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comparison of the meta pathway operons on nah plasmid pww60-22 and tol plasmid pww53-4 and its evolutionary significance.the regulated meta pathway operon for the catabolism of salicylate on the naphthalene plasmid pww60-22 was cloned into the broad-host-range vector pkt230 on a 17.5 kbp bamhi fragment. the recombinant plasmid conferred the ability to grow on salicylate when mobilized into plasmid-free pseudomonas putida paw130. a detailed restriction map of the insert was derived and the locations of some of the genes were determined by subcloning and assaying for their gene products in escherichia coli and p. pu ...19883254935
in vitro and in vivo antibacterial activities of me1207, a new oral cephalosporin.me1207 (pivaloyloxymethyl ester of me1206) is a new oral cephalosporin. me1206 is (6r,7r)-7-[(z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)- acetamido]-3-[(z)-2-(4-methylthiazol-5-yl)-ethyl]-cephem-4-carboxy lic acid. the susceptibilities of about 1,600 clinical isolates to me1206 were determined by the agar dilution method. me1206 showed a broad spectrum of activity against gram-positive and gram-negative bacteria. me1206 was more active than cefaclor, t-2525, and cefixime against staphylococcus ...19883264132
comparison of the amino acid sequences of the transacylase components of branched chain oxoacid dehydrogenase of pseudomonas putida, and the pyruvate and 2-oxoglutarate dehydrogenases of escherichia coli.the nucleotide sequence of bkdb, the structural gene for e2b, the transacylase component of branched-chain-oxoacid dehydrogenase of pseudomonas putida has been determined and translated into its amino acid sequence. the start of bkdb was identified from the n-terminal sequence of e2b isolated from branched-chain-oxoacid dehydrogenase of the closely related species, p. aeruginosa. the reading frame was composed of 65.5% g + c with 82.3% of the codons ending in g or c. there was no intergenic spac ...19883046941
[genes encoding bacterial rna-polymerase. i. cloning of rpobc-operon of pseudomonas putida and its physical map].the p. putida rpobc operon, coding for beta and beta' subunits of rna polymerase, was cloned and its physical map constructed.19883048271
pseudomonas stutzeri ferredoxin: close similarity to azotobacter vinelandii and pseudomonas ovalis ferredoxins.the complete primary structure of pseudomonas stutzeri strain zobell ferredoxin was determined by a combination of protease digestion, edman degradation, and carboxypeptidase digestion and was: tfvvtdncikckytdcvevcpvdcfyegpnflvih pdecidcalcepecpaqaifsedevpedqqefielnadlaevwpnite kkdaladaeewdgvkdklqyler. the calculated molecular weight was 12,110 excluding iron and sulfur atoms. the amino acid sequence was highly homologous to those of azotobacter vinelandii and pseudomonas ovalis ferredoxins. it ...19883053681
purification and properties of carnitine dehydrogenase from pseudomonas putida.carnitine dehydrogenase (carnitine:nad+ oxidoreductase, ec 1.1.1.108) from pseudomonas putida ifp 206 catalyzes the oxidation of l-carnitine to 3-dehydrocarnitine. the enzyme was purified 72-fold to homogeneity as judged by polyacrylamide gel electrophoresis. the molecular mass of this enzyme is 62 kda and consists of two identical subunits. the isoelectric point was found to be 4.7. the carnitine dehydrogenase is specific for l-carnitine and nad+. the optimum ph for enzymatic activity in the ox ...19883058208
[cloning and expression of pseudomonas putida gene controlling the catechol-2,3-oxygenase activity in escherichia coli cells].the genes nahc and nahd from pseudomonas putida naphthalene degradation plasmid pbs286 were cloned on the vector puc19 in escherichia coli cells. the catechol-2,3-oxygenase activity observed in e. coli cells containing recombinant plasmid pbs955 demands the participation of 32 kd polypeptide which is apparently the product of the nahc gene. second polypeptide of molecular weight 34.5 kd is synthesized in pbs955 containing e. coli minicells and perhaps it is a nahd gene product. the data obtained ...19883058550
molecular cloning of 3-phenylcatechol dioxygenase involved in the catabolic pathway of chlorinated biphenyl from pseudomonas putida and its expression in escherichia coli.genes encoding 3-phenylcatechol dioxygenases were cloned from the chlorobiphenyl-degrading pseudomonas putida strain ou83, using broad-host-range cosmid vector pcp13. restriction enzyme analysis of dna from 2,3-dioxygenase-positive chimeric cosmids showed dna inserts ranging in size from 6.0 to 30 kilobases. the origin of the dna insert in hybrid clones was established by using 32p-labeled hybrid clones (poh101 and poh810). a 2.3-kilobase hindiii fragment was common to two clones. the 2,3-dioxyg ...19883063207
[nucleotide sequence of of rpob gene encoding the rna-polymerase beta-subunit in pseudomonas putida]. 19883069413
[nucleotide sequence of the rpoc-gene coding for rna polymerase beta'-subunit of pseudomonas putida]. 19883069416
cloning of genes for branched-chain keto acid dehydrogenase in pseudomonas putida. 19883071713
cytochrome p450: molecular architecture, mechanism, and prospects for rational inhibitor design.cytochromes p450 catalyze the insertion of one o2-derived oxygen atom into an aliphatic or aromatic molecule. p450s are best known for the metabolism of xenobiotic molecules, where hydroxylation renders insoluble hydrocarbons more soluble for easier elimination. in addition to this important catabolic function, p450s catalyze key steps in steroid and plant growth regulator metabolism. a variety of therapeutic, fungicidal, and agochemical agents that perturb these metabolic pathways very likely o ...19883073382
stability fluctuations of plasmid-bearing cells: immobilization effects.the maintenance of the plasmid vectors ptg201 and ptg206 (which both carry the pseudomonas putida xyle gene) and pb lambda h3 in escherichia coli hosts was studied in free and immobilized continuous cultures. ptg201, containing the strong lambda pr promoter, was more quickly lost than plasmid ptg206, containing the tetracycline resistance gene promoter. the instability of ptg201 seems to be related to high expression of the cloned xyle genet. fluctuations in the proportion of ptg201-containing c ...19883075659
cloning and expression of pca genes from pseudomonas putida in escherichia coli.beta-ketoadipate elicits expression of five structural pca genes encoding enzymes that catalyse consecutive reactions in the utilization of protocatechuate by pseudomonas putida. three derivatives of p. putida prs2000 were obtained, each carrying a single copy of tn5 dna inserted into a separate region of the genome and preventing expression of different sets of pca genes. selection of tn5 in or near the pca genes in these derivatives was used to clone four structural pca genes and to enable the ...19883076176
physical and functional mapping of two cointegrate plasmids derived from rp4 and tol plasmid pdk1.cointegrate plasmids were formed in vivo between the broad-host-range r-plasmid rp4 and two catabolic plasmids derived from pseudomonas putida hs1. one of these was the wild-type plasmid pdk1 encoding the complete inducible toluene/xylene (tol) catabolic pathway and one was pdkt1, a deletion derivative of pdk1 selected after growth of hs1 on benzoate and supporting growth on only toluene. the two plasmids formed, pdk2 and pdkt2 respectively, each consisted of a complete rp4 replicon in which was ...19883076182
[isolation and comparative characteristics of 2 unrelated temperate phages of pseudomonas putida ppg1].two temperate bacteriophages, pp56 and pp71, specific for bacteria of pseudomonas putida strain ppg1 have been isolated for the first time. characterization of the phages was performed. both of them accomplish stable lysogenization of p. putida ppg1 cells. the phages are inducible. several groups of clear plaque (c) mutants of pp56 and pp71 with altered processes of establishment and maintenance of lysogenic state have been isolated, according to complementation test. the phages differ in follow ...19883129337
purification and characterization of cutinase from a fluorescent pseudomonas putida bacterial strain isolated from phyllosphere.cutinase, an extracellular enzyme, was induced by cutin in a fluorescent pseudomonas putida strain that was found to be cohabiting with an apparently nitrogen-fixing corynebacterium. this enzyme was purified from the culture fluid by acetone precipitation followed by chromatography on deae-cellulose, qae-sephadex, sepharose 6b, and sephadex g-100. the purified enzyme showed a single band when subjected to polyacrylamide electrophoresis and the enzymatic activity coincided with the protein band. ...19883130804
physiological comparison of d-cysteine desulfhydrase of escherichia coli with 3-chloro-d-alanine dehydrochlorinase of pseudomonas putida cr 1-1.d-cysteine desulfhydrase of escherichia coli w3110 delta trped102/f' delta trped102 was physiologically characterized. it was found to be located in the cytosolic fraction, as 3-chloro-d-alanine dehydrochlorinase is. d-cysteine desulfhydrase catalyzed not only the alpha, beta-elimination reaction of o-acetyl-d-serine to form pyruvate, acetic acid and ammonia, but also the beta-replacement reaction of o-acetyl-d-serine with sulfide to form d-cysteine. however, these reactions appeared not to proc ...19883132906
transformation of bacteria with plasmid dna by electroporation.the possibility of electric field-mediated transformation ("electroporation") of a gram-positive bacterium (enterococcus faecalis) and two gram-negative bacteria (escherichia coli and pseudomonas putida) with plasmid dna was investigated. e. faecalis protoplasts could be transformed by electroporation with a transformation frequency of 10(4) to 10(5) transformants/micrograms plasmid. untreated--i.e., washed--cells of e. coli could be transformed with rates of 1 x 10(5) transformants/micrograms p ...19883133958
the complete amino acid sequence and identification of the active-site arginine peptide of escherichia coli 2-keto-4-hydroxyglutarate aldolase.the complete amino acid sequence of 2-keto-4-hydroxyglutarate aldolase from escherichia coli has been established in the following manner. after being reduced with dithiothreitol, the purified aldolase was alkylated with iodoacetamide and subsequently digested with trypsin. the resulting 19 peptide peaks observed by high performance liquid chromatography, which compared with 21 expected tryptic cleavage products, were all isolated, purified, and individually sequenced. overlap peptides were obta ...19883136164
[bacteriophages of pseudomonas putida containing single-stranded canonical dna breaks].it was shown that bacteriophage tf as well as bacteriophages phi p4/40, phi p25/42, phi p23/40 and phi p6/40, which are specific to different p. putida strains, contain the single strand breaks in their dna. the breaks are localized in one strand of dna molecules and are repairable with t4 dna ligase. bacteriophage tf has no detectable dna homology with phi p4/40, phi p25/42, phi p23/40 and phi p6/40 bacteriophages. all the phages studied have no relation with other known pseudomonas phages. bac ...19883137462
cloning and expression of the cata and catbc gene clusters from pseudomonas aeruginosa pao.a 9.9-kilobase (kb) bamhi restriction endonuclease fragment encoding the cata and catbc gene clusters was selected from a gene bank of the pseudomonas aeruginosa pao1c chromosome. the cata, catb, and catc genes encode enzymes that catalyze consecutive reactions in the catechol branch of the beta-ketoadipate pathway: cata, catechol-1,2-dioxygenase (ec 1.13.11.1); catb, muconate lactonizing enzyme (ec 5.5.1.1); and catc, muconolactone isomerase (ec 5.3.3.4). a recombinant plasmid, pro1783, which c ...19883139626
construction of a shuttle vector for inducible gene expression in escherichia coli and bacillus subtilis.the construction of a shuttle vector for inducible gene expression allowing fast and easy cloning in escherichia coli and subsequent transformation of bacillus subtilis is presented. the expression is based on the regulation of the tac promoter by the lac repressor which was assayed with the xyle gene from pseudomonas putida as a marker gene. the laciq gene, transcribed by the strong spo promoter, allowed full repression of the weak tac promoter.19883141570
survival of rifampin-resistant mutants of pseudomonas fluorescens and pseudomonas putida in soil systems.the fate of spontaneous chromosomal rifampin-resistant (rifr) mutants of pseudomonas putida and pseudomonas fluorescens in sterile and live organic soil from which they were isolated was studied. in sterile native-soil assays, a rifr mutant of p. putida showed no decrease in competitive fitness when compared with the wild-type parent. however, mutants of p. fluorescens were of two general categories. group 1 showed no difference from the wild type in terms of growth rate, competitive fitness, an ...19883144244
mineralization of the dibenzothiophene biodegradation products 3-hydroxy-2-formyl benzothiophene and dibenzothiophene sulfone.dibenzothiophene is degraded to 3-hydroxy-2-formyl benzothiophene by various bacteria, including a strain of pseudomonas putida that also forms dibenzothiophene sulfone via an alternate pathway. by using these end products as substrates, mixed enrichment cultures that could degrade 3-hydroxy-2-formyl benzothiophene and dibenzothiophene sulfone with the formation of co2 were established.19883146950
molecular studies on the role of a root surface agglutinin in adherence and colonization by pseudomonas putida.pseudomonas putida aggressively colonizes root surfaces and is agglutinated by a root surface glycoprotein. mutants of p. putida derived chemically or by tn5 insertion demonstrated enhanced or decreased agglutinability. two nonagglutinable tn5 mutants (agg) and two mutants with enhanced agglutinability (agg) possessed tn5 in unique restriction sites. agg mutants colonized root surfaces of seedlings grown from inoculated seeds, but at levels lower than those observed with the agg parent. in short ...198816347550
fusarium wilt suppression and agglutinability of pseudomonas putida.mutants of pseudomonas putida (agg) that lack the ability to agglutinate with components present in washes of bean and cucumber roots showed limited potential to protect cucumber plants against fusarium oxysporum f. sp. cucumerinum. however, a higher level of protection was observed against fusarium wilt in cucumber plants coinoculated with the parental bacterium (agg), which was agglutinable. the agg mutants did not colonize the roots of cucumber plants as extensively as the agg parental isolat ...198816347713
tn5-mediated cloning of a genetic region from pseudomonas putida involved in the stimulation of plant root elongation.transposon (tn5) mutagenesis was applied to pseudomonas putida gr12-2r3, which promotes root elongation (a phenotype designated pre) of brassica campestris under gnotobiotic conditions. of 3,000 tn5 transconjugants, only one mutant that lost pre activity but remained prototrophic and capable of plant root colonization was detected. this mutant was complemented by plasmid pre53, which contained a 15.0-kilobase dna insert isolated from a parental strain. the complemented mutant regained full pre a ...198816347807
thiosulfate oxidation by obligately heterotrophic bacteria.thiosulfate was oxidized stoichiometrically to tetrathionate during growth on glucose byklebsiella aerogenes, bacillus globigii, b. megaterium, pseudomonas putida, two strains each ofp. fluorescens andp. aeruginosa, and anaeromonas sp. a gram-negative, rod-shaped soil isolate, pseudomonad hw, converted thiosulfate to tetrathionate during growth on acetate. none of the organisms could use thiosulfate as sole energy source. the quantitative recovery of all the thiosulfate supplied to heterotrophic ...198824202996
effect of surface-active pseudomonas spp. on leaf wettability.different strains of pseudomonas putida and p. fluorescens isolated from the rhizosphere and phyllosphere were tested for surface activity in droplet cultures on polystyrene. droplets of 6 of the 12 wild types tested spread over the surface during incubation, and these strains were considered surface active; strains not showing this reaction were considered non-surface active. similar reactions were observed on pieces of wheat leaves. supernatants from centrifuged broth cultures behaved like dro ...198916347926
novel alterations in plasmid dna associated with aromatic hydrocarbon utilization by pseudomonas putida r5-3.subcultures of pseudomonas putida r5-3 altered their plasmid dna content in specific ways depending on the particular aromatic hydrocarbon utilized as the sole carbon source. two indigenous plasmids, 115 and 95 kilobases (kb) in size, were observed in r5-3a, which was derived from r5-3 by growth on minimal medium containing p-methylbenzoate as the sole carbon source. when r5-3a was transferred to medium containing m-xylene or toluene, derivative strains were obtained in which the 95-kb plasmid w ...198916347946
molybdenum involvement in aerobic degradation of 2-furoic acid by pseudomonas putida fu1.an organism identified as pseudomonas putida was isolated from an enrichment culture with 2-furoic acid as its sole source of carbon and energy. the organism contained a 2-furoyl-coenzyme a (coa) synthetase to form 2-furoyl-coa and a 2-furoyl-coa dehydrogenase to form 5-hydroxy-2-furoyl-coa as the first two enzymes involved in the degradation. tungstate, the specific antagonist of molybdate, decreased growth rate and consumption of 2-furoic acid but had no influence on growth with succinate. cor ...198916347977
degradation of acetonitrile by pseudomonas putida.a bacterium capable of utilizing high concentrations of acetonitrile as the sole source of carbon and nitrogen was isolated from soil and identified as pseudomonas putida. this bacterium could also utilize butyronitrile, glutaronitrile, isobutyronitrile, methacrylonitrile, propionitrile, succinonitrile, valeronitrile, and some of their corresponding amides, such as acetamide, butyramide, isobutyramide, methacrylamide, propionamide, and succinamide as growth substrates. acetonitrile-grown cells o ...198916348008
toxicity of trichloroethylene to pseudomonas putida f1 is mediated by toluene dioxygenase.trichloroethylene was metabolically activated by toluene dioxygenase to produce toxic effects in pseudomonas putida f1. cytotoxicity was indicated by growth inhibition and by the covalent modification of cellular molecules in p. putida f1 exposed to [c]trichloroethylene. with a toluene dioxygenase mutant, neither growth inhibition nor alkylation of intracellular molecules was observed.198916348039
response of plant-colonizing pseudomonads to hydrogen peroxide.colonization of plant root surfaces by pseudomonas putida may require mechanisms that protect this bacterium against superoxide anion and hydrogen peroxide produced by the root. catalase and superoxide dismutase may be important in this bacterial defense system. stationary-phase cells of p. putida were not killed by hydrogen peroxide (h(2)o(2)) at concentrations up to 10 mm, and extracts from these cells possessed three isozymic bands (a, b, and c) of catalase activity in native polyacrylamide g ...198916348058
antigenic crossreactivity between bacterial and plant cytochrome p-450 monoxygenases.although cytochrome p-450 monoxygenases mediate critical reactions in plant microsomes, characterization of their activities has been difficult due to their inherent instability and the lack of a crossreacting p-450 antibody. we have surveyed the effects of protein stabilizing agents on t-cinnamic acid hydroxylase (t-cah), a prominent microsomal p-450, and on total p-450 monoxygenase content. trans-cinnamic acid is the most effective protecting agent for t-cah activity. leupeptin, a broad spectr ...198916666804
phenoxyacetic acid degradation by the 2,4-dichlorophenoxyacetic acid (tfd) pathway of plasmid pjp4: mapping and characterization of the tfd regulatory gene, tfdr.plasmid pjp4 enables alcaligenes eutrophus jmp134 to degrade 3-chlorobenzoate and 2,4-dichlorophenoxyacetic acid (tfd). plasmid pro101 is a derivative of pjp4 obtained by insertion of tn1721 into a nonessential region of pjp4. plasmid pro101 was transferred by conjugation to several pseudomonas strains and to a. eutrophus aeo106, a cured isolate of jmp134. aeo106(pro101) and some pseudomonas transconjugants grew on tfd. transconjugants with a chromosomally encoded phenol hydroxylase also degrade ...19892914848
isolation of a third lipoamide dehydrogenase from pseudomonas putida.pseudomonads are the only organisms so far known to produce two lipoamide dehydrogenases (lpds), lpd-val and lpd-glc. lpd-val is the specific e3 component of branched-chain oxoacid dehydrogenase, and lpd-glc is the e3 component of 2-ketoglutarate and possibly pyruvate dehydrogenases and the l-factor of the glycine oxidation system. three mutants of pseudomonas putida, js348, js350, and js351, affected in lpdg, the gene encoding lpd-glc, have been isolated; all lacked 2-ketoglutarate dehydrogenas ...19892914869
demonstration, characterization, and mutational analysis of nahr protein binding to nah and sal promoters.the nahr gene of plasmid nah7 of pseudomonas putida encodes a 36-kilodalton polypeptide which activates transcription of the nah and sal operons in response to the inducer salicylate. a gel mobility shift assay was used to identify a dna-binding activity which was present only in extracts from either p. putida or escherichia coli containing a functional nahr gene. the binding activity was highly specific for dna containing the nah or sal promoters, but the apparent affinity for the promoters was ...19892914873
effect of degradative plasmid cam-oct on responses of pseudomonas bacteria to uv light.the effect of plasmid cam-oct on responses to uv irradiation was compared in pseudomonas aeruginosa, in pseudomonas putida, and in pseudomonas putida mutants carrying mutations in uv response genes. cam-oct substantially increased both survival and mutagenesis in the two species. p. aeruginosa strains without cam-oct exhibited much higher uv sensitivity than did p. putida strains. uv-induced mutagenesis of plasmid-free p. putida was easily detected in three different assays (two reversion assays ...19892914880
molecular cloning of salicylate hydroxylase genes from pseudomonas cepacia and pseudomonas putida.the sal gene encoding pseudomonas cepacia salicylate hydroxylase was cloned and the sal encoding pseudomonas putida salicylate hydroxylase was subcloned into plasmid vector pro2317 to generate recombinant plasmids ptk3 and ptk1, respectively. both cloned genes were expressed in the host pseudomonas aeruginosa pao1. the parental strain can utilize catechol, a product of the salicylate hydroxylase-catalyzed reaction, but not salicylate as the sole carbon source for growth due to a natural deficien ...19892916843
sequence analysis of the lpdv gene for lipoamide dehydrogenase of branched-chain-oxoacid dehydrogenase of pseudomonas putida.the production of two lipoamide dehydrogenases by pseudomonas is so far unique. one, lpd-val, is the specific e3 component of the branched-chain-oxoacid dehydrogenase and the second, lpd-glc, is the e3 component of 2-oxoglutarate dehydrogenase and the l-factor of the glycine oxidation system. the objective of the present research was to determine the nucleotide sequence of the structural gene for lpd-val in order to compare its deduced amino acid structure with that of other redox-active disulfi ...19892917566
crystal structure of muconolactone isomerase at 3.3 a resolution.the crystal structure of muconolactone isomerase from pseudomonas putida, a unique molecule with ten 96 amino acid subunits and 5-fold, and 2-fold symmetries, has been solved at 3.3 a resolution. the non-crystallographic symmetries were used to refine the initial single isomorphous replacement phases and produce an interpretable 10-fold averaged map. the backbone trace is complete and confirmed by the amino acid sequence fit. each subunit is composed of a body with two alpha-helices and an antip ...19892926818
analysis of nonpolar insertion mutations in the trfa gene of incp plasmid rk2 which affect its broad-host-range property.replication of broad-host-range plasmid rk2 requires the protein product(s) of the plasmid-encoded trfa gene to initiate replication at oriv, the vegetative replication origin. the trfa gene contains two translational starts which direct translation of two polypeptides, of 382 and 285 amino acids, which differ by the 97 amino acids at their n-terminus. nonpolar insertions which abolish expression of the larger trfa polypeptide but otherwise retain the trfa gene's normal expression signals severe ...19892495025
selection and characterization of a mutant of the cloned gene for mandelate racemase that confers resistance to an affinity label by greatly enhanced production of enzyme.the plasmid pscr1 containing the gene for mandelate racemase (ec 5.1.2.2) from pseudomonas putida (atcc 12633) allows pseudomonas aeruginosa (atcc 15692) to grow on (r)-mandelate as its sole carbon source [ransom, s. c., gerlt, j. a., powers, v. m., & kenyon, g. l. (1988) biochemistry 27, 540]; the chromosome of the p. aeruginosa host apparently does not contain the gene for mandelate racemase but does contain genes for the remaining enzymes in the mandelate pathway and enables growth on (s)-man ...19892496759
dna sequence of the tryptophan synthase genes of pseudomonas putida.genes encoding the 2 subunits of tryptophan synthase in pseudomonas putida have been identified and cloned by their similarity to the corresponding genes in pseudomonas aeruginosa. the deduced amino acid sequences were confirmed by comparison with regions ascertained earlier by protein sequencing. the pseudomonas amino acid sequences are 85% identical for the beta subunit and 70% identical for the alpha subunit. these sequences are compared to those of salmonella typhimurium, where the structure ...19892503057
plasmid control of the pseudomonas aeruginosa and pseudomonas putida phenotypes and of linalool and p-cymene oxidation.two pseudomonas strains (ppg777 and pag158) were derived from the parent isolate pseudomonas incognita (putida). strain ppg777 resembles the parental culture in growth on linalool as a source of carbon and slight growth on p-cymene, whereas pag158 grows well on p-cymene, but not on linalool or other terpenes tested, and has a p. aeruginosa phenotype. curing studies indicate that linalool metabolism is controlled by an extrachromosomal element whose loss forms a stable strain pag158 with the p-cy ...19892504698
synthesis of poly-3-hydroxyalkanoates is a common feature of fluorescent pseudomonads.the fluorescent pseudomonads are classified as a group, one characteristic of which is that they do not accumulate poly-3-hydroxybutyrate (phb) during nutrient starvation in the presence of excess carbon source. in this paper we show that prototype strains from this subclass, such as pseudomonas aeruginosa, pseudomonas putida, and pseudomonas fluorescens, do accumulate poly-3-hydroxyalkanoates (pha) when grown on fatty acids. these phas are composed of medium-chain-length (c6 to c12) 3-hydroxy f ...19892506811
microbial metabolism of quinoline and related compounds. ii. degradation of quinoline by pseudomonas fluorescens 3, pseudomonas putida 86 and rhodococcus spec. b1.quinoline catabolism was investigated with different bacterial strains, able to use quinoline as sole source of carbon, nitrogen and energy. some degradation products of quinoline were isolated from the culture fluids and identified. with pseudomonas fluorescens and pseudomonas putida we found 2-oxo-1,2-dihydroquinoline, 8-hydroxy-2-oxo-1,2-dihydroquinoline, 8-hydroxycoumarin and 2,3-dihydroxyphenylpropionic acid as intermediates. with a rhodococcus strain 2-oxo-1,2-dihydroquinoline, 6-hydroxy-2 ...19892514722
factors affecting conjugal transfer of plasmids encoding mercury resistance from pure cultures and mixed natural suspensions of epilithic bacteria.sixty-five pure cultures of epilithic bacteria were examined for their ability to transfer mercury resistance to pseudomonas aeruginosa; five isolates transferred plasmids encoding mercury resistance with frequencies ranging from 8.4 x 10(-8) to 2.8 x 10(-3) per recipient. two of the plasmids, pqm3 and pqm4, encoded narrow-spectrum mercury resistance, pqm3 also encoded streptomycin resistance, and both plasmids were broad host range. maximum transfer frequencies of epilithic plasmids from pure c ...19892515247
[expression of the phospholipase c gene of pseudomonas aeruginosa in escherichia coli and pseudomonas].the plc gene for phospholipase of pseudomonas aeruginosa, able to be transcribed only from its own promoter, has been introduced into escherichia coli, pseudomonas aeruginosa and pseudomonas putida cells in the recombinant plasmid ppms21 of a wide host range. the expression of plc gene in all recipient cells has been shown to be phosphate regulated. the fact emphasizes the identity of pho-regulation systems in escherichia coli and pseudomonas cells. the level of phospholipase activity is similar ...19892515430
selective medium for pseudomonas aeruginosa that uses 1,10-phenanthroline as the selective agent.the mic of 1,10-phenanthroline for 35 pseudomonas aeruginosa strains was 128 micrograms/ml, whereas 32 micrograms or less per ml inhibited all other microorganisms tested. on the basis of these results, a selective agar for p. aeruginosa which contained 15 g of trypticase soy broth (bbl microbiology systems), 15 g of agar, and 0.1 g of phenanthroline per liter was formulated. forty-four p. aeruginosa strains yielded a mean efficiency of plating on this medium of 79% of the counts obtained on try ...19892515804
growth of genetically engineered pseudomonas aeruginosa and pseudomonas putida in soil and rhizosphere.the effect of the addition of a recombinant plasmid containing the pgla gene encoding an alpha-1,4-endopolygalacturonase from pseudomonas solanacearum on the growth of pseudomonas aeruginosa and pseudomonas putida in soil and rhizosphere was determined. despite a high level of polygalacturonase production by genetically engineered p. putida and p. aeruginosa, the results suggest that polygalacturonase production had little effect on the growth of these strains in soil or rhizosphere.19892515805
cloning and analysis of genes involved in coenzyme b12 biosynthesis in pseudomonas denitrificans.cobalamin synthesis probably requires 20 to 30 different enzymatic steps. pseudomonas putida and agrobacterium tumefaciens mutants deficient in cobalamin synthesis (cob have been isolated. in p. putida, cob mutants were identified as being unable to use ethanolamine as a source of nitrogen in the absence of added cobalamin (deamination of ethanolamine requires coenzyme b12 as a cofactor). in a. tumefaciens, cob mutants were simply screened for their reduced cobalamin synthesis. a genomic library ...19892536665
cloning and characterization of a gene encoding an outer membrane protein required for siderophore-mediated uptake of fe3+ in pseudomonas putida wcs358.in iron-limited environments plant-growth-stimulating pseudomonas putida wcs358 produces a yellow-green fluorescent siderophore called pseudobactin 358. ferric pseudobactin 358 is efficiently taken up by cells of wcs358 but not by cells of another rhizophere-colonizing strain, pseudomonas fluorescens wcs374. a gene bank containing partial sau3a dna fragments from wcs358 was constructed in a derivative of the broad-host-range cosmid plafr1. by mobilization of this gene bank to strain wcs374 a cos ...19892540157
structure of the dnaa region of pseudomonas putida: conservation among three bacteria, bacillus subtilis, escherichia coli and p. putida.we have cloned from pseudomonas putida a gene homologous to escherichia coli dnaa, and determined the sequence of the gene and its neighboring region. the dnaa gene and at least three other genes, dnan, recf and gyrb, were found to be highly homologous to the genes in the dnaa regions of the e. coli and bacillus subtilis chromosomes. a non-translatable region of some 600 bp immediately upstream of the dnaa gene is also conserved in the three bacteria and contains 3, 12, and 14 dnaa-boxes (ttatcc ...19892540413
omegon-km: a transposable element designed for in vivo insertional mutagenesis and cloning of genes in gram-negative bacteria.to combine the features of the omega interposons with the advantages of in vivo transposition mutagenesis, we have constructed an artificial transposon, called omegon-km. the omegon-km transposon is carried on the plasmid pjff350 which can be conjugally mobilized into a broad range of gram-negative bacteria. omegon-km is flanked, in inverted orientation, by synthetic 28-bp repeats derived from the ends of is1. in addition, each end of omegon-km has the very efficient transcription and translatio ...19892546859
purification and some properties of a 2fe ferredoxin in pseudomonas ovalis.a [2fe-2s] ferredoxin was found in pseudomonas ovalis which was grown in a medium supplemented with glucose and ammonium sulfate. the molecular weight of the 2fe ferredoxin was estimated to be 13,000. it contained 2.2 gramatoms of non-heme iron and 2.3 gramatoms of acid-labile sulfur per mole protein. the absorption and circular dichroism spectra were characteristic of those of [2fe-2s] type ferredoxins, especially adrenodoxin and putidaredoxin. the electron paramagnetic resonance spectrum of th ...19892548508
a simple procedure for transferring genes cloned in escherichia coli vectors into other gram-negative bacteria: phenotypic analysis and mapping of tol plasmid gene xylk.a simple method to transfer non-conjugative escherichia coli plasmids to other gram-negative bacteria and their maintenance is described. this method involves generation of inverse transposition-mediated cointegrates of the non-conjugative e. coli plasmid with a conjugative incw broad-host-range plasmid, r388, carrying tn10. isolation of such cointegrates was readily effected by conjugal transfer from an e. coli donor containing the two plasmids to an e. coli recipient, with selection for transc ...19892548929
characterization of five genes in the upper-pathway operon of tol plasmid pww0 from pseudomonas putida and identification of the gene products.the upper operon of the tol plasmid pww0 of pseudomonas putida encodes a set of enzymes which transform toluene and xylenes to benzoate and toluates. the genetic organization of the operon was characterized by cloning of the upper operon genes into an expression vector and identification of their products in escherichia coli maxicells. this analysis showed that the upper operon contains at least five genes in the order of xylc-xylm-xyla-xylb-xyln. between the promoter of the operon and xylc, the ...19892549010
cloning of pmol28-encoded nickel resistance genes and expression of the genes in alcaligenes eutrophus and pseudomonas spp.the 163-kilobase-pair (kb) plasmid pmol28, which determines inducible resistance to nickel, cobalt, chromate, and mercury salts in its native host alcaligenes eutrophus ch34, was transferred to a derivative of a. eutrophus h16 and subjected to cloning procedures. after tn5 transposon mutagenesis, restriction endonuclease analysis, and dna-dna hybridization, two dna fragments, a 9.5-kb kpni fragment and a 13.5-kb hindiii fragment (hki), were isolated. hki contained ek1, the kpni fragment, as a su ...19892549012
the purification and characterization of 4-ethylphenol methylenehydroxylase, a flavocytochrome from pseudomonas putida jd1.the enzyme 4-ethylphenol methylenehydroxylase was purified from pseudomonas putida jd1 grown on 4-ethylphenol. it is a flavocytochrome c for which the mr was found to be 120,000 by ultracentrifuging and 126,000 by gel filtration. the enzyme consists of two flavoprotein subunits each of mr 50,000 and two cytochrome c subunits each of mr 10,000. the redox potential of the cytochrome is 240 mv. hydroxylation proceeds by dehydrogenation and hydration to give 1-(4'-hydroxyphenyl)ethanol, which is als ...19892556994
transposable elements for efficient manipulation of a wide range of gram-negative bacteria: promoter probes and vectors for foreign genes.we describe here the construction and use of a series of modified transposons based on the insertion sequence is1. like their parent, omegon-km [fellay et al., gene 76 (1989) 215-226], these elements permit efficient insertional mutagenesis of a variety of gram-negative bacteria. the presence of a functional pbr322 origin of replication within the transposable element facilitates subsequent cloning of the mutated gene. the omegon-km system was previously shown to function in pseudomonas putida, ...19892559879
[cloning and gene expression determining phenol breakdown in pseudomonas putida strains]. 19892561419
study of the 5-oxoprolinase reaction by 13c nmr.5-oxoprolinase catalyzes the atp-dependent decyclization of 5-oxo-l-proline to l-glutamate. previous studies provided evidence for the intermediate formation of a phosphorylated form of 5-oxoproline (seddon, a. p., and meister, a. (1986) j. biol. chem. 261, 11538-11541) and of a tetrahedral intermediate (li, l., seddon, a. p., and meister, a. (1987) j. biol. chem. 262, 11020-11025). a new approach to the study of the reaction mechanism using the 18o isotope effect on the 13c nmr signals for 5-ox ...19892563377
thermal activation of photoactivatable urocanase from pseudomonas putida.the dark inactivation of urocanase from pseudomonas putida is caused by the formation of a sulfite adduct of the tightly bound coenzyme, nicotinamide adenine dinucleotide. photodissociation of this adduct by uv radiation restores the enzyme activity. based on cold exhaustive dialysis the modification reaction appeared to be irreversible. however, we now report that sulfite modification of urocanase is reversible at higher temperatures. an arrhenius plot of the thermal activation is linear (20-38 ...19892570140
the stoichiometry of the tightly bound nad+ in urocanase. separation and characterization of fully active and inhibited forms of the enzyme.1. urocanase, purified by classical methods [keul, v., kaeppeli, f., ghosh, c., krebs, t., robinson, j. a. and rétey, j. (1979) j. biol. chem. 254, 843-851] from pseudomonas putida was submitted to high-performance liquid chromatography on a tsk-deae column. the enzyme was eluted in three resolved peaks (a, b and c) exhibiting specific activities of 3.4 u/mg, 1.85 u/mg and 0.4 u/mg, respectively. 2. the difference spectra of peaks b and a as well as of c and a showed maxima at 330 nm. 3. irradia ...19892574107
specific inhibition of bacterial and bovine urocanases by glycylglycine.urocanase (ec 4.2.1.49) purified from pseudomonas putida was unexpectedly inhibited by the dipeptide glycylglycine. using a spectrophotometric assay for urocanase activity, we characterized the inhibition. the inhibition was temperature-, concentration-, and time-dependent; 0.1, 0.5 and 1.0 mm glycylglycine inhibited the enzyme by 20%, 50% and 78%, respectively, in 60 min at 30 degrees c. dithiothreitol and reduced glutathione did not prevent the process. the inhibition was a pseudo first-order ...19892577699
operon structure and nucleotide homology of the chlorocatechol oxidation genes of plasmids pjp4 and pac27.alcaligenes eutrophus harboring plasmid pjp4 (strain jmp134) is capable of growing on both 2,4-dichlorophenoxyacetate (2,4-d) and 3-chlorobenzoate (3-cba), while pseudomonas putida carrying plasmid pac27 (strain ac867) can utilize only 3-cba as the sole carbon source. the tfdcdef operon of the pjp4 plasmid and the clcabd operon of plasmid pac27 each encode enzymes for the degradation of chlorocatechols (clc), key intermediates in the catabolism of 2,4-d and 3-cba. similarities in the nucleotide ...19892583528
survival of and plasmid stability in pseudomonas and klebsiella spp. introduced into agricultural drainage water.cell survival and plasmid stability in pseudomonas fluorescens r2f and pseudomonas putida cym 318 containing respectively, plasmid rp4 and prk2501, and klebsiella aerogenes nctc 418 harboring plasmid pbr322 were studied in sterile and nonsterile agricultural drainage water under both aerobic and anaerobic conditions and in the absence and presence of added nutrients. both pseudomonas strains survived well in sterile drainage water incubated aerobically, with or without added nutrients. however, ...19892590305
xyle functions as an efficient reporter gene in streptomyces spp.: use for the study of galp1, a catabolite-controlled promoter.we describe the development of a convenient and sensitive reporter gene system for streptomyces spp. based on the use of a promoterless copy of the xyle gene of pseudomonas putida. the xyle gene product is a catechol dioxygenase, which converts the colorless substrate catechol to an intensely yellow hydroxymuconic semialdehyde. a promoterless copy of xyle was placed under the transcriptional control of galp1, a glucose-repressed and galactose-induced promoter from streptomyces lividans, and its ...19892592344
molecular cloning, coding nucleotides and the deduced amino acid sequence of p-450bm-1 from bacillus megaterium.the gene encoding barbiturate-inducible cytochrome p-450bm-1 from bacillus megaterium atcc 14581 has been cloned and sequenced. an open reading frame in the 1.9 kb of cloned dna correctly predicted the nh2-terminal sequence of p-450bm-1 previously determined by protein sequencing, and, in toto, predicted a polypeptide of 410 amino acid residues with an mr of 47,439. the sequence is most, but less than 27%, similar to that of p-450cam from pseudomonas putida, so that p-450bm-1 clearly belongs to ...19892597681
[study of morphology and genome structure of pseudomonas putida bacteriophages for their classification].a group of 27 bacteriophages specific for pseudomonas putida strains ppg1 and ppn has been isolated. the phages were characterized and compared with the previously described virulent (pf 16, af, tf and pmw) and temperate (pp56 and pp71) phages. the new phages belong to b1 and c1 morphotypes, according to ackerman's classification. phage dnas were digested with several endonucleases; the molecular weights and homology of the dnas were determined. all phages of p. putida isolated up to now were di ...19892599372
nucleotide and deduced amino acid sequence of the rpon sigma-factor of pseudomonas putida. 19892602128
survival of pseudomonas putida uwc1 containing cloned catabolic genes in a model activated-sludge unit.the possibility of the accidental or deliberate release of genetically engineered microorganisms into the environment has accentuated the need to study their survival in, and effect on, natural habitats. in this study, pseudomonas putida uwc1 harboring a non-self-transmissible plasmid, pd10, encoding the breakdown of 3-chlorobenzoate was shown to survive in a fully functioning laboratory-scale activated-sludge unit (asu) for more than 8 weeks. the asu maintained a healthy, diverse protozoal popu ...19892604401
monohydroxylation of phenol and 2,5-dichlorophenol by toluene dioxygenase in pseudomonas putida f1.pseudomonas putida f1 contains a multicomponent enzyme system, toluene dioxygenase, that converts toluene and a variety of substituted benzenes to cis-dihydrodiols by the addition of one molecule of molecular oxygen. toluene-grown cells of p. putida f1 also catalyze the monohydroxylation of phenols to the corresponding catechols by an unknown mechanism. respirometric studies with washed cells revealed similar enzyme induction patterns in cells grown on toluene or phenol. induction of toluene dio ...19892604403
cloning and nucleotide sequences of nadh-putidaredoxin reductase gene (cama) and putidaredoxin gene (camb) involved in cytochrome p-450cam hydroxylase of pseudomonas putida.pseudomonas putida ppgl, which carries the cam plasmid encoding enzymes involved in the degradation pathway of d-camphor, can utilize d-camphor as a sole carbon source. cytochrome p-450cam and related enzymes participate in the early oxidation steps of d-camphor degradation metabolism. we cloned from a hindiii dna library of ppgl a 2.9 kbp cam segment which carries the major part of cama gene encoding nadh-putidaredoxin reductase and the entire camb gene encoding putidaredoxin. the 2.9 kbp cam s ...19892613690
[plasmids for biphenyl, chlorobiphenyl and metatoluylate degradation from pseudomonas putida].pseudomonas putida strain su83, harbors the pbs311 plasmid coding for the degradation of biphenyl, 2- and 4-chlorbiphenyl, meta- and paratoluylate. the insertional mutants of the plasmid obtained by the transposon tn5 insertion were isolated. one of the mutants was used for cloning of the biphenyl degradation genes. the plasmid pbs311:: tn5 dna was inserted into the bamhi site of the plasmid pbr322 and cloned. 11 recombinants of 354 tested were treated with 0.1% solution of 2,3-dioxybiphenyl. on ...19892628753
[genetic determination of degradation of ampholytic surfactants].plasmid dna was detected in pseudomonas putida 141 and p. stutzeri at strains which caused destruction of the ampholytic surfactants alkylamino-bis-propionate (aabp) and amidobetaine, respectively. as was demonstrated using genetic analytic procedures, the plasmids controlled aabp and amidobetaine destruction. no plasmid dna was found in p. desmolytica c37 which caused cyclimide destruction or in pseudomonas sp. 1 and citrobacter freundii to strains responsible for aabp destruction. apparently, ...19892636974
extracellular product of nocardia amarae induces bacterial cell flocculation.the fact that nocardia amarae yk1 produced a bacterial flocculation-inducing substance (designated as fix) was discovered. fix had a function of flocculating proliferous cells. fix-induced flocculation was inhibited by making cells resting, but not completely by adding chloramphenicol. fix worked widely on gram-positive to -negative bacteria. in the presence of fix, achromobacter cycloclastus iam1013, acinetobacter calcoaceticus iam1517, bacillus subtilis iam1069, escherichia coli c600-1, e. col ...19892653957
in vivo enzymology: a deuterium nmr study of formaldehyde dismutase in pseudomonas putida f61a and staphylococcus aureus.high-resolution deuterium nmr spectroscopy has been used to follow the detoxifying metabolism of [d2]formaldehyde in vivo in several bacterial species. production of [d2]methanol in escherichia coli confirms that the oxidation and reduction pathways of metabolism are independent in this organism. efficient production of equimolar quantities of [d]formate and [d3]methanol in pseudomonas putida f61a and staphylococcus aureus implicates a formaldehyde dismutase, or "cannizzarase", activity. these o ...19892655705
cloning and expression in escherichia coli of the toluene dioxygenase gene from pseudomonas putida ncib11767.the genes encoding toluene dioxygenase, toluene cis-glycol dehydrogenase and catechol 2.3-oxygenase from pseudomonas putida ncib 11767 were cloned and expressed in escherichia coli hb101 on a 20 kb fragment. the recombinant strain produced indigo and a variety of other coloured products. although the enzymes were expressed in the absence of inducers, further induction was observed in the presence of toluene or benzene, implying the presence of regulatory elements on the 20 kb insert.19892656389
5-carboxymethyl-2-hydroxymuconic semialdehyde dehydrogenases of escherichia coli c and klebsiella pneumoniae m5a1 show very high n-terminal sequence homology.5-carboxymethyl-2-hydroxymuconic semialdehyde (chms) dehydrogenase from escherichia coli c and klebsiella pneumoniae m5a1 have been purified and some of their properties studied. the apparent km values for nad and chms were 11.7 +/- 1.5 microm and 5.2 +/- 1.9 microm, respectively, for the k. pneumoniae enzyme, and 19.5 +/- 2.7 microm and 9.2 +/- 1.4 microm, respectively, for the e. coli enzyme. both enzymes were optimally active at ph 7.5 in sodium phosphate buffer. they had subunit molecular we ...19892656390
[cloning of pseudomonas putida genes responsible for the primary stages of oxidation of naphthalene in escherichia coli cells].data on cloning pseudomonas putida d-plasmid pbs286 (incp-9) genes which are responsible for primary stages of naphthalene oxidation as well as data on the expression of these genes in escherichia coli cells are presented. recombinant plasmid pbs959 containing the whole constitutive naha locus encoding naphthalene dioxygenase, a key enzyme of the naphthalene oxidation pathway, has been constructed on the basis of the puc19 vector. an evidence has been obtained that at least a portion of the sequ ...19892661326
cloning of a carbofuran hydrolase gene from achromobacter sp. strain wm111 and its expression in gram-negative bacteria.a 14-kilobase-pair (kbp) ecori dna fragment that encodes an enzyme capable of rapid hydrolysis of n-methylcarbamate insecticides (carbofuran hydrolase) was cloned from carbofuran-degrading achromobacter sp. strain wm111. when used to probe southern blots containing plasmid and total dnas from wm111, this 14-kbp fragment hybridized strongly to a 14-kbp ecori fragment from the greater than 100-kbp plasmid harbored by this strain but weakly to ecori-digested total dna from achromobacter sp. strain ...19892661544
2-oxoaldehyde metabolism in microorganisms.the properties of methylglyoxal-metabolizing enzymes in prokaryotic and eukaryotic microorganisms were studied systematically and compared with those of mammalian enzymes. the enzymes constitute a glycolytic bypass and convert methylglyoxal into pyruvate via lactate. the first step in this conversion is catalyzed by glyoxalase i, methylglyoxal reductase, or methylglyoxal dehydrogenase. the regulation of the yeast glyoxalase system was analyzed. the system was closely related to the proliferative ...19892663129
[susceptibilities of clinical isolates to antibacterial agents. a study mainly focused on ofloxacin (the second report). reported by the research group for testing ofloxacin susceptibility on clinical isolates].susceptibilities of various clinical isolates to ofloxacin (oflx) and other antibacterial drugs were examined at 128 hospital laboratories in 36 prefectures throughout japan between april, 1986 and march, 1987. the results were totalized with an emphasis mainly on oflx and were compared with data obtained in the previous year. in this study, identification and susceptibility tests of the isolates were carried out at each hospital laboratory and the tests were performed according to the 1-dilutio ...19892664255
identification of multiple repressor recognition sites in the hut system of pseudomonas putida.the hutc gene in pseudomonas putida encodes a repressor protein that negatively regulates the expression of all hut genes. we have overexpressed this cloned hutc gene in escherichia coli to identify p. putida hut regions that could specifically bind the repressor. ten restriction fragments, some of which were partially overlapping and spanned the coding portions of the p. putida hut region, were labeled and tested for their ability to recognize repressor in a filter binding assay. this procedure ...19892666390
involvement of pseudomonas putida rpon sigma factor in regulation of various metabolic functions.the rpon protein was originally identified in escherichia coli as a sigma (sigma) factor essential for the expression of nitrogen regulons. in the present study we cloned the pseudomonas putida rpon gene and identified its gene product as a protein with an apparent molecular weight of 78,000. a mutant rpon gene was constructed by in vitro insertion mutagenesis with a kanamycin cassette. a p. putida rpon mutant was then isolated by replacement of the intact chromosomal rpon gene by the mutant rpo ...19892666396
differences between the manganese- and the iron-containing superoxide dismutases of escherichia coli detected through sedimentation equilibrium, hydrodynamic, and spectroscopic studies.the genome of escherichia coli codes for two superoxide dismutases that may contain either iron (fesod) or manganese (mnsod) at the active site. the crystal structures of mnsods from two bacterial sources (but not e. coli) have been completed, and structural comparisons with the crystal structure of the fesod from either e. coli or pseudomonas ovalis have been made. despite the low degree (less than 50%) of sequence homology between the e. coli enzymes, the two proteins are suggested to be struc ...19892669953
transcription initiation at multiple promoters of the pfl gene by e sigma 70-dependent transcription in vitro and heterologous expression in pseudomonas putida in vivo.in vitro transcription experiments were used to provide further evidence that the gene encoding pyruvate formate-lyase (ec 2.3.1.54) from escherichia coli is transcribed from seven promoters which cover a region of 1.2 kilobase pairs of dna (g. sawers and a. böck, j. bacteriol., 171:2485-2498, 1989). the results demonstrated that all promoters were recognized by the major rna polymerase holoenzyme species e sigma 70 in vitro. further corroboration for multiple functional promoters came from hete ...19892670899
toluene degradation by pseudomonas putida f1. nucleotide sequence of the todc1c2bade genes and their expression in escherichia coli.the nucleotide sequence of the todc1c2bade genes which encode the first three enzymes in the catabolism of toluene by pseudomonas putida f1 was determined. the genes encode the three components of the toluene dioxygenase enzyme system: reductasetol (toda), ferredoxintol (todb), and the two subunits of the terminal dioxygenase (todc1c2); (+)-cis-(1s, 2r)-dihydroxy-3-methylcyclohexa-3,5-diene dehydrogenase (todd); and 3-methylcatechol 2,3-dioxygenase (tode). knowledge of the nucleotide sequence of ...19892670929
bacterial aromatic ring-cleavage enzymes are classified into two different gene families.dioxygenases that catalyze the cleavage of the aromatic ring are classified into two groups according to their mode of ring fission. substrates of ring-cleavage dioxygenases usually contain hydroxyl groups on adjacent aromatic carbons, and intradiol enzymes cleave the ring between these two hydroxyl groups. extradiol enzymes in contrast cleave the ring between one hydroxylated carbon and its adjacent nonhydroxylated carbon. in this study, we determined the complete nucleotide sequence of nahc, t ...19892670937
physically associated enzymes produce and metabolize 2-hydroxy-2,4-dienoate, a chemically unstable intermediate formed in catechol metabolism via meta cleavage in pseudomonas putida.the meta-cleavage pathway of catechol is a major mechanism for degradation of aromatic compounds. in this pathway, the aromatic ring of catechol is cleaved by catechol 2,3-dioxygenase and its product, 2-hydroxymuconic semialdehyde, is further metabolized by either a hydrolytic or dehydrogenative route. in the dehydrogenative route, 2-hydroxymuconic semialdehyde is oxidized to the enol form of 4-oxalocrotonate by a dehydrogenase and then further metabolized to acetaldehyde and pyruvate by the act ...19892681159
isolation and characterization of altered plasmids in mutant strains of pseudomonas putida ncib 9816.the ability of p. putida ncib 9816 to grow with naphthalene (nah+) and salicylate (sal+) is correlated with the presence of an 83 kilobase (kb) conjugative plasmid, pdtg1. derivatives of pdtg1 were obtained from cells after exposure to halogenated analogs of naphthalene or salicylate. the selection of mutants having a nah-sal- or a nah-sal+ phenotype could be enhanced by the addition of triphenyltetrazolium chloride to the indicator medium. structurally modified plasmids were characterized by re ...19892684156
putidaredoxin competitively inhibits cytochrome b5-cytochrome p-450cam association: a proposed molecular model for a cytochrome p-450cam electron-transfer complex.cytochrome b5 has been genetically engineered to afford a fluorescent derivative capable of monitoring its association with cytochrome p-450cam from pseudomonas putida [stayton, p. s., fisher, m. t., & sligar, s. g. (1988) j. biol. chem. 263, 13544-13548]. in the mutant cytochrome b5, threonine is replaced by a cysteine at position 65 (t65c) and has been labeled with the environmentally sensitive fluorophore acrylodan. in this paper, the physiological p-450cam reductant putidaredoxin, an fe2s2.c ...19892690937
trichloroethylene degradation by escherichia coli containing the cloned pseudomonas putida f1 toluene dioxygenase genes.toluene dioxygenase from pseudomonas putida f1 has been implicated as an enzyme capable of degrading trichloroethylene. this has now been confirmed with escherichia coli jm109(pdtg601) that contains the structural genes (todc1c2ba) of toluene dioxygenase under the control of the tac promoter. the extent of trichloroethylene degradation by the recombinant organism depended on the cell concentration and the concentration of trichloroethylene. a linear rate of trichloroethylene degradation was obse ...19892694960
microbial enzymes for creatinine assay: a review.a novel metabolic pathway for the degradation of creatinine with n-methylhydantoin, n-carbamoylsarcosine and sarcosine as successive intermediates was found to operate in pseudomonas putida 77 and many other microorganisms. enzymes involved in this pathway were purified from cells of p. putida 77 and characterized. the first step, deimination of creatinine, is catalyzed by cytosine deaminase/creatinine deiminase. the following two steps, ring-opening of n-methylhydantoin and decarbamoylation of ...19892695273
cloning and sequence analysis of the ntra (rpon) gene of pseudomonas putida.the gene encoding a sigma factor ntra (rpon) was cloned from pseudomonas putida by cross-hybridization with a probe containing a part of the corresponding escherichia coli gene. the cloned gene complemented an ntra mutation of e. coli in activation of xyl genes on the tol plasmid. the predicted amino acid (aa) sequence of p. putida ntra (497 aa; mr 56,215) is highly homologous to ntra proteins from azotobacter vinelandii (81.7%), klebsiella pneumoniae (52.6%), and rhizobium meliloti (36.1%). the ...19892695395
mutations in genes downstream of the rpon gene (encoding sigma 54) of klebsiella pneumoniae affect expression from sigma 54-dependent promoters.two open reading frames (orfs), designated orf95 and orf162, downstream of the klebsiella pneumoniae sigma 54 structural gene (rpon) have been sequenced and shown to encode polypeptides of 12 kd and 16 kd, respectively. orfs homologous to orf95 are present downstream of four out of five rpon genes sequenced to date from a range of gram-negative bacteria, and orf162 is also conserved, at least in pseudomonas putida. chromosomal mutations have been created in each gene using a kan cassette and bot ...19892695747
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