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a pulse-radiolysis study of the catalytic mechanism of the iron-containing superoxide dismutase from photobacterium leiognathi.the mechanism of the enzymic reaction of an iron-containing superoxide dismutase purified from the marine bacterium photobacterium leiognathi was studied by using pulse radiolysis. measurements of activity were done with two different preparations of enzyme containing either 1.6 or 1.15 g-atom of iron/mol. in both cases, identical values of the second-order rate constant for reaction between superoxide dismutase and the superoxide ion in the ph range 6.2-9.0 (k=5.5 x 10(8) m-1-s-1 at ph 8.0) wer ...197715540
interaction of the mannosephilic lectins of pseudomonas aeruginosa with luminous species of marine enterobacteria.the marine bacteria beneckea harveyi and photobacterium leiognathi were shown to bear mannose-containing binding sites for the mannosephilic lectins of pseudomonas aeruginosa and concanavalin a (con a). the interaction between the lectins and the marine bacteria was demonstrated by the bacteriagglutination test, by adsorption of the lectins onto the bacteria and by mannose-specific peroxidase-binding to the lectin-coated bacteria. treatment of the bacteria with formaldehyde, phenol, ethanol or b ...1979120927
superoxide dismutases: defence against endogenous superoxide radical.attempts to measure the rate of o2- production, in whole cells or in intact subcellular organelles, are frustrated by the endogenous superoxide dismutase (sod). streptococcus faecalis contains a single manganese-sod which was isolated and used as an antigen in the rabbit. a precipitating and inhibiting antibody was obtained and used to suppress the sod in crude lysates of s. faecalis. it allowed the demonstration that 17% of the total oxygen uptake by such lysates, in the presence of nadh, was a ...1978225147
a calorimetric investigation of the growth of the luminescent bacteria beneckea harveyi and photobacterium leiognathi.direct calorimetric determinations of the rate of heat production along with simultaneous determinations of the rate of photon emission and the number of viable cells have provided insight into the growth of beneckea harveyi and photobacterium leiognathi. these experiments were performed with a tronac isothermal microcalorimeter modified with a fiber optic light guide to allow in situ detection of light. escherichia coli and a dark variant of p. leiognathi were also examined to provide points of ...1977401656
control of luciferase synthesis in a newly isolated strain of photobacterium leiognathi.in previous studies with luminous bacteria of all different species it has been reported that the synthesis of luciferase is autoinducible: during growth at low cell densities synthesis is effectively repressed while after induction, at higher cell densities, the rate of synthesis of enzyme is up to five times the growth rate. in this paper we report on newly isolated strains of photobacterium leiognathi which show continued luciferase synthesis irrespective of the cell density. the specific syn ...1977603341
the lux genes in photobacterium leiognathi are closely linked with genes corresponding in sequence to riboflavin synthesis genes.three open reading frames (orfs) have been found in the region downstream of the luxg gene in the photobacterium leiognathi lux operon. these genes (orf i, ii, and iii) are not only closely linked to the lux operon and transcribed in the same direction but also show the same organization and code for proteins homologous in sequence to the gene products of ribb, riba, and ribh of bacillus subtilis, respectively. the photobacterium leiognathi gene (orf ii) corresponding to riba was expressed in es ...19921339274
crystallization of photobacterium leiognathi non-fluorescent flavoprotein, an unusual flavoprotein with limited sequence identity to bacterial luciferase.single crystals of the non-fluorescent flavoprotein (nfp) purified from photobacterium leiognathi strain s1 have been grown from ammonium sulphate solutions using the hanging drop vapour diffusion technique. the crystals grow as thin (0.06 mm) plates and belong to the orthorhombic space group c222(1): a = 57.06(3) a, b = 92.41(6) a, c = 99.52(6) a. there is one nfp monomer per asymmetric unit and crystals diffract to 2.2 a spacings on film. a complete native data set to 2.5 a resolution has been ...19921560468
electronic excitation transfer in the complex of lumazine protein with bacterial bioluminescence intermediates.fluorescence dynamics measurements have been made on the bioluminescence reaction intermediates using photobacterium leiognathi, vibrio fischeri, and vibrio harveyi luciferases, both alone and in mixtures with photobacterium phosphoreum lumazine protein. each luciferase produces a "fluorescent transient" intermediate on reaction with the bioluminescence substrates, fmnh2, tetradecanal, and o2, and all have a fluorescence quantum yield about 0.3, with a predominant lifetime around 10 ns. the p. l ...19912069948
[expression of photobacterium leiognathi bioluminescence system genes in escherichia coli].expression of photobacterium leiognathi bioluminescence genes under the control of lac, tac, tet promoters in escherichia coli cells has been studied. the position of the genes for aliphatic aldehyde biosynthesis and for the synthesis of luciferase subunits was identified. the plasmid pbrpl1 has been constructed containing the system of bioluminescence genes devoid of promoter following the polylinker dna fragment. the plasmid can be used for selection of promoter containing dna sequences as wel ...19902185420
isolation of bioluminescent functions from photobacterium leiognathi: analysis of luxa, luxb, luxg and neighboring genes.genes encoding luminescence of photobacterium leiognathi have been cloned in escherichia coli. the luminescent clones were readily apparent. among them, a clone containing a recombinant plasmid with a 13.5-kb insertion was identified. this dna fragment contained all of the luminescence-encoding genes. the luciferase-encoding genes (lux) in this dna fragment were localized. we have sequenced a part of the cloned lux region and identified the luxa, luxb and luxg genes encoding the alpha and beta s ...19902311938
copper-zinc superoxide dismutase of caulobacter crescentus: cloning, sequencing, and mapping of the gene and periplasmic location of the enzyme.although widely found in the cytoplasm of eucaryotes, the copper-zinc form of superoxide dismutase (cuznsod) has been identified in only a small number of bacterial species. one species is the freshwater bacterium caulobacter crescentus, which also contains an sod with iron as the metal cofactor (fesod). to investigate the function of this cuznsod and its structural relationship to the eucaryotic cuznsods, the gene encoding cuznsod (sodc) of c. crescentus cb15 was cloned and sequenced. by hybrid ...19902345128
the nucleotide sequence of a small mett trna operon from photobacterium leiognathi. 19902362826
long-term preservation of active luminous bacteria by lyophilization.a simple method for long-term preservation of luminous bacteria is described. cells of vibrio fischeri, photobacterium leiognathi and four strains of p. phosphoreum were suspended in a protective medium of low ionic strength (1% nacl) supplemented with 15% lactose and 2% soluble starch, and lyophilized. the freeze-dried preparations were sealed under vacuum and stored at 4 degrees c. luminous bacteria were resuscitated after six months by adding 2% nacl up to the original volume. the rehydrated ...19892652990
[comparative restriction analysis of chromosomal dna of strains of photobacterium leiognathi].chromosomal dna in 5 hereditary variants occurring in photobacterium leiognathi population was subjected to restriction analysis. the variants differed in the levels and regulation of luminescence and colony morphology. agarose electrophoresis of dna fragments isolated after exposure to hind ii, bam hi, bgl i and pst i restriction endonucleases revealed respectively 38, 28, 35 and 29 fragments equally distributed by their molecular weights. electrophoregrams of the 5 strains were absolutely iden ...19882837154
nucleotide sequence of part of photobacterium leiognathi lux region. 19883186447
difference between amino acid residues in the metal-ligand environments of iron- and manganese-superoxide dismutases.alignment of the amino acid sequences of the pseudomonas ovalis and photobacterium leiognathi iron-superoxide dismutases (fe-sods) with the known sequences of the manganese-superoxide dismutases (mn-sods) shows that both types of sod are highly homologous (33-53% identity) and share residues for the metal coordination. the amino acid residues that form the environment of the metal ions appear to be also conserved between the fe- and mn-sods, except that the phe-84 and gln-154 in the mn-sods are ...19883382418
[nucleotide sequence of genes for alpha- and beta-subunits of luciferase from photobacterium leiognathi].nucleotide sequence of the photobacterium leiognathi dna containing genes of alpha and beta subunits of luciferase has been determined. we also deduced amino acid sequence and molecular mass of luciferase and localized luciferase genes in the sequenced dna fragment.19883382442
amino acid sequence of iron-superoxide dismutase from pseudomonas ovalis.the amino acid sequence of iron-superoxide dismutase from pseudomonas ovalis was deduced by the analyses of peptides derived from limited hydrolysis of the aminoethylated or pyridylethylated apoprotein with trypsin, staphylococcus aureus v8 protease, and dilute acid hydrolysis. the polypeptide chain contains 195 amino acid residues and has a calculated mr of 21,421. the sequence is highly homologous (65% identity) to the recently published sequence of the iron-superoxide dismutase from photobact ...19873666146
primary structure of cu-zn superoxide dismutase of brassica oleracea proves homology with corresponding enzymes of animals, fungi and prokaryotes.the complete amino-acid sequence of cu-zn superoxide dismutase from white cabbage (brassica oleracea) is reported. the polypeptide chain consists of 151 amino acids and has a molecular mass of 15,604 da. the primary structure of the reduced and s-carboxymethylated protein was determined by automated solid phase sequence analysis of tryptic fragments and peptides obtained by digestion with staphylococcus aureus proteinase v8. the protein shows a free amino terminus as was found for all non-mammal ...19863790249
bacteriocuprein superoxide dismutase of photobacterium leiognathi. isolation and sequence of the gene and evidence for a precursor form.the gene encoding the bacteriocuprein superoxide dismutase from photobacterium leiognathi, american type culture collection strain 25521, was cloned in a puc12 vector and sequenced. the nucleotide sequence predicted a 22-residue leader peptide amino-terminal to the known bacteriocuprein sequence. the expected precursor bacteriocuprein was directly identified in the in vitro translation products of the cloned gene by polyacrylamide gel electrophoresis and automated edman degradation. enzymaticall ...19873805055
identity of the metal ligands in the manganese- and iron-containing superoxide dismutases.alignment of the amino acid sequence of peptides obtained following digestion of photobacterium leiognathi iron superoxide dismutase with the known sequence of bacillus stearothermophilus manganese superoxide dismutase shows that the residues found to form ligands to the manganese are conserved in the iron enzyme. this indicates that the metal ligands in both proteins are identical.19853967757
[cloning and expression of genes of the luminescence system in photobacterium leiognathi].the genes of photobacterium leiognathi luminescence system were cloned in plasmid puc18. escherichia coli cells harboring a recombinant plasmid pphl1 are luminescent. pphl1 contains luciferase genes and genes responsible for aldehyde biosynthesis. the luminescence of escherichia coli is subject to autoinductor regulation similar to the one existing in luminescent bacteria. the 2.7 kb fragment of photobacterium leiognathi dna containing the genes for alpha- and beta-luciferase subunits were clone ...19873683427
the primary structure of iron-superoxide dismutase from photobacterium leiognathi.the complete amino acid sequence of iron-superoxide dismutase from photobacterium leiognathi was determined. the sequence was deduced following characterization of the peptides obtained from tryptic, chymotryptic, and staphylococcus aureus v-8 protease digests of the apoprotein. the amino acid sequence listed below is made up of 193 residues. it is the first complete sequence to be determined for an iron-superoxide dismutase. the iron-superoxide dismutase shows the same order of homology with th ...19873542995
strain variation in bacteriocuprein superoxide dismutase from symbiotic photobacterium leiognathi.photobacterium leiognathi atcc 25521 (the type strain and light-organ symbiont of ponyfish) is one of the few bacteria that produces a copper-zinc superoxide dismutase, termed bacteriocuprein. we enzymologically and immunologically characterized the bacteriocuprein superoxide dismutases in sonicates from the type strain and nine additional strains of p. leiognathi, each isolated from the light organ of a separate ponyfish specimen, representing seven ponyfish species. the results indicate consid ...19863511030
structural identity between the iron- and manganese-containing superoxide dismutases.we have recently reported the first complete amino acid sequence of an iron-containing superoxide dismutase. the iron enzyme is thought to be closely homologous to the manganese-containing superoxide dismutases. the availability of complete amino acid sequence information for four manganese superoxide dismutases and the crystal structures for two iron and two manganese superoxide dismutases prompted us to investigate the degree of homology between the two proteins at various levels. we report th ...19873508288
isolation of the lux genes from photobacterium leiognathi and expression in escherichia coli.genes necessary for luminescence (lux genes) in the marine bacterium photobacterium leiognathi, strain pl721, were isolated and expressed in escherichia coli. a 15-kb fragment obtained from a partial digestion of pl721 dna with hindiii was cloned into the plasmid pacyc184, resulting in the hybrid plasmid psd721. when psd721 was transformed into e. coli ed8654, the resulting transformants were luminous with no additions to the cells, indicating that it contained the structural genes coding for th ...19873308637
[study in fluid media and at low temperature of reaction of bioluminescence emission of photobacterium leiognathi]. 19704987002
superoxide dismutases.superoxide dismutases (ec 1.15.1.1) are metalloenzymes that catalytically scavenge the superoxide radical. they are essential for the aerobic survival of all forms of life. there are three types of superoxide dismutase, containing manganese, iron, or copper and zinc. the copper--zinc type has generally been isolated from eukaryotic cells except for the enzyme for the symbiotic marine bacterium photobacterium leiognathi. the copper--zinc type, from different sources, has a molecular weight of abo ...19806258885
initiation and control of the bioluminescent symbiosis between photobacterium leiognathi and leiognathid fish. 19873304077
the amino-acid sequence of copper/zinc superoxide dismutase from swordfish liver. comparison of copper/zinc superoxide dismutase sequences.the amino acid sequence of copper/zinc superoxide dismutase from swordfish (xiphias gladius) liver has been determined by alignment of the tryptic peptides according to the known sequence of bovine erythrocyte copper/zinc superoxide dismutase. this alignment has resulted in the ligands to the copper (his-47, 49, 76 and 94) and the zinc (his-76, 85, 134 and asp-97) being conserved in all the copper/zinc superoxide dismutases sequenced so far. also conserved in the sequences are the cysteines form ...19846510412
interaction of lectins from gonads and haemolymph of the sea hare aplysia with bacteria.gonads and haemolymph of two mediterranean species of aplysia (a. depilans and a. fasciata) contain lectins. a. depilans gonad lectin is specific for d-galacturonic acid and d-galactosides, while its haemolymph agglutinin binds n-acetylated sugars. a. fasciata gonad lectin is also specific for d-galacturonic acid, but its haemolymph haemagglutinin exhibits heterogenic specificity. both aplysia gonad lectin and haemolymph agglutinins interact with bacteria, including certain escherichia coli stra ...19863091995
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
the primary structure of cu-zn superoxide dismutase from photobacterium leiognathi: evidence for a separate evolution of cu-zn superoxide dismutase in bacteria.the complete amino-acid sequence of the copper-zinc superoxide dismutase of the photobacterium leiognathi was determined. the fragmentation strategy employed included cyanogen bromide cleavage at its methionine residues and the only tryptophan residue. the s-carboxymethylated chain was further cleaved by means of trypsin, in order to obtain overlapping fragments. for sequence determination automated solid or liquid-phase techniques of edman degradation were used. c-terminal amino acids of the en ...19836884993
the stimulation of bioluminescence in photobacterium leiognathi as a potential prescreen for antitumor agents.the stimulation of bioluminescence in photobacterium leiognathi has previously been described as a test for genotoxic compounds. an adaptation of this procedure has been developed which uses a dim variant of p. leiognathi and permits the prescreening of microbial fermentation broths for potential antitumor agents. bioluminescence in this organism was stimulated by compounds which bind to dna or affect dna synthesis. antibiotics with target sites such as protein, cell wall or rna synthesis, did n ...19852999049
copper-zinc superoxide dismutase from caulobacter crescentus cb15. a novel bacteriocuprein form of the enzyme.a bacteriocuprein is a copper- and zinc-containing superoxide dismutase isolated from a bacterium. until recently, the first and only documented bacteriocuprein was that from the marine bacterium photobacterium leiognathi, which lives symbiotically with leiognathid fishes. a new bacteriocuprein has been discovered, purified, and characterized from the free living, non-symbiotic bacterium, caulobacter crescentus cb15. in its native molecular weight, homodimeric subunit structure, specific activit ...19827050107
[cloning and insertion mutagenesis of dna fragment coding for the luminescent system of photobacterium leiognathi].fragments of dna, obtained from the luminescent bacterium photobacterium leiognathi and inserted into the plasmid pbr322, were found to code for the luminescence expressed in e. coli cells. the genetic functions necessary for light production in e. coli are localized on a dna fragment of about 7 kbp. the insertion mutagenesis was used to define the luminescence functions encoded by the hybrid plasmid.19882852774
the complete nucleotide sequence of the lux regulon of vibrio fischeri and the luxabn region of photobacterium leiognathi and the mechanism of control of bacterial bioluminescence.we have determined the complete nucleotide sequence of a 7622 base pair fragment of dna from vibrio fischeri strain atcc7744 that contains all the information required to confer plasmid-borne, regulated bioluminescence upon strains of escherichia coli. the lux regulon from v. fischeri consists of two divergently transcribed operons, l (left) and r (right), and at least seven genes, luxr (l operon) and luxicdabe (r operon) and the intervening control region. the luxa and luxb genes encode respect ...19892801220
isolation of cdnas encoding gtp cyclohydrolase ii from arabidopsis thaliana.a gtp cyclohydrolase ii-encoding gene from arabidopsis thaliana was isolated through functional complementation of a mutant of escherichia coli, bsv18, deficient in this protein. the derived amino-acid sequence constitutes a polypeptide of 27 kda and shows 37-58% identity with previously published sequences of escherichia coli, bacillus subtilis, photobacterium leiognathi and p. phosphoreum.19957642114
formation of active bacterial luciferase between interspecific subunits in vivo.interspecific complementation between luxas and luxbs from vibrio harveyi, vibrio fischeri, photobacterium leiognathi and xenorhabdus luminescens was examined in vivo. the individual genes from these species were cloned on different compatible plasmids or amplified by pcr and brought together to yield cis combinations without extraneous dna. the beta subunits from v. harveyi and x. luminescens form active enzyme only with alpha subunits from one of these species. all other combinations yield act ...19957676858
13c and 15n nmr studies on the interaction between 6,7-dimethyl-8-ribityllumazine and lumazine protein.the interaction between the prosthetic group 6,7-dimethyl-8-(1'-d-ribityl)lumazine and the lumazine apoproteins from two marine bioluminescent bacteria, one from a relatively thermophilic species, photobacterium leiognathi, and the other from a psychrophilic species, photobacterium phosphoreum, was studied by 13c and 15n nmr using various selectively enriched derivatives. it is shown that the electron distribution in the protein-bound 6,7-dimethyl-8-ribityllumazine differs from that of free 6,7- ...19902331466
crystallographic characterization of a cu,zn superoxide dismutase from photobacterium leiognathi.crystals of a copper-zinc superoxide dismutase from photobacterium leiognathi, a luminescent marine bacterium that is the species-specific symbiont of the ponyfish, have been obtained from 2-methyl-2,4-pentanediol solutions. the space group was determined using screenless small-angle precession photographs, and was confirmed by analyzing area detector diffraction data with the xengen programs for indexing and refinement. the crystals are monoclinic, space group c2 (a = 126.4 a, b = 87.0 a, c = 4 ...19902325128
relationship of the luminous bacterial symbiont of the caribbean flashlight fish, kryptophanaron alfredi (family anomalopidae) to other luminous bacteria based on bacterial luciferase (luxa) genes.flashlight fishes (family anomalopidae) have light organs that contain luminous bacterial symbionts. although the symbionts have not yet been successfully cultured, the luciferase genes have been cloned directly from the light organ of the caribbean species, kryptophanaron alfredi. the goal of this project was to evaluate the relationship of the symbiont to free-living luminous bacteria by comparison of genes coding for bacterial luciferase (lux genes). hybridization of a lux ab probe from the k ...19902256783
a protein isolated from brucella abortus is a cu-zn superoxide dismutase.brucella abortus contains a protein that elicits an antigenic response in cattle previously exposed to the organism. the amino acid sequence of the recombinant form of this antigenic protein was determined by gas-phase sequencing of the pyridylethylated protein and its peptides obtained by digestion with cyanogen bromide (cnbr), clostripain, and staphylococcus aureus v8 protease. the brucella protein demonstrated 53.6% identity with the cu-zn superoxide dismutase (sod) from photobacterium leiogn ...19902105741
induction of superoxide dismutases in photobacterium leiognathi.we investigated the induction of cu,zn-sod (bacteriocuprein) and fe-sod in photobacterium leiognathi dk-a1 which was isolated from the light organ of the squid, droteuthis kensaki. the induction of superoxide dismutases depended on the addition of paraquat to the medium. induction of sod by paraquat was attributed mostly to the bacteriocuprein by measuring of the activities of both sods by using densitometry of isoelectrofocusing gel. when paraquat was added to the culture at various times in th ...19912071047
the rate of cu,zn superoxide dismutase evolution.the rate of amino acid replacement in cu,zn sod greatly departs from the expectations of the molecular clock. we examine 27 cu,zn sod sequences available and conclude that: (1) the sod enzymes from different mammal families differ from each other by roughly the same number of replacements, which is consistent with a simultaneous mammalian radiation; (2) over the most recent 60 million years (my) the rate of sod evolution is fairly high (15 aa/100 aa/100 myr) and may be considered constant; (3) t ...19912071040
growth and luminescence of luminous bacteria promoted by agents of microbial origin.the examination of four species of luminous bacteria photobacterium leiognathi, photobacterium phosphoreum, vibrio fischeri and vibrio harveyi has enabled us to reveal some nutrient medium components effecting growth, luminescence intensity and luciferase synthesis. these agents are nucleic components (nucleotides, nucleotides and amine bases), amino acids and vitamins, which are part of hydrolysates from the biomass of various lithotrophic microorganisms, hydrogen-oxidizing, iron-oxidizing and ...19938285107
evolutionary aspects of superoxide dismutase: the copper/zinc enzyme.copper/zinc superoxide dismutase is typically an enzyme of eukaryotes. the presence of the enzyme in the ponyfish symbiont photobacterium leiognathi and some free living bacteria does not have an immediate explanation. amino acid sequence alignment of 19 cu/zn superoxide dismutases shows 21 invariant residues in key positions related to maintenance of the beta-barrel fold, the active site structure including the electrostatic channel loop, and dimer contacts. nineteen other residues are invarian ...19912071039
the lumazine protein-encoding gene in photobacterium leiognathi is linked to the lux operon.the nucleotide (nt) sequence of the lump (embl accession no. x65612) gene of photobacterium leiognathi pl741 was determined and the amino acid (aa) sequence deduced. the encoded aa sequence of lump was identified as that of the lumazine protein (lump) by homology with that of photobacterium phosphoreum (56%). this small protein has a calculated m(r) of 19,997 and comprises 186 aa residues. biochemical studies suggested that lump is the protein which, when combined with luciferase, is responsible ...19938472956
sequence of the luxd gene encoding acyltransferase of the lux operon from photobacterium leiognathi.the nucleotide sequence of luxd (embl accession no. x65611), encoding acyltransferase (act), of the lux operon from photobacterium leiognathi pl741 was determined, and the amino acid (aa) sequence was deduced. act is a component of the fatty acid reductase complex, which is responsible for converting fatty acid to aldehyde that serves as the substrate in the luciferase-catalyzed bioluminescent reactions. the protein has a calculated m(r) of 34,384 and comprises 305 aa residues. alignment and com ...19938472957
the lux genes of the luminous bacterial symbiont, photobacterium leiognathi, of the ponyfish. nucleotide sequence, difference in gene organization, and high expression in mutant escherichia coli.the lux genes required for light expression in the luminescent bacterium photobacterium leiognathi (atcc 25521) have been cloned and expressed in escherichia coli and their organization and nucleotide sequence determined. transformation of a recombinant 9.5-kbp chromosomal dna fragment of p. leiognathi into an e. coli mutant (43r) gave luminescent colonies that were as bright as those of the parental strain. moreover, expression of the lux genes in the mutant e. coli was strong enough so that no ...19911915359
green flavoprotein from p. leiognathi: purification, characterization and identification as the product of the lux g(n) gene.a green flavoprotein (gfp) was isolated and purified to homogeneity from photobacterium leiognathi, strain 208. gfp is a homodimer of molecular weight 54,000 and contains two molecules of an unusual flavin per molecule of protein. various biochemical characteristics including isoelectric point, trypsin and chymotrypsin degradation, sds and temperature influence on subunit dissociation and the dissociation of the flavin chromophore, were investigated. the sequence of 23 n-terminal amino acids was ...19911746316
spectral properties and function of two lumazine proteins from photobacterium.the spectral properties are compared for two 6,7-dimethyl-8-ribityllumazine proteins from marine bioluminescent bacteria, one from a psychrophile, photobacterium phosphoreum, and the other from a thermophile, photobacterium leiognathi. the visible spectral properties, which are the ones by which the protein performs its biological function of bioluminescence emission, are almost the same for the two proteins: at 2 degrees c and 50 mm pi, ph 7, fluorescence quantum yield phi f = 0.59 and 0.54, re ...19853986186
direct measurement of excitation transfer in the protein complex of bacterial luciferase hydroxyflavin and the associated yellow fluorescence proteins from vibrio fischeri y1.time-resolved fluorescence was used to directly measure the energy transfer rate constant in the protein-protein complex involved in the yellow bioluminescence of vibrio fischeri, strain y1. in this reaction the putative donor is the fluorescent transient intermediate, luciferase hydroxyflavin, which exhibits a major fluorescence lifetime of the bound flavin of 10 ns. on addition of the acceptor, the v. fischeri yellow fluorescence protein containing either fmn or riboflavin as ligand, a rapid d ...19968679599
chemical characterization of lumazine protein from photobacterium leiognathi: comparison with lumazine protein from photobacterium phosphoreum.the properties of lumazine proteins purified from the marine bioluminescent bacteria photobacterium phosphoreum, a psychrophile, and photobacterium leiognathi, a relatively thermophilic species, are compared. an accurate 1:1 stoichiometry of binding of the ligand 6,7-dimethyl-8-ribityllumazine to each lumazine protein is established by back-titration of the apoprotein with the authentic ligand, using both fluorescence and absorption measurements. neither protein contains metal cofactors, organic ...19853986185
purification of lumazine proteins from photobacterium leiognathi and photobacterium phosphoreum: bioluminescence properties.bright strains of the marine bioluminescent bacterium photobacterium leiognathi produce a "lumazine protein" in amounts comparable to that previously found in photobacterium phosphoreum. new protocols are developed for the purification to homogeneity of the proteins from both species in yields up to 60%. in dimmer strains the amounts of lumazine protein in extracts are less, and also there is an accompanying shift of the bioluminescence spectral maximum to longer wavelength, 492 nm. both types o ...19853986184
interspecific luciferase beta subunit hybrids between vibrio harveyi, vibrio fischeri and photobacterium leiognathi.bacterial luciferase (ec 1.14.14.3) is a heterodimer composed of alpha- and beta-chains encoded by luxa and luxb, respectively. although some interspecific combinations of these subunits lead to active enzyme, others do not. the beta subunits of vibrio fischeri and photobacterium leiognathi form active enzyme with the alpha subunits of v.fischeri, p.leiognathi and vibrio harveyi, while the beta subunit from v.harveyi only complements the alpha subunit of v.harveyi. inactivity is caused by a lack ...19968888147
novel dimeric interface and electrostatic recognition in bacterial cu,zn superoxide dismutase.eukaryotic cu,zn superoxide dismutases (cuznsods) are antioxidant enzymes remarkable for their unusually stable beta-barrel fold and dimer assembly, diffusion-limited catalysis, and electrostatic guidance of their free radical substrate. point mutations of cuznsod cause the fatal human neurodegenerative disease amyotrophic lateral sclerosis. we determined and analyzed the first crystallographic structure (to our knowledge) for cuznsod from a prokaryote, photobacterium leiognathi, a luminescent s ...19968917495
the presence of a copper/zinc superoxide dismutase in the bacterium photobacterium leiognathi: a likely case of gene transfer from eukaryotes to prokaryotes.the free-living bacterium photobacterium leiognathi is also known to be a symbiont of ponyfish. the presence of a copper/zinc superoxide dismutase in p. leiognathi has been considered to be a case of gene transfer from eukaryotes to prokaryotes because this form of superoxide dismutase is normally present only in higher eukaryotic species. however, the amino acid sequence of the enzyme from the bacterium exhibited low identities (25-30%) with the same enzyme from eukaryotes. when amino acid muta ...19853855538
spectroscopic investigations of the single tryptophan residue and of riboflavin and 7-oxolumazine bound to lumazine apoprotein from photobacterium leiognathi.spectroscopic techniques have been applied to investigate the conformation, local structure, and dynamic properties of the apoprotein of the lumazine protein from photobacterium leiognathi and the holoprotein reconstituted with either the natural ligand 6,7-dimethyl-8-ribityllumazine or the closely related analogues riboflavin and 6-methyl-7-oxo-8-ribityllumazine (7-oxolumazine). the analogues are bound similarly to the natural prosthetic group. they exhibit similar shifts on binding in their ab ...19873828324
spectroscopic characterization of recombinant cu,zn superoxide dismutase from photobacterium leiognathi expressed in escherichia coli: evidence for a novel catalytic copper binding site.cu,zn superoxide dismutase from photobacterium leiognathi has been cloned and expressed in escherichia coli. the circular dichroism spectrum in the uv region of the recombinant protein indicates an higher content of random coil structure with respect to the eukaryotic enzymes. investigation of the active site by optical, cd, and epr spectroscopy indicates a different coordination geometry around the catalytic copper site with respect to the eukaryotic enzymes. in particular a different orientati ...19979188710
the pki gene encoding pyruvate kinase i links to the luxz gene which enhances bioluminescence of the lux operon from photobacterium leiognathi.partial 3'-end nucleotide sequence of the pki gene (genbank accession no. af019143) from photobacterium leiognathi atcc 25521 has been determined, and the encoded pyruvate kinase i is deduced. pyruvate kinase i is the key enzyme of glycolysis, which converts phosphoenol pyruvate to pyruvate. alignment and comparison of pyruvate kinase is from p. leiognathi, e. coli and salmonella typhimurium show that they are homologous. nucleotide sequence reveals that the pki gene is linked to the luxz gene t ...19979345300
analogs of the autoinducer of bioluminescence in vibrio fischeri.the enzymes for luminescence in vibrio fischeri are induced only when a sufficient concentration of a metabolic product (autoinducer) specifically produced by this species accumulates. it has previously been shown that the autoinducer is 3-oxohexanoyl homoserine lactone and that it enters the cells by simple diffusion. to further study the mechanism of induction, we have synthesized several analogs of the autoinducer. the analogs were tested with v. fischeri for their inducing activity and for t ...19863813773
physical characterization of lumazine proteins from photobacterium.the physicochemical properties of photobacterium lumazine proteins have been investigated. the molecular weights obtained by several physical techniques are in good agreement, and the averages are 2% and 8% higher than the minimum molecular weights from amino acid and ligand content. the average molecular weights, sedimentation coefficients, and molecular radii are respectively the following: photobacterium leiognathi lumazine protein, 21 200 +/- 300, 2.18 s, and 22.9 a; photobacterium phosphore ...19853986187
determination of rotational correlation times from deconvoluted fluorescence anisotropy decay curves. demonstration with 6,7-dimethyl-8-ribityllumazine and lumazine protein from photobacterium leiognathi as fluorescent indicators.the experimental and analytical protocols required for obtaining rotational correlation times of biological macromolecules from fluorescence anisotropy decay measurements are described. as an example, the lumazine protein from photobacterium leiognathi was used. this stable protein (mr 21 200) contains the noncovalently bound, natural fluorescent marker 6,7-dimethyl-8-ribityllumazine, which has in the bound state a long fluorescence lifetime (tau = 14 ns). shortening of the fluorescence lifetime ...19853986188
characteristic analysis of the luxg gene encoding the probable flavin reductase that resides in the lux operon of photobacterium leiognathi.nucleotide sequence of the luxg gene (genbank accession no. af053227) from photobacterium leiognathi pl741 has been determined, and the encoded probable flavin reductase is deduced. the probable flavin reductase encoded by the luxg gene has a calculated m(r) 26,544 and comprises 235 amino acid residues. the probable flavin reductase like the nad(p)h-flavin reductase might catalyze the reduction of flavins. alignment and comparison of the probable flavin reductases from p. leiognathi pl741, atcc ...19989610381
cu,zn superoxide dismutase from photobacterium leiognathi is an hyperefficient enzyme.the catalytic rate constant of recombinant photobacterium leiognathi cu,zn superoxide dismutase has been determined as a function of ph by pulse radiolysis. at ph 7 and low ionic strength (i = 0.02 m) the catalytic rate constant is 8.5 x 10(9) m-1 s-1, more than two times the value found for all the native eukaryotic cu,zn superoxide dismutases investigated to date. similarly, brownian dynamics simulations indicate an enzyme-substrate association rate more than two times higher than that found f ...19989724543
osmotic control of luminescence and growth in photobacterium leiognathi from ponyfish light organs.osmolarity was found to control the luminescence and growth of photobacterium leiognathi strain ln-1a isolated from the light organ of the ponyfish leiognathus nuchalis (family leiognathidae). low osmolarity (ca. 400 mosm) stimulated luminescence per cell 80 to 100-fold to a level (ca. 2.0 x 10(4) quanta . s-1 . cell-1) equal to that of bacteria taken directly from the light organ and increased the level of luciferase per cell 8 to 10-fold compared to high osmolarity (ca. 800 mosm). conversely, ...19853994483
bacteriocuprein superoxide dismutases in pseudomonads.two new instances of the rare bacteriocuprein form of superoxide dismutase have been discovered in pseudomonas diminuta and p. maltophilia. each species contains a manganese superoxide dismutase as well. eight other strains of pseudomonas and xanthomonas spp. lacked bacteriocupreins and contained either a manganese or an iron superoxide dismutase. native molecular weights and isoelectric points were determined for all these bacterial dismutases. a monospecific polyclonal antibody was prepared ag ...19853997777
toward the engineering of a super efficient enzyme.the catalytic activity of a mutant of photobacterium leiognathi cu, zn superoxide dismutase in which the glu59 residue, conserved in most bacterial variants of the enzyme, has been replaced by glutamine was investigated by pulse radiolysis. at neutral ph the enzyme was found to have a kcat/km of 1.0 +/- 0.1 x 10(10) m-1s-1 the highest value ever found for any superoxide dismutase. brownian dynamics simulation suggests that such a high value is due to an enhanced substrate attraction by the modif ...199910079201
comparative biochemistry of the cell envelopes of photobacterium leiognathi and escherichia coli.photobacterium leiognathi closely resembles escherichia coli with respect to cell lysis by lysozyme, and the fractionation of outer and cytoplasmic membranes. the two organisms differ in their phospholipid contents and, more significantly, in outer membrane protein compositions.19836352861
raman spectra of flavin bound in flavodoxins and in other flavoproteins. evidence for structural variations in the flavin-binding region.the resonance coherent anti-stokes raman scattering (cars) spectra for a number of flavoproteins are found to be fingerprints for the particular type of flavoprotein. one group studied were the bacterial flavodoxins: desulfovibrio vulgaris, desulfovibrio desulfuricans, azotobacter vinelandii, megasphaera elsdenii, clostridium kluyverii and clostridium formicoaceticum. the other examples were the enzymes lactate monooxygenase and glucose oxidase. fmn complexed to vibrio harveyi luciferase, and a ...19836840072
evidence of stable monomeric species in the unfolding of cu,zn superoxide dismutase from photobacterium leiognathi.the equilibrium unfolding process of photobacterium leiognathi cu,zn superoxide dismutase has been quantitatively monitored through circular dichroism (cd) and fluorescence spectroscopy, upon increasing the guanidinium hydrochloride concentration. the study has been undertaken for both the holo- and the copper-free derivative to work out the role of copper in protein stability. in both cases the unfolding was reversible. the denaturation curve derived from cd and fluorescence spectroscopy was no ...199910510278
a new, sensitive and simple bioluminescence test for mutagenic compounds.a spontaneous dark variant of the luminous bacterium photobacterium leiognathi was isolated. the reversion frequency of this variant to genetic-hereditary luminescent cells is greatly increased by nanogram quantities of different base-substitution and frameshift agents. this makes it possible to detect mutagenic compounds at concentrations 100 times lower than that detected by the ames test. curing agents, such as acridine dyes, ethidium bromide and sodium dodecyl sulfate, are also very active i ...19806990233
functional and crystallographic characterization of salmonella typhimurium cu,zn superoxide dismutase coded by the sodci virulence gene.the functional and three-dimensional structural features of cu,zn superoxide dismutase coded by the salmonella typhimurium sodci gene, have been characterized. measurements of the catalytic rate indicate that this enzyme is the most efficient superoxide dismutase analyzed so far, a feature that may be related to the exclusive association of the sodci gene with the most pathogenic salmonella serotypes. the enzyme active-site copper ion is highly accessible to external probes, as indicated by quen ...200010970746
single mutation at the intersubunit interface confers extra efficiency to cu,zn superoxide dismutase.the val28-->gly single mutant at the subunit interface of cu,zn superoxide dismutase from photobacterium leiognathi displays a k(cat)/k(m) value of 1.7x10(10) m(-1) s(-1), twice that of the native enzyme. analysis of the three-dimensional structure indicates that the active site cu,zn center is not perturbed, slight structural deviations being only localized in proximity of the mutation site. the enzyme-substrate association rate, calculated by brownian dynamics simulation, is identical for both ...200011033348
dna-damaging agents and dna-synthesis inhibitors induce luminescence in dark variants of luminous bacteria.the dna-damaging agents mitomycin c and uv irradiation, as well as the dna-synthesis inhibitors nalidixic acid, novobiocin and coumermycin, induce the de novo synthesis of luciferase and in vivo luminescence in dark variant cells of the luminous bacteria photobacterium leiognathi. mitomycin c and nalidixic acid also cause the induction of luminescence in wild-type cells in the absence of its natural inducer. in spite of the high level of in vivo luminescence of the treated dark-variant cells, no ...19817290100
the lumq gene is linked to the lump gene and the lux operon in photobacterium leiognathi.the nucleotide sequence of the designated lumq gene (embl accession no. u35231) from photobacterium leiognathi pl741 has been determined, and the encoded amino acid sequence is deduced. the lumq protein has a calculated m(r) of 28,416 and comprises 248 amino acid residues. the lumq gene is identified as the envy-like gene by significant similarity of the encoded protein with the envy and adiy proteins of e. coli; there the envy gene encodes the porin thermoregulatory protein envy, and the adiy g ...19957503752
common structural features of the luxf protein and the subunits of bacterial luciferase: evidence for a (beta alpha)8 fold in luciferase.the amino acid sequence identity and potential structural similarity between the subunits of bacterial luciferase and the recently determined structure of the luxf molecule are examined. the unique beta/alpha barrel fold found in luxf appears to be conserved in part in the luciferase subunits. from secondary structural predictions of both luciferase subunits, and from structural comparisons between the protein product of the luxf gene, nfp, and glycolate oxidase, we propose that it is feasible f ...19947703838
nucleotide sequence and functional analysis of regulatory region of the lump and the lux operon from photobacterium leiognathi.the lump gene is linked to the lux operon, but runs in the opposite direction in photobacterium leiognathi pl741. the gene order of the lump and the lux operon is < -lump-r & r-luxc-luxd-luxa-luxb-luxn-luxe- > (r & r: regulatory region). the nucleotide sequence of the regulatory region (827-bp) between the lump and the lux operon was determined. sequence analysis illustrates that the regulatory region includes two divergent promoter systems, pr-promoter system for the lux operon (r-operon) and p ...19957763266
structural refinement of the non-fluorescent flavoprotein from photobacterium leiognathi at 1.60 a resolution.the crystallographically-determined structure of the non-fluorescent flavoprotein (nfp) from photobacterium leiognathi, a homolog of the bacterial luciferase subunits, has been refined to a conventional r-factor [formula: see text] of 0.175 using synchrotron data between 10.0 and 1.60 a resolution. the molecular structure is a homodimer of beta/alpha domains, the monomer having structural similarities to (beta alpha)8 barrel proteins. however, one beta-strand and three alpha-helices of a typical ...19957776372
properties of recombinant fluorescent proteins from photobacterium leiognathi and their interaction with luciferase intermediates.ligand binding and luciferase interaction properties of the recombinant protein corresponding to the lumazine protein gene (embl x56534) of photobacterium leiognathi have been determined by fluorescence dynamics, circular dichroism, gel filtration, and sds-page. scatchard analysis of a fluorescence titration shows that the apoprotein possess one binding site, and at 30 degrees c the kds (microm) are as follows: 6,7-dimethyl-8-ribityllumazine, 0.26; riboflavin, 0.53; and much more weakly bound fm ...19957880825
expression and properties of the recombinant lumazine (riboflavin) protein from photobacterium leiognathi.photobacterium leiognathi lumazine protein has been expressed in escherichia coli in high yield, 30 mg/l. the cloned gene was one previously reported by illarionov (embl x56534), that had a similar sequence and was located in the same position as the lumazine protein gene in p. phosphoreum. this gene was placed downstream of the t7 gene 10 promoter of the plasmid pt7-7. when the e. coli are grown at 37 degrees c the protein accumulates in inclusion bodies but solubilization can be achieved in 6 ...19947947939
the lumazine synthase/riboflavin synthase complex of bacillus subtilis. x-ray structure analysis of hollow reconstituted beta-subunit capsids.the lumazine synthase/riboflavin synthase complex of bacillus subtilis consists of an icosahedral capsid of 60 beta subunits enclosing a triplet of alpha subunits. an x-ray structure of 0.32 nm resolution has been obtained for the icosahedral capsid of the native alpha 3 beta 60 complex [ladenstein, r., schneider, m., huber, r., bartunik, h. d., wilson, k., schott, k. & bacher, a. (1988) j. mol. biol. 203, 1045-1070]. beta subunits were isolated after denaturation of the alpha 3 beta 60 complex ...19948055941
nucleotide sequence of the luxc gene encoding fatty acid reductase of the lux operon from photobacterium leiognathi.the nucleotide sequence of the luxc gene (embl accession no. 65156) encoding fatty acid reductase (far) of the lux operon from photobacterium leiognathi pl741 was determined and the encoded amino acid sequence deduced. the fatty acid reductase is a component of the fatty acid reductase complex. the complex is responsible for converting fatty acid to aldehyde which serves as the substrate in the luciferase-catalyzed bioluminescent reaction. the protein comprises 478 amino acid residues and has a ...19938447834
crystal structure of a flavoprotein related to the subunits of bacterial luciferase.the molecular structure of the luxf protein from the bioluminescent bacterium photobacterium leiognathi has been determined by x-ray diffraction techniques and refined to a conventional r-factor of 17.8% at 2.3 a resolution. the 228 amino acid polypeptide exists as a symmetrical homodimer and 33% of the monomer's solvent-accessible surface area is buried upon dimerization. the monomer displays a novel fold that contains a central seven-stranded beta-barrel. the solvent-exposed surface of the mon ...19938491169
[bioluminescent analysis of the sos-response of escherichia coli cells].we constructed a recombinant plasmid ppls-1 to estimate the level of sos response in escherichia coli by the bioluminescent method. a 6.7-kb promoterless operon of bioluminescence from photobacterium leiognathi was cloned into a pbr322 vector, in which its expression was controlled by the sos promoter of gene cda from a plasmid cold. the sequence between the 5'-terminal sph1 site of the operon and start codon atg of the luxc gene was shown to be 56 bp in length and had no effect on the level of ...19968723628
interaction of photobacterium leiognathi and vibrio fischeri y1 luciferases with fluorescent (antenna) proteins: bioluminescence effects of the aliphatic additive.the kinetics of the bacterial bioluminescence reaction is altered in the presence of the fluorescent (antenna) proteins, lumazine protein (lump) from photobacterium or the yellow fluorescence proteins (yfp) having fmn or rf bound, from vibrio fischeri strain y1. depending on reaction conditions, the bioluminescence intensity and its decay rate may be either enhanced or strongly quenched in the presence of the fluorescent proteins. these effects can be simply explained on the basis of the same pr ...19968810914
dominance of vibrio fischeri in secreted mucus outside the light organ of euprymna scolopes: the first site of symbiont specificity.previous studies of the euprymna scolopes-vibrio fischeri symbiosis have demonstrated that, during colonization, the hatchling host secretes mucus in which gram-negative environmental bacteria amass in dense aggregations outside the sites of infection. in this study, experiments with green fluorescent protein-labeled symbiotic and nonsymbiotic species of gram-negative bacteria were used to characterize the behavior of cells in the aggregates. when hatchling animals were exposed to 10(3) to 10(6) ...200312839763
nucleotide sequence and functional analysis of the luxe gene encoding acyl-protein synthetase of the lux operon from photobacterium leiognathi.nucleotide sequence of the luxe gene genbank accession no. u66407 from photobacterium leiognathi pl741 has been determined, and the amino acid sequence of acyl-protein synthetase encoded by the luxe gene is deduced. nucleotide sequence reveals that the luxe gene encodes acyl-protein synthetase, which is a component of the fatty acid reductase complex that is responsible for converting fatty acid to aldehyde as substrate in the luciferase-catalyzed bioluminescence reaction. the acyl-protein synth ...19968941351
collaborative effects of photobacterium cuzn superoxide dismutase (sods) and human ap endonuclease in dna repair and sod-deficient escherichia coli under oxidative stress.the defenses against free radical damage include specialized repair enzymes that correct oxidative damage in dna and detoxification systems such as superoxide dismutases (sods). these defenses may be coordinated genetically as global responses. we hypothesized that the expression of sod and dna repair genes would inhibit dna damage under oxidative stress. therefore, protection of escherichia coli mutants deficient in sod and dna repair genes (sod-, xth-, and nfo-) was demonstrated by transformin ...200414744629
purification and characterization of flavoproteins and cytochromes from the yellow bioluminescence marine bacterium vibrio fischeri strain y1.several flavoproteins and cytochromes that occur as major components in extracts of the yellow bioluminescence y1 strain of the marine bacterium vibrio fischeri have been purified and characterized with respect to their mass (sds/page and matrix-assisted laser-desorption/ionization ms), chromatographic properties, n-terminal sequence, and spectroscopy (absorption, fluorescence emission and anisotropy decay). the investigated proteins were as follows: yellow fluorescence protein (yfp) with bound ...19979183020
phylogenetic analysis of the lux operon distinguishes two evolutionarily distinct clades of photobacterium leiognathi.the luminous marine bacterium photobacterium mandapamensis was synonymized several years ago with photobacterium leiognathi based on a high degree of phenotypic and genetic similarity. to test the possibility that p. leiognathi as now formulated, however, actually contains two distinct bacterial groups reflecting the earlier identification of p. mandapamensis and p. leiognathi as separate species, we compared p. leiognathi strains isolated from light-organ symbiosis with leiognathid fishes (i.e. ...200415034641
[effect of glucose on luciferase expression in photobacterium leiognathi].photobacterium leiognathi cultured in marine broth emits a luminescence that is temporarily enhanced and then extinguished by glucose. glucose reduces the luciferase level and the expression of lux abg mrna in p. leiognathi. the amount of atp in p. leiognathi is temporarily increased and then declines to the normal level. these results indicate that the extinguishing by glucose in p. leiognathi is induced by the interruption of the translation of luciferase.200415467278
experimental and simulative dissociation of dimeric cu,zn superoxide dismutase doubly mutated at the intersubunit surface.the equilibrium properties of dimeric photobacterium leiognathi cu,zn superoxide dismutase mutant bearing two negative charges in the amino acid clusters at the association interface has been studied, experimentally and computationally, and compared to those of the native enzyme. pressure-dependent dissociation is observed for the mutant, as observed by the fluorescence shift of the unique tryptophan residue located at the intersubunit surface. the spectral shift occurs slowly, reaching a platea ...200515681652
h-ns protein represses transcription of the lux systems of vibrio fischeri and other luminous bacteria cloned into escherichia coli.high expression in escherichia coli of the lux system cistron of a luminous bacteria under its own control has been accomplished only for the vibrio fischeri lux system at high cell density. mutation of the hns gene in e. coli has resulted in strong expression of the v. fischeri lux system at low cell density even in an rpos-deleted strain of e. coli that emits very low levels of luminescence. the e. coli double mutant, mc4110 hns::kan rpos::tet carrying the lux system of v. fischeri, developed ...19979353217
characterization of fe/mn-superoxide dismutase from diatom thallassiosira weissflogii: cloning, expression, and property.a cdna clone of 1114 bp encoding a putative mn-superoxide dismutase (mn-sod) from diatom thallassiosira weissflogii was cloned by the pcr technique. nucleotide sequence analysis of this cdna clone revealed that it was translated into 201 amino acid residues. when the sequence was compared with mn-sods from vibrio mimicus and escherichia coli, as well as two fe-sods from e. coli and photobacterium leiognathi, this sod showed higher homology to mn-sod. the amino acid residues required to coordinat ...200515740026
a biosensor for environmental genotoxin screening based on an sos lux assay in recombinant escherichia coli cells.a genetically controlled luminescent bacterial reporter assay, the sos lux test, was developed for rapid detection of environmental genotoxins. the bioassay is based on the recombinant plasmid ppls-1, which was constructed as a derivative of pbr322, carrying the promoterless luxcdabfe genes of photobacterium leiognathi downstream of a truncated cda gene from cold with a strong sos promoter. e. coli reca+ strains containing this construction are inducible to high levels of light production in the ...19979361425
immobilization as a technical possibility for long-term storage of bacterial biosensors.for applications in field experiments, the recombinant strain salmonella typhimurium ta1535 was immobilized to permit its immediate utilization after long storage periods. salmonella typhimurium ta1535 cells contain the plasmid that has an inducible sos promoter fused to a promoterless luxcdabfe operon from photobacterium leiognathi. the induction of bioluminescence occurs in the presence of the dna-damaging agent mitomycin c which stimulates the bacterial sos response. early stationary phase ce ...200515791471
characteristics analysis of the luza gene encoding chaperone from photobacterium leiognathi related to bioluminescence.nucleotide sequence of the luza gene (genbank accession no. af039303) from photobacterium leiognathi atcc 25521 (ncimb 2193) has been determined, and the chaperone encoded by the luza gene was deduced. the luza chaperone has a calculated m(r) 26,295 and comprises 230 amino acid residues; the hydrophobic alpha-helix n-terminal 21 amino acid residues mkktifallfmsvfi sypsfa is the leader peptide, therefore the matured luza chaperone has a calculated m(r) 23,871 and comprises 209 amino acid residues ...19989535753
phenotypic characterization of the marine pathogen photobacterium damselae subsp. piscicida.the taxonomic position of photobacterium damselae subsp. piscicida, the causative agent of fish pasteurellosis, is controversial as this organism has also been described as 'pasteurella piscicida'. to clarify the taxonomic position of the pathogen, a total of 113 p. damselae subsp. piscicida strains and 20 p. damselae subsp. damselae strains, isolated from different geographical areas and from the main affected fish species, were analysed using 129 morphological and biochemical tests, including ...19989828416
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