Publications
| Title | Abstract | Year Filter | PMID(sorted ascending) Filter |
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| a t7 rna polymerase-based system for the construction of pseudomonas strains with phenotypes dependent on tol-meta pathway effectors. | a general method to construct recombinant pseudomonas putida (and related bacteria), which transcribe specific genes inserted into their chromosome in response to the presence of alkyl- and halobenzoates, has been developed. the system is based on the ability of the t7 rna polymerase (t7pol) to initiate transcription from cognate promoter sequences located upstream from cloned genes. a specialized transposon, mini-tn5 xyls/pm::t7pol, was constructed which contains the structural t7 gene 1 downst ... | 1993 | 8244019 |
| the specific growth rate of pseudomonas putida paw1 influences the conjugal transfer rate of the tol plasmid. | the kinetics of the conjugal transfer of a tol plasmid were investigated by using pseudomonas putida paw1 as the donor strain and p. aeruginosa pao 1162 as the recipient strain. short-term batch mating experiments were performed in a nonselective medium, while the evolution of the different cell types was determined by selective plating techniques. the experimental data were analyzed by using a mass action model that describes plasmid transfer kinetics. this method allowed analysis of the mating ... | 1993 | 8250565 |
| adaptation of pseudomonas putida s12 to high concentrations of styrene and other organic solvents. | pseudomonas putida s12 could adapt to grow on styrene in a two-phase styrene-water system. acetate was toxic for p. putida s12, but cells were similarly able to adapt to higher acetate concentrations. only by using these acetate-adapted cells was growth observed in the presence of supersaturating concentrations of toxic nonmetabolizable solvents such as toluene. | 1993 | 8250572 |
| quinoline oxidoreductase from pseudomonas putida 86: an improved purification procedure and electron paramagnetic resonance spectroscopy. | quinoline oxidoreductase, an iron-sulfur molybdenum flavoprotein containing flavin adenine dinucleotide and molybdopterin cytosine dinucleotide, was purified from pseudomonas putida 86 to homogeneity. the various electron-transfer centers of the purified enzyme were examined by electron paramagnetic resonance spectroscopy. quinoline deuterated at position 2 was prepared by deuterodecarboxylation of 2-quinolinecarboxylic acid. quinoline added to the enzyme elicited the mo(v) "rapid" type q signal ... | 1993 | 8251516 |
| purification and characterization of pseudomonas putida histidine ammonia-lyase expressed in escherichia coli. | histidine ammonia-lyase (hal) from pseudomonas putida prs1 contains a catalytically important electrophilic center reported to be dehydroalanine. little is known about the origin of this group or its linkage to the protein. to initiate structural studies on this enzyme, p. putida hal was purified from an escherichia coli high-expression clone in which the hal gene (huth) was under the control of the lambda pl promoter on a plasmid vector. in this clone from 6 to 10% of the soluble cell protein a ... | 1993 | 8251759 |
| stabilization of creatinase from pseudomonas putida by random mutagenesis. | creatinase (creatine amidinohydrolase, ec 3.5.3.3) from pseudomonas putida is a homodimer of 45 kda subunit molecular mass, the three-dimensional structure of which is known at 1.9 a resolution. three point mutants, a109v, v355m, and v182i, as well as one double mutant combining a109v and v355m, and the triple mutant with all three replacements, were compared with wild-type creatinase regarding their physical and enzymological properties. high-resolution crystal data for wild-type creatinase and ... | 1993 | 8251936 |
| evidence for autoregulation of camr, which encodes a repressor for the cytochrome p-450cam hydroxylase operon on the pseudomonas putida cam plasmid. | the regulatory gene camr on the cam plasmid of pseudomonas putida (atcc 17453) negatively controls expression of the cytochrome p-450cam hydroxylase operon (camdcab) for the camphor degradation pathway and is oriented in a direction opposite to that of the camdcab operon. in this study, we examined expression of the camr gene by monitoring the beta-galactosidase activity of camr-lacz translational fusions in p. putida camr and camr+ strains. we found that the camr gene was autogenously regulated ... | 1993 | 8253671 |
| regulation of the catechol 1,2-dioxygenase- and phenol monooxygenase-encoding pheba operon in pseudomonas putida paw85. | in pseudomonas putida paw85, the ortho-cleavage pathway is used for catechol degradation. the 11.4-kb xhoi fragment cloned from phenol degradation plasmid pest1226 into pkt240 (recombinant plasmid pat1140) contains the inducible pheba operon that encodes catechol 1,2-dioxygenase (gene pheb) and phenol monooxygenase (gene phea), the first two enzymes for the phenol degradation pathway. the promoter of the pheba operon is mapped 1.5 kb upstream of the pheb gene. the plasmid pat1140, when introduce ... | 1993 | 8253692 |
| new derivatives of tol plasmid pww0. | two new segregants, ppw1-1 and ppw161-1, of pseudomonas putida were isolated from the stock cultures paw85(pww0) and paw85(pww0-161). strain ppw1-1 had lost its ability to grow on m-xylene but was able to grow on m-toluate. a deletion of the left-hand of transposon tn4651, including the upper-operon genes, had taken place in plasmid pww0mut1, isolated from strain ppw1-1. additional deletions were observed in pww0mut1 after 'benzoate-curing' (plasmids pww0mut15, pww0mut19, pww0mut27). the genes o ... | 1993 | 8254307 |
| [microbial enzymes and their inhibitors]. | several proteolytic enzymes and dehydrogenases of microbial origin were studied with special regard to structure-activity relationship. enzyme genes of zn-proteases, subtilisin and pyroglutamyl aminopeptidase from genus bacillus, prolyl endopeptidases from flavobacterium and aeromonas, and of protease ii from e. coli were cloned, sequenced and overproduced in e. coli, their active site structures being elucidated by chemical modification as well as by site-directed mutagenesis. homology analysis ... | 1993 | 8254479 |
| adaptive mutation: the uses of adversity. | when populations of microorganisms are subjected to certain nonlethal selections, useful mutants arise among the nongrowing cells whereas useless mutants do not. this phenomenon, known as adaptive, directed, or selection-induced mutation, challenges the long-held belief that mutations only arise at random and without regard for utility. in recent years a growing number of studies have examined adaptive mutation in both bacteria and yeast. although conflicts and controversies remain, the weight o ... | 1993 | 8257106 |
| a dna module encoding bph genes for the degradation of polychlorinated biphenyls (pcbs). | in this report we describe the development and construction of a dna module which encodes bph genes for the metabolism of pcbs and which is capable of stable integration into the chromosome of gram negative bacteria. introduction of the bph-module into pseudomonas putida kt2442, pseudomonas sp. strain b13 and its genetically engineered derivative b13fr1 expanded the biodegradative ability of these strains to include biphenyl and 4-chlorobiphenyl. the bph operon was stably inherited under laborat ... | 1993 | 8262365 |
| the effects of modified atmospheres on the growth of psychrotrophic pseudomonads on a surface in a model system. | atmospheres containing concentrations of co2 as low as 20% (balance nitrogen) inhibited the growth of pseudomonas fluorescens and pseudomonas putida on the surface of buffered brain heart infusion agar plates, ph 6.8, incubated at 5 or 15 degrees c in flexible packages. the modified atmospheres decreased the growth rates and reduced the populations attained at the end of the exponential phase of growth, but had no substantial effect on the lag phase. p. fluorescens was less tolerant of co2 than ... | 1993 | 8268059 |
| oxidation of carbazole to 3-hydroxycarbazole by naphthalene 1,2-dioxygenase and biphenyl 2,3-dioxygenase. | naphthalene 1,2-dioxygenase from pseudomonas sp. ncib 9816-4 and biphenyl dioxygenase from beijerinckia sp. b8/36 oxidized the aromatic n-heterocycle carbazole to 3-hydroxycarbazole. toluene dioxygenase from pseudomonas putida f39/d did not oxidize carbazole. transformations were carried out by mutant strains which oxidize naphthalene and biphenyl to cis-dihydrodiols, and with a recombinant e. coli strain expressing the structural genes of naphthalene 1,2-dioxgenase from pseudomonas sp. ncib 981 ... | 1993 | 8270195 |
| growth and plasmid-encoded naphthalene catabolism of pseudomonas putida in batch culture. | the growth characteristics of pseudomonas putida plasmid-harbouring strains which catabolize naphthalene via various pathways in batch culture with naphthalene as the sole source of carbon and energy have been investigated. the strains under study were constructed using the host strain p. putida bs394 which contained various naphthalene degradation plasmids. the highest specific growth rate was ensured by the plasmids that control naphthalene catabolism through the meta-pathway of catechol oxida ... | 1993 | 8270196 |
| mechanism of p-hydroxyphenylacetate-3-hydroxylase. a two-protein enzyme. | p-hydroxyphenylacetate-3-hydroxylase purified from pseudomonas putida is a two-protein enzyme requiring a flavoprotein and a coupling protein for productive hydroxylation (arunachalam, u., massey, v., and vaidyanathan, c. s. (1992) j. biol. chem. 267, 25848-25855). this paper presents information on the mechanism of the enzyme from absorbance and fluorescence stopped-flow studies. the reduction of the substrate-free flavoprotein by nadh was slow and was not altered by the presence of the couplin ... | 1994 | 8276789 |
| responses to nutrient starvation in pseudomonas putida kt2442: analysis of general cross-protection, cell shape, and macromolecular content. | the physiology of pseudomonas putida kt2442 with respect to growth and carbon starvation was studied. during the transition from growth to nongrowth, the cell shape changes from cylindrical to spheric, a change which is accompanied by reductions in cell size, dna and ribosome content, and the rate of total protein synthesis. in addition, a pattern of general cross-protection develops, which enables the cells to survive environmental stresses such as high and low temperatures, elevated osmolarity ... | 1994 | 8282712 |
| a substrate-dependent biological containment system for pseudomonas putida based on the escherichia coli gef gene. | a model substrate-dependent suicide system to biologically contain pseudomonas putida kt2440 is reported. the system consists of two elements. one element carries a fusion between a synthetic lac promoter (pa1-04/03) and the gef gene, which encodes a killing function. this element is contained within a transposaseless mini-tn5 transposon so that it can be integrated at random locations on the pseudomonas chromosome. the second element, harbored by plasmid pcc102, is designed to control the first ... | 1993 | 8285679 |
| use of 5-cyano-2,3-ditolyl tetrazolium chloride for quantifying planktonic and sessile respiring bacteria in drinking water. | direct microscopic quantification of respiring (i.e., viable) bacteria was performed for drinking water samples and biofilms grown on different opaque substrata. water samples or biofilms developed in flowing drinking water were incubated with the vital redox dye 5-cyano-2,3-ditolyl tetrazolium chloride (ctc) and r2a medium. one hour of incubation with 0.5 mm ctc was sufficient to obtain intracellular reduction of ctc to the insoluble fluorescent formazan (ctf) product, which was indicative of c ... | 1993 | 8285688 |
| the preparation of endotoxin-free l-methionine-alpha-deamino-gamma-mercaptomethane-lyase (l-methioninase) from pseudomonas putida. | many types of human and animal tumors have an absolute requirement for methionine. this requirement can be satisfied by homocysteine only in normal cells and tissues. therefore, methionine may be an important target in cancer therapy. to attack this target we have purified endotoxin-free methioninase from pseudomonas putida by a novel and simple procedure. this procedure involves (1) a heat step of the cell extract at 60 degrees c for 8 min, (2) deae-toyopearl ion-exchange chromatography, (3) de ... | 1993 | 8286949 |
| fpta, the fe(iii)-pyochelin receptor of pseudomonas aeruginosa: a phenolate siderophore receptor homologous to hydroxamate siderophore receptors. | the pseudomonas aeruginosa siderophore pyochelin is structurally unique among siderophores and possesses neither hydroxamate- nor catecholate-chelating groups. the structural gene encoding the 75-kda outer membrane fe(iii)-pyochelin receptor fpta has been isolated by plasmid rescue techniques and sequenced. the n-terminal amino acid sequence of the isolated fpta protein corresponded to that deduced from the nucleotide sequence of the fpta structural gene. the mature fpta protein has 682 amino ac ... | 1994 | 8288523 |
| [the role of pyrimidines in the biosynthesis of the fluorescing pigment pyoverdin pm in pseudomonas putida m]. | dihydroorotate was shown to be a predecessor of deoxyquinoline nucleus of a fluorescing enzyme pyoverdin pm in pseudomonas putida m. the data was obtained in experiments with a set of pyr- mutants with different steps of pyrimidine synthesis blocked. a scheme for deoxyquinoline nucleus of the enzyme including dihydroorotate participation is proposed. | 1993 | 8289842 |
| microbial metabolism of quinoline and related compounds. xx. quinaldic acid 4-oxidoreductase from pseudomonas sp. ak-2 compared to other procaryotic molybdenum-containing hydroxylases. | quinaldic acid 4-oxidoreductase from pseudomonas sp. ak-2 catalyses the hydroxylation of quinoline 2-carboxylic (quinaldic acid) to 4-hydroxyquinoline 2-carboxylic acid (kynurenic acid) with concomitant reduction of a suitable electron acceptor. an analogous hydroxylation in para-position relative to the n-heteroatom was only recently described for quinaldine 4-oxidoreductase (de beyer & lingens, 1993, biol. chem. hoppe-seyler 374, 101-110) and for quinaldic acid 4-oxidoreductase from serratia m ... | 1993 | 8292263 |
| the role of lysine 166 in the mechanism of mandelate racemase from pseudomonas putida: mechanistic and crystallographic evidence for stereospecific alkylation by (r)-alpha-phenylglycidate. | the mechanism of irreversible inactivation of mandelate racemase (mr) from pseudomonas putida by alpha-phenylglycidate (alpha pga) has been investigated stereochemically and crystallographically. the (r) and (s) enantiomers of alpha pga were synthesized in high enantiomeric excess (81% ee and 83% ee, respectively) using sharpless epoxidation chemistry. (r)-alpha pga was determined to be a stereospecific and stoichiometric irreversible inactivator of mr. (s)-alpha pga does not inactivate mr and a ... | 1994 | 8292591 |
| a mutagenesis system utilizing a tn1722 derivative containing an escherichia coli-specific vector plasmid: application to pseudomonas species. | a novel transposon (tn) mutagenesis system for gram- non-enteric bacteria was developed which allowed rapid and one-step cloning of the mutated region in escherichia coli. the tn constructed was tn1722-299km, a tn1722 derivative containing a kmr gene and the entire sequence of an e. coli-specific plasmid, pacyc184. the hybrid plasmid consisting of tn1722-299km and the transfer genes of plasmid r388 was conjugally transferred from e. coli to pseudomonas putida or p. aeruginosa, and selection of t ... | 1993 | 8294012 |
| construction of chromosomal reca mutants of pseudomonas putida ppg2. | the reca gene of pseudomonas putida ppg2 was cloned by complementation of the reca mutations of escherichia coli strains dh5 alpha and hb101. the nucleotide sequence of the dna fragment was determined and shown to contain reca and a downstream partial open reading frame. two mutants of p. putida ppg2, strains js387 and js388, were constructed by insertional inactivation of reca with a tetracycline-resistance gene in both orientations. both mutants acquired sensitivity to methyl methanesulfonate ... | 1993 | 8294013 |
| putative functions of phenylalanine-350 of pseudomonas putida cytochrome p-450cam. | cytochrome p-450cam hydroxylates d-camphor, using molecular oxygen and reducing equivalents transferred via putidaredoxin. we constructed mutant genes in which phe-350 of p-450cam was replaced by leu, tyr, or his by site-directed mutagenesis, expressed them in escherichia coli, purified the mutant proteins, and compared their enzymic properties with those of the wild type p-450cam. nadh oxidation rate of the tyr mutant in the reconstituted system with putidaredoxin and putidaredoxin reductase wa ... | 1994 | 8305479 |
| analysis of the dna damage-mediated induction of pseudomonas putida and pseudomonas aeruginosa lexa genes. | a fusion between the lexa gene of pseudomonas aeruginosa and pseudomonas putida and the lacz gene was constructed in vitro and cloned in a mini-tn5 transposon derivative to obtain chromosomal insertions which enable to quantitatively examine their transcriptional regulation in both pseudomonas and e. coli. analysis of dna damage-mediated induction of these lexa-lacz fusions showed that expression of p. putida and p. aeruginosa lexa genes was always higher and earlier than the expression of the l ... | 1993 | 8319897 |
| the bkdr gene of pseudomonas putida is required for expression of the bkd operon and encodes a protein related to lrp of escherichia coli. | branched-chain keto acid dehydrogenase is a multienzyme complex which is required for the metabolism of the branched-chain amino acids in pseudomonas putida. the structural genes encoding all four proteins of the bkd operon have been cloned, and their nucleotide sequences have been determined (g. burns, k. t. madhusudhan, k. hatter, and j. r. sokatch, p. 177-184 in s. silver, a. m. chakrabarty, b. iglewski, and s. kaplan [ed.], pseudomonas: biotransformations, pathogenesis, and evolving biotechn ... | 1993 | 8320210 |
| a new amino acid racemase with threonine alpha-epimerase activity from pseudomonas putida: purification and characterization. | we have found that pseudomonas putida atcc 17642 cells grown in a medium containing d-threonine as the sole nitrogen source produce an enzyme that catalyzes epimerization of threonine. proton nuclear magnetic resonance analysis of the enzyme reaction in deuterium oxide clearly showed epimerization from l- to d-allo-threonine and also from d- to l-allo-threonine. this is the first example of an enzyme that was clearly shown to epimerize threonine. the enzyme has been purified to homogeneity, whic ... | 1993 | 8320235 |
| degradative capability of pseudomonas putida on acetonitrile. | pseudomonas putida, capable of utilizing acetonitrile as a sole source of carbon and nitrogen, was isolated from contaminated soil and water samples collected from industrial sites. the p. putida cells were immobilized in calcium alginate beads. the degradation of acetonitrile by the immobilized cells of p. putida was investigated. the immobilized cells degraded different concentrations of acetonitrile into ammonia and carbon dioxide. the effect of aeration on the degradation rate was also studi ... | 1993 | 8323268 |
| expression of the agga locus of pseudomonas putida in vitro and in planta as detected by the reporter gene, xyle. | in vitro agglutinability by pseudomonas putida, isolate corvallis, with a plant root surface agglutinin is correlated with rapid adhesion of cells of the fluorescent pseudomonad to bean (phaseolus vulgaris) root surfaces. agglutinability in p. putida cells is regulated by nutrient status as well as growth phase. cells grown in three different nutrient complex media are agglutinable at early and mid-late logarithmic phase but become nonagglutinable at stationary phase. cells grown in a minimal me ... | 1993 | 8324250 |
| plasmid-mediated mineralization of naphthalene, phenanthrene, and anthracene. | the well-characterized plasmid-encoded naphthalene degradation pathway in pseudomonas putida ppg7(nah7) was used to investigate the role of the nah plasmid-encoded pathway in mineralizing phenanthrene and anthracene. three pseudomonas strains, designated 5r, dfc49, and dfc50, were recovered from a polynuclear aromatic hydrocarbon-degrading inoculum developed from a manufactured gas plant soil slurry reactor. plasmids pka1, pka2, and pka3, approximately 100 kb in size, were isolated from these st ... | 1993 | 8328809 |
| complete nucleotide sequence of the 5-exo-hydroxycamphor dehydrogenase gene on the cam plasmid of pseudomonas putida (atcc 17453). | we determined the complete nucleotide sequence of the first gene of pseudomonas putida cytochrome p-450cam hydroxylase operon, camd, which encodes 5-exo-hydroxycamphor dehydrogenase. this dehydrogenase apparently consists of 361 amino acids and has a molecular mass of 38.4 kda. structural relationships to other zinc-containing alcohol dehydrogenases also became evident. | 1993 | 8334169 |
| cloning and nucleotide sequence analysis of the ferripyoverdine receptor gene fpva of pseudomonas aeruginosa. | pseudomonas aeruginosa k437 lacks the ferripyoverdine receptor and, as a result, grows poorly on an iron-deficient minimal medium supplemented with ethylenediamine-di(o-hydroxyphenylacetic acid) (eddha) and pyroverdine. by using a phagemid-based in vivo cloning system, attempts were made to clone the receptor gene by complementing this growth defect. several recombinant phagemids carrying p. aeruginosa chromosomal dna which provided for good growth on eddha-pyoverdine-containing medium and which ... | 1993 | 8335619 |
| in vivo reactivation of catechol 2,3-dioxygenase mediated by a chloroplast-type ferredoxin: a bacterial strategy to expand the substrate specificity of aromatic degradative pathways. | the meta-cleavage operon of the tol plasmid pww0 of pseudomonas putida contains 13 genes responsible for the oxidation of benzoate and toluates to krebs cycle intermediates via estradiol (meta) cleavage of (methyl)catechol. the functions of all the genes are known with the exception of xylt. we constructed pww0 mutants defective in the xylt gene, and found that these mutants were not able to grow on p-toluate while they were still capable of growing on benzoate and m-toluate. in the xylt mutants ... | 1993 | 8344270 |
| the pseudomonas putida ml2 plasmid-encoded genes for benzene dioxygenase are unusual in codon usage and low in g+c content. | benzene dioxygenase, catalyzing the oxidation of benzene to cis-1,2-dihydroxy-cyclohexa-3,5-diene, comprises four polypeptides that are encoded by plasmid phmt112 of pseudomonas putida ml2. in this study, the nucleotide (nt) sequences of four genes encoding this enzyme (bedc1c2ba) were determined, and the amino acid (aa) sequences were deduced. the sequence showed significant homology with the chromosomally encoded benzene dioxygenase and toluene dioxygenase genes (73-77% for nt and 83-99% for a ... | 1993 | 8344526 |
| gene components responsible for discrete substrate specificity in the metabolism of biphenyl (bph operon) and toluene (tod operon). | bph operons coding for biphenyl-polychlorinated biphenyl degradation in pseudomonas pseudoalcaligenes kf707 and pseudomonas putida kf715 and tod operons coding for toluene-benzene metabolism in p. putida f1 are very similar in gene organization as well as size and homology of the corresponding enzymes (g. j. zylstra and d. t. gibson, j. biol. chem. 264:14940-14946, 1989; k. taira, j. hirose, s. hayashida, and k. furukawa, j. biol. chem. 267:4844-4853, 1992), despite their discrete substrate rang ... | 1993 | 8349562 |
| hydroxylation and biodegradation of 6-methylquinoline by pseudomonads in aqueous and nonaqueous immobilized-cell bioreactors. | selective culturing of pseudomonads that could degrade quinoline led to enrichment cultures and pure cultures with expanded substrate utilization and transformation capabilities for substituted quinolines in immobilized and batch cultures. immobilized cells of the pseudomonad cultures rapidly transformed quinolines to hydroxyquinolines in bioreactors and were able to tolerate higher substrate concentrations compared with batch cultures. after prolonged incubation on a mixture of quinoline and 6- ... | 1993 | 8357249 |
| indirect utilization of the phytosiderophore mugineic acid as an iron source to rhizosphere fluorescent pseudomonas. | the phytosiderophore mugineic acid (ma) was studied as a source of iron for rhizosphere fluorescent pseudomonads. 55fe supplied as fe-ma was taken up by pseudomonas putida wcs358, b10 and st3 grown under iron deficient conditions. the uptake decreased when the bacteria were grown in the presence of iron. however, no differences in uptake were observed when a siderophore deficient mutant was tested. since ligand exchange between pseudobactin and ma was shown to occur rapidly with a half-life of 2 ... | 1993 | 8358206 |
| reinvestigation of the role of thiol groups of glyoxalase i purified from yeast (saccharomyces cerevisiae). | glyoxalase i has been purified to homogeneity from saccharomyces cerevisiae and tested with two different thiol reagents, i.e., 5,5'-dithiobis-(2-nitrobenzoic acid) (dtnb) and 1-chloro-2,4-dinitrobenzene (cdnb). dtnb reacts with four thiol groups per molecule of enzyme and leads to a complete inhibition which is not reversed by addition of the disulfide-reducing agent dithiothreitol. on the other hand, cdnb slightly affects the glyoxalase-i activity and alkylates only one thiol residue/enzyme. i ... | 1993 | 8373819 |
| regulation of the pcaij genes for aromatic acid degradation in pseudomonas putida. | six of the genes encoding enzymes of the beta-ketoadipate pathway for benzoate and 4-hydroxybenzoate degradation in pseudomonas putida are organized into at least three separate transcriptional units. as an initial step to defining this pca regulon at the molecular level, lacz fusions were made with the pcai and pcaj genes, which encode the two subunits of beta-ketoadipate:succinyl-coenzyme a transferase, the enzyme catalyzing the next-to-last step in the beta-ketoadipate pathway. fusion analyse ... | 1993 | 8376330 |
| isolation and preliminary characterization of the subunits of the terminal component of naphthalene dioxygenase from pseudomonas putida ncib 9816-4. | the terminal oxygenase component (ispnap) of naphthalene dioxygenase from pseudomonas putida ncib 9816-4 was purified to homogeneity. the protein contained approximately 4 g-atoms each of iron and acid-labile sulfide per mol of ispnap, and enzyme activity was stimulated significantly by addition of exogenous iron. the large (alpha) and small (beta) subunits of ispnap were isolated by two different procedures. the nh2-terminal amino acid sequences of the alpha and beta subunits were identical to ... | 1993 | 8376335 |
| cloning and expression of a member of a new cytochrome p-450 family: cytochrome p-450lin (cyp111) from pseudomonas incognita. | cytochrome p-450lin catalyzes the 8-methyl hydroxylation of linalool as the first committed step of its utilization by pseudomonas incognita as the sole carbon source. by using a polymerase chain reaction-based cloning strategy, a 2.1-kb dna fragment containing the cytochrome p-450lin gene (linc) was isolated. an open reading frame of 406 amino acids has been identified as that of p-450lin on the basis of amino acid sequence data from peptides of the native protein. heterologous expression of fu ... | 1993 | 8376348 |
| nicotinoprotein [nad(p)-containing] alcohol/aldehyde oxidoreductases. purification and characterization of a novel type from amycolatopsis methanolica. | extracts of gram-positive bacteria like rhodococcus rhodochrous, rhodococcus erythropolis and amycolatopsis methanolica, but not those of several gram-negative ones, showed dehydrogenase activity for ethanol as well as for methanol when 4-nitroso-n,n-dimethylaniline (ndma) was used as electron acceptor. chromatography of extracts of the first two organisms revealed one activity for both substrates, that of a. methanolica two activities, one of which is able to oxidize methanol and has been purif ... | 1993 | 8385013 |
| energy conservation by pyrroloquinoline quinol-linked xylose oxidation in pseudomonas putida nctc 10936 during carbon-limited growth in chemostat culture. | when grown in carbon source-limited chemostat cultures with lactate or glucose as the carbon and energy source and xylose as an additional source of reducing equivalents. pseudomonas putida nctc 10936 oxidized xylose to xylonolactone and xylonate. no other products were formed from this pentose, nor was it incorporated into biomass. the presence of xylose in these cultures resulted in higher yglucose and ylactate values as compared to cultures without xylose indicating that biologically useful e ... | 1993 | 8385642 |
| in-vivo-generated fusion promoters in pseudomonas putida. | plasmid pest1463 carrying the promoterless pheba operon was cloned into pseudomonas putida paw85, and phenol-utilizing colonies were isolated on minimal plates containing phenol as the only carbon and energy source. in these clones, chromosomally located tn4652 was transposed upstream from the coding sequencing of phea (encoding phenol monooxygenase). sequence analysis together with mapping of the transcription start point of the pheba operon in the recombinant plasmids revealed that fusions of ... | 1993 | 8387446 |
| preelectrophoresis of agarose plugs containing bacterial chromosomal dna prepared for analysis by pulsed field gel electrophoresis can improve the clarity of restriction patterns. | pulsed field gel electrophoresis has indicated that chromosomal dna isolated from stationary phase pseudomonas fluorescens, pseudomonas putida, and escherichia coli cells immobilized in agarose can be fragmented during its release. for p. fluorescens it was demonstrated that the entire chromosome is affected and that there are no specifically fragile sites. the extent of the damage increased both during storage of the dna and also when magnesium ions were provided, suggesting that nucleases may ... | 1993 | 8387734 |
| identification and characterization of the pupb gene encoding an inducible ferric-pseudobactin receptor of pseudomonas putida wcs358. | pseudomonas putida wcs358 can transport iron complexed to a wide variety of pseudobactins produced by other pseudomonas strains. the pupb gene encoding an outer membrane ferric-pseudobactin receptor was isolated from a genomic library of p. putida wcs358. the pupb receptor facilitated iron transport via two distinct heterologous siderophores, i.e. pseudobactin bn8 and pseudobactin bn7. the amino acid sequence deduced from the nucleotide sequence consisted of 804 amino acids (molecular weight 88, ... | 1993 | 8392140 |
| engineering of alkyl- and haloaromatic-responsive gene expression with mini-transposons containing regulated promoters of biodegradative pathways of pseudomonas. | four recombinant mini-tn5 transposons are described which contain outward-facing pm, pu or psal promoters from the catabolic plasmids tol and nah of pseudomonas putida, along with their cognate wild-type regulatory genes (xyls, xylr, nahr) or mutant varieties (xyls2). transcription from such promoters is activated when the host bacteria encounters certain aromatic compounds, such as alkyl- and halobenzoates (xyls, xyls2), alkyl- and halotoluenes (xylr) or salicylates (nahr). these transposons en ... | 1993 | 8393826 |
| essential role of the arg112 residue of cytochrome p450cam for electron transfer from reduced putidaredoxin. | cytochrome p450cam (cyp101) of pseudomonas putida ppg1 in which arg112 is substituted by cys was isolated by in vitro random mutagenesis of the camc gene dna coding for p450cam. the absorption spectra of the purified mutant enzyme were similar to those of the wild type enzyme, but its substrate-dependent nadh oxidation activity in the presence of putidaredoxin (pd) and putidaredoxin reductase (pdr) was extremely low. the rate constant of electron transfer from reduced pd to the heme of the mutan ... | 1993 | 8405387 |
| transformation of 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8- dihydroxyquinoline by quinoline-grown resting cells of pseudomonas putida 86. | resting cells of pseudomonas putida strain 86 were grown on quinoline transformed 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8-dihydroxyquinoline which was not converted further. 7,8-dioxygenating activity was present when the enzymes of quinoline catabolism were induced. quinoline-grown cells of strain 86 treated simultaneously with 2-chloroquinoline and d-(-)-threo-chloramphenicol to prevent protein biosynthesis also formed the cis-7,8-dihydrodiol of 2-chloroquinoline. succinate-grown res ... | 1993 | 8405957 |
| cbbr, a lysr-type transcriptional activator, is required for expression of the autotrophic co2 fixation enzymes of xanthobacter flavus. | xanthobacter flavus is able to grow autotrophically with the enzymes of the calvin cycle for the fixation of co2, which are specified by the cbblsxfp gene cluster. previously, the 5' end of an open reading frame (cbbr), displaying a high sequence similarity to the lysr family of regulatory proteins and transcribed divergently from cbblsxfp, was identified (w. g. meijer, a. c. arnberg, h. g. enequist, p. terpstra, m. e. lidstrom, and l. dijkhuizen, mol. gen. genet. 225:320-330, 1991). this paper ... | 1993 | 8407781 |
| cloning, sequencing, and expression of the pseudomonas putida protocatechuate 3,4-dioxygenase genes. | the genes that encode the alpha and beta subunits of protocatechuate 3,4-dioxygenase (3,4-pcd [ec 1.13.11.3]) were cloned from a pseudomonas putida (formerly p. aeruginosa) (atcc 23975) genomic library prepared in lambda phage. plaques were screened by hybridization with degenerate oligonucleotides designed using known amino acid sequences. a 1.5-kb smai fragment from a 15-kb primary clone was subcloned, sequenced, and shown to contain two successive open reading frames, designated pcah and pcag ... | 1993 | 8407791 |
| heteronuclear nmr analysis of unsaturated fatty acids in poly(3-hydroxyalkanoates). study of beta-oxidation in pseudomonas putida. | poly(3-hydroxyalkanoates) (phas) were isolated from pseudomonas putida kt2442 cultivated on petroselenic acid, oleic acid, and linoleic acid to study beta-oxidation of unsaturated fatty acids. both saturated and unsaturated medium chain length 3-hydroxy fatty acids were found to be constituents of these polymers. with the aid of proton-detected multiple quantum coherence and proton-detected multiple bond coherence nmr spectra the structures of the unsaturated monomers were identified as 3-hydrox ... | 1993 | 8416939 |
| nucleotide sequence and initial functional characterization of the clcr gene encoding a lysr family activator of the clcabd chlorocatechol operon in pseudomonas putida. | the 3-chlorocatechol operon clcabd is central to the biodegradative pathway of 3-chlorobenzoate. the clcr regulatory gene, which activates the clcabd operon, was cloned from the region immediately upstream of the operon and was shown to complement an insertion mutation for growth on 3-chlorobenzoate. clcr activated the clca promoter, which controls expression of the clcabd operon, in trans by 14-fold in an in vivo promoter probe assay in pseudomonas putida when cells were incubated with 15 mm 3- ... | 1993 | 8419291 |
| cloning, sequencing, and genetic characterization of regulatory genes, rina and rinb, required for the activation of staphylococcal phage phi 11 int expression. | the int gene of staphylococcal bacteriophage phi 11 is the only viral gene responsible for the integrative recombination of phi 11. to study the regulation of int gene expression, we determined the 5' end of the transcript by s1 mapping. the presumed promoter is located just 22 nucleotides upstream of the int open reading frame in a region which is conserved between phi 11 and a closely related staphylococcal phage, l54a. to clone the possible regulatory gene, a vector which contained the report ... | 1993 | 8432703 |
| proteins induced by sulfate limitation in escherichia coli, pseudomonas putida, or staphylococcus aureus. | two-dimensional gel electrophoresis of proteins from escherichia coli, pseudomonas putida, and staphylococcus aureus, grown with methionine or one of a variety of organosulfates and organosulfonates as the sole source of sulfur, showed expression of specific sets of 7 to 14 proteins which were not observed during growth with sulfate or cysteine for all three species or with thiocyanate for p. putida and s. aureus. under the same conditions, arylsulfatase activity in p. putida and s. aureus was s ... | 1993 | 8432711 |
| identification and characterization of the exbb, exbd and tonb genes of pseudomonas putida wcs358: their involvement in ferric-pseudobactin transport. | catechol-cephalosporins are siderophore-like antibiotics which are taken up by cells of pseudomonas putida wcs358 via the ferric-siderophore transport pathway. mutants of strain wcs358 were isolated that are resistant to high concentrations of these antibiotics. these mutants failed to grow under iron-limiting conditions, and could not utilize different ferric-siderophores. the mutants fall in three complementation groups. the nucleotide sequence determination identified three contiguous open re ... | 1993 | 8437515 |
| precise deletions in large bacterial genomes by vector-mediated excision (vex). the trfa gene of promiscuous plasmid rk2 is essential for replication in several gram-negative hosts. | we have developed a simple and efficient method of vector-mediated excision (vex) for in vivo generation of precisely defined deletions in large bacterial genomes. the system uses homologous recombination with small cloned fragments on specialized pvex plasmids to insert directly repeated bacteriophage p1 loxp sites at positions flanking the region to be deleted. in the presence of cre recombinase, the loxp sites are efficiently recombined to produce the deletion. deletion endpoints can be direc ... | 1993 | 8450534 |
| nucleotide sequence and over-expression of morphine dehydrogenase, a plasmid-encoded gene from pseudomonas putida m10. | pseudomonas putida m10 was originally isolated from factory waste liquors by selection for growth on morphine. the nadp(+)-dependent morphine dehydrogenase that initiates morphine catabolism is encoded by a large plasmid of 165 kb. treatment of p. putida m10 with ethidium bromide led to the isolation of a putative plasmid-free strain that was incapable of growth on morphine. the structural gene for morphine dehydrogenase, mora, has been located on the plasmid by oligonucleotide hybridization, by ... | 1993 | 8452544 |
| increased expression of the plasmid-determined 2,3-dihydroxybiphenyl dioxygenase gene in strains of escherichia coli, pseudomonas putida and pseudomonas aeruginosa. | a 6.5-kb ecori fragment containing the gene encoding 2,3-dihydroxybiphenyl dioxygenase from the plasmid pbs312 was cloned into broad host range plasmid rsf1010 and expressed in escherichia coli, pseudomonas putida and pseudomonas aeruginosa strains. the increased expression of the gene was orientation-dependent and probably due to the transcription read through from the streptomycin promoter of the vector. subcloning experiments of the psti fragments of pbs312 plasmid using vector pbr322 reveale ... | 1993 | 8454186 |
| copper accumulation by a strain of pseudomonas putida. | a study on the copper accumulation by resting cells of copper-resistant bacteria, isolated from activated sludge and electroplating effluent, was conducted. the best selected strain, identified as pseudomonas putida ii-11, retained copper ions, cu(ii), as high as 6.5% of its dry weight. bacterial cells grown in the sulphate-limiting medium had the highest copper removal capacity [rc, mg of cu(ii)/g of dry cells], while the presence of glucose or sodium azide did not affect cu(ii) rc of the bacte ... | 1993 | 8459779 |
| construction of a pseudomonas hybrid strain that mineralizes 2,4,6-trinitrotoluene. | a bacterium, pseudomonas sp. strain c1s1, able to grow on 2,4,6-trinitrotoluene (tnt), 2,4- and 2,6-dinitrotoluene, and 2-nitrotoluene as n sources, was isolated. the bacterium grew at 30 degrees c with fructose as a c source and accumulated nitrite. through batch culture enrichment, we isolated a derivative strain, called pseudomonas sp. clone a, which grew faster on tnt and did not accumulate nitrite in the culture medium. use of tnt by these two strains as an n source involved the successive ... | 1993 | 8468288 |
| purification of pseudomonas putida acyl coenzyme a ligase active with a range of aliphatic and aromatic substrates. | acyl coenzyme a (acyl-coa) ligase (acyl-coa synthetase [acoas]) from pseudomonas putida u was purified to homogeneity (252-fold) after this bacterium was grown in a chemically defined medium containing octanoic acid as the sole carbon source. the enzyme, which has a mass of 67 kda, showed maximal activity at 40 degrees c in 10 mm k2po4h-napo4h2 buffer (ph 7.0) containing 20% (wt/vol) glycerol. under these conditions, acoas showed hyperbolic behavior against acetate, coa, and atp; the kms calcula ... | 1993 | 8476289 |
| enhanced mineralization of polychlorinated biphenyls in soil inoculated with chlorobenzoate-degrading bacteria. | an altamont soil containing no measurable population of chlorobenzoate utilizers was examined for the potential to enhance polychlorinated biphenyl (pcb) mineralization by inoculation with chlorobenzoate utilizers, a biphenyl utilizer, combinations of the two physiological types, and chlorobiphenyl-mineralizing transconjugants. biphenyl was added to all soils, and biodegradation of 14c-aroclor 1242 was assessed by disappearance of that substance and by production of 14co2. mineralization of pcbs ... | 1993 | 8476293 |
| identification of a cis-acting sequence within the pm promoter of the tol plasmid which confers xyls-mediated responsiveness to substituted benzoates. | the dna sequences within the pm promoter/operator region of the meta operon of the tol plasmid of pseudomonas putida, which confer xyls-mediated responsiveness to substituted benzoates, have been identified through deletion analysis and mutagenesis of the region. integrity and proper phasing of two homologous tandem sequences 5'-tgcaapuaapu-pyggnta-3', separated by six base-pairs and overlapping with the -35 region of the pm promoter, was essential for m-toluate activation of a pm-lacz fusion in ... | 1993 | 8478926 |
| sequences of genes encoding naphthalene dioxygenase in pseudomonas putida strains g7 and ncib 9816-4. | the multicomponent enzyme, naphthalene dioxygenase, initiates the metabolism of naphthalene by pseudomonas putida strains g7 (ppg7) and ncib 9816-4 (pp9816-4). the genes involved (nahaaabacad) are encoded by the nah7 and pdtg1 plasmids, respectively, and form part of the nah operon. the locations of the structural genes were determined on previously cloned fragments of dna. the nucleotide (nt) sequences were determined for nahaaab from pp9816-4 and for nahaaabacad from ppg7. the appropriate open ... | 1993 | 8486285 |
| the amino acid sequence of pseudomonas putida azurin. | the low molecular weight "blue" copper protein, azurin, has been purified from pseudomonas putida (ncib 9869) to homogeneity using various chromatographic techniques including reverse-phase hplc. the amino acid sequence of the n-terminus of the reduced and carboxymethylated protein yielded a single sequence corresponding to aeckv. the complete sequence, comprising 128 amino acid residues with a c-terminal sequence corresponding to tvtlk, was determined from the peptides obtained from a staphyloc ... | 1993 | 8489263 |
| oxidation of aniline to nitrobenzene by nonheme bromoperoxidase. | nonheme bromoperoxidase found in pseudomonas putida catalyzed the bromination of aniline with hydrogen peroxide and bromide ions to give o- and p-bromoanilines. however, in the absence of bromide ions, it oxidized aniline via azobenzene and azoxybenzene finally into nitrobenzene. this is the first report of the biological oxidation of an arylamine to the corresponding nitrocompound at the enzyme level. in addition, nitrobenzene was not formed by a nonheme bromoperoxidase from corallina pilulifer ... | 1993 | 8490583 |
| kinetic studies on benzyl alcohol dehydrogenase encoded by tol plasmid pwwo. a member of the zinc-containing long chain alcohol dehydrogenase family. | the nucleotide sequence of the structural gene for benzyl alcohol dehydrogenase encoded by tol plasmid pwwo of pseudomonas putida has been determined. benzyl alcohol dehydrogenase is a member of the long-chain zinc alcohol dehydrogenase family and, like other alcohol dehydrogenases of this family, contains two zinc atoms per subunit. benzyl alcohol dehydrogenase, while sharing 31% identical residues with horse liver alcohol dehydrogenase, contains several amino acid substitutions near the active ... | 1993 | 8496150 |
| molecular analysis of the plasmid-borne bed gene cluster from pseudomonas putida ml2 and cloning of the cis-benzene dihydrodiol dehydrogenase gene. | pseudomonas putida ml2 contains a large catabolic plasmid, phmt112, carrying genes that encode the dioxygenase and dehydrogenase involved in the catabolism of benzene via the ortho or beta-ketoadipate pathway. phmt112 was derived from a larger and less stable plasmid in p. putida ml2 following growth on succinate as carbon and energy source but was, however, stably maintained in p. putida even in the absence of selection for growth on benzene. cleavage sites for the restriction endonucleases dra ... | 1993 | 8500007 |
| superoxide dismutase activity in root-colonizing pseudomonads. | several saprophytic fluorescent pseudomonads that are aggressive root colonizers express similar specific activities of superoxide dismutase during growth in liquid culture. the pseudomonads have the potential to produce hydrogen peroxide sensitive and hydrogen peroxide insensitive isoforms of superoxide dismutase with distinct mobilities in nondenaturing polyacrylamide gel electrophoresis. synthesis of the hydrogen peroxide insensitive form is enhanced by limited iron availability, by exposure ... | 1993 | 8500011 |
| creatinase in its collapsed a state shows properties of a molten globule with dimeric quaternary structure. | in the past, the molten globule state at acidic ph (a state) has mainly been observed for small single-domain proteins. for more complex proteins such as immunoglobulin, alternatively folded states, with certain characteristics of the molten globule but different thermodynamic properties, were observed. in the present work, the acid-induced structural characteristics of a homodimer, creatinase from pseudomonas putida, are described. the 91-kda protein at ph 2 shows molten globule behavior in tha ... | 1993 | 8504814 |
| physical organization of the upper pathway operon promoter of the pseudomonas tol plasmid. sequence and positional requirements for xylr-dependent activation of transcription. | the upper pathway operon of the pseudomonas putida tol plasmid belongs to the -12/-24 class of promoters. these promoters exhibit three regions critical for regulated transcription, namely, the -12/-24 site for rna polymerase/sigma 54 binding, the -55/-67 region for ihf protein binding, and the -130(uas2)/-170(uas1) region, where two sites for xylr binding are located. the xylr-protected g residues located at -131, -139, -160 and -169 were replaced with as, and the activity of the mutant promote ... | 1993 | 8510657 |
| comparison of the nucleotide sequences of the meta-cleavage pathway genes of tol plasmid pww0 from pseudomonas putida with other meta-cleavage genes suggests that both single and multiple nucleotide substitutions contribute to enzyme evolution. | tol plasmid pww0 from pseudomonas putida mt-2 encodes catabolic enzymes required for the oxidation of toluene and xylenes. the structural genes for these catabolic enzymes are clustered into two operons, the xylcmabn operon, which encodes a set of enzymes required for the transformation of toluene/xylenes to benzoate/toluates, and the xylxyzltegfjqkih operon, which encodes a set of enzymes required for the transformation of benzoate/toluates to krebs cycle intermediates. the latter operon can be ... | 1993 | 8510667 |
| diketocamphane enantiomer-specific 'baeyer-villiger' monooxygenases from camphor-grown pseudomonas putida atcc 17453. | pseudomonas putida atcc 17453 grew with either (+)- or (-)-camphor as sole carbon source. enantiomer-specific 'biological baeyer-villiger' monooxygenases were synthesized irrespective of the camphor isomer used for growth. the two enzymes are probably the products of separate genes but showed many similarities. each consisted of two electrophoretically identical subunits, bound flavin mononucleotide (fmn) non-covalently and accepted electrons from an induced nadh dehydrogenase which interacted w ... | 1993 | 8515237 |
| purification of active e1 alpha 2 beta 2 of pseudomonas putida branched-chain-oxoacid dehydrogenase. | active e1 component of pseudomonas putida branched-chain-oxoacid dehydrogenase was purified from p. putida strains carrying pjrs84 which contains bkdr (encoding the transcriptional activator) and bkda1 and bkda2 (encoding the alpha and beta subunits). expression was inducible, however, 45-, 39- and 37-kda proteins were produced instead of the expected 45-kda and 37-kda proteins. the 45-kda protein was identified as e1 alpha and the 37-kda and 39-kda proteins were identified as separate translati ... | 1995 | 8521848 |
| repression of 4-hydroxybenzoate transport and degradation by benzoate: a new layer of regulatory control in the pseudomonas putida beta-ketoadipate pathway. | pseudomonas putida prs2000 degrades the aromatic acids benzoate and 4-hydroxybenzoate via two parallel sequences of reactions that converge at beta-ketoadipate, a derivative of which is cleaved to form tricarboxylic acid cycle intermediates. structural genes (pca genes) required for the complete degradation of 4-hydroxybenzoate via the protocatechuate branch of the beta-ketoadipate pathway have been characterized, and a specific transport system for 4-hydroxybenzoate has recently been described. ... | 1995 | 8522507 |
| recombination of a 3-chlorobenzoate catabolic plasmid from alcaligenes eutrophus nh9 mediated by direct repeat elements. | alcaligenes eutrophus nh9 was isolated from soil. this strain can utilize 3-chlorobenzoate (3-cb) as a sole source of carbon and energy. most of the 3-cb-negative segregants had lost one of the plasmids present in the parent strain. the genes for catabolism of 3-cb were located within a 9.2-kb saci fragment of this plasmid (penh91). the genes were found to hybridize with genes for components of the modified ortho cleavage pathway from pseudomonas putida. in one of the 3-cb-negative segregants, t ... | 1995 | 8526487 |
| application of reverse transcriptase pcr for monitoring expression of the catabolic dmpn gene in a phenol-degrading sequencing batch reactor. | a modified freeze-thaw method in combination with reverse transcriptase pcr was developed for monitoring gene expression in activated sludge. the sensitivity of the methodology was determined by inoculating non-sterile activated sludge samples with 3-chlorobenzoate-degrading pseudomonas putida ppo301(pro103), which contains the catabolic tfdb gene. tfdb mrna was detected in 10 mg of activated sludge inoculated with 10(4) cfu of the target organism. this technique was subsequently utilized to ana ... | 1995 | 8526513 |
| maintenance and induction of naphthalene degradation activity in pseudomonas putida and an alcaligenes sp. under different culture conditions. | the expression of xenobiotic-degradative genes in indigenous bacteria or in bacteria introduced into an ecosystem is essential for the successful bioremediation of contaminated environments. the maintenance of naphthalene utilization activity is studied in pseudomonas putida (atcc 17484) and an alcaligenes sp. (strain np-alk) under different batch culture conditions. levels of activity decreased exponentially in stationary phase with half-lives of 43 and 13 h for strains atcc 17484 and np-alk, r ... | 1995 | 8526520 |
| coordination of the rieske-type [2fe-2s] cluster of the terminal iron-sulfur protein of pseudomonas putida benzene 1,2-dioxygenase, studied by one- and two-dimensional electron spin-echo envelope modulation spectroscopy. | one- and two-dimensional (1d and 2d) electron spin-echo envelope modulation (eseem) spectroscopy has been used to investigate the ligand environment of the [2fe-2s] cluster from the terminal dioxygenase (ispbed) of the pseudomonas putida benzene dioxygenase complex. the modulation frequencies observed in the 0.5-8.5 mhz region of the fourier transforms of 1d and 2d eseem spectra measured across the electron paramagnetic resonance (epr) absorbance envelope (from gz through to gx) are consistent w ... | 1995 | 8527426 |
| the cytochrome subunit is necessary for covalent fad attachment to the flavoprotein subunit of p-cresol methylhydroxylase. | when p-cresol methylhydroxylase (pcmh) is expressed in its natural host pseudomonas putida, or when the genes of the alpha and beta subunits of the enzyme are expressed together in the heterologous host escherichia coli, flavin-adenine dinucleotide (fad) is covalently attached to tyr384 of the alpha subunit and the correct alpha 2 beta 2 form of the enzyme is assembled. the apoflavoprotein has been expressed in e. coli in the absence of the beta cytochrome c subunit and purified. while noncovale ... | 1995 | 8537385 |
| molecular characterization of the 4-hydroxyphenylacetate catabolic pathway of escherichia coli w: engineering a mobile aromatic degradative cluster. | we have determined and analyzed the nucleic acid sequence of a 14,855-bp region that contains the complete gene cluster encoding the 4-hydroxyphenylacetic acid (4-hpa) degradative pathway of escherichia coli w (atcc 11105). this catabolic pathway is composed by 11 genes, i.e., 8 enzyme-encoding genes distributed in two putative operons, hpabc (4-hpa hydroxylase operon) and hpagedfhi (meta-cleavage operon); 2 regulatory genes, hpar and hpaa; and the gene, hpax, that encodes a protein related to t ... | 1996 | 8550403 |
| bacterial morphinone reductase is related to old yellow enzyme. | morphinone reductase, produced by pseudomonas putida m10, catalyses the nadh-dependent saturation of the carbon-carbon double bond of morphinone and codeinone, and is believed to be involved in the metabolism of morphine and codeine. the structural gene encoding morphinone reductase, designated morb, was cloned from ps. putida m10 genomic dna by the use of degenerate oligonucleotide probes based on elements of the amino acid sequence of the purified enzyme. sequence analysis and structural chara ... | 1995 | 8554504 |
| manganese(ii)-dependent extradiol-cleaving catechol dioxygenase from arthrobacter globiformis cm-2. | a manganese-dependent 3,4-dihydroxyphenylactate 2,3-dioxygenase from arthrobacter globiformis strain cm-2 (mndd) cloned in escherichia coli has been purified to homogeneity. sedimentation equilibrium analysis indicates an alpha 4 homotetrameric holoenzyme structure (4 x 38,861 da). steady-state kinetic analysis of mndd with a variety of substrates and inhibitors yields very similar relative rates to the known fe(ii)- and mn(ii)-dependent 3,4-dihydroxyphenylacetate 2,3-dioxygenases from pseudomon ... | 1996 | 8555170 |
| nucleotide sequence of salicylate hydroxylase gene and its 5'-flanking region of pseudomonas putida kf715. | the salicylate hydroxylase, a flavoprotein monooxygenase, catalyzes the decarboxylative hydroxylation of salicylate to form catechol. nucleotide sequence of a salicylate hydroxylase gene and its 5'-flanking region in chromosomal dna of pseudomonas putida kf715 was analyzed. the salicylate hydroxylase was encoded in an open reading frame with 1308 base pairs which can encode a polypeptide of molecular weight 48 kda with 435 amino acids. the open reading frame was preceded by a putative ribosome-b ... | 1996 | 8561793 |
| natural selection of pah-degrading bacterial guilds at coal-tar disposal sites. | microbial activity patterns at buried coal-tar disposal sites have been under investigation for several years to determine the response of naturally occurring microflora to polycyclic aromatic hydrocarbons (pahs) at the sites. at one site in upstate new york, data have shown enrichment of pah-degrading bacteria in subsurface contaminated zones but not in uncontaminated zones. similar work at a midwestern site showed that the same trends existed in a heterogeneous disposal site except that a bore ... | 1995 | 8565896 |
| recruitment of co-metabolic enzymes for environmental detoxification of organohalides. | polyhalogenated compounds are often environmentally persistent and toxic to mammals. microorganisms that metabolize these compounds can detoxify contaminated environments. different biochemical mechanisms are used to metabolize polyhalogenated compounds, but few naturally occurring bacteria have this capability. a recombinant bacterium was constructed to metabolize polyhalogenated compounds to nonhalogenated end products. seven genes were expressed in pseudomonas putida g786 to biosynthesize cyt ... | 1995 | 8565909 |
| analysis of the rpod gene encoding the principal sigma factor of pseudomonas putida. | the gene (rpod) encoding the principal sigma factor of pseudomonas putida (pp) was cloned and sequenced. the amino-acid sequence deduced from the nucleotide sequence of rpod contained sequences with significant similarity to the conserved region of the principal sigma factors. in vivo transcriptional analyses revealed that the pp rpod is transcribed as a monocistronic mrna of 2.1 kb and that the transcription of rpod is under control of the heat-shock (hs) response. the transcription start point ... | 1995 | 8566819 |
| polymerase chain reaction for verification of fluorescent colonies of erwinia chrysanthemi and pseudomonas putida wcs358 in immunofluorescence colony staining. | the potential of polymerase chain reaction (pcr) for verifying the identity of colonies stained by the immunofluorescence colony-staining (ifc) procedure was investigated. using primers directed against conserved sequences of the pectate lyase-genes coding for isozymes pla, pld and ple of erwinia chrysanthemi, the authors confirmed the identity of 96% of 20 fluorescent target colonies, punched from ifc-stained samples with pure cultures. in pour plates with mixtures of erw. chrysanthemi and non- ... | 1995 | 8567494 |
| cloning and characterization of styrene catabolism genes from pseudomonas fluorescens st. | a gene bank from pseudomonas fluorescens st was constructed in the broad-host-range cosmid plafr3 and mobilized into pseudomonas putida paw340. identification of recombinant cosmids containing the styrene catabolism genes was performed by screening transconjugants for growth on styrene and epoxystyrene. transposon mutagenesis and subcloning of one of the selected genome fragments have led to the identification of three enzymatic activities: a monooxygenase activity encoded by a 3-kb psti-ecori f ... | 1996 | 8572689 |
| nondisruptive detection of activity of catabolic promoters of pseudomonas putida with an antigenic surface reporter system. | a simple procedure to detect the switching on and off of catabolic promoters of pseudomonas putida, at the level of single cells based on the immunodetection of a reporter epitope expressed on the surface of bacterial cells, has been developed. to do this, the antigenic sequence asp-leu-pro-pro-asn-ser-asp-val-val-asp, from a coronavirus, was inserted genetically in the permissive site around amino acid position 153 of the lamb protein (maltose and lambda phage receptor) of escherichia coli. whe ... | 1996 | 8572699 |
| molecular cloning of novel genes for polycyclic aromatic hydrocarbon degradation from comamonas testosteroni gz39. | three strains of comamonas testosteroni were isolated from river sediment for the ability to degrade phenanthrene; two of the strains also grew on naphthalene, and one strain also grew on anthracene. the homology of the genes for polycyclic aromatic hydrocarbon degradation in these strains to the classical genes (nah) for naphthalene degradation from pseudomonas putida ncib 9816-4 was determined. the three c. testosteroni strains showed no homology to the nah gene probe even under low-stringency ... | 1996 | 8572701 |
| purification and characterization of dihydroorotase from pseudomonas putida. | dihydroorotase was purified to homogeneity from pseudomonas putida. the relative molecular mass of the native enzyme was 82 kda and the enzyme consisted of two identical subunits with a relative molecular mass of 41 kda. the enzyme only hydrolyzed dihydro-l-orotate and its methyl ester, and the reactions were reversible. the apparent km and vmax values for dihydro-l-orotate hydrolysis (at ph 7.4) were 0.081 mm and 18 mumol min-1 mg-1, respectively; and those for n-carbamoyl-dl-aspartate (at ph 6 ... | 1995 | 8572888 |
| structure and characterization of isopyoverdin from pseudomonas putida btp1 and its relation to the biogenetic pathway leading to pyoverdins. | pyoverdin type siderophores produced by six fluorescent pseudomonas strains isolated from different rhizospheres were purified and characterized. the purified ferri-pyoverdins were tested for their ability to promote the growth of other strains grown under iron deficiency conditions. only the one obtained from pseudomonas putida btp1 did not act as a growth promoter. the structure of the btp1 siderophore was elucidated by spectroscopic methods and degradation studies. it turned out that it conta ... | 1995 | 8579680 |
| [epidemiology of microbial resistance to biocides]. | in order to estimate the distribution of bacteria and fungi with an elevated level of resistance to antimicrobial substances, we have analyzed water samples and surveyed institutions presumably concerned with analyses of microbial resistance (university institutes for hygiene, health authorities) by means of questionnaire. a total of 41 water samples was drawn from various aquatic biotopes in the region of heidelberg. the samples originated from the effluents of a community sewage treatment plan ... | 1995 | 8579712 |
| structural analysis of the l-methionine gamma-lyase gene from pseudomonas putida. | the gene encoding l-methionine gamma-lyase from pseudomonas putida was cloned and the primary structure of the enzyme was deduced from its nucleotide sequence. the l-methionine gamma-lyase gene was expressed in escherichia coli. the amino acid sequences of brcn-digested peptides agreed with the corresponding parts of the l-methionine gamma-lyase sequence determined from the gene structure. the polypeptide is composed of 398 amino acid residues with a calculated molecular weight of 42,626, corres ... | 1995 | 8586629 |
| the reca protein as a model molecule for molecular systematic studies of bacteria: comparison of trees of recas and 16s rrnas from the same species. | the evolution of the reca protein was analyzed using molecular phylogenetic techniques. phylogenetic trees of all currently available complete reca proteins were inferred using multiple maximum parsimony and distance matrix methods. comparison and analysis of the trees reveal that the inferred relationships among these proteins are highly robust. the reca trees show consistent subdivisions corresponding to many of the major bacterial groups found in trees of other molecules including the alpha, ... | 1995 | 8587109 |