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microbial metabolism of quinoline and related compounds. v. degradation of 1h-4-oxoquinoline by pseudomonas putida 33/1.a bacterial strain was isolated with the ability to use 1h-4-oxoquinoline as the sole source of carbon, nitrogen and energy. on the basis of its physiological properties, this isolate was classified as pseudomonas putida. 1h-3-hydroxy-4-oxoquinoline, n-formylanthranilic acid, anthranilic acid and catechol were identified as intermediates in the degradation pathway. the latter was further degraded by ortho-cleavage. the enzymatic conversion of 1h-4-oxoquinoline into 1h-3-hydroxy-4-oxoquinoline re ...19901963786
mechanism of action of urocanase. specific 13c-labelling of the prosthetic nad+ and revision of the structure of its adduct with imidazolylpropionate.1. [4-13c]nicotinate was synthesised and used to support the growth of a nicotinate auxotrophic mutant of pseudomonas putida. 13c-nmr spectroscopy of the isolated urocanase confirmed the efficient incorporation of 13c into c4 of the nicotinamide ring of the tightly bound nad+ cofactor. 2. beta-[( 2'-13c]imidazol-4-yl)propionate was synthesised according to known procedures and used for inhibition of the 13c-labelled urocanase. an increase in the absorbance at 330 nm indicated adduct formation be ...19901976515
nucleotide sequencing and characterization of pseudomonas putida catr: a positive regulator of the catbc operon is a member of the lysr family.pseudomonas putida utilizes the catbc operon for growth on benzoate as a sole carbon source. this operon is positively regulated by the catr protein, which is encoded from a gene divergently oriented from the catbc operon. the catr gene encodes a 32.2-kilodalton polypeptide that binds to the catbc promoter region in the presence or absence of the inducer cis-cis-muconate, as shown by gel retardation studies. however, the inducer is required for transcriptional activation of the catbc operon. the ...19901688844
molecular analysis of the hydrogenosomal ferredoxin of the anaerobic protist trichomonas vaginalis.we have determined the primary structure of the [2fe-2s]ferredoxin of the anaerobic protist trichomonas vaginalis. this protein, situated in the hydrogenosome, is composed of 93 amino acids. a comparison of t. vaginalis ferredoxin with greater than 80 other ferredoxins shows the closest similarity to [2fe-2s]putidaredoxin of the aerobic bacterium pseudomonas putida and a lesser one to mitochondrial [2fe-2s]ferredoxins of vertebrates. this similarity is reflected in the overall primary structure ...19901696716
stereochemical aspects of the oxidation of 4-ethylphenol by the bacterial enzyme 4-ethylphenol methylenehydroxylase.the o2-independent hydroxylase 4-ethylphenol methylenehydroxylase (4epmh) from pseudomonas putida jd1 catalysed the complete conversion of 4-ethylphenol into 1-(4-hydroxyphenyl)ethanol together with a small amount of 4-hydroxyacetophenone, but with no formation of the side product 4-vinylphenol reported to be formed when the similar enzyme p-cresol methylhydroxylase (pcmh) catalyses this reaction. the enantiomer of 1-(4-hydroxyphenyl)ethanol produced by 4epmh was r(+) when horse heart cytochrome ...19901697166
microbial degradation of the morphine alkaloids: identification of morphine as an intermediate in the metabolism of morphine by pseudomonas putida m10.a strain of pseudomonas putida was isolated by selective enrichment with morphine that was capable of utilising morphine as a primary source of carbon and energy for growth. experiments with whole cells showed that both morphine and codeine, but not thebaine, could be utilised. a novel nadp-dependent dehydrogenase, morphine dehydrogenase, was purified from crude cell extracts and was shown to be capable of oxidising morphine and codeine to morphinone and codeinone, respectively. this nadp-depend ...19901701625
[pseudomonas aps nutrient medium for the isolation and identification of pseudomonas aeruginosa and pseudomonas putida].pseudomonas aps selective medium has been developed on the basis of a newly detected selective antibacterial action of oxaphenamide (p-oxyphenylsalicylamide), a cholagogue. this medium permits a single-stage combined isolation and identification of p. aeruginosa after 16-24 hrs incubation of inoculated material at 42 degrees c. if the material is incubated at 35-37 degrees c, isolation of p. putida and p. aeruginosa is possible, that are differentiated by a nitroreductase microtest within 3 hrs. ...19901705609
molecular cloning of the xyll-xyle region from the p. putida tol plasmid, pdk1.a 5.2 kilobase ecori restriction fragment from the pseudomonas putida hs1 tol plasmid pdk1, encoding a portion of the lower toluene degradation pathway, was cloned into the e. coli plasmid pbr325. a detailed map of the restriction endonuclease sites was constructed and the nucleotide sequence of three contiguous xhoi fragments, with a combined total length of approximately 3.9 kilobases, has been investigated. this region was determined to contain a total of four separate open reading frames, ea ...19901366507
mechanism of formaldehyde biodegradation by pseudomonas putida.formaldehyde biodegradation by a strain of pseudomonas putida has been studied. the results indicate that this biodegradation is initiated by a dismutation reaction, yielding as products formic acid and methanol. the degradation of methanol and formic acid begins after exhaustion of formaldehyde in the medium, and presents a diauxic pattern: first formic acid is consumed followed by methanol. moreover, cell viability, which is affected by the amount of added formaldehyde, has been determined.19901366532
immobilized and free cell continuous cultures of a recombinant e. coli producing catechol 2,3-dioxygenase in a two-stage chemostat: improvement of plasmid stability.the immobilization of recombinant strains of e. coli w3110/ptg205 in k-carrageenan gel beads improves the plasmid stability during continuous cultures in the absence of selection pressure. since, xyl e gene (which encodes catechol 2,3-dioxygenase from pseudomonas putida) transcription is controlled by the trp promoter, the effects of tryptophan (repressor) and 3 beta-indolyl acrylic acid (derepressor) on ptg 205 stability and enzyme production have been studied in both free and immobilized cell ...19901366935
characterization of the opine-utilizing microflora associated with samples of soil and plants.microorganisms utilizing an opine as the sole carbon source were recovered from crown gall tumors, soil, and surface-disinfected potato tubers. the effect of the opines octopine, nopaline, succinamopine, and mannopine as selective substrates was compared with that of the auxin indoleacetic acid. selection on octopine and indoleacetic acid favored the fluorescent pseudomonads, whereas mannopine allowed the frequent recovery of agrobacteria. coryneforms which utilized succinamopine or mannopine we ...199016348265
monoclonal antibodies to ferric pseudobactin, the siderophore of plant growth-promoting pseudomonas putida b10.monoclonal antibodies to ferric pseudobactin, the siderophore (microbial iron transport agent) of plant growth-promoting pseudomonas putida b10, have been developed. three immunoglobulin g subclass 1-type monoclonal antibodies have been characterized. each antibody appears to be unique on the basis of their reactions with ferric pseudobactin and with culture supernatants from other pseudomonads. none of the three cross-reacts with ferric pseudobactin-type siderophores produced by seven other pse ...199016348116
copper resistance gene homologs in pathogenic and saprophytic bacterial species from tomato.copper-resistant strains of xanthomonas campestris pv. vesicatoria, pseudomonas cichorii, pseudomonas putida, pseudomonas fluorescens, and a yellow pseudomonas sp. were isolated from tomato plants or seeds. in southern hybridizations, dna from each strain showed homology with the copper resistance (cop) operon previously cloned from pseudomonas syringae pv. tomato pt23. homology was associated with plasmid and chromosomal dna in x. compestris pv. vesicatoria, p. putida, and the yellow pseudomona ...199016348118
specificity of octopine uptake by rhizobium and pseudomonas strains.the octopine-utilizing strain agrobacterium tumefaciens b6s3 and three nonagrobacteria which had the capacity to utilize this opine were compared for octopine uptake. the characteristics of uptake by rhizobium meliloti a3 and strain b6s3 were similar. in both bacteria, uptake activity was inducible by octopine and by the related opine octopinic acid, and competition assays showed that these two opine substrates were accepted by the same uptake system with an equivalent affinity. cells of pseudom ...199016348194
catalase and superoxide dismutase of root-colonizing saprophytic fluorescent pseudomonads.root-colonizing, saprophytic fluorescent pseudomonads of the pseudomonas putida-p. fluorescens group express similar levels of catalase and superoxide dismutase activities during growth on a sucrose- and amino acid-rich medium. increased specific activities of catalase but not superoxide dismutase were observed during growth of these bacteria on components washed from root surfaces. the specific activities of both enzymes were also regulated during contact of these bacteria with intact bean root ...199016348360
overproduction of glutathione and its derivatives by genetically engineered microbial cells.in order to improve the biotechnological potentials of escherichia coli cells to produce glutathione, s-d-lactoylglutathione and other gamma-glutamyl compounds, the genes for enzymes [gamma-l-glutamyl-l-cysteine synthetase (gsh a) in e. coli b, glutathione synthetase (gsh b) in e. coli b, glyoxalase i (glo i) in pseudomonas putida] were cloned and amplified in e. coli. e. coli b cells transformed with both gsh a and gsh b genes exhibited a high activity in the synthesis of glutathione and other ...199014545903
long-term survival of and plasmid stability inpseudomonas andklebsiella species and appearance of nonculturable cells in agricultural drainage water.one year after introduction into agricultural drainage waterpseudomonas fluorescens r2f (rp4),pseudomonas putida cym318 (prk2501), andklebsiella aerogenes nctc418 (pbr322) could be recovered on agar media. survival of the introduced strains depended on competition with the indigenous microflora, the presence of nutrients, and the availability of air.in contrast tok. aerogenes nctc418 (pbr322), bothpseudomonas species lost their plasmids, as indicated by the consistently lower colony counts on se ...199024196361
catechol 2,3-oxygenase production by genetically engineered escherichia coli and its application to catechol determination.catechol 2,3-oxygenase was produced by escherichia coli, harbouring the recombinant plasmid pbh100 which contained the pheb gene cloned from phenol-degrading pseudomonas putida bh, and was applied for the determination of catechol in the liquor. e. coli jm103 (pbh100) and c600 (pbh100) showed, respectively, about 5 and 8.5 times higher activities than that of p. putida bh. using the crude extract prepared from the culture broth of the recombinant, catechol between 0.1 and 3.0 μg/ml could be dete ...199124425030
pseudomonas putida which can grow in the presence of toluene.a pseudomonas putida strain able to grow in the presence of more than 50% toluene was isolated from soil. the strain was tolerant of other toxic solvents, including aliphatic hydrocarbons, alicyclic hydrocarbons, aromatic hydrocarbons, alcohols, and ethers. the stability of the solvent tolerance of strain ih-2000 was stimulated by addition of mg and ca to the medium containing toluene.199116348494
monohydroxylation of phenol and 2,5-dichlorophenol by toluene dioxygenase in pseudomonas putida f1.[this corrects the article on p. 2650 in vol. 55.].199116348499
enhancement of the potential to utilize octopine in the nonfluorescent pseudomonas sp. strain 92.the nonfluorescent pseudomonas sp. strain 92 requires the presence of a supplementary carbon source for growth on octopine, whereas the spontaneous mutant rb100 has acquired the capacity to utilize this opine as the sole carbon and nitrogen source. insertional mutagenesis of rb100 with transposon tn5 generated mutants which were unable to grow on octopine and others which grew slowly on this substrate. both types of mutants yielded revertants that had regained the ability to utilize octopine. so ...199116348533
rapid immunocapture of pseudomonas putida cells from lake water by using bacterial flagella.monoclonal antibodies to pseudomonas putida paw340 cells were produced. in an enzyme-linked immunosorbent assay (elisa) against whole bacterial cells, a hybridoma cell line termed mlv1 produced a monoclonal antibody that reacted with p. putida paw340 but showed no cross-reaction with 100 medical isolates and 150 aquatic isolates. by elisa, immunogold electron microscopy, and western blot (immunoblot) analysis, mlv1 antibody was found to react with purified bacterial flagella. the surfaces of mag ...199116348416
conditional-suicide containment system for bacteria which mineralize aromatics.a model conditional-suicide system to control genetically engineered microorganisms able to degrade substituted benzoates is reported. the system is based on two elements. one element consists of a fusion between the promoter of the pseudomonas putida tol plasmid-encoded meta-cleavage pathway operon (p(m)) and the laci gene encoding lac repressor plus xyls, coding for the positive regulator of p(m). the other element carries a fusion between the p(tac) promoter and the gef gene, which encodes a ...199116348490
effects of 2,4-dichlorophenol, a metabolite of a genetically engineered bacterium, and 2,4-dichlorophenoxyacetate on some microorganism-mediated ecological processes in soil.a genetically engineered microorganism, pseudomonas putida ppo301(pro103), and the plasmidless parent strain, ppo301, were added at approximately 10 cfu/g of soil amended with 500 ppm of 2,4-dichlorophenoxyacetate (2,4-d) (500 mug/g). the degradation of 2,4-d and the accumulation of a single metabolite, identified by gas chromatography-mass spectrophotometry as 2,4-dichlorophenol (2,4-dcp), occurred only in soil inoculated with ppo301(pro103), wherein 2,4-dcp accumulated to >70 ppm for 5 weeks a ...199116348408
flavonoids released naturally from alfalfa seeds enhance growth rate of rhizobium meliloti.alfalfa (medicago sativa l.) releases different flavonoids from seeds and roots. imbibing seeds discharge 3',4',5,7-substituted flavonoids; roots exude 5-deoxy molecules. many, but not all, of these flavonoids induce nodulation (nod) genes in rhizobium meliloti. the dominant flavonoid released from alfalfa seeds is identified here as quercetin-3-o-galactoside, a molecule that does not induce nod genes. low concentrations (1-10 micromolar) of this compound, as well as luteolin-7-o-glucoside, anot ...199116668056
broad host-range vector for efficient expression of foreign genes in gram-negative bacteria.a broad host-range expression plasmid was constructed comprising the incq replicon, the reca promoter from escherichia coli and the g10-l ribosome binding site (rbs) derived from bacteriophage t7. the structural genes for porcine somatotropin (pst) and e. coli beta-galactosidase (lacz) were used to monitor gene expression in a diverse collection of gram-negative bacterial hosts: escherichia coli, pseudomonas aeruginosa, pseudomonas syringae, pseudomonas putida, pseudomonas fluorescens, pseudomon ...19911367537
molecular cloning of a pseudomonas paucimobilis gene encoding a 17-kilodalton polypeptide that eliminates hcl molecules from gamma-hexachlorocyclohexane.pseudomonas paucimobilis ut26 is capable of growing on gamma-hexachlorocyclohexane (gamma-hch). a genomic library of p. paucimobilis ut26 was constructed in pseudomonas putida by using the broad-host-range cosmid vector pks13. after 2,300 clones were screened by gas chromatography, 3 clones showing gamma-hch degradation were detected. a 5-kb fragment from one of the cosmid clones was subcloned into puc118, and subsequent deletion and gas chromatography-mass spectrometry analyses revealed that a ...19911718942
cloning, sequence and transcriptional analysis of the structural gene for lpd-3, the third lipoamide dehydrogenase of pseudomonas putida.the third lipoamide dehydrogenase structural gene of pseudomonas putida, lpd3, was isolated from a library of p. putida ppg2 dna cloned in escherichia coli tb1. the nucleotide sequence of lpd3 and its flanking regions indicate that lpd3 is not part of an operon, which is unique for a prokaryotic lipoamide dehydrogenase. an open reading frame was found 207 bases upstream from the start of transcription, but is encoded on the strand opposite lpd3. there is no evidence of an open reading frame imme ...19911722146
the ferric-pseudobactin receptor pupa of pseudomonas putida wcs358: homology to tonb-dependent escherichia coli receptors and specificity of the protein.the initial step in the uptake of iron via ferric pseudobactin by the plant-growth-promoting pseudomonas putida strain wcs358 is binding to a specific outer-membrane protein. the nucleotide sequence of the pupa structural gene, which codes for a ferric pseudobactin receptor, was determined. it contains a single open reading frame which potentially encodes a polypeptide of 819 amino acids, including a putative n-terminal signal sequence of 47 amino acids. significant homology, concentrated in fou ...19911646376
microbial metabolism of quinoline and related compounds. viii. xanthine dehydrogenase from a quinoline utilizing pseudomonas putida strain.the xanthine dehydrogenase from pseudomonas putida 86 was purified 68-fold to homogeneity with 47% recovery. sds-polyacrylamide gel electrophoresis of the enzyme revealed two protein bands corresponding to an mr of 87,000 and 52,000. the mr of the native enzyme was calculated to 550,000 by gel chromatography. the enzyme contained 4 atoms of molybdenum, 16 atoms of iron, 16 atoms of acidlabile sulphur and 4 molecules of fad. due to the composition of the cofactors the xanthine dehydrogenase belon ...19911647164
covalent structure of the diheme cytochrome subunit and amino-terminal sequence of the flavoprotein subunit of flavocytochrome c from chromatium vinosum.the complete sequence of the 21-kda cytochrome subunit of the flavocytochrome c (fc) from the purple phototrophic bacterium chromatium vinosum has been determined to be as follows: eptaemltnncagchg thgnsvgpaspsiaqmdpmvfvevmegfksgeias timgriakgystadfekmagyfkqqtyqpakqsf dtaladtgaklhdkycekchveggkpladeedy hilagqwtpylqyamsdfreerrpmekkmaskl rellkaegdagldalfafyasqq. the sequence is the first example of a diheme cytochrome in a flavocytochrome complex. although the locations of the heme binding sites an ...19911649169
expression, localization, and functional analysis of polychlorinated biphenyl degradation genes cbpabcd of pseudomonas putida.genes of pseudomonas putida strains that are capable of degrading polychlorinated biphenyls were cloned in the plasmid vector puc19. the resultant hybrid plasmid, paw6194, contained cbpabcd genes on a 9.0-kb dna fragment that was necessary for the catabolism of polychlorinated biphenyls. these genes were further subcloned on an 8.0-kb hindiii fragment of paw540. degradation of 3-chlorobiphenyl, 2,4-dichlorobiphenyl, and 2,4,5-trichlorobiphenyl into a chloro derivative of benzoic acid was found i ...19911649578
overproduction, purification, and characterization of chlorocatechol dioxygenase, a non-heme iron dioxygenase with broad substrate tolerance.we show here that purified chlorocatechol dioxygenase from pseudomonas putida is able to oxygenate a wide range of substituted catechols with turnover numbers ranging from 2 to 29 s-1. this enzyme efficiently cleaves substituted catechols bearing electron-donating or multiple electron-withdrawing groups in an intradiol manner with kcat/km values between 0.2 x 10(7) and 1.4 x 10(7) m-1 s-1. these unique catalytic properties prompted a comparison with the related but highly specific enzymes catech ...19911649626
functional analysis of the pseudomonas putida regulatory protein catr: transcriptional studies and determination of the catr dna-binding site by hydroxyl-radical footprinting.catr, a lysr family protein, positively regulates the pseudomonas putida catbc operon, which is required for growth on benzoate as a sole carbon source. transcriptional studies show that the catr and catbc promoters are divergent and overlapping by 2 bp. a beta-galactosidase promoter probe vector was constructed to analyze expression from the catr and catbc promoters under induced and uninduced conditions. as predicted, the catbc promoter is expressed only under induced conditions, while the cat ...19911649820
purification of glucose-inducible outer membrane protein oprb of pseudomonas putida and reconstitution of glucose-specific pores.a 43,000 molecular-weight, glucose-inducible, organic acid-repressible protein (oprb) was identified in the outer membrane of pseudomonas putida. oprb was surface expressed in whole cells, had a high beta-sheet content, and was heat modifiable, as demonstrated by 125i-labeling, circular dichroism spectroscopy, and mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. oprb was extracted from outer membrane preparations by using 2% lubrol px with 10 mm edta and purified by deae-se ...19911650338
sequence of the gene (phea) encoding phenol monooxygenase from pseudomonas sp. est1001: expression in escherichia coli and pseudomonas putida.the plasmid pest1412 contains the genes, phea and pheb, encoding phenol monooxygenase (pmo) and catechol 1,2-dioxygenase (c12]), respectively. thse were originally cloned from the plasmid dna of pseudomonas sp. est1001 [kivisaar et al., plasmid 24 (1990) 25-36]. although phea and pheb are cotranscribed using the promoter sequences derived from tn4652 and the level of expression of c120 activities from pest1412 was equal both in escherichia coli and in pseudomonas putida, the level of pmo activit ...19911650730
microbial metabolism of quinoline and related compounds. x. the molybdopterin cofactors of quinoline oxidoreductases from pseudomonas putida 86 and rhodococcus spec. b1 and of xanthine dehydrogenase from pseudomonas putida 86.the bis(carboxamidomethyl) derivatives of the molybdenum cofactors in three eubacterial molybdo-iron/sulphur-flavoproteins were examined. the quinoline oxidoreductases from pseudomonas putida 86 and rhodococcus spec. b1 contain molybdopterin cytosine dinucleotide. in xanthine dehydrogenase from pseudomonas putida 86, however, only molybdopterin was found. the bis(carboxamidomethyl) derivatives of all three enzymes were treated with nucleotide pyrophosphatase, but only those of the quinoline oxid ...19911657036
genetic organization and regulation of the xylose degradation genes in streptomyces rubiginosus.the xylose isomerase (xyla) and the xylulose kinase (xylb) genes from streptomyces rubiginosus were isolated, and their nucleotide sequences were determined. the xyla and xylb genes encode proteins of 388 and 481 amino acids, respectively. these two genes are transcribed divergently from within a 114-nucleotide sequence separating the coding regions. regulation of the xyl genes in s. rubiginosus was examined by fusing their promoters to the pseudomonas putida catechol dioxygenase gene and integr ...19911657868
identification of a novel composite transposable element, tn5280, carrying chlorobenzene dioxygenase genes of pseudomonas sp. strain p51.analysis of one of the regions of catabolic plasmid pp51 which encode chlorobenzene metabolism of pseudomonas sp. strain p51 revealed that the tcba and tcbb genes for chlorobenzene dioxygenase and dehydrogenase are located on a transposable element, tn5280. tn5280 showed the features of a composite bacterial transposon with iso-insertion elements (is1066 and is1067) at each end of the transposon oriented in an inverted position. when a 12-kb hindiii fragment of pp51 containing tn5280 was cloned ...19911657878
conjugational transfer of recombinant dna in cultures and in soils: host range of pseudomonas putida tol plasmids.recombinant tol plasmid pwwo-eb62 allows pseudomonas putida to grow on p-ethylbenzoate. this plasmid can be transferred to other microorganisms, and its catabolic functions for the metabolism of alkylbenzoates are expressed in a limited number of gram-negative bacteria, including members of pseudomonad rrna group i and escherichia coli. transfer of the recombinant plasmid to erwinia chrysanthemi was observed, but transconjugants failed to grow on alkylbenzoates because they lost catabolic functi ...19911660698
nucleotide sequence of xyle from the tol pdk1 plasmid and structural comparison with isofunctional catechol-2,3-dioxygenase genes from tol, pww0 and nah7.detailed restriction and nucleotide sequence analysis of the pseudomonas putida tol plasmid pdk1 xyle gene revealed significant homology with isofunctional xyle (81.5%) and nahh (78.0%) genes from the tol pww0 and nah7 plasmids. the highest degrees of nucleotide and apparent amino acid conservation (82.2 and 86.4%, respectively) among all three genes were found to exist within a region comprising 264 nucleotides encoding the c terminus. a comparison of localized regions revealed significantly gr ...19911672868
cloning and sequencing the urocanase gene (hutu) from pseudomonas putida.a clone harbouring the entire urocanase gene (hutu) was obtained from a genomic library of pseudomonas putida using oligonucleotide probes synthesised on the basis of known flanking sequences. one subunit of urocanase consists of 556 amino acids and has a molecular mass of 60,771 da.19911677899
monitoring a genetically engineered bacterium in a freshwater environment by rapid enzymatic amplification of a synthetic dna "number-plate".in order to set up a sensitive and reliable detection method to monitor environmentally released genetically engineered microorganisms (gems) a 72-bp, double-stranded dna fragment has been built by annealing and ligating four synthetic oligonucleotides. binding sites for two 20-mer oligonucleotides are situated inside the dna fragment, flanking the centre. into the central part of the construction a 30-nucleotide identification sequence has been fitted. thanks to the presence of the two oligonuc ...19911369367
cloning and characterization of plasmid-encoded genes for the degradation of 1,2-dichloro-, 1,4-dichloro-, and 1,2,4-trichlorobenzene of pseudomonas sp. strain p51.pseudomonas sp. strain p51 is able to use 1,2-dichlorobenzene, 1,4-dichlorobenzene, and 1,2,4-trichlorobenzene as sole carbon and energy sources. two gene clusters involved in the degradation of these compounds were identified on a catabolic plasmid, pp51, with a size of 110 kb by using hybridization. they were further characterized by cloning in escherichia coli, pseudomonas putida kt2442, and alcaligenes eutrophus jmp222. expression studies in these organisms showed that the upper-pathway gene ...19911987135
molecular characterization and expression analysis of the anthranilate synthase gene of pseudomonas syringae subsp. savastanoi.the trpe gene, which encodes the large component of the enzyme anthranilate synthase, was isolated from a pseudomonas syringae subsp. savastanoi (p. savastanoi) cosmid library. cosmids that complemented an escherichia coli trpe mutation contained a gene whose product is 86% homologous at the deduced amino acid level to trpe of pseudomonas aeruginosa and pseudomonas putida. amino acid sequence comparison with other trpe sequences revealed the existence of conserved regions between the procaryotic ...19911987141
comparison of benzyl alcohol dehydrogenases and benzaldehyde dehydrogenases from the benzyl alcohol and mandelate pathways in acinetobacter calcoaceticus and from the tol-plasmid-encoded toluene pathway in pseudomonas putida. n-terminal amino acid sequences, amino acid compositions and immunological cross-reactions.1. n-terminal sequences were determined for benzyl alcohol dehydrogenase, benzaldehyde dehydrogenase i and benzaldehyde dehydrogenase ii from acinetobacter calcoaceticus n.c.i.b. 8250, benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase encoded by the tol plasmid pww53 in pseudomonas putida mt53 and yeast k(+)-activated aldehyde dehydrogenase. comprehensive details of the sequence determinations have been deposited as supplementary publication sup 50161 (5 pages) at the british library d ...19911989592
metabolism of poly(3-hydroxyalkanoates) (phas) by pseudomonas oleovorans. identification and sequences of genes and function of the encoded proteins in the synthesis and degradation of pha.pseudomonas oleovorans accumulates poly(3-hydroxyalkanoates) (phas) after growth on medium chain length hydrocarbons. large amounts of this polyester are synthesized when cells are grown under nitrogen-limiting conditions. when nitrogen is resupplied in the medium, the accumulated pha is degraded. in this paper, we describe mutants which are defective in the synthesis or in the degradation of pha. these mutants were used to select dna fragments which encode pha polymerases and a pha depolymerase ...19911989978
bradyrhizobium japonicum has two differentially regulated, functional homologs of the sigma 54 gene (rpon).recognition of -24/-12-type promoters by rna polymerase requires a special sigma factor, sigma 54 (rpon ntra glnf). in the nitrogen-fixing soybean symbiont bradyrhizobium japonicum, two functional, highly conserved rpon genes (rpon1 and rpon2) were identified and sequenced. the two predicted b. japonicum rpon protein sequences were 87% identical, and both showed different levels of homology to the rpon proteins of other bacteria. downstream of rpon2 (but not of rpon1), two additional open readin ...19911991712
nucleotide sequence analysis of the pseudomonas putida ppg7 salicylate hydroxylase gene (nahg) and its 3'-flanking region.gene nahg of naphthalene/salicylate catabolic plasmid nah7 encodes a protein of molecular weight 45,000, salicylate hydroxylase. this enzyme catalyzes the formation of catechol from salicylate, a key intermediate in naphthalene catabolism. dna sequence analysis of the 3.1-kilobase hindiii fragment containing the nahg locus reveals an open reading frame (orf) of 1305 base pairs that corresponds to a protein of 434 amino acid residues. the predicted amino acid sequence of salicylate hydroxylase is ...19911993181
freeze-substitution of gram-negative eubacteria: general cell morphology and envelope profiles.freeze-substitution was performed on strains of escherichia coli, pasteurella multocida, campylobacter fetus, vibrio cholerae, pseudomonas aeruginosa, pseudomonas putida, aeromonas salmonicida, proteus mirabilis, haemophilus pleuropneumoniae, caulobacter crescentus, and leptothrix discophora with a substitution medium composed of 2% osmium tetroxide and 2% uranyl acetate in anhydrous acetone. a thick periplasmic gel ranging from 10.6 to 14.3 nm in width was displayed in e. coli k-12, k30, and hi ...19911999383
primary structure of xylene monooxygenase: similarities to and differences from the alkane hydroxylation system.xylene monooxygenase, encoded by the tol plasmid of pseudomonas putida, catalyzes the oxidation of toluene and xylenes and consists of two different subunits encoded by xyla and xylm. in this study, the complete nucleotide sequences of these genes were determined and the amino acid sequences of the xyla and xylm products were deduced. the xylm sequence had a 25% homology with alkane hydroxylase, which catalyzes the omega-hydroxylation of fatty acids and the terminal hydroxylation of alkanes. the ...19911999388
construction of tracer plasmids for bacillus sphaericus 1593 utilizing the xyle gene from pseudomonas putida.genetically engineered microorganisms (gems) released into the environment must be traceable in order to accurately assess their impact on the area of release. tracer genes other than those that introduce antibiotic resistance are preferred for use in identifying genetically engineered strains. in this study, we describe the construction of a series of tracer plasmids for use in bacillus sphaericus using the xyle gene from the pseudomonas putida tol plasmid. this gene codes for the enzyme catech ...19912002245
fluorometric determination of phenylacetyl-coa ligase from pseudomonas putida: a very sensitive assay for a newly described enzyme.phenylacetyl-coa ligase (amp-forming) from pseudomonas putida is a newly described enzyme (martinez-blanco, h., reglero, a., rodriguez-aparicio, l.b. and luengo, j.m. (1990) j. biol. chem. 265, 7084-7090) specifically involved in the catabolism of phenylacetic acid. this enzyme catalyzes the formation of phenylacetyl-coa in the presence of atp, coa, mg2+ and phenylacetic acid. a rapid method of assaying this enzyme in partially purified preparations has been developed by coupling this reaction w ...19912009287
microbial degradation of the morphine alkaloids. purification and characterization of morphine dehydrogenase from pseudomonas putida m10.the nadp(+)-dependent morphine dehydrogenase that catalyses the oxidation of morphine to morphinone was detected in glucose-grown cells of pseudomonas putida m10. a rapid and reliable purification procedure involving two consecutive affinity chromatography steps on immobilized dyes was developed for purifying the enzyme 1216-fold to electrophoretic homogeneity from p. putida m10. morphine dehydrogenase was found to be a monomer of mr 32,000 and highly specific with regard to substrates, oxidizin ...19912012614
sequence of the plasmid-encoded catechol 1,2-dioxygenase-expressing gene, pheb, of phenol-degrading pseudomonas sp. strain est1001.phenol monooxygenase (pmo) and catechol 1,2-dioxygenase (c12o), the two first enzymes of the phenol-degradation pathways, are encoded by a 3.4-kb dna fragment cloned from pseudomonas sp. est1001 plasmid dna. we have previously shown that activation of the cloned genes in pseudomonas putida paw85 is controlled by insertion of the 17-kb transposon, tn4652, from the host chromosome into the plasmid carrying these genes [kivisaar et al. plasmid 24 (1990) 25-36]. transcription of the dna encoding pmo ...19912013408
sequence analysis of the pseudomonas sp. strain p51 tcb gene cluster, which encodes metabolism of chlorinated catechols: evidence for specialization of catechol 1,2-dioxygenases for chlorinated substrates.pseudomonas sp. strain p51 contains two gene clusters located on catabolic plasmid pp51 that encode the degradation of chlorinated benzenes. the nucleotide sequence of a 5,499-bp region containing the chlorocatechol-oxidative gene cluster tcbcdef was determined. the sequence contained five large open reading frames, which were all colinear. the functionality of these open reading frames was studied with various escherichia coli expression systems and by analysis of enzyme activities. the first g ...19912013566
survival and catabolic activity of natural and genetically engineered bacteria in a laboratory-scale activated-sludge unit.the survival of selected naturally occurring and genetically engineered bacteria in a fully functional laboratory-scale activated-sludge unit (asu) was investigated. the effect of the presence of 3-chlorobenzoate (3cb) on the survival of pseudomonas putida uwc1, with or without a chimeric plasmid, pd10, which encodes 3cb catabolism, was determined. p. putida uwc1(pd10) did not enhance 3cb breakdown in the asu, even following inoculation at a high concentration (3 x 10(8) cfu/ml). the emergence o ...19912014987
an upstream xylr- and ihf-induced nucleoprotein complex regulates the sigma 54-dependent pu promoter of tol plasmid.transcription from promoter pu of the upper catabolic operon of the pseudomonas putida tol plasmid which specifies conversion of toluene/xylenes to benzoate/toluates is activated by the tol-encoded regulator xylr protein in the presence of substrates of the catabolic pathway and in conjunction with the sigma 54(ntra)-containing form of rna polymerase. this regulatory circuit was faithfully reproduced in escherichia coli in single copy gene dosage by integrating the corresponding controlling dete ...19912022186
survival in soils of an herbicide-resistant pseudomonas putida strain bearing a recombinant tol plasmid.pseudomonas putida eez15(pww0-eb62) is a phosphinothricin (ppt)-resistant strain with a recombinant tol plasmid which allows the strain to grow on p-ethylbenzoate. the survival of this strain in sterile agricultural soils depends on the physicochemical properties of the soil. the recombinant pww0-eb62 plasmid and its catabolic functions were stable for periods of up to 1 month in bacteria introduced in unamended soils and only conferred selective advantage to the host bacteria without the plasmi ...19912036014
[phage-stable mutants of pseudomonas putida: new types of stability].more than 170 phage-resistant mutants (prm) of the first order of pseudomonas putida strain ppg1 were obtained using newly isolated and previously described bacteriophages specific for this strain. according to the results of analysis of resistance of the mutants to each of 31 phages of ppg1 strain and 8 phages of the ppn strain, the prm strains were distributed into 20 groups. in most cases, the reason for resistance is loss of absorption capacity of bacteria. however, no direct relation betwee ...19912037253
observation of the o-o stretching raman band for cytochrome p-450cam under catalytic conditions.dioxygen stretching (voo) raman band was observed for the oxy form of pseudomonas putida cytochrome p-450 (p-450cam) generated at room temperature under catalytic conditions, that is, in the presence of d-camphor, beta-nadh, putidaredoxin, and putidaredoxin reductase, by using the mixed flow transient raman apparatus. at the same time the visible absorption spectra were monitored for the transient species. it was found that the voo frequency is little altered by binding of putidaredoxin to p-450 ...19912037577
primary and secondary structural patterns in eukaryotic cytochrome p-450 families correspond to structures of the helix-rich domain of pseudomonas putida cytochrome p-450cam. indications for a similar overall topology.an extensive sequence analysis of the eukaryotic cytochrome p-450 (p-450) protein families was conducted with a view to identifying conserved regions that might be related to secondary structural features in the pseudomonas putida camphor hydroxylase (p-450cam). all sequences available on-line were collected, classified and aligned within families. distinctively different sequences were chosen from each of seven eukaryotic families, and an unbiased multi-alignment was constructed. profile patter ...19912040297
characterization of the pseudomonas sp. strain p51 gene tcbr, a lysr-type transcriptional activator of the tcbcdef chlorocatechol oxidative operon, and analysis of the regulatory region.plasmid pp51 of pseudomonas sp. strain p51 contains two gene clusters encoding the degradation of chlorinated benzenes, tcbab and tcbcdef. a regulatory gene, tcbr, was located upstream and divergently transcribed from the chlorocatechol oxidative gene cluster tcbcdef. the tcbr gene was characterized by dna sequencing and expression studies with escherichia coli and pet8c and appeared to encode a 32-kda protein. the activity of the tcbr gene product was analyzed in pseudomonas putida kt2442, in w ...19912050630
structure-function relationships of human aromatase cytochrome p-450 using molecular modeling and site-directed mutagenesis.the conversion of androgens to estrogens is catalyzed by an enzyme complex named aromatase, which consists of a form of cytochrome p-450, aromatase cytochrome p-450 (cytochrome p-450arom), and the flavoprotein, nadph-cytochrome p-450 reductase. as a first step toward investigation of the structure-function relationships of cytochrome p-450arom, we have used computer modeling to align the amino acid sequence of cytochrome p-450arom with that of cytochrome p-450cam from pseudomonas putida and thus ...19912050688
molybdenum-dependent degradation of quinoline by pseudomonas putida chin ik and other aerobic bacteria.eighteen different aerobic bacteria were isolated which utilized quinoline as sole source of carbon, nitrogen, and energy. attempts were unsuccessful at isolating anaerobic quinoline-degrading bacteria. the optimal concentration of quinoline for growth was in the range of 2.5 to 5 mm. some organisms excreted 2-hydroxyquinoline as the first intermediate. hydroxylation of quinoline was catalyzed by a dehydrogenase which was induced in the presence of quinoline or 2-hydroxyquinoline. quinoline dehy ...19912059099
a new family of bacterial regulatory proteins.a new family of bacterial regulatory proteins has been identified by sequence similarity. the family contains the repressor of the bacillus subtilis gluconate operon (gntr), the regulators for histidine utilization in pseudomonas putida (hutcpp) and klebsiella aerogenes (hutcka), the repressor (fadr) of fatty acid degradation in escherichia coli, a regulator involved in the conjugal transfer of the broad host range plasmid pij101 (kora), and three proteins of unidentified function in e. coli (ge ...19912060763
divergent evolution of chloroplast-type ferredoxins.the tol plasmid pww0 of pseudomonas putida encodes a set of enzymes required for the oxidation of toluene to krebs cycle intermediates. the structural genes for these enzymes are encoded in two operons which comprise the xylcmabn and xylxyzltegfjqkih genes, respectively. the function of the xylt gene has not yet been identified. the nucleotide sequence of xylt was determined in this study and putative gene product was shown to contain a sequence characteristic for chloroplast-type ferredoxins. t ...19912065785
[plasmid r68.45 and s-a transduction by the temperate bacteriophage 59 of erwinia carotovora 268].temperature bacteriophage 59 of erwinia carotovera 268 had transduced extrachromosomal dna: plasmids of r68.45 and s-a. before plasmid transduction experiments the suitable donor strains of indicator culture erwinia horticola 450 harbouring r68.45 and s-a were created. the frequency of plasmid r68.45 transfer from pseudomonas putida to e. horticola 450-8 by conjugation was equal to 5 x 10(-8) per a donor cell and in the case of s-a--from e. coli c600 for the same recipient cells--was 2 x 10(-6). ...19912067419
[the effect of amino acids in the asparagine family on the aspartate kinase and homoserine dehydrogenase of ethionine-resistant mutants of pseudomonas putida].the activity of aspartate kinase and homoserin dehydrogenase from ethionine resistant mutants pseudomonas putida 25 and 6 have been studied as affected by amino acids from the family of asparagine. they are characterized by a capacity to the surplus synthesis of methionine. it is shown that mutants have negative regulation of the level of activity of the studied enzymes. it is supposed that the mutations (or mutation) could take place which affected properties of enzymes, which participated dire ...19912067422
[characteristics of the r-plasmid pm3 (incp-9) of a broad circle of hosts].a new broad host range plasmid pm3 (incp-9) was found in a facultative methylotrophic bacteria pseudomonas putida and described. the pm3 plasmid is characterized by thermo-instability in enterobacteriaceae family of bacteria at 36 degrees c or higher temperatures. it is also unable to be inherited as an autonomous element in the obligate methylotrophic bacteria methylobacillus m75. the peculiarities of plasmid inheritance make possible to use it as a tool for genetic research, for instance, to c ...19911745275
subcloning of bph genes from pseudomonas testosteroni b-356 in pseudomonas putida and escherichia coli: evidence for dehalogenation during initial attack on chlorobiphenyls.the bpha, -b, -c, and -d genes from pseudomonas testosteroni b-356 were mapped to a 5.5-kb dna fragment of cloned plasmids pda1 and pda2 by use of deletion and insertion mutants of these plasmids. the expression of each of these genes was evaluated in escherichia coli and in pseudomonas putida, and it was found that the bphc and bphd genes are well expressed in both e. coli and p. putida cells while the bpha and bphb genes are very poorly expressed in e. coli, even when placed downstream of a ta ...19911746948
septicaemia and septic arthritis due to pseudomonas putida in a neutropenic patient. 19911753153
"in vitro" synthesis of different naturally-occurring, semisynthetic and synthetic penicillins using a new and effective enzymatic coupled system.forty-seven different penicillins, including some of great clinical importance, have been synthesized "in vitro" by coupling the newly described enzyme phenylacetyl-coa ligase (pcl) from pseudomonas putida and acyl-coa: 6-aminopenicillanic acid (6-apa) acyltransferase (at) from penicillium chrysogenum. incubations were carried out at 30 degrees c in 50 mm hcl-tris buffer ph 8.0. the reaction mixtures contained 6-apa, coa, atp, dithiothreitol, mg2+ and the corresponding penicillin side-chain prec ...19911761422
synthesis of 3-furylmethylpenicillin using an enzymatic procedure.3-furylmethylpenicillin was synthesized in vitro from 3-furylacetic acid, 6-aminopenicillanic acid (6-apa), coa, atp and mg2+. the reaction was catalyzed in two steps by the enzymes phenyl-acetyl-coa ligase (pcl) from pseudomonas putida and acyl-coa: 6-apa acyltransferase (at) from penicillium chrysogenum. pcl catalyzes the activation of 3-furylacetic acid to 3-furylacetyl-coa (3-f-coa) and at acylates the amino group of 6-apa with the 3-furylacetyl moiety of 3-f-coa, releasing coa and 3-furylme ...19911778415
[cloning of genes degrading 3-chlorobenzoate from pseudomonas putida strain 87].the ability of pseudomonas putida strain 87 to catabolize 3-chlorobenzoate was shown to be mediated by genes of pbs109 plasmid. the plasmid may be transferred by conjugation into p. aeruginosa pao2175. it seems possible that the pbs109 plasmid codes for pyrocatechase ii specific for halogenated catechol, but not catechol. the genes specifying utilization of 3-chlorobenzoate from pbs109 plasmid were cloned in the 5.5 kb bgiii fragment by using broad-host cloning system. the resulting pbs110 plasm ...19911778448
biotransformation of nitrobenzene by bacteria containing toluene degradative pathways.nonpolar nitroaromatic compounds have been considered resistant to attack by oxygenases because of the electron withdrawing properties of the nitro group. we have investigated the ability of seven bacterial strains containing toluene degradative pathways to oxidize nitrobenzene. cultures were induced with toluene vapor prior to incubation with nitrobenzene, and products were identified by high-performance liquid chromatography and gas chromatography-mass spectrometry. pseudomonas cepacia g4 and ...19911781679
metabolism of and inhibition by chlorobenzoates in pseudomonas putida p111.pseudomonas putida p111 was isolated by enrichment culture on 2,5-dichlorobenzoate and was also able to grow on 2-chloro-, 3-chloro-, 4-chloro-, 2,3-dichloro-, 2,4-dichloro-, and 2,3,5-trichlorobenzoates. however, 3,5-dichlorobenzoate completely inhibited growth of p111 on all ortho-substituted benzoates that were tested. when 3,5-dichlorobenzoate was added as a cosubstrate with either 3- or 4-chlorobenzoate, cell yields and chloride release were greater than those observed from growth on either ...19911781694
anaerobic growth and cyanide synthesis of pseudomonas aeruginosa depend on anr, a regulatory gene homologous with fnr of escherichia coli.anaerobic growth of pseudomonas aeruginosa on nitrate or arginine requires the anr gene, which codes for a positive control element (anr) capable of functionally complementing an fnr mutation in escherichia coli. the anr gene was sequenced; it showed 51% identity with the fnr gene at the amino acid sequence level. four cysteine residues known to be essential in the fnr protein are conserved in anr. the anr gene product (deduced mr 27,129) was visualized by the maxicell method and migrated like a ...19911787798
microbial metabolism of quinoline and related compounds. xii. isolation and characterization of the quinoline oxidoreductase from rhodococcus spec. b1 compared with the quinoline oxidoreductase from pseudomonas putida 86.quinoline oxidoreductase from rhodococcus spec. b1 was purified 39-fold to apparent homogeneity in a 5-step procedure with a recovery of 26%. the mr of the native enzyme as determined by gel chromatography was 300,000. sds polyacrylamide gel electrophoresis of the enzyme revealed 3 protein bands corresponding to mr 82,000, 32,000, and 18,000. the enzyme contains 1.3 atoms of molybdenum, 8 atoms of iron, 8 atoms of acid-labile sulphur, 2 molecules of fad and 2 molecules of molybdopterin cytosine ...19911789933
phenotypic variation of pseudomonas putida and p. tolaasii affects the chemotactic response to agaricus bisporus mycelial exudate.the chemotactic response of wild-type pseudomonas putida and p. tolaasii, and a phenotypic variant of each species, to agaricus bisporus mycelial exudate was examined. both p. putida, the bacterium responsible for initiating basidiome development of a. bisporus, and p. tolaasii, the causal organism of bacterial blotch disease of the mushroom, displayed a positive chemotactic response to casamino acids and to a. bisporus mycelial exudate. the response was both dose- and time-dependent and marked ...19911791431
phenotypic variation of pseudomonas putida and p. tolaasii affects attachment to agaricus bisporus mycelium.the effect of phenotypic variation on attachment of pseudomonas tolaasii and p. putida to agaricus bisporus mycelium was investigated. quantitative studies demonstrated the ability of each isolate to attach rapidly and firmly to a. bisporus mycelium and significant differences in attachment of wild-type and phenotypic variant strains were observed. this was most pronounced in p. tolaasii, where the percentage attachment of the wild-type form was always greater than that of the phenotypic variant ...19911791432
isolation and characterization of pseudomonas putida mutants affected in arginine, ornithine and citrulline catabolism: function of the arginine oxidase and arginine succinyltransferase pathways.pseudomonas putida mutants impaired in the utilization of arginine are affected in either the arginine succinyltransferase pathway, the arginine oxidase route, or both. however, mutants affected in one of the pathways still grow on arginine as sole carbon source. analysis of the products excreted by both wild-type and mutant strains suggests that arginine is mainly channelled by the oxidase route. proline non-utilizing mutants are also affected in ornithine utilization, confirming the role of pr ...19911791443
dna sequence determination of the tol plasmid (pwwo) xylgfj genes of pseudomonas putida: implications for the evolution of aromatic catabolism.the meta operon of the pseudomonas putida tol plasmid (pwwo) encodes all enzymes of a meta-cleavage pathway for the metabolism of benzoic acids to krebs-cycle intermediates. we have determined and analysed the nucleic acid sequence of a 3442 bp region of the meta operon containing the xyl-gfj genes whose products are involved in the post meta-ring fission transformation of catechols. homology analysis of the xylgfj gene products revealed evidence of biochemical relatedness, suggested enzymatic m ...19911791759
[the autoselection of neustonic forms of bacteria].self-breeding of neuston forms of methylobacterium sp., pseudomonas putida bc-2, alcaligenes paradoxus bc-1, bacillus thuringiensis var. israilensis bacteria as well as of a mixed culture of methylotrophs is shown possible. in spite of ability of hydrophobicity of the cell surface the suggested method of self-breeding may be used to perfect properties of larvicidal biopreparations, and bacterial preparations which intensify self-purification of water bodies.19911791780
[quantitation of acinetobacter calcoaceticus in mixed bacterial cultures by an enzyme immunoassay].an enzyme-linked immunosorbent assay using polyclonal antibodies from rabbits has been developed for quantification of acinetobacter calcoaceticus. bacteria were added to the wells of a microtiter plate coated with anti-acinetobacter immunoglobulin. for detecting bound cells the peroxidase-labelled immunoglobulin fraction was used. over a distinct range there is a linear correlation between bound bacteria and measured absorbance allowing a quantification of bacteria in an order from 10(7) to 10( ...19911793089
highly conserved coding sequences in polychlorinated biphenyl (pcb)-degraders of pseudomonas pseudoalcaligenes kf707 and pseudomonas putida kf715.sixteen strains that utilize biphenyl or polychlorinated biphenyl (pcb) as the sole source of carbon and energy have previously been isolated. nucleotide sequence of the bphabcxd operon (11 kb) of pseudomonas pseudoalcaligenes kf707 has now been determined. when bphd region is compared with the previously reported bphd sequence of another pcb-degrader, pseudomonas putida kf715, homologies at the level of nucleotides as well as amino acids are as high as 96%. moreover, both bphabcxd operon (kf707 ...19911842046
three-dimensional structure of p-cresol methylhydroxylase (flavocytochrome c) from pseudomonas putida at 3.0-a resolution.p-cresol methylhydroxylase (pcmh) isolated from pseudomonas putida is an alpha 2 beta 2 tetramer of approximate subunit mr 49,000 and 9,000. it is a flavocytochrome c containing covalently bound fad in the larger subunit and covalently bound heme in the smaller. crystals in space group p2(1)2(1)2(1) with unit-cell parameters a = 140.3 a, b = 130.6 a, and c = 74.1 a contain one full molecule per asymmetric unit and diffract anisotropically to about 2.8-a resolution in two directions and to about ...19911846290
a series of wide-host-range low-copy-number vectors that allow direct screening for recombinants.a series of controlled expression vectors was constructed based on the wide-host-range plasmid pmmb66eh. some of these new vectors code for the alpha-peptide of beta-galactosidase and allow the direct screening of recombinant clones by inactivation of alpha-complementation. the bla gene was replaced in some plasmids by the cat gene of tn9 coding for chloramphenicol resistance, extending the use into beta-lactam-resistant strains. they all feature either the tac or taclac (tac-lac uv5 in tandem) ...19911847347
[localization of camphor degradative plasmids on the chromosome of pseudomonas putida strains paw].camphor degradative plasmids (cam, prk1) are preferentially situated on chromosomes of pseudomonas putida strains paw. after having been transferred into cam+ strains, the tol plasmid pwwo dissociates into the cryptic plasmid pwwo-8 and chromosome-borne transposon tn4651. the opposite situation, i.e. reconstruction of the tol plasmid pwwo from the cryptic plasmid pwwo-8 and chromosome-borne catabolic operons of the pwwo plasmid has been described. cam- derivatives of the cam plasmid were obtaine ...19911855659
a predicted three-dimensional structure of human cytochrome p450: implications for substrate specificity.a three-dimensional structure for human cytochrome p450ia1 was predicted based on the crystal coordinates of cytochrome p450cam from pseudomonas putida. as there was only 15% residue identity between the two enzymes, additional information was used to establish an accurate sequence alignment that is a prerequisite for model building. twelve representative eukaryotic sequences were aligned and a net prediction of secondary structure was matched against the known alpha-helices and beta-sheets of p ...19911857713
binding sites of pyridine in cytochrome p-450cam.careful titration of oxidized cytochrome p-450cam from pseudomonas putida with pyridine revealed deviations of the eadie plot from linearity in the substrate-bound as well as in the substrate-free protein. a binding model which assumes two binding sites for pyridine--the iron and the camphor binding site--is able to describe completely the nonlinear eadie plot.19911859821
hydroxylation of o-halogenophenol and o-nitrophenol by salicylate hydroxylase.salicylate hydroxylase [ec 1.14.13.1] from pseudomonas putida catalyzed the formation of catechol from substrate analogues such as o-nitro-, o-amino-, o-iodo-, o-bromo-, and o-chloro-phenol by removing the ortho-substituted groups. they are converted into nitrite, ammonia, and halide ions, respectively. kinetic parameters of these reactions were determined by spectrophotometric and polarographic methods. hydroxylation of o-nitro- or o-iodophenol proceeds with the unusual stoichiometry of 2:1:1 f ...19911864847
degradation of l-djenkolate catalyzed by s-alkylcysteine alpha,beta-lyase from pseudomonas putida.s-alkylcysteine alpha, beta-lyase [ec 4.4.1.6] of pseudomonas putida catalyzes alpha,beta-elimination of l-djenkolate [3,3'-methylenedithiobis(2-aminopropionic acid)] to produce pyruvate, ammonia, and s-(mercaptomethyl)cysteine initially. secondly, s-(mercaptomethyl)-cysteine, which was identified in the form of s-(mercaptomethyl)cysteine thiolactone and s-(2-thia-3-carboxypropyl)cysteine in the absence and presence of iodoacetic acid, respectively, is decomposed enzymatically to pyruvate, ammon ...19911869519
assay for detection and enumeration of genetically engineered microorganisms which is based on the activity of a deregulated 2,4-dichlorophenoxyacetate monooxygenase.an assay system was developed for the enumeration of genetically engineered microorganisms expressing a deregulated 2,4-dichlorophenoxyacetate (tfd) monooxygenase, which converts phenoxyacetate (paa) to phenol. in paa-amended cultures of pseudomonas aeruginosa pao1c(pro103) and pseudomonas putida ppo301(pro103), strains which express a deregulated tfd monooxygenase, phenol production was proportional to cell number. phenol was reacted, under specific conditions, with a 4-aminoantipyrine dye to f ...19911872608
cloning and characterization of an extracellular temperature-labile serine protease gene from aeromonas hydrophila.aeromonas virulence is thought to depend on multigenic functions. the gene for an extracellular protease from aeromonas hydrophila so2/2 was cloned in escherichia coli c600-1 by using pij860, bifunctional plasmid, as a vector. the gene encodes for a temperature-labile serine protease (p2) with a molecular mass of approx. 68 kda which is highly inhibited by pmsf. the gene was expressed in streptomyces lividans 1326 by transforming protoplasts with the original clone ppa2. we were also able to tra ...19911874394
activation of the pseudomonas tol plasmid upper pathway operon. identification of binding sites for the positive regulator xylr and for integration host factor protein.expression of the pseudomonas putida tol plasmid upper pathway operon requires a promoter that belongs to the -12/-24 class. stimulation of transcription from this promoter is positively controlled by the effector-activated xylr protein and requires a form of rna-polymerase holoenzyme containing the rpon-encoded sigma factor, sigma 54. xylr-dependent stimulation of transcription from the pseudomonas tol upper pathway promoter was examined using deletions, insertions, and in vivo dimethyl sulfate ...19911874736
[cloning of the arthrobacter globiformis fcba gene for dehalogenase and construction of a hybrid pathway of 4-chlorobenzoic acid degradation in pseudomonas putida].the artrobacter globiformis kzt1 fcba gene responsible for dehalogenase (4-chlorobenzoate-4-hydroxylase) activity was cloned in escherichia coli and pseudomonas putida cells. the character of the fcba gene expression was studied. notwithstanding amplification of the gene dose and control of the inducible plac promoter, the level of substrate dehalogenation by recombinant e. coli strains was lower, as compared with that in the original kzt1 strain. cloning of the fcba gene in p. putida kz6r cells ...19911879677
[genetic control of tryptophan hypersynthesis in regulatory mutants of pseudomonas putida].the 6-fluorotryptophan resistant mr1 mutant was obtained from pseudomonas putida m30 (tyr- phe-) strain. the mutant was able to excrete tryptophan (60 micrograms/ml) and has derepressed arof gene encoding 3-deoxy-d-arabinoheptulosonate-7-phosphate synthase. the mutation isolated was identified as aror with the help of cloning early arof gene of p. putida. on the next step of selection, regulatory mutant mr2 was obtained producing 240 micrograms/ml of tryptophan. the mr2 has derepressed unlinked ...19911879681
[clinical evaluation of cefpirome in children].cefpirome (cpr, hr 810) was clinically evaluated for its efficacy and safety in 11 patients with ages from 4 months to 11 years with bacterial infection. the results obtained are summarized as follows. 1. cpr was administered to 6 patients with bronchopneumonia, a patient with pneumonia, a patient with tonsillitis, 2 patients with acute pharyngitis and a patient with suppurative parotitis at daily dosage levels ranging 55.5-91.7 mg/kg, divided into 3 using intravenous bolus injection or 30 minut ...19911880923
construction of cloning cartridges for development of expression vectors in gram-negative bacteria.a cloning cartridge was constructed that can be inserted into a plasmid of choice to form an expression vector in which gene expression is inducible with an inexpensive inducer, sodium salicylate, at low concentrations. this cartridge consists of a 3.6-kb restriction fragment which contains the positive regulatory gene nahr from plasmid nah7, a promoter, pg, that nahr regulates, a multiple cloning site, a transcription terminator, and a gene conferring tetracycline resistance. within promoter pg ...19911885513
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