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molecular cloning, expression and site-directed mutagenesis of glutathione s-transferase from ochrobactrum anthropi.the gene coding for a novel glutathione s-transferase (gst) has been isolated from the bacterium ochrobactrum anthropi. a pcr fragment of 230 bp was obtained using oligonucleotide primers deduced from n-terminal and 'internal' sequences of the purified enzyme. the gene was obtained by screening of a genomic dna partial library from o. anthropi constructed in pbluescript with a pcr fragment probe. the gene encodes a protein (oagst) of 201 amino acids with a calculated molecular mass of 21738 da. ...19989794797
cloning and molecular analysis of the poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) biosynthesis genes in pseudomonas sp. strain 61-3.two types of polyhydroxyalkanoate (pha) biosynthesis gene loci (phb and pha) of pseudomonas sp. strain 61-3, which produces a blend of poly(3-hydroxybutyrate) [p(3hb)] homopolymer and a random copolymer (poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) [p(3hb-co-3ha]) consisting of 3ha units of 4 to 12 carbon atoms, were cloned and analyzed at the molecular level. in the phb locus, three open reading frames encoding polyhydroxybutyrate (phb) synthase (phbcps), beta-ketothiolase (phbaps), and nadph- ...19989851987
initial reductive reactions in aerobic microbial metabolism of 2,4,6-trinitrotoluene.because of its high electron deficiency, initial microbial transformations of 2,4,6-trinitrotoluene (tnt) are characterized by reductive rather than oxidation reactions. the reduction of the nitro groups seems to be the dominating mechanism, whereas hydrogenation of the aromatic ring, as described for picric acid, appears to be of minor importance. thus, two bacterial strains enriched with tnt as a sole source of nitrogen under aerobic conditions, a gram-negative strain called tnt-8 and a gram-p ...199816349484
identification of hydrocarbon-degrading bacteria in soil by reverse sample genome probing.bacteria with limited genomic cross-hybridization were isolated from soil contaminated with c5+, a mixture of hydrocarbons, and identified by partial 16s rrna sequencing. filters containing denatured genomic dnas were used in a reverse sample genome probe (rsgp) procedure for analysis of the effect of an easily degradable compound (toluene) and a highly recalcitrant compound (dicyclopentadiene [dcpd]) on community composition. hybridization with labeled total-community dna isolated from soil exp ...199816349504
light-mediated nitrite accumulation during denitrification by pseudomonas sp. strain jr12.the effect of light on the denitrifying characteristics of a nonphotosynthetic denitrifier, pseudomonas sp. strain jr12, was examined. already at low light intensities, nitrite accumulated as a result of light inhibition of nitrite but not of nitrate reduction rates. exposure of this bacterium to light caused a photooxidation of cytochrome c, an intermediate electron carrier in its respiratory pathway. photoinhibition of nitrite reduction was reversible, as nitrite reduction rates returned to pr ...199816349525
molecular basis of a bacterial consortium: interspecies catabolism of atrazine.pseudomonas sp. strain adp contains the genes, atza, -b, and -c, that encode three enzymes which metabolize atrazine to cyanuric acid. atrazine-catabolizing pure cultures isolated from around the world contain genes homologous to atza, -b, and -c. the present study was conducted to determine whether the same genes are present in an atrazine-catabolizing bacterial consortium and how the genes and metabolism are subdivided among member species. the consortium contained four or more bacterial speci ...199816349478
ntn genes determining the early steps in the divergent catabolism of 4-nitrotoluene and toluene in pseudomonas sp. strain tw3.pseudomonas sp. strain tw3 is able to oxidatively metabolize 4-nitrotoluene and toluene via a route analogous to the upper pathway of the tol plasmids. we report the sequence and organization of five genes, ntnwcmab*, which are very similar to and in the same order as the xyl operon of tol plasmid pww0 and present evidence that they encode enzymes which are expressed during growth on both 4-nitrotoluene and toluene and are responsible for their oxidation to 4-nitrobenzoate and benzoate, respecti ...19989555884
a gene cluster encoding steps in conversion of naphthalene to gentisate in pseudomonas sp. strain u2.pseudomonas sp. strain u2 was isolated from oil-contaminated soil in venezuela by selective enrichment on naphthalene as the sole carbon source. the genes for naphthalene dioxygenase were cloned from the plasmid dna of strain u2 on an 8.3-kb bamhi fragment. the genes for the naphthalene dioxygenase genes nagaa (for ferredoxin reductase), nagab (for ferredoxin), and nagac and nagad (for the large and small subunits of dioxygenase, respectively) were located by southern hybridizations and by nucle ...19989573207
towards a biocatalyst for (s)-styrene oxide production: characterization of the styrene degradation pathway of pseudomonas sp. strain vlb120.in order to design a biocatalyst for the production of optically pure styrene oxide, an important building block in organic synthesis, the metabolic pathway and molecular biology of styrene degradation in pseudomonas sp. strain vlb120 was investigated. a 5.7-kb xhoi fragment, which contained on the same strand of dna six genes involved in styrene degradation, was isolated from a gene library of this organism in escherichia coli by screening for indigo formation. t7 rna polymerase expression expe ...19989603811
low-frequency horizontal transfer of an element containing the chlorocatechol degradation genes from pseudomonas sp. strain b13 to pseudomonas putida f1 and to indigenous bacteria in laboratory-scale activated-sludge microcosms.the possibilities for low-frequency horizontal transfer of the self-transmissible chlorocatechol degradative genes (clc) from pseudomonas sp. strain b13 were investigated in activated-sludge microcosms. when the clc genes were transferred into an appropriate recipient bacterium such as pseudomonas putida f1, a new metabolic pathway for chlorobenzene degradation was formed by complementation which could be selected for by the addition of mono- or 1, 4-dichlorobenzene (cb). under optimized conditi ...19989603824
the atzabc genes encoding atrazine catabolism are located on a self-transmissible plasmid in pseudomonas sp. strain adp.pseudomonas sp. strain adp initiates atrazine catabolism via three enzymatic steps, encoded by atza, -b, and -c, which yield cyanuric acid, a nitrogen source for many bacteria. in-well lysis, southern hybridization, and plasmid transfer studies indicated that the atza, -b, and -c genes are localized on a 96-kb self-transmissible plasmid, padp-1, in pseudomonas sp. strain adp. high-performance liquid chromatography analyses showed that cyanuric acid degradation was not encoded by padp-1. padp-1 w ...19989603862
utility of gram's stain and efficacy of quantitative cultures for posttraumatic pneumonia: a prospective study.this prospective trial examined the efficacy of using bronchoalveolar lavage (bal) for the diagnosis of pneumonia (pn) and the utility of gram's stain (gs) for dictating empiric therapy.19989605666
construction and use of an ipb dna module to generate pseudomonas strains with constitutive trichloroethene and isopropylbenzene oxidation activity.pseudomonas sp. strain jr1 exhibits trichloroethene (tce) oxidation activity with isopropylbenzene (ipb) as the inducer substrate. we previously reported the genes encoding the first three enzymes of the ipb-degradative pathway (ipba1, ipba2, ipba3, ipba4, ipbb, and ipbc) and identified the initial ipb dioxygenase (ipba1 a2a3a4) as responsible for tce cooxidation (u. pflugmacher, b. averhoff, and g. gottschalk, appl. environ. microbiol. 62:3967-3977, 1996). primer extension analyses revealed mul ...19989647815
cloning of a sphingomonas paucimobilis syk-6 gene encoding a novel oxygenase that cleaves lignin-related biphenyl and characterization of the enzyme.sphingomonas paucimobilis syk-6 transforms 2,2'-dihydroxy-3,3'-dimethoxy-5,5'-dicarboxybiphenyl (ddva), a lignin-related biphenyl compound, to 5-carboxyvanillic acid via 2,2',3-trihydroxy-3'-methoxy-5,5'-dicarboxybiphenyl (oh-ddva) as an intermediate (15). the ring fission of oh-ddva is an essential step in the ddva degradative pathway. a 15-kb ecori fragment isolated from the cosmid library complemented the growth deficiency of a mutant on oh-ddva. subcloning and deletion analysis showed that a ...19989647824
chromosomal integration, tandem amplification, and deamplification in pseudomonas putida f1 of a 105-kilobase genetic element containing the chlorocatechol degradative genes from pseudomonas sp. strain b13.analysis of chlorobenzene-degrading transconjugants of pseudomonas putida f1 which had acquired the genes for chlorocatechol degradation (clc) from pseudomonas sp. strain b13 revealed that the clc gene cluster was present on a 105-kb amplifiable genetic element (named the clc element). in one such transconjugant, p. putida rr22, a total of seven or eight chromosomal copies of the entire genetic element were present when the strain was cultivated on chlorobenzene. chromosomal integrations of the ...19989721270
purification and characterization of the coniferyl aldehyde dehydrogenase from pseudomonas sp. strain hr199 and molecular characterization of the gene.the coniferyl aldehyde dehydrogenase (caldh) of pseudomonas sp. strain hr199 (dsm7063), which catalyzes the nad+-dependent oxidation of coniferyl aldehyde to ferulic acid and which is induced during growth with eugenol as the carbon source, was purified and characterized. the native protein exhibited an apparent molecular mass of 86,000 +/- 5,000 da, and the subunit mass was 49.5 +/- 2.5 kda, indicating an alpha2 structure of the native enzyme. the optimal oxidation of coniferyl aldehyde to feru ...19989721273
pseudomonas sp. strain 273, an aerobic alpha, omega-dichloroalkanedegrading bacterium.a gram-negative, aerobic bacterium was isolated from soil; this bacterium grew in 50% (vol/vol) suspensions of 1,10-dichlorodecane (1,10-dcd) as the sole source of carbon and energy. phenotypic and small-subunit ribosomal rna characterizations identified the organism, designated strain 273, as a member of the genus pseudomonas. after induction with 1,10-dcd, pseudomonas sp. strain 273 released stoichiometric amounts of chloride from c5 to c12 alpha, omega-dichloroalkanes in the presence of oxyge ...19989726906
identification and characterization of is1411, a new insertion sequence which causes transcriptional activation of the phenol degradation genes in pseudomonas putida.a new insertion sequence (is element), is1411, was identified downstream of the phenol degradation genes pheba that originated from plasmid dna of pseudomonas sp. strain est1001. according to sequence analysis, is1411 belongs to a new family of is elements that has recently been named the isl3 family (j. mahillon and m. chandler, microbiol. mol. biol. rev. 62:725-774, 1998). is1411 generates 8-bp duplication of the target dna and carries 24-bp inverted repeats (irs), highly homologous to the irs ...19989765560
int-b13, an unusual site-specific recombinase of the bacteriophage p4 integrase family, is responsible for chromosomal insertion of the 105-kilobase clc element of pseudomonas sp. strain b13.pseudomonas sp. strain b13 carries the clcrabde genes encoding chlorocatechol-degradative enzymes on the self-transmissible 105-kb clc element. the element integrates site and orientation specifically into the chromosomes of various bacterial recipients, with a glycine trna structural gene (glyv) as the integration site. we report here the localization and nucleotide sequence of the integrase gene and the activity of the integrase gene product in mediating site-specific integration. the integras ...19989791097
atzc is a new member of the amidohydrolase protein superfamily and is homologous to other atrazine-metabolizing enzymes.pseudomonas sp. strain adp metabolizes atrazine to cyanuric acid via three plasmid-encoded enzymes, atza, atzb, and atzc. the first enzyme, atza, catalyzes the hydrolytic dechlorination of atrazine, yielding hydroxyatrazine. the second enzyme, atzb, catalyzes hydroxyatrazine deamidation, yielding n-isopropylammelide. in this study, the third gene in the atrazine catabolic pathway, atzc, was cloned from a pseudomonas sp. strain adp cosmid library as a 25-kb ecori dna fragment in escherichia coli. ...19989422605
detoxification of protoanemonin by dienelactone hydrolase.protoanemonin is a toxic metabolite which may be formed during the degradation of some chloroaromatic compounds, such as polychlorinated biphenyls, by natural microbial consortia. we show here that protoanemonin can be transformed by dienelactone hydrolase of pseudomonas sp. strain b13 to cis-acetylacrylate. although similar km values were observed for cis-dienelactone and protoanemonin, the turnover rate of protoanemonin was only 1% that of cis-dienelactone. this indicates that at least this pe ...19989440530
a cold-adapted lipase of an alaskan psychrotroph, pseudomonas sp. strain b11-1: gene cloning and enzyme purification and characterization.a psychrotrophic bacterium producing a cold-adapted lipase upon growth at low temperatures was isolated from alaskan soil and identified as a pseudomonas strain. the lipase gene (lipp) was cloned from the strain and sequenced. the amino acid sequence deduced from the nucleotide sequence of the gene (924 bp) corresponded to a protein of 308 amino acid residues with a molecular weight of 33,714. lipp also has consensus motifs conserved in other cold-adapted lipases, i.e., lipase 2 from antarctic m ...19989464382
biochemical and genetic characterization of trans-2'-carboxybenzalpyruvate hydratase-aldolase from a phenanthrene-degrading nocardioides strain.trans-2'-carboxybenzalpyruvate hydratase-aldolase was purified from a phenanthrene-degrading bacterium, nocardioides sp. strain kp7, and characterized. the purified enzyme was found to have molecular masses of 38 kda by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 113 kda by gel filtration chromatography. thus, the homotrimer of the 38-kda subunit constituted an active enzyme. the km and kcat values of this enzyme for trans-2'-carboxybenzalpyruvate were 50 microm and 13 s(-1), r ...19989473051
genetic and functional analysis of the styrene catabolic cluster of pseudomonas sp. strain y2.the chromosomal region of pseudomonas sp. strain y2 involved in the conversion of styrene to phenylacetate (upper catabolic pathway) has been cloned and sequenced. four catabolic genes, styabcd, and two regulatory genes, stysr, were identified. this gene cluster when transferred to escherichia coli w confers to this phenylacetate-degrading host the ability to grow on styrene as the sole carbon and energy source. genes styabcd are homologous to those encoding the styrene upper catabolic pathway i ...19989495743
enzyme specificity of 2-nitrotoluene 2,3-dioxygenase from pseudomonas sp. strain js42 is determined by the c-terminal region of the alpha subunit of the oxygenase component.biotransformations with recombinant escherichia coli expressing the genes encoding 2-nitrotoluene 2,3-dioxygenase (2ntdo) from pseudomonas sp. strain js42 demonstrated that 2ntdo catalyzes the dihydroxylation and/or monohydroxylation of a wide range of aromatic compounds. extremely high nucleotide and deduced amino acid sequence identity exists between the components from 2ntdo and the corresponding components from 2,4-dinitrotoluene dioxygenase (2,4-dntdo) from burkholderia sp. strain dnt (form ...19989495758
involvement of the terminal oxygenase beta subunit in the biphenyl dioxygenase reactivity pattern toward chlorobiphenyls.biphenyl dioxygenase (bph dox) oxidizes biphenyl on adjacent carbons to generate 2,3-dihydro-2,3-dihydroxybiphenyl in comamonas testosteroni b-356 and in pseudomonas sp. strain lb400. the enzyme comprises a two-subunit (alpha and beta) iron sulfur protein (ispbph), a ferredoxin (ferbph), and a ferredoxin reductase (redbph). b-356 bph dox preferentially catalyzes the oxidation of the double-meta-substituted congener 3,3'-dichlorobiphenyl over the double-para-substituted congener 4,4'-dichlorobiph ...19989811638
gene cloning, sequence analysis, and expression of 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase.the gene encoding 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase (mhpco; ec 1.14.12.4) was cloned by using an oligonucleotide probe corresponding to the n terminus of the enzyme to screen a dna library of pseudomonas sp. ma-1. the gene encodes for a protein of 379 amino acid residues corresponding to a molecular mass of 41.7 kda, the same as that previously estimated for mhpco. mhpco was expressed in escherichia coli and found to have the same properties as the native enzyme from pseudom ...19979207074
stereospecific oxidation of (r)- and (s)-1-indanol by naphthalene dioxygenase from pseudomonas sp. strain ncib 9816-4.a recombinant escherichia coli strain which expresses naphthalene dioxygenase (ndo) from pseudomonas sp. strain ncib 9816-4 oxidized (s)-1-indanol to trans-(1s,3s)-indan-1,3-diol (95.5%) and (r)-3-hydroxy-1-indanone (4.5%). the same cells oxidized (r)-1-indanol to cis-1,3-indandiol (71%), (r)-3-hydroxy-1-indanone (18.2%), and cis-1,2,3-indantriol (10.8%). purified ndo oxidized (s)-1-indenol to both syn- and anti-2,3-dihydroxy-1-indanol.19979143136
cross-regulation of toluene monooxygenases by the transcriptional activators tbmr and tbut.the toluene-3-monooxygenase from burkholderia pickettii pko1 and the toluene/benzene-2-monooxygenase from burkholderia (pseudomonas) sp. strain js150 are distinct enzymes which differ not only in catalytic specificity and substrate range but also in the arrangement and sequence of the genes within the operons that encode the enzymes, tbua1ubva2c and tbmabcdef, respectively. in the present study, we examined the transcriptional activation of the ptbua1 and ptbma promoters by their cognate regulat ...19979293027
chemotaxis of pseudomonas spp. to the polyaromatic hydrocarbon naphthalene.two naphthalene-degrading bacteria, pseudomonas putida g7 and pseudomonas sp. strain ncib 9816-4, were chemotactically attracted to naphthalene in drop assays and modified capillary assays. growth on naphthalene or salicylate induced the chemotactic response. p. putida g7 was also chemotactic to biphenyl; other polyaromatic hydrocarbons that were tested did not appear to be chemoattractants for either pseudomonas strain. strains that were cured of the naphthalene degradation plasmid were not att ...19979327579
comparative in vitro activities of trovafloxacin (cp-99,219) against 221 aerobic and 217 anaerobic bacteria isolated from patients with intra-abdominal infections.four hundred thirty-eight bacteria cultured from specimens of patients with serious intra-abdominal infections were tested by agar dilution against trovafloxacin and other quinolones and antimicrobial agents. trovafloxacin inhibited 435 strains (99.3%) at < or =2 microg/ml. all the quinolones had similar activities against enterobacteriaceae and pseudomonas sp., but trovafloxacin showed superior activities against streptococci, enterococci, and anaerobic organisms. because of its excellent in vi ...19979333074
microbiologically proven bacterial infections in aids.we have reviewed the incidence, type and site of microbiologically proven bacterial infection occurring in 52 patients with the acquired immunodeficiency syndrome (aids) who presented to southmead hospital, bristol between 1990 and 1994. a total of 30 (58%) patients had significant bacterial isolates. the majority of infections were community acquired. overall, more infections were caused by gram-negative organisms but gram-positive organisms predominated in bacteraemia. mycobacterium avium intr ...19979373597
cloning, characterization, and sequence analysis of the clce gene encoding the maleylacetate reductase of pseudomonas sp. strain b13.a 3,167-bp psti fragment of genomic dna from pseudomonas sp. strain b13 was cloned and sequenced. the gene clce consists of 1,059 nucleotides encoding a protein of 352 amino acids with a calculated mass of 37,769 da which showed maleylacetate reductase activity. the protein had significant sequence similarities with the polypeptides encoded by tcbf of pp51 (59.4% identical positions), tfdf of pjp4 (55.1%), and tfte of burkholderia cepacia ac1100 (53.1%). the function of tcbf as maleylacetate red ...19979171435
bacterial dl-2-haloacid dehalogenase from pseudomonas sp. strain 113: gene cloning and structural comparison with d- and l-2-haloacid dehalogenases.dl-2-haloacid dehalogenase from pseudomonas sp. strain 113 (dl-dex) catalyzes the hydrolytic dehalogenation of both d- and l-2-haloalkanoic acids to produce the corresponding l- and d-2-hydroxyalkanoic acids, respectively, with inversion of the c2 configuration. dl-dex is a unique enzyme: it acts on the chiral carbon of the substrate and uses both enantiomers as equivalent substrates. we have isolated and sequenced the gene encoding dl-dex. the open reading frame consists of 921 bp corresponding ...19979209038
heterologous expression of biphenyl dioxygenase-encoding genes from a gram-positive broad-spectrum polychlorinated biphenyl degrader and characterization of chlorobiphenyl oxidation by the gene products.the bpha1a2a3a4 gene cluster, encoding a biphenyl dioxygenase from rhodococcus globerulus p6, a gram-positive microorganism able to degrade a wide spectrum of polychlorobiphenyls (pcbs), was expressed in pseudomonas putida, thereby allowing characterization of chlorobiphenyl oxidation by this enzyme. while p6 biphenyl dioxygenase activity was observed in p. putida containing bpha1a2a3a4, no activity was detected in escherichia coli cells containing the same gene cluster. in e. coli, transcriptio ...19979068637
isolation and characterization of a chlorinated-pyridinol-degrading bacterium.the isolation of a pure culture of bacteria able to use 3,5,6-trichloro-2-pyridinol (tcp) as a sole source of carbon and energy under aerobic conditions was achieved for the first time. the bacterium was identified as a pseudomonas sp. and designated atcc 700113. [2,6-(sup14)c]tcp degradation yielded (sup14)co(inf2), chloride, and unidentified polar metabolites.199716535719
effect of selected polysaccharide-producing soil bacteria on hyperhydricity control in oregano tissue cultures.hyperhydricity, or vitrification, is a physiological malformation affecting tissue culture-generated plants. this malformation is associated with excessive hydration and poor lignification and results in poor regeneration of plants. we have tested hyperhydricity prevention in oregano by several nonspecific polysaccharide-producing rhizosphere bacteria. among these bacteria, pseudomonas mucidolens and another pseudomonas sp. prevented hyperhydricity and improved acclimation of oregano clones. the ...199716535525
dechlorination of atrazine by a rhizobium sp. isolate.a rhizobium sp. strain, named patr, was isolated from an agricultural soil and found to actively degrade the herbicide atrazine. incubation of patr in a basal liquid medium containing 30 mg of atrazine liter(sup-1) resulted in the rapid consumption of the herbicide and the accumulation of hydroxyatrazine as the only metabolite detected after 8 days of culture. experiments performed with ring-labeled [(sup14)c]atrazine indicated no mineralization. the enzyme responsible for the hydroxylation of a ...199716535552
identification and characterization of genes encoding carbazole 1,9a-dioxygenase in pseudomonas sp. strain ca10.nucleotide sequence analysis of the flanking regions of the carbc genes of pseudomonas sp. strain ca10 revealed that there were two open reading frames (orfs) orf4 and orf5, in the upstream region of carbc. similarly, three orfs, orf6 to orf8, were found in the downstream region of carbc. the deduced amino acid sequences of orf6 and orf8 showed homologies with ferredoxin and ferredoxin reductase components of bacterial multicomponent dioxygenase systems, respectively. orf4 and orf5 had the same ...19979244274
purification of an extracellular d-(-)-3-hydroxybutyrate oligomer hydrolase from pseudomonas sp. strain a1 and cloning and sequencing of its gene.an extracellular d-(-)-3-hydroxybutyrate oligomer hydrolase was purified from a poly(3-hydroxybutyrate)-degrading bacterium, pseudomonas sp. strain a1. the purified enzyme hydrolyzed the d-(-)-3-hydroxybutyrate dimer and trimer at similar rates. the enzyme activity was inhibited by a low concentration of diisopropylfluorophosphate. the molecular weight of the hydrolase was estimated to be about 70,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. a 10-kbp dna fragment of a1 was d ...19978981982
the atzb gene of pseudomonas sp. strain adp encodes the second enzyme of a novel atrazine degradation pathway.we previously reported the isolation of a 21.5-kb genomic dna fragment from pseudomonas sp. strain adp, which contains the atza gene, encoding the first metabolic step for the degradation of the herbicide atrazine (m. de souza, l. p. wackett, k. l. boundy-mills, r. t. mandelbaum, and m. j. sadowsky, appl. environ. microbiol. 61:3373-3378, 1995). in this study, we show that this fragment also contained the second gene of the atrazine metabolic pathway, atzb. atzb catalyzed the transformation of h ...19979055410
molecular characterization of genes of pseudomonas sp. strain hr199 involved in bioconversion of vanillin to protocatechuate.the gene loci vdh, vana, and vanb, which are involved in the bioconversion of vanillin to protocatechuate by pseudomonas sp. strain hr199 (dsm 7063), were identified as the structural genes of a novel vanillin dehydrogenase (vdh) and the two subunits of a vanillate demethylase (vana and vanb), respectively. these genes were localized on an ecori fragment (e230), which was cloned from a pseudomonas sp. strain hr199 genomic library in the cosmid pvk100. the vdh gene was identified on a subfragment ...19979098058
enzymology of oxidation of tropic acid to phenylacetic acid in metabolism of atropine by pseudomonas sp. strain at3.pseudomonas sp. strain at3 grew with dl-tropic acid, the aromatic component of the alkaloid atropine, as the sole source of carbon and energy. tropic acid-grown cells rapidly oxidized the growth substrate, phenylacetaldehyde, and phenylacetic acid. crude cell extracts, prepared from dl-tropic acid-grown cells, contained two nad+-linked dehydrogenases which were separated by ion-exchange chromatography and shown to be specific for their respective substrates, dl-tropic acid and phenylacetaldehyde ...19979023182
preparation of gm1 ganglioside with sialidase-producing marine bacteria as a microbial biocatalyst.this paper describes the preparation of monosialoganglioside gm1 with sialidase-producing marine bacteria as a microbial biocatalyst. a new sialidase-producing bacterium, identified tentatively as pseudomonas sp. strain yf-2, was isolated from seawater by enrichment culture with ganglioside as the sole source of carbon. when yf-2 was cultured in a synthetic medium containing crude bovine brain gangliosides at 25 degrees c for 3 days, 80 to 90% of the gangliosides were converted to gm1. gm1 was t ...19979143118
structure and expression of a pyrimidine gene cluster from the extreme thermophile thermus strain zo5.on a 4.7-kbp hindiii clone of thermus strain zo5 dna, complementing an aspartate carbamoyltransferase mutation in escherichia coli, we identified a cluster of four potential open reading frames corresponding to genes pyrr, and pyrb, an unidentified open reading frame named bbc, and gene pyrc. the transcription initiation site was mapped at about 115 nucleotides upstream of the pyrr translation start codon. the cognate thermus pyr promoter also functions in heterologous expression of thermus pyr ...19979171389
multiple pathways for toluene degradation in burkholderia sp. strain js150.burkholderia (pseudomonas) sp. strain js150 uses multiple pathways for the metabolism of catechols that result from degradation of aromatic compounds. this suggests that the strain also uses multiple upstream pathways for the initial hydroxylation of aromatic substrates. two distinct dna fragments that allowed pseudomonas aeruginosa pao1c to grow with benzene as a sole carbon source were cloned from strain js150. one of the recombinant plasmids containing the initial steps for the degradative pa ...19979327568
use of a promoterless lacz gene insertion to investigate chitinase gene expression in the marine bacterium pseudoalteromonas sp. strain s9.sequence data for genes encoding 16s rrna indicated that the marine strain previously named pseudomonas sp. strain s9 would be better identified as a pseudoalteromonas sp. by use of transposon mutagenesis, a chitinase-negative mutant of s9 with a lacz reporter gene insertion was isolated. part of the interrupted gene was cloned and sequenced. the deduced amino acid sequence had homology to sequences of bacterial chitinases. expression of the chitinase gene promoter was quantified by measuring th ...19979251187
cloning of genes involved in carbazole degradation of pseudomonas sp. strain ca10: nucleotide sequences of genes and characterization of meta-cleavage enzymes and hydrolase.the dna fragment encoding meta-cleavage enzymes and the meta-cleavage compound hydrolase, involved in carbazole degradation, was cloned from the carbazole-utilizing bacterium pseudomonas sp. strain ca10. dna sequence analysis of this 2.6-kb smai-sphi fragment revealed that there were three open reading frames (orf1, orf2, and orf3, in this gene order). orf1 and orf2 were indispensable for meta-cleavage activity for 2'-aminobiphenyl-2,3-diol and its easily available analog, 2,3-dihydroxybiphenyl, ...19979244273
cloning, sequencing, and expression of isopropylbenzene degradation genes from pseudomonas sp. strain jr1: identification of isopropylbenzene dioxygenase that mediates trichloroethene oxidation.pseudomonas sp. strain jr1, recently isolated with isopropylbenzene (ipb) as the inducer substrate for trichloroethene (tce) oxidation (b. dabrock, j. riedel, j. bertram, and g. gottschalk, arch. microbiol 158:9-13, 1992), is able to degrade ipb via the meta-cleavage pathway. the genes encoding the first three enzymes in the catabolism of isopropylbenzene were isolated from a genomic library with the broad-host-range cosmid vector pwe15. a 7.6-kb fragment from a 37.7-kb primary cosmid clone was ...19968899984
regio- and stereospecific oxidation of fluorene, dibenzofuran, and dibenzothiophene by naphthalene dioxygenase from pseudomonas sp. strain ncib 9816-4.the regio- and stereospecific oxidation of fluorene, dibenzofuran, and dibenzothiophene was examined with mutant and recombinant strains expressing naphthalene dioxygenase from pseudomonas sp. strain ncib 9816-4. the initial oxidation products were isolated and identified by gas chromatography-mass spectrometry and nuclear magnetic resonance spectrometry. salicylate-induced cells of pseudomonas sp. strain 9816/11 and isopropyl-beta-d-thiogalactopyranoside-induced cells of escherichia coli jm109( ...19968899998
atropine metabolism by pseudomonas sp. strain at3: evidence for nortropine as an intermediate in tropine breakdown and reactions leading to succinate.pseudomonas strain at3, isolated by elective culture with atropine, hydrolyzed atropine and grew diauxically, first on the tropic acid and then on the tropine. tropine was also used as a sole carbon and energy source. the methyl group of tropine was eliminated as formaldehyde, and the nortropine thus formed was a precursor of 6-hydroxycyclohepta-1,4-dione. ammonia was detected as a product of nitrogen elimination. 6-hydroxycyclohepta-1,4-dione was oxidized to cyclohepta-1,3,5-trione by an induce ...199616535398
transformation of low concentrations of 3-chlorobenzoate by pseudomonas sp. strain b13: kinetics and residual concentrations.the transformation of 3-chlorobenzoate (3cb) and acetate at initial concentrations in the wide range of 10 nm to 16 mm was studied in batch experiments with pseudomonas sp. strain b13. transformation rates of 3cb at millimolar concentrations could be described by michaelis-menten kinetics (k(infm), 0.13 mm; v(infmax), 24 nmol (middot) mg of protein(sup-1) (middot) min(sup-1)). experiments with nanomolar and low micromolar concentrations of 3cb indicated the possible existence of two different tr ...199616535232
oxidation of 6,7-dihydro-5h-benzocycloheptene by bacterial strains expressing naphthalene dioxygenase, biphenyl dioxygenase, and toluene dioxygenase yields homochiral monol or cis-diol enantiomers as major products.bacterial strains expressing naphthalene, biphenyl, and toluene dioxygenase were examined for their abilities to oxidize 6,7-dihydro-5h-benzocycloheptene (benzocyclohept-1-ene). the major oxidation products were isolated, and their absolute configurations were determined by chiral 1h nuclear magnetic resonance analysis of diastereomeric boronate esters, chiral stationary-phase high-pressure liquid chromatography, and stereo-chemical correlation. pseudomonas sp. strain 9816/11 and sphingomonas ya ...19968919798
measurement of minimum substrate concentration (smin) in a recycling fermentor and its prediction from the kinetic parameters of pseudomonas strain b13 from batch and chemostat cultures.the minimum substrate concentration required for growth, smin, was measured for pseudomonas sp. strain b13 with 3-chlorobenzoate (3cb) and acetate in a recycling fermentor. the substrates were provided alone or in a mixture. smin values predicted with kinetic parameters from resting-cell batches and chemostat cultures differed clearly from the values measured in the recycling fermentor. when 3cb and acetate were fed as single substrates, the measured smin values were higher than the individual s ...19968967775
comparison of factors influencing trichloroethylene degradation by toluene-oxidizing bacteria.the degradation of trichloroethylene (tce) by toluene-oxidizing bacteria has been extensively studied, and yet the influence of environmental conditions and physiological characteristics of individual strains has received little attention. to consider these effects, the levels of tce degradation by strains distinguishable on the basis of toluene and nitrate metabolism were compared under aerobic or hypoxic conditions in the presence and absence of nitrate and an exogenous electron donor, lactate ...19968975612
catechol 2,3-dioxygenases functional in oxygen-limited (hypoxic) environments.we studied the degradation of toluene for bacteria isolated from hypoxic (i.e., oxygen-limited) petroleum-contaminated aquifers and compared such strains with other toluene degraders. three pseudomonas isolates, p. pickettii pko1, pseudomonas sp. strain w31, and p. fluorescens cfs215, grew on toluene when nitrate was present as an alternate electron acceptor in hypoxic environments. we examined kinetic parameters (k(m) and vmax) for catechol 2,3-dioxygenase (c230), a key shared enzyme of the tol ...19968633871
stereospecific dihydroxylation of the styrene vinyl group by purified naphthalene dioxygenase from pseudomonas sp. strain ncib 9816-4.naphthalene dioxygenase (ndo) from pseudomonas sp. strain ncib 9816-4 adds both atoms of the dioxygen molecule to styrene to form (r)-l-phenyl-1,2-ethanediol. product formation is tightly coupled to dioxygen consumption and nadh oxidation. ndo oxidizes styrene-d8 at almost the same initial rate as styrene. the results indicate that dioxygen activation by ndo is different from that by cytochrome p-450 and other monooxygenases, which oxidize styrene to styrene 1,2-oxide.19968655521
chloroform mineralization by toluene-oxidizing bacteria.seven toluene-oxidizing bacterial strains (pseudomonas mendocina kr1, burkholderia cepacia g4, pseudomonas putida f1, pseudomonas pickettii pko1, and pseudomonas sp. strains envpc5, envbf1, and env113) were tested for their ability to degrade chloroform (cf). the greatest rate of cf oxidation was achieved with strain envbf1 (1.9 nmol/min/mg of cell protein). cf also was oxidized by p. mendocina kr1 (0.48 nmol/min/mg of cell protein), strain envpc5 (0.49 nmol/min/mg of cell protein), and escheric ...19968702263
genetic structures of the genes encoding 2,3-dihydroxybiphenyl 1,2-dioxygenase and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid hydrolase from biphenyl- and 4-chlorobiphenyl-degrading pseudomonas sp. strain dj-12.the pcbc and pcbd genes of pseudomonas sp. strain dj-12, a natural isolate degrading biphenyl and 4-chlorobiphenyl, encode the 2,3-dihydroxybiphenyl 1,2-dioxygenase and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid hydrolase, respectively. the two genes were sequenced and appear to be present in the order pcbd-pcbc as an operon.19968572703
atrazine chlorohydrolase from pseudomonas sp. strain adp: gene sequence, enzyme purification, and protein characterization.pseudomonas sp. strain adp metabolizes atrazine to carbon dioxide and ammonia via the intermediate hydroxyatrazine. the genetic potential to produce hydroxyatrazine was previously attributed to a 1.9-kb avai dna fragment from strain adp (m. l. de souza, l. p. wackett, k. l. boundy-mills, r. t. mandelbaum, and m. j. sadowsky, appl. environ. microbiol. 61:3373-3378, 1995). in this study, sequence analysis of the 1.9-kb avai fragment indicated that a single open reading frame, atza, encoded an acti ...19968759853
toluene and ethylbenzene oxidation by purified naphthalene dioxygenase from pseudomonas sp. strain ncib 9816-4.purified naphthalene dioxygenase (ndo) from pseudomonas sp. strain ncib 9816-4 oxidized toluene to benzyl alcohol and benzaldehyde by reactions involving benzylic monooxygenation and dioxygen-dependent alcohol oxidation, respectively. xylene and nitrotoluene isomers were also oxidized to substituted benzyl alcohol and benzaldehyde derivatives. ndo oxidized ethylbenzene sequentially through (s)-1-phenethyl alcohol (77% enantiomeric excess) and acetophenone to 2-hydroxyacetophenone. in addition, n ...19968795196
two types of novel dipeptidyl aminopeptidases from pseudomonas sp. strain wo24.two kinds of dipeptidyl aminopeptidase i (dap i [cathepsin c])-like activities which hydrolyze gly-phe-p-nitroanilide (gly-phe-pna) were detected in pseudomonas sp. strain wo24. they were purified and characterized. the isolated enzymes, named dap bii and dap biii, were revealed to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page) and isoelectric focusing. dap bii was estimated to have a molecular mass of 150,000 da by gel filtration and a subunit size of 73, ...19968892831
a novel dipeptidyl aminopeptidase from pseudomonas sp. strain wo24.an activity similar to that of dipeptidyl aminopeptidase i (dap i) which releases dipeptide from gly-arg-p-nitroanilide (gly-arg-pna) was detected in a pseudomonas sp. an enzyme was isolated and purified about 400-fold by a series of column chromatographies. the enzyme, named dap bi (dap from bacteria, type i), was revealed to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page) and isoelectric focusing. the molecular mass was estimated to be 82 kda by sds-page ...19968631703
maleylacetate reductases in chloroaromatic-degrading bacteria using the modified ortho pathway: comparison of catalytic properties.the maleylacetate reductases from pseudomonas aeruginosa rho1 and alcaligenes eutrophus jmp134 were tested for activity and affinity to various maleylacetates as well as dechlorinating properties. the dechlorinating activity and the kcat/km values revealed high-level similarity of these reductases to that of pseudomonas sp. strain b13.19968550433
phenotypic and genotypic characterization of phenanthrene-degrading fluorescent pseudomonas biovars.a total of 41 phenanthrene degraders were isolated from a former coal gasification site by using pseudomonas-selective gould's s1 medium. all isolates were found to belong to the fluorescent pseudomonas group and were subjected to characterization by phenotypic methods, including classical taxonomic tests, api 20ne, and biolog gn, and the strains were further characterized by the genotypic method repetitive extragenic palindromic pcr (rep-pcr). by using classical tests, the population was found ...19968837438
genetic exchange in soil between introduced chlorobenzoate degraders and indigenous biphenyl degraders.pseudomonas aeruginosa jb2, a chlorobenzoate degrader, was inoculated into soil having indigenous biphenyl degraders but no identifiable 2-chlorobenzoate (2cba) or 2,5-dichlorobenzoate (2,5dcba) degraders. the absence of any indigenous chlorobenzoate degraders was noted by the failure to obtain enrichment cultures with the addition of 2cba, 3cba, or 2,5dcba and by the failure of soil dna to hybridize to the tfdc gene, which encodes ortho fission of chlorocatechols. in contrast, dna extracted fro ...19968837452
dihydroxylation and dechlorination of chlorinated biphenyls by purified biphenyl 2,3-dioxygenase from pseudomonas sp. strain lb400.oxidation of biphenyl and nine chlorinated biphenyls (cbs) by the biphenyl 2,3-dioxygenase from pseudomonas sp. strain lb400 was examined. the purified terminal oxygenase required the addition of partially purified electron transport components, nad(p)h, and ferrous iron to oxidize biphenyl and cbs. cis-biphenyl 2,3-dihydrodiol was produced with biphenyl as the substrate. dihydrodiols were produced from all cbs, and more than one compound was produced with most substrates. catechols were produce ...19958002618
bacterial degradation of m-nitrobenzoic acid.pseudomonas sp. strain js51 grows on m-nitrobenzoate (m-nba) with stoichiometric release of nitrite. m-nba-grown cells oxidized m-nba and protocatechuate but not 3-hydroxybenzoate, 4-hydroxy-3-nitrobenzoate, 4-nitrocatechol, and 1,2,4-benzenetriol. protocatechuate accumulated transiently when succinate-grown cells were transferred to media containing m-nba. respirometric experiments indicated that the conversion of m-nba to protocatechuate required 1 mol of oxygen per mol of substrate. conversio ...19957574625
maleylacetate reductase of pseudomonas sp. strain b13: specificity of substrate conversion and halide elimination.maleylacetate reductase (ec 1.3.1.32) plays a major role in the degradation of chloroaromatic compounds by channelling maleylacetate and some chlorinated derivatives into the 3-oxoadipate pathway. several substituted maleylacetates were prepared in situ by alkaline or enzymatic hydrolysis of dienelactones as the precursor. the conversion of these methyl-, chloro-, fluoro-, and bromo-substituted maleylacetates by malelacetate reductase from 3-chlorobenzoate-grown cells of pseudomonas sp. strain b ...19957814320
characterization of biphenyl catabolic genes of gram-positive polychlorinated biphenyl degrader rhodococcus sp. strain rha1.rhodococcus sp. strain rha1 is a gram-positive polychlorinated biphenyl (pcb) degrader which can degrade 10 ppm of pcb48 (equivalent to aroclor1248), including tri-, tetra-, and pentachlorobiphenyls, in a few days. we isolated the 7.6-kb ecori-bamhi fragment carrying the biphenyl catabolic genes of rha1 and determined their nucleotide sequence. on the basis of deduced amino acid sequence homology, we identified six bph genes, bpha1a2a3a4, bphb, and bphc, that are responsible for the initial thre ...19957793929
lipase modulator protein (liml) of pseudomonas sp. strain 109.plasmids containing a pseudomonas sp. strain 109 extracellular lipase gene (lipl) lacking nh2-terminal sequence and a lipase modulator gene (liml) lacking the nh2-terminal hydrophobic region were constructed and expressed independently in escherichia coli by using the t7 promoter expression vector system. recombinant lipl (rlipl) was produced as inclusion bodies, whereas recombinant liml (rliml) was present as a soluble protein. during in vitro renaturation of the purified rlipl inclusion bodies ...19957868599
aromatic effector activation of the ntrc-like transcriptional regulator phhr limits the catabolic potential of the (methyl)phenol degradative pathway it controls.pseudomonas putida p35x (ncib 9869) metabolizes phenol and monomethylphenols via a chromosomally encoded meta-cleavage pathway. we have recently described a 13.4-kb fragment of the chromosome that codes for the first eight genes of the catabolic pathway and a divergently transcribed positive regulator, phhr. the eight structural genes lie in an operon, the phh operon, downstream of a -24 tggc, -12 ttgc promoter sequence. promoters of this class are recognized by rna polymerase that utilizes the ...19957883704
survival and activity of pseudomonas sp. strain b13(fr1) in a marine microcosm determined by quantitative pcr and an rrna-targeting probe and its effect on the indigenous bacterioplankton.genetically engineered pseudomonas sp. strain b13(fr1) was released into laboratory-scale marine ecosystem models (microcosms). survival of the introduced population in the water column and the sediment was determined by plating on a selective medium and by quantitative competitive pcr. the activity of the released bacteria was determined by in situ hybridization of single cells with a specific rrna-targeting oligonucleotide probe. two microcosms were inoculated with 10(6) cells ml-1, while an u ...19957538271
desaturation, dioxygenation, and monooxygenation reactions catalyzed by naphthalene dioxygenase from pseudomonas sp. strain 9816-4.the stereospecific oxidation of indan and indene was examined with mutant and recombinant strains expressing naphthalene dioxygenase of pseudomonas sp. strain 9816-4. pseudomonas sp. strain 9816/11 and escherichia coli jm109(de3)[pdtg141] oxidized indan to (+)-(1s)-indanol, (+)-cis-(1r,2s)-indandiol, (+)-(1s)-indenol, and 1-indanone. the same strains oxidized indene to (+)-cis-(1r,2s)-indandiol and (+)-(1s)-indenol. purified naphthalene dioxygenase oxidized indan to the same four products formed ...19957751268
cloning, characterization, and expression of a gene region from pseudomonas sp. strain adp involved in the dechlorination of atrazine.we previously identified a pseudomonas sp. strain, adp, which rapidly metabolized atrazine in liquid culture, agar plates, and soils (r. t. mandelbaum, d. l. allan, l. p. wackett, appl. environ. microbiol. 61:1451-1457, 1995). in this study, we report the cloning and partial characterization of a gene region from pseudomonas sp. strain adp that encodes atrazine degradation activity. a 22-kb ecori genomic dna fragment, designated pmd1, was shown to encode atrazine dechlorination activity in esche ...19957574646
transformation of carbon tetrachloride via sulfur and oxygen substitution by pseudomonas sp. strain kc.pseudomonas sp. strain kc transforms carbon tetrachloride into carbon dioxide and nonvolatile products, without chloroform as an intermediate. to define the pathway for hydrolysis, nonvolatile products were analyzed. condensation products containing the carbon atom of carbon tetrachloride as carbonyl and thioxo moieties were identified, indicating the intermediacy of phosgene and thiophosgene in the pathway.19957721711
purification and characterization of the oxygenase component of biphenyl 2,3-dioxygenase from pseudomonas sp. strain lb400.the iron-sulfur protein of biphenyl 2,3-dioxygenase (ispbph) was purified from pseudomonas sp. strain lb400. the protein is composed of a 1:1 ratio of a large (alpha) subunit with an estimated molecular weight of 53,300 and a small (beta) subunit with an estimated molecular weight of 27,300. the native molecular weight was 209,000, indicating that the protein adopts an alpha 3 beta 3 native conformation. measurements of iron and acid-labile sulfide gave 2 mol of each per mol of alpha beta hetero ...19957592331
nucleotide sequence analysis of genes encoding a toluene/benzene-2-monooxygenase from pseudomonas sp. strain js150.it was previously shown by others that pseudomonas sp. strain js150 metabolizes benzene and alkyl- and chloro-substituted benzenes by using dioxygenase-initiated pathways coupled with multiple downstream metabolic pathways to accommodate catechol metabolism. by cloning genes encoding benzene-degradative enzymes, we found that strain js150 also carries genes for a toluene/benzene-2-monooxygenase. the gene cluster encoding a 2-monooxygenase and its cognate regulator was cloned from a plasmid carri ...19957574644
induction of toluene oxidation activity in pseudomonas mendocina kr1 and pseudomonas sp. strain envpc5 by chlorinated solvents and alkanes.toluene oxidation activity in pseudomonas mendocina kr1 and pseudomonas sp. strain envpc5 was induced by trichloroethylene (tce), and induction was followed by the degradation of tce. higher levels of toluene oxidation activity were achieved in the presence of a supplemental growth substrate such as glutamate, with levels of activity of up to 86% of that observed with toluene-induced cells. activity in p. mendocina kr1 was also induced by cis-1,2-dichloroethylene, perchloroethylene, chloroethane ...19957574658
desaturation and oxygenation of 1,2-dihydronaphthalene by toluene and naphthalene dioxygenase.bacterial strains expressing toluene and naphthalene dioxygenase were used to examine the sequence of reactions involved in the oxidation of 1,2-dihydronaphthalene. toluene dioxygenase of pseudomonas putida f39/d oxidizes 1,2-dihydronaphthalene to (+)-cis-(1s,2r)-dihydroxy-1,2,3,4-tetrahydronaphthalene, (+)-(1r)-hydroxy-1,2-dihydronaphthalene, and (+)-cis-(1r,2s)-dihydroxy-1,2-dihydronaphthalene. in contrast, naphthalene dioxygenase of pseudomonas sp. strain ncib 9816/11 oxidizes 1,2-dihydronaph ...19957592326
evidence that operons tcb, tfd, and clc encode maleylacetate reductase, the fourth enzyme of the modified ortho pathway.the maleylacetate reductase from pseudomonas sp. strain b13 functioning in the modified ortho pathway was purified and digested with trypsin. the polypeptides separated by high-performance liquid chromatography were sequenced. alignments with the polypeptides predicted from the tfdf and tcbf genes located on plasmids pjp4 of the 2,4-dichlorophenoxyacetate-degrading alcaligenes eutrophus jmp134 and pp51 of the 1,2,4-trichlorobenzene-degrading pseudomonas sp. strain p51 as well as polypeptides pre ...19957601858
conversion of chlorobiphenyls into phenylhexadienoates and benzoates by the enzymes of the upper pathway for polychlorobiphenyl degradation encoded by the bph locus of pseudomonas sp. strain lb400.metabolism of 21 chlorobiphenyls by the enzymes of the upper biphenyl catabolic pathway encoded by the bph locus of pseudomonas sp. strain lb400 was investigated by using recombinant strains harboring gene cassettes containing bphabc or bphabcd. the enzymes of the upper pathway were generally able to metabolize mono- and dichlorinated biphenyls but only partially transform most trichlorinated congeners investigated: 14 of 15 mono- and dichlorinated and 2 of 6 trichlorinated congeners were conver ...19957618878
isolation and characterization of a pseudomonas sp. that mineralizes the s-triazine herbicide atrazine.a bacterium that was capable of metabolizing atrazine at very high concentrations (>1,000 ppm) was isolated from a herbicide spill site. the organism was differentiated by observing clearing zones on indicator agar plates containing 1,000 ppm atrazine. detailed taxonomic studies identified the organism as a pseudomonas sp., designated adp, that was dissimilar to currently known species. pseudomonas sp. strain adp metabolized atrazine as its sole nitrogen source. nongrowing suspended cells also m ...199516534995
transformation of substituted fluorenes and fluorene analogs by pseudomonas sp. strain f274.pseudomonas sp. strain f274, previously shown to catabolize fluorene via fluorenone and its angular dioxygenation, 2(prm1),3(prm1)-dihydroxy-2-carboxybiphenyl, phthalate, and protocatechuate, was examined for its ability to transform substituted fluorenes and s- and n-heterocyclic analogs. halogen- and methyl-substituted fluorenes were metabolized to correspondingly substituted phthalates via attack on the unsubstituted ring. in the case of 1-methylfluorene, initial oxidation of the methyl group ...199516535132
uptake of benzoic acid and chloro-substituted benzoic acids by alcaligenes denitrificans bri 3010 and bri 6011.the mechanism of uptake of benzoic and 2,4-dichlorobenzoic acid (2,4-dcba) by alcaligenes denitrificans bri 3010 and bri 6011 and pseudomonas sp. strain b13, three organisms capable of degrading various isomers of chlorinated benzoic acids, was investigated. in all three organisms, uptake of benzoic acid was inducible. for benzoic acid uptake into bri 3010, monophasic saturation kinetics with apparent k(infm) and v(infmax) values of 1.4 (mu)m and 3.2 nmol/min/mg of cell dry weight, respectively, ...199516535175
localization and characterization of the carbon tetrachloride transformation activity of pseudomonas sp. strain kc.previous research has established that pseudomonas sp. strain kc rapidly transforms carbon tetrachloride (ct) to carbon dioxide (45 to 55%), a nonvolatile fraction (45 to 55%), and a cell-associated fraction ((equiv)5%) under denitrifying, iron-limited conditions. the present study provides additional characterization of the nonvolatile fraction, demonstrates that electron transfer plays a role in the transformation, and establishes the importance of both extracellular and intracellular factors. ...199516534941
quantification of frankia strains and other root-associated bacteria in pure cultures and in the rhizosphere of axenic seedlings by high-performance liquid chromatography-based muramic acid assay.application of a high-performance liquid chromatography-based muramic acid assay with precolumn fluorescence derivatization to quantification of root-associated bacteria was studied both in pure cultures and in the rhizosphere of axenic festuca rubra seedlings. quantities of muramic acid from acid-hydrolyzed cells of frankia strains, streptomyces griseoviridis, enterobacter agglomerans, klebsiella pneumoniae, pseudomonas sp., and bacillus polymyxa were mostly proportional to the respective cell ...199416349413
insertion sequence is6100 on plasmid poad2, which degrades nylon oligomers.the nucleotide sequence of repeated sequence i, which appears in five regions on nylon oligomer-degrading plasmid poad2, harbored in flavobacterium sp. strain k172, was determined. the five regions of repeated sequence i had 880 bp of identical sequence, and the sequence was identical to that of is6100, an insertion sequence classified in the is6 family, initially found in mycobacterium fortuitum. sequences homologous to that of is6100 were found for another nylon oligomer-degrading plasmid, pna ...19948106333
a biological sensor for iron available to bacteria in their habitats on plant surfaces.a sensor responsive to iron was constructed by fusing a promoterless ice nucleation activity gene (inaz) to an iron-regulated promoter of a genomic region involved in pyoverdine (fluorescent siderophore) (pvd) production in pseudomonas syringae. cells of pseudomonas fluorescens and p. syringae that contained the pvd-inaz fusion expressed iron-responsive ice nucleation activity in the bean rhizosphere and phyllosphere, respectively, and in culture. addition of fe(iii) to leaves or soil reduced th ...199416349283
pseudomonas sp. strain hbp1 prp degrades 2-isopropylphenol (ortho-cumenol) via meta cleavage.pseudomonas sp. strain hbp1 prp grew on 2-isopropylphenol as the sole carbon and energy source with a maximal specific growth rate of 0.14 h-1 and transient accumulation of isobutyric acid. oxygen uptake experiments with resting cells and enzyme assays with crude-cell extracts showed that 2-isopropylphenol was catabolized via a broad-spectrum meta cleavage pathway. these findings were confirmed by experiments with partially purified enzymes. identification of 3-isopropylcatechol and 2-hydroxy-6- ...19947811094
identification and characterization of the treponema pallidum tpn50 gene, an ompa homolog.treponema pallidum is a pathogenic spirochete that has no known genetic exchange mechanisms. in order to identify treponemal genes encoding surface and secreted proteins, we carried out tnphoa mutagenesis of a t. pallidum genomic dna library in escherichia coli. several of the resulting clones expressed enzymatically active t. pallidum-alkaline phosphatase fusion proteins. the dna sequence of the 5' portion of a number of the treponemal genes was obtained and analyzed. a recombinant clone harbor ...19948112835
purification and characterization of thermostable and nonthermostable 2-haloacid dehalogenases with different stereospecificities from pseudomonas sp. strain yl.two novel hydrolytic dehalogenases, thermostable l-2-haloacid dehalogenase (l-dex) inducibly synthesized by 2-chloropropionate (2-cpa) and nonthermostable dl-2-haloacid dehalogenase (dl-dex) induced by 2-chloroacrylate, were purified to homogeneity from pseudomonas sp. strain yl. dl-dex consisted of a monomer with a molecular weight of about 36,000 and catalyzed the dehalogenation of l and d isomers of 2-cpa to produce d- and l-lactates, respectively. it acted on 2-haloalkanoic acids with a carb ...19948074519
comparative studies of genes encoding thermostable l-2-halo acid dehalogenase from pseudomonas sp. strain yl, other dehalogenases, and two related hypothetical proteins from escherichia coli.we have determined the nucleotide sequence of the gene encoding thermostable l-2-halo acid dehalogenase (l-dex) from the 2-chloroacrylate-utilizable bacterium pseudomonas sp. strain yl. the open reading frame consists of 696 nucleotides corresponding to 232 amino acid residues. the protein molecular weight was estimated to be 26,179, which was in good agreement with the subunit molecular weight of the enzyme. the gene was efficiently expressed in the recombinant escherichia coli cells: the amoun ...19947944368
regiospecific and stereoselective hydroxylation of 1-indanone and 2-indanone by naphthalene dioxygenase and toluene dioxygenase.the biotransformation of 1-indanone and 2-indanone to hydroxyindanones was examined with bacterial strains expressing naphthalene dioxygenase (ndo) and toluene dioxygenase (tdo) as well as with purified enzyme components. pseudomonas sp. strain 9816/11 cells, expressing ndo, oxidized 1-indanone to a mixture of 3-hydroxy-1-indanone (91%) and 2-hydroxy-1-indanone (9%). the (r)-3-hydroxy-1-indanone was formed in 62% enantiomeric excess (ee) (r:s, 81:19), while the 2-hydroxy-1-indanone was racemic. ...19947944365
biodegradation of 2-nitrotoluene by pseudomonas sp. strain js42.a strain of pseudomonas sp. was isolated from nitrobenzene-contaminated soil and groundwater on 2-nitrotoluene as the sole source of carbon, energy, and nitrogen. bacterial cells growing on 2-nitrotoluene released nitrite into the growth medium. the isolate also grew on 3-methylcatechol, 4-methylcatechol, and catechol. 2-nitrotoluene, 3-methylcatechol, and catechol stimulated oxygen consumption by intact cells regardless of the growth substrate. crude extracts from the isolate contained catechol ...19947944378
organization and evolution of naphthalene catabolic pathways: sequence of the dna encoding 2-hydroxychromene-2-carboxylate isomerase and trans-o-hydroxybenzylidenepyruvate hydratase-aldolase from the nah7 plasmid.the sequence of a 2,437-bp dna segment from the naphthalene upper catabolic pathway operon of plasmid nah7 was determined. this segment contains three large open reading frames designated nahq', nahe, and nahd. the first of these is the 3' end of an open reading frame that has no known function, the second (993 bp) encodes trans-o-hydroxybenzylidenepyruvate hydratase-aldolase (deduced molecular weight, 36,640), and the third (609 bp) encodes 2-hydroxychromene-2-carboxylate isomerase (deduced mol ...19948002605
oxidative release of nitrite from 2-nitrotoluene by a three-component enzyme system from pseudomonas sp. strain js42.pseudomonas sp. strain js42 utilizes 2-nitrotoluene (2nt) as the sole source of carbon and energy for growth. intact cells catalyze the oxidation of 2nt to 3-methylcatechol and nitrite in a reaction that requires molecular oxygen. cell extracts oxidized 2nt to 3-methylcatechol and nitrite in the presence of nad(p)h and ferrous iron. ion-exchange chromatography yielded three protein fractions (a, b, and c) which were all required for the oxidation of 2nt to 3-methylcatechol and nitrite. component ...19948002568
inability of muconate cycloisomerases to cause dehalogenation during conversion of 2-chloro-cis,cis-muconate.the conversion of 2-chloro-cis,cis-muconate by muconate cycloisomerase from pseudomonas putida prs2000 yielded two products which by nuclear magnetic resonance spectroscopy were identified as 2-chloro- and 5-chloromuconolactone. high-pressure liquid chromatography analyses showed the same compounds to be formed also by muconate cycloisomerases from acinetobacter calcoaceticus adp1 and pseudomonas sp. strain b13. during 2-chloro-cis,cis-muconate turnover by the enzyme from p. putida, 2-chloromuco ...19948021223
cross-regulation by xylr and dmpr activators of pseudomonas putida suggests that transcriptional control of biodegradative operons evolves independently of catabolic genes.the pu promoter of the toluene degradation plasmid pww0 of pseudomonas putida drives expression of an operon involved in the sequential oxidation of toluene and m- and p-xylenes to benzoate and toluates, respectively. similarly, the po promoter of plasmid pvi150 controls expression of an operon of pseudomonas sp. strain cf600 which is required for the complete catabolism of phenol and cresols. these promoters, which both belong to the sigma 54-dependent class, are regulated by their cognate acti ...19948051017
genetic organization of the mau gene cluster in methylobacterium extorquens am1: complete nucleotide sequence and generation and characteristics of mau mutants.the nucleotide sequence of the methylamine utilization (mau) gene region from methylobacterium extorquens am1 was determined. open reading frames for 11 genes (maufbedacjglmn) were found, all transcribed in the same orientation. the maub, maua, and mauc genes encode the periplasmic methylamine dehydrogenase (madh) large and small subunit polypeptides and amicyanin, respectively. the products of maud, maug, maul, and maum were also predicted to be periplasmic. the products of mauf, maue, and maun ...19948021187
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