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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
pseudomonas putida. newly recognized pathogen in patients with cancer.pseudomonas putida was recovered from blood culture specimens between 1980 and 1985 in 15 patients with cancer. no isolates were found in specimens obtained before 1980. eight patients were considered to have septicemia (more than one positive blood culture result plus clinical signs of infection). septicemia was monomicrobial in three of those eight patients and polymicrobial in five. of these eight patients, one had pneumonia and three had phlebitis, cellulitis, or both at the site of the veno ...19873605136
cloning and complete nucleotide sequence determination of the catb gene encoding cis,cis-muconate lactonizing enzyme.the enzyme, cis,cis-muconate lactonizing enzyme i (mlei; ec 5.5.1.1), has been proposed to play a key role in the beta-ketoadipate pathway of benzoate degradation. a 10.2-kb ecori fragment isolated from a pseudomonas putida genomic library complemented a mutant deficient in this enzyme. the mlei coding gene, catb, was localized to a 1.6-kb fragment which was sequenced by the dideoxy chain termination method. mlei was purified 25-fold from crude extracts of benzoate-grown p. putida prs2015 harbor ...19873609743
overproduction of the xyls gene product and activation of the xyldlegf operon on the tol plasmid.the effect of high-level expression of the regulatory gene xyls of the pseudomonas putida tol plasmid on the activation of the xyldlegf operon was investigated in escherichia coli. the xyls gene was placed downstream from the tac promoter, and the resultant fusion was cloned in cis to the xyldlegf operon. the expression of the operon was monitored by the level of catechol 2,3-dioxygenase, whose structural gene xyle was placed directly after the operator-promoter region of xyldlegf. xyls transcri ...19873611023
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
high-resolution crystal structure of cytochrome p450cam.the crystal structure of pseudomonas putida cytochrome p450cam with its substrate, camphor, bound has been refined to r = 0.19 at a normal resolution of 1.63 a. while the 1.63 a model confirms our initial analysis based on the 2.6 a model, the higher resolution structure has revealed important new details. these include a more precise assignment of sequence to secondary structure, the identification of three cis-proline residues, and a more detailed picture of substrate-protein interactions. in ...19873656428
nucleotide sequencing and characterization of the genes encoding benzene oxidation enzymes of pseudomonas putida.the nucleotide sequence of the genes from pseudomonas putida encoding oxidation of benzene to catechol was determined. five open reading frames were found in the sequence. four corresponding protein molecules were detected by a dna-directed in vitro translation system. escherichia coli cells containing the fragment with the four open reading frames transformed benzene to cis-benzene glycol, which is an intermediate of the oxidation of benzene to catechol. the relation between the product of each ...19873667527
the effect of lipophilic weak acids on the segregational stability of tol plasmids in pseudomonas putida.the effect of various lipophilic weak acids on the stability of certain tol plasmids was investigated. benzoate induced deletion of tol plasmid dna in pseudomonas putida mt15, followed by loss of the plasmid; this effect was ph- and concentration-dependent, suggesting that undissociated benzoic acid was a more effective curing agent than the benzoate anion. plasmid loss always approached a frequency of 100% after a lag and apparently depended on the prior occurrence of deletions, although delete ...19873668501
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
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
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
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 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
qualitative evidence for expression of klebsiella pneumoniae nif in pseudomonas putida.pseudomonas putida mt20-3 carrying the klebsiella pneumoniae nif plasmids prd1 or pmf250 showed highly o2-sensitive aerobic acetylene reduction on low-n pyruvate or glucose agar. this finding confirms unequivocally that k. pneumoniae nif can be expressed in an obligate aerobe.19873329216
[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
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
the amino acid sequence of the aliphatic amidase from pseudomonas aeruginosa.amino acid sequence studies show that the aliphatic amidase (ec 3.5.1.4) from pseudomonas aeruginosa pac142 consists of a single polypeptide chain of 346 residues, giving an mr of 38,400. the evidence from the amino acid studies is in complete agreement with that deduced from the dna sequence of the amie gene. studies of the protein from pseudomonas putida a87 show that it differs from the ps. aeruginosa protein by about 30 amino acid substitutions. it now becomes possible to relate changes in t ...19873108029
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
[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
gene organization of the first catabolic operon of tol plasmid pww53: production of indigo by the xyla gene product.the entire operon coding for the enzymes responsible for conversion of toluenes to benzoates has been cloned from tol plasmid pww53 and the position of the genes accurately located. the coding region was 7.4 kilobase pairs (kbp) long, and the gene order was operator-promoter region (op1)-a small open reading frame-xylc (1.6 kbp)-xyla (2.9 kbp)-xylb (1.8 kbp). within the coding region there was considerable homology with the isofunctional region of the archetypal tol plasmid pww0. a central regio ...19873027047
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
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
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
effect of temperature on sulfite-mediated dark reversion of urocanase in pseudomonas putida. 19872888143
controlled and functional expression of the pseudomonas oleovorans alkane utilizing system in pseudomonas putida and escherichia coli.the oct plasmid encodes enzymes for alkane hydroxylation and alkanol dehydrogenation. structural components are encoded on the 7.5-kilobase pair alkbac operon, whereas positive regulatory components are encoded by alkr. we have constructed plasmids containing fusions of cloned alkbac and alkr dna and used these fusion plasmids to study the functional expression of the alkbac operon and the regulatory locus alkr in pseudomonas putida and in escherichia coli. growth on alkanes requires a functiona ...19872826430
isolation and characterization of pseudomonas putida r-prime plasmids.a number of enhanced chromosome mobilizing (ecm) plasmids derived from the wide host range plasmid r68 have been used to construct r-prime plasmids carrying a maximum of two map minutes of the pseudomonas putida ppn chromosome, using pseudomonas aeruginosa pao as the recipient. for one ecm plasmid, pmo61, the ability to form r-primes did not correlate with the ability to mobilize chromosomes in intrastrain crosses, suggesting that different mechanisms are involved. physical analysis of one r-pri ...19872821167
[bacterial destruction of anionic sulfoethoxylate surfactants].a pseudomonas putida strain g was isolated and its activity in the destruction of sulfoethoxylates (surfactants) was studied as a function of the cultivation conditions. ultrastructural changes were found in the cells utilizing sulfoethoxylates. sulfoethoxylates were found to be decomposed by p. putida g via two pathways: (a) desulfation of the molecule and (b) cleavage of the alkyl ester bond. the intermediate products were not toxic and did not pollute the habitat. the strain can be used for m ...19872826975
phosphate-selective porins from the outer membranes of fluorescent pseudomonas sp.phosphate starvation induced oligomeric proteins from the outer membranes of pseudomonas fluorescens, pseudomonas putida, pseudomonas aureofaciens, and pseudomonas chlororaphis were purified to homogeneity. the incorporation of the purified proteins into planar lipid bilayer membranes resulted in stepwise increases in membrane conductance. single channel conductance experiments demonstrated that these proteins were all capable of forming small channels, similar to the pseudomonas aeruginosa phos ...19872436733
expression of the regulatory gene xyls on the tol plasmid is positively controlled by the xylr gene product.the regulatory gene xyls on the tol plasmid of pseudomonas putida activates the transcription of the xyldlegf operon for the m-toluate-degrading pathway in the presence of m-toluate. the gene also activates the transcription of the same operon in the presence of m-xylene or m-methylbenzyl alcohol, but for this activation another regulatory gene, xylr, is required. in this study we examined the xyls expression by determining the mrna by reverse transcriptase mapping and by monitoring the enzyme a ...19872440045
analysis of transcription from the trfa promoter of broad host range plasmid rk2 in escherichia coli, pseudomonas putida, and pseudomonas aeruginosa.reverse transcriptase mapping has been used to analyze transcription from the trfa promoter of broad host range plasmid rk2. the results show that trfa operon mrna has the same 5' end in pseudomonas aeruginosa, pseudomonas putida, and escherichia coli. the strengths of wild-type and mutant trfa promoters, which differ by defined base substitutions, have been compared and the positions of their transcriptional start sites determined. while these base substitutions do not alter the transcriptional ...19872442786
purification and characterization of adhesive exopolysaccharides from pseudomonas putida and pseudomonas fluorescens.in this study, the adhesive exopolysaccharides of strains of pseudomonas putida and p. fluorescens, both isolated from freshwater epilithic communities, were examined with regard to their chemical composition, biosynthesis, and their role in adhesion. electron microscopy showed that both strains were enrobed in fibrous glycocalyces and that these structures were involved in attachment of the cells to a solid surface and as structural matrices in the microcolony mode of growth. in batch culture e ...19872451553
molecular cloning of the pseudomonas putida glyoxalase i gene in escherichia coli.the glyoxalase i gene of pseudomonas putida was cloned onto a vector plasmid pbr 322 as a 7.5 kilobase sau 3ai fragment of chromosomal dna and the hybrid plasmid was designated pgi 318. the gene responsible for the glyoxalase i activity in pgi 318 was recloned in pbr 322 as a 2.2 kilobase hin diii fragment and was designated pgi 423. the p. putida glyoxalase i gene on pgi 318 and pgi 423 was highly expressed in e. coli cells and the glyoxalase i activity level was increased more than 150 fold in ...19872820418
in vivo formation of gene fusions in pseudomonas putida and construction of versatile broad-host-range vectors for direct subcloning of mu d1 and mu d2 fusions.the mu d1 and mu d2 prophages were integrated into the conjugative broad-host-range plasmid r751. the two plasmids were then transferred into pseudomonas putida, and derivatives carrying intact mu prophages were recovered. after induction of mu at 42 degrees c, both operon and gene fusions were observed on 5-bromo-4-chloro-3-indolyl-beta-d-galactopyranoside (x-gal) plates. broad-host-range vectors were constructed which allow direct cloning of both operon or gene fusions and their analysis in es ...19872821901
electron paramagnetic resonance study of ferrous cytochrome p-450scc-nitric oxide complexes: effects of cholesterol and its analogues.electron paramagnetic resonance (epr) spectra of nitric oxide (no) complexes of ferrous cytochrome p-450scc were measured at 77 k for the first time without using the rapid-mixing and freeze-quenching technique. without substrate the epr spectra were very similar to those of cytochrome p-450cam (from pseudomonas putida) and cytochrome p-450lm (from rat liver microsomes) with rhombic symmetry; gx = 2.071, gz = 2.001, gy = 1.962, and az = 2.2 mt for 14no complexes. upon addition of substrates [suc ...19872822097
comparison of the spin-lattice relaxation properties of the two classes of [2fe-2s] clusters in proteins.two classes of [2fe-2s] proteins have been defined according to the mean value gav of their g tensor components (bertrand, p., guigliarelli, b., gayda, j.p., beardwood, p. and gibson, j.f. (1985) biochim. biophys. acta 831, 261-266). to characterize their magnetic properties better, we have compared the spin-lattice relaxation behavior of typical proteins which belong to these two classes, namely spirulina maxima and adrenal ferredoxin for the gav approximately 1.96 class, thermus thermophilus r ...19872822125
effect of a broad-host range plasmid on growth dynamics of escherichia coli and pseudomonas putida.host-plasmid interactions were studied for the broad-host range plasmid, ptjs26, a derivative of rk2. to isolate host and plasmid contributions to the growth dynamics and plasmid stability, separate experiments were performed with host and recombinant cells for two different gram-negative hosts, pseudomonas putida and escherichia coli, at two different temperatures, 30 and 37 degrees c. at the lower temperature (30 degrees c) the growth kinetics were not affected by the plasmid, but plasmid inst ...198718576486
production of toluene cis-glycol by pseudomonas putida in glucose feb-batch culture.toluene was oxidized by a mutant strain of pseudomonas putida (strain ng1) to toluene cis-glycol (tcg). product was accumulated in fed-batch cultures to concentrations (18-24 g/l) higher than hitherto achieved. in vitro activities of toluene dioxygenase from p. putida ng1 were fivefold lower than that from the toluene-grown wild-type organism, whereas comparable activities of both catechol 2,3- and catechol 1,2-oxygenase were obtained; irreversible inhibition of toluene dioxygenase activity by t ...198718576532
air-stripping effects on cell growth with volatile substrates.the removal of substate molecules from aerobic microbial cultures is due to both consumption by microorganisms and stripping by the air stream. the air stripping component can be described by a constant parameter for low concentrations of volatile substrates. this air stripping parameter was found to have a value of 0.0033 h(-1) for phenol molecules in a typical fermentation situation. the determination and inclusion of this constant is important for modeling microbial growth. for pseudomonas pu ...198718581430
oxidation of an inhibitory substrate by washed cells (oxidation of phenol by pseudomonas putida).the specific uptake rate of phenol by washed cells of pseudomonas putida grown on phenol in steady-state continuous culture at various dilution rates was studied. the monod-haldane-type equation was applied to fit the data and the best kinetic parameters were determined by nonlinear least-squares techniques. the values of the kinetic parameters were found to increase monotonically with the phenol concentration in the original chemostat. the relations between the values of kinetic parameters and ...198718581530
biodehalogenation of bromotrichloromethane and 1,2-dibromo-3-chloropropane by pseudomonas putida ppg-786.biodehalogenation of 10(-5) m concentrations of bromotrichloromethane (btm) and 1,2-dibromo-3-chloropropane (dbcp) was studied in static cultures of pseudomonas putida ppg-786. the experimental cultures were prepared by growing p. putida on camphor, which is known to induce the synthesis of high concentrations of cytochrome p-450 in this bacterium. measurements of bromide ion release were found to be approximately consistent with the amounts of halocarbon degraded. gas chromatography/elctron cap ...198718576369
continuous production of l-phenylalanine by transamination.l-phenylalanine was produced continuously from l-as-partate and phenylpyruvate by transaminase from a newly screened pseudomonas putida strain. the process was carried out with an isolated enzyme in homogeneous phase in an enzyme membrane reactor and with immobilized whole cells in a stirred tank reactor, respectively. due to the difference in transport resistance, the productivity of the free enzyme in homogeneous phase (72 mmol/l h) was about 3 times higher than the productivity achieved using ...198718576476
dynamics of continuous stirred-tank biochemical reactor utilizing inhibitory substrate.the model of a continuous-stirred tank biochemical reactor was developed in which the instant uptake rate of substrate was used. the solutions of the model found for the oxidation of phenol by pseudomonas putida fitted the experimental data better than the results obtained from the models cited in the literature. the model enables control of the culture parameters so that the unwanted washout of the biomass from the bioreactor can be avoided. a review of the models cited in the literature is als ...198818584592
uptake rate of phenol by pseudomonas putida grown in unsteady state.the uptake rate of phenol by washed cells of pseudomonas putidagrown on phenol in fermenter in an un steady state, caused by the step increase of dilution rate and/or phenol concentration in the feed, was studied. the monod-haldane type equation was applied to fit the data and the best kinetic parameters were calculated by nonlinear least-square techniques. it was found that the minimum period of unsteady state required for induction of the phenol metabolic pathway was approximately 30 min. the ...198818587828
radiometric determination of uranium accumulated in bacterial cells.a method for the radiometric determination of uranium accumulated in bacterial cells was investigated. pseudomonas putida atcc 33015 was grown in a medium containing uranium in the form of uo(2)(no(3))(2). the cells were harvested by centrifugation, washed, resuspended in a buffer solution, ultrasonically disrupted and then suspended in unisolve-1. the radiation emitted by natural uranium isotopes (mainly (238)u and (234)u) was measured by liquid scintillation counting with natural uranium as an ...198818964500
nucleotide homology and organization of chlorocatechol oxidation genes of plasmids pjp4 and pac27.the 2,4-dichlorophenoxyacetate (2,4-d) catabolic plasmid pjp4 of alcaligenes eutrophus jmp134 contains two sets of nonidentical chlorocatechol oxidation gene sequences physically separated by a 7 kb dna region. we determined the nucleotide sequence of the 1.6 kb hindiii fragment containing the known genes tfdc and tfdd (don et al. 1985) which encode pyrocatechase and cycloisomerase, respectively. the 1.3 kb bglii-hindiii segment of recombinant plasmid pdc25 containing at least three chlorocatech ...19882830460
cloning, dna sequence analysis, and expression in escherichia coli of the gene for mandelate racemase from pseudomonas putida.the gene for mandelate racemase (ec 5.1.2.2) from pseudomonas putida (atcc 12633) was cloned in pseudomonas aeruginosa (atcc 15692). the selection for the cloned gene was based upon the inability of p. aeruginosa to grow on (r)-mandelate as sole carbon source by virtue of the absence of mandelate racemase in its mandelate pathway. fragments of p. putida dna obtained by digestion of chromosomal dna with sau3a were ligated into the bamhi site of the gram-negative vector pkt230 and transformed into ...19882831968
variation in the ability of pseudomonas sp. strain b13 cultures to utilize meta-chlorobenzoate is associated with tandem amplification and deamplification of dna.single-colony isolates of pseudomonas sp. strain b13 were examined for their ability to utilize benzoate (ben) and meta-chlorobenzoate (3cb) as the sole carbon source. scoring of b13 cultures by the replica-plating technique revealed that under nonselective conditions, b13 spontaneously formed four different types of colonies: 3cb+ ben+, 3cb+ ben-, 3cb- ben-, 3cb- ben+. successive testing of each of the four colony types showed that each produced the same four different types of single-colony is ...19882832387
variation in the ability of pseudomonas sp. strain b13 cultures to utilize meta-chlorobenzoate is associated with tandem amplification and deamplification of dna.single-colony isolates of pseudomonas sp. strain b13 were examined for their ability to utilize benzoate (ben) and meta-chlorobenzoate (3cb) as the sole carbon source. scoring of b13 cultures by the replica-plating technique revealed that under nonselective conditions, b13 spontaneously formed four different types of colonies: 3cb+ ben+, 3cb+ ben-, 3cb- ben-, 3cb- ben+. successive testing of each of the four colony types showed that each produced the same four different types of single-colony is ...19882832387
transcription of the fimbrial subunit gene and an associated transfer rna gene of pseudomonas aeruginosa.gene fima encoding the structural subunit of the fimbriae of pseudomonas aeruginosa pak is located in the centre of a 1.2-kb hindiii genomic dna fragment [see also sastry et al., j. bacteriol. 164 (1985) 571-577], which in turn is located within a 6.2-kb ecori fragment. immediately downstream from fima is a putative threonine trna gene [dalrymple and mattick, biochem. int. 13 (1986) 547-553]. northern blotting experiments showed that fima is transcribed to an mrna of approx. 650 nucleotides, whi ...19882452767
transcriptional regulation, nucleotide sequence, and localization of the promoter of the catbc operon in pseudomonas putida.the catb and catc genes encode cis,cis-muconate lactonizing enzyme i (ec 5.5.1.1) and muconolactone isomerase (ec 5.3.3.4), respectively. these enzymes are required for the dissimilation of benzoate to beta-ketoadipate by pseudomonas putida and are under coordinate transcriptional regulation. by deletion analysis and the use of pkt240 as a promoter probe vector, we located a single promoter region for the catbc operon upstream of catb. rna-dna hybridization studies, together with reverse transcr ...19882449420
genetic organization and transcriptional analysis of a major gene cluster involved in siderophore biosynthesis in pseudomonas putida wcs358.in iron-limited environments, the plant-growth-stimulating pseudomonas putida wcs358 produces a yellow-green fluorescent siderophore called pseudobactin 358. the transcriptional organization and the iron-regulated expression of a major gene cluster involved in the biosynthesis and transport of pseudobactin 358 were analyzed in detail. the cluster comprises a region with a minimum length of 33.5 kilobases and contains at least five transcriptional units, of which some are relatively large. the di ...19882450869
organization and multiple regulation of histidine utilization genes in pseudomonas putida.the arrangement of the histidine utilization (hut) genes in pseudomonas putida was established by examining the structure of a dna segment that had been cloned into escherichia coli via a cosmid vector. southern blot analysis revealed that the restriction patterns of the hut genes cloned into e. coli and present in the p. putida genome were identical, indicating that no detectable dna rearrangement took place during the cloning. expression of the hut genes from a series of overlapping clones ind ...19882842309
alkane utilization in pseudomonas oleovorans. structure and function of the regulatory locus alkr.the oct plasmid-localized alkbac operon encodes enzymes for alkane hydroxylation and alkanol dehydrogenation. the positively controlled expression of the operon is very efficient in both pseudomonas putida and escherichia coli. two regulatory functions have been ascribed to the regulatory locus alkr: inducer recognition and transcriptional activation of the operon. we have cloned and localized the alkr locus on a 4.9-kilobase pair sali fragment. the alkr region was analyzed for translation produ ...19882843518
enumeration of tn5 mutant bacteria in soil by using a most- probable-number-dna hybridization procedure and antibiotic resistance.investigations were made into the utility of dna hybridization in conjunction with a microdilution most-probable-number procedure for the enumeration of rhizobium spp. and pseudomonas putida in soil. isolates of rhizobium spp. and p. putida carrying the transposon tn5 were added to sterile and nonsterile burbank sandy loam soil and enumerated over time. soil populations of rhizobia were enumerated by colony hybridization, most-probable-number-dna hybridization procedure, plate counts, plant infe ...19882833161
toluene degradation by pseudomonas putida f1: genetic organization of the tod operon.pseudomonas putida ppf1 degrades toluene through cis-toluene dihydrodiol to 3-methylcatechol. the latter compound is metabolized through the well-established meta pathway for catechol degradation. the first four steps in the pathway involve the sequential action of toluene dioxygenase (todabc1c2), cis-toluene dihydrodiol dehydrogenase (todd), 3-methylcatechol 2,3-dioxygenase (tode), and 2-hydroxy-6-oxo-2,4-heptadienoate hydrolase (todf). the genes for these enzymes form part of the tod operon wh ...19882843094
cosmid cloning of five zymomonas trp genes by complementation of escherichia coli and pseudomonas putida trp mutants.a library of zymomonas mobilis genomic dna was constructed in the broad-host-range cosmid plafr1. the library was mobilized into a variety of escherichia coli and pseudomonas putida trp mutants by using the helper plasmid prk2013. five z. mobilis trp genes were identified by the ability to complement the trp mutants. the trpf, trpb, and trpa genes were on one cosmid, while the trpd and trpc genes were on two separate cosmids. the organization of the z. mobilis trp genes seems to be similar to th ...19882838460
benzoate-dependent induction from the op2 operator-promoter region of the tol plasmid pwwo in the absence of known plasmid regulatory genes.expression of the lower catabolic pathway of the tol plasmid pwwo requires an aromatic acid inducer and the product of the xyls regulatory gene. pseudomonas putida cells transformed with a plasmid containing the operator-promoter region of the lower pathway (op2 [or pm]), upstream from the catechol 2,3-dioxygenase structural gene, showed enzyme induction in the absence of known tol plasmid regulatory genes. induction was not seen in transformed escherichia coli cells or in a p. putida mutant lac ...19882841300
klebsiella pneumoniae origin of replication (oric) is not active in caulobacter crescentus, pseudomonas putida, and rhodobacter sphaeroides.a dna fragment carrying genes encoding the conjugal transfer system of the broad host range plasmid rk2 was inserted into a plasmid carrying the chromosomal origin of replication (oric) from klebsiella pneumoniae. the resulting plasmid, peon1, was readily transferred between gram-negative bacteria and carried two potential origins of replication: oric and the replication origin from pbr322 (oripbr). although peon1 could be transferred to caulobacter crescentus, pseudomonas putida, and rhodobacte ...19882841304
isolation and characterization of a new plasmid from a flavobacterium sp. which carries the genes for degradation of 2,4-dichlorophenoxyacetate.a flavobacterium sp. (strain 50001), capable of degrading 2,4-dichlorophenoxyacetate (2,4-d), 2-methyl-4-chlorophenoxyacetate, and 2-chlorobenzoate and imparting resistance to mercury, harbored a degradative plasmid, prc10. cured strains of the flavobacterium sp. lost the plasmid as well as the ability to degrade these chlorinated compounds. comparison of this plasmid with the well-characterized 2,4-d-degradative plasmid pjp4 from alcaligenes eutrophus showed regions of homology between the two ...19882842290
[interaction of heterologous bacteriophages: growth suppression of temperate pp56 phage of pseudomonas putida by the transposable phage d3112 of pseudomonas aeruginosa].we have found an inhibiting effect of hybrid rp4::d3112 plasmid (where d3112 is represented as genome of a transposable phage specific for pseudomonas aeruginosa) on the development of temperate p. putida phage pp56. the study of the effect has revealed a previously unknown locus (in the region 12-14.2 kb of the d3112 genome) which functions in the prophage state. the locus affects pp56 decreasing phage yield. mutants of pp56 insensitive to inhibition were found.19882843421
construction of an expression vector for the fission yeast schizosaccharomyces pombe.we have isolated and characterized a s. pombe promoter using a functional heterologous gene product assay. random s. pombe genomic fragments were cloned upstream from the promoterless 'lacz gene and tested in vivo for their efficiency to promote expression of the beta-galactosidase protein in the fission yeast. an efficient s. pombe promoter called 54/1 was isolated and shown to drive up to 5% of total protein synthesis as beta-galactosidase. the structure and nucleotide sequence of this promote ...19882843820
siderophore-mediated uptake of fe3+ by the plant growth-stimulating pseudomonas putida strain wcs358 and by other rhizosphere microorganisms.under iron-limited conditions, pseudomonas putida wcs358 produces a siderophore, pseudobactin 358, which is essential for the plant growth-stimulating ability of this strain. cells of strain wcs358, provided that they have been grown under fe3+ limitation, take up 55fe3+ from the 55fe3+-labeled pseudobactin 358 complex with km and vmax values of 0.23 microm and 0.14 nmol/mg of cell dry weight per min, respectively. uptake experiments with cells treated with various metabolic inhibitors showed th ...19882971647
[cloning and localization of the replication and mobilization regions in the d-plasmid of pseudomonas putida pbs286 (incp-9) with a broad host range].rep-mob loci of naphthalene degradative plasmid pbs286 (incp-9) have been cloned on the escherichia coli vectors puc19 and pubr322. these loci confer to recombinant plasmids pbs952 and pbs953 the ability for effective mobilization by rp4 (incp-1) and f plasmid, as well as constant maintenance in various gram-negative bacteria. localization of cloned sequences in the restriction fragments of conservative part of the pbs286 genome was established. the data obtained correlate with the analysis of p ...19882844624
purification and characterization of cutinase from a fluorescent pseudomonas putida bacterial strain isolated from phyllosphere.cutinase, an extracellular enzyme, was induced by cutin in a fluorescent pseudomonas putida strain that was found to be cohabiting with an apparently nitrogen-fixing corynebacterium. this enzyme was purified from the culture fluid by acetone precipitation followed by chromatography on deae-cellulose, qae-sephadex, sepharose 6b, and sephadex g-100. the purified enzyme showed a single band when subjected to polyacrylamide electrophoresis and the enzymatic activity coincided with the protein band. ...19883130804
[genes encoding bacterial rna-polymerase. i. cloning of rpobc-operon of pseudomonas putida and its physical map].the p. putida rpobc operon, coding for beta and beta' subunits of rna polymerase, was cloned and its physical map constructed.19883048271
pseudomonas stutzeri ferredoxin: close similarity to azotobacter vinelandii and pseudomonas ovalis ferredoxins.the complete primary structure of pseudomonas stutzeri strain zobell ferredoxin was determined by a combination of protease digestion, edman degradation, and carboxypeptidase digestion and was: tfvvtdncikckytdcvevcpvdcfyegpnflvih pdecidcalcepecpaqaifsedevpedqqefielnadlaevwpnite kkdaladaeewdgvkdklqyler. the calculated molecular weight was 12,110 excluding iron and sulfur atoms. the amino acid sequence was highly homologous to those of azotobacter vinelandii and pseudomonas ovalis ferredoxins. it ...19883053681
purification and properties of carnitine dehydrogenase from pseudomonas putida.carnitine dehydrogenase (carnitine:nad+ oxidoreductase, ec 1.1.1.108) from pseudomonas putida ifp 206 catalyzes the oxidation of l-carnitine to 3-dehydrocarnitine. the enzyme was purified 72-fold to homogeneity as judged by polyacrylamide gel electrophoresis. the molecular mass of this enzyme is 62 kda and consists of two identical subunits. the isoelectric point was found to be 4.7. the carnitine dehydrogenase is specific for l-carnitine and nad+. the optimum ph for enzymatic activity in the ox ...19883058208
molecular cloning of 3-phenylcatechol dioxygenase involved in the catabolic pathway of chlorinated biphenyl from pseudomonas putida and its expression in escherichia coli.genes encoding 3-phenylcatechol dioxygenases were cloned from the chlorobiphenyl-degrading pseudomonas putida strain ou83, using broad-host-range cosmid vector pcp13. restriction enzyme analysis of dna from 2,3-dioxygenase-positive chimeric cosmids showed dna inserts ranging in size from 6.0 to 30 kilobases. the origin of the dna insert in hybrid clones was established by using 32p-labeled hybrid clones (poh101 and poh810). a 2.3-kilobase hindiii fragment was common to two clones. the 2,3-dioxyg ...19883063207
[nucleotide sequence of of rpob gene encoding the rna-polymerase beta-subunit in pseudomonas putida]. 19883069413
the complete amino acid sequence and identification of the active-site arginine peptide of escherichia coli 2-keto-4-hydroxyglutarate aldolase.the complete amino acid sequence of 2-keto-4-hydroxyglutarate aldolase from escherichia coli has been established in the following manner. after being reduced with dithiothreitol, the purified aldolase was alkylated with iodoacetamide and subsequently digested with trypsin. the resulting 19 peptide peaks observed by high performance liquid chromatography, which compared with 21 expected tryptic cleavage products, were all isolated, purified, and individually sequenced. overlap peptides were obta ...19883136164
[nucleotide sequence of the rpoc-gene coding for rna polymerase beta'-subunit of pseudomonas putida]. 19883069416
cloning of genes for branched-chain keto acid dehydrogenase in pseudomonas putida. 19883071713
cytochrome p450: molecular architecture, mechanism, and prospects for rational inhibitor design.cytochromes p450 catalyze the insertion of one o2-derived oxygen atom into an aliphatic or aromatic molecule. p450s are best known for the metabolism of xenobiotic molecules, where hydroxylation renders insoluble hydrocarbons more soluble for easier elimination. in addition to this important catabolic function, p450s catalyze key steps in steroid and plant growth regulator metabolism. a variety of therapeutic, fungicidal, and agochemical agents that perturb these metabolic pathways very likely o ...19883073382
[bacteriophages of pseudomonas putida containing single-stranded canonical dna breaks].it was shown that bacteriophage tf as well as bacteriophages phi p4/40, phi p25/42, phi p23/40 and phi p6/40, which are specific to different p. putida strains, contain the single strand breaks in their dna. the breaks are localized in one strand of dna molecules and are repairable with t4 dna ligase. bacteriophage tf has no detectable dna homology with phi p4/40, phi p25/42, phi p23/40 and phi p6/40 bacteriophages. all the phages studied have no relation with other known pseudomonas phages. bac ...19883137462
comparison of the amino acid sequences of the transacylase components of branched chain oxoacid dehydrogenase of pseudomonas putida, and the pyruvate and 2-oxoglutarate dehydrogenases of escherichia coli.the nucleotide sequence of bkdb, the structural gene for e2b, the transacylase component of branched-chain-oxoacid dehydrogenase of pseudomonas putida has been determined and translated into its amino acid sequence. the start of bkdb was identified from the n-terminal sequence of e2b isolated from branched-chain-oxoacid dehydrogenase of the closely related species, p. aeruginosa. the reading frame was composed of 65.5% g + c with 82.3% of the codons ending in g or c. there was no intergenic spac ...19883046941
stability fluctuations of plasmid-bearing cells: immobilization effects.the maintenance of the plasmid vectors ptg201 and ptg206 (which both carry the pseudomonas putida xyle gene) and pb lambda h3 in escherichia coli hosts was studied in free and immobilized continuous cultures. ptg201, containing the strong lambda pr promoter, was more quickly lost than plasmid ptg206, containing the tetracycline resistance gene promoter. the instability of ptg201 seems to be related to high expression of the cloned xyle genet. fluctuations in the proportion of ptg201-containing c ...19883075659
construction of a shuttle vector for inducible gene expression in escherichia coli and bacillus subtilis.the construction of a shuttle vector for inducible gene expression allowing fast and easy cloning in escherichia coli and subsequent transformation of bacillus subtilis is presented. the expression is based on the regulation of the tac promoter by the lac repressor which was assayed with the xyle gene from pseudomonas putida as a marker gene. the laciq gene, transcribed by the strong spo promoter, allowed full repression of the weak tac promoter.19883141570
[isolation and comparative characteristics of 2 unrelated temperate phages of pseudomonas putida ppg1].two temperate bacteriophages, pp56 and pp71, specific for bacteria of pseudomonas putida strain ppg1 have been isolated for the first time. characterization of the phages was performed. both of them accomplish stable lysogenization of p. putida ppg1 cells. the phages are inducible. several groups of clear plaque (c) mutants of pp56 and pp71 with altered processes of establishment and maintenance of lysogenic state have been isolated, according to complementation test. the phages differ in follow ...19883129337
survival of rifampin-resistant mutants of pseudomonas fluorescens and pseudomonas putida in soil systems.the fate of spontaneous chromosomal rifampin-resistant (rifr) mutants of pseudomonas putida and pseudomonas fluorescens in sterile and live organic soil from which they were isolated was studied. in sterile native-soil assays, a rifr mutant of p. putida showed no decrease in competitive fitness when compared with the wild-type parent. however, mutants of p. fluorescens were of two general categories. group 1 showed no difference from the wild type in terms of growth rate, competitive fitness, an ...19883144244
[cloning and expression of pseudomonas putida gene controlling the catechol-2,3-oxygenase activity in escherichia coli cells].the genes nahc and nahd from pseudomonas putida naphthalene degradation plasmid pbs286 were cloned on the vector puc19 in escherichia coli cells. the catechol-2,3-oxygenase activity observed in e. coli cells containing recombinant plasmid pbs955 demands the participation of 32 kd polypeptide which is apparently the product of the nahc gene. second polypeptide of molecular weight 34.5 kd is synthesized in pbs955 containing e. coli minicells and perhaps it is a nahd gene product. the data obtained ...19883058550
cloning and expression of the cata and catbc gene clusters from pseudomonas aeruginosa pao.a 9.9-kilobase (kb) bamhi restriction endonuclease fragment encoding the cata and catbc gene clusters was selected from a gene bank of the pseudomonas aeruginosa pao1c chromosome. the cata, catb, and catc genes encode enzymes that catalyze consecutive reactions in the catechol branch of the beta-ketoadipate pathway: cata, catechol-1,2-dioxygenase (ec 1.13.11.1); catb, muconate lactonizing enzyme (ec 5.5.1.1); and catc, muconolactone isomerase (ec 5.3.3.4). a recombinant plasmid, pro1783, which c ...19883139626
cloning, nucleotide sequence and characterization of genes encoding naphthalene dioxygenase of pseudomonas putida strain ncib9816.we have cloned the naphthalene dioxygenase(nd)-coding genes from pseudomonas putida strain ncib9816 based on their ability to convert indole to indigo. the region coding for this enzyme activity was sequenced and three successive open reading frames were found. the corresponding gene products were identified using the t7 polymerase/promoter system. all of them are necessary for the nd activity. a comparison of the nd-coding genes with the ones coding for benzene dioxygenase revealed significant ...19883243438
[purification and properties of beta-ketoadipyl-coenzyme a thiolase from pseudomonas putida]. 19883250096
loss of the toluene-xylene catabolic genes of tol plasmid pww0 during growth of pseudomonas putida on benzoate is due to a selective growth advantage of 'cured' segregants.during growth on benzoate-minimal medium pseudomonas putida mt-2 (paw1) segregates derivative ('cured') strains which have lost the ability to use the pathway encoded by its resident catabolic plasmid pww0. experiments with two plasmids identical to pww0 but each with an insert of tn401, which confers resistance to carbenicillin, suggested that the 'benzoate curing' occurs far more frequently by the specific deletion of the 39 kbp region carrying the catabolic genes than by total plasmid loss. t ...19883246596
specificity of pyoverdine-mediated iron uptake among fluorescent pseudomonas strains.pyoverdine-mediated iron transport was determined for seven fluorescent pseudomonas strains belonging to different species. for all strains, cell or cell outer membrane and iron(iii)-pyoverdine combinations were compared with their homologous counterparts in uptake, binding, and cross-feeding experiments. for four strains (pseudomonas putida atcc 12633, pseudomonas fluorescens w, p. fluorescens atcc 17400, and pseudomonas tolaasii ncppb 2192), the pyoverdine-mediated iron transport appeared to b ...19883170485
physiological comparison of d-cysteine desulfhydrase of escherichia coli with 3-chloro-d-alanine dehydrochlorinase of pseudomonas putida cr 1-1.d-cysteine desulfhydrase of escherichia coli w3110 delta trped102/f' delta trped102 was physiologically characterized. it was found to be located in the cytosolic fraction, as 3-chloro-d-alanine dehydrochlorinase is. d-cysteine desulfhydrase catalyzed not only the alpha, beta-elimination reaction of o-acetyl-d-serine to form pyruvate, acetic acid and ammonia, but also the beta-replacement reaction of o-acetyl-d-serine with sulfide to form d-cysteine. however, these reactions appeared not to proc ...19883132906
[clinical study of cefuzonam in the field of obstetrics and gynecology].cefuzonam (czon, l-105) was used clinically for the treatment of obstetrical and gynecological infections at a dosage of 1 g once or twice daily by intravenous drip infusion. the results obtained are summarized as follows. 1. clinical effects of czon were analyzed in 10 patients, including 5 patients with intrapelvic infections, 3 with intrauterine infections, and 1 each with adnexitis and an external genital infection. excellent responses were observed in 1 patient (11.1%), good responses in 7 ...19883172464
transformation of bacteria with plasmid dna by electroporation.the possibility of electric field-mediated transformation ("electroporation") of a gram-positive bacterium (enterococcus faecalis) and two gram-negative bacteria (escherichia coli and pseudomonas putida) with plasmid dna was investigated. e. faecalis protoplasts could be transformed by electroporation with a transformation frequency of 10(4) to 10(5) transformants/micrograms plasmid. untreated--i.e., washed--cells of e. coli could be transformed with rates of 1 x 10(5) transformants/micrograms p ...19883133958
cloning and expression of pca genes from pseudomonas putida in escherichia coli.beta-ketoadipate elicits expression of five structural pca genes encoding enzymes that catalyse consecutive reactions in the utilization of protocatechuate by pseudomonas putida. three derivatives of p. putida prs2000 were obtained, each carrying a single copy of tn5 dna inserted into a separate region of the genome and preventing expression of different sets of pca genes. selection of tn5 in or near the pca genes in these derivatives was used to clone four structural pca genes and to enable the ...19883076176
physical and functional mapping of two cointegrate plasmids derived from rp4 and tol plasmid pdk1.cointegrate plasmids were formed in vivo between the broad-host-range r-plasmid rp4 and two catabolic plasmids derived from pseudomonas putida hs1. one of these was the wild-type plasmid pdk1 encoding the complete inducible toluene/xylene (tol) catabolic pathway and one was pdkt1, a deletion derivative of pdk1 selected after growth of hs1 on benzoate and supporting growth on only toluene. the two plasmids formed, pdk2 and pdkt2 respectively, each consisted of a complete rp4 replicon in which was ...19883076182
[physical map of the dna of bacteriophage tf of pseudomonas putida].a physical map has been constructed for p. putida bacteriophage tf dna containing single-strand breaks (nicks). localization of cleavage sites for ecori, hindiii, hpai clai, bamhi, sali, xbai and xhoi restriction endonucleases was determined. position of single-strand breaks was mapped by electrophoretic analysis of denatured tf dna and electron microscopy of partially denatured dna samples. the tf genome is characterized by the presence of two classes of nicks differing in the frequency of thei ...19883173374
[purification and properties of two enzymes of meta-cleaving the aromatic ring controlled by the biphenyl biodegradation plasmid pbs 241 from pseudomonas putida].it was shown that two metapyrocatechases (ec 1.13.11.2) function in pseudomonas putida bs893. biphenyl degradative plasmid pbs241 carries the genes of these enzymes. the basic properties of the both enzymes, i. e., mpc1 and mpc2, were investigated. it was found that mpc1 is an enzyme with a molecular mass of 135 kd and has a heterotetrameric subunit structure (alpha 2 beta 2), being made up of two non-identical polypeptides with mr of 34 and 22.5 kd; pi is 5.15, the ph optimum is at 8.0, a tempe ...19883179349
[isolation, purification and various properties of l-lysine-2-monooxygenase from pseudomonas putida].isolation and purification of l-lysine-2- monooxygenase from the bacterium pseudomonas species was carried out. the purification procedure included ammonium sulfate fractionation, acid treatment, gel filtration through sephadex g-200 and ion-exchange chromatography on deae-sephadex a-50. such treatment resulted in more than 220-fold purification and 22% yield; the specific activity of the enzyme is 14.6 u/mg. the enzyme spectrum is typical for flavoproteins, with peaks at 275, 386 and 462 nm. at ...19883191192
4-methoxybenzoate monooxygenase from pseudomonas putida: isolation, biochemical properties, substrate specificity, and reaction mechanisms of the enzyme components. 19883226294
in vivo generation of r68.45-ppgh1 hybrid plasmids conferring a phl+ (meta pathway) phenotype.plasmid ppgh1 originating from pseudomonas putida strain h carries all the genes required for the degradation of phenol (or cresols) via the meta cleavage pathway. besides mobilization of ppgh1 by a plasmid of the incompatibility group p-1, hybrid plasmids conferring the phl+ phenotype could be selected, when r68.45 was the conjugative plasmid. the hybrids contain the complete r68.45 and part of ppgh1. integration of phl-dna of ppgh1 into r68.45 occurred exclusively via the is21 region of r68.45 ...19883226424
properties and functions of two succinic-semialdehyde dehydrogenases from pseudomonas putida.two forms of succinic-semialdehyde dehydrogenase have been isolated in pseudomonas putida. the two enzymes could be separated by filtration on sephacryl s-300 and their apparent molecular weights were approx. 200,000 and 100,000. the smaller enzyme, which is induced by growth on 4-hydroxyphenylacetate, has been purified to 88% homogeneity by anion-exchange and affinity chromatography. electrophoresis in sodium dodecyl sulphate gave rise to a molecular weight of 53,000, indicating that the native ...19883355840
nucleotide sequence of the regulatory gene xylr of the tol plasmid from pseudomonas putida.we have determined the nucleotide sequence of the xylr gene for a transcriptional activator for the degradative pathway of aromatic hydrocarbons on the tol plasmid from pseudomonas putida. the 1698-bp sequence for a 566-amino acid (aa) protein (mr 63741) was identified as the xylr-encoding sequence. three regions in xylr show homology to klebsiella pneumoniae ntrc and nifa, both of which are transcriptional activators for the ntr and nif genes involved in the nitrogen metabolism. the central reg ...19883169574
in vitro and in vivo antibacterial activities of me1207, a new oral cephalosporin.me1207 (pivaloyloxymethyl ester of me1206) is a new oral cephalosporin. me1206 is (6r,7r)-7-[(z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)- acetamido]-3-[(z)-2-(4-methylthiazol-5-yl)-ethyl]-cephem-4-carboxy lic acid. the susceptibilities of about 1,600 clinical isolates to me1206 were determined by the agar dilution method. me1206 showed a broad spectrum of activity against gram-positive and gram-negative bacteria. me1206 was more active than cefaclor, t-2525, and cefixime against staphylococcus ...19883264132
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