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up-promoter mutations in the trpba operon of pseudomonas aeruginosa.in pseudomonas aeruginosa, the operon encoding tryptophan synthase (trpba) is positively regulated by the trpi protein and an intermediate in tryptophan biosynthesis, indoleglycerol phosphate (ingp). a gene fusion in which the trpba promoter directs expression of the pseudomonas putida xyle gene was constructed. by using a p. putida f1 tode mutant carrying this fusion on a plasmid, three cis-acting mutations that increased xyle expression enough to allow the tode strain to grow on toluene were i ...19911904857
ecologically significant effects of pseudomonas putida ppo301(pro103), genetically engineered to degrade 2,4-dichlorophenoxyacetate, on microbial populations and processes in soil.pseudomonas putida ppo301 (pro103), genetically engineered to degrade 2,4-dichlorophenoxyacetate, affected microbial populations and processes in a nonsterile xeric soil. in soil amended with 2,4-dichlorophenoxyacetate (500 micrograms/g soil) and inoculated with ppo301 (pro103), the rate of evolution of carbon dioxide was retarded for approximately 35 days; there was a transient increase in dehydrogenase activity; and the number of fungal propagules decreased below detection after 18 days. in un ...19911954581
microbial metabolism of quinoline and related compounds. v. degradation of 1h-4-oxoquinoline by pseudomonas putida 33/1.a bacterial strain was isolated with the ability to use 1h-4-oxoquinoline as the sole source of carbon, nitrogen and energy. on the basis of its physiological properties, this isolate was classified as pseudomonas putida. 1h-3-hydroxy-4-oxoquinoline, n-formylanthranilic acid, anthranilic acid and catechol were identified as intermediates in the degradation pathway. the latter was further degraded by ortho-cleavage. the enzymatic conversion of 1h-4-oxoquinoline into 1h-3-hydroxy-4-oxoquinoline re ...19901963786
mechanism of action of urocanase. specific 13c-labelling of the prosthetic nad+ and revision of the structure of its adduct with imidazolylpropionate.1. [4-13c]nicotinate was synthesised and used to support the growth of a nicotinate auxotrophic mutant of pseudomonas putida. 13c-nmr spectroscopy of the isolated urocanase confirmed the efficient incorporation of 13c into c4 of the nicotinamide ring of the tightly bound nad+ cofactor. 2. beta-[( 2'-13c]imidazol-4-yl)propionate was synthesised according to known procedures and used for inhibition of the 13c-labelled urocanase. an increase in the absorbance at 330 nm indicated adduct formation be ...19901976515
intermediate and mechanism of hydroxylation of o-iodophenol by salicylate hydroxylase.salicylate hydroxylase [ec 1.14.13.1] from pseudomonas putida catalyzes the hydroxylation of salicylate, and also o-aminophenol, o-nitrophenol, and o-halogenophenols, to catechol. the reactions with these o-substituted phenols comprise oxygenative deamination, denitration, and dehalogenation, respectively. the reaction stoichiometry, as to nadh oxidized, oxygen consumed, and catechol formed, is 2 : 1 : 1, respectively. the mechanisms for the deiodination and oxygenation of o-iodophenol were inve ...19911917904
nucleotide sequence analysis of the pseudomonas putida ppg7 salicylate hydroxylase gene (nahg) and its 3'-flanking region.gene nahg of naphthalene/salicylate catabolic plasmid nah7 encodes a protein of molecular weight 45,000, salicylate hydroxylase. this enzyme catalyzes the formation of catechol from salicylate, a key intermediate in naphthalene catabolism. dna sequence analysis of the 3.1-kilobase hindiii fragment containing the nahg locus reveals an open reading frame (orf) of 1305 base pairs that corresponds to a protein of 434 amino acid residues. the predicted amino acid sequence of salicylate hydroxylase is ...19911993181
primary structure of xylene monooxygenase: similarities to and differences from the alkane hydroxylation system.xylene monooxygenase, encoded by the tol plasmid of pseudomonas putida, catalyzes the oxidation of toluene and xylenes and consists of two different subunits encoded by xyla and xylm. in this study, the complete nucleotide sequences of these genes were determined and the amino acid sequences of the xyla and xylm products were deduced. the xylm sequence had a 25% homology with alkane hydroxylase, which catalyzes the omega-hydroxylation of fatty acids and the terminal hydroxylation of alkanes. the ...19911999388
construction of tracer plasmids for bacillus sphaericus 1593 utilizing the xyle gene from pseudomonas putida.genetically engineered microorganisms (gems) released into the environment must be traceable in order to accurately assess their impact on the area of release. tracer genes other than those that introduce antibiotic resistance are preferred for use in identifying genetically engineered strains. in this study, we describe the construction of a series of tracer plasmids for use in bacillus sphaericus using the xyle gene from the pseudomonas putida tol plasmid. this gene codes for the enzyme catech ...19912002245
determination of biocatalyst consumption in an aminopeptidase process using automated sample preparation and high-performance liquid chromatography.a rapid and sensitive method has been developed for the determination of the biocatalyst consumption in the chemo-enzymic production of optically pure natural and synthetic alpha-h-amino acids. it is based on automated sample preparation from an enzymic reaction mixture, reversed-phase high-performance liquid chromatographic separation, post-column reaction and fluorimetric detection. the assay procedure has been applied to the enzymic conversion of racemic norvaline amide into l-norvaline, cata ...19911939460
metabolism of poly(3-hydroxyalkanoates) (phas) by pseudomonas oleovorans. identification and sequences of genes and function of the encoded proteins in the synthesis and degradation of pha.pseudomonas oleovorans accumulates poly(3-hydroxyalkanoates) (phas) after growth on medium chain length hydrocarbons. large amounts of this polyester are synthesized when cells are grown under nitrogen-limiting conditions. when nitrogen is resupplied in the medium, the accumulated pha is degraded. in this paper, we describe mutants which are defective in the synthesis or in the degradation of pha. these mutants were used to select dna fragments which encode pha polymerases and a pha depolymerase ...19911989978
sequence of the plasmid-encoded catechol 1,2-dioxygenase-expressing gene, pheb, of phenol-degrading pseudomonas sp. strain est1001.phenol monooxygenase (pmo) and catechol 1,2-dioxygenase (c12o), the two first enzymes of the phenol-degradation pathways, are encoded by a 3.4-kb dna fragment cloned from pseudomonas sp. est1001 plasmid dna. we have previously shown that activation of the cloned genes in pseudomonas putida paw85 is controlled by insertion of the 17-kb transposon, tn4652, from the host chromosome into the plasmid carrying these genes [kivisaar et al. plasmid 24 (1990) 25-36]. transcription of the dna encoding pmo ...19912013408
sequence analysis of the pseudomonas sp. strain p51 tcb gene cluster, which encodes metabolism of chlorinated catechols: evidence for specialization of catechol 1,2-dioxygenases for chlorinated substrates.pseudomonas sp. strain p51 contains two gene clusters located on catabolic plasmid pp51 that encode the degradation of chlorinated benzenes. the nucleotide sequence of a 5,499-bp region containing the chlorocatechol-oxidative gene cluster tcbcdef was determined. the sequence contained five large open reading frames, which were all colinear. the functionality of these open reading frames was studied with various escherichia coli expression systems and by analysis of enzyme activities. the first g ...19912013566
fluorometric determination of phenylacetyl-coa ligase from pseudomonas putida: a very sensitive assay for a newly described enzyme.phenylacetyl-coa ligase (amp-forming) from pseudomonas putida is a newly described enzyme (martinez-blanco, h., reglero, a., rodriguez-aparicio, l.b. and luengo, j.m. (1990) j. biol. chem. 265, 7084-7090) specifically involved in the catabolism of phenylacetic acid. this enzyme catalyzes the formation of phenylacetyl-coa in the presence of atp, coa, mg2+ and phenylacetic acid. a rapid method of assaying this enzyme in partially purified preparations has been developed by coupling this reaction w ...19912009287
microbial degradation of the morphine alkaloids. purification and characterization of morphine dehydrogenase from pseudomonas putida m10.the nadp(+)-dependent morphine dehydrogenase that catalyses the oxidation of morphine to morphinone was detected in glucose-grown cells of pseudomonas putida m10. a rapid and reliable purification procedure involving two consecutive affinity chromatography steps on immobilized dyes was developed for purifying the enzyme 1216-fold to electrophoretic homogeneity from p. putida m10. morphine dehydrogenase was found to be a monomer of mr 32,000 and highly specific with regard to substrates, oxidizin ...19912012614
survival in soils of an herbicide-resistant pseudomonas putida strain bearing a recombinant tol plasmid.pseudomonas putida eez15(pww0-eb62) is a phosphinothricin (ppt)-resistant strain with a recombinant tol plasmid which allows the strain to grow on p-ethylbenzoate. the survival of this strain in sterile agricultural soils depends on the physicochemical properties of the soil. the recombinant pww0-eb62 plasmid and its catabolic functions were stable for periods of up to 1 month in bacteria introduced in unamended soils and only conferred selective advantage to the host bacteria without the plasmi ...19912036014
[phage-stable mutants of pseudomonas putida: new types of stability].more than 170 phage-resistant mutants (prm) of the first order of pseudomonas putida strain ppg1 were obtained using newly isolated and previously described bacteriophages specific for this strain. according to the results of analysis of resistance of the mutants to each of 31 phages of ppg1 strain and 8 phages of the ppn strain, the prm strains were distributed into 20 groups. in most cases, the reason for resistance is loss of absorption capacity of bacteria. however, no direct relation betwee ...19912037253
observation of the o-o stretching raman band for cytochrome p-450cam under catalytic conditions.dioxygen stretching (voo) raman band was observed for the oxy form of pseudomonas putida cytochrome p-450 (p-450cam) generated at room temperature under catalytic conditions, that is, in the presence of d-camphor, beta-nadh, putidaredoxin, and putidaredoxin reductase, by using the mixed flow transient raman apparatus. at the same time the visible absorption spectra were monitored for the transient species. it was found that the voo frequency is little altered by binding of putidaredoxin to p-450 ...19912037577
survival and catabolic activity of natural and genetically engineered bacteria in a laboratory-scale activated-sludge unit.the survival of selected naturally occurring and genetically engineered bacteria in a fully functional laboratory-scale activated-sludge unit (asu) was investigated. the effect of the presence of 3-chlorobenzoate (3cb) on the survival of pseudomonas putida uwc1, with or without a chimeric plasmid, pd10, which encodes 3cb catabolism, was determined. p. putida uwc1(pd10) did not enhance 3cb breakdown in the asu, even following inoculation at a high concentration (3 x 10(8) cfu/ml). the emergence o ...19912014987
characterization of the pseudomonas sp. strain p51 gene tcbr, a lysr-type transcriptional activator of the tcbcdef chlorocatechol oxidative operon, and analysis of the regulatory region.plasmid pp51 of pseudomonas sp. strain p51 contains two gene clusters encoding the degradation of chlorinated benzenes, tcbab and tcbcdef. a regulatory gene, tcbr, was located upstream and divergently transcribed from the chlorocatechol oxidative gene cluster tcbcdef. the tcbr gene was characterized by dna sequencing and expression studies with escherichia coli and pet8c and appeared to encode a 32-kda protein. the activity of the tcbr gene product was analyzed in pseudomonas putida kt2442, in w ...19912050630
structure-function relationships of human aromatase cytochrome p-450 using molecular modeling and site-directed mutagenesis.the conversion of androgens to estrogens is catalyzed by an enzyme complex named aromatase, which consists of a form of cytochrome p-450, aromatase cytochrome p-450 (cytochrome p-450arom), and the flavoprotein, nadph-cytochrome p-450 reductase. as a first step toward investigation of the structure-function relationships of cytochrome p-450arom, we have used computer modeling to align the amino acid sequence of cytochrome p-450arom with that of cytochrome p-450cam from pseudomonas putida and thus ...19912050688
molybdenum-dependent degradation of quinoline by pseudomonas putida chin ik and other aerobic bacteria.eighteen different aerobic bacteria were isolated which utilized quinoline as sole source of carbon, nitrogen, and energy. attempts were unsuccessful at isolating anaerobic quinoline-degrading bacteria. the optimal concentration of quinoline for growth was in the range of 2.5 to 5 mm. some organisms excreted 2-hydroxyquinoline as the first intermediate. hydroxylation of quinoline was catalyzed by a dehydrogenase which was induced in the presence of quinoline or 2-hydroxyquinoline. quinoline dehy ...19912059099
a new family of bacterial regulatory proteins.a new family of bacterial regulatory proteins has been identified by sequence similarity. the family contains the repressor of the bacillus subtilis gluconate operon (gntr), the regulators for histidine utilization in pseudomonas putida (hutcpp) and klebsiella aerogenes (hutcka), the repressor (fadr) of fatty acid degradation in escherichia coli, a regulator involved in the conjugal transfer of the broad host range plasmid pij101 (kora), and three proteins of unidentified function in e. coli (ge ...19912060763
divergent evolution of chloroplast-type ferredoxins.the tol plasmid pww0 of pseudomonas putida encodes a set of enzymes required for the oxidation of toluene to krebs cycle intermediates. the structural genes for these enzymes are encoded in two operons which comprise the xylcmabn and xylxyzltegfjqkih genes, respectively. the function of the xylt gene has not yet been identified. the nucleotide sequence of xylt was determined in this study and putative gene product was shown to contain a sequence characteristic for chloroplast-type ferredoxins. t ...19912065785
[plasmid r68.45 and s-a transduction by the temperate bacteriophage 59 of erwinia carotovora 268].temperature bacteriophage 59 of erwinia carotovera 268 had transduced extrachromosomal dna: plasmids of r68.45 and s-a. before plasmid transduction experiments the suitable donor strains of indicator culture erwinia horticola 450 harbouring r68.45 and s-a were created. the frequency of plasmid r68.45 transfer from pseudomonas putida to e. horticola 450-8 by conjugation was equal to 5 x 10(-8) per a donor cell and in the case of s-a--from e. coli c600 for the same recipient cells--was 2 x 10(-6). ...19912067419
[the effect of amino acids in the asparagine family on the aspartate kinase and homoserine dehydrogenase of ethionine-resistant mutants of pseudomonas putida].the activity of aspartate kinase and homoserin dehydrogenase from ethionine resistant mutants pseudomonas putida 25 and 6 have been studied as affected by amino acids from the family of asparagine. they are characterized by a capacity to the surplus synthesis of methionine. it is shown that mutants have negative regulation of the level of activity of the studied enzymes. it is supposed that the mutations (or mutation) could take place which affected properties of enzymes, which participated dire ...19912067422
the structure of iron superoxide dismutase from pseudomonas ovalis complexed with the inhibitor azide.the 2.9 a resolution structure of iron superoxide dismutase (fesod) (ec 1.15.1.1) from pseudomonas ovalis complexed with the inhibitor azide was solved. comparison of this structure with free enzyme shows that the inhibitor is bound at the open coordination position of the iron, with a bond length of 2.0 a. the metal moves by 0.4 a into the trigonal plane to produce an orthogonal geometry at the iron. binding of the inhibitor also causes a movement of the axial ligand (histidine 26) away from th ...19902075185
purification and characterization of s-alkylcysteine alpha,beta-lyase from pseudomonas putida.s-alkylcysteine alpha,beta-lyase [ec 4.4.1.6] was purified to more than 90% homogeneity from the cell extract of pseudomonas putida icr 3640. the enzyme has a molecular weight of about 195,000, and is composed of six subunits identical in molecular weight (37,000). pyridoxal 5'-phosphate is required as a cofactor. the enzyme catalyzes the alpha,beta-elimination of s-methyl-l-cysteine and its analogs such as s-ethyl-l-cysteine, l-djenkolate, se-methyl-dl-selenocysteine, and o-methyl-l-serine. how ...19902081976
degradation of the metal-cyano complex tetracyanonickelate(ii) by cyanide-utilizing bacterial isolates.ten bacterial isolates capable of growth on tetracyanonickelate(ii) [k2[ni(cn)4]] (tcn) as the sole nitrogen source were isolated from soil, freshwater, and sewage sludge enrichments. seven of the 10 were identified as pseudomonads, while the remaining 3 were classified as klebsiella species. a detailed investigation of one isolate, pseudomonas putida bcn3, revealed a rapid growth rate on tcn (generation time, 2 h), with substrate removal and growth occurring in parallel. in addition to tcn, all ...19902082819
cloning and expression of the plasmid-encoded benzene dioxygenase genes from pseudomonas putida ml2.hybridization using heterologous dioxygenase gene probes indicated the presence of the genes encoding the enzyme benzene dioxygenase on a 112 kb plasmid from pseudomonas putida ml2. they were identified as benzene dioxygenase genes (bed abc1c2) by cloning in escherichia coli and analysis of expression by western blotting using antibodies raised to the four polypeptides of purified benzene dioxygenase.19902083838
phylogenetic comparisons of the branched-chain alpha-ketoacid dehydrogenase complex.1. antibodies against the e1b and e2b components of bovine branched-chain alpha-ketoacid (bcka) dehydrogenase (bckad) complex completely inhibited bcka oxidation in mammalian and avian mitochondria. bcka oxidation by salmonid mitochondria was less affected and the enzyme from pseudomonas putida was unaffected. 2. in rodents, anti-e1b e2b igg inhibited oxidation of all three bcka in a similar dose-dependent manner: oxidation of alpha-ketobutyrate and alpha-keto-y-methiolbutyrate was also partiall ...19902085956
microbial metabolism of quinoline and related compounds. vii. quinoline oxidoreductase from pseudomonas putida: a molybdenum-containing enzyme.the quinoline oxidoreductase from pseudomonas putida was purified 50-fold to homogeneity with 21% recovery, using ammonium sulfate precipitation, hydrophobic interaction-, anion exchange-, and gel chromatography. the mr of the native enzyme was calculated to be 300,000 by gel filtration. sds-polyacrylamide gel electrophoresis of the enzyme revealed three protein bands corresponding to mr 85,000, 30,000 and 20,000. the enzyme contained 8 atoms of iron, 8 atoms of acid-labile sulfide, 2 molecules ...19902090161
an upstream xylr- and ihf-induced nucleoprotein complex regulates the sigma 54-dependent pu promoter of tol plasmid.transcription from promoter pu of the upper catabolic operon of the pseudomonas putida tol plasmid which specifies conversion of toluene/xylenes to benzoate/toluates is activated by the tol-encoded regulator xylr protein in the presence of substrates of the catabolic pathway and in conjunction with the sigma 54(ntra)-containing form of rna polymerase. this regulatory circuit was faithfully reproduced in escherichia coli in single copy gene dosage by integrating the corresponding controlling dete ...19912022186
a predicted three-dimensional structure of human cytochrome p450: implications for substrate specificity.a three-dimensional structure for human cytochrome p450ia1 was predicted based on the crystal coordinates of cytochrome p450cam from pseudomonas putida. as there was only 15% residue identity between the two enzymes, additional information was used to establish an accurate sequence alignment that is a prerequisite for model building. twelve representative eukaryotic sequences were aligned and a net prediction of secondary structure was matched against the known alpha-helices and beta-sheets of p ...19911857713
pseudomonas putida kf715 bphabcd operon encoding biphenyl and polychlorinated biphenyl degradation: cloning, analysis, and expression in soil bacteria.we cloned the entire bphabcd genes encoding degradation of biphenyl and polychlorinated biphenyls to benzoate and chlorobenzoates from the chromosomal dna of pseudomonas putida kf715. the nucleotide sequence revealed two open reading frames corresponding to the bphc gene encoding 2,3-dihydroxybiphenyl dioxygenase and the bphd gene encoding 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (ring-meta-cleavage compound) hydrolase.19902105297
primary and secondary structural patterns in eukaryotic cytochrome p-450 families correspond to structures of the helix-rich domain of pseudomonas putida cytochrome p-450cam. indications for a similar overall topology.an extensive sequence analysis of the eukaryotic cytochrome p-450 (p-450) protein families was conducted with a view to identifying conserved regions that might be related to secondary structural features in the pseudomonas putida camphor hydroxylase (p-450cam). all sequences available on-line were collected, classified and aligned within families. distinctively different sequences were chosen from each of seven eukaryotic families, and an unbiased multi-alignment was constructed. profile patter ...19912040297
pseudomonas chromosomal replication origins: a bacterial class distinct from escherichia coli-type origins.the bacterial origins of dna replication have been isolated from pseudomonas aeruginosa and pseudomonas putida. these origins comprise a second class of bacterial origins distinct from enteric-type origins: both origins function in both pseudomonas species, and neither functions in escherichia coli; enteric origins do not function in either pseudomonad. both cloned sequences hybridize to chromosomal fragments that show properties expected of replication origins. these origin plasmids are highly ...19902106132
chlorinated biphenyl mineralization by individual populations and consortia of freshwater bacteria.comparative studies were performed to investigate the contribution of microbial consortia, individual microbial populations, and specific plasmids to chlorinated biphenyl biodegradation among microbial communities from a polychlorinated biphenyl-contaminated freshwater environment. a bacterial consortium, designated lps10, was shown to mineralize 4-chlorobiphenyl (4cb) and dehalogenate 4,4'-dichlorobiphenyl. the lps10 consortium involved three isolates: pseudomonas testosteroni (lps10a), which m ...19902117875
cloning and expression of rat histidase. homology to two bacterial histidases and four phenylalanine ammonia-lyases.histidase (histidine ammonia-lyase, ec 4.3.1.3) catalyzes the deamination of histidine to urocanic acid. apart from phenylalanine ammonia-lyase, which is not expressed in animals, histidase is the only enzyme known to have a dehydroalanine residue in its active site. the amino site precursor and the mechanism of formation of dehydroalanine are not known. as an initial step to determining the precursor of dehydroalanine in histidase, we have isolated a functional cdna clone for histidase from a r ...19902120224
purification, characterization, and structure of pseudobactin 589 a, a siderophore from a plant growth promoting pseudomonas.under conditions of low-iron stress the plant growth promoting bacterium pseudomonas putida 589 (dsm 50202) produced a yellow-green fluorescent iron-binding peptide siderophore, which was designated pseudobactin 589 a and had an affinity constant toward fe3+ of 10(25) at ph 7. protonated pseudobactin 589 a had the molecular formula c54h78o26n15 and a nominal mass spectral molecular mass of 1353 g/mol. its structure was determined by a combination of nuclear magnetic resonance, fast atom bombardm ...19902145034
enzymatic production of l-tryptophan from dl-serine and indole by a coupled reaction of tryptophan synthase and amino acid racemase.enzymatic production of l-tryptophan from dl-serine and indole by a coupled reaction of tryptophan synthase and amino acid racemase was studied. the tryptophan synthase (ec 4.2.1.20) of escherichia coli catalyzed beta-substitution reaction of l-serine into l-tryptophan and the amino acid racemase (ec 5.1.1.10) of pseudomonas putida catalyzed the racemization of d-serine simultaneously in one reactor. under optimal conditions established for l-tryptophan production, a large-scale production of l- ...19902109982
a new family of rsf1010-derived expression and lac-fusion broad-host-range vectors for gram-negative bacteria.a series of broad-host-range expression and lac fusion vectors, based on rsf1010 derivatives, was constructed. the expression vectors contain various promoters (pnm, plac, ptac and ps1) for expression of foreign genes. the efficiency of the promoters was determined in escherichia coli, rhizobium meliloti, rhizobium leguminosarum and pseudomonas putida by beta-galactosidase activity measurements. of the promoters assayed in e. coli, the most effective is the tac promoter, whereas in soil bacteria ...19902115488
use of a novel cassette to label phenotypically a cryptic plasmid of bacillus subtilis and map loci involved in its stable maintenance.in order to facilitate studies on the maintenance of cryptic plasmids from gram-positive bacteria we have constructed a novel cassette capg1000 (5.0 kb) which carries both a selectable marker (chloramphenicol resistance from staphylococcus aureus plasmid pc194) and a screenable marker (the xyle gene from the tol plasmid of pseudomonas putida expressed from a cloned promoter of bacillus phage spo2) and which is flanked by terminators to prevent transcription from the cassette activating or inhibi ...19902116498
identification and localization of 3-phenylcatechol dioxygenase and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase genes of pseudomonas putida and expression in escherichia coli.the bphc and bphd genes of pseudomonas putida involved in the catabolism of polychlorinated biphenyls or biphenyl were identified, localized, and studied for expression in escherichia coli. this was achieved by cloning a 2.4-kilobase (kb) dna fragment of recombinant cosmid poh101 into hindiii site of puc plasmids downstream of a lacz promoter and measuring the enzyme activities of 3-phenylcatechol dioxygenase (3-pdase; a product of bphc) and the meta-cleavage product 2-hydroxy-6-oxo-6-phenylhexa ...19902160220
complete amino acid sequence of the cytochrome subunit and amino-terminal sequence of the flavin subunit of flavocytochrome c (sulfide dehydrogenase) from chlorobium thiosulfatophilum.the complete amino acid sequence of the 86-residue heme subunit of flavocytochrome c (sulfide dehydrogenase) from the green phototrophic bacterium chlorobium thiosulfatophilum strain tassajara has been determined as follows: apeqsksiprgeilslscagchgtdgksesiiptiygrsaeyiesalldfksga- rpstvmgrhakgysdeeihqiaeyfgslstmnn. the subunit has a single heme-binding site near the n terminus, consisting of a pair of cysteine residues at positions 18 and 21. the out-of-plane ligands are apparently contributed by ...19902161842
physical map of the aromatic amine and m-toluate catabolic plasmid ptdn1 in pseudomonas putida: location of a unique meta-cleavage pathway.a restriction endonuclease map was derived for the aromatic amine and m-toluate catabolic plasmid ptdn1 present in pseudomonas putida ucc22, a derivative of p. putida mt-2. the plasmid is 79 +/- 1 kbp in size and can be divided into a restriction-site-deficient region of 51 +/- 1 kbp and a restriction-site-profuse region of 28 kbp which begins and ends with directly repeating sequences of at least 2 kbp in length. a mutant plasmid isolated after growth of the host on benzoate had lost the restri ...19902168927
loss of tdn catabolic genes by deletion from and curing of plasmid ptdn1 in pseudomonas putida: rate and mode of loss are substrate and ph dependent.the ability to degrade aromatic amines and m-toluate (tdn+ phenotype), encoded by plasmid ptdn1, was lost from pseudomonas putida hosts after subculture in benzoate, succinate, acetate and glucose minimal medium, the fastest rate of loss occurring where benzoate was the substrate. tdn- cells had either lost the entire ptdn1 plasmid or suffered a recombinational deletion of a specific 26 kbp region. proportional increase of tdn- cells resulted from their growth-rate advantage, and additionally, w ...19902168928
l-pipecolic acid metabolism in human liver: l-alpha-aminoadipate delta-semialdehyde oxidoreductase.a soluble enzyme that catalyzes the oxidation of l-alpha-aminoadipate delta-semialdehyde to l-alpha-aminoadipic acid in the presence of nad+ has been isolated and characterized from human liver. this enzyme l-alpha-aminoadipic delta-semialdehyde oxidoreductase has been found to be localized in the cytosol using subcellular fractionation and marker enzyme assays. the reaction product of this enzyme has been identified as l-alpha-aminoadipic acid by use of an amino acid analyzer and thin layer chr ...19902160277
xanthine dehydrogenase and 2-furoyl-coenzyme a dehydrogenase from pseudomonas putida fu1: two molybdenum-containing dehydrogenases of novel structural composition.the constitutive xanthine dehydrogenase and the inducible 2-furoyl-coenzyme a (coa) dehydrogenase could be labeled with [185w]tungstate. this labeling was used as a reporter to purify both labile proteins. the radioactivity cochromatographed predominantly with the residual enzymatic activity of both enzymes during the first purification steps. both radioactive proteins were separated and purified to homogeneity. antibodies raised against the larger protein also exhibited cross-reactivity toward ...19902170335
structure of the dnaa region of micrococcus luteus: conservation and variations among eubacteria.a phylogenetic tree constructed by 5s rrna analysis is composed of three major branches in eubacteria: high g + c gram+, low g + c gram+ and gram- [hori and osawa, mol. biol. evol. 4 (1987) 445-472]. we have shown that the characteristic dnaa region is common among escherichia coli (gram-), pseudomonas putida (gram-), and bacillus subtilis (low g + c gram+). we have now determined the structure of the dnaa region of micrococcus luteus, as a representative of the last branch, high g + c gram+. th ...19902172090
genetic analysis of chromosomal operons involved in degradation of aromatic hydrocarbons in pseudomonas putida tmb.the catabolic pathway for the degradation of aromatic hydrocarbons encoded by pseudomonas putida tmb differs from the tol plasmid-encoded pathway as far as regulation of the upper pathway is concerned. we found, by analyzing tn5-induced mutants and by southern blot hybridization with appropriate probes derived from the tol plasmid pww0, that the catabolic genes of strain tmb were located on the bacterial chromosome and not on the 84-kb plasmid harbored by this strain. the catabolic genes of tmb ...19902172213
transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria.a simple procedure for cloning and stable insertion of foreign genes into the chromosomes of gram-negative eubacteria was developed by combining in two sets of plasmids (i) the transposition features of tn10 and tn5; (ii) the resistances to the herbicide bialaphos, to mercuric salts and organomercurial compounds, and to arsenite, and (iii) the suicide delivery properties of the r6k-based plasmid pgp704. the resulting constructions contained unique noti or sfii sites internal to either the tn10 o ...19902172216
isolation of high frequency of recombination donors from tn5 chromosomal mutants of pseudomonas putida ppn and recalibration of the genetic map.a tn5 loaded derivative of the incp-10 plasmid r91-5 (pmo75) was used as a suicide vector to generate random chromosomal insertion mutations in pseudomonas putida ppn. reintroduction of pmo75 into such mutants resulted in integration of the plasmid at the site of tn5 insertion, giving rise to two classes of high frequency of donors recombination (hfr) donors, transferring chromosome at high frequency (greater than 10(-1) per donor cell) in opposite directions. consequently, tn5 induced auxotroph ...19902174392
upstream regulatory sequence for transcriptional activator xylr in the first operon of xylene metabolism on the tol plasmid.transcription of the first operon coding for m-xylene-degrading enzymes on the tol plasmid of pseudomonas putida is activated by the xylr gene product in the presence of m-xylene. the operon has the consensus sequence of the ntr/nif promoters at -24 and -12 regions, and the transcription is dependent on an rna polymerase containing a sigma factor ntra (rpon or sigma 54). deletion derivatives of the upstream sequence of the operon promoter were made in vitro and connected with the xyle gene on a ...19902174974
ofloxacin versus vancomycin/polymyxin for prevention of infections in granulocytopenic patients.the efficacy and safety of oral ofloxacin were compared with those of vancomycin/polymyxin for prophylaxis of bacterial infections in granulocytopenic patients undergoing chemotherapy for hematologic malignancy.19902153006
putidaredoxin reductase and putidaredoxin. cloning, sequence determination, and heterologous expression of the proteins.the oxidation of camphor by cytochrome p-450cam requires the participation of a flavoprotein, putidaredoxin reductase, and an iron-sulfur protein, putidaredoxin, to mediate the transfer of electrons from nadh to p-450 for oxygen activation. a 2.2-kilobase pair bamhi-stui fragment from whole cell dna of camphor-grown pseudomonas putida has been cloned and sequenced. translation of the sequence revealed two open reading frames that could code for putidaredoxin reductase and putidaredoxin. in the c ...19902180940
the meta cleavage operon of tol degradative plasmid pww0 comprises 13 genes.the meta-cleavage operon of tol plasmid pww0 of pseudomonas putida encodes a set of enzymes which transform benzoate/toluates to krebs cycle intermediates via extradiol (meta-) cleavage of (methyl)catechol. the genetic organization of the operon was characterized by cloning of the meta-cleavage genes into an expression vector and identification of their products in escherichia coli maxicells. this analysis showed that the meta-cleavage operon contains 13 genes whose order and products (in kiloda ...19902183008
nucleotide sequence of the gyrb gene of pseudomonas putida. 19902170947
organization and sequence analysis of the 2,4-dichlorophenol hydroxylase and dichlorocatechol oxidative operons of plasmid pjp4.growth of alcaligenes eutrophus jmp134 on 2,4-dichlorophenoxyacetate requires a 2,4-dichlorphenol hydroxylase encoded by gene tfdb. catabolism of either 2,4-dichlorophenoxyacetate or 3-chlorobenzoate involves enzymes encoded by the chlorocatechol oxidative operon consisting of tfdcdef, which converts 3-chloro- and 3,5-dichlorocatechol to maleylacetate and chloromaleylacetate, respectively. transposon mutagenesis has localized tfdb and tfdcdef to ecori fragment b of plasmid pjp4 (r. h. don, a. j. ...19902185214
a family of positive regulators related to the pseudomonas putida tol plasmid xyls and the escherichia coli arac activators.the xyls family consists of a least 8 different transcriptional regulators. six of these proteins are positive regulators for the catabolism of carbon sources (benzoate and sugars) in escherichia coli, pseudomonas putida and erwinia carotovora, and two of them are involved in pathogenesis in escherichia coli and yersinia enterocolitica. based on protein alignments, the members of this family exhibit a long stretch of homology at the c-terminal end. the regulators involved in the catabolism of ca ...19902186376
identification and characterization of genes for a second anthranilate synthase in pseudomonas aeruginosa: interchangeability of the two anthranilate synthases and evolutionary implications.two anthranilate synthase gene pairs have been identified in pseudomonas aeruginosa. they were cloned, sequenced, inactivated in vitro by insertion of an antibiotic resistance gene, and returned to p. aeruginosa, replacing the wild-type gene. one anthranilate synthase enzyme participates in tryptophan synthesis; its genes are designated trpe and trpg. the other anthranilate synthase enzyme, encoded by phna and phnb, participates in the synthesis of pyocyanin, the characteristic phenazine pigment ...19902153661
microcosm for assessing survival of genetically engineered microorganisms in aquatic environments.laboratory-contained microcosms are important for studying the fate and survival of genetically engineered microorganisms. in this study, we describe a simple aquatic microcosm that utilizes survival chambers in a flowthrough or static renewal system. the model was used to study the survival of genetically engineered and wild-type strains of escherichia coli and pseudomonas putida in the lake water environment. temperature-dependent studies indicated that the genetically engineered microorganism ...19902187407
mutations leading to constitutive expression from the tol plasmid meta-cleavage pathway operon are located at the c-terminal end of the positive regulator protein xyls.the xyls protein is the positive activator of the tol plasmid meta-cleavage pathway operon for the metabolism of alkylbenzoates in pseudomonas putida. the regulator stimulates transcription from the tol meta pathway operon promoter (pm) when activated by benzoate effectors or in the absence of effectors when overproduced. xyls mutant alleles that encode regulators which constitutively mediate expression from pm were isolated and characterized. the mutant proteins all exhibit single amino acid su ...19902193914
design of an enzymatic hybrid system: a useful strategy for the biosynthesis of benzylpenicillin in vitro.a hybrid (prokaryotic-eukaryotic) enzyme system leading to the production of benzylpenicillin has been developed. in vitro synthesis of penicillin g was achieved by incubating 6-aminopenicillanic acid, coa, phenylacetic acid, homogeneously pure phenylacetyl-coa ligase (pa-coa ligase) from pseudomonas putida and acyl-coa:6-apa acyltransferase (at) from penicillium chrysogenum. benzylpenicillin was also obtained when at was coupled with pa-coa ligase and isopenicillin n-synthetase (ipns). this is ...19902178138
evaluation of autoscan-w/a automated microbiology system for the identification of non-glucose-fermenting gram-negative bacilli.we evaluated the ability of the autoscan-w/a (microscan division, baxter healthcare corporation, west sacramento, calif.), in conjunction with the dried colorimetric neg id type 2 panel (dcp) and new rapid fluorometric neg id panel (rfp), to identify non-glucose-fermenting gram-negative bacilli by challenging the system with 310 previously identified reference strains. of these 310 isolates, 286 organisms were in the dcp data base and 269 were in the rfp data base. use of the dcp panels resulted ...19902199522
cloning and sequence analysis of the genes encoding the alpha and beta subunits of the e1 component of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus.a 4175-bp ecori fragment of dna that encodes the alpha and beta chains of the pyruvate dehydrogenase (lipoamide) component (e1) of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus has been cloned in escherichia coli. its nucleotide sequence was determined. open reading frames (pdha, pdhb) corresponding to the e1 alpha subunit (368 amino acids, mr 41,312, without the initiating methionine residue) and e1 beta subunit (324 amino acids, mr 35,306, without the initiating ...19902200674
characterization of the multiple catalytic activities of tartrate dehydrogenase.tartrate dehydrogenase (tdh) has been purified to apparent homogeneity from pseudomonas putida and has been demonstrated to catalyze three different nad(+)-dependent reactions. tdh catalyzes the oxidation of (+)-tartrate to form oxaloglycolate and the oxidative decarboxylation of d-malate to form pyruvate and co2. d-glycerate and co2 are formed from meso-tartrate in a reaction that is formally a decarboxylation with no net oxidation or reduction. the steady-state kinetics of the first two reacti ...19902184888
nucleotide sequence of the gene encoding the repressor for the histidine utilization genes of pseudomonas putida.the hutc gene of pseudomonas putida encodes a repressor which, in combination with the inducer urocanate, regulates expression of the five structural genes necessary for conversion of histidine to glutamate, ammonia, and formate. the nucleotide sequence of the hutc region was determined and found to contain two open reading frames which overlapped by one nucleotide. the first open reading frame (orf1) appeared to encode a 27,648-dalton protein of 248 amino acids whose sequence strongly resembled ...19902203753
nucleotide sequence of the gene encoding the repressor for the histidine utilization genes of klebsiella aerogenes.the hutc gene of klebsiella aerogenes encodes a repressor that regulates expression of the histidine utilization (hut) operons. the dna sequence of a region known to contain hutc was determined and shown to contain two long rightward-reading open reading frames (orfs). one of these orfs was identified as the 3' portion of the hutg gene. the other orf was the hutc gene. the repressor predicted from the hutc sequence contained a helix-turn-helix motif strongly similar to that seen in other dna-bin ...19902203754
sequence and analysis of the rpon sigma factor gene of rhizobium sp. strain ngr234, a primary coregulator of symbiosis.we report the nucleotide sequence of the rpon gene from broad-host-range rhizobium sp. strain ngr234 and analyze the encoded rpon protein, a sigma factor. comparative analysis of the deduced amino acid sequence of rpon from ngr234 with sequences from other gram-negative bacteria identified a perfectly conserved rpon box unique to rpon sigma factors. symbiotic regulatory phenotypes were defined for a site-directed internal deletion within the coding sequence of the rpon gene of rhizobium strain n ...19902211497
transcriptional analysis of the promoter region of the pseudomonas putida branched-chain keto acid dehydrogenase operon.branched-chain keto acid dehydrogenase is a multienzyme complex produced by pseudomonas putida when it is grown in a minimal medium containing branched-chain amino acids. a 1.87-kilobase (kb) dna fragment was cloned and sequenced which contained 0.24 kb of the e1 alpha structural gene and 1.6 kb of upstream dna. there were 854 base pairs (bp) of noncoding dna upstream of bkda1, the first gene of the bkd operon, and 592 bp between the transcriptional and translational starts. the g + c content of ...19902211503
electroporation and expression of plasmid pbr322 in klebsiella aerogenes nctc 418 and plasmid prk2501 in pseudomonas putida cym 318.klebsiella aerogenes nctc 418 and pseudomonas putida cym 318 were transformed via high-voltage electroporation with plasmids pbr322 and prk2501, respectively. the number of transformants obtained was dependent on the applied voltage, capacitance, and cell recovery procedure. for example, 7.87 x 10(4) transformants/micrograms dna were obtained at 2500 v, 25 muf when k. aerogenes cells were electroporated with pbr322 dna. a lower voltage (1500) and capacitance (3 muf) yielded 2.4 x 10(3) transform ...19902187074
purification and characterisation of tol plasmid-encoded benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase of pseudomonas putida.benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase, two enzymes of the xylene degradative pathway encoded by the plasmid tol of a gram-negative bacterium pseudomonas putida, were purified and characterized. benzyl alcohol dehydrogenase catalyses the oxidation of benzyl alcohol to benzaldehyde with the concomitant reduction of nad+; the reaction is reversible. benzaldehyde dehydrogenase catalyses the oxidation of benzaldehyde to benzoic acid with the concomitant reduction of nad+; the re ...19902202600
different types of formaldehyde-oxidizing dehydrogenases in nocardia species 239: purification and characterization of an nad-dependent aldehyde dehydrogenase.three different dehydrogenases able to oxidize formaldehyde were found in the gram-positive methylotroph, nocardia sp. 239: an nad-dependent aldehyde dehydrogenase (na-adh), and nad- and factor-dependent formaldehyde dehydrogenase (fd-fdh), and a dye-linked aldehyde dehydrogenase (dl-adh). the ratio of the activities observed for the two nad-linked enzymes varied with growth conditions: batch-wise grown cells had nearly the same activities for both enzymes; in fed batch-wise grown cells (methano ...19902241149
[studies on respiratory infections in primary care clinic (iii). distribution of bacteria isolated from patients with respiratory infections visiting 21 private clinics in the tohoku district of japan].the bacteriology of the isolates from the throat swab and the sputum respectively of 2,539 patients with respiratory infections visiting 21 private clinics in tohoku district of japan during the period from january to april in 1989 was documented. of the 2,539 patients, 1,694 had an acute upper respiratory infection, 609 had acute bronchitis, 46 had acute pneumonia, 84 had acute exacerbation of chronic respiratory infections and 106 had respiratory infections without diagnosis registered. 1887 ( ...19902243193
nucleotide sequence and expression of the isoamylase gene from an isoamylase-hyperproducing mutant, pseudomonas amyloderamosa jd210.the isoamylase gene (iso) of pseudomonas amyloderamosa jd210, an isoamylase-hyperproducing mutant, was cloned in an isoamylase-deficient and transformable mutant strain k31. by deletion analysis, the iso gene was found to be located within a 3.3 kilobases bamhi fragment. its nucleotide sequence contained an open reading frame of 2328 nucleotides (776 amino acids) encoding a secreted isoamylase precursor. the iso gene fragment was inserted into plasmids pkt230 and pbr 322 in opposite orientations ...19902248978
[the preparation and properties of catechol-1,2-dioxygenase from pseudomonas putida].catechol-1,2-dioxygenase (ec 1.13.11.1) catalyzes the degradation of catechol to cis, cis-muconic acid. the biochemical properties of catechol-1,2-dioxygenase from pseudomonas putida 84103 were investigated. the optimum ph and temperature is 7.5-8.0 and 25-30 degrees c, respectively. cu2+, zn2+ inhibit the enzyme activity. the paper chromatograph and uv absorption spectrum of enzymatic reaction product are accordance with those of the standard muconic acid.19902251833
growth-phase-dependent expression of the pseudomonas putida tol plasmid pww0 catabolic genes.pseudomonas putida tol plasmid pww0 catabolic genes are clustered into two operons. the first, the upper operon, is controlled by the xylr regulatory gene, whereas the second, the meta operon, is controlled by the xyls regulatory gene. the xyls gene itself is subjected to control by xylr. in this study, we show that the tol catabolic operons were poorly induced in cells growing at the early-exponential-growth phase but strongly induced in cells at late-exponential-growth phase. we constructed fu ...19902254244
selection of independent plasmids determining phenol degradation in pseudomonas putida and the cloning and expression of genes encoding phenol monooxygenase and catechol 1,2-dioxygenase.long-term cultivation of the pseudomonas putida multiplasmid strain est1020 on phenol resulted in the formation of individual phe plasmids determining phenol degradation. four types of phe plasmids, pest1024, pest1026, pest1028, and pest1029, are characterized. they all contain a transferrable replicon similar to pwwo-8 with a partly duplicated dna sequence of the 17-kb transposable element of this plasmid and include various amounts of dna that carry genes encoding phenol degradation (phe genes ...19902270227
the 2.1-a resolution structure of iron superoxide dismutase from pseudomonas ovalis.the 2.1-a resolution crystal structure of native uncomplexed iron superoxide dismutase (ec 1.15.1.1) from pseudomonas ovalis was solved and refined to a final r factor of 24%. the dimeric structure contains one catalytic iron center per monomer with an asymmetric trigonal-bipyramidal coordination of protein ligands to the metal. each monomer contains two domains, with the trigonal ligands (histidines 74 and 160; aspartate 156) contributed by the large domain and stabilized by an extended hydroge ...19902271564
mandelate pathway of pseudomonas putida: sequence relationships involving mandelate racemase, (s)-mandelate dehydrogenase, and benzoylformate decarboxylase and expression of benzoylformate decarboxylase in escherichia coli.the genes that encode the five known enzymes of the mandelate pathway of pseudomonas putida (atcc 12633), mandelate racemase (mdla), (s)-mandelate dehydrogenase (mdlb), benzoylformate decarboxylase (mdlc), nad(+)-dependent benzaldehyde dehydrogenase (mdld), and nadp(+)-dependent benzaldehyde dehydrogenase (mdle), have been cloned. the genes for (s)-mandelate dehydrogenase and benzoylformate decarboxylase have been sequenced; these genes and that for mandelate racemase [ransom, s. c., gerlt, j. a ...19902271624
cloning and expression of the ponb gene, encoding penicillin-binding protein 1b of escherichia coli, in heterologous systems.a fragment from the ponb region of the escherichia coli chromosome comprising the promoterless sequence encoding penicillin-binding protein 1b (pbp 1b) has been cloned in a broad-host-range expression vector under the control of the kanamycin resistance gene promoter present in the vector. the hybrid plasmid (pjp3) was used to transform appropriate strains of salmonella typhimurium, pseudomonas putida, and pseudomonas aeruginosa. in all instances, the coding sequence was expressed in the heterol ...19902198260
mandelate racemase and muconate lactonizing enzyme are mechanistically distinct and structurally homologous.mandelate racemase (mr) and muconate lactonizing enzyme (mle) catalyse separate and mechanistically distinct reactions necessary for the catabolism of aromatic acids by pseudomonas putida. the x-ray crystal structure of mr, solved at 2.5 a resolution, reveals that the secondary, tertiary and quaternary structures of mr and mle are remarkably similar; also, mr and mle are about 26% identical in primary structure. however, mr has no detectable mle activity and vice versa. thus, mr and mle constitu ...19902215699
functional modification of an arginine residue on salicylate hydroxylase.salicylate hydroxylase from pseudomonas putida (ec 1.14.13.1, salicylate, nadh:oxygen oxidoreductase) is an fad-containing monooxygenase, which catalyzes decarboxylative hydroxylation of salicylate to produce catechol in the presence of nadh and o2. by chemical treatment of the enzyme with dicarbonyl reagents, such as glyoxal, the original oxygenase activity was converted to the salicylate-dependent nadh-dehydrogenase activity with free fad as electron acceptor. one of twenty arginine residues o ...19902223838
degradation of 3-chlorobiphenyl by in vivo constructed hybrid pseudomonads.3-chlorobiphenyl-degrading bacteria were obtained from the mating between pseudomonas putida strain bn10 and pseudomonas sp. strain b13. strains such as bn210 resulted from the transfer of the genes coding the enzyme sequence for the degradation of chlorocatechols from b13 into bn10, whereas b13 derivatives such as b131 have acquired the biphenyl degradation sequence from bn10. during growth of the hybrid strains on 3-chlorobiphenyl 90% chloride was released. activities of phenylcatechol 2,3-dio ...19902276606
cis-1,2-dihydroxycyclohexa-3,5-diene (nad) oxidoreductase (cis-benzene dihydrodiol dehydrogenase) from pseudomonas putida ncib 12190. 19902280699
toluene dioxygenase from pseudomonas putida f1. 19902280710
benzene dioxygenase from pseudomonas putida, ml2 (ncib 12190). 19902280718
[cloning genes for biosynthesis of pseudomonas putida tryptophan in escherichia coli cells].the trpe, trpc and trpiba genes of pseudomonas putida were cloned by complementation of the corresponding auxotrophic mutations of escherichia coli using pbr322 as a vector. with the exception of trpe, transcription of all genes in new host takes place under control of their own promoters. expression of the trpd gene linked to trpc was not registered in e. coli. repressible trpc enzyme was synthesized constitutively in e. coli. characteristic regulation of p. putida trpba genes via induction by ...19902283047
comparative in vitro activities of newer quinolones against pseudomonas species and xanthomonas maltophilia isolated from patients with cancer.the in vitro susceptibilities of three pseudomonas species (pseudomonas aeruginosa, pseudomonas putida, and pseudomonas fluorescens) and xanthomonas maltophilia to quinolone antimicrobial agents were determined. several newer agents, particularly pd117558, pd117596, pd127391, sparfloxacin (at-4140), a-56620, and temafloxacin, were active against pseudomonas species. x. maltophilia isolates were generally less susceptible than were pseudomonas isolates but were inhibited by some of the newer quin ...19902285297
[rare initiation codons are regulators of expression of the rpoc gene].translation of the rpoc genes in escherichia coli and salmonella typhimurium is known to start from the gug codon. now, using toeprint analysis we have shown uug to be the initiation codon of the pseudomonas putida rpoc gene. if3 does not seem to proofread initiation at the uug codon. the rpoc genes of p. putida, e. coli, and s. typhimurium, which use rare start codons, have strong sd-domains aggagg (p. p.) and gggag (e. c., s. t.), optimal seven-nucleotide spacing between sd and start codons, a ...19902285427
three isozymes of catechol 1,2-dioxygenase (pyrocatechase), alpha alpha, alpha beta, and beta beta, from pseudomonas arvilla c-1.three isozymes of catechol 1,2-dioxygenase (pyrocatechase) from pseudomonas arvilla c-1 were separated using deae-toyopearl chromatography. the specific activities of each isozyme were similar to one another. the molecular weights of isozymes 1, 2, and 3 were estimated to be approximately 67,000, 64,000, and 59,000, respectively, from gel filtration. on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isozymes 1 and 3 gave a single protein band, corresponding to mr = 32,000 and 30,000, ...19902295613
protein components of a cytochrome p-450 linalool 8-methyl hydroxylase.the cytochrome p-450 heme-thiolate monooxygenases that hydroxylate monoterpene hydrocarbon groups are effective models for the cytochrome p-450 family. we have purified and characterized the three proteins from a p-450-dependent linalool 8-methyl hydroxylase in pseudomonas putida (incognita) strain ppg777. the proteins resemble the camphor 5-exohydroxylase components in chemical and physical properties; however, they show neither immunological cross-reactivity nor catalytic activity in heterogen ...19902295633
dna sequences of genes encoding acinetobacter calcoaceticus protocatechuate 3,4-dioxygenase: evidence indicating shuffling of genes and of dna sequences within genes during their evolutionary divergence.the dna sequence of a 2,391-base-pair hindiii restriction fragment of acinetobacter calcoaceticus dna containing the pcachg genes is reported. the dna sequence reveals that a. calcoaceticus pca genes, encoding enzymes required for protocatechuate metabolism, are arranged in a single transcriptional unit, pcaefdbchg, whereas homologous genes are arranged differently in pseudomonas putida. the pcag and pcah genes represent separate reading frames respectively encoding the alpha and beta subunits o ...19902298704
biotransformation of substituted benzoates to the corresponding cis-diols by an engineered strain of pseudomonas oleovorans producing the tol plasmid-specified enzyme toluate-1,2-dioxygenase.the conversion of substituted benzoates into 1,2-cis-dihydroxycyclohexa-3,5-diene carboxylic acids (cis-diols) was effected by using escherichia coli and pseudomonas recombinants carrying the xylxyz genes originating from the pseudomonas putida mt-2 tol plasmid, thus producing toluate-1,2-dioxygenase. pseudomonas oleovorans gpo12 recombinants readily produced meta- and para-substituted cis-diols, but were limited in their oxidation of ortho-substituted substrates.19902306096
cloning and expression of genes involved in 4-chlorobiphenyl transformation by pseudomonas testosteroni: homology to polychlorobiphenyl-degrading genes in other bacteria.the genes of pseudomonas testosteroni strain b-356, specifying the transformation of 4-chlorobiphenyl (4-cb) into 4-chlorobenzoic acid (4-cba) were cloned into pseudomonas putida kt2440 using a broad-host-range cosmid, ppsa842. of 10,000 clones tested, four were able to transform 4-cb. gas chromatographic and mass spectrometric analysis of the catabolic products from two of the 4-cb-transforming clones carrying the hybrid plasmids, pda1 and pda2, demonstrated that pda1 carried a complete set of ...19902311936
streptomyces promoter-probe plasmids that utilise the xyle gene of pseudomonas putida. 19902315034
isolation and partial characterization of an extradiol non-haem iron dioxygenase which preferentially cleaves 3-methylcatechol.a purification procedure has been developed for an extradiol dioxygenase expressed in escherichia coli, which was originally derived from a pseudomonas putida strain able to grow on toluidine. physical and kinetic properties of the enzyme have been investigated. the enzyme has a subunit mr of 33,500 +/- 2000 by sds/polyacrylamide-gel electrophoresis. gel filtration indicates a molecular mass under non-denaturing conditions of 120,000 +/- 20,000. the n-terminal sequence (35 residues) of the enzym ...19902317207
carbon catabolite regulation of phenylacetyl-coa ligase from pseudomonas putida.phenylacetyl-coa ligase (pa-coa ligase) from p. putida u is a newly described enzyme involved in the aerobic catabolism of phenylacetic acid. the enzyme was specifically induced when p. putida was grown in a chemically defined medium containing phenylacetic acid as the sole carbon source. the induction of pa-coa ligase was delayed by adding easily metabolizable carbon sources to the medium; the effect was more drastic in the presence of glucose. glucose did not cause catabolic inactivation but r ...19902322284
purification and biochemical characterization of phenylacetyl-coa ligase from pseudomonas putida. a specific enzyme for the catabolism of phenylacetic acid.a new enzyme, phenylacetyl-coa ligase (amp-forming) (pa-coa ligase, ec 6.2.1-) involved in the catabolism of phenylacetic acid (paa) in pseudomonas putida is described and characterized. pa-coa ligase was specifically induced by paa when p. putida was grown in a chemically defined medium in which phenylacetic acid was the sole carbon source. hydroxyl, methyl-phenylacetyl derivatives, and other paa close structural molecules did not induce the synthesis of this enzyme and neither did acetic, buty ...19902324116
sequence analysis of the huth gene encoding histidine ammonia-lyase in pseudomonas putida.the complete nucleotide sequence of the huth gene, encoding histidine ammonia-lyase (histidase), in pseudomonas putida atcc 12633 has been determined from the appropriate portions of the hut region that had been cloned into escherichia coli. the resulting dna sequence revealed an open reading frame of 1,530 base pairs, corresponding to a protein subunit of approximate molecular weight 53,600, in the location previously identified for the histidase gene by tn1000 mutagenesis. translation began at ...19902332400
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