PMID(sorted ascending)
chemotaxis of pseudomonas putida toward chlorinated benzoates.the chlorinated aromatic acids 3-chlorobenzoate and 4-chlorobenzoate are chemoattractants for pseudomonas putida prs2000. these compounds are detected by a chromosomally encoded chemotactic response to benzoate which is inducible by beta-ketoadipate, an intermediate of benzoate catabolism. plasmid pac27, encoding enzymes for 3-chlorobenzoate degradation, does not appear to carry genes for chemotaxis toward chlorinated compounds.19902339899
genes for two herbicide-inducible cytochromes p-450 from streptomyces griseolus.streptomyces griseolus atcc 11796 contains two inducible, herbicide-metabolizing cytochromes p-450 previously designated p-450su1 and p-450su2 (p-450cva1 and p-450cvb1, respectively, using nomenclature of nebert et al. [d. w. nebert, m. adesnik, m. j. coon, r. w. estabrook, f. j. gonzalez, f. p. guengerich, i. c. gunsalus, e. f. johnson, b. kemper, w. levin, i. r. phillips, r. sato, and m. r. waterman, dna 6:1-11, 1987]). using antibodies directed against cytochrome p-450su1, its n-terminal amin ...19902345149
purification and characterization of salicylate hydroxylase from pseudomonas putida ppg7.the salicylate hydroxylase from p. putida ppg7 was purified and characterized. the enzyme appears to be monomeric, and it showed one major band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with an apparent mr of 45 kda. the sequence of the first 25 amino acids of salicylate hydroxylase (ppg7) was determined. also, the total amino acid composition of salicylate hydroxylase (ppg7) was obtained and compared with that of the known salicylate hydroxylase from p. putida.19902363715
[the sos-like reactions of pseudomonas putida].under ultraviolet radiation of pseudomonas putida 1087 the sos-similar response which is expressed in the inhibition of cell respiration and cell division with the following filamentation is revealed. in the result of introduction of ppe24 and pmh21 plasmids into the cells of p. putida 1087 for inhibition of reca-similar protein the sos-similar response disappears and the basic cell mass dies.19902372559
[new plasmids of herbicide 2,4-dichlorophenoxyacetic acid biodegradation].three herbicide 2,4-d metabolizing bacterial strains were isolated from three independent soil samples of estonia. the strains, although belonging to various species, contain 2,4-d degradative plasmids with identical restriction patterns. pest4001 is a 78 kb conjugative plasmid. all pseudomonas putida paw340 2,4-d+ transconjugants obtained a 70 kb plasmid pest4011 - a deletion derivative of the pest4001. the restriction patterns of the plasmids mentioned above are considerably different from tho ...19902373362
the downfield resonances in the 1h nmr spectra of azotobacter vinelandii and pseudomonas putida seven-iron ferredoxins.pseudomonas putida and azotobacter vinelandii ferredoxins each contain one [4fe-4s] cluster and one [3fe-4s] cluster. their polypeptide chains are nearly identical, differing by only 15 residues out of a total of 106. t1 measurements and temperature dependence studies of the 1h nmr spectrum of each ferredoxin demonstrate that all six resolved downfield resonances are near an iron-sulfur center. the five most downfield resonances are shown to arise from protons on cysteinyl beta-carbons by incorp ...19902373698
[peripheral metabolism of pseudomonal putida transconjugants degrading chloro- and methylaromatic compounds].peripheral metabolism was studied in the pseudomonas putida 37cc transconjugant. in the strain grown on benzoate, pyrocatechase (pc) i with a low activity to chlorocatechols was induced, whereas pcii actively decomposing chlorocatechols was induced during its growth on 3-chlorobenzoic acid. the p. putida 37cc transconjugant grown on alpha-methylstyrene (ms) exerted the activity of both metapyrocatechase (mpc) and pc, whereas in the parent strain p. putida r-1 only mpc was involved in the degrada ...19902374510
purification and characterization of d-2-haloacid dehalogenase from pseudomonas putida strain aj1/23.a d-2-haloacid dehalogenase was isolated and purified to homogeneity from pseudomonas putida strain aj1/23. the enzyme catalysed the stereospecific dehalogenation of the d-isomer of 2-chloropropionate. using a new ion-chromatograph assay, the enzyme was found to catalyse the dehalogenation of short-chain 2-halocarboxylic acids. maximum enzyme activity occurred at ph 9.5 and 50 degrees c and the enzyme was insensitive to most -sh reagents. the enzyme has an mr of about 135,000 and appears to be c ...19902380688
chromium reduction in pseudomonas putida.reduction of hexavalent chromium (chromate) to less-toxic trivalent chromium was studied by using cell suspensions and cell-free supernatant fluids from pseudomonas putida prs2000. chromate reductase activity was associated with soluble protein and not with the membrane fraction. the crude enzyme activity was heat labile and showed a km of 40 microm cro4(2-). neither sulfate nor nitrate affected chromate reduction either in vitro or with intact cells.19902389940
evolutionary differences in chromosomal locations of four early genes of the tryptophan pathway in fluorescent pseudomonads: dna sequences and characterization of pseudomonas putida trpe and trpgdc.pseudomonas putida possesses seven structural genes for enzymes of the tryptophan pathway. all but one, trpg, which encodes the small (beta) subunit of anthranilate synthase, have been mapped on the circular chromosome. this report describes the cloning and sequencing of p. putida trpe, trpg, trpd, and trpc. in p. putida and pseudomonas aeruginosa, dna sequence analysis as well as growth and enzyme assays of insertionally inactivated strains indicated that trpg is the first gene in a three-gene ...19902404959
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
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
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
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 and muconolactone isomerase (ec, 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 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
purification and characterization of adhesive exopolysaccharides from pseudomonas putida and pseudomonas 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
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
analysis of nonpolar insertion mutations in the trfa gene of incp plasmid rk2 which affect its broad-host-range property.replication of broad-host-range plasmid rk2 requires the protein product(s) of the plasmid-encoded trfa gene to initiate replication at oriv, the vegetative replication origin. the trfa gene contains two translational starts which direct translation of two polypeptides, of 382 and 285 amino acids, which differ by the 97 amino acids at their n-terminus. nonpolar insertions which abolish expression of the larger trfa polypeptide but otherwise retain the trfa gene's normal expression signals severe ...19892495025
selection and characterization of a mutant of the cloned gene for mandelate racemase that confers resistance to an affinity label by greatly enhanced production of enzyme.the plasmid pscr1 containing the gene for mandelate racemase (ec from pseudomonas putida (atcc 12633) allows pseudomonas aeruginosa (atcc 15692) to grow on (r)-mandelate as its sole carbon source [ransom, s. c., gerlt, j. a., powers, v. m., & kenyon, g. l. (1988) biochemistry 27, 540]; the chromosome of the p. aeruginosa host apparently does not contain the gene for mandelate racemase but does contain genes for the remaining enzymes in the mandelate pathway and enables growth on (s)-man ...19892496759
dna sequence of the tryptophan synthase genes of pseudomonas putida.genes encoding the 2 subunits of tryptophan synthase in pseudomonas putida have been identified and cloned by their similarity to the corresponding genes in pseudomonas aeruginosa. the deduced amino acid sequences were confirmed by comparison with regions ascertained earlier by protein sequencing. the pseudomonas amino acid sequences are 85% identical for the beta subunit and 70% identical for the alpha subunit. these sequences are compared to those of salmonella typhimurium, where the structure ...19892503057
plasmid control of the pseudomonas aeruginosa and pseudomonas putida phenotypes and of linalool and p-cymene oxidation.two pseudomonas strains (ppg777 and pag158) were derived from the parent isolate pseudomonas incognita (putida). strain ppg777 resembles the parental culture in growth on linalool as a source of carbon and slight growth on p-cymene, whereas pag158 grows well on p-cymene, but not on linalool or other terpenes tested, and has a p. aeruginosa phenotype. curing studies indicate that linalool metabolism is controlled by an extrachromosomal element whose loss forms a stable strain pag158 with the p-cy ...19892504698
synthesis of poly-3-hydroxyalkanoates is a common feature of fluorescent pseudomonads.the fluorescent pseudomonads are classified as a group, one characteristic of which is that they do not accumulate poly-3-hydroxybutyrate (phb) during nutrient starvation in the presence of excess carbon source. in this paper we show that prototype strains from this subclass, such as pseudomonas aeruginosa, pseudomonas putida, and pseudomonas fluorescens, do accumulate poly-3-hydroxyalkanoates (pha) when grown on fatty acids. these phas are composed of medium-chain-length (c6 to c12) 3-hydroxy f ...19892506811
microbial metabolism of quinoline and related compounds. ii. degradation of quinoline by pseudomonas fluorescens 3, pseudomonas putida 86 and rhodococcus spec. b1.quinoline catabolism was investigated with different bacterial strains, able to use quinoline as sole source of carbon, nitrogen and energy. some degradation products of quinoline were isolated from the culture fluids and identified. with pseudomonas fluorescens and pseudomonas putida we found 2-oxo-1,2-dihydroquinoline, 8-hydroxy-2-oxo-1,2-dihydroquinoline, 8-hydroxycoumarin and 2,3-dihydroxyphenylpropionic acid as intermediates. with a rhodococcus strain 2-oxo-1,2-dihydroquinoline, 6-hydroxy-2 ...19892514722
factors affecting conjugal transfer of plasmids encoding mercury resistance from pure cultures and mixed natural suspensions of epilithic bacteria.sixty-five pure cultures of epilithic bacteria were examined for their ability to transfer mercury resistance to pseudomonas aeruginosa; five isolates transferred plasmids encoding mercury resistance with frequencies ranging from 8.4 x 10(-8) to 2.8 x 10(-3) per recipient. two of the plasmids, pqm3 and pqm4, encoded narrow-spectrum mercury resistance, pqm3 also encoded streptomycin resistance, and both plasmids were broad host range. maximum transfer frequencies of epilithic plasmids from pure c ...19892515247
[expression of the phospholipase c gene of pseudomonas aeruginosa in escherichia coli and pseudomonas].the plc gene for phospholipase of pseudomonas aeruginosa, able to be transcribed only from its own promoter, has been introduced into escherichia coli, pseudomonas aeruginosa and pseudomonas putida cells in the recombinant plasmid ppms21 of a wide host range. the expression of plc gene in all recipient cells has been shown to be phosphate regulated. the fact emphasizes the identity of pho-regulation systems in escherichia coli and pseudomonas cells. the level of phospholipase activity is similar ...19892515430
selective medium for pseudomonas aeruginosa that uses 1,10-phenanthroline as the selective agent.the mic of 1,10-phenanthroline for 35 pseudomonas aeruginosa strains was 128 micrograms/ml, whereas 32 micrograms or less per ml inhibited all other microorganisms tested. on the basis of these results, a selective agar for p. aeruginosa which contained 15 g of trypticase soy broth (bbl microbiology systems), 15 g of agar, and 0.1 g of phenanthroline per liter was formulated. forty-four p. aeruginosa strains yielded a mean efficiency of plating on this medium of 79% of the counts obtained on try ...19892515804
growth of genetically engineered pseudomonas aeruginosa and pseudomonas putida in soil and rhizosphere.the effect of the addition of a recombinant plasmid containing the pgla gene encoding an alpha-1,4-endopolygalacturonase from pseudomonas solanacearum on the growth of pseudomonas aeruginosa and pseudomonas putida in soil and rhizosphere was determined. despite a high level of polygalacturonase production by genetically engineered p. putida and p. aeruginosa, the results suggest that polygalacturonase production had little effect on the growth of these strains in soil or rhizosphere.19892515805
cloning and analysis of genes involved in coenzyme b12 biosynthesis in pseudomonas denitrificans.cobalamin synthesis probably requires 20 to 30 different enzymatic steps. pseudomonas putida and agrobacterium tumefaciens mutants deficient in cobalamin synthesis (cob have been isolated. in p. putida, cob mutants were identified as being unable to use ethanolamine as a source of nitrogen in the absence of added cobalamin (deamination of ethanolamine requires coenzyme b12 as a cofactor). in a. tumefaciens, cob mutants were simply screened for their reduced cobalamin synthesis. a genomic library ...19892536665
cloning and characterization of a gene encoding an outer membrane protein required for siderophore-mediated uptake of fe3+ in pseudomonas putida iron-limited environments plant-growth-stimulating pseudomonas putida wcs358 produces a yellow-green fluorescent siderophore called pseudobactin 358. ferric pseudobactin 358 is efficiently taken up by cells of wcs358 but not by cells of another rhizophere-colonizing strain, pseudomonas fluorescens wcs374. a gene bank containing partial sau3a dna fragments from wcs358 was constructed in a derivative of the broad-host-range cosmid plafr1. by mobilization of this gene bank to strain wcs374 a cos ...19892540157
structure of the dnaa region of pseudomonas putida: conservation among three bacteria, bacillus subtilis, escherichia coli and p. putida.we have cloned from pseudomonas putida a gene homologous to escherichia coli dnaa, and determined the sequence of the gene and its neighboring region. the dnaa gene and at least three other genes, dnan, recf and gyrb, were found to be highly homologous to the genes in the dnaa regions of the e. coli and bacillus subtilis chromosomes. a non-translatable region of some 600 bp immediately upstream of the dnaa gene is also conserved in the three bacteria and contains 3, 12, and 14 dnaa-boxes (ttatcc ...19892540413
omegon-km: a transposable element designed for in vivo insertional mutagenesis and cloning of genes in gram-negative combine the features of the omega interposons with the advantages of in vivo transposition mutagenesis, we have constructed an artificial transposon, called omegon-km. the omegon-km transposon is carried on the plasmid pjff350 which can be conjugally mobilized into a broad range of gram-negative bacteria. omegon-km is flanked, in inverted orientation, by synthetic 28-bp repeats derived from the ends of is1. in addition, each end of omegon-km has the very efficient transcription and translatio ...19892546859
purification and some properties of a 2fe ferredoxin in pseudomonas ovalis.a [2fe-2s] ferredoxin was found in pseudomonas ovalis which was grown in a medium supplemented with glucose and ammonium sulfate. the molecular weight of the 2fe ferredoxin was estimated to be 13,000. it contained 2.2 gramatoms of non-heme iron and 2.3 gramatoms of acid-labile sulfur per mole protein. the absorption and circular dichroism spectra were characteristic of those of [2fe-2s] type ferredoxins, especially adrenodoxin and putidaredoxin. the electron paramagnetic resonance spectrum of th ...19892548508
a simple procedure for transferring genes cloned in escherichia coli vectors into other gram-negative bacteria: phenotypic analysis and mapping of tol plasmid gene xylk.a simple method to transfer non-conjugative escherichia coli plasmids to other gram-negative bacteria and their maintenance is described. this method involves generation of inverse transposition-mediated cointegrates of the non-conjugative e. coli plasmid with a conjugative incw broad-host-range plasmid, r388, carrying tn10. isolation of such cointegrates was readily effected by conjugal transfer from an e. coli donor containing the two plasmids to an e. coli recipient, with selection for transc ...19892548929
characterization of five genes in the upper-pathway operon of tol plasmid pww0 from pseudomonas putida and identification of the gene products.the upper operon of the tol plasmid pww0 of pseudomonas putida encodes a set of enzymes which transform toluene and xylenes to benzoate and toluates. the genetic organization of the operon was characterized by cloning of the upper operon genes into an expression vector and identification of their products in escherichia coli maxicells. this analysis showed that the upper operon contains at least five genes in the order of xylc-xylm-xyla-xylb-xyln. between the promoter of the operon and xylc, the ...19892549010
cloning of pmol28-encoded nickel resistance genes and expression of the genes in alcaligenes eutrophus and pseudomonas spp.the 163-kilobase-pair (kb) plasmid pmol28, which determines inducible resistance to nickel, cobalt, chromate, and mercury salts in its native host alcaligenes eutrophus ch34, was transferred to a derivative of a. eutrophus h16 and subjected to cloning procedures. after tn5 transposon mutagenesis, restriction endonuclease analysis, and dna-dna hybridization, two dna fragments, a 9.5-kb kpni fragment and a 13.5-kb hindiii fragment (hki), were isolated. hki contained ek1, the kpni fragment, as a su ...19892549012
the purification and characterization of 4-ethylphenol methylenehydroxylase, a flavocytochrome from pseudomonas putida jd1.the enzyme 4-ethylphenol methylenehydroxylase was purified from pseudomonas putida jd1 grown on 4-ethylphenol. it is a flavocytochrome c for which the mr was found to be 120,000 by ultracentrifuging and 126,000 by gel filtration. the enzyme consists of two flavoprotein subunits each of mr 50,000 and two cytochrome c subunits each of mr 10,000. the redox potential of the cytochrome is 240 mv. hydroxylation proceeds by dehydrogenation and hydration to give 1-(4'-hydroxyphenyl)ethanol, which is als ...19892556994
transposable elements for efficient manipulation of a wide range of gram-negative bacteria: promoter probes and vectors for foreign genes.we describe here the construction and use of a series of modified transposons based on the insertion sequence is1. like their parent, omegon-km [fellay et al., gene 76 (1989) 215-226], these elements permit efficient insertional mutagenesis of a variety of gram-negative bacteria. the presence of a functional pbr322 origin of replication within the transposable element facilitates subsequent cloning of the mutated gene. the omegon-km system was previously shown to function in pseudomonas putida, ...19892559879
[cloning and gene expression determining phenol breakdown in pseudomonas putida strains]. 19892561419
study of the 5-oxoprolinase reaction by 13c nmr.5-oxoprolinase catalyzes the atp-dependent decyclization of 5-oxo-l-proline to l-glutamate. previous studies provided evidence for the intermediate formation of a phosphorylated form of 5-oxoproline (seddon, a. p., and meister, a. (1986) j. biol. chem. 261, 11538-11541) and of a tetrahedral intermediate (li, l., seddon, a. p., and meister, a. (1987) j. biol. chem. 262, 11020-11025). a new approach to the study of the reaction mechanism using the 18o isotope effect on the 13c nmr signals for 5-ox ...19892563377
thermal activation of photoactivatable urocanase from pseudomonas putida.the dark inactivation of urocanase from pseudomonas putida is caused by the formation of a sulfite adduct of the tightly bound coenzyme, nicotinamide adenine dinucleotide. photodissociation of this adduct by uv radiation restores the enzyme activity. based on cold exhaustive dialysis the modification reaction appeared to be irreversible. however, we now report that sulfite modification of urocanase is reversible at higher temperatures. an arrhenius plot of the thermal activation is linear (20-38 ...19892570140
the stoichiometry of the tightly bound nad+ in urocanase. separation and characterization of fully active and inhibited forms of the enzyme.1. urocanase, purified by classical methods [keul, v., kaeppeli, f., ghosh, c., krebs, t., robinson, j. a. and rétey, j. (1979) j. biol. chem. 254, 843-851] from pseudomonas putida was submitted to high-performance liquid chromatography on a tsk-deae column. the enzyme was eluted in three resolved peaks (a, b and c) exhibiting specific activities of 3.4 u/mg, 1.85 u/mg and 0.4 u/mg, respectively. 2. the difference spectra of peaks b and a as well as of c and a showed maxima at 330 nm. 3. irradia ...19892574107
specific inhibition of bacterial and bovine urocanases by glycylglycine.urocanase (ec purified from pseudomonas putida was unexpectedly inhibited by the dipeptide glycylglycine. using a spectrophotometric assay for urocanase activity, we characterized the inhibition. the inhibition was temperature-, concentration-, and time-dependent; 0.1, 0.5 and 1.0 mm glycylglycine inhibited the enzyme by 20%, 50% and 78%, respectively, in 60 min at 30 degrees c. dithiothreitol and reduced glutathione did not prevent the process. the inhibition was a pseudo first-order ...19892577699
operon structure and nucleotide homology of the chlorocatechol oxidation genes of plasmids pjp4 and pac27.alcaligenes eutrophus harboring plasmid pjp4 (strain jmp134) is capable of growing on both 2,4-dichlorophenoxyacetate (2,4-d) and 3-chlorobenzoate (3-cba), while pseudomonas putida carrying plasmid pac27 (strain ac867) can utilize only 3-cba as the sole carbon source. the tfdcdef operon of the pjp4 plasmid and the clcabd operon of plasmid pac27 each encode enzymes for the degradation of chlorocatechols (clc), key intermediates in the catabolism of 2,4-d and 3-cba. similarities in the nucleotide ...19892583528
survival of and plasmid stability in pseudomonas and klebsiella spp. introduced into agricultural drainage water.cell survival and plasmid stability in pseudomonas fluorescens r2f and pseudomonas putida cym 318 containing respectively, plasmid rp4 and prk2501, and klebsiella aerogenes nctc 418 harboring plasmid pbr322 were studied in sterile and nonsterile agricultural drainage water under both aerobic and anaerobic conditions and in the absence and presence of added nutrients. both pseudomonas strains survived well in sterile drainage water incubated aerobically, with or without added nutrients. however, ...19892590305
xyle functions as an efficient reporter gene in streptomyces spp.: use for the study of galp1, a catabolite-controlled promoter.we describe the development of a convenient and sensitive reporter gene system for streptomyces spp. based on the use of a promoterless copy of the xyle gene of pseudomonas putida. the xyle gene product is a catechol dioxygenase, which converts the colorless substrate catechol to an intensely yellow hydroxymuconic semialdehyde. a promoterless copy of xyle was placed under the transcriptional control of galp1, a glucose-repressed and galactose-induced promoter from streptomyces lividans, and its ...19892592344
molecular cloning, coding nucleotides and the deduced amino acid sequence of p-450bm-1 from bacillus megaterium.the gene encoding barbiturate-inducible cytochrome p-450bm-1 from bacillus megaterium atcc 14581 has been cloned and sequenced. an open reading frame in the 1.9 kb of cloned dna correctly predicted the nh2-terminal sequence of p-450bm-1 previously determined by protein sequencing, and, in toto, predicted a polypeptide of 410 amino acid residues with an mr of 47,439. the sequence is most, but less than 27%, similar to that of p-450cam from pseudomonas putida, so that p-450bm-1 clearly belongs to ...19892597681
[study of morphology and genome structure of pseudomonas putida bacteriophages for their classification].a group of 27 bacteriophages specific for pseudomonas putida strains ppg1 and ppn has been isolated. the phages were characterized and compared with the previously described virulent (pf 16, af, tf and pmw) and temperate (pp56 and pp71) phages. the new phages belong to b1 and c1 morphotypes, according to ackerman's classification. phage dnas were digested with several endonucleases; the molecular weights and homology of the dnas were determined. all phages of p. putida isolated up to now were di ...19892599372
nucleotide and deduced amino acid sequence of the rpon sigma-factor of pseudomonas putida. 19892602128
survival of pseudomonas putida uwc1 containing cloned catabolic genes in a model activated-sludge unit.the possibility of the accidental or deliberate release of genetically engineered microorganisms into the environment has accentuated the need to study their survival in, and effect on, natural habitats. in this study, pseudomonas putida uwc1 harboring a non-self-transmissible plasmid, pd10, encoding the breakdown of 3-chlorobenzoate was shown to survive in a fully functioning laboratory-scale activated-sludge unit (asu) for more than 8 weeks. the asu maintained a healthy, diverse protozoal popu ...19892604401
monohydroxylation of phenol and 2,5-dichlorophenol by toluene dioxygenase in pseudomonas putida f1.pseudomonas putida f1 contains a multicomponent enzyme system, toluene dioxygenase, that converts toluene and a variety of substituted benzenes to cis-dihydrodiols by the addition of one molecule of molecular oxygen. toluene-grown cells of p. putida f1 also catalyze the monohydroxylation of phenols to the corresponding catechols by an unknown mechanism. respirometric studies with washed cells revealed similar enzyme induction patterns in cells grown on toluene or phenol. induction of toluene dio ...19892604403
cloning and nucleotide sequences of nadh-putidaredoxin reductase gene (cama) and putidaredoxin gene (camb) involved in cytochrome p-450cam hydroxylase of pseudomonas putida.pseudomonas putida ppgl, which carries the cam plasmid encoding enzymes involved in the degradation pathway of d-camphor, can utilize d-camphor as a sole carbon source. cytochrome p-450cam and related enzymes participate in the early oxidation steps of d-camphor degradation metabolism. we cloned from a hindiii dna library of ppgl a 2.9 kbp cam segment which carries the major part of cama gene encoding nadh-putidaredoxin reductase and the entire camb gene encoding putidaredoxin. the 2.9 kbp cam s ...19892613690
[plasmids for biphenyl, chlorobiphenyl and metatoluylate degradation from pseudomonas putida].pseudomonas putida strain su83, harbors the pbs311 plasmid coding for the degradation of biphenyl, 2- and 4-chlorbiphenyl, meta- and paratoluylate. the insertional mutants of the plasmid obtained by the transposon tn5 insertion were isolated. one of the mutants was used for cloning of the biphenyl degradation genes. the plasmid pbs311:: tn5 dna was inserted into the bamhi site of the plasmid pbr322 and cloned. 11 recombinants of 354 tested were treated with 0.1% solution of 2,3-dioxybiphenyl. on ...19892628753
[genetic determination of degradation of ampholytic surfactants].plasmid dna was detected in pseudomonas putida 141 and p. stutzeri at strains which caused destruction of the ampholytic surfactants alkylamino-bis-propionate (aabp) and amidobetaine, respectively. as was demonstrated using genetic analytic procedures, the plasmids controlled aabp and amidobetaine destruction. no plasmid dna was found in p. desmolytica c37 which caused cyclimide destruction or in pseudomonas sp. 1 and citrobacter freundii to strains responsible for aabp destruction. apparently, ...19892636974
extracellular product of nocardia amarae induces bacterial cell flocculation.the fact that nocardia amarae yk1 produced a bacterial flocculation-inducing substance (designated as fix) was discovered. fix had a function of flocculating proliferous cells. fix-induced flocculation was inhibited by making cells resting, but not completely by adding chloramphenicol. fix worked widely on gram-positive to -negative bacteria. in the presence of fix, achromobacter cycloclastus iam1013, acinetobacter calcoaceticus iam1517, bacillus subtilis iam1069, escherichia coli c600-1, e. col ...19892653957
in vivo enzymology: a deuterium nmr study of formaldehyde dismutase in pseudomonas putida f61a and staphylococcus aureus.high-resolution deuterium nmr spectroscopy has been used to follow the detoxifying metabolism of [d2]formaldehyde in vivo in several bacterial species. production of [d2]methanol in escherichia coli confirms that the oxidation and reduction pathways of metabolism are independent in this organism. efficient production of equimolar quantities of [d]formate and [d3]methanol in pseudomonas putida f61a and staphylococcus aureus implicates a formaldehyde dismutase, or "cannizzarase", activity. these o ...19892655705
cloning and expression in escherichia coli of the toluene dioxygenase gene from pseudomonas putida ncib11767.the genes encoding toluene dioxygenase, toluene cis-glycol dehydrogenase and catechol 2.3-oxygenase from pseudomonas putida ncib 11767 were cloned and expressed in escherichia coli hb101 on a 20 kb fragment. the recombinant strain produced indigo and a variety of other coloured products. although the enzymes were expressed in the absence of inducers, further induction was observed in the presence of toluene or benzene, implying the presence of regulatory elements on the 20 kb insert.19892656389
5-carboxymethyl-2-hydroxymuconic semialdehyde dehydrogenases of escherichia coli c and klebsiella pneumoniae m5a1 show very high n-terminal sequence homology.5-carboxymethyl-2-hydroxymuconic semialdehyde (chms) dehydrogenase from escherichia coli c and klebsiella pneumoniae m5a1 have been purified and some of their properties studied. the apparent km values for nad and chms were 11.7 +/- 1.5 microm and 5.2 +/- 1.9 microm, respectively, for the k. pneumoniae enzyme, and 19.5 +/- 2.7 microm and 9.2 +/- 1.4 microm, respectively, for the e. coli enzyme. both enzymes were optimally active at ph 7.5 in sodium phosphate buffer. they had subunit molecular we ...19892656390
[cloning of pseudomonas putida genes responsible for the primary stages of oxidation of naphthalene in escherichia coli cells].data on cloning pseudomonas putida d-plasmid pbs286 (incp-9) genes which are responsible for primary stages of naphthalene oxidation as well as data on the expression of these genes in escherichia coli cells are presented. recombinant plasmid pbs959 containing the whole constitutive naha locus encoding naphthalene dioxygenase, a key enzyme of the naphthalene oxidation pathway, has been constructed on the basis of the puc19 vector. an evidence has been obtained that at least a portion of the sequ ...19892661326
cloning of a carbofuran hydrolase gene from achromobacter sp. strain wm111 and its expression in gram-negative bacteria.a 14-kilobase-pair (kbp) ecori dna fragment that encodes an enzyme capable of rapid hydrolysis of n-methylcarbamate insecticides (carbofuran hydrolase) was cloned from carbofuran-degrading achromobacter sp. strain wm111. when used to probe southern blots containing plasmid and total dnas from wm111, this 14-kbp fragment hybridized strongly to a 14-kbp ecori fragment from the greater than 100-kbp plasmid harbored by this strain but weakly to ecori-digested total dna from achromobacter sp. strain ...19892661544
2-oxoaldehyde metabolism in microorganisms.the properties of methylglyoxal-metabolizing enzymes in prokaryotic and eukaryotic microorganisms were studied systematically and compared with those of mammalian enzymes. the enzymes constitute a glycolytic bypass and convert methylglyoxal into pyruvate via lactate. the first step in this conversion is catalyzed by glyoxalase i, methylglyoxal reductase, or methylglyoxal dehydrogenase. the regulation of the yeast glyoxalase system was analyzed. the system was closely related to the proliferative ...19892663129
[susceptibilities of clinical isolates to antibacterial agents. a study mainly focused on ofloxacin (the second report). reported by the research group for testing ofloxacin susceptibility on clinical isolates].susceptibilities of various clinical isolates to ofloxacin (oflx) and other antibacterial drugs were examined at 128 hospital laboratories in 36 prefectures throughout japan between april, 1986 and march, 1987. the results were totalized with an emphasis mainly on oflx and were compared with data obtained in the previous year. in this study, identification and susceptibility tests of the isolates were carried out at each hospital laboratory and the tests were performed according to the 1-dilutio ...19892664255
identification of multiple repressor recognition sites in the hut system of pseudomonas putida.the hutc gene in pseudomonas putida encodes a repressor protein that negatively regulates the expression of all hut genes. we have overexpressed this cloned hutc gene in escherichia coli to identify p. putida hut regions that could specifically bind the repressor. ten restriction fragments, some of which were partially overlapping and spanned the coding portions of the p. putida hut region, were labeled and tested for their ability to recognize repressor in a filter binding assay. this procedure ...19892666390
involvement of pseudomonas putida rpon sigma factor in regulation of various metabolic functions.the rpon protein was originally identified in escherichia coli as a sigma (sigma) factor essential for the expression of nitrogen regulons. in the present study we cloned the pseudomonas putida rpon gene and identified its gene product as a protein with an apparent molecular weight of 78,000. a mutant rpon gene was constructed by in vitro insertion mutagenesis with a kanamycin cassette. a p. putida rpon mutant was then isolated by replacement of the intact chromosomal rpon gene by the mutant rpo ...19892666396
differences between the manganese- and the iron-containing superoxide dismutases of escherichia coli detected through sedimentation equilibrium, hydrodynamic, and spectroscopic studies.the genome of escherichia coli codes for two superoxide dismutases that may contain either iron (fesod) or manganese (mnsod) at the active site. the crystal structures of mnsods from two bacterial sources (but not e. coli) have been completed, and structural comparisons with the crystal structure of the fesod from either e. coli or pseudomonas ovalis have been made. despite the low degree (less than 50%) of sequence homology between the e. coli enzymes, the two proteins are suggested to be struc ...19892669953
transcription initiation at multiple promoters of the pfl gene by e sigma 70-dependent transcription in vitro and heterologous expression in pseudomonas putida in vitro transcription experiments were used to provide further evidence that the gene encoding pyruvate formate-lyase (ec from escherichia coli is transcribed from seven promoters which cover a region of 1.2 kilobase pairs of dna (g. sawers and a. böck, j. bacteriol., 171:2485-2498, 1989). the results demonstrated that all promoters were recognized by the major rna polymerase holoenzyme species e sigma 70 in vitro. further corroboration for multiple functional promoters came from hete ...19892670899
toluene degradation by pseudomonas putida f1. nucleotide sequence of the todc1c2bade genes and their expression in escherichia coli.the nucleotide sequence of the todc1c2bade genes which encode the first three enzymes in the catabolism of toluene by pseudomonas putida f1 was determined. the genes encode the three components of the toluene dioxygenase enzyme system: reductasetol (toda), ferredoxintol (todb), and the two subunits of the terminal dioxygenase (todc1c2); (+)-cis-(1s, 2r)-dihydroxy-3-methylcyclohexa-3,5-diene dehydrogenase (todd); and 3-methylcatechol 2,3-dioxygenase (tode). knowledge of the nucleotide sequence of ...19892670929
bacterial aromatic ring-cleavage enzymes are classified into two different gene families.dioxygenases that catalyze the cleavage of the aromatic ring are classified into two groups according to their mode of ring fission. substrates of ring-cleavage dioxygenases usually contain hydroxyl groups on adjacent aromatic carbons, and intradiol enzymes cleave the ring between these two hydroxyl groups. extradiol enzymes in contrast cleave the ring between one hydroxylated carbon and its adjacent nonhydroxylated carbon. in this study, we determined the complete nucleotide sequence of nahc, t ...19892670937
physically associated enzymes produce and metabolize 2-hydroxy-2,4-dienoate, a chemically unstable intermediate formed in catechol metabolism via meta cleavage in pseudomonas putida.the meta-cleavage pathway of catechol is a major mechanism for degradation of aromatic compounds. in this pathway, the aromatic ring of catechol is cleaved by catechol 2,3-dioxygenase and its product, 2-hydroxymuconic semialdehyde, is further metabolized by either a hydrolytic or dehydrogenative route. in the dehydrogenative route, 2-hydroxymuconic semialdehyde is oxidized to the enol form of 4-oxalocrotonate by a dehydrogenase and then further metabolized to acetaldehyde and pyruvate by the act ...19892681159
isolation and characterization of altered plasmids in mutant strains of pseudomonas putida ncib 9816.the ability of p. putida ncib 9816 to grow with naphthalene (nah+) and salicylate (sal+) is correlated with the presence of an 83 kilobase (kb) conjugative plasmid, pdtg1. derivatives of pdtg1 were obtained from cells after exposure to halogenated analogs of naphthalene or salicylate. the selection of mutants having a nah-sal- or a nah-sal+ phenotype could be enhanced by the addition of triphenyltetrazolium chloride to the indicator medium. structurally modified plasmids were characterized by re ...19892684156
putidaredoxin competitively inhibits cytochrome b5-cytochrome p-450cam association: a proposed molecular model for a cytochrome p-450cam electron-transfer complex.cytochrome b5 has been genetically engineered to afford a fluorescent derivative capable of monitoring its association with cytochrome p-450cam from pseudomonas putida [stayton, p. s., fisher, m. t., & sligar, s. g. (1988) j. biol. chem. 263, 13544-13548]. in the mutant cytochrome b5, threonine is replaced by a cysteine at position 65 (t65c) and has been labeled with the environmentally sensitive fluorophore acrylodan. in this paper, the physiological p-450cam reductant putidaredoxin, an fe2s2.c ...19892690937
trichloroethylene degradation by escherichia coli containing the cloned pseudomonas putida f1 toluene dioxygenase genes.toluene dioxygenase from pseudomonas putida f1 has been implicated as an enzyme capable of degrading trichloroethylene. this has now been confirmed with escherichia coli jm109(pdtg601) that contains the structural genes (todc1c2ba) of toluene dioxygenase under the control of the tac promoter. the extent of trichloroethylene degradation by the recombinant organism depended on the cell concentration and the concentration of trichloroethylene. a linear rate of trichloroethylene degradation was obse ...19892694960
microbial enzymes for creatinine assay: a review.a novel metabolic pathway for the degradation of creatinine with n-methylhydantoin, n-carbamoylsarcosine and sarcosine as successive intermediates was found to operate in pseudomonas putida 77 and many other microorganisms. enzymes involved in this pathway were purified from cells of p. putida 77 and characterized. the first step, deimination of creatinine, is catalyzed by cytosine deaminase/creatinine deiminase. the following two steps, ring-opening of n-methylhydantoin and decarbamoylation of ...19892695273
cloning and sequence analysis of the ntra (rpon) gene of pseudomonas putida.the gene encoding a sigma factor ntra (rpon) was cloned from pseudomonas putida by cross-hybridization with a probe containing a part of the corresponding escherichia coli gene. the cloned gene complemented an ntra mutation of e. coli in activation of xyl genes on the tol plasmid. the predicted amino acid (aa) sequence of p. putida ntra (497 aa; mr 56,215) is highly homologous to ntra proteins from azotobacter vinelandii (81.7%), klebsiella pneumoniae (52.6%), and rhizobium meliloti (36.1%). the ...19892695395
mutations in genes downstream of the rpon gene (encoding sigma 54) of klebsiella pneumoniae affect expression from sigma 54-dependent promoters.two open reading frames (orfs), designated orf95 and orf162, downstream of the klebsiella pneumoniae sigma 54 structural gene (rpon) have been sequenced and shown to encode polypeptides of 12 kd and 16 kd, respectively. orfs homologous to orf95 are present downstream of four out of five rpon genes sequenced to date from a range of gram-negative bacteria, and orf162 is also conserved, at least in pseudomonas putida. chromosomal mutations have been created in each gene using a kan cassette and bot ...19892695747
evidence that the transcription activator encoded by the pseudomonas putida nahr gene is evolutionarily related to the transcription activators encoded by the rhizobium nodd genes.the nahr gene of the 83-kilobase naphthalene degradation plasmid nah7 of pseudomonas putida encodes a 34-kilodalton polypeptide which binds to the nah and sal promoters to activate transcription of the degradation genes in response to the inducer salicylate. the dna sequence of the nahr gene was determined, and a derived amino acid sequence of the nahr protein was obtained. a computer search for homologous proteins showed that within the first 124 amino-terminal residues, nahr has approximately ...19892703465
construction and nucleotide sequence of a cdna encoding the full-length preprotein for human branched chain acyltransferase.a cdna (1.6 kilobases) for branched chain acyltransferase (e2b) isolated from a human liver library encoded only the amino-terminal half of the protein (hummel, k. b., litwer, s., bradford, a. p., aitken, a., danner, d. j., and yeaman, s. j. (1988) j. biol. chem. 263, 6165-6168). here we report the isolation of other cdnas which encode the carboxyl-terminal half of e2b and the construction of a cdna which encodes the entire pre-e2b. cdna from the original clone encoding the leader sequence, lipo ...19892708389
nad-linked, gsh- and factor-independent aldehyde dehydrogenase of the methylotrophic bacterium, hyphomicrobium x.cell-free extracts of hyphomicrobium x showed nad-dependent aldehyde dehydrogenase activity, provided that nad addition preceded that of aldehyde. activity was lost rather rapidly, especially during purification attempts, but this could be partially masked by including a time-dependent restoration step with thiol compounds in the protocol. the nature of the assay buffer appeared to be critical and stimulation occurred on incorporation of k+ ions in the mixture. an even higher specific activity c ...19892712573
identification of pseudomonas alcaligenes chromosomal dna in the plasmid dna of the chlorobenzene-degrading recombinant pseudomonas putida strain cb1-9.the recombinant pseudomonas putida strain cb1-9, which acquired the ability to grow on chlorobenzenes, contains a 33-kilobase (kb) plasmid (pkfl3) which lacked homology to an indigenous 15-kb plasmid (pkfl1) in pseudomonas alcaligenes c-0 parent but was homologous to a 55-kb plasmid (pkfl2) from the p. putida r5-3 parent. chromosomal dna of p. alcaligenes c-0 hybridized to probes prepared from pkfl3 but not to probes prepared from pkfl2. a single clone from a genomic library of p. alcaligenes c- ...19892729978
cloning of bacterial genes specifying degradation of 4-chlorobiphenyl from pseudomonas putida ou83.genes capable of 4-chlorobiphenyl (4-cbp) degradation were cloned from 4-cbp-degrading pseudomonas putida ou83 by using a genomic library which was constructed in the broad-host-range cosmid vector pcp13. p. putida ac812 containing chimeric cosmid-expressing enzymes involved in the 4-cbp degradation pathway were identified by detecting 3-phenylcatechol dioxygenase activity (3-pda). chimeric cosmid clones poh83, poh84, poh85, poh87, and poh88 positive for 3-pda grew in synthetic basal medium cont ...19892729981
[production of antibiotic substances by natural variants of the marine bacterium vibrio fischeri].it was shown that under definite conditions there was competition between natural variants of sea bacteria belonging to v. fischeri. natural variants of v. fischeri, strain 6 differed in their resistance to streptomycin and had different growth rates under conditions of limited aeration. morphologically all the variants were identical. v. fischeri p-0, v. fischeri p-1 and v. fischeri p-2 were studied. the study revealed that v. fischeri p-0 produced a non-dialysing thermostable trypsin-sensitive ...19892730220
flagellation of pseudomonas putida and analysis of its motile behavior.pseudomonas putida flagella were examined. also, changes in motile behavior in response to chemoattractants were analyzed quantitatively by computer. reversals in the rotation direction of bundles of polar flagella resulted in changes in swimming direction. cells swimming in buffer changed direction once every 2 s on average, whereas cells exposed to the attractant benzoate changed direction an average of once every 10 s. the findings show that p. putida responds to temporal gradients of chemoat ...19892738028
[nucleotide sequence of the rpll gene coding for ribosomal protein l7/l12 of pseudomonas putida].the pseudomonas putida rpl l gene coding for ribosomal protein l7/l12 was cloned and sequenced. although asp55 residue in l7/l12 was previously shown to be conservative in ten different organisms, the pseudomonas putida l7/l12 proved to contain asn55, thus showing that asp55 is not invariant.19892751714
a methyl-accepting protein is involved in benzoate taxis in pseudomonas putida.pseudomonas putida is attracted to at least two groups of aromatic acids: a benzoate group and a benzoylformate group. members of the benzoate group of chemoattractants stimulated the methylation of a p. putida polypeptide with an apparent molecular weight of 60,000 in sodium dodecyl sulfate-polyacrylamide gels. this polypeptide is presumed to be a methyl-accepting chemotaxis protein for several reasons: its molecular weight is similar to the molecular weights of escherichia coli methyl-acceptin ...19892768186
degradation of phenol and m-toluate in pseudomonas sp. strain est1001 and its pseudomonas putida transconjugants is determined by a multiplasmid system.the utilization of phenol, m-toluate, and salicylate (phe+, mtol+, and sal+ characters, respectively) in pseudomonas sp. strain est1001 is determined by the coordinated expression of genes placed in different plasmids, i.e., by a multiplasmid system. the natural multiplasmid strain est1001 is phenotypically unstable. in its phe-, mtol-, and sal- segregants, the plasmid dna underwent structural rearrangements without a marked loss of plasmid dna, and the majority of segregants gave revertants. th ...19892768199
degradation of 2-bromo-, 2-chloro- and 2-fluorobenzoate by pseudomonas putida clb 250.pseudomonas putida strain clb 250 (dsm 5232) utilized 2-bromo-, 2-chloro- and 2-fluorobenzoate as sole source of carbon and energy. degradation is suggested to be initiated by a dioxygenase liberating halide in the first catabolic step. after decarboxylation and rearomatization catechol is produced as a central metabolite which is degraded via the ortho-pathway. after inhibition of ring cleavage activities with 3-chlorocatechol, 2-chlorobenzoate was transformed to catechol in nearly stoichiometr ...19892777062
identification of nucleotides critical for activity of the pseudomonas putida catbc promoter.pseudomonas putida utilizes the catbc operon, which encodes cis,cis-muconate lactonizing enzyme i (mlei; ec and muconolactone isomerase (mi; ec, for growth on benzoate as a sole carbon source. this operon is positively regulated, and the promoter is located 64 bp upstream of the catb translational start site. using site-specific mutagenesis, we identified nucleotides that influenced the induction of this promoter. promoter activity was monitored with the promoter probe vector p ...19892779516
cloning, expression, and regulation of the pseudomonas cepacia protocatechuate 3,4-dioxygenase genes.the genes for the alpha and beta subunits of the enzyme protocatechuate 3,4-dioxygenase (ec were cloned from the pseudomonas cepacia dbo1 chromosome on a 9.5-kilobase-pair psti fragment into the broad-host-range cloning vector pro2317. the resultant clone was able to complement protocatechuate 3,4-dioxugenase mutations in p. cepacia, pseudomonas aeruginosa, and pseudomonas putida. expression studies showed that the genes were constitutively expressed and subject to catabolite repressi ...19892808302
genetic organization and sequence of the pseudomonas cepacia genes for the alpha and beta subunits of protocatechuate 3,4-dioxygenase.the locations of the genes for the alpha and beta subunits of protocatechuate 3,4-dioxygenase (ec on a 9.5-kilobase-pair psti fragment cloned from the pseudomonas cepacia dbo1 chromosome were determined. this was accomplished through the construction of several subclones into the broad-host-range cloning vectors pro2317, pro2320, and pro2321. the ability of each subclone to complement mutations in protocatechuate 3,4-dioxygenase (pcaa) was tested in mutant strains derived from p. cepa ...19892808303
[mutants of the plasmid for biodegradation of naphthalene, determining catechol oxidation via the meta-pathway].most of the known naphthalene biodegradation plasmids determine the process of naphthalene degradation via salicylate and catechol using the meta pathway of catechol degradation. however, pseudomonas putida strains with plasmids pbs2, pbs216, pbs217 and npl-1 exert no activity of the enzymes involved in the meta pathway of catechol degradation. when 2-methylnaphthalene was added to the medium as a sole carbon source, mutants growing on this compound were isolated in the strains with the studied ...19892811710
[preservation of the viability of opisthorchis eggs by joint cultivation with pseudomonas putida].the effect of pseudomonas putida on opisthorchis' ova was studied with a view to assess the feasibility of using bacteria as biological agents against opisthorchiasis. experiments on mixed culture of the above-mentioned bacteria and helminths' ova demonstrated the lack of ovicidal effect of pseudomonas putida on the ova.19892811755
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
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
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
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
[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
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