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specificity of pyoverdine-mediated iron uptake among fluorescent pseudomonas strains.pyoverdine-mediated iron transport was determined for seven fluorescent pseudomonas strains belonging to different species. for all strains, cell or cell outer membrane and iron(iii)-pyoverdine combinations were compared with their homologous counterparts in uptake, binding, and cross-feeding experiments. for four strains (pseudomonas putida atcc 12633, pseudomonas fluorescens w, p. fluorescens atcc 17400, and pseudomonas tolaasii ncppb 2192), the pyoverdine-mediated iron transport appeared to b ...19883170485
[clinical study of cefuzonam in the field of obstetrics and gynecology].cefuzonam (czon, l-105) was used clinically for the treatment of obstetrical and gynecological infections at a dosage of 1 g once or twice daily by intravenous drip infusion. the results obtained are summarized as follows. 1. clinical effects of czon were analyzed in 10 patients, including 5 patients with intrapelvic infections, 3 with intrauterine infections, and 1 each with adnexitis and an external genital infection. excellent responses were observed in 1 patient (11.1%), good responses in 7 ...19883172464
[physical map of the dna of bacteriophage tf of pseudomonas putida].a physical map has been constructed for p. putida bacteriophage tf dna containing single-strand breaks (nicks). localization of cleavage sites for ecori, hindiii, hpai clai, bamhi, sali, xbai and xhoi restriction endonucleases was determined. position of single-strand breaks was mapped by electrophoretic analysis of denatured tf dna and electron microscopy of partially denatured dna samples. the tf genome is characterized by the presence of two classes of nicks differing in the frequency of thei ...19883173374
[purification and properties of two enzymes of meta-cleaving the aromatic ring controlled by the biphenyl biodegradation plasmid pbs 241 from pseudomonas putida].it was shown that two metapyrocatechases (ec 1.13.11.2) function in pseudomonas putida bs893. biphenyl degradative plasmid pbs241 carries the genes of these enzymes. the basic properties of the both enzymes, i. e., mpc1 and mpc2, were investigated. it was found that mpc1 is an enzyme with a molecular mass of 135 kd and has a heterotetrameric subunit structure (alpha 2 beta 2), being made up of two non-identical polypeptides with mr of 34 and 22.5 kd; pi is 5.15, the ph optimum is at 8.0, a tempe ...19883179349
[genes coding for rna-polymerase in bacteria. ii. conservative sites in the central region of the beta-subunit of pseudomonas putida rna-polymerase].sali--l fragment of the p. putida rpobc operon has been sequenced and conservative regions of the central part of the rna-polymerase beta-subunit have been determined. amino and acid residues interacting with zn2+ are postulated.19883190780
[isolation, purification and various properties of l-lysine-2-monooxygenase from pseudomonas putida].isolation and purification of l-lysine-2- monooxygenase from the bacterium pseudomonas species was carried out. the purification procedure included ammonium sulfate fractionation, acid treatment, gel filtration through sephadex g-200 and ion-exchange chromatography on deae-sephadex a-50. such treatment resulted in more than 220-fold purification and 22% yield; the specific activity of the enzyme is 14.6 u/mg. the enzyme spectrum is typical for flavoproteins, with peaks at 275, 386 and 462 nm. at ...19883191192
purification and properties of catechol 1,2-dioxygenase (pyrocatechase) from pseudomonas putida mt-2 in comparison with that from pseudomonas arvilla c-1.catechol 1,2-dioxygenase (pyrocatechase) has been purified to homogeneity from pseudomonas putida mt-2. most properties of this enzyme, such as the absorption spectrum, iron content, ph stability, ph optimum, substrate specificity, km values, and amino acid composition, were similar to those of catechol 1,2-dioxygenase obtained from pseudomonas arvilla c-1 [y. kojima et al. (1967) j. biol. chem. 242, 3270-3278]. these two catechol 1,2-dioxygenases were also found, from the results of ouchterlony ...19883214177
[genes coding for rna polymerase in bacteria. iii. the use of modified sanger's method for sequencing the c-terminal region of rpob gene, n-terminal region of rpoc gene and intercistron region of rna polymerase in pseudomonas putida].the sanger method was modified and the primary structure of the sali-c fragment of the pseudomonas putida rpobc operon was elucidated.19883219133
crystal structure determination, refinement and molecular model of creatine amidinohydrolase from pseudomonas putida.the three-dimensional crystal structure of creatine amidinohydrolase (creatinase ec 3.5.3.3) from pseudomonas putida, a dimeric enzyme with a molecular weight of 97,000, has been determined by multiple isomorphous replacement, averaging over the local dyad and restrained crystallographic refinement at 1.9 a with a crystallographic r-value of 17.7%. the asymmetric unit contains a dimer. the two chemically identical subunits consist of 403 residues each. a subunit is built up of two domains, a sma ...19883221393
4-methoxybenzoate monooxygenase from pseudomonas putida: isolation, biochemical properties, substrate specificity, and reaction mechanisms of the enzyme components. 19883226294
in vivo generation of r68.45-ppgh1 hybrid plasmids conferring a phl+ (meta pathway) phenotype.plasmid ppgh1 originating from pseudomonas putida strain h carries all the genes required for the degradation of phenol (or cresols) via the meta cleavage pathway. besides mobilization of ppgh1 by a plasmid of the incompatibility group p-1, hybrid plasmids conferring the phl+ phenotype could be selected, when r68.45 was the conjugative plasmid. the hybrids contain the complete r68.45 and part of ppgh1. integration of phl-dna of ppgh1 into r68.45 occurred exclusively via the is21 region of r68.45 ...19883226424
degradation of 1,4-naphthoquinones by pseudomonas putida.pseudomonas putida j1 and j2, enriched from soil with juglone, are capable of a total degradation of 1,4-naphthoquinone, 2-hydroxy-1,4-naphthoquinone, and 2-chloro-1,4-naphthoquinone. naphthazerin and plumbagin are only converted into the hydroxyderivatives 2-hydroxynaphthazerin and 3-hydroxyplumbagin, respectively, whereas 2-amino-1,4-naphthoquinone is not attacked at all. the degradation of 1,4-naphthoquinone begins with a hydroxylation of the quinoid ring, yielding 2-hydroxy-1,4-naphthoquinon ...19883228489
cloning, nucleotide sequence and characterization of genes encoding naphthalene dioxygenase of pseudomonas putida strain ncib9816.we have cloned the naphthalene dioxygenase(nd)-coding genes from pseudomonas putida strain ncib9816 based on their ability to convert indole to indigo. the region coding for this enzyme activity was sequenced and three successive open reading frames were found. the corresponding gene products were identified using the t7 polymerase/promoter system. all of them are necessary for the nd activity. a comparison of the nd-coding genes with the ones coding for benzene dioxygenase revealed significant ...19883243438
loss of the toluene-xylene catabolic genes of tol plasmid pww0 during growth of pseudomonas putida on benzoate is due to a selective growth advantage of 'cured' segregants.during growth on benzoate-minimal medium pseudomonas putida mt-2 (paw1) segregates derivative ('cured') strains which have lost the ability to use the pathway encoded by its resident catabolic plasmid pww0. experiments with two plasmids identical to pww0 but each with an insert of tn401, which confers resistance to carbenicillin, suggested that the 'benzoate curing' occurs far more frequently by the specific deletion of the 39 kbp region carrying the catabolic genes than by total plasmid loss. t ...19883246596
[a comparative study of the formation of 2-keto-d-gluconic acid by free and immobilized cells of pseudomonas putida].the effect of glucose, oxygen and 2-keto-d-gluconic acid (2kg) concentrations on the 2kg production by free and immobilized cells of pseudomonas putida was studied. the effect of these factors was found to be similar in case of both free and immobilized cells, but the rate of the 2kg production by the free cells was a little higher as compared to the immobilized cells.19883249741
[purification and properties of beta-ketoadipyl-coenzyme a thiolase from pseudomonas putida]. 19883250096
comparison of the meta pathway operons on nah plasmid pww60-22 and tol plasmid pww53-4 and its evolutionary significance.the regulated meta pathway operon for the catabolism of salicylate on the naphthalene plasmid pww60-22 was cloned into the broad-host-range vector pkt230 on a 17.5 kbp bamhi fragment. the recombinant plasmid conferred the ability to grow on salicylate when mobilized into plasmid-free pseudomonas putida paw130. a detailed restriction map of the insert was derived and the locations of some of the genes were determined by subcloning and assaying for their gene products in escherichia coli and p. pu ...19883254935
in vitro and in vivo antibacterial activities of me1207, a new oral cephalosporin.me1207 (pivaloyloxymethyl ester of me1206) is a new oral cephalosporin. me1206 is (6r,7r)-7-[(z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)- acetamido]-3-[(z)-2-(4-methylthiazol-5-yl)-ethyl]-cephem-4-carboxy lic acid. the susceptibilities of about 1,600 clinical isolates to me1206 were determined by the agar dilution method. me1206 showed a broad spectrum of activity against gram-positive and gram-negative bacteria. me1206 was more active than cefaclor, t-2525, and cefixime against staphylococcus ...19883264132
organization and nucleotide sequence determination of a gene cluster involved in 3-chlorocatechol degradation.three critical enzymes catechol oxygenase ii (chlorocatechol dioxygenase), muconate cycloisomerase ii, and dienelactone hydrolase, are involved in the degradation of chlorocatechols, which are obligatory intermediates in the catabolism of chlorinated aromatic compounds. the organization and complete nucleotide sequence of the genes for these enzymes have been determined on a 4.2-kilobase-pair (kbp) bgl ii fragment cloned from the plasmid pac27, based on the agreement of open reading frame length ...19873299368
maintenance and stability of introduced genotypes in groundwater aquifer material.three indigenous groundwater bacterial strains and pseudomonas putida harboring plasmids tol (pwwo) and rk2 were introduced into experimentally contaminated groundwater aquifer microcosms. maintenance of the introduced genotypes was measured over time by colony hybridization with gene probes of various specificity. on the basis of the results of colony hybridization quantitation of the introduced organisms and genes, all introduced genotypes were stably maintained at approximately 10(5) positive ...19873300546
activation of the xyldlegf promoter of the tol toluene-xylene degradation pathway by overproduction of the xyls regulatory gene product.the xyls regulatory gene of the pseudomonas putida tol plasmid (pwwo) has been cloned under the transcriptional control of the escherichia coli tac promoter in a broad-host-range controlled-expression vector. induction with isopropylthiogalactoside allowed overproduction and characterization of the xyls product by specific interaction with the tol meta-cleavage pathway operator-promoter region (op2) in vivo in e. coli. examination of plasmid-specified polypeptides in e. coli maxicells led to ide ...19873301806
purification and properties of formylglutamate amidohydrolase from pseudomonas putida.formylglutamate amidohydrolase (fgase) catalyzes the terminal reaction in the five-step pathway for histidine utilization in pseudomonas putida. by this action, n-formyl-l-glutamate (fg) is hydrolyzed to produce l-glutamate plus formate. urocanate, the first product in the pathway, induced all five enzymes, but fg was able to induce fgase alone, although less efficiently than urocanate did. this induction by fg resulted in the formation of an fgase with electrophoretic mobility identical to that ...19873308850
molecular analysis of regulatory and structural xyl genes of the tol plasmid pww53-4.pww53-4 is a cointegrate between rp4 and the catabolic plasmid pww53 from pseudomonas putida mt53, which contains 36 kbp of pww53 dna inserted close to the oriv gene of rp4; it encodes the ability to grow on toluene and the xylenes, characteristic of pww53, as well as resistance to tetracycline, kanamycin and carbenicillin, characteristic of rp4. a physical map of the 36 kbp insert of pww53 dna for 11 restriction enzymes is presented, showing that the relative positions of the two xyl operons ar ...19873309179
effect of temperature on the stability of plasmid ptg201 and productivity of xyle gene product in recombinant escherichia coli: development of a two-stage chemostat with free and immobilized cells.the effect of temperature on the stability of ptg201, a plasmid carrying the xyle gene (which encodes catechol 2,3-dioxygenase from pseudomonas putida), and the production of catechol 2,3-dioxygenase in free and immobilized escherichia coli during continuous culture have been studied at various temperatures. immobilization of cells increased the stability of ptg201 considerably, even under conditions when expression of the xyle product was enhanced. since xyle transcription was controlled by the ...19873312486
[molecular genetic organization and origin of plasmid pbs52 with a broad range of bacterial hosts].the data are presented on the localization of genetic determinants of resistance to streptomycin, ampicillin and sulfanilamides on the physical map of conjugative r plasmid pbs52 of 38,000 bp which has a broad bacterial host range and belongs to a new incompatibility group. the plasmid has a natural "polylinker" site (less than 200 bp) containing (in the order of arrangement) the recognition sites for restriction enzymes: bamhi-ecori-psti-ecorv-bglii (pvuii). the comparative analysis shows that ...19873319772
qualitative evidence for expression of klebsiella pneumoniae nif in pseudomonas putida.pseudomonas putida mt20-3 carrying the klebsiella pneumoniae nif plasmids prd1 or pmf250 showed highly o2-sensitive aerobic acetylene reduction on low-n pyruvate or glucose agar. this finding confirms unequivocally that k. pneumoniae nif can be expressed in an obligate aerobe.19873329216
purification and characterization of a bacterial nitrophenol oxygenase which converts ortho-nitrophenol to catechol and nitrite.a nitrophenol oxygenase which stoichiometrically converted ortho-nitrophenol (onp) to catechol and nitrite was isolated from pseudomonas putida b2 and purified. the substrate specificity of the enzyme was broad and included several halogen- and alkyl-substituted onps. the oxygenase consisted of a single polypeptide chain with a molecular weight of 58,000 (determined by gel filtration) or 65,000 (determined on a sodium dodecyl sulfate-polyacrylamide gel). the enzymatic reaction was nadph dependen ...19883350791
[products of biphenyl catabolism by a pseudomonas putida strain carrying the biodegradation plasmid pbs 241]. 19883351100
trichloroethylene metabolism by microorganisms that degrade aromatic compounds.trichloroethylene (tce) was metabolized by the natural microflora of three different environmental water samples when stimulated by the addition of either toluene or phenol. two different strains of pseudomonas putida that degrade toluene by a pathway containing a toluene dioxygenase also metabolized tce. a mutant of one of these strains lacking an active toluene dioxygenase could not degrade tce, but spontaneous revertants for toluene degradation also regained tce-degradative ability. the resul ...19883355147
properties and functions of two succinic-semialdehyde dehydrogenases from pseudomonas putida.two forms of succinic-semialdehyde dehydrogenase have been isolated in pseudomonas putida. the two enzymes could be separated by filtration on sephacryl s-300 and their apparent molecular weights were approx. 200,000 and 100,000. the smaller enzyme, which is induced by growth on 4-hydroxyphenylacetate, has been purified to 88% homogeneity by anion-exchange and affinity chromatography. electrophoresis in sodium dodecyl sulphate gave rise to a molecular weight of 53,000, indicating that the native ...19883355840
primary structure of protein b from pseudomonas putida, member of a new class of 2fe-2s ferredoxins.the primary structure of the 2fe-2s ferredoxin (protein b) from the benzene dioxygenase system of pseudomonas putida strain ncib 12190 was determined by gas-phase sequencing of the protein and its fragments. fast atom bombardment mass spectrometry indicated a molecular mass of 11,860 da. the sequence contained five cysteine residues, four of which would be required to coordinate the iron-sulphur cluster. the amino acid sequence determined in the present study is compared to that of a protein ded ...19883360142
interaction of the protein components of 5-oxoprolinase. substrate-dependent enzyme complex formation.5-oxo-l-prolinase from pseudomonas putida is composed of two reversibly dissociable proteins: component a catalyzes 5-oxoproline-dependent cleavage of atp, but does not catalyze the decyclization of 5-oxoproline; component b is required for the coupling of atp cleavage to ring-opening of 5-oxoproline to form glutamate (seddon, a. p., li, l., and meister, a. (1984) j. biol. chem. 259, 8091-8094). we describe here the purifications of components a and b to apparent homogeneity and the interactions ...19883360791
bacterial metabolism of side chain fluorinated aromatics: cometabolism of 3-trifluoromethyl(tfm)-benzoate by pseudomonas putida (arvilla) mt-2 and rhodococcus rubropertinctus n657.the tol plasmid-encoded enzymes of the methylbenzoate pathway in pseudomonas putida mt-2 cometabolized 3-trifluoromethyl (tfm)-benzoate. two products, 3-tfm-1,2-dihydroxy-2-hydrobenzoate (3-tfm-dhb) and 2-hydroxy-6-oxo-7,7,7-trifluoro-hepta-2,4-dienoate (7-tfhod) were identified chemically and by spectroscopic properties. tfm-substituted analogues of the metabolites of the methylbenzoate pathway were generally converted at drastically reduced rates. the catechol-2,3-dioxygenase from pseudomonas ...19883365096
bacterial metabolism of side chain fluorinated aromatics: cometabolism of 4-trifluoromethyl(tfm)-benzoate by 4-isopropylbenzoate grown pseudomonas putida jt strains.enzymes of the p-cymene pathway in pseudomonas putida strains cometabolized the intermediate analogue 4-trifluoromethyl(tfm)benzoate. three products, 4-tfm-2,3-dihydro-2,3-dihydroxybenzoate, 4-tfm-2,3-dihydroxybenzoate and 2-hydroxy-6-oxo-7,7,7-trifluorohepta-2,4-dienoate (7-tfhod) were identified chemically and by spectroscopic properties. certain tfm-substituted analogue metabolites of the p-cymene pathway were transformed at drastically reduced rates. hammett type analysis of ring cleavage re ...19883365097
benzylic monooxygenation catalyzed by toluene dioxygenase from pseudomonas putida.toluene dioxygenase, a multicomponent enzyme system known to oxidize mononuclear aromatic hydrocarbons to cis-dihydrodiols, oxidized indene and indan to 1-indenol and 1-indanol, respectively. in addition, the enzyme catalyzed dioxygen addition to the nonaromatic double bond of indene to form cis-1,2-indandiol. the oxygen atoms in 1-indenol and cis-1,2-indandiol were shown to be derived from molecular oxygen, whereas 70% of the oxygen in 1-indanol was derived from water. all of the isolated produ ...19883365392
specific labeling of the essential cysteine residue of l-methionine gamma-lyase with a cofactor analogue, n-(bromoacetyl)pyridoxamine phosphate.l-methionine gamma-lyase from pseudomonas putida is composed of four identical polypeptide chains and contains four cysteinyl residues per subunit. we have found one of them catalytically essential by its specific cyanylation with 2-nitro-5-thiocyanobenzoic acid. we have shown its essentiality also with n-(bromoacetyl)pyridoxamine 5'-phosphate (bapmp), which is a cofactor analogue and also an affinity-labeling agent. the kinetic data show that the apoenzyme forms a binary complex with bapmp prio ...19883365412
cloning and characterization of a gene from rhizobium melilotii 2011 coding for ribosomal protein s1.a 7 kb chromosomal dna fragment from r. melilotii was cloned, which complemented temperature-sensitivity of an e. coli amber mutant in rpsa, the gene for ribosomal protein s1 (es1). from complementation and maxicell analysis a 58 kd protein was identified as the homolog of protein s1 (rs1). dna sequence analysis of the r. melilotii rpsa gene identified a protein of 568 amino acids, which showed 47% identical amino acid homology to protein s1 from e. coli. the rs1 protein lacked the two cys resid ...19883368316
transfer and expression of mesophilic plasmid-mediated degradative capacity in a psychrotrophic bacterium.a psychrotrophic bacterium, originally isolated from a natural aquatic environment, was characterized and identified as pseudomonas putida q5 for use as a representative recipient for biodegradative genes from a mesophilic microorganism. the tol plasmid pwwo of the mesophile p. putida paw1 was successfully transferred by conjugation to the naturally isolated psychrotroph p. putida q5, as shown by plasmid analysis by agarose gel electrophoresis. expression of the genes encoded by the mesophilic t ...19883377489
kinetics and mechanism of benzoylformate decarboxylase using 13c and solvent deuterium isotope effects on benzoylformate and benzoylformate analogues.benzoylformate decarboxylase (benzoylformate carboxy-lyase, bfd; ec 4.1.1.7) from pseudomonas putida is a thiamine pyrophosphate (tpp) dependent enzyme which converts benzoylformate to benzaldehyde and carbon dioxide. the kinetics and mechanism of the benzoylformate decarboxylase reaction were studied by solvent deuterium and 13c kinetic isotope effects with benzoylformate and a series of substituted benzoylformates (pch3o, pch3, pcl, and mf). the reaction was found to have two partially rate-de ...19883378056
difference between amino acid residues in the metal-ligand environments of iron- and manganese-superoxide dismutases.alignment of the amino acid sequences of the pseudomonas ovalis and photobacterium leiognathi iron-superoxide dismutases (fe-sods) with the known sequences of the manganese-superoxide dismutases (mn-sods) shows that both types of sod are highly homologous (33-53% identity) and share residues for the metal coordination. the amino acid residues that form the environment of the metal ions appear to be also conserved between the fe- and mn-sods, except that the phe-84 and gln-154 in the mn-sods are ...19883382418
purification and properties of 4-hydroxyphenylacetic acid 3-hydroxylase from pseudomonas putida.4-hydroxyphenylacetic acid 3-hydroxylase is a key enzyme in the pathway for the microbial degradation of phenylalanine, tyrosine and many aromatic amines. this enzyme was purified to homogeneity from pseudomonas putida by affinity chromatography. the protein had a molecular weight of 91,000 and was a dimer of identical subunits. it was a typical external flavoprotein monooxygenase and showed an absolute requirement of nadh for activity. the enzyme had a ph optimum of 7.5 and the km values for 4- ...19883401220
oxidation of substituted phenols by pseudomonas putida f1 and pseudomonas sp. strain js6.the biodegradation of benzene, toluene, and chlorobenzenes by pseudomonas putida involves the initial conversion of the parent molecules to cis-dihydrodiols by dioxygenase enzyme systems. the cis-dihydrodiols are then converted to the corresponding catechols by dihydrodiol dehydrogenase enzymes. pseudomonas sp. strain js6 uses a similar system for growth on toluene or dichlorobenzenes. we tested the wild-type organisms and a series of mutants for their ability to transform substituted phenols af ...19883415220
degradation of trichloroethylene by toluene dioxygenase in whole-cell studies with pseudomonas putida f1.toluene-induced cells of pseudomonas putida f1 removed trichloroethylene from growth media at a significantly greater initial rate than the methanotroph methylosinus trichosporium ob3b. with toluene-induced p. putida f1, the initial degradation rate varied linearly with trichloroethylene concentration over the range of 8 to 80 microm (1.05 to 10.5 ppm). at 80 microm (10.5 ppm) trichloroethylene and 30 degrees c, the initial rate was 1.8 nmol/min per mg of total cell protein, but the rate decreas ...19883415234
similarity of the e1 subunits of branched-chain-oxoacid dehydrogenase from pseudomonas putida to the corresponding subunits of mammalian branched-chain-oxoacid and pyruvate dehydrogenases.the genes encoding proteins responsible for activity of the e1 component of branched-chain-oxoacid dehydrogenase of pseudomonas putida have been subcloned and the nucleotide sequence of this region determined. open reading frames encoding e1 alpha (bkda1, 1233 bp) and e1 beta (bkda2, 1020 bp) were identified with the aid of the n-terminal sequence of the purified subunits. the mr of e1 alpha was 45,158 and of e1 beta was 37,007, both calculated without n-terminal methionine. the deduced amino ac ...19883416875
construction of chlorobenzene-utilizing recombinants by progenitive manifestation of a rare event.separate continuous cultures of pseudomonas putida r5-3, grown on toluene, and pseudomonas alcaligenes c-o, grown on benzoate, were concentrated and continuously amalgamated on a ceramic bead column, which was subjected to a continuous stream of chlorobenzene vapors. a recombinant strain, p. putida cb1-9, was isolated in less than 1 month. p. alcaligenes c-0 grew on benzoate and 3-chlorobenzoate but not on toluene, p. putida r5-3 grew on benzoate and toluene but not on 3-chlorobenzoate, and neit ...19873426217
laboratory and clinical evaluation of isolation media for campylobacter jejuni.six selective isolation media were evaluated for their ability to support the growth of campylobacter jejuni. colony counts of 70 isolated strains of c. jejuni and recovery studies on these strains in simulated positive feces samples demonstrated that bolton and hutchinson' charcoal, cefoperazone, deoxycholate agar and karmali's charcoal-based selective medium produced the highest recovery rates with the greatest suppression of other fecal flora. c. jejuni colonies were more easily recognized on ...19873429621
a catecholic 9,10-seco steroid as a product of aerobic catabolism of cholic acid by a pseudomonas sp.a mutant of the efficient bile acid-utilizing pseudomonas putida atcc 31752 was found to accumulate three major catabolites on aerobic growth on cholic acid. one of these catabolites was isolated and identified as 3,4,7,12 beta-tetrahydroxy-9,10-seco-1,3,5(10)-androstatriene-9,17-dione (2). this is the first catecholic 9,10-secosteroid isolated from the microbial degradation of bile acids or sterols and confirms the role of such secosteroids in the microbial degradative pathway for steroids.19863445293
benzene dioxygenase in pseudomonas putida. subunit composition and immuno-cross-reactivity with other aromatic dioxygenases.the terminal oxygenase component of benzene dioxygenase from pseudomonas putida strain ml2 was shown to contain two subunits, of mr 54,500 and 23,500, by sds/polyacrylamide-gel electrophoresis. the native mr of the terminal oxygenase was estimated to be 168,000 +/- 4000. polyclonal antibodies raised against each of the subunits cross-reacted with two polypeptides in cell-free extracts from toluene-grown pseudomonas putida strain n.c.i.b. 11767. the mr values of these polypeptides were similar to ...19873446181
bacterial flora in bottled uncarbonated mineral drinking water.a quantitative study of bacterial populations in mineral water was carried out. samples were stored at 6 and 20 degrees c, and the colony counts were determined on tryptone agar plates incubated at 22 and 37 degrees c. samples were collected from the spring source in sterile glass flasks and from the bottling factory in conventional plastic and glass containers. in both cases, the initial population (10(1)-10(2) cfu/ml water) increased to 10(5)-10(6) cfu/ml after 3 days storage as determined fro ...19873446349
regulatory circuits controlling transcription of tol plasmid operon encoding meta-cleavage pathway for degradation of alkylbenzoates by pseudomonas.tol plasmid pwwo of pseudomonas putida contains two operons that specify a pathway for the degradation of aromatic hydrocarbons. the 'upper' operon encodes enzymes for the oxidation of toluene to benzoate and xylenes to toluates, whereas the meta-cleavage operon specifies the further oxidation of benzoate and toluates. transcription of the upper pathway operon is positively regulated by the xylr protein, which is activated by toluene/xylenes and their alcohol catabolic products, in combination w ...19873448461
in vitro activity of imipenem towards gram-negative bacilli.the authors studied the in vitro antimicrobial activity of imipenem towards 355 gram-negative bacterial strains, taking into particular consideration unusual or dangerous species. the study was carried out on a comparative basis with piperacillin, cefotaxime, ceftazidime and gentamicin. ninety percent of the fermentative gram-negative strains were inhibited at concentrations less than or equal to 2 mg/l. imipenem inhibited 100% of strains of alcaligenes faecalis, alcaligenes denitrificans, flavo ...19863466723
the xyls gene positive regulator of tol plasmid pwwo: identification, sequence analysis and overproduction leading to constitutive expression of meta cleavage operon.the pseudomonas putida tol plasmid pwwo carries an operon that specifies a meta-cleavage pathway for the catabolism of benzoate and toluates whose transcription is positively regulated by the xyls gene product. stimulation of transcription of the operon is thought to result from activation of this protein by pathway substrates/effectors. in the present study, overexpression of the xyls gene has led to identification of the regulator as a 33 kda protein. overexpression of xyls also resulted in pa ...19873475526
formaldehyde dismutase, a novel nad-binding oxidoreductase from pseudomonas putida f61.a novel enzyme, formaldehyde dismutase, was purified and crystallized from the cell extract of an isolated bacterium, pseudomonas putida f61. the enzyme catalyzes the dismutation of aldehydes and alcohol:aldehyde oxidoreduction in the absence of an exogenous electron acceptor. the enzyme is composed of four identical subunits with a mr of 44 000. each subunit contains 1 mol nad(h) and 2 mol zinc/mol. the ratio of nad+ and nadh in a crystalline preparation of the enzyme was about 7:3. the enzyme- ...19863514215
leads from the mmwr. reported contamination of heparin sodium with pseudomonas putida. 19863517390
[selection of escherichia coli and pseudomonas putida cultures with an enhanced resistance to immobilization on polyacrylamide gel].clones of escherichia coli (a4, a70, g60) and pseudomonas putida (a70, g30) with an elevated resistance to the process of immobilization in polyacrylamide gel and to the action of monomeric acrylamide were selected from the parent e. coli ibpm b115 and p. putida. the isolated cultures remained resistant to the above actions for a long time. the frequency at which cells with the elevated resistance appeared was comparable with the frequency of bacterial mutations. the plasmid analysis did not rev ...19863517599
the xylabc promoter from the pseudomonas putida tol plasmid is activated by nitrogen regulatory genes in escherichia coli.the xylabc promoter (op1), located on the tol plasmid of pseudomonas putida contains sequences homologous to the conserved regions found in nitrogen fixation (nif) promoters and in other promoters subject to nitrogen control. xyla-lac fusions were constructed in order to monitor expression from the op1 promoter in escherichia coli. transcription was activated in the presence of the heterologous regulatory genes ntrc or nifa from klebsiella pneumoniae as well as by the homologous p. putida regula ...19863520241
identification of legionella pneumophila with commercially available immunofluorescence test. 19863522642
vector for regulated expression of cloned genes in a wide range of gram-negative bacteria.a pkt231-based broad-host-range plasmid vector was constructed which enabled regulation of expression of cloned genes in a wide range of gram-negative bacteria. this vector, pnm185, contained upstream of its ecori, ssti, and sstii cloning sites the positively activated pm twin promoters of the tol plasmid and xyls, the gene of the positive regulator of these promoters. expression of cloned genes was induced with micromolar quantities of benzoate or m-toluate, the inexpensive coinducers of the pm ...19863525513
molecular cloning of genes encoding branched-chain keto acid dehydrogenase of pseudomonas putida.we cloned the structural genes for the individual subunits of the branched-chain keto acid dehydrogenase multienzyme complex on a 7.8-kilobase ecori-ssti restriction fragment of pseudomonas putida chromosomal dna by cloning into the broad-host-range vector pkt230. a direct selection system for growth on valine-isoleucine agar was achieved by complementation of p. putida branched-chain keto acid dehydrogenase mutants. the recombinant plasmid, pss1-1, increased expression of branched-chain keto ac ...19873549697
characterization of a novel tol-like plasmid from pseudomonas putida involved in 1,2,4-trimethylbenzene degradation.a strain of pseudomonas putida (tmb) was found to resemble p. putida mt-2 (paw1) in its ability to degrade 1,2,4-trimethylbenzene, toluene, m-xylene, and p-xylene via oxidation of a methyl substituent and reaction of the meta fission pathway, but a different regulatory model is suggested. the ability of p. putida tmb to degrade these substrates was encoded by plasmid pgb (85 kilobase pairs), which showed considerable differences in size, restriction patterns, and dna sequence from those of plasm ...19873558324
biosynthesis of thiamin. different biosynthetic routes of the thiazole moiety of thiamin in aerobic organisms and anaerobic organisms.the nitrogen atom of glycine was incorporated into the thiazole moiety of thiamin in the aerobic microorganisms bacillus subtilis, pseudomonas putida, saccharomyces cerevisiae, mucor racemosus, neurospora crassa, and emericella nidulans. it was not incorporated in the case of the facultative anaerobic microorganisms escherichia coli and enterobacter aerogenes, which, however, did incorporate the nitrogen atom of tyrosine. these results show that aerobic microorganisms and facultative anaerobic m ...19873566774
[effect of the medium composition on the accumulation of 2-keto-d-gluconic acid in pseudomonas putida cultures].the effect of the composition of the culture medium and the age of the culture on the activities of the enzymes involved in accumulation of 2-ketogluconic acid by pseudomonas putida was studied. the activities of glucose and gluconate dehydrogenases that are responsible for direct oxidation of glucose to 2-ketogluconic acid, were 2-3 times higher during the active growth of the culture than in the stationary phase. the activities of 2-ketogluconokinase and 2-keto-6-phosphogluconate reductase, en ...19873575266
survival of selected bacterial species in sterilized activated carbon filters and biological activated carbon filters.the survival of selected hygienically relevant bacterial species in activated carbon (ac) filters on a bench scale was investigated. the results revealed that after inoculation of the test strains the previously sterilized ac absorbed all bacteria (10(6) to 10(7)). after a period of 6 to 13 days without countable bacteria in the effluent, the numbers of escherichia coli, pseudomonas aeruginosa, and pseudomonas putida increased up to 10(4) to 10(5) cfu/ml of effluent and 10(6) to 10(7) cfu/g of a ...19873579281
recruitment of naphthalene dissimilatory enzymes for the oxidation of 1,4-dichloronaphthalene to 3,6-dichlorosalicylate, a precursor for the herbicide dicamba.pseudomonas putida expresses plasmid-encoded enzymes and regulatory proteins for the dissimilation of naphthalene through salicylate and the alpha-keto acid pathway. a strain of p. putida (nah:tn5/g67) defective in salicylate hydroxylase (nahg) was assessed for its ability to oxidize 1,4-dichloronaphthalene. washed cell suspensions were shown to accumulate 3,6-dichlorosalicylate, which, after further chemical treatment, yields the herbicide dicamba (3,6-dichloro-2-methoxybenzoate). however, the ...19873584076
expression of degradative genes of pseudomonas putida in caulobacter crescentus.the recombinant plasmid rp4-tol was transferred into caulobacter crescentus at a high frequency, and the plasmid was maintained for at least 50 generations. c. crescentus cells which contained rp4-tol grew on all the aromatic compounds that the plasmid normally allowed pseudomonas putida to grow on. reciprocal transfers from c. crescentus donor to p. putida or escherichia coli recipients were less efficient and occurred at frequencies of approximately 10(-3). some representative tol-specified en ...19873597317
comparison of a gelation and a chromogenic limulus (lal) assay for the detection of gram-negative bacteria, and the application of the latter assay to milk.when a chromogenic limulus amoebocyte lysate (lal) assay and a tube gelation lal assay were compared for the detection of gram-negative bacteria using a strain of pseudomonas putida, the detection level (approximately 10(3) cfu/ml) and cost of the assays were approximately the same for both assays but the reading was more precise for the chromogenic substrate assay. a modified chromogenic assay was devised for detection of ps. putida in milk.19873597923
pseudomonas putida. newly recognized pathogen in patients with cancer.pseudomonas putida was recovered from blood culture specimens between 1980 and 1985 in 15 patients with cancer. no isolates were found in specimens obtained before 1980. eight patients were considered to have septicemia (more than one positive blood culture result plus clinical signs of infection). septicemia was monomicrobial in three of those eight patients and polymicrobial in five. of these eight patients, one had pneumonia and three had phlebitis, cellulitis, or both at the site of the veno ...19873605136
cloning and complete nucleotide sequence determination of the catb gene encoding cis,cis-muconate lactonizing enzyme.the enzyme, cis,cis-muconate lactonizing enzyme i (mlei; ec 5.5.1.1), has been proposed to play a key role in the beta-ketoadipate pathway of benzoate degradation. a 10.2-kb ecori fragment isolated from a pseudomonas putida genomic library complemented a mutant deficient in this enzyme. the mlei coding gene, catb, was localized to a 1.6-kb fragment which was sequenced by the dideoxy chain termination method. mlei was purified 25-fold from crude extracts of benzoate-grown p. putida prs2015 harbor ...19873609743
overproduction of the xyls gene product and activation of the xyldlegf operon on the tol plasmid.the effect of high-level expression of the regulatory gene xyls of the pseudomonas putida tol plasmid on the activation of the xyldlegf operon was investigated in escherichia coli. the xyls gene was placed downstream from the tac promoter, and the resultant fusion was cloned in cis to the xyldlegf operon. the expression of the operon was monitored by the level of catechol 2,3-dioxygenase, whose structural gene xyle was placed directly after the operator-promoter region of xyldlegf. xyls transcri ...19873611023
18o studies on the 5-oxoprolinase reaction. evidence for a phosphorylated tetrahedral intermediate.5-oxoprolinase catalyzes a reaction in which the cleavage of atp to adp and pi and the decyclization of 5-oxoproline to form glutamate are coupled. when the reaction catalyzed by 5-oxoprolinase of pseudomonas putida was carried out to 90% completion in h2(18)o, the residual 5-oxoproline was found to contain 18o in the amide carbonyl oxygen atom. such isotopic incorporation was not observed in similar studies with a subunit of the enzyme which catalyzes 5-oxoproline-dependent atpase and formation ...19873611103
nucleotide sequence and expression of gene nahh of plasmid nah7 and homology with gene xyle of tol pwwo.the enzyme catechol 2,3-dioxygenase (c23o) encoded by the nahh gene of plasmid nah7 converts catechol to alpha-hydroxymuconic epsilon-semialdehyde in pseudomonas putida. we have cloned this structural gene into vectors puc18 and pkt240, determined the nucleotide sequence and deduced the amino acid sequence. in comparison to the gene xyle of the tol plasmid pwwo which encodes a similar c230 enzyme [nakai et al. j. biol. chem. (1983b), 2923-2928], the respective g + c contents were 55% and 57%, th ...19873623105
use of salicylate to estimate the "threshold" inducer level for de novo synthesis of the phenol-degrading enzymes in pseudomonas putida strain h.a special approach was used to elucidate the "threshold" inducer concentration for coordinative de novo synthesis of phenol hydroxylase(s), catechol 2,3-dioxygenase and the 2-hydroxymuconic semialdehyde-metabolizing enzymes which initiate phenol catabolism in pseudomonas putida strain h. it is based on cell-precultivation with glucose (as the carbon and energy source) in the presence of different concentrations of sodium salicylate which proved to be a potent non-metabolizable inducer in strain ...19873656095
amino acid sequence of iron-superoxide dismutase from pseudomonas ovalis.the amino acid sequence of iron-superoxide dismutase from pseudomonas ovalis was deduced by the analyses of peptides derived from limited hydrolysis of the aminoethylated or pyridylethylated apoprotein with trypsin, staphylococcus aureus v8 protease, and dilute acid hydrolysis. the polypeptide chain contains 195 amino acid residues and has a calculated mr of 21,421. the sequence is highly homologous (65% identity) to the recently published sequence of the iron-superoxide dismutase from photobact ...19873666146
nucleotide sequencing and characterization of the genes encoding benzene oxidation enzymes of pseudomonas putida.the nucleotide sequence of the genes from pseudomonas putida encoding oxidation of benzene to catechol was determined. five open reading frames were found in the sequence. four corresponding protein molecules were detected by a dna-directed in vitro translation system. escherichia coli cells containing the fragment with the four open reading frames transformed benzene to cis-benzene glycol, which is an intermediate of the oxidation of benzene to catechol. the relation between the product of each ...19873667527
the effect of lipophilic weak acids on the segregational stability of tol plasmids in pseudomonas putida.the effect of various lipophilic weak acids on the stability of certain tol plasmids was investigated. benzoate induced deletion of tol plasmid dna in pseudomonas putida mt15, followed by loss of the plasmid; this effect was ph- and concentration-dependent, suggesting that undissociated benzoic acid was a more effective curing agent than the benzoate anion. plasmid loss always approached a frequency of 100% after a lag and apparently depended on the prior occurrence of deletions, although delete ...19873668501
[in vitro bacteriostatic activity of piperacillin sodium against pseudomonas].one hundred and twenty-two strains of pseudomonas isolated from diverse pathological processes were typified. in vitro activity of piperacillin (antibiogram and mic) was studied and compared with another two semisynthetic penicillins, carbenicillin and mezlocillin, and two aminoglycosides, amikacin and gentamicin. the strains isolated corresponded to: pseudomonas aeruginosa (116), pseudomonas cepacia (3), pseudomonas fluorescens (1) and pseudomonas putida (1). it was found that 110 pseudomonas a ...19863685380
the amino acid sequence of a delta 5-3-oxosteroid isomerase from pseudomonas putida biotype b.we have determined the primary structure of a delta 5-3-oxosteroid isomerase from pseudomonas putida biotype b. the enzyme is a dimeric protein of two identical subunits, each consisting of a polypeptide chain of 131 residues and a mr = 14,536. the intact s-carboxymethyl protein was sequenced from the nh2 terminus using standard automated edman degradation and automated edman degradation using fluorescamine treatment at known prolines to suppress background. the isomerase was fragmented using cn ...19863700400
l-methionine gamma-lyase from pseudomonas putida and aeromonas. 19873657560
high-resolution crystal structure of cytochrome p450cam.the crystal structure of pseudomonas putida cytochrome p450cam with its substrate, camphor, bound has been refined to r = 0.19 at a normal resolution of 1.63 a. while the 1.63 a model confirms our initial analysis based on the 2.6 a model, the higher resolution structure has revealed important new details. these include a more precise assignment of sequence to secondary structure, the identification of three cis-proline residues, and a more detailed picture of substrate-protein interactions. in ...19873656428
purification and characterization of a novel enzyme, n-carbamoylsarcosine amidohydrolase, from pseudomonas putida 77.n-carbamoylsarcosine amidohydrolase, a novel enzyme involved in the microbial degradation of creatinine in pseudomonas putida 77, was purified 27-fold to homogeneity with a 63% overall recovery through simple purification procedures including successive ammonium sulfate fractionation, deae-cellulose chromatography, and crystallization. the relative molecular mass of the native enzyme estimated by the ultracentrifugal equilibrium method is 102,000 +/- 5000, and the subunit mr is 27,000. the km an ...19863745168
influence of para-substituents on the oxidative metabolism of o-nitrophenols by pseudomonas putida b2.pseudomonas putida b2 is able to grow on o-nitrophenol (onp) as the sole source of carbon and nitrogen. onp was converted by a nitrophenol oxygenase to nitrite and catechol. catechol was then attacked by a catechol 1,2-dioxygenase and further degraded through an ortho-cleavage pathway. onp derivatives which were para-substituted with a methyl-, chloro-, carboxy-, formyl- or nitro-group failed to support growth of strain b2. relevant catabolic enzymes were characterized to analyze why these deriv ...19863752997
primary alkylsulphatase activities of the detergent-degrading bacterium pseudomonas c12b. purification and properties of the p1 enzyme.the p1 primary alkylsulphatase of pseudomonas c12b was purified 1500-fold to homogeneity by a combination of streptomycin sulphate precipitation of nucleic acids, (nh4)2so4 fractionation and chromatography on columns of deae-cellulose, sephacryl s-300 and butyl-agarose. the protein was tetrameric with an mr of 181000-193000, and exhibited maximum activity at ph 6.1. primary alkyl sulphates of carbon-chain length c1-c5 or above c14 were not substrates, but the intermediate homologues were shown t ...19863753455
identification of cis-diols as intermediates in the oxidation of aromatic acids by a strain of pseudomonas putida that contains a tol plasmid.pseudomonas putida bg1 was isolated from soil by enrichment with p-toluate and selection for growth with p-xylene. other hydrocarbons that served as growth substrates were toluene, m-xylene, 3-ethyltoluene, and 1,2,4-trimethylbenzene. the enzymes responsible for growth on these substrates are encoded by a large plasmid with properties similar to those of tol plasmids isolated from other strains of pseudomonas. treatment of p. putida bg1 with nitrosoguanidine led to the isolation of a mutant stra ...19863711022
crystal structure of substrate-free pseudomonas putida cytochrome p-450.the crystal structure of pseudomonas putida cytochrome p-450cam in the substrate-free form has been refined at 2.20-a resolution and compared to the substrate-bound form of the enzyme. in the absence of the substrate camphor, the p-450cam heme iron atom is hexacoordinate with the sulfur atom of cys-357 providing one axial heme ligand and a water molecule or hydroxide ion providing the other axial ligand. a network of hydrogen-bonded solvent molecules occupies the substrate pocket in addition to ...19863768350
comparative analysis of different pseudomonas strains that degrade cinnamic acid.strains of pseudomonas stutzeri (cinns) and pseudomonas putida (cinnp and cinnw) isolated from soil with cinnamic acid as the sole carbon source were found to be simultaneously adapted to grow on phenylpropionic and p-hydroxybenzoic acids. in cinnamic acid-grown cultures, phenylpropionic acid was isolated. a catabolic plasmid of approximately equal to 75 kilobase pairs encoding the metabolism of cinnamic acid was found in strains cinnp and cinns.19863777934
chromosomal location of tol plasmid dna in pseudomonas putida.the soil isolate pseudomonas putida mw1000 can grow on toluene and other hydrocarbons; in this respect it is similar to strains of pseudomonas which carry the tol plasmid. by conjugation experiments, the genes conferring these growth abilities have been shown to be located on the bacterial chromosome, linked to vil and catb. a 56-kilobase segment of the bacterial chromosome of mw strains carrying the tol genes can transpose to the incp-1 plasmid r18-18. physical analysis of these tol r18-18 hybr ...19863782038
cadmium-binding proteins in pseudomonas putida: pseudothioneins.pseudomonas putida adapted to growth in 3 mm cadmium. the resistance mechanism involved complexation of cadmium in polyphosphate granules, changes in the structure of the cell membrane and induction of three cysteine-rich, low molecular weight proteins (3500-7000) containing 4 to 7 g-atoms per mole of cadmium, zinc, and copper. each protein was produced during a different phase of growth, and the smallest protein (3500) was released into the environment when the cells lysed at the end of the exp ...19863709466
discovery of a cutinase-producing pseudomonas sp. cohabiting with an apparently nitrogen-fixing corynebacterium sp. in the phyllosphere.a phyllospheric bacterial culture, previously reported to partially replace nitrogen fertilizer (b. r. patti and a. k. chandra, plant soil 61:419-427, 1981) was found to contain a fluorescent pseudomonas which was identified as pseudomonas putida and a corynebacterium sp. the p. putida isolate was found to produce an extracellular cutinase when grown in a medium containing cutin, the polyester structural component of plant cuticle. the corynebacterium sp. grew on nitrogen-free medium but could n ...19873793714
adaptation of pseudomonas putida mt-2 to growth on aromatic amines.pseudomonas putida mt-2 (atcc 33015) carrying the tol plasmid pww0 could adapt to growth on the aromatic amines aniline and m- and p-toluidine. in strain ucc2, a derivative adapted to rapid growth on these compounds, they were oxidatively deaminated to catechol or 4-methylcatechol, which in turn were dissimilated by a meta-cleavage pathway. the aniline/toluidine oxygenase and the meta-cleavage pathway enzymes were inducible by aromatic amines. evidence is presented that in strain ucc2, plasmid p ...19863794647
[catabolic pathway of biphenyl controlled by the plasmid pbs241 in pseudomonas putida bs893]. 19863720511
amino acid and sequence analysis of the cytochrome and flavoprotein subunits of p-cresol methylhydroxylase.the flavocytochrome p-cresol methylhydroxylase from pseudomonas putida has been reported to have a mr of 114,000 and to consist of two subunits, a flavoprotein and a cytochrome c, each with a mr of 58,000. recent x-ray crystallographic data from our laboratories [shamala, n., lim, l. w., mathews, f. s., mcintire, w., singer, t. p., & hopper, d. j. (1986) proc. natl. acad. sci. u.s.a. 83, 4626-4630], however, indicate an alpha 2 beta 2 structure and a much lower molecular mass (approximately 8000 ...19863790500
amidohydrolysis of n-methylhydantoin coupled with atp hydrolysis.a new enzyme, n-methylhydantoin amidohydrolase, was highly purified from pseudomonas putida 77: it catalyzes the hydrolysis of n-methylhydantoin to n-carbamoylsarcosine with the concomitant stoichiometric cleavage of atp to adp and orthophosphate. the enzyme absolutely requires atp, mg2+ and k+ for the n-methylhydantoin hydrolysis. the rapid and complete degradation of n-methylhydantoin during the cultivation of p. putida 77, which rapidly degrades creatinine via only n-methylhydantoin and which ...19873827889
cloning of pseudomonas sp. strain cbs3 genes specifying dehalogenation of 4-chlorobenzoate.the degradation of 4-chlorobenzoate (4-cba) by pseudomonas sp. strain cbs3 is thought to proceed first by the dehalogenation of 4-cba to 4-hydroxybenzoate (4-hba), which is then metabolized following the protocatechuate branch of the beta-ketoadipate pathway. the cloning of the 4-cba dehalogenation system was carried out by constructing a gene bank of pseudomonas sp. strain cbs3 in pseudomonas putida. hybrid plasmid ppsa843 contains a 9.5-kilobase-pair fragment derived from the chromosome of pse ...19863759912
identification of the promoter of the pseudomonas gene coding for carboxypeptidase g2.a 213-bp region of noncoding dna upstream of the atg start codon of the pseudomonas carboxypeptidase g2 (cpg2) structural gene has been shown to contain the cpg2 promoter. the mrna start point (+1) on the dna sequence has been identified by mapping the 5' end of the cpg2 transcript. the identified promoter region contains a -10 region (tataag) that closely resembles the escherichia coli consensus sequence (tataat), but has no easily recognisable -35 region. the lack of homology in the -35 region ...19853839252
genes specifying degradation of 3-chlorobenzoic acid in plasmids pac27 and pjp4.all of the structural genes for 3-chlorobenzoate degradation are clustered in a 4.2-kilobase (kb) region of plasmid pac25 (or pac27) in pseudomonas putida. an approximate 10-kb dna segment containing three structural genes for chlorocatechol metabolism present on plasmid pjp4 in alcaligenes eutrophus shows homology with the above 4.2-kb region of pac27. in spite of the detectable sequence homology in the structural genes present on both plasmids, the regulation of their expression seems quite di ...19853856842
hydrodynamic characterization of the size and shape of atropinesterase from pseudomonas putida.atropinesterase from pseudomonas putida has been investigated by means of different ultracentrifugation methods under native and denaturing conditions. the following quantities were determined: sedimentation coefficient, translational diffusion and friction coefficient, partial specific volume and molecular weight. from these data the size, shape and hydration of the enzyme molecule in solution were estimated. the results suggest that atropinesterase is a globular protein which consists of a sin ...19873828356
cyclocoelum mutabile infection and aortic rupture in an american coot (fulica americana).an american coot (fulica americana) was found dead within the enclosed research compound of the south central poultry research laboratory at mississippi state, mississippi. gross and microscopic examinations revealed the bird to be in good body condition; however, blood from the beak cavity and external nares was present. biliary congestion, hemopericardium, blood-filled air sacs, and a ruptured, ascending aorta were also noted. nineteen trematodes (cyclocoelum mutabile) were found within the bo ...19853885932
transcriptional control of the nah and sal hydrocarbon-degradation operons by the nahr gene product.the positively regulated nah and sal operons of the nah7 plasmid from pseudomonas putida encode the enzymes for metabolism of naphthalene via salicylate. to study their coordinate regulation, a 6-kb dna fragment containing the entire naha gene (encoding naphthalene dioxygenase), the gene of the nah operon, was cloned into a rsf1010 plasmid derivative. analysis of expression of naha from the nah promoter in either escherichia coli or pseudomonas putida showed that a 1.6-kb dna fragment from the n ...19853908220
dienelactone hydrolase from pseudomonas sp. strain b13.dienelactone hydrolase (ec 3.1.1.45) catalyzes the conversion of cis- or trans-4-carboxymethylenebut-2-en-4-olide (dienelactone) to maleylacetate. an approximately 24-fold purification from extracts of 3-chlorobenzoate-grown pseudomonas sp. strain b13 yielded a homogeneous preparation of the enzyme. the purified enzyme crystallized readily and proved to be a monomer with a molecular weight of about 30,000. each dienelactone hydrolase molecule contains two cysteinyl side chains. one of these was ...19873804973
use of cloned genes of pseudomonas tol plasmid to effect biotransformation of benzoates to cis-dihydrodiols and catechols by escherichia coli cells.dna fragments containing the xyld and xyll genes, which specify the broad-specificity enzymes toluate-1,2-dioxygenase and 3,5-cyclohexadiene-1,2-diol-1-carboxylic acid dehydrogenase, respectively, of tol plasmid pww0-161 of pseudomonas putida have previously been cloned in the pbr322 vector plasmid (p.r. lehrbach, j. zeyer, w. reinecke, h.-j. knackmuss, and k. n. timmis, j. bacteriol. 158:1025-1032, 1984). in this study, escherichia coli cells containing hybrid plasmids carrying the cloned xyld ...19853911905
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