metabolism of basic amino acids in pseudomonas putida. properties of the inducible lysine transport system. 19715547703
photoactivation of urocanase in pseudomonas putida: factors influencing activation. 19715549144
metabolism of basic amino acids in pseudomonas putida. catabolism of lysine by cyclic and acyclic intermediates. 19715554291
metabolism of basic amino acids in pseudomonas putida. intermediates in l-arginine catabolism. 19715570437
the breakdown of tropic acid in pseudomonas putida strain l. i. utilization of various substrates; the conversion of tropic acid into phenylacetic acid. 19715573355
new regulatory mutation affecting some of the tryptophan genes in pseudomonas putida.three indole analogues, 5-methylindole, 5-fluoroindole, and 7-methylindole, and the tryptophan analogue 5-fluorotryptophan were found to inhibit the growth of wild-type pseudomonas putida. mutants resistant to these analogues were obtained. some of the 5-fluoroindole- and 5-fluorotryptophan-resistant strains exhibit an abnormality in the regulation of certain trp genes. these strains excrete anthranilate when grown in minimal medium in the presence or absence of the inhibitor. in these strains, ...19715573729
induction specificity and catabolite repression of the early enzymes in camphor degradation by pseudomonas putida.the ability of bornane and substituted bornanes to induce the early enzymes for d(+)-camphor degradation and control of these enzymes by catabolite repression were studied in a strain of a pseudomonas putida. bornane and 20 substituted bornane compounds showed induction. of these 21 compounds, bornane and 8 of the substituted bornanes provided induction without supporting growth. oxygen, but not nitrogen, enhanced the inductive potency of the unsubstituted bornane ring. all bornanedione isomers ...19715573731
the metabolism of nicotinic acid. i. purification and properties of 2,5-dihydroxypyridine oxygenase from pseudomonas putida n-9. 19715578917
photoactivation of urocanase in pseudomonas putida. purification of inactive enzyme. 19715580658
sedimentation analysis of pseudomonas putida a.3.12 bacteriophage gh-1 deoxyribonucleic acid. 19675621492
enzymes of the tryptophan synthetic pathway in pseudomonas putida.the first four enzymatic activities of the tryptophan synthetic pathway in pseudomonas putida were found on separate molecules. gel filtration and density gradient centrifugation experiments did not disclose any associations or aggregations among them. these findings contrast with the situation found in the enteric bacteria, where the first two activities are found in an aggregate and the third and fourth are catalyzed by a single enzyme. tryptophan synthetase, the last enzyme of the pathway, co ...19685636809
transducing phage for pseudomonas putida. 19685641761
regulation of histidine catabolism by succinate in pseudomonas putida.the regulation of the histidine-degrading pathway is known to involve induction and repression. our studies have shown that succinate may control the histidine-degrading pathway by sequential negative feedback inhibition. succinate inhibited urocanase, and urocanate in turn inhibited histidase. crude preparations of the two enzymes were made from pseudomonas putida grown on l-histidine. succinate was a competitive inhibitor of urocanase (k(i), 1.8 mm). lactate, pyruvate, alpha-ketoglutarate, and ...19685674054
metabolism of beta-methylaspartate by a pseudomonad.a bacterium was isolated from soil which utilizes threo-beta-methyl-l-aspartate, certain other amino acids, and a variety of organic substances as single energy sources. it is, or closely resembles, pseudomonas putida biotype b. the ability of this organism to rapidly decompose such amino acids is dependent on inducible enzyme systems. dialyzed cell-free extracts of this bacterium metabolize beta-methylaspartate only when catalytic amounts of alpha-ketoglutarate, or pyruvate, and pyridoxal phosp ...19685724974
fine structure mapping of the tryptophan genes in pseudomonas putida. 19685731751
inducible degradation of hydroxyproline in pseudomonas putida: pathway regulation and hydroxyproline uptake.studies in pseudomonas putida of the inducible degradation of hydroxyproline to alpha-ketoglutarate have indicated that either of the two epimers, hydroxy-l-proline or allohydroxy-d-proline, acts as an inducer of all the pathway enzymes. in a mutant lacking the first enzyme of the sequence, hydroxyproline-2-epimerase, which interconverts these two hydroxyproline epimers, either epimer is still equally active as an inducer of the remaining three enzymes, suggesting that each epimer has intrinsic ...19695764334
conversion of l-hydroxyproline to glutamate by extracts of strains of pseudomonas convexa and pseudomonas fluorescens. 19695766672
autonomous replication of a defective transducing phage in pseudomonas putida. 19695784055
isolation of spontaneous mutant strains of pseudomonas putida. 19695796751
crystallization and some properties of 3,4-dihydroxyphenylacetate 2,3-oxygenase from pseudomonas ovalis. 19655857427
[investigations on the oxidative nicotine reduction by pseudomonas convexa(strain 22c)]. 19655899300
the conversion of catechol and protocatechuate to beta-ketoadipate by pseudomonas putida. 19665916391
the conversion of catechol and protocatechuate to beta-ketoadipate by pseudomonas putida. ii. enzymes of the protocatechuate pathway. 19665916392
the conversion of catechol and protocatechuate to beta-ketoadipate by pseudomonas putida. iv. regulation. 19665916393
density heterogeneity in pseudomonas putida bacteriophage preparations. 19665926748
synthesis of the enzymes of the mandelate pathway by pseudomonas putida. i. synthesis of enzymes by the wild type.hegeman, g. d. (university of california, berkeley). synthesis of the enzymes of the mandelate pathway by pseudomonas putida. i. synthesis of enzymes by the wild type. j. bacteriol. 91:1140-1154. 1966.-the control of synthesis of the five enzymes responsible for the conversion of d(-)-mandelate to benzoate by pseudomonas putida was investigated. the first three compounds occurring in the pathway, d(-)-mandelate, l(+)-mandelate, and benzoylformate, are equipotent inducers of all five enzymes. a n ...19665929747
synthesis of the enzymes of the mandelate pathway by pseudomonas putida. ii. isolation and properties of blocked mutants.hegeman, g. d. (university of california, berkeley). synthesis of the enzymes of the mandelate pathway by pseudomonas putida. ii. isolation and properties of blocked mutants. j. bacteriol. 91:1155-1160. 1966.-mutants of pseudomonas putida blocked in early reactions of the pathway for oxidation of d-mandelate were isolated and partially characterized. the specific genetic lesions in these mutants made normal inducer-metabolites of the pathway nonmetabolizable. under the conditions of gratuitous e ...19665929748
synthesis of the enzymes of the mandelate pathway by pseudomonas putida. 3. isolation and properties of constitutive mutants.hegeman, g. d. (university of california, berkeley). synthesis of the enzymes of the mandelate pathway by pseudomonas putida. iii. isolation and properties of constitutive mutants. j. bacteriol. 91:1161-1167. 1966.-mutants of pseudomonas putida constitutive for the synthesis of l(+)-mandelate dehydrogenase were obtained after mandelate- or benzoylformate-limited growth in a chemostat. when grown in media noninducing for the wild type, the mutants are capable of coordinate, constitutive synthesis ...19665929749
characterization of bacteriophage gh-1 for pseudomonas putida.lee, lucy f. (michigan state university, east lansing), and j. a. boezi. characterization of bacteriophage gh-1 for pseudomonas putida. j. bacteriol. 92:1821-1827. 1966.-bacteriophage gh-1 of pseudomonas putida a.3.12 was isolated and purified by differential centrifugation and diethylaminoethyl (deae) cellulose chromatography. an electron micrograph of the phage stained with uranyl acetate revealed a regular hexagonal outline about 50 mmu across with a short wedge-shaped tail attached at one co ...19665958111
cofactor requirements of the l-malate dehydrogenase of pseudomonas ovalis chester.1. the l-malate dehydrogenase of pseudomonas ovalis chester, which is independent of nicotinamide nucleotides and which is structurally and functionally bound to the cell-wall membrane, has been prepared in a soluble form and partially purified. 2. the purified dehydrogenase exhibits a triple cofactor requirement for fad, quinone and phospholipid, and in the presence of these cofactors can utilize 2,6-dichlorophenol-indophenol as hydrogen acceptor. 3. the formation of reduced forms of fad was no ...19665966284
a phage-initiated polysaccharide depolymerase in pseudomonas putida. 19676028945
two new metabolites, 2- nonaprenylphenol and 2-nonaprenyl-3-methyl-6-methoxy-1, 4-benzoquinone, from pseudomonas ovalis. 19676040283
computer simulation of fermentation systems.results of batch fermentation of gluconic acid by pseudomonas ovalis were graphically analyzed to obtain a kinetic model to represent the data. since gluconic acid was produced by the hydrolysis of a lactone intermediate, the model was necessarily represented by a set of kinetic equations. a computer simulation technique involving the use of the midas program was developed to solve the system of nonlinear equations and to check the appropriateness of the model. since the maximal specific growth ...19676049291
metabolism of pipecolic acid in a pseudomonas species. v. pipecolate oxidase and dehydrogenase.oxidation of pipecolate to delta(1)-piperideine-6-carboxylate is catalyzed by pipecolate oxidase, an inducible, membrane-bound dehydrogenase associated with the electron transport components of pseudomonas putida p2. from the oxidase, we obtained a smaller particle containing flavine adenine dinucleotide (fad) and cytochrome b, but no longer able to catalyze electron transfer to oxygen or to cytochrome c. certain properties of this l-pipecolate dehydrogenase, an fad-flavoprotein, are reported.19676051341
cold-sensitive mutation of pseudomonas putida affecting enzyme synthesis at low temperature.a cold-sensitive mutant of pseudomonas putida has been isolated which grows normally at 30 c but is unable to grow on mandelate as a source of carbon at 15 c. the mutation results in the inability of the strain to carry out the reaction catalyzed by cis,cis-muconate lactonizing enzyme at low temperature and must lie in the structural gene for that enzyme, because the mutant enzyme produced at 30 c shows altered thermal stability. the mutant enzyme is not intrinsically cold-labile, nor is it cold ...19676074402
stereochemistry of 1-(4'-hydroxyphenyl)ethanol produced by hydroxylation of 4-ethylphenol by p-cresol methylhydroxylase.enzymic hydroxylation of 4-ethylphenol by (a) pseudomonas putida and (b) highly purified p-cresol methylhydroxylase gave optically active 1-(4'-hydroxyphenyl)-ethanol. the products were transformed into the phenolic methyl ethers and shown to contain 69.5% and 65.6%, respectively, of the (s)-(-)-isomer. the stereochemistry of the reaction is discussed in terms of three distinct steps occurring at the active site of the enzyme.19846083780
[introduction of the hybrid plasmid rp4::d3112 into pseudomonas putida cells requires the presence of specific mutation in the phage genome].the wild type of d3112, a transposable phage of pseudomonas aeruginosa can not be introduced as a portion of the hybrid plasmid rp4::d3112 into pseudomonas putida cells. it is only possible when phage d3112 carries mutations designated lpc (lethal for p. putida and escherichia coli). analysis of heteroduplex molecules between dnas of phages d3112w+ and d3112lpc demonstrated the absence of nonhomology regions, which suggests that lpc is a point mutation. the lpc2 mutation was located within the i ...19846086454
novel system for recognizing and eliminating foreign dna in pseudomonas putida.derivatives of pqsr49 (r1162::tn1) containing cloned fragments of escherichia coli chromosomal dna are stable in e. coli but unstable in pseudomonas putida. similar derivatives containing p. putida chromosomal dna are stable in both species. instability is a consequence of plasmid loss during growth and is not due to death or inhibition of growth of plasmid-containing cells. average copy numbers per cell of the unstable hybrid plasmids are similar to that of pqsr49, indicating that instability i ...19846086582
sequence of the small subunit of yeast carbamyl phosphate synthetase and identification of its catalytic domain.the yeast gene cpa1 coding for the small subunit of arginine-specific carbamyl phosphate synthetase has been cloned by complementation of a cpa 1 mutant with a plasmid library of total yeast chromosomal dna. two of the plasmids, pjl113/st4 and pjl113/st15, contain dna inserts in opposite orientations with overlapping sequences of 2.6 kilobases. the nucleotide sequence of a 2.2-kilobase region of the dna insert carrying the cpa1 gene has been determined. the cpa1 gene has been identified to be 12 ...19846086650
transposon mutagenesis analysis of meta-cleavage pathway operon genes of the tol plasmid of pseudomonas putida mt-2.hybrid plasmids containing the regulated meta-cleavage pathway operon of tol plasmid pwwo were mutagenized with transposon tn1000 or tn5. the resulting insertion mutant plasmids were examined for their ability to express eight of the catabolic enzymes in escherichia coli. the physical locations of the insertions in each of 28 tn1000 and 5 tn5 derivative plasmids were determined by restriction endonuclease cleavage analysis. this information permitted the construction of a precise physical and ge ...19846090417
genetic and physical analyses of caulobacter crescentus trp genes.caulobacter crescentus trp mutants were identified from a collection of auxotrophs. precursor feeding experiments, accumulation studies, and complementation experiments resulted in the identification of six genes corresponding to trpa, trpb, trpc, trpd, trpe, and trpf. genetic mapping experiments demonstrated that the trp genes were in two clusters, trpcde and trpfba, and a 5.4-kilobase restriction fragment from the c. crescentus chromosome was isolated that contained the trpfba gene cluster. co ...19846090420
nucleotide sequence of the promoter region of the xyldegf operon on tol plasmid of pseudomonas putida.the transcription initiation site of the xyldegf operon on the tol plasmid of pseudomonas putida mt-2 was determined in p. putida and in escherichia coli by s1 nuclease and reverse transcriptase mapping. the induced synthesis of mrna started at the same start point in both p. putida and e. coli, although the amount of mrna in e. coli cells was less than that in p. putida. the nucleotide sequence of the region surrounding the start point was also determined. the ribosome-binding site (rbs) comple ...19846092237
transcription of the tol plasmid toluate catabolic pathway operon of pseudomonas putida is determined by a pair of co-ordinately and positively regulated overlapping promoters.expression of the meta-cleavage pathway operon of tol plasmid pww0 of pseudomonas putida is positively regulated by the xyls gene product. we have sequenced the promoter region of this operon and localized the transcription initiation sites. two overlapping promoters, designated pm1 and pm2, are responsible for the positively regulated expression of the meta-pathway operon. mutants of p. putida were isolated that expressed the meta-cleavage pathway operon constitutively. several plasmid-located ...19846096122
purification of glutamine synthetase from a variety of bacteria.we have developed two procedures which allow the very rapid purification of glutamine synthetase (gs) from a diverse variety of bacteria. the first procedure, based upon differential sedimentation, depends upon the association of gs with deoxyribonucleic acid in cell extracts. the second procedure, derived from the method of c. gross et al (j. bacteriol. 128:382-389, 1976) for purifying ribonucleic acid polymerase by polyethylene glycol (peg) precipitation, enabled us to obtain high yields of gs ...19806102984
the conformation of catalytically functional schiff's bases: the pyruvate--lysine ketimine of 2-keto-3-deoxygluconate-6-phosphate aldolase of pseudomonas putida. 19806109521
substrate-mediated inactivation of urocanase from pseudomonas putida. evidence for an essential sulfhydryl group.incubation of urocanase from pseudomonas putida with either its substrate, urocanic acid, or product, 4'(5')-imidazolone-5'(4')-propionic acid, resulted in an oxygen-dependent inhibition of enzyme activity. coincident with the inactivation was the stoichiometric incorporation of radioactivity from [14c]urocanate into the protein. nad+ which is required for activity or urocanase was not directly involved in the inactivation process. the inactivation of urocanase was irreversible, could be partial ...19806109547
mechanism of urocanase as studied by deuterium isotope effects and labeling patterns.nicotinamide adenine dinucleotide (nad) dependent urocanase (4'-imidazolone-5'-propionate hydro-lyase, ec from pseudomonas putida was found to catalyze an exchange reaction between solvent and the 4'-hydrogen of urocanate or imidazolepropionate at a rate faster than that of overall deuterium was compared to unlabeled urocanate as a substrate, no isotope rate effect was noted. for examination of the possibility of an nad+-mediated intramolecular hydride transfer of the 4'-hydrogen to a ...19816110440
glutamine synthetase of pseudomonads: some biochemical and physicochemical properties.the glutamine synthetases from several pseudomonas species were purified to homogeneity, and their properties were compared with those reported for the enzymes from escherichia coli and other gram-negative bacteria. the glutamine synthetase from pseudomonas fluorescens was unique because it was nearly precipitated quantitatively as a homogeneous protein during dialysis of partially purified preparations against buffer containing 10 mm imidazole (ph 7.0) and 10 mm mncl2. the glutamine synthetases ...19816111557
roles of cysteine sulfinate and transaminase on in vitro dark reversion of urocanase in pseudomonas putida.urocanase is inactivated in intact cells of pseudomonas putida and photoactivated by brief exposure of the cells to the uv radiation in sunlight. the dark reversion (inactivation) in vitro is explained by the formation of a sulfite-nad adduct. our objective was to investigate the dark reversion in vivo. various compounds were added to p. putida cells, and the reversion was measured, after sonication, by comparison of the activity before and after uv irradiation. sulfite, cysteine sulfinate, and ...19826124532
identification of the enol tautomer of imidazolone propionate as the urocanase reaction product.a fluorescent product was transiently formed during catalysis by urocanase from pseudomonas putida. the fluorophore showed an emission maximum at 430 nm when excited at 330 nm, essentially identical to that exhibited by the enol tautomer of imidazolone propionate. the keto isomer was not fluorescent under these conditions. in aqueous acid solutions where imidazolone propionate is relatively stable, an equilibrium mixture of tautomeric forms contained approximately 1% of the enol isomer. in ethan ...19836131645
the tol plasmid is naturally derepressed for transfer.pseudomonas putida mt-2, formerly known as pseudomonas arvilla mt-2, which carries the wild-type tol plasmid, and p. putida strain ac37 carrying tol, were completely lysed by the pilus-adsorbing plasmid-specific bacteriophages pr4 and prd1. pseudomonas putida strain pps388, also harbouring the plasmid, was not lysed. in a p. putida mt-2 host, tol transferred 18-fold better on a surface (2.5 x 10(-1) transconjugants per donor h-1) than in liquid; when p. putida pps388 was the host, however, a fre ...19826134782
the iron content of iron superoxide dismutase: determination by anomalous scattering.the number of iron atoms in the dimeric iron-containing superoxide dismutase from pseudomonas ovalis and their atomic positions have been determined directly from anomalous scattering measurements on crystals of the native enzyme. to resolve the long-standing question of the total amount of iron per molecule for this class of dismutase, the occupancy of each site was refined against the measured bijvoet differences. the enzyme is a symmetrical dimer with one iron site in each subunit. the iron p ...19836135208
resolution of 5-oxo-l-prolinase into a 5-oxo-l-proline-dependent atpase and a coupling protein.5-oxo-l-prolinase catalyzes a reaction in which the endergonic cleavage of 5-oxo-l-proline to form l-glutamate is coupled to the exergonic cleavage of atp to adp and pi. in the present research, the enzyme present in a strain of pseudomonas putida isolated from soil by enrichment culture was found to be composed of two protein components. neither component alone could catalyze the 5-oxoprolinase reaction, but the reaction was effectively catalyzed when they were mixed. one component (a) exhibite ...19846145710
pseudomonas putida in transfused blood. 19846145996
similar structures in gamma-carboxymuconolactone decarboxylase and beta-ketoadipate succinyl coenzyme a transferase.gamma-carboxymuconolactone decarboxylase (ec and beta-ketoadipate succinyl coenzyme a transferase (ec mediate different steps in the beta-ketoadipate pathway. antisera prepared against the pseudomonas putida transferase cross-reacted immunologically with the decarboxylase from the same organism. the transferase is formed by association of two nonidentical protein subunits. the nh2-terminal amino acid sequences of the two nonidentical transferase subunits resembled each other a ...19826172419
mutations affecting lipoamide dehydrogenases of pseudomonas putida.pseudomonas putida grown on valine produces two lipoamide dehydrogenases, lpd-glu (mr, 56,000 and lpd-val (mr, 49,000). the 49,000-dalton protein is used by p. putida for branched-chain keto acid dehydrogenase, whereas the 56,000-dalton protein is presumably used for pyruvate and 2-ketoglutarate dehydrogenases. the objective of this study was to isolate and characterize mutants of p. putida with mutations affecting lipoamide dehydrogenases in order to study the relationship of these two proteins ...19836185468
attachment of diaminopimelic acid to bdelloplast peptidoglycan during intraperiplasmic growth of bdellovibrio bacteriovorus early event in the predatory lifestyle of bdellovibrio bacteriovorus 109j is the attachment of diaminopimelic acid (dap) to the peptidoglycan of its prey. attachment occurs over the first 60 min of the growth cycle and is mediated by an extracellular activity(s) produced by the bdellovibrio. some 40,000 dap residues are incorporated into the escherichia coli bdelloplast wall, amounting to ca. 2 to 3% of the total initial dap content of its prey cells. incorporation of dap occurs when e. coli, ...19846202674
the use of mudlac transposons as tools for vital staining to visualize clonal and non-clonal patterns of organization in bacterial growth on agar surfaces.when a histochemical stain for beta-galactosidase activity is applied to growth of gram-negative bacteria on agar medium, the pigmentation is non-uniform and capable of revealing internal colony organization into different cell types. use of an escherichia coli strain with a thermosensitive lac repressor indicates that colonies expand by addition of new cells at the periphery and that older cells which have synthesized beta-galactosidase early in development remain in the centre. mixed inocula o ...19846206197
escherichia coli and pseudomonas putida rna polymerases display identical contacts with promoters.methylation protection experiments with four promoters (p1 and p2 of the pbr322 plasmid, lacuv5 and lambda p0) have shown that the rna polymerases from escherichia coli and pseudomonas putida, while differing in the primary structure of the subunits involved in dna binding, display identical patterns of dna contacts. nor do these enzymes differ in covalent cross-linking patterns with a partially apurinized promoter. we conclude that the two rna polymerases have very similar structures of dna bin ...19846236350
expression of the lactose transposon tn951 in escherichia coli, proteus and pseudomonas.the control of beta-galactosidase specified by the lactose transposon tn951 (inserted into rp1 to give pgc9114) has been studied in escherichia coli k12, proteus mirabilis, pseudomonas aeruginosa and pseudomonas putida; in the first two species comparison could be made with flac. in e. coli k12, the tn951 and chromosomally encoded enzymes showed marked qualitative differences in regulatio, the former giving a substantially lower maximum induced level and induction ratio. several parameters were ...19806251161
properties of six pesticide degradation plasmids isolated from alcaligenes paradoxus and alcaligenes eutrophus.biophysical and genetic properties of six independently isolated plasmids encoding the degradation of the herbicides 2,4-dichlorophenoxyacetic acid and 4-chloro-2-methylphenoxyacetic acid are described. four of the plasmids, pjp3, pjp4, pjp5, and pjp7, had molecular masses of 51 megadaltons, belonged to the incp1 incompatibility group, and transferred freely to strains of escherichia coli, rhodopseudomonas sphaeroides, rhizobium sp., agrobacterium tumefaciens, pseudomonas putida, pseudomonas flu ...19816257648
cyclic adenosine 3',5'-monophosphate levels in pseudomonas putida and pseudomonas aeruginosa during induction and carbon catabolite repression of histidase synthesis.inducibility of histidase (histidine ammonia-lyase, ec in pseudomonas putida and pseudomonas aeruginosa was observed to be strongly affected by succinate-provoked catabolite repression, but this did not occur as a consequence of reduced intracellular cyclic adenosine 3',5'-monophosphate levels, and repression could not be alleviated by exogenously added cyclic adenosine 3,'5'-monophosphate. milder repression of histidase by lactate was also not reversed by the addition of cyclic adenosi ...19816259129
[plasmid characteristics of naphthalene and salicylate biodegradation in pseudomonas putida].the object of this work was to study the physico-chemical and biological properties of dnas of the biodegradation plasmids nah and sal. a comparative analysis of the physico-chemical parameters for these dnas made it possible to detect a number of identical properties in them: the same sedimentation profile for covalently-closed circular dna forms, 68--70 s; the molecular weight of ca. 50 md; a roughly equal number of fragments (up to 23) was found when the dnas of nah and sal were restricted by ...19806259498
in vitro antimicrobial activity of ceftizoxime against glucose-nonfermentative gram-negative rods.ceftizoxime, a new cephalosporin, was active against pseudomonas cepacia, flavobacterium meningosepticum, alcaligenes faecalis, and acinetobacter calcoaceticus and was more potent against pseudomonas aeruginosa and pseudomonas putida than was carbenicillin.19816269480
the respiratory system of pseudomonas putida: participation of cytochromes in electron transport. 19816269495
molecular cloning of gene xyls of the tol plasmid: evidence for positive regulation of the xyldegf operon by xyls.the xyldegf operon and the regulatory gene xyls of the tol plasmid found in pseudomonas putida mt-2 were cloned onto escherichia coli vector plasmids. a 9.5-kilobase fragment, derived from the tol segment of ptn2 deoxyribonucleic acid, carried the xyl genes d, e, g, and f, which encode toluate oxygenase, catechol 2,3-oxygenase, 2-hydroxymuconic semialdehyde dehydrogenase, and 2-hydroxymuconic semialdehyde hydrolase, respectively. the enzymes were noninducible unless a 3-kilobase psti fragment, d ...19816271729
an electron-spin-resonance study on the redox-active centers of the 4-methoxybenzoate monooxygenase from pseudomonas putida. 19816273164
purification and properties of protocatechuate 3,4-dioxygenase from pseudomonas putida. a new iron to subunit stoichiometry.protocatechuate dioxygenase has been isolated from pseudomonas putida. this new species of protocatechuate dioxygenase has been characterized and compared with the enzyme from pseudomonas aeruginosa. the enzyme reported here has visible absorption, circular dichroism, electron paramagnetic resonance, and raman spectroscopic properties virtually identical to those for protocatechuate dioxygenase from p. aeruginosa. however, the molecular weight and iron:subunit stoichiometry differ. protocatechua ...19816273403
mössbauer studies on the active fe ... [2fe-2s] site of putidamonooxin, its electron transport and dioxygen activation mechanism.putidamonooxin, the oxygenase of a 4-methoxybenzoate monooxygenase enzyme system, catalyzes the oxidative o-demethylation of the substrate 4-methoxybenzoate in conjunction with the nadh:putidamonooxin oxidoreductase. putidamonooxin is a conjugated iron-sulfur protein which needs iron ions as cofactors for its enzymatic activity. putiamonooxin was isolated from pseudomonas putida, which was grown on a 57fe-enriched culture medium. thus putidamonooxin was enriched in vivo with 57fe up to about 80% ...19816276173
plasmid gene organization: naphthalene/salicylate oxidation.genes for naphthalene metabolism are localized on nah7, an 83-kilobase (kb) plasmid, in two gene clusters under salicylate control. polar mutations formed by insertion of the transposon tn5 permit detection of the transcription direction and the gene organization within two approximately 10-kb dna segments separated by a approximately 7-kb regulatory gene region. the gene cluster specifying conversion of naphthalene to salicylate lies near the left initiation of a 25-kb dna fragment a released b ...19826278499
broad-host-range incp-4 plasmid r1162: effects of deletions and insertions on plasmid maintenance and host range.r1162 is an 8.7-kilobase (kb) broad-host-range replicon encoding resistance to streptomycin and sulfa drugs. in vitro deletion of 1.8-kb dna between coordinates 3.0 and 5.3 kb did not affect plasmid maintenance, but a tn1 insertion at coordinate 6.3 kb led to a recessive defect in plasmid maintenance. the only cis-acting region necessary for plasmid replication appears to lie between the tn1 insertion at coordinate 6.3 kb and a second tn1 insertion at coordinate 6.5 kb. all r1162 sequences betwe ...19826288654
spontaneous deletions in the tol plasmid pww20 which give rise to the b3 regulatory mutants of pseudomonas putida mt20.the size of the tol plasmid pww20 from pseudomonas putida mt20, as measured by analysis of agarose electrophoresis gels after restriction endonuclease hydrolysis, was 270-280 kilobase pairs (kb). during growth on benzoate, mt20 segregates strains carrying mutations in the plasmid regulatory gene xyls; these so-called b3 strains retain the ability to grow on m-xylene (mxy+) but do not grow on its metabolite m-toluate (mtol-) and have also lost the ability to transfer the plasmid (tra-). analysis ...19826288840
identification of chromosomally integrated tol dna in cured derivatives of pseudomonas putida paw1.some plasmid-free tol- strains derived from pseudomonas putida paw1 (which carries the tol plasmid pww0) have a segment of tol dna located chromosomally. of three independently isolated strains, paw86 had an integrated tol segment of 16 kilobases and paw85 had two copies of this segment in different chromosomal locations, whereas the chromosomal dna of paw82 showed no homology with the tol plasmid. in cultures of the parental strain, it appears that a 56-kilobase tol dna segment is located chrom ...19826290457
integration of r91-5::tn501 into the pseudomonas putida ppn chromosome and genetic circularity of the chromosomal map.derivatives of the pseudomonas aeruginosa plasmid r91-5, loaded with the transposon tn501, were transferred to p. putida ppn. over 90% of exconjugants, which arose at a frequency of ca. 10(-6) per donor cell, exhibited high-frequency (greater than 10(-2) per donor cell) polarized transfer of chromosomal markers. in one instance it was demonstrated by transduction that the plasmid had been inserted into a gene required for serine biosynthesis. the integrated nature of the plasmid in this and othe ...19836294058
cloning and expression in escherichia coli of the naphthalene degradation genes from plasmid nah7.the genes encoding the enzymes responsible for conversion of naphthalene to 2-hydroxymuconic acid (naha through nahi) are contained on a 25-kilobase ecori fragment of an 85-kilobase nah plasmid of pseudomonas putida. these genes were cloned into the plasmid vectors pbr322 and rsf1010 to obtain the recombinant plasmids pkgx505 and pkgx511, respectively. to facilitate cloning and analysis, an nah7 plasmid containing a tn5 transposon in the salicylate hydroxylase gene (nahg) was used to derive the ...19836296054
proton coupling in the ligand-binding reaction of ferric cytochrome p-450 from pseudomonas putida.effects of ph on the ligand-binding reactions of ferric heme in cytochrome p-450 from pseudomonas putida (camphor 5-monooxygenase, ec were studied by using cyanide, n-methylimidazole, pyridine, and ethylisocyanide as ligands. in all cases, affinity of the ferric heme for the ligand was found to increase as ph of the medium was raised from around 6 to 9. depending on the ligand, the increase was 10- to 1000-fold and the shapes of their ph-affinity curves were remarkably different. anal ...19836301381
structural characteristics of cytochrome p-450. possible location of the heme-binding cysteine in determined amino-acid analyses were made of the complete amino-acid sequences of two p-450 species, the phenobarbital-inducible major p-450 of rat liver microsomes (p-450pb) and camphor-hydroxylating p-450 of pseudomonas putida (p-450cam). statistically significant homology was recognized between the two p-450 sequences, but these sequences were not related to those of other groups of hemoproteins, such as hemoglobins, peroxidases, and cytochrome c's and b's. two highly homologous regions, hr1 and hr2, ...19836307988
redox potential of the cytochrome c in the flavocytochrome p-cresol methylhydroxylase.the redox potential of the cytochrome c in 5 flavocytochrome c proteins, all p-cresol methylhydroxylases purified from species of pseudomonas, was measured. all gave similar values ranging from 226-250 mv. two of the enzymes, from pseudomonas putida nc1b 9866 and nc1b 9869, were resolved into their flavoprotein and cytochrome subunits and the redox potentials of the isolated cytochrome c subunits measured. the values for these were 60-70 mv below those for the whole enzymes but, in both cases, r ...19836309572
susceptibility of 324 nonfermentative gram-negative rods to 6 cephalosporins and azthreonam.susceptibility of 324 isolates of nonfermentative gram-negative bacteria to cephalothin, cefamandole, cefoxitin, ceftazidime, cefsulodin, and azthreonam was determined by agar dilution and disc diffusion techniques. with the exception of moraxella species, first- and second-generation cephalosporins were minimally active against nonfermenters tested. cefsulodin and azthreonam were mainly active against pseudomonas aeruginosa. in contrast, ceftazidime and ceftriaxone exhibited wider activity spec ...19836311492
cloning of genes for naphthalene metabolism in pseudomonas putida.plasmid pig7 dna cloned in pseudomonas putida with the broad-host-range vectors prk290 and pkt240 expresses the genes encoding nephthalene oxidation in the presence of the intermediate substrate, salicylate, or the gratuitous inducer, anthranilate. two operons, nahaf and nahgk, cloned from the ecori fragment a (25 kilobases) are under wild-type regulation by the nahr locus. deletion plasmids provide a restriction map of both operons. double transformants containing structural and regulatory cist ...19836311809
plasmid-encoded regulation of colicin e1 gene expression.a plasmid-encoded factor that regulates the expression of the colicin e1 gene was found in molecular cloning experiments. the 2,294-base-pair avaii fragment of the colicin e1 plasmid (cole1) carrying the colicin e1 structural gene and the promoter-operator region had the same information with respect to the repressibility and inducibility of colicin e1 synthesis as the original cole1 plasmid. an operon fusion was constructed between the 204-bp fragment containing the colicin e1 promoter-operator ...19836313603
use of ureidopenicillins for selection of plasmid vector transformants in pseudomonas aeruginosa and pseudomonas putida.broad-host-range plasmids coding for beta-lactamase were successfully selected after transformation of pseudomonas strains. transformants of both pseudomonas aeruginosa and pseudomonas putida containing plasmid pro1614 were isolated in media containing low concentrations of piperacillin. these strains were also susceptible to other ureidopenicillins. similar selections of transformants with carbenicillin, ampicillin, or ticarcillin required high concentrations of antibiotics and yielded backgrou ...19846321446
activity of the hybrid trp-lac (tac) promoter of escherichia coli in pseudomonas putida. construction of broad-host-range, controlled-expression vectors.a broad-host-range vector, pkt240, containing the structural gene (aph) for aminoglycoside phosphotransferase (aph), without promoter, has been constructed. insertion of dna fragments carrying promoters upstream of aph gene into the unique ecori site of this vector results in the expression of the aph gene and consequently the resistance of the host cells to streptomycin. the new vector has been used to show that the hybrid trp-lac (tac) promoter and the promoter of the laciq gene of escherichia ...19836323265
nucleotide sequence surrounding transcription initiation site of xylabc operon on tol plasmid of pseudomonas putida.the xylabc operon on the tol plasmid directs the synthesis of enzymes for conversion of toluene to benzoate and is positively controlled by the regulatory gene xylr. in the study here the nucleotide sequence was determined for the regulatory region of this operon. the in vivo transcription initiation site of the operon was determined by s1 nuclease and reverse transcriptase mapping. rna was prepared from m-methylbenzyl alcohol-induced cells of pseudomonas putida and escherichia coli carrying ptn ...19846324212
an investigation of the iron-sulphur proteins of benzene dioxygenase from pseudomonas putida by electron-spin-resonance spectroscopy.benzene dioxygenase from pseudomonas putida comprises three components, namely a flavoprotein (nadh:ferredoxin oxidoreductase; mr 81000), an intermediate electron-transfer protein, or ferredoxin (mr 12000) with a [2fe-2s] cluster, and a terminal dioxygenase containing two [2fe-2s] iron-sulphur clusters (mr 215000), which requires two additional fe2+ atoms/molecule for oxygenase activity. the ferredoxin and the dioxygenase give e.s.r. signals in the reduced state with rhombic symmetry and average ...19846324743
enzyme recruitment in vitro: use of cloned genes to extend the range of haloaromatics degraded by pseudomonas sp. strain b13.dna fragments containing the xyld and xyll genes of tol plasmid pww0 -161 of pseudomonas putida, which code for the catabolic enzymes toluate 1,2-dioxygenase and dihydrodihydroxybenzoic acid dehydrogenase, respectively, and the nahg gene of the nah plasmid nah7 , which codes for salicylate hydroxylase, were cloned in pbr322 vector plasmid. deletion and insertion mutagenesis were used to localize these genes with respect to crucial endonuclease cleavage sites. the pbr322-based plasmids were ligat ...19846327621
isolation and expression of rhizobium japonicum cloned dna encoding an early soybean nodulation function.a first visible step in the nodulation of legumes by rhizobium spp. is the deformation and curling of root hairs. we have identified and cloned dna sequences encoding this function from two strains of rhizobium japonicum (usda 122 and usda 110) with a weakly homologous probe from rhizobium meliloti. root hair curling encoded by the cloned dna fragments was examined on soybeans (glycine soja ) after conjugative transfer of these sequences in broad-host-range vectors to various bacterial genera. p ...19846327649
transposon tn5 encodes streptomycin resistance in nonenteric bacteria.strains of caulobacter crescentus, pseudomonas putida, acinetobacter calcoaceticus, rhizobium meliloti, and rhodopseudomonas sphaeroides carrying the kanamycin resistance-encoding transposon tn5 were 15 to 500 times more resistant to streptomycin than transposon-free strains. the streptomycin resistance determinant, which is separable from the kanamycin resistance determinant of tn5, was not expressed in escherichia coli or klebsiella aerogenes.19846330041
stereospecificity in meta-fission catabolic pathways.the stereospecificities of 2-keto-4-pentenoate hydratase and 4-hydroxy-2-ketovalerate aldolase were studied in escherichia coli, pseudomonas putida, and acinetobacter sp. hydration was stereospecific in all three; however, only p. putida and acinetobacter sp. showed stereospecificity in their aldolase reactions.19836345511
replication of derivatives of the broad host range plasmid rk2 in two distantly related bacteria.a 0.7-kb segment of the broad host range plasmid rk2 containing the replication origin of this plasmid will replicate in escherichia coli and pseudomonas putida when this segment is joined to a 1.8-kb region of rk2 designated traa*. the presence of another region of rk2, designated trfb, that previously was implicated in rk2 replication had no effect on the maintenance of the rk2 trfa*-oriv replicon in these two organisms. these observations indicate a requirement for a minimal account of inform ...19836346358
expression of naphthalene oxidation genes in escherichia coli results in the biosynthesis of indigo.a fragment of plasmid nah7 from pseudomonas putida ppg7 has been cloned and expressed in escherichia coli hb101. growth of the recombinant escherichia coli in nutrient medium results in the formation of indigo. the production of this dye is increased in the presence of tryptophan or indole. several bacteria that oxidize aromatic hydrocarbons to cis-dihydrodiols also oxidize indole to indigo. the results suggest that indigo formation is due to the combined activities of tryptophanase and naphthal ...19836353574
molecular cloning of the pseudomonas carboxypeptidase g2 gene and its expression in escherichia coli and pseudomonas putida.the gene coding for carboxypeptidase g2 was cloned from pseudomonas sp. strain rs-16 into escherichia coli w5445 by inserting sau3a-generated dna fragments into the bamhi site of pbr322. the plasmid isolated, pnm1, was restriction mapped, and the position of the gene on the 5.8-megadalton insert was pinpointed by subcloning. the expression of carboxypeptidase in e. coli was 100-fold lower than in the pseudomonas sp. strain. when the cloned gene was subcloned into the pseudomonas vector pkt230 an ...19836358192
permeability of the boundary layers of bdellovibrio bacteriovorus 109j and its bdelloplasts to small hydrophilic molecules.measurements of the sucrose-permeable and -impermeable volumes during bdellovibrio bacteriovorus attack on escherichia coli or pseudomonas putida showed that the volume of the bdelloplast increased over that of the substrate cell. although the pattern of the increase differed with the two organisms, the volumes reached maximum at about 60 min into the bdellovibrio growth cycle. by this time, the cytoplasmic membranes of the attacked cells were completely permeable to sucrose. the kinetics of inc ...19846363383
relationship of lipoamide dehydrogenases from pseudomonas putida to other fad-linked dehydrogenases.pseudomonas putida produces two lipoamide dehydrogenases, lpd-glc and lpd-val. lpd-val is specifically required as the lipoamide dehydrogenase of branched-chain keto acid dehydrogenase and lpd-glc fulfills all other requirements for lipoamide dehydrogenase. both proteins are dimers with one fad per subunit. lpd-glc has an absorption maximum at 455 nm, but lpd-val has a maximum at 460 nm. comparison of amino acid compositions revealed that lpd-glc was more closely related to escherichia coli and ...19846373365
expression of the argf gene of pseudomonas aeruginosa in pseudomonas aeruginosa, pseudomonas putida, and escherichia coli.r' plasmids carrying argf genes from pseudomonas aeruginosa strains pao and pac were transferred to pseudomonas putida argf and escherichia coli argf strains. expression in p. putida was similar to that in p. aeruginosa and was repressed by exogenous arginine. expression in e. coli was 2 to 4% of that in p. aeruginosa. exogenous arginine had no effect, and there were no significant differences between argr' and argr strains of e. coli in this respect.19836403512
chromogenic identification of genetic regulatory signals in bacillus subtilis based on expression of a cloned pseudomonas gene.a method to isolate fragments of dna that promote gene expression in bacillus subtilis is described. the system is based on production of catechol 2,3-dioxygenase [cato2ase; catechol:oxygen 2,3-oxidoreductase (decyclizing), ec] encoded by the pseudomonas putida tol plasmid gene xyle. the gene was transferred to ab. subtilis/escherichia coli plasmid vector to construct ptg402. although xyle is functionally expressed in e. coli, cato2ase is not detected in b. subtilis unless a fragment o ...19836405380
[molecular dna-dna hybridization in pseudomonas fluorescens and pseudomonas putida].taxonomic analysis was carried out and dna-dna homology studied in cultures of subgroups formed by numerical classification of 124 pseudomonas fluorescens and p. putida strains. the gc content in the dna of strains belonging to p. fluorescens subgroups varied from 61.2 to 64.5%, and their dna-dna homology with the type strain p. fluorescens atcc 13525 was 24 to 83%. the lowest genome relatedness with the type culture was found in biotype c of p. fluorescens and p. aureofaciens strains. the gc c ...19836413827
[transformation of escherichia coli and different species of pseudomonas by a plasmid dna isolated from pseudomonas testosteroni].pseudomonas testosteroni uses testosterone as sole source of carbon. we were able to isolate an extrachromosomal dna from a strain of pseudomonas testosteroni and to obtain pseudomonas putida and aeruginosa and escherichia coli transformants catabolizing testosterone.19836416627
l-arginine utilization by pseudomonas species.the utilization of arginine was studied in several different pseudomonas species. the arginine decarboxylase and agmatine deiminase pathways were found to be characteristic of pseudomonas species of group i as defined by palleroni et al. (1974). pseudomonas putida strains had three distinct arginine catabolic pathways initiated by arginine decarboxylase, arginine deiminase and arginine oxidase, respectively. the two former routes were also present in p. fluorescens and p. mendocina and in p. aer ...19846423769
purification and properties of amino acid racemase from aeromonas punctata subsp. amino acid racemase, which occurs in the cytoplasmic fraction of aeromonas punctata subsp. caviae, has been purified to homogeneity by the criteria of electrophoresis and ultracentrifugation. the enzyme has a molecular weight of about 80,000 and consists of two subunits identical in molecular weight (about 40,000). the enzyme contains 2 mol of pyridoxal 5'-phosphate per mol of enzyme, and exhibits absorption maxima at 280 nm and 420 nm. the holoenzyme is resolved by dialysis against hydroxyla ...19846427786
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