. whereas the compound had no significant effect upon dna synthesis in escherichia coli d22, rna and protein synthesis were immediately and markedly inhibited. in confirmation, tinopal an caused an immediate cessation in inducible beta-galactosidase syn ...19846206200
the effect of oxygen on denitrification during steady-state growth of paracoccus halodenitrificans.steady-state cultures of paracoccus halodenitrificans were grown anaerobically prior to establishing steady states at different concentrations of oxygen. in the absence of oxygen, nitrate-limited cultures produced dinitrogen, and as the oxygen supply increased, these cultures produced nitrous oxide, then nitrite. these changes reflected two phenomena: the inactivation of nitrous oxide reductase by oxygen and the diversion of electrons from nitrite to oxygen.19846712418
on the effects of thiocyanate and venturicidin on respiration-driven proton translocation in paracoccus denitrificans.a fast-responding o2 electrode has been used to confirm and extend observations of a significant kinetic discrepancy between o2 reduction and consequent proton translocation in 'o2-pulse' experiments in intact cells of p. denitrificans. the permeant, chaotropic scn- ion abolishes this discrepancy, and greatly increases the observable----h+/o ratio, to a value approaching its accepted, true, limiting stoichiometry. the observable h+ decay rates are very slow, particularly in the absence of scn-. ...19846743650
isolation and characterization of megaplasmid dna from lithoautotrophic bacteria.a method is described for the preparative isolation of megaplasmids ranging in size from 340 to 700 kb. these plasmids were isolated from chemolithoautotrophic bacteria including the species alcaligenes, pseudomonas, and paracoccus. the procedure was based on alkaline sodium dodecyl sulfate lysis of the cells, followed by heat treatment, salt precipitation, several phenol extractions, dialysis steps, and proteinase and rnase treatment. the various parameters were evaluated and controlled. hydrog ...19846098903
electron-transfer reactions of photoreduced flavin analogues with c-type cytochromes: quantitation of steric and electrostatic factors.we have found correlations between rate constants and the difference in redox potential of the reactants for electron-transfer reactions between oxidized cytochromes and either photoproduced riboflavin or flavin mononucleotide (fmn) semiquinones (the latter rate constants extrapolated to infinite ionic strength). the riboflavin-cytochrome rate constants are about 70% of those for reduction by lumiflavin, probably because of steric interference by the ribityl side chain. reduction of cytochromes ...19846093864
bacterial microflora participating in the removal of nitrogen from industrial wastewaters by nitrification and denitrifaction.nitrification and denitrification were used to purify industrial nitrogenous-organic wastewaters containing 1,130 mg n/l (mainly in the form of ammonium and urea) and with mean cod 1,050 mg o2/l. the use of a five-step system consisting of two denitrifying beds and three nitrifying activated sludge reactors gave 99% removal of nitrogen and 54% reduction in cod during total retention time of the wastes of 81 hours. the participation of stage i nitrifying bacteria in the activated sludge was 4.4 t ...19846205553
nitrous oxide reductase and the 120,000 mw copper protein of n2-producing denitrifying bacteria are different entities.zumft and matsubara [1982) febs lett. 148, 107-112) isolated a 120,000 mw copper protein from certain denitrifying bacteria which were capable of producing n2. the presence of this protein correlated with a nutritional requirement of copper for growth on and reduction of n2o by the bacteria. the copper protein was alleged by these workers to be nitrous oxide reductase. however, it is shown that the copper protein and nitrous oxide reductase have different molecular weights and exhibit different ...19846712645
heterotrophic nitrification among denitrifiers.twelve denitrifying bacteria representing six genera were tested for an ability to nitrify pyruvic oxime heterotrophically. six of these bacteria exhibited appreciable nitrification activity, yielding as much as 5.8 mm nitrite and little or no nitrate when grown in a mineral salts medium containing 7 mm pyruvic oxime and 0.05% yeast extract. of the six active bacteria, four (pseudomonas denitrificans, pseudomonas aeruginosa, and two strains of pseudomonas fluorescens) could grow on yeast extract ...19846721486
denitrification and nitrite reduction: pseudomonas aeruginosa nitrite-reductase.present knowledge of the different enzymatic steps of the denitrification chains in various bacteria, particularly paracoccus denitrificans and pseudomonas aeruginosa has been briefly reviewed. the question whether nitric oxide (no), nitrous oxide (n2o) and other nitrogen derivatives are obligatory intermediates has been discussed. the second part is an extensive review of the structure and the function of a key enzyme in denitrification, cytochrome c551-nitrite-oxidoreductase from p. aeruginosa ...19846331530
proposed structure for the noncovalently associated heme prosthetic group of dissimilatory nitrite reductases. identification of substituents.the substituents of the noncovalently associated heme prosthetic group of the bacterial nitrite reductase-cytochrome oxidase (ec 1.9.6.1 or ec 1.9.3.2.) from pseudomonas aeruginosa (atcc 19429) and paracoccus denitrificans (atcc 13456) have been identified. this was accomplished by 1h nmr, infrared, visible, and mass spectroscopic techniques applied to semi-purified heme and purified methyl ester derivatives of the iron-free porphyrin. the main structural features are as follows. 1) the macrocyc ...19846420411
a dissimilatory nitrite reductase in paracoccus halodenitrificans.paracoccus halodenitrificans produced a membrane-associated nitrite reductase. spectrophotometric analysis showed it to be associated with a cd-cytochrome and located on the inner side of the cytoplasmic membrane. when supplied with nitrite, membrane preparations produced nitrous oxide and nitric oxide in different ratios depending on the electron donor employed. the nitrite reductase was maximally active at relatively low concentrations of sodium chloride and remained attached to the membranes ...19846712419
immunochemical identification of a two-subunit nadh-ubiquinone oxidoreductase from paracoccus denitrificans.analysis by crossed-immunoelectrophoresis of paracoccus denitrificans membrane vesicles has shown that only one antigen stains for nadh dehydrogenase activity. this activity could be partially purified by a combination of gel filtration and ion-exchange chromatography of membrane vesicles that had been solubilised in the non-ionic detergent nonidet p-40. from the limited number of precipitates observed after crossed immunoelectrophoresis of this partially purified preparation of nadh dehydrogena ...19846434308
saturation behavior of the manganese-containing superoxide dismutase from paracoccus denitrificans.a pulse radiolysis study of the mn-superoxide dismutase from paracoccus denitrificans has shown that, at concentration of 0(2)-. below 0.8 x 10(-4)m, the catalyzed dismutation of 0(2)-. is a first order reaction with regard to 0(2)-.. at concentration of 0(2)-. above 0.8 x 10(-4)m, the mn-superoxide dismutase is shown to catalyze superoxide dismutation with a mechanism which exhibits saturation kinetics. this behavior was previously found in the bovine cu/zn-superoxide dismutase and in the fe-su ...19836860328
turnover of cytochrome c oxidase from paracoccus denitrificans.the heme aa3 type cytochrome oxidase from paracoccus denitrificans incorporated into vesicles with phospholipid reacts during turnover much as the oxidase from mitochondria does. the spectrophotometric changes observed at various wavelengths are closely similar, and the rate is about one-half of that for beef heart oxidase under the same conditions. the rate of appearance of oxidized cytochrome c on initiation of the reaction is also similar and depends on the previous treatment of the oxidase a ...19836296100
the distribution of redox equivalents in the anaerobic respiratory chain of paracoccus denitrificans. 19836297904
electrochemically driven respiration in mitochondria and paracoccus denitrificans. the coupling of the electrochemistry of horse heart cytochrome c with respiration in mitochondria and a model thereof, paracoccus denitrificans.by exploiting the rapid, direct electrochemistry of horse heart cytochrome c at a modified gold electrode it has been possible to couple the electrode reaction with respiration in rat liver mitochondria and in protoplasts of paracoccus denitrificans, but not in protoplasts from e. coli. oxidation of endogenous and exogenous sources of reducing equivalents via cytochrome c is also observed.19836299804
microbial growth associated with granular activated carbon in a pilot water treatment facility.the microbial dynamics associated with granular activated carbon (gac) in a pilot water treatment plant were investigated over a period of 16 months. microbial populations were monitored in the influent and effluent waters and on the gac particles by means of total plate counts and atp assays. microbial populations between the influent and effluent waters of the gac columns generally increased, indicating microbial growth. the dominant genera of microorganisms isolated from interstitial waters a ...19836625567
presence of a new cytochrome b - like pigment with a peak at 567 nm in various aerobic bacteria.several physiological groups of bacteria were examined for the presence of a cytochrome b - like pigment which is demonstrable in dithionite-reduced minus substrate-reduced difference spectra. this pigment is characterized by an unusually high alpha band at 567 nm, a low concentration relative to conventional cytochromes, and an inability to be fully reduced by endogenous substrates or nadh. previous studies with one denitrifying and nondenitrifying species of the genus pseudomonas, in paracoccu ...19836652580
the function of cytoplasmic membrane of paracoccus denitrificans in controlling the rate of reduction of terminal acceptors.the rate of reduction of terminal acceptors (nitrate, nitrite, and oxygen) in anaerobically grown cells of paracoccus denitrificans increased on permeabilization of cytoplasmic membrane. it was proved that under aerobic conditions the increase of the rate of nitrate reduction was caused by: (i) the abolishment of the permeability barrier for nitrate, (ii) the enhancement of the influx of redox equivalents to the respiratory chain due to the stimulation of succinate dehydrogenase reaction, and (i ...19836684550
immunochemical relationship of the major outer membrane protein of rhodopseudomonas sphaeroides 2.4.1 to proteins of other photosynthetic bacteria.immunoblots of sodium dodecyl sulfate-polyacrylamide gels derived from outer membrane preparations of various strains of rhodopseudomonas sphaeroides revealed polypeptides which cross-reacted with antibody directed against the major outer membrane protein of r. sphaeroides 2.4.1. immunochemical quantitation of the major outer membrane protein of strain 2.4.1 showed approximately 5.5 x 10(4) molecules per cell whether cells were grown chemoheterotrophically or photoheterotrophically. rhodospirill ...19836188744
effect of various sources of organic carbon and high nitrite and nitrate concentrations on the selection of denitrifying bacteria. ii. continuous cultures in packed bed reactors.the effect of different organic compounds, nitrites and nitrates at the concentration of 1,000 mg n/l on the quantitative and strain-specific selection of denitrifying bacteria was determined in anaerobic packed bed reactors. both the source of carbon and nitrogen form influenced strain specificity and the frequency of occurrence of denitrifying bacteria. the frequency of denitrifying bacteria within packed bed reactor ranged in different media from 11% (glucose and nitrates) to 100% (methanol a ...19836194668
the effect of uncoupler on the distribution of the electron flow between the terminal acceptors oxygen and nitrite in the cells of paracoccus denitrificans.the preferential utilization of oxygen, the terminal acceptor, in anaerobically grown cells of paracoccus denitrificans was abolished in the presence of uncoupler (3 microm carbonyl cyanide m-chlorophenylhydrazone) which brought about a switch to the reduction of nitrite. it has been proved by measuring the redox state of cytochromes that this effect is due to the inhibition of the electron flow to oxygen caused by nitrite, which attains the site of its inhibitory action when the membrane potent ...19836661222
cytochrome c1 from paracoccus denitrificans.cytochrome c1 was purified from the bacterium paracoccus denitrificans. it is an acidic, hydrophobic polypeptide with an apparent molecular weight of around 65000 and a single, covalently attached heme; it cross-reacts immunologically with cytochrome c1 from yeast mitochondria. the amino acid sequence of the tryptic heme peptide of the bacterial cytochrome c1 shows extensive homology to the corresponding region of beef heart cytochrome c1 [wakabayashi, s. et al. (1982) j. biol. chem. 257, 9335-9 ...19836319128
characterisation of membrane vesicles from paracoccus denitrificans and measurements of the effect of partial uncoupling on their thermodynamics of oxidative phosphorylation.1. vesicles from paracoccus denitrificans were prepared by applying an osmotic shock to spheroplasts derived from cells that had been grown anaerobically with succinate as carbon source and nitrate as electron acceptor. in the presence of either phenazinemethosulphate or n,n,n' n',-tetramethyl-p-phenylenediamine, the oxidation of isoascorbate supported the uptake of both s14cn- and 86rb+ (in the presence of valinomycin), whereas nadh and succinate oxidation resulted only in s14cn- uptake. these ...19836301833
the effects of partial uncoupling upon the kinetics of atp synthesis by vesicles from paracoccus denitrificans and by bovine heart submitochondrial particles. implications for the mechanism of the proton-translocating atp synthase.1. reduction in the magnitude of the respiration-dependent protonmotive force (proton electrochemical gradient in mv) of vesicles from paracoccus denitrificans, and of submitochondrial particles, has been found to be paralleled small increases in s50% values for both adp and pi. for example, reduction of the protonmotive force of p. denitrificans vesicles from 145 mv to 110 mv was accompanied by an increase of s50% (adp) from 8 microm to 18 microm, and an increase of s50% (pi) from 0.33 mm to 1. ...19836301834
production, isolation and characterization of monoclonal antibodies to cytochromes c of beef heart and paracoccus denitrificans.hybridoma cell lines secreting monoclonal antibodies which bind beef heart cytochrome c or paracoccus denitrificans cytochrome c have been produced using spleen cells from balb/c mice immunized with cytochrome c. immunization was performed with either the native cytochrome c, succinylated hemocyanin-conjugated cytochrome c, or beef heart cytochrome c polymerized with glutaraldehyde. of 10 such fusions, the hybridization frequency ranged from 0 to 42%. the cell fusion efficiency, the possible fac ...19836312296
reaction of cytochrome c in the electron-transport chain of paracoccus denitrificans.the reaction of the cytochrome c oxidase (ferrocytochrome c:oxygen oxidoreductase, ec 1.9.3.1) of paracoccus denitrificans cytoplasmic membranes with the endogenous cytochrome c of the membranes was studied, as well as its interaction with added exogenous cytochrome c from p. denitrificans or bovine heart. the polarographic method was employed, using n,n,n',n'-tetramethyl-p-phenylenediamine plus ascorbate to reduce the cytochrome c. we found that overall electron transport can proceed maximally ...19836315059
atp-amp phosphotransferase from paracoccus denitrificans.adenylate kinase has been extracted from paracoccus denitrificans by toluene treatment and purified 370-fold (overall yield of 44%, 554 u/mg, mr 22 000, pi 4.7; ph optimum 7.5-8.0) by using successive column chromatography on deae-cellulose de-52, matrex-blue a and sephadex g-75. the enzyme was homogeneous by dodecylsulfate gel electrophoresis and constancy of specific activity across the single peak found in sephadex g-75 chromatography. crystals in the form of heavy needles have been obtained ...19836315432
consensus structure and evolution of 5s rrna.a consensus structure model of 5s rrna presenting all conserved nucleotides in fixed positions has been deduced from the primary and secondary structure of 71 eubacterial, archaebacterial, eukaryotic cytosolic and organellar molecules. phylogenetically related groups of molecules are characterized by nucleotide deletions in helices iii, iv and v, and by potential base pair interactions in helix iv. the group-specific deletions are correlated with the early branching pattern of a dendrogram calcu ...19836835839
phosphate transport system in paracoccus denitrificans.pi uptake in cells or spheroplasts of paracoccus denitrificans is biphasic; only the first rapid phase represents net pi transport. the second phase is limited by the rate of pi utilization inside the cell, i.e., mainly by its esterification, and as such it was inhibited by dccd. the pi/dicarboxylate antiporter does not seem to be operative, and its inhibitor n-butylmalonate did not exert specific inhibition. pi transport is inhibited by sh reagents; the most potent inhibitor is pcmb, and mersal ...19836853472
15n-tracer and nmr studies on the pathway of denitrification. evidence against trioxodinitrate but for nitroxyl as an intermediate.the effects of four species of denitrifying bacteria on the conversion of [15n]nitrite to trioxodinitrate (hn2o3-) and n2o and of trioxodinitrate to n2o were studied. for all species, the n2o produced in the presence of [15n]nitrite and trioxodinitrate was isotopically randomized throughout the period of incubation and was not composed at the outset predominantly of 14n2o or 14n2o plus 15n2o. the n2o produced was also heavily enriched in 15n at times when the trioxodinitrate pool was only weakly ...19836833215
dicyclohexylcarbodiimide does not inhibit proton pumping by cytochrome c oxidase of paracoccus denitrificans.the effect of dicyclohexylcarbodiimide (dccd) on the proton pumping two-subunit cytochrome c oxidase from paracoccus denitrificans was investigated. purified paracoccus oxidase was reconstituted into phospholipid vesicles by cholate dialysis. following incubation with increasing amounts of dccd, proton ejection was recorded in response to reductant pulses with reduced cytochrome c. concentrations of dccd which greatly reduced proton pumping by bovine cytochrome c oxidase used as a control were f ...19836305656
receptor sites involved in posttranslational transport of apocytochrome c into mitochondria: specificity, affinity, and number of sites.assembly of cytochrome c involves a series of steps: synthesis of apocytochrome c on free ribosomes, specific binding of apocytochrome c to the mitochondrial surface, transfer across the outer membrane, covalent addition of protoheme, refolding of the polypeptide chain, and association of holocytochrome c with its functional sites at the inner membrane. the binding step of apocytochrome c to neurospora crassa mitochondria was studied by inhibiting the subsequent transfer steps with the heme anal ...19836308663
proton translocation during denitrification by a nitrifying--denitrifying alcaligenes sp.a heterotrophic nitrifying alcaligenes sp. from soil was grown as a denitrifier on nitrate and subjected to oxidant pulse experiments to ascertain the apparent efficiencies of proton translocations during o2 and nitrogen-oxide respirations. with endogenous substrate as the reducing agent the leads to h+/2e- ratios, extrapolated to zero amount of oxidant per pulse, were 9.4, 3.7, 4.3 and 3.5 for o2, nitrate, nitrite and n2o, respectively. the value for o2 and those for the n-oxides are, respectiv ...19836311094
sequence homology of bacterial and mitochondrial cytochrome c oxidases. partial sequence data of cytochrome c oxidase from paracoccus denitrificans.the aerobic electron transport chain of paracoccus denitrificans is very similar to that of mitochondria. it has therefore been suggested that this bacterium might be evolutionarily related to mitochondria. the two subunits (mr 45.000 and 28.000) of the paracoccus cytochrome c oxidase were isolated and partially sequenced. the sequences were found to be surprisingly homologous to sequences of the subunits i and ii of mitochondrial cytochrome c oxidases. the data provide a molecular basis for the ...19836315007
comparison of denitrification by pseudomonas stutzeri, pseudomonas aeruginosa, and paracoccus denitrificans.a comparison was made of denitrification by pseudomonas stutzeri, pseudomonas aeruginosa, and paracoccus denitrificans. although all three organisms reduced nitrate to dinitrogen gas, they did so at different rates and accumulated different kinds and amounts of intermediates. their rates of anaerobic growth on nitrate varied about 1.5-fold; concomitant gas production varied more than 8-fold. cell yields from nitrate varied threefold. rates of gas production by resting cells incubated with nitrat ...19836407395
proton-coupled energy transduction by biological membranes. principles, pathways and praxis. 19827183456
isolation of paracoccus denitrificans cytochrome cd1: comparative kinetics with other nitrite reductases. 19826284044
changes in the cytochrome c content during the aerobic adaptation of paracoccus denitrificans.concentration of cytochrome c decreases when shaking anaerobically grown cells in a non-growth medium down to 50% of the original amount in the cell, depending on the degree of aeration. only 10-20% of this amount can be found in the adaptation medium. the main portion of cytochrome c is degraded during the adaptation. inhibitors of proteinases do not influence the degradation. addition of mammalian cytochrome c fully prevents the degradation of bacterial cytochrome c but not its release from ce ...19826277754
15n,18o tracer studies on the activation of nitrite by denitrifying bacteria. nitrite/water-oxygen exchange and nitrosation reactions as indicators of electrophilic catalysis. 19826806292
alterations in the phospholipid composition of rhodopseudomonas sphaeroides and other bacteria induced by tris.alterations in the phospholipid head group composition of most strains of rhodopseudomonas sphaeroides, as well as rhodopseudomonas capsulata and paracoccus denitrificans, occurred when cells were grown in medium supplemented with tris. growth of r. sphaeroides m29-5 in tris-supplemented medium resulted in the accumulation of n-acylphosphatidylserine (naps) to as much as 40% of the total whole-cell phospholipid, whereas naps represented approximately 28 an 33% of the total phospholipid when r. c ...19826982264
oxygen tolerance of strictly aerobic hydrogen-oxidizing bacteria.growth of various bacteria, especially aerobic hydrogen-oxidizing bacteria, in the presence of 2 to 100% (v/v) oxygen in the gas atmosphere was evaluated. the bacterial strains included alcaligenes eutrophus, a. paradoxus, aquaspirillum autotrophicum, arthrobacter spec. strain 11 x, escherichia coli, arthrobacter globiformis, nocardia opaca, n. autotrophica, paracoccus denitrificans, pseudomonas facilis, p. putida, and xanthobacter autotrophicus. under heterotrophic conditions with fructose or g ...19827049081
isolation and characterization of a protein with cyanide-sensitive superoxide dismutase activity from the prokaryote, paracoccus denitrificans.1. a protein with cyanide-sensitive superoxide dismutase activity was isolated from the prokaryote paracoccus denitrificans. 2. this enzyme, present in low amount in the cell, represented not more than 10% of the total cellular superoxide dismutase activity. it was obtained in a form which was 20-40-times less active than the main superoxide dismutase of p. denitrificans which is a manganese-containing enzyme. 3. it was a soluble monomeric enzyme, highly negatively charged (pi = 4.8), with an ap ...19827066332
a novel inhibitor of nadh dehydrogenase in paracoccus denitrificans. 19827084467
the 5s ribosomal rnas of paracoccus denitrificans and prochloron.the nucleotide sequences of the 5s rrnas of paracoccus denitrificans and prochloron sp. are (formula: see text), respectively. specific phylogenetic relationships of p. denitrificans with purple non-sulphur bacteria, and of prochloron with cyanobacteria are demonstrated, and unique features of potential secondary structure are described.19827099971
proton translocation and proline uptake associated with reduction of nitric oxide by denitrifying paracoccus denitrificans. 19827138549
tetraphenylborate-sensitive electrode for measuring membrane potential.the paper describes the construction of a new type of ion-selective electrode sensitive to tetraphenylborate (tpb-) and its electric characteristics. the electrode responds to increasing concentrations of the tpb- anion in accordance with the nernst equation and can be used down to 0.1 microm concentration. the applicability of the electrode for measuring the membrane potential (positive inside) was proved in inside-out oriented membrane vesicles derived from paracoccus denitrificans. the calcul ...19827173748
benzyl viologen cation radical: first example of a perfectly selective anion ionophore of the carrier type. 19827181886
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