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presence and quantity of dehydroalanine in histidine ammonia-lyase from pseudomonas putida.dehydroalanine is present in the histidine ammonia-lyase (histidase) from pseudomonas putida atcc 12633 as shown by reaction of purified enzyme with k14cn or nab3h4 and subsequent identification of [14c]aspartate or [3h]alanine, respectively, following acid hydrolysis of the labeled protein. when labeling with cyanide was conducted under denaturing conditions, 4 mol of [14c]cyanide was incorporated per mol of enzyme (mr 220 000), equivalent to one dehydroalanine residue being modified per subuni ...19853919759
measuring the spermosphere colonizing capacity (spermosphere competence) of bacterial inoculants.spermosphere establishment by bacteria which were coated onto seeds was studied using soybean seeds treated with four bacterial strains at levels of log10 1 to 4 colony-forming units (cfu) per seed planted in a field soil mix, and incubated 48 h. each strain at every inoculum level developed spermosphere population densities of log10 4 to 8 cfu/seed, demonstrating an average multiplication of log10 3 cfu/seed. an alternative method was developed to differentially rank bacteria for spermosphere c ...19853933804
[regulation of the synthesis of the key enzymes for naphthalene catabolism in pseudomonas putida and pseudomonas fluorescens carrying the biodegradation plasmids nah, pbs3, pbs2 and npl-1].regulation of the synthesis of key enzymes catalysing naphthalene catabolism was studied in pseudomonas strains containing different plasmids of naphthalene biodegradation. the synthesis of naphthalene oxygenase, salicylate hydroxylase, catechol-1,2-oxygenase and cathechol-2,3-oxygenase was shown to be regulated in both the coordinated and non-coordinated manner.19853937034
regions of broad-host-range plasmid rk2 involved in replication and stable maintenance in nine species of gram-negative bacteria.the replication and maintenance properties of the broad-host-range plasmid rk2 and its derivatives were examined in nine gram-negative bacterial species. two regions of rk2, the origin of replication (oriv) and a segment that encodes for a replication protein (trfa delta kild, designated trfa*), are sufficient for replication in all nine species tested. however, stable maintenance of this minimal replicon (less than 0.3% loss per generation under nonselection conditions) is observed only in esch ...19854044529
impact of cefotaxime on the fecal flora in children.a differential quantitative analysis was used to study the effect of cefotaxime on the fecal flora in 26 hospitalized children ranging from two days to four years of age. fecal specimens were obtained before, during and after therapy. this study was evaluated in comparison to 41 patients of the same age and from the same environment without antibiotic treatment or signs of infection. the fecal flora of the control group showed qualitative and quantitative stability. two groups of species were di ...19854055046
dehalogenase genes of pseudomonas putida pp3 on chromosomally located transposable elements.pseudomonas putida pp3 utilizes halogenated alkanoic acids (haa) such as 2,2-dcpa as its sole carbon and energy sources. spontaneous hha- mutants, isolated by selection for resistance to the toxic analogs monochloroacetic acid and dichloroacetic acid, arose at frequencies several orders of magnitude higher than expected for spontaneous mutations. analysis of the five classes of mutants isolated suggested that the dehalogenase and haa permease genes were on chromosomally located transposable elem ...19852835577
[expression of pseudomonas aeruginosa transposable phages in pseudomonas putida cells. i. the establishment of a lysogenic state and the effectiveness of lytic development].expression of transposable phages (tp) of pseudomonas aeruginosa in the cells of p. putida was studied. the high efficiency of phage lytic development was shown both as a consequence of zygotic induction after transfer of the rp4::tpc+ plasmid into nonlysogenic recipients, and as a result of heat induction of lysogens ppg1 (d3112cts15). the high phage yield (20-25 particles of d3112cts phage per one cell of p. putida) is an evidence for a high level of transposition in the cells of this bacteria ...19852998927
[expression of pseudomonas aeruginosa phage transposons in pseudomonas putida ppg1 cells. ii. zygotic induction--a necessary condition for the formation of defective lysogens].the transfer of hybrid plasmid rp4::pt (where pt is the genome of a transposable phage specific for pseudomonas aeruginosa) into recipient cells of p. putida strain ppg1 occurs with the same frequency as into p. aeruginosa, the homologous host for pt. approximately 1/3 of all ppg1 exconjugants carrying rp4 markers lost the capability to produce viable pt phage. in contrast, in a cross with homologous recipient p. aeruginosa all exconjugant clones contained nondefective prophages in the hybrid pl ...19853002910
[hybrid plasmid pbs251 containing genes for n-alkane degradation].the strain of pseudomonas aeruginosa bs316 utilizing h-alkanes of the c6-c12 series (alk+) harbours the 96 md plasmid pbs250. the use of plasmid rp4 to mobilize alk+ markers in conjugal transfer to pseudomonas aeruginosa and pseudomonas putida has resulted in isolation of transconjugants resistant to antibiotics (due to genes coded by plasmid rp4) and capable of growth on h-alkanes. a transconjugant from this series harbours plasmid pbs251, a derivative of plasmid rp4 containing the genes for oc ...19853025683
[17o]water and nitric oxide binding by protocatechuate 4,5-dioxygenase and catechol 2,3-dioxygenase. evidence for binding of exogenous ligands to the active site fe2+ of extradiol dioxygenases.pseudomonas testosteroni protocatechuate 4,5-dioxygenase and pseudomonas putida catechol 2,3-dioxygenase (metapyrocatechase) catalyze extradiol-type oxygenolytic cleavage of the aromatic ring of their substrates. the essential active site fe2+ of each enzyme binds nitric oxide (no) to produce an epr active complex with an electronic spin of s = 3/2. hyperfine broadening of the epr resonances of the nitrosyl complexes by 17o-enriched h2o shows that water is bound directly to the fe2+ in the nativ ...19852997190
cloning of the gene for the common pathogenic neisseria h.8 antigen from neisseria gonorrhoeae.neisseria gonorrhoeae dna that encodes the pathogenic neisseria h.8 common antigen was cloned in the lambda phage sep6. the recombinant phage, designated s6h.8, was detected immunologically with a monoclonal antibody that binds to the h.8 antigen. the gonococcal and s6h.8 forms of the antigen yielded identical partial proteolysis epitope maps. neisseria species that did not manifest the h.8 antigen showed little or no dna homology with s6h.8. this clone should facilitate investigation into the c ...19852981198
purification and properties of ferredoxintol. a component of toluene dioxygenase from pseudomonas putida f1.toluene dioxygenase oxidizes toluene to (+)-cis-1(s),2(r)-dihydroxy-3-methylcyclohexa-3,5-diene. this reaction is catalyzed by a multienzyme system that is induced in cells of pseudomonas putida f1 during growth on toluene. one of the components of toluene dioxygenase has been purified to homogeneity and shown to be an iron-sulfur protein that has been designated ferredoxintol. the molecular weight of ferredoxintol was calculated to be 15,300, and the purified protein was shown to contain 2 g of ...19852982815
p450cam gene cloning and expression in pseudomonas putida and escherichia coli.the gene camc, which encodes the cytochrome p450 monoxygenase protein, was cloned into the shuttle vector pkt240 and recovered as the recombinant pkg201 with a 2.3 kb insert from the cam plasmid in the psti site. the gene product is expressed constitutively in p. putida and in e. coli whereas the inverted insert clone lacks expression, indicating absence of an insert promoter.19852992468
determination of the transcription initiation site and identification of the protein product of the regulatory gene xylr for xyl operons on the tol plasmid.the xylr gene is a regulatory gene on the tol plasmid, which acts in a positive manner on xyl operons for degradation of toluene and xylenes in pseudomonas putida. a dna fragment containing the xylr promoter region was cloned on promoter-probing vectors, and its nucleotide sequence was determined. the transcription initiation site of the xylr gene was determined in cells of p. putida and escherichia coli by s1 nuclease and reverse transcriptase mapping. two initiation sites were detected which w ...19852993247
evolutionary conservation of genes coding for meta pathway enzymes within tol plasmids pww0 and pww53.pseudomonas putida mt53 contains a tol plasmid, pww53, that encodes toluene-xylene catabolism. pww53 is nonconjugative, is about 105 to 110 kilobase pairs (kbp) in size, and differs significantly in its restriction endonuclease digestion pattern and incompatibility group from the archetypal tol plasmid pww0. an rp4::pww53 cointegrate plasmid, pww53-4, containing about 35 kbp of pww53 dna, including the entire catabolic pathway genes, was formed, and a restriction map for kpni, hindiii, and bamhi ...19852997136
omega mutagenesis in gram-negative bacteria: a selectable interposon which is strongly polar in a wide range of bacterial species.we have used the 2.0-kb dna fragment omega [prentki and krisch, gene 29 (1984) 303-313] to mutagenize in vitro a broad-host-range plasmid carrying the entire meta-cleavage pathway of the pseudomonas putida tol plasmid pww0. the mutant plasmids were subsequently introduced by conjugal mobilization into a variety of gram-negative bacteria. the omega fragment carries a selectable marker (aada+; spcr/smr), which is expressed in all species tested, as well as flanking transcription and translation te ...19852998930
cloning and expression in escherichia coli of histidine utilization genes from pseudomonas putida.a library of the pseudomonas putida chromosome, prepared through the use of the cosmid pjb8 ligated to a partial sau3a digest of bacterial dna, followed by in vitro packaging into bacteriophage lambda particles, was used to construct a strain of escherichia coli which contained the genes for histidine utilization. this isolate produced a repressor product and all five enzymes required in pseudomonas spp. for histidine dissimilation, whereas none of these could be detected in the nontransduced pa ...19852858467
in vivo role of sulfite in photocontrol of urocanase from pseudomonas putida. 19852858892
sal-tol in vivo recombinant plasmid pkf439.sal-tol in vivo recombinant plasmid pkf439 was characterized in a strain from a mixed culture of bacteria harboring various degradative plasmids. analysis of the gene organization of pkf439 revealed that the 57-kilobase tol fragment, including the 40-kilobase tol metabolic region, was inserted into the complete sal replicon at the position of smai-c within xhoi-b of sal. the molecular size of pkf439 was calculated to be 138 kilobases. pkf439 could be transferred to pseudomonas putida and pseudom ...19852987190
shuttle vector for escherichia coli, pseudomonas putida, and pseudomonas aeruginosa.a hybrid plasmid capable of replication in 2 different genera, escherichia and pseudomonas, was constructed. this plasmid dna can be used as a cloning vector in e. coli and pseudomonades cells. the described hybrid plasmid pld411 has been constructed on the basis of 2 small e. coli vector r-plasmids used in our laboratory and cryptic plasmid pww2 or p. putida mt1. plasmid pld411 dna was mapped with restrictases; its biological activity in transformations of different bacterial strains was studie ...19852990432
p-cresol methylhydroxylase. assay and general properties.p-cresol methylhydroxylase from pseudomonas putida, an anaerobic dehydrogenase that catalyses the oxidation of p-cresol to p-hydroxybenzyl alcohol and then to p-hydroxybenzaldehyde, is an enzyme of great interest in several respects. one of these is the fact that its flavoprotein and cytochrome c subunits may be reversibly dissociated with ease, with full regeneration of the activity and its native properties on recombining the components. bisubstrate kinetic analysis of the unresolved enzyme gi ...19852990444
activation of urocanase from pseudomonas putida by electronically excited triplet species.urocanase from pseudomonas putida becomes inactive in growing and resting cells and, as shown previously, is activated by the direct absorption of ultraviolet light. in this study, we describe the activation of urocanase by energy transfer from triplet indole-3-aldehyde, generated in the peroxidase-catalyzed aerobic oxidation of indole-3-acetic acid. the activation was time-, temperature-, and ph-dependent. the involvement of reactive oxygen intermediates was excluded by the lack of effect of ap ...19852864338
isolation and analysis of genes involved in siderophore biosynthesis in plant-growth-stimulating pseudomonas putida wcs358.the plant-growth-stimulating pseudomonas putida wcs358 was mutagenized with transposon tn5. the resulting mutant colony bank was screened for mutants defective in the biosynthesis of the fluorescent siderophore. a total of 28 mutants, divided into six different classes, were isolated that were nonfluorescent or defective in iron acquisition or both. these different types of mutants together with the probable overall structure of the siderophore, i.e., a small peptide chain attached to a fluoresc ...19852997118
tryptophanyl fluorescence quenching of urocanase from pseudomonas putida by acrylamide, cesium, iodide, and imidazolepropionate. 19852864711
expression of plasmid encoded escherichia coli 5s ribosomal ribonucleic acid in pseudomonas putida.the recombinant plasmid pnrk 36, which represents the plasmids rsf 1010, a small multicopy plasmid of the incompatibility group incq that confers resistance to streptomycin and sulfoamide to its host cells, and pkk 223-3, which contains the try-lac (tac) promoter followed by a polylinker and a dna segment containing the 5s rrna (rrn b) with the ribosomal rna transcription terminators, was employed to transform pseudomonas putida 2440 cells. the plasmid encoded 5 s rrna from escherichia coli was ...19852411598
the kinetics of dihydrostreptomycin uptake in pseudomonas putida membrane vesicles: absence of inhibition by cations.dihydrostreptomycin was taken up in isolated cytoplasmic membrane vesicles of pseudomonas putida by an active transport mechanism. saturation kinetics were observed with an apparent km and vmax of 15 mm and 50 nmol/min/mg of protein respectively. the evidence suggested that the observed kinetics was that of the energy-dependent phase i component of dihydrostreptomycin uptake. neither magnesium nor the polyamine, spermine, inhibited dihydrostreptomycin transport. thus, the inhibition of aminoglyc ...19852415504
characterization of two surface-localized antigenic sites on porin protein f of pseudomonas aeruginosa.a rapid colony immunoblot screening procedure was used to demonstrate the surface localization of porin protein f on bacterial colonies of pseudomonas aeruginosa. by this method, we demonstrated that protein f was accessible to four different specific monoclonal antibodies in a wide variety of both mucoid and nonmucoid p. aeruginosa strains. controls were performed to demonstrate that, using this procedure, only surface-exposed epitopes bound monoclonal antibodies and that nonspecific binding of ...19852408719
homology between nucleotide sequences of promoter regions of nah and sal operons of nah7 plasmid of pseudomonas putida.the in vivo transcription start sites of the nah and sal operons of the nah7 plasmid were determined by s1 nuclease mapping and the nucleotide sequence surrounding these transcription start sites was determined. since expression of both of these operons is coordinately controlled by the product of the transcriptional activator gene nahr, the sequences were compared to locate potential sites involved in common regulation. in the 100-base-pair region preceding transcription start sites of both ope ...19863001734
characterization of a plasmid-specified pathway for catabolism of isopropylbenzene in pseudomonas putida re204.a pseudomonas putida strain designated re204, able to utilize isopropylbenzene as the sole carbon and energy source, was isolated. tn5 transposon mutagenesis by means of the suicide transposon donor plasmid plg221 yielded mutant derivatives defective in isopropylbenzene metabolism. these were characterized by the identification of the products which they accumulated when grown in the presence of isopropylbenzene and by the assay of enzyme activities in cell extracts. based on the results obtaine ...19863019995
characterization of the oct plasmid encoding alkane oxidation and mercury resistance in pseudomonas putida.transformation of pseudomonas putida and analysis for plasmid dna revealed that both n-alkane oxidation and mercury resistance are encoded on a single 220-megadalton oct plasmid molecule. derivatives of oct having lost the mercury resistance function could be readily isolated and contained a smaller plasmid estimated to be 170 megadaltons. the results show that segregation of the mercury resistance property occurs not by loss of a separate mer plasmid as previously thought but by a deletion in t ...19863003035
nucleotide sequence of the pseudomonas putida cytochrome p-450cam gene and its expression in escherichia coli.cytochrome p-450cam catalyzes the stereospecific methylene hydroxylation of camphor to form 5-exohydroxycamphor and is encoded by the camc gene on the cam plasmid of pseudomonas putida, atcc 17453. the cytochrome p-450cam structural gene has been cloned by mutant complementation in p. putida (koga, h., rauchfuss, b., and gunsalus, i. c. (1985) biochem. biophys. res. commun. 130, 412-417). we report the complete nucleotide sequence of the camc gene along with 155 base pairs of 5' and 175 base pai ...19863003058
analysis of the vegetative replication origin of broad-host-range plasmid rk2 by transposon mutagenesis.a range of tn1723 transposon mutants of the oriv region of broad-host-range plasmid rk2 have been isolated, and the internal ecori fragment of the transposon has been deleted from each to reduce the insertion size from 9.6 kb (tn1723) to 35 bp (delta tn1723). sequencing from the delta tn1723-derived ecori site has allowed the precise mapping of these insertions to various points dispersed through the origin region. using these mutants we have determined which regions of oriv rk2 are of functiona ...19863010353
utilization of pyrimidines and pyrimidine analogues by fluorescent pseudomonads.the fluorescent pseudomonads pseudomonas aeruginosa, pseudomonas aureofaciens and pseudomonas putida were examined for their ability to utilize pyrimidines and pyrimidine analogues. both p. aeruginosa and p. aureofaciens grew upon dihydrouracil, dihydrothymine, uridine and cytidine as either a sole nitrogen or carbon source while uracil, thymine and cytosine served only as sole nitrogen sources for both pseudomonads. the only difference between the observed growth of p. aeruginosa and p. aureofa ...19863097460
a comparative study of acquired amidase activity in pseudomonas species.pseudomonas putida pp3 carrying dehalogenases i and ii and pseudomonas aeruginosa pau3 carrying dehalogenase i coded for by plasmid puu2 were able to grow on 2-monochloropropionic acid (2mcpa). neither strain utilized 2-chloropropionamide (2cpa) as a carbon or nitrogen source for growth. mutations in both strains to 2cpa+ phenotypes (designated p. putida ppw3 and p. aeruginosa pau5, respectively) involved the expression of an acquired 2cpa-amidase activity. the amidase followed by dehalogenase r ...19863098906
camr, a negative regulator locus of the cytochrome p-450cam hydroxylase operon.a 4.27-kilobase insert from a hindiii dna library of pseudomonas putida carrying the cam plasmid allowed coordinate expression of genes camd and camc under control of camr, an upstream regulator. the camc gene specifies cytochrome p-450cam, and camd specifies the 5-exo-alcohol dehydrogenase. a 1.38-kilobase deletion from the insert results in the constitutive expression of genes camc and camd; transformation in trans restores the substrate control, indicating that camr is a negative regulator.19863011733
l-malate transport and proton symport in vesicles prepared from pseudomonas putida.in vesicles from glucose-grown pseudomonas putida, l-malate is transported by nonspecific physical diffusion. l-malate also acts as an electron donor and generates a proton motive force (delta p) of 129 mv which is composed of a membrane potential (delta psi) of 60 mv and a delta ph of 69 mv. in contrast, vesicles from succinate-grown cells transport l-malate by a carrier-mediated system with a km value of 14.3 mm and a vmax of 313 nmol x mg protein-1 x min-1, generate no delta psi, delta ph, or ...19863030368
[comparative analysis of the organization of the npl-1 plasmid controlling naphthalene oxidation in pseudomonas putida and its derivatives].the paper contains the data on the structure of npl-1 plasmid controlling naphthalene biodegradation. the plasmid which pertains to the p-9 incompatibility group is transferrable conjugatively and is maintained stably within a wide range of gram-negative bacteria. the analysis of mutants and transposon derivatives of the plasmid made it possible to localize nah-genes in a dna fragment, 23 kb in size. an inverted dna segment of 4.2 kb was discovered which may participate in the regulation of nah- ...19863025060
[nah-genes of pseudomonas putida: molecular genetic analysis of the plasmid pbs286].the hybridization and restriction analysis of the plasmid pbs286 (73 kb, the p-9 inc group) as well as parental plasmids npl-1, npl-41 demonstrated that pbs286 plasmid (delta npl-41::tna) with the constitutive synthesis of naphthalene dioxygenase carried genes for naphthalene oxidation to salicylate and those participating in degradation of catechol. restriction map of pbs286 using xhoi restriction endonuclease and that of the nah region using ecori, bamhi, sali and xhoi were established. struct ...19863026897
[similar organization of beta,beta'-subunit rna-polymerase genes and adjacent ribosomal protein genes in enterobacteriaceae and pseudomonas putida].the genes coding for the rna-polymerase beta,beta'-subunits and adjacent ribosomal protein genes in escherichia coli, salmonella typhimurium, shigella flexneri, serratia marcescens, proteus mirabilis and pseudomonas putida are compared by the southern hybridization procedure. in all the species studied close clustering of the genes rplkajl and rpobc is demonstrated. preliminary physical maps for these genes in s. typhimurium, s. flexneri, s. marcescens and p. mirabilis are proposed. rifampicin i ...19863005845
plasmid-determined cadmium resistance in pseudomonas putida gam-1 isolated from soil.cadmium-resistant pseudomonas putida gam-1, which was able to grow in concentrations of cdcl2 as high as 7 mm, was isolated from soil in a rice paddy. this bacterium harbored a dna plasmid of about 52 kilobases. the plasmid (pgu100) transformed escherichia coli c600 to cadmium resistance. a cadmium-resistant transformant of e. coli c600 contained a plasmid corresponding to that seen in p. putida gam-1. the transformant did not take up cadmium as well as p. putida gam-1 did.19863941050
comparative analysis of different pseudomonas strains that degrade cinnamic acid.strains of pseudomonas stutzeri (cinns) and pseudomonas putida (cinnp and cinnw) isolated from soil with cinnamic acid as the sole carbon source were found to be simultaneously adapted to grow on phenylpropionic and p-hydroxybenzoic acids. in cinnamic acid-grown cultures, phenylpropionic acid was isolated. a catabolic plasmid of approximately equal to 75 kilobase pairs encoding the metabolism of cinnamic acid was found in strains cinnp and cinns.19863777934
thermodynamic properties of oxidation-reduction reactions of bacterial, microsomal, and mitochondrial cytochromes p-450: an entropy-enthalpy compensation effect.an optically transparent thin-layer electrode cell with a very small volume was used for determination of the formal reduction potentials of bacterial, microsomal, and mitochondrial cytochromes p-450. at an extrapolated zero concentration of dye, the bacterial cytochrome from pseudomonas putida catalyzing the hydroxylation of camphor and the adrenal mitochondrial cytochrome catalyzing the cholesterol side-chain cleavage reaction had formal reduction potentials of -168 and -285 mv (ph 7.5 and 25 ...19863964682
primary alkylsulphatase activities of the detergent-degrading bacterium pseudomonas c12b. purification and properties of the p1 enzyme.the p1 primary alkylsulphatase of pseudomonas c12b was purified 1500-fold to homogeneity by a combination of streptomycin sulphate precipitation of nucleic acids, (nh4)2so4 fractionation and chromatography on columns of deae-cellulose, sephacryl s-300 and butyl-agarose. the protein was tetrameric with an mr of 181000-193000, and exhibited maximum activity at ph 6.1. primary alkyl sulphates of carbon-chain length c1-c5 or above c14 were not substrates, but the intermediate homologues were shown t ...19863753455
chromosomal location of tol plasmid dna in pseudomonas putida.the soil isolate pseudomonas putida mw1000 can grow on toluene and other hydrocarbons; in this respect it is similar to strains of pseudomonas which carry the tol plasmid. by conjugation experiments, the genes conferring these growth abilities have been shown to be located on the bacterial chromosome, linked to vil and catb. a 56-kilobase segment of the bacterial chromosome of mw strains carrying the tol genes can transpose to the incp-1 plasmid r18-18. physical analysis of these tol r18-18 hybr ...19863782038
amino acid and sequence analysis of the cytochrome and flavoprotein subunits of p-cresol methylhydroxylase.the flavocytochrome p-cresol methylhydroxylase from pseudomonas putida has been reported to have a mr of 114,000 and to consist of two subunits, a flavoprotein and a cytochrome c, each with a mr of 58,000. recent x-ray crystallographic data from our laboratories [shamala, n., lim, l. w., mathews, f. s., mcintire, w., singer, t. p., & hopper, d. j. (1986) proc. natl. acad. sci. u.s.a. 83, 4626-4630], however, indicate an alpha 2 beta 2 structure and a much lower molecular mass (approximately 8000 ...19863790500
naphthalene association and uptake in pseudomonas putida.two methods for bacterial membrane transport, filtration and flow dialysis, were used to study cellular association of pseudomonas putida with naphthalene. it is not technically possible to determine the exact cellular or vesicular location of the naphthalene, and because it is hydrophobic, it could be at the membrane(s) rather than inside the cells. as an index of naphthalene having crossed the inner membrane we used the intracellular formation of its first catabolite. an energized membrane or ...19863957866
the amino acid sequence of a delta 5-3-oxosteroid isomerase from pseudomonas putida biotype b.we have determined the primary structure of a delta 5-3-oxosteroid isomerase from pseudomonas putida biotype b. the enzyme is a dimeric protein of two identical subunits, each consisting of a polypeptide chain of 131 residues and a mr = 14,536. the intact s-carboxymethyl protein was sequenced from the nh2 terminus using standard automated edman degradation and automated edman degradation using fluorescamine treatment at known prolines to suppress background. the isomerase was fragmented using cn ...19863700400
cadmium-binding proteins in pseudomonas putida: pseudothioneins.pseudomonas putida adapted to growth in 3 mm cadmium. the resistance mechanism involved complexation of cadmium in polyphosphate granules, changes in the structure of the cell membrane and induction of three cysteine-rich, low molecular weight proteins (3500-7000) containing 4 to 7 g-atoms per mole of cadmium, zinc, and copper. each protein was produced during a different phase of growth, and the smallest protein (3500) was released into the environment when the cells lysed at the end of the exp ...19863709466
identification of cis-diols as intermediates in the oxidation of aromatic acids by a strain of pseudomonas putida that contains a tol plasmid.pseudomonas putida bg1 was isolated from soil by enrichment with p-toluate and selection for growth with p-xylene. other hydrocarbons that served as growth substrates were toluene, m-xylene, 3-ethyltoluene, and 1,2,4-trimethylbenzene. the enzymes responsible for growth on these substrates are encoded by a large plasmid with properties similar to those of tol plasmids isolated from other strains of pseudomonas. treatment of p. putida bg1 with nitrosoguanidine led to the isolation of a mutant stra ...19863711022
influence of para-substituents on the oxidative metabolism of o-nitrophenols by pseudomonas putida b2.pseudomonas putida b2 is able to grow on o-nitrophenol (onp) as the sole source of carbon and nitrogen. onp was converted by a nitrophenol oxygenase to nitrite and catechol. catechol was then attacked by a catechol 1,2-dioxygenase and further degraded through an ortho-cleavage pathway. onp derivatives which were para-substituted with a methyl-, chloro-, carboxy-, formyl- or nitro-group failed to support growth of strain b2. relevant catabolic enzymes were characterized to analyze why these deriv ...19863752997
cloning of pseudomonas sp. strain cbs3 genes specifying dehalogenation of 4-chlorobenzoate.the degradation of 4-chlorobenzoate (4-cba) by pseudomonas sp. strain cbs3 is thought to proceed first by the dehalogenation of 4-cba to 4-hydroxybenzoate (4-hba), which is then metabolized following the protocatechuate branch of the beta-ketoadipate pathway. the cloning of the 4-cba dehalogenation system was carried out by constructing a gene bank of pseudomonas sp. strain cbs3 in pseudomonas putida. hybrid plasmid ppsa843 contains a 9.5-kilobase-pair fragment derived from the chromosome of pse ...19863759912
crystal structure of substrate-free pseudomonas putida cytochrome p-450.the crystal structure of pseudomonas putida cytochrome p-450cam in the substrate-free form has been refined at 2.20-a resolution and compared to the substrate-bound form of the enzyme. in the absence of the substrate camphor, the p-450cam heme iron atom is hexacoordinate with the sulfur atom of cys-357 providing one axial heme ligand and a water molecule or hydroxide ion providing the other axial ligand. a network of hydrogen-bonded solvent molecules occupies the substrate pocket in addition to ...19863768350
a catecholic 9,10-seco steroid as a product of aerobic catabolism of cholic acid by a pseudomonas sp.a mutant of the efficient bile acid-utilizing pseudomonas putida atcc 31752 was found to accumulate three major catabolites on aerobic growth on cholic acid. one of these catabolites was isolated and identified as 3,4,7,12 beta-tetrahydroxy-9,10-seco-1,3,5(10)-androstatriene-9,17-dione (2). this is the first catecholic 9,10-secosteroid isolated from the microbial degradation of bile acids or sterols and confirms the role of such secosteroids in the microbial degradative pathway for steroids.19863445293
formaldehyde dismutase, a novel nad-binding oxidoreductase from pseudomonas putida f61.a novel enzyme, formaldehyde dismutase, was purified and crystallized from the cell extract of an isolated bacterium, pseudomonas putida f61. the enzyme catalyzes the dismutation of aldehydes and alcohol:aldehyde oxidoreduction in the absence of an exogenous electron acceptor. the enzyme is composed of four identical subunits with a mr of 44 000. each subunit contains 1 mol nad(h) and 2 mol zinc/mol. the ratio of nad+ and nadh in a crystalline preparation of the enzyme was about 7:3. the enzyme- ...19863514215
[in vitro bacteriostatic activity of piperacillin sodium against pseudomonas].one hundred and twenty-two strains of pseudomonas isolated from diverse pathological processes were typified. in vitro activity of piperacillin (antibiogram and mic) was studied and compared with another two semisynthetic penicillins, carbenicillin and mezlocillin, and two aminoglycosides, amikacin and gentamicin. the strains isolated corresponded to: pseudomonas aeruginosa (116), pseudomonas cepacia (3), pseudomonas fluorescens (1) and pseudomonas putida (1). it was found that 110 pseudomonas a ...19863685380
leads from the mmwr. reported contamination of heparin sodium with pseudomonas putida. 19863517390
[catabolic pathway of biphenyl controlled by the plasmid pbs241 in pseudomonas putida bs893]. 19863720511
[selection of escherichia coli and pseudomonas putida cultures with an enhanced resistance to immobilization on polyacrylamide gel].clones of escherichia coli (a4, a70, g60) and pseudomonas putida (a70, g30) with an elevated resistance to the process of immobilization in polyacrylamide gel and to the action of monomeric acrylamide were selected from the parent e. coli ibpm b115 and p. putida. the isolated cultures remained resistant to the above actions for a long time. the frequency at which cells with the elevated resistance appeared was comparable with the frequency of bacterial mutations. the plasmid analysis did not rev ...19863517599
the xylabc promoter from the pseudomonas putida tol plasmid is activated by nitrogen regulatory genes in escherichia coli.the xylabc promoter (op1), located on the tol plasmid of pseudomonas putida contains sequences homologous to the conserved regions found in nitrogen fixation (nif) promoters and in other promoters subject to nitrogen control. xyla-lac fusions were constructed in order to monitor expression from the op1 promoter in escherichia coli. transcription was activated in the presence of the heterologous regulatory genes ntrc or nifa from klebsiella pneumoniae as well as by the homologous p. putida regula ...19863520241
vector for regulated expression of cloned genes in a wide range of gram-negative bacteria.a pkt231-based broad-host-range plasmid vector was constructed which enabled regulation of expression of cloned genes in a wide range of gram-negative bacteria. this vector, pnm185, contained upstream of its ecori, ssti, and sstii cloning sites the positively activated pm twin promoters of the tol plasmid and xyls, the gene of the positive regulator of these promoters. expression of cloned genes was induced with micromolar quantities of benzoate or m-toluate, the inexpensive coinducers of the pm ...19863525513
genetic analysis of a relaxed substrate specificity aromatic ring dioxygenase, toluate 1,2-dioxygenase, encoded by tol plasmid pww0 of pseudomonas putida.toluate 1,2-dioxygenase is the first enzyme of a meta-cleavage pathway for the oxidative catabolism of benzoate and substituted benzoates to krebs cycle intermediates that is specified by tol plasmid pww0 of pseudomonas putida. a collection of derivatives harbouring tn1000 insertions and defective in toluate dioxygenase have been isolated from ppl392, a pbr322-based hybrid plasmid carrying the tol plasmid meta-cleavage pathway operon. in parallel, a series of n-methyl-n'-nitro-n-nitro-soguanidin ...19863010045
purification and characterization of glyoxalase i from pseudomonas putida.glyoxalase i was purified to apparent homogeneity from pseudomonas putida. the enzyme was a monomer with a molecular weight of 20,000. the enzyme was most active at ph 8.0. the km values for methylglyoxal and 4,5-dioxovale-rate are 3.5 mm and 1.2 mm, respectively. contrary to the case of eukaryotic enzymes, chelating agents showed little inhibitory effects on the enzyme activity. among the metal ions tested, zn++ specifically and completely inhibited the activity of the enzyme at a millimolar le ...19863101683
in vitro activity of imipenem towards gram-negative bacilli.the authors studied the in vitro antimicrobial activity of imipenem towards 355 gram-negative bacterial strains, taking into particular consideration unusual or dangerous species. the study was carried out on a comparative basis with piperacillin, cefotaxime, ceftazidime and gentamicin. ninety percent of the fermentative gram-negative strains were inhibited at concentrations less than or equal to 2 mg/l. imipenem inhibited 100% of strains of alcaligenes faecalis, alcaligenes denitrificans, flavo ...19863466723
identification of legionella pneumophila with commercially available immunofluorescence test. 19863522642
reactions of 3-ethylcatechol and 3-(methylthio)catechol with catechol dioxygenases.the reactions of 3-ethylcatechol and 3-(methylthio)catechol with catechol 1,2-dioxygenase and catechol 2,3-dioxygenase from pseudomonas putida were examined. both 3-substituted catechols are oxidized by catechol 2,3-dioxygenase at approximately 30% of the rate observed for catechol oxidation by this enzyme. analysis of the products of the reactions showed that ring cleavage occurs in a normal fashion between carbons 2 and 3 of the alternate substrates. 3-ethylcatechol is oxidized by catechol 1,2 ...19863015028
cloning and expression of acinetobacter calcoaceticus catbcde genes in pseudomonas putida and escherichia coli.this report describes the isolation and preliminary characterization of a 5.0-kilobase-pair (kbp) ecori dna restriction fragment carrying the catbcde genes from acinetobacter calcoaceticus. the respective genes encode enzymes that catalyze four consecutive reactions in the catechol branch of the beta-ketoadipate pathway: catb, muconate lactonizing enzyme (ec 5.5.1.1); catc, muconolactone isomerase (ec 5.3.3.4); catd, beta-ketoadipate enol-lactone hydrolase (ec 3.1.1.24); and cate, beta-ketoadipa ...19863003031
spontaneous deletion of a 20-kilobase dna segment carrying genes specifying isopropylbenzene metabolism in pseudomonas putida re204.the genes encoding isopropylbenzene metabolism in pseudomonas putida re204 are readily lost in two ways: by loss (curing) of plasmid pre4 which specifies the catabolic pathway and by deletion from pre4 of an approximately 20-kilobase segment of dna carrying the catabolic genes. the presence of dna sequences at the ends of the catabolic gene region sharing homology with one another suggests that the deletions result from recombination events between these homologous sequences.19863020004
nucleotide sequence of a dna segment promoting transcription in pseudomonas putida.a dna segment that promotes gene expression in pseudomonas putida was identified in ptn8, a mutant plasmid of an rp4-tol recombinant. a promoter on the segment was cloned with a promoter-probe vector containing the xyle gene of the tol plasmid. the xyle gene was expressed under the control of the promoter, and the gene product catechol 2,3-dioxygenase was constitutively synthesized. as analyzed by an s1 nuclease protection assay, the amount of mrna produced in p. putida was more than that in esc ...19863011741
nucleotide sequence of the regulatory gene xyls on the pseudomonas putida tol plasmid and identification of the protein product.the xyls gene is a regulatory gene which positively controls expression of the genes on the tol plasmid for degradation enzymes of benzoate or m-toluate in pseudomonas putida. cloning of the gene in escherichia coli and determination of the nucleotide sequence revealed an open reading frame of 963 bp which corresponds to a protein with an mr of 36,502. the xyls gene was recloned onto a tac-promoter vector, and the product was identified by the maxicell procedure as a protein with an approximate ...19863023186
reported contamination of heparin sodium with pseudomonas putida. 19863080667
amino acid sequence of the pyruvate and the glyoxylate active-site lysine peptide of escherichia coli 2-keto-4-hydroxyglutarate aldolase.pure 2-keto-4-hydroxyglutarate aldolase of escherichia coli, a "lysine-type" trimeric enzyme which has the unique properties of forming an "abortive" schiff-base intermediate with glyoxylate (the aldehydic product/substrate) and of showing strong beta-decarboxylase activity toward oxalacetate, binds any one of its substrates (2-keto-4-hydroxyglutarate, pyruvate, or glyoxylate) in a competitive manner. to determine whether the substrates bind at the same or different (juxta-positioned) sites and ...19863090043
adaptation of pseudomonas putida mt-2 to growth on aromatic amines.pseudomonas putida mt-2 (atcc 33015) carrying the tol plasmid pww0 could adapt to growth on the aromatic amines aniline and m- and p-toluidine. in strain ucc2, a derivative adapted to rapid growth on these compounds, they were oxidatively deaminated to catechol or 4-methylcatechol, which in turn were dissimilated by a meta-cleavage pathway. the aniline/toluidine oxygenase and the meta-cleavage pathway enzymes were inducible by aromatic amines. evidence is presented that in strain ucc2, plasmid p ...19863794647
camphor revisited: studies of 2,5-diketocamphane 1,2-monooxygenase from pseudomonas putida atcc 17453.the oxygenating component of 2,5-diketocamphane 1,2-monooxygenase from pseudomonas putida atcc 17453 was purified to homogeneity by a combination of ammonium sulfate fractionation and chromatography on deae-cellulose and polyanion si-17 columns. it had an mr of 78,000, bound one molecule of nonautooxidizable flavin mononucleotide (fmn), consisted of two subunits of equal molecular weight, and existed in two electrophoretically distinguishable active forms. the oxygenating complex was constructed ...19863944058
purification and characterization of a novel enzyme, n-carbamoylsarcosine amidohydrolase, from pseudomonas putida 77.n-carbamoylsarcosine amidohydrolase, a novel enzyme involved in the microbial degradation of creatinine in pseudomonas putida 77, was purified 27-fold to homogeneity with a 63% overall recovery through simple purification procedures including successive ammonium sulfate fractionation, deae-cellulose chromatography, and crystallization. the relative molecular mass of the native enzyme estimated by the ultracentrifugal equilibrium method is 102,000 +/- 5000, and the subunit mr is 27,000. the km an ...19863745168
bioaccumulation of germanium by pseudomonas putida in the presence of two selected substrates.the uptake of germanium by pseudomonas putida atcc 33015 was studied in the presence of catechol or acetate or both as representative substrates differing in their ability to form complexes with this element. the bacteria were taken from a batch culture grown on acetate as the sole carbon source. cells introduced into a medium containing germanium and either catechol or a mixture of catechol and acetate accumulated germanium in a biphasic way. after a lower level of accumulation that corresponde ...198616347056
binding of germanium to pseudomonas putida cells.the binding of germanium to pseudomonas putida atcc 33015 was investigated by using whole intact cells grown in a medium supplemented with geo(2) and catechol or acetate. electron-microscopic examination of the control and metal-loaded samples revealed that germanium was bound within the cell envelope. a certain number of small electron-dense deposits of the bound element were found in the cytoplasm when the cells were grown in the presence of geo(2) and catechol. the study of germanium distribu ...198616347062
toluene induction and uptake kinetics and their inclusion in the specific-affinity relationship for describing rates of hydrocarbon metabolism.the kinetics of concentration-dependent toluene metabolism were examined by evaluating each term in the second-order rate equation. marine and freshwater pseudomonads were used. uptake for pseudomonas sp. strain t2 was characterized by a completely saturatable system with small transport constant (k(t) = 44 mug/liter) and large specific affinity. kinetics for pseudomonas putida ppf1 were similar. induction had little effect on k(t), but it caused the specific affinity to increase from about 0.03 ...198716347440
the effect of pseudomonas putida colonization on root surface peroxidase.increased activities of peroxidase and indole 3-acetic acid (iaa) oxidase were detected on root surfaces of bean (phaseolus vulgaris) seedlings colonized with a soil saprophytic bacterium, pseudomonas putida. iaa oxidase activity increased over 250-fold and peroxidase 8-fold. enhancement was greater for 6-day-old than for 4- or 8-day-old inoculated plants no iaa oxidase or peroxidase activities were associated with the bacterial cells. native polyacrylamide gel electrophoresis demonstrated that ...198716665732
comparison of a gelation and a chromogenic limulus (lal) assay for the detection of gram-negative bacteria, and the application of the latter assay to milk.when a chromogenic limulus amoebocyte lysate (lal) assay and a tube gelation lal assay were compared for the detection of gram-negative bacteria using a strain of pseudomonas putida, the detection level (approximately 10(3) cfu/ml) and cost of the assays were approximately the same for both assays but the reading was more precise for the chromogenic substrate assay. a modified chromogenic assay was devised for detection of ps. putida in milk.19873597923
amidohydrolysis of n-methylhydantoin coupled with atp hydrolysis.a new enzyme, n-methylhydantoin amidohydrolase, was highly purified from pseudomonas putida 77: it catalyzes the hydrolysis of n-methylhydantoin to n-carbamoylsarcosine with the concomitant stoichiometric cleavage of atp to adp and orthophosphate. the enzyme absolutely requires atp, mg2+ and k+ for the n-methylhydantoin hydrolysis. the rapid and complete degradation of n-methylhydantoin during the cultivation of p. putida 77, which rapidly degrades creatinine via only n-methylhydantoin and which ...19873827889
18o studies on the 5-oxoprolinase reaction. evidence for a phosphorylated tetrahedral intermediate.5-oxoprolinase catalyzes a reaction in which the cleavage of atp to adp and pi and the decyclization of 5-oxoproline to form glutamate are coupled. when the reaction catalyzed by 5-oxoprolinase of pseudomonas putida was carried out to 90% completion in h2(18)o, the residual 5-oxoproline was found to contain 18o in the amide carbonyl oxygen atom. such isotopic incorporation was not observed in similar studies with a subunit of the enzyme which catalyzes 5-oxoproline-dependent atpase and formation ...19873611103
hydrodynamic characterization of the size and shape of atropinesterase from pseudomonas putida.atropinesterase from pseudomonas putida has been investigated by means of different ultracentrifugation methods under native and denaturing conditions. the following quantities were determined: sedimentation coefficient, translational diffusion and friction coefficient, partial specific volume and molecular weight. from these data the size, shape and hydration of the enzyme molecule in solution were estimated. the results suggest that atropinesterase is a globular protein which consists of a sin ...19873828356
structure of the pseudomonas putida alkbac operon. identification of transcription and translation products.the structural genes of the pseudomonas oleovorans alk (alkane utilization) system, which are localized on the alkbac operon, were cloned as a 16.9-kilobase pair ecori fragment. we have measured the length and determined the position of the alkbac operon on this fragment by electron microscopy of r-loops. furthermore, the 7.3-kilobase pair long alkbac operon was analyzed for translation products in escherichia coli minicells. using a spectrum of overlapping subclones, six different proteins were ...19873032966
[automated micromethod for the determination of the utilization of carbon sources by clinically significant pseudomonas species].the assimilation of 43 different carbon substrates by 93 clinical strains of pseudomonas aeruginosa was studied by a new miniaturized rapid method. reading of assimilation results was done photometrically after 18-20 h incubation and the resulting data were captured and stored by a microcomputer. the differentiating capacity of the assimilation tests were verified by comparing the results of 41 strains of pseudomonas fluorescens, 48 strains of pseudomonas putida, 52 strains of pseudomonas maltop ...19873118596
a set of cassettes and improved vectors for genetic and biochemical characterization of pseudomonas genes.a set of broad-host-range vectors allowing direct selection of recombinant dna molecules to facilitate subcloning and expression analyses of pseudomonas genes was constructed using bg/ii lacz alpha cassette. controlled expression vectors pvdtac39 and pvdtac24 were shown to be useful for determination of enzymatic activities encoded by the cloned dna fragments and mr determination of the corresponding polypeptides. a set of pseudomonas putida xyle gene cassettes truncated at the 5' end was constr ...19873123327
construction of a transposon containing a gene for polygalacturonate trans-eliminase from klebsiella oxytoca.a dna fragment containing a klebsiella oxytoca gene for polygalacturonate trans-eliminase was cloned into the kanamycin resistance transposon tn5. this new transposon, designated tn5-pga+, had a transposition frequency of 1 x 10(-6). the broad host range plasmid pr751::tn5-pga+ was conjugally transferred to a variety of genetic backgrounds. the ability to degrade polygalacturonate was expressed in aeromonas hydrophila, alcaligenes eutrophus, azotomonas insolita, escherichia coli, pseudomonas put ...19873034186
interposon mutagenesis of soil and water bacteria: a family of dna fragments designed for in vitro insertional mutagenesis of gram-negative bacteria.we have constructed a series of derivatives of the omega interposon [prentki and krisch, gene 29 (1984) 303-313] that can be used for in vitro insertional mutagenesis. each of these dna fragments carries a different antibiotic or hg2+ resistance gene (apr, cmr, tcr, kmr or hgr) which is flanked, in inverted orientation, by transcription and translation termination signals and by synthetic polylinkers. the dna of these interposons can be easily purified and then inserted, by in vitro ligation, in ...19873038679
purification and properties of formylglutamate amidohydrolase from pseudomonas putida.formylglutamate amidohydrolase (fgase) catalyzes the terminal reaction in the five-step pathway for histidine utilization in pseudomonas putida. by this action, n-formyl-l-glutamate (fg) is hydrolyzed to produce l-glutamate plus formate. urocanate, the first product in the pathway, induced all five enzymes, but fg was able to induce fgase alone, although less efficiently than urocanate did. this induction by fg resulted in the formation of an fgase with electrophoretic mobility identical to that ...19873308850
[molecular genetic organization and origin of plasmid pbs52 with a broad range of bacterial hosts].the data are presented on the localization of genetic determinants of resistance to streptomycin, ampicillin and sulfanilamides on the physical map of conjugative r plasmid pbs52 of 38,000 bp which has a broad bacterial host range and belongs to a new incompatibility group. the plasmid has a natural "polylinker" site (less than 200 bp) containing (in the order of arrangement) the recognition sites for restriction enzymes: bamhi-ecori-psti-ecorv-bglii (pvuii). the comparative analysis shows that ...19873319772
survival of selected bacterial species in sterilized activated carbon filters and biological activated carbon filters.the survival of selected hygienically relevant bacterial species in activated carbon (ac) filters on a bench scale was investigated. the results revealed that after inoculation of the test strains the previously sterilized ac absorbed all bacteria (10(6) to 10(7)). after a period of 6 to 13 days without countable bacteria in the effluent, the numbers of escherichia coli, pseudomonas aeruginosa, and pseudomonas putida increased up to 10(4) to 10(5) cfu/ml of effluent and 10(6) to 10(7) cfu/g of a ...19873579281
construction of chlorobenzene-utilizing recombinants by progenitive manifestation of a rare event.separate continuous cultures of pseudomonas putida r5-3, grown on toluene, and pseudomonas alcaligenes c-o, grown on benzoate, were concentrated and continuously amalgamated on a ceramic bead column, which was subjected to a continuous stream of chlorobenzene vapors. a recombinant strain, p. putida cb1-9, was isolated in less than 1 month. p. alcaligenes c-0 grew on benzoate and 3-chlorobenzoate but not on toluene, p. putida r5-3 grew on benzoate and toluene but not on 3-chlorobenzoate, and neit ...19873426217
effect of temperature on the stability of plasmid ptg201 and productivity of xyle gene product in recombinant escherichia coli: development of a two-stage chemostat with free and immobilized cells.the effect of temperature on the stability of ptg201, a plasmid carrying the xyle gene (which encodes catechol 2,3-dioxygenase from pseudomonas putida), and the production of catechol 2,3-dioxygenase in free and immobilized escherichia coli during continuous culture have been studied at various temperatures. immobilization of cells increased the stability of ptg201 considerably, even under conditions when expression of the xyle product was enhanced. since xyle transcription was controlled by the ...19873312486
molecular analysis of regulatory and structural xyl genes of the tol plasmid pww53-4.pww53-4 is a cointegrate between rp4 and the catabolic plasmid pww53 from pseudomonas putida mt53, which contains 36 kbp of pww53 dna inserted close to the oriv gene of rp4; it encodes the ability to grow on toluene and the xylenes, characteristic of pww53, as well as resistance to tetracycline, kanamycin and carbenicillin, characteristic of rp4. a physical map of the 36 kbp insert of pww53 dna for 11 restriction enzymes is presented, showing that the relative positions of the two xyl operons ar ...19873309179
laboratory and clinical evaluation of isolation media for campylobacter jejuni.six selective isolation media were evaluated for their ability to support the growth of campylobacter jejuni. colony counts of 70 isolated strains of c. jejuni and recovery studies on these strains in simulated positive feces samples demonstrated that bolton and hutchinson' charcoal, cefoperazone, deoxycholate agar and karmali's charcoal-based selective medium produced the highest recovery rates with the greatest suppression of other fecal flora. c. jejuni colonies were more easily recognized on ...19873429621
genetic analysis of mannityl opine catabolism in octopine-type agrobacterium tumefaciens strain 15955.the genetic organization of functions responsible for mannityl opine catabolism of the ti plasmid of agrobacterium tumefaciens strain 1,5955 was investigated. a partial hindiii digest of pti1,5955 was cloned into a broad host range cosmid and the clones obtained were tested for ability to confer mannityl opine degradation upon agrobacterium. inserts containing genes for catabolism of mannopinic acid, mannopine, agropine, and agropinic acid were obtained, spanning a segment of 43 kb on the ti pla ...19873112522
[expression of the human interferon alpha f gene in the obligate methylotroph methylobacillus flagellatum kt and pseudomonas putida].the expression of human leucocyte interferon alpha f gene in plasmid plm-ifn alpha f-273 is controlled by a hybrid tac (trp-lac) promoter. a structural gene for interferon alpha f is a component of the hybrid operon lacz'-ifn alpha f-tcr, that contains an e. coli trp-operon intercystronic region. plasmid plm ifn alpha f-273--directed interferon synthesis allows to obtain about 10(7) iu/l. this plasmid was cloned in broad-host-range vector plasmid payc31. the hybrid bi-repliconed plasmid containi ...19873119998
maintenance and stability of introduced genotypes in groundwater aquifer material.three indigenous groundwater bacterial strains and pseudomonas putida harboring plasmids tol (pwwo) and rk2 were introduced into experimentally contaminated groundwater aquifer microcosms. maintenance of the introduced genotypes was measured over time by colony hybridization with gene probes of various specificity. on the basis of the results of colony hybridization quantitation of the introduced organisms and genes, all introduced genotypes were stably maintained at approximately 10(5) positive ...19873300546
organization and nucleotide sequence determination of a gene cluster involved in 3-chlorocatechol degradation.three critical enzymes catechol oxygenase ii (chlorocatechol dioxygenase), muconate cycloisomerase ii, and dienelactone hydrolase, are involved in the degradation of chlorocatechols, which are obligatory intermediates in the catabolism of chlorinated aromatic compounds. the organization and complete nucleotide sequence of the genes for these enzymes have been determined on a 4.2-kilobase-pair (kbp) bgl ii fragment cloned from the plasmid pac27, based on the agreement of open reading frame length ...19873299368
activation of the xyldlegf promoter of the tol toluene-xylene degradation pathway by overproduction of the xyls regulatory gene product.the xyls regulatory gene of the pseudomonas putida tol plasmid (pwwo) has been cloned under the transcriptional control of the escherichia coli tac promoter in a broad-host-range controlled-expression vector. induction with isopropylthiogalactoside allowed overproduction and characterization of the xyls product by specific interaction with the tol meta-cleavage pathway operator-promoter region (op2) in vivo in e. coli. examination of plasmid-specified polypeptides in e. coli maxicells led to ide ...19873301806
molecular cloning of genes encoding branched-chain keto acid dehydrogenase of pseudomonas putida.we cloned the structural genes for the individual subunits of the branched-chain keto acid dehydrogenase multienzyme complex on a 7.8-kilobase ecori-ssti restriction fragment of pseudomonas putida chromosomal dna by cloning into the broad-host-range vector pkt230. a direct selection system for growth on valine-isoleucine agar was achieved by complementation of p. putida branched-chain keto acid dehydrogenase mutants. the recombinant plasmid, pss1-1, increased expression of branched-chain keto ac ...19873549697
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