Publications

characterization of the ptfi91-family replicon of thiobacillus ferrooxidans plasmids.	plasmids found in six strains of thiobacillus ferrooxidans were mapped and compared in an effort to detect the origin of replication. four strains yielded an identical 9.8-kb plasmid, ptfi91. restriction mapping and southern blot hybridization analysis were used to confirm this finding. dissimilar plasmids found in two other strains contained a conserved 2.2-kb saci region common to ptfi91. dna sequence analysis of this region showed structural features common to bacterial plasmid replicons. a c ...	1995	8590413
catabolite repression of the toluene degradation pathway in pseudomonas putida harboring pww0 under various conditions of nutrient limitation in chemostat culture.	in earlier studies, the pathway of toluene and m- and p-xylene degradation (tol pathway) in pseudomonas putida (pww0) was found to be subject to catabolite repression when the strain was grown at the maximal rate on glucose or succinate in the presence of an inducer. this report describes catabolite repression of the tol pathway by succinate in chemostat cultures run at a low dilution rate (d = 0.05 h-1) under different conditions of inorganic-nutrient limitation. the activity of benzylalcohol d ...	1996	8593060
flow cytometric detection of specific genes in genetically modified bacteria using in situ polymerase chain reaction.	use of the polymerase chain reaction, coupled with flow cytometry, to detect a plasmid encoded xyle gene sequence in intact cells of escherichia coli and pseudomonas putida was investigated. optimal incorporation of fluorescently labelled dutp into a full length pcr product required substitution at a level of 2:3 dutp:dttp. formaldehyde fixed cells of both species were counted before and after thermal cycling. sufficient numbers of cells remained intact for subsequent detection using microscopy ...	1995	8593955
characterization of type ii protein secretion (xcp) genes in the plant growth-stimulating pseudomonas putida, strain wcs358.	in pseudomonas aeruginosa, the products of the xcp genes are required for the secretion of exoproteins across the outer membrane. despite structural conservation of the xcp components, secretion of exoproteins via the xcp pathway is generally not found in heterologous organisms. to study the specificity of this protein secretion pathway, the xcp genes of another fluorescent pseudomonad, the plant growth-promoting pseudomonas putida strain wcs358, were cloned and characterized. nucleotide sequenc ...	1996	8602167
detection of pseudomonas aeruginosa from ovine fleece washings by pcr amplification of 16s ribosomal rna.	two oligonucleotides were selected from the variable regions of the 16s rrna gene of p. aeruginosa and used as pcr primers for the detection of p. aeruginosa. the specificity of the primers was tested against the following bacterial species; pseudomonas putida, pseudomonas cepacia, xanthamonas maltophilia, pseudomonas mendocina, pseudomonas stutzeri, pseudomonas fluorescens, pseudomonas alcaligenes and pseudomonas diminuta. these primers had a sensitivity of detection of 1 pg of chromosomal dna ...	1995	8604555
a novel cell surface polysaccharide in pseudomonas putida wcs358, which shares characteristics with escherichia coli k antigens, is not involved in root colonization.	previously we have shown that flagella and the o-specific polysaccharide of lipopolysaccharide play a role in colonization of the potato root by plant growth-promoting pseudomonas strains wcs374 and wcs358. in this paper, we describe a novel cell surface-exposed structure in pseudomonas putida wcs358 examined with a specific monoclonal antibody. this cell surface structure appeared to be a polysaccharide, which was accessible to the monoclonal antibody at the outer cell surface. further study re ...	1996	8606170
carbon catabolite repression of phenol degradation in pseudomonas putida is mediated by the inhibition of the activator protein phlr.	enzymes involved in (methyl)phenol degradation of pseudomonas putida h are encoded by the catabolic operon (phla-l) on plasmid ppgh1. transcription of this operon by the sigma54 (rpon)-containing rna polymerase is positively controlled by the gene product of the divergently transcribed phlr in response to the availability of the respective substrate. additionally, phenol degradation is subject to carbon catabolite repression induced by organic acids (e.g., succinate, lactate, and acetate) or car ...	1996	8606180
gene cloning and characterization of pseudomonas putida l-methionine-alpha-deamino-gamma-mercaptomethane-lyase.	methionine dependency has been reported in cancer cell lines and primary tumors. thus, l-methionine deprivation might have potential value for the treatment of human cancers with a methionine requirement. l-methionine-alpha-deamino-gamma-mercaptomethane-lyase has been reported to decrease plasma methionine levels and to inhibit tumor growth in experimental animals but has not been studied extensively because sufficient homogeneous enzyme was not available. in this study, we cloned the l-methioni ...	1996	8616859
quinaldine 4-oxidase from arthrobacter sp. rü61a, a versatile procaryotic molybdenum-containing hydroxylase active towards n-containing heterocyclic compounds and aromatic aldehydes.	quinaldine 4-oxidase from arthrobacter sp. rü61a, an inducible molybdenum-containing hydroxylase, was purified to homogeneity by an optimized five-step procedure. molecular oxygen is proposed as physiological electron acceptor. electrons are also transferred to artificial electron acceptors with e'o > -8 mv. the molybdo-iron/sulfur flavoprotein regiospecifically attacks its n-heterocyclic substrates: isoquinoline and phthalazine are hydroxylated adjacent to the n-heteroatom at cl, whereas quinal ...	1996	8617260
the pseudomonas putida peptidoglycan-associated outer membrane lipoprotein is involved in maintenance of the integrity of the cell cell envelope.	pseudomonas putida 14g-3, a derivative of the natural soil inhabitant p. putida kt2440, exhibited a chromosomal insertion of a mini-tn5/'phoa transposon that resulted in reduced ability to colonize soil. in vitro characterization of p. putida 14g-3 revealed that it exhibited an altered cell morphology and envelope, as revealed by electron microscopy. the derived strain was sensitive to sodium dodecyl sulfate, deoxycholate, and edta, produced clumps when it reached high cell densities in the late ...	1996	8626299
identification of the repressor subdomain within the signal reception module of the prokaryotic enhancer-binding protein xylr of pseudomonas putida.	in the presence of m-xylene, the protein xylr encoded by the tol plasmid of pseudomonas putida, activates the final sigma54-dependent promoter pu. early activation stages involve the release of the intramolecular repression caused by the signal reception n-terminal (a domain) of xylr on the central module of the protein. a genetic approach has been followed to locate the specific segment within a domain of xylr that is directly responsible for its down-regulation in the absence of inducer, as co ...	1996	8626467
identification of a partition and replication region in the alcaligenes eutrophus megaplasmid pmol28.	a 4.64 kb region of the 180 kb heavy metal resistance plasmid pmol28 of alcaligenes eutrophus ch34, previously shown to be able to replicate autonomously, was sequenced and analyzed. three genes involved in plasmid maintenance were identified: para28 and parb28 are involved in plasmid partitioning and stability, while repa28 encodes a protein required for replication. in addition to the par ab28 genes, a third locus, pars28, required in cis active partitioning was identified. the parabs28 locus ...	1996	8628216
p-cumate catabolic pathway in pseudomonas putida fl: cloning and characterization of dna carrying the cmt operon.	pseudomonas putida f1 utilizes p-cumate (p-isopropylbenzoate) as a growth substrate by means of an eight-step catabolic pathway. a 35.75-kb dna segment, within which the cmt operon encoding the catabolism of p-cumate is located, was cloned as four separate overlapping restriction fragments and mapped with restriction endonucleases. by examining enzyme activities in recombinant bacteria carrying these fragments and sub-cloned fragments, genes encoding most of the enzymes of the p-cumate pathway w ...	1996	8631713
identification of tonb homologs in the family enterobacteriaceae and evidence for conservation of tonb-dependent energy transduction complexes.	the transport of fe(iii)-siderophore complexes and vitamin b12 across the outer membrane of escherichia coli requires the tonb-dependent energy transduction system. a set of murine monoclonal antibodies (mabs) was generated against an e. coli trpc-tonb fusion protein to facilitate structure and function studies. in the present study, the epitopes recognized by these mabs were mapped, and their distribution in gram-negative organisms was examined. cross-species reactivity patterns obtained agains ...	1996	8631714
catechol 2,3-dioxygenases functional in oxygen-limited (hypoxic) environments.	we studied the degradation of toluene for bacteria isolated from hypoxic (i.e., oxygen-limited) petroleum-contaminated aquifers and compared such strains with other toluene degraders. three pseudomonas isolates, p. pickettii pko1, pseudomonas sp. strain w31, and p. fluorescens cfs215, grew on toluene when nitrate was present as an alternate electron acceptor in hypoxic environments. we examined kinetic parameters (k(m) and vmax) for catechol 2,3-dioxygenase (c230), a key shared enzyme of the tol ...	1996	8633871
purification of 2,3-dihydroxybiphenyl 1,2-dioxygenase from pseudomonas putida ou83 and characterization of the gene (bphc).	the 2,3-dihydroxybiphenyl 1,2-dioxygenase (2,3-dbpd) of pseudomonas putida ou83 was constitutively expressed and purified to apparent homogeneity. the apparent molecular mass of the native enzyme was 256 kda, and the subunit molecular mass was 32 kda. the data suggested that 2,3-dbpd was an octamer of identical subunits. the nucleotide sequence of a dna fragment containing the bphc region was determined. the deduced protein sequence for 2,3-dbpd consisted of 292 amino acid residues, with a calcu ...	1996	8633883
expression of the tol plasmid xyls gene in pseudomonas putida occurs from a alpha 70-dependent promoter or from alpha 70- and alpha 54-dependent tandem promoters according to the compound used for growth.	growth of pseudomonas putida (pwwo) on alkylbenzoates requires the expression of the meta pathway operon, which is mediated by the xyls protein after binding of a benzoate effector. alternatively, in cells growing on toluene or its aromatic alcohols, overexpression of xyls mediated by xylr activated by these compounds leads to overproduction of the xyls regulator, which even in the absence of benzoate effectors stimulates transcription from the meta cleavage pathway operon promoter. we show here ...	1996	8636038
physical and functional analysis of the prokaryotic enhancer of the sigma 54-promoters of the tol plasmid of pseudomonas putida.	the physical and the functional organization of the upstream cis-acting sequence that controls at a distance the transcriptional activity of pu and ps, the two sigma 54-dependent promoters of the tol (toluene/xylene biodegradation) operons of pseudomonas putida, have been determined. dnase i and hydroxyl radical footprinting of the promoters with the purified and pre-activated enhancer-binding protein xylr clearly indicated the presence of two distinct binding sites (proximal and distal) that we ...	1996	8636992
in vitro activities of an n-terminal truncated form of xylr, a sigma 54-dependent transcriptional activator of pseudomonas putida.	a truncated derivative of the xylr protein, which is able to constitutively activate the sigma 54-dependent pu promoter of the tol (toluene biodegradation) plasmid of pseudomonas putida, has been purified to homogeneity and its various activities have been separately examined, in vitro. the truncated regulator xylr delta a was deleted of the signal reception n-terminal module present in wild-type xylr, but retained its central activation domain and the dna binding segment, located at its c termi ...	1996	8636993
mechanism of the reaction catalyzed by mandelate racemase: structure and mechanistic properties of the d270n mutant.	on the basis of the available high-resolution structures of mandelate racemase (mr) from pseudomonas putida [landro, j.a., gerlt, j.a., kozarich, j.w., koo, c.w., shah, v.j., kenyon, g.l., neidhart, d.j., fujita, j., & petsko, g.a. (1994) biochemistry 33, 635-643], lys 166 and his 297 are positioned appropriately to participate in catalysis as acid/base catalysts, with lys 166 participating as the (s)-specific acid/base catalyst and his 297 participating as the (r)-specific acid/base catalyst. t ...	1996	8639525
untranslated sequence upstream of mara in the multiple antibiotic resistance locus of escherichia coli is related to the effector-binding domain of the xyls transcriptional activator.	mara, the 129-amino-acid (aa) protein which plays a crucial role in the multiple antibiotic resistance (mar) phenotype in escherichia coli, shows homology to members of the xyls/arac family of transcriptional regulators. although these proteins vary in size from around 100 to 350 aa they all contain a dna-binding domain with a helix-turn-helix motif. the larger ones, e.g., xyls, arac, and rob, contain an additional domain either at their amino- or at their carboxyterminus. this domain is importa ...	1996	8642609
muconolactone isomerase of the 3-oxoadipate pathway catalyzes dechlorination of 5-chloro-substituted muconolactones.	an enzyme of alcaligenes eutrophus jmp 134 which catalyzes dechlorination of (4r, 5r)- and (4r,5s)-5-chloro-3-methyl- and (4r, 5s)-5-chloromuconolactone of principally 3-methyl-trans-, 3-methyl-cis-dienelactone and cis-dienelactone, respectively, was purified to homogeneity. the enzyme was identified as muconolactone isomerase on the basis of its high activity with muconolactone and on its high degree of sequence similarity with previously described muconolactone isomerases. molecular mass deter ...	1996	8647072
comparison of the "rieske" [2fe-2s] center in the bc1 complex and in bacterial dioxygenases by circular dichroism spectroscopy and cyclic voltammetry.	two different types of "rieske" [2fe-2s] clusters have been observed in proteins, one in the bc complexes of the respiratory chain and the other in bacterial dioxygenases. we have compared the circular dichroic (cd) spectra and redox properties of the water soluble fragment of the rieske center of the bovine heart mitochondrial bc1 complex (isf) and of the ferredoxin from benzene dioxygenase in pseudomonas putida ml2 (fdbed). spinach ferredoxin was also measured for comparison. the redox potenti ...	1996	8652534
vtr expression cassettes for engineering conditional phenotypes in pseudomonas: activity of the pu promoter of the tol plasmid under limiting concentrations of the xylr activator protein.	a simplified procedure to construct recombinant pseudomonas putida (pp) and related bacteria, which transcribe conditionally specific genes inserted into their chromosome in response to lac inducers such as iptg, has been developed. the method is based on the so-called vtr expression cassettes. these are three small (1.98-kb) dna segments engineered as noti restriction fragments that include a laciq gene along with the hybrid trp/lac promoter, ptrc, followed by an optimised translation initiatio ...	1996	8654996
site-directed mutagenesis of conserved amino acids in the alpha subunit of toluene dioxygenase: potential mononuclear non-heme iron coordination sites.	the terminal oxygenase component of toluene dioxygenase from pseudomonas putida f1 is an iron-sulfur protein (isp(tol)) that requires mononuclear iron for enzyme activity. alignment of all available predicted amino acid sequences for the large (alpha) subunits of terminal oxygenases showed a conserved cluster of potential mononuclear iron-binding residues. these were between amino acids 210 and 230 in the alpha subunit (todc1) of isp(tol). the conserved amino acids, glu-214, asp-219, tyr-221, hi ...	1996	8655491
comamonas testosteroni 3-ketosteroid-delta 4(5 alpha)-dehydrogenase: gene and protein characterization.	comamonas testosteroni delta 4(5 alpha)- and delta1-dehydrogenases [delta4(5alpha)- and delta1dh] are key enzymes in the degradation of steroids having an a:b ring fusion in a trans configuration. we previously reported the isolation of the delta1dh gene (p. plesiat, m. grandguillot, s. harayama, s. vragar, and y. michel briand, j. bacteriol. 173:7219-7227, 1991). in this study, the gene encoding delta 4(5 alpha)dh was cloned in escherichia coli on a 16-kbp bamhi fragment by screening a genomic ...	1996	8655514
oxidation of nitric oxide by a new heterotrophic pseudomonas sp.	a new bacterial strain isolated from soil consumed nitric oxide (no) under oxic conditions by oxidation to nitrate. phenotypic and phylogenetic characterization of the new strain ps88 showed that it represents a previously unknown species of the genus pseudomonas, closely related to pseudomonas fluorescens and pseudomonas putida. the heterotrophic, obligately aerobic strain ps88 was not able to denitrify or nitrify; however, strain ps88 oxidized no to nitrate. no was not reduced to nitrous oxide ...	1996	8661941
biodegradation of mixed wastes in continuously operated cyclic reactors.	the problem of simultaneous biodegradation of two dissimilar substrates in a continuously operated cyclic reactor was studied both at the theoretical and experimental levels using a simple model system. the system involved media containing mixtures of glucose and phenol as carbon sources. a pure culture of pseudomonas putida (atcc 17514) was employed. independent kinetic experiments have revealed that glucose and phenol are involved in a crossinhibitory uncompetitive kinetic interaction. the dyn ...	1996	8669919
stoichiometry of bkdr to substrate dna in pseudomonas putida.	bkdr is the transcriptional activator of the bkd operon of pseudomonas putida, which encodes branched chain keto acid dehydrogenase. bkdr binds to a large region of dna between its own structural gene and the first gene of the bkd operon. the object of the present studies was to determine the stoichiometry of binding as part of an effort to understand the action of bkdr in regulation of the bkd operon. [35s]bkdr was prepared and found to be essentially 100% active in the gel shift assay. only on ...	1996	8670279
characterization and distribution of tartrate utilization genes in the grapevine pathogen agrobacterium vitis.	agrobacterium vitis is a common pathogen of grapevine. most strains utilize tartrate, an abundant compound in grapevine. strain ab3 carries two tartrate utilization (or tar) regions: tar-i (on the large ptrab3 plasmid) and tar-ii (on the ab3 ti plasmid). tar-i and tar-ii were structurally and functionally analyzed and are similar to the tar-iii region from the tartrate utilization plasmid ptrab4 of the nopaline-type a. vitis strain ab4 (crouzet and otten, j. bacteriol. 1995, 177:6518-6526). the ...	1996	8672817
the purification and crystallisation of 2,5-diketocamphane 1,2-monooxygenase and 3,6-diketocamphane 1,6-monooxygenase from pseudomonas putida ncimb 10007.		1996	8674690
structure of chromosomal dna coding for pseudomonas putida s-1 salicylate hydroxylase.	a gene coding for the salicylate hydroxylase has been isolated from chromosomal dna of pseudomonas putida s-1 and sequenced. the dna fragment contained an open reading frame of 1266 bp encoding a polypeptide of 421 amino acid residues. the predicted amino acid sequence of the protein gave a good agreement with the sequences determined with the peptides isolated from the enzyme but methionine residue in the amino terminal was deleted in the n-terminal sequence of the enzyme protein. the nucleotid ...	1996	8695632
chloroform mineralization by toluene-oxidizing bacteria.	seven toluene-oxidizing bacterial strains (pseudomonas mendocina kr1, burkholderia cepacia g4, pseudomonas putida f1, pseudomonas pickettii pko1, and pseudomonas sp. strains envpc5, envbf1, and env113) were tested for their ability to degrade chloroform (cf). the greatest rate of cf oxidation was achieved with strain envbf1 (1.9 nmol/min/mg of cell protein). cf also was oxidized by p. mendocina kr1 (0.48 nmol/min/mg of cell protein), strain envpc5 (0.49 nmol/min/mg of cell protein), and escheric ...	1996	8702263
stereospecificity of thermostable ornithine 5-aminotransferase for the hydrogen transfer in the l- and d-ornithine transamination.	the thermostable ornithine 5-aminotransferase of a thermophile, bacillus sp. ym-2, is unique in acting on both enantiomers of ornithine, although less effectively on the d-enantiomer. we studied the stereospecificity of the enzyme for the hydrogen abstraction from c-5 of the substrate moiety and the addition and removal of the hydrogen at c-4' of the cofactor (pyridoxal phosphate and pyridoxamine phosphate) moiety of the external schiff base intermediate in the transamination of l- and d-ornithi ...	1996	8703952
the pseudomonas aeruginosa tonb gene encodes a novel tonb protein.	the pseudomonas aeruginosa tonb gene was cloned by complementation of the tonb mutation of pseudomonas putida strain te516 (w. bitter, j. tommassen & p.j. weisbeek, 1993, mol microbiol 7, 117-130). the gene was 1025 bp in length, capable of encoding a protein of 36860 da. as with previously described tonb proteins, the p. aeruginosa tonb (tonbp.a.) was rich in pro residues (18.1%) and contained glu-pro/lys-pro repeats. unlike previously described tonb proteins, however, tonbp.a. lacked an n-term ...	1996	8704984
atp binding to the sigma 54-dependent activator xylr triggers a protein multimerization cycle catalyzed by uas dna.	the events that take place at the prokaryotic enhancer of the pu promoter of pseudomonas putida prior to the engagement of the sigma 54-rna polymerase (sigma 54-rnap) have been studied in vitro. atp hydrolysis by xylr, the cognate regulator of the system, is preceded by the multimerization of xylr at the enhancer, which is itself triggered by the sole allosteric effect of atp binding to the protein. since adp is unable to support multimerization, atp hydrolysis might be followed by a return to t ...	1996	8706137
a transposon for green fluorescent protein transcriptional fusions: application for bacterial transport experiments.	the movement of bacteria through groundwater is a poorly understood process. factors such as soil porosity and mineralogy, heterogeneity of soil particle size, and response of the bacteria to their environment contribute to the pattern of bacterial flow. the identification of transported bacteria is often a limiting factor in both laboratory and field transport experiments. two bacterial strains were modified for use in bacterial transport experiments: a strain of escherichia coli harboring the ...	1996	8707057
bacterial plasmid conjugation on semi-solid surfaces monitored with the green fluorescent protein (gfp) from aequorea victoria as a marker.	horizontal transfer of the tol plasmid was examined in pseudomonas putida (pp) kt2442 micro-colonies on semi-solid agar surfaces. horizontal gene transfer is usually studied in large populations where all information is based on average estimates of the transfer events in the entire population. we have used the green fluorescent protein (gfp) from the jellyfish aequorea victoria as a plasmid marker, in combination with single-cell observations. this provided hitherto unknown details on the distr ...	1996	8707058
structure of catechol 2,3-dioxygenase gene encoded in chromosomal dna of pseudomonas putida kf715.	a catechol 2,3-dioxygenase gene in chromosomal dna of p. putida kf715 was cloned and its nucleotide sequence analyzed. the enzyme gene was composed of 924 base pairs with atg initiation codon and tga termination codon, which can encode a polypeptide of molecular weight 35 kda containing 307 amino acids. a promoter-like sequence and a ribosome-binding sequence were identified upstream the enzyme gene. a deduced amino acid sequence of the catechol 2,3-dioxygenase exhibited 94% identity with that o ...	1996	8713131
[selection and properties of totally phage-resistant mutant pseudomonas putida ppg1].	the efficiency of using bacteria in open systems to degrade different anthropogenic toxic pollutants can depend strongly on the interaction between these bacteria and natural bacteriophages. the possibility of selecting bacterial pseudomonas putida mutants resistant to all bacteriophages of this species known so far was tested (in our work, these mutants were designated totally phage-resistant mutants). in a model experiment, changes in the composition of a population upon prolonged growth of ba ...	1996	8723627
preadapted inocula for limiting the risk of errors in biodegradability tests.	reducing the time for biodegradability tests to 28 days poses a problem when the inoculum contains few biodegraders, as a biodegradable xenobiotic must give a positive result within this time. the influence of initial concentration (x0, number of cells liter-1) on the lag time (hours) of para-nitrophenol biodegradability tests was examined using different concentrations of adapted pseudomonas putida with para-nitrophenol as the sole carbon and energy source. lag time decreased as bacterial densi ...	1996	8727519
tol plasmid transcription factor xyls binds specifically to the pm operator sequence.	xyls, an arac family transcription factor, positively regulates transcription of pseudomonas putida tol plasmid meta operon from the pm promoter. a tandem of 15 bp homologous direct repeats, separated by 6 bp and overlapping with the -35 hexamer of the promoter, is required for the activation of pm by xyls in vivo. in this study we have characterized specific binding of xyls to the pm operator om. xyls was overexpressed with an epitope tag in its n-terminus. tagged xyls (n-xyls) was immunopurifi ...	1996	8736536
organization and expression in pseudomonas putida of the gene cluster involved in cephalosporin biosynthesis from lysobacter lactamgenus yk90.	the lysobacter lactamgenus yk90 pcbab gene encoding delta-(l-alpha-aminoadipyl)-l-cysteinyl-d-valine (acv) synthetase is located immediately upstream of the pcbc gene in the same orientation in the gene cluster involved in cephalosporin biosynthesis. the pcbab gene encodes a large polypeptide composed of 3722 amino acid residues with a molecular mass of 411593 da. the predicted amino acid sequence has a high degree of similarity with those of known acv synthetases from fungi and actinomycetes. w ...	1996	8737573
choice of microbial host for the naphthalene dioxygenase bioconversion.	the use of whole cell biotransformations for single and multistep enzyme conversions is gaining widespread application. in this study the naphthalene dioxygenase nah a gene was transferred into pseudomonas aeruginosa pac 1r, escherichia coli jm107 and pseudomonas putida ppg 277. the effect of ethanol on these genetically engineered gram-negative bacteria was studied by measurement of enzyme activity, stability and cell integrity. ethanol has been used in biotransformations as a co-substrate carb ...	1996	8757940
sigma54-dependent transcription of the pseudomonas putida xyls operon is influenced by the iiantr protein of the phosphotransferase system in escherichia coli.	iiantr, encoded within the rpon operons of many gram-negative bacteria, is a homologue of a class of phosphoryl transfer proteins of the phosphoenolpyruvate: sugar phosphotransferase system. we have used a xyls operon-lacz fusion from the tol plasmid of pseudomonas putida to show that iiantr influences sigma 54-dependent transcription when the xyls operon is expressed in escherichia coli. loss of iiantr influences, but does not abolish cyclic amp-independent carbon catabolite repression.	1996	8761731
engineering hybrid pseudomonads capable of utilizing a wide range of aromatic hydrocarbons and of efficient degradation of trichloroethylene.	we constructed hybrid pseudomonas strains in which the bpha1 gene (coding for a large subunit of biphenyl dioxygenase) is replaced with the todc1 gene (coding for a large subunit of toluene dioxygenase of pseudomonas putida fl) within chromosomal biphenyl-catabolic bph gene clusters. such hybrid strains gained the novel capability to grow on a wide range of aromatic hydrocarbons, and, more interestingly, they degraded chloroethenes such as trichloroethylene and cis-1,2-dichloroethylene very effi ...	1996	8763929
cold shock proteins and cold acclimation proteins in the psychrotrophic bacterium pseudomonas putida q5 and its transconjugant.	the production of cold shock proteins (csps) and cold acclimation proteins (caps) was characterized in the psychrotrophic bacterium pseudomonas putida q5 and its transconjugant p. putida q5t which contains the toluene-degradative tol (pwwo) plasmid, using two-dimensional gel electrophoresis and computing scanning laser densitometry. similar growth rates for the psychrotrophic bacterium p. putida q5 and the transconjugant were found at temperatures ranging from 30 to 0 degree c. sixteen proteins ...	1996	8776850
recruitment and expression of toluene/trichloroethylene biodegradation genes in bacteria native to deep-subsurface sediments.	four plasmids, each encoding a combination of either an escherichia coli or pseudomonas putida promoter and either toluene dioxygenase or toluene monooxygenase, were electroporated into five bacterial strains isolated from sediments found at depths of 91 to 295 m. four of these engineered bacterial strains demonstrated both toluene and trichloroethylene degradation activities.	1996	8779603
characteristics of transposon insertion mutants of mandelic acid-utilizing pseudomonas putida strain a10l.	a soil isolate, pseudomonas putida strain a10l that utilizes mandelate via the mandelate pathway was mutagenized by transposon tn5-mob insertion and mutant 168 lacking mandelate racemase (mr) and a mutant 254 lacking benzoylformate decarboxylase (bfdc) were obtained. expression of (s)-mandelate dehydrogenase (mdh), bfdc, nad(+)-dependent benzaldehyde dehydrogenase (bdh) and nadp(+)-dependent bdh in the mr-lacking mutant was not affected by the insertion, and it was inducible similarly to the wil ...	1996	8782397
cytochemical colocalization and quantitation of phenotypic and genotypic characteristics in individual bacterial cells.	the widely accepted view that most bacterial species have yet to be cultivated in vitro has gained support from recent ribosomal dna-based environmental studies. to enable elucidation of the phenotypes of organisms recognized solely by molecular genetic techniques, we developed and evaluated cytochemical methods which colocalize phenotypic properties with in situ rrna probe hybridization signals. application of these methods to artificial mixtures of pseudomonas putida and escherichia coli or vi ...	1996	8787385
rapid purification of the oxygenase component of toluene dioxygenase from a polyol-responsive monoclonal antibody.	a monoclonal antibody designated 302 beta that is specific for the beta subunit of the oxygenase component (isptol) of toluene dioxygenase from pseudomonas putida f1 was used to prepare an immunoaffinity column. isptol in cell extracts of escherichia coli jm109(pdtg611) bound to the column, and an enzyme-linked immunosorbent elution-screening assay with different combinations of polyols and kosmotropic anions was used to determine the conditions necessary for recovery of active enzyme. elution f ...	1996	8787410
regio- and stereospecific oxidation of 9,10-dihydroanthracene and 9,10-dihydrophenanthrene by naphthalene dioxygenase: structure and absolute stereochemistry of metabolites.	the oxidation of 9,10-dihydroanthracene and 9,10-dihydrophenanthrene was examined with mutant and recombinant strains expressing naphthalene dioxygenase from pseudomonas putida (ncib 9816.4. salicylate-induced cells of p. putida strain 9816/11 and isopropylthiogalactopyranoside-induced cells of escherichia coli jm109(de3)(pdtg141) oxidized 9,10-dihydroanthracene to (+)-cis-1r,2s)-1,2-dihydroxy-1,2,9,10-tetrahydroanthracene (> 95% relative yield; > 95% enantiomeric excess) as the major product. 9 ...	1996	8795226
increased mutagenesis mediated by cloned plasmid cam-oct genes: potential for expanding substrate ranges of pseudomonas spp.	twenty-five kilobases of pseudomonas plasmid cam-oct dna encoding a dna repair gene(s) was cloned into the broad-host-range vector pvk100. the presence of the cloned genes increased the isolation frequency of pseudomonas putida derivatives capable of using ethyl lactate or 3-methyl-3-buten-1-ol as their carbon source 15- and 8-fold, respectively, after uv irradiation. ethyl lactate-utilizing strains expressed a novel intracellular hydrolase.	1996	8795249
crystallization and preliminary x-ray analysis of salicylate hydroxylase from pseudomonas putida s-1.	apo-salicylate hydroxylase from pseudomonas putida s-1 has been crystallized by the dialysis method, using ammonium sulfate as the precipitant. the crystals belong to hexagonal space group p6(2) or p6(4) with unit cell dimensions of a = b = 142.8 a and c = 63.8 a, and diffract x-rays at higher than 3.5 a resolution. a heavy-atom derivative has been prepared by soaking a crystal in an ammonium sulfate solution containing p-chloromercuriphenylsulfonate.	1996	8797079
cloning, expression, and sequence analysis of the three genes encoding quinoline 2-oxidoreductase, a molybdenum-containing hydroxylase from pseudomonas putida 86.	the three genes coding for quinoline 2-oxidoreductase (qor) of pseudomonas putida 86 were cloned and sequenced. the qor genes are clustered in the transcriptional order medium (m) small (s), large (l) and code for three subunits of 288 (qorm), 168 (qors), and 788 (qorl) amino acids, respectively. formation of active quinoline 2-oxidoreductase and degradation of quinoline occurred in a recombinant p. putida kt2440 clone. the amino acid sequences of qor show significant homology to various prokary ...	1996	8798497
the assimilation of sulfur from multiple sources and its correlation with expression of the sulfate-starvation-induced stimulon in pseudomonas putida s-313.	conditions were optimized for the batch growth of pseudomonas putida s-313 under sulfur-limited conditions. p. putida grew exponentially with sulfate as the sole source of sulfur, and growth was concomitant with the utilization of sulfate until it was exhausted. a further 20% of protein was synthesized after the apparent disappearance of sulfate. a mass balance for the utilized sulfate in cell material was calculated, given the observed molar growth yield of about 3.6 kg protein (mol s)-1 and a ...	1996	8800815
tetralin as a substrate for camphor (cytochrome p450) 5-monooxygenase.	camphor (cytochrome p450) 5-monooxygenase, originally isolated from the bacterium pseudomonas putida pgg 786, catalyzes the essentially stereospecific conversion of tetralin (1,2,3,4-tetrahydronaphthalene) to (r)-1-tetralol ((r).(-)-1,2,3,4-tetrahydro-1-naphthol): tetralin(aq) + nadh(aq) + o2(aq) = (r)-1-tetralol(aq) + nad(aq) + h2o(l). the ratio of the amount of (s)-1-tetralol to the amount of (r)-1-tetralol is small (approximately 0.04) and the reaction is essentially stereospecific. the react ...	1996	8806731
adhesion of the positively charged bacterium stenotrophomonas (xanthomonas) maltophilia 70401 to glass and teflon.	medical implants are often colonized by bacteria which may cause severe infections. the initial step in the colonization, the adhesion of bacteria to the artificial solid surface, is governed mainly by long-range van der waals and electrostatic interactions between the solid surface and the bacterial cell. while van der waals forces are generally attractive, the usually negative charge of bacteria and solid surfaces leads to electrostatic repulsion. we report here on the adhesion of a clinical i ...	1996	8808938
bacterial cytochromes p-450.	the cytochromes p-450 (p-450s) constitute an extremely large family ('superfamily') of haemoproteins that catalyse the oxidation of a wide range of physiological and non-physiological compounds. a remarkable feature of the p-450s is the manipulation of the same basic structure and chemistry to achieve an enormous range of functions in organisms as diverse as bacteria and man. indeed, the p-450s have been described as 'the most versatile biological catalyst known'. much research is focussed on ma ...	1996	8809764
the two-step lysis system of pneumococcal bacteriophage ej-1 is functional in gram-negative bacteria: triggering of the major pneumococcal autolysin in escherichia coli.	the holin function ejh of the pneumococcal bacteriophage ej-1 has been characterized. it shows structural features similar to, and functionally complemented, the prototype member of the holin family. in escherichia coli and pseudomonas putida the ejh product caused cellular death, and changes in cell morphology could be accounted for by lesions in the cytoplasmic membrane. expression of ejh resulted in the inhibition of growth in a variety of phylogenetically distant bacterial genera, suggesting ...	1996	8820638
involvement of sigma 54 in exponential silencing of the pseudomonas putida tol plasmid pu promoter.	the sigma 54-dependent pu promoter of the tol plasmid pww0 of pseudomonas putida becomes activated by the prokaryotic enhancer-binding xyir protein when cells encounter m-xylene in the medium. however, even in the presence of the aromatic inducer, pu activity is silenced in vivo during rapid exponential growth of the cells in rich medium. various elements known to be involved in the control of the transcriptional activity of the promoter were examined to ascertain the mechanism by which expressi ...	1996	8821932
characterization and expression of the plasmid-borne bedd gene from pseudomonas putida ml2, which codes for a nad+-dependent cis-benzene dihydrodiol dehydrogenase.	the catabolic plasmid phmt112 in pseudomonas putida ml2 contains the bed gene cluster encoding benzene dioxygenase (bedc1c2ba) and a nad+-dependent dehydrogenase (bedd) required to convert benzene into catechol. analysis of the nucleotide sequence upstream of the benzene dioxygenase gene cluster (bedc1c2ba) revealed a 1,098-bp open reading frame (bedd) flanked by two 42-bp direct repeats, each containing a 14-bp sequence identical to the inverted repeat of is26. in vitro translation analysis sho ...	1996	8824602
the nucleotide sequence of the pseudomonas aeruginosa pyre-crc-rph region and the purification of the crc gene product.	the gene (crc) responsible for catabolite repression control in pseudomonas aeruginosa has been cloned and sequenced. flanking the crc gene are genes encoding orotate phosphoribosyl transferase (pyre) and rnase ph (rph). new crc mutants were constructed by disruption of the wild-type crc gene. the crc gene encodes an open reading frame of 259 amino acids with homology to the apurinic/apyrimidinic endonuclease family of dna repair enzymes. however, crc mutants do not have a dna repair phenotype, ...	1996	8824606
characterization of an oprl null mutant of pseudomonas putida.	a pseudomonas putida oprl null mutant was generated with reverse genetics by using an in vitro-truncated oprl::xyle construct and in vivo allelic exchange. the nature of the mutation introduced in p. putida was confirmed by southern blotting. western blots (immunoblots) of peptidoglycan-associated proteins revealed that the oprl protein was not made in the mutant strain, whereas it was detectable as a 19-kda band in protein preparations of the wild-type strain. the p. putida oprl, mutant exhibit ...	1996	8824639
role of sigma s in transcription from the positively controlled pm promoter of the tol plasmid of pseudomonas putida.	transcription from the tol plasmid pm promoter is dependent on the xyls regulator activated by benzoate effectors. we analysed transcription from pm in several backgrounds with differing escherichia coli alpha and sigma subunits of rna polymerase. in different rpoa backgrounds, transcription from pm was as high as in the wild-type background throughout the growth curve. in the sigma s-deficient background provided by e. coli rh90, high levels of transcription from pm (xyls/3-methylbenzoate depen ...	1995	8825089
ptim3, a plasmid delivery vector for a transposon-based inducible marker gene system in gram-negative bacteria.	ptim3 is a suicide plasmid vector for the delivery of a transposition defective derivative of tn5, expressing inducible mercury resistance (hgr) and catechol 2,3-dioxygenase (c23o) activity, to a range of gram-negative microorganisms. ptim3 was constructed by a four-stage process from pnmm1, a derivative of psup5011 containing a modified tn5 where antibiotic-resistance determinants have been replaced by the mer operon of tn501 and tdnc (encoding a c23o). in ptim3, tdnc is fused to merd, to bring ...	1995	8825369
active efflux of toluene in a solvent-resistant bacterium.	we investigated the mechanisms behind the organic-solvent resistance of the solvent-tolerant strain pseudomonas putida s12. by use of 14c-labeled toluene, we obtained evidence that an energy-dependent export system may be responsible for this resistance to toluene.	1996	8830706
gram-negative bacteria can induce contact lens related acute red eye (clare) responses.	twelve volunteers participated in a study designed to measure the overnight corneal edema response with a variety of hydrogel contact lenses. during the study four subjects (5 eyes) experienced a contact lens related acute red eye (clare) reaction, which manifested as severe ocular pain, photophobia, corneal infiltration, and conjunctival hyperemia. an additional five subjects (7 eyes) developed corneal infiltrates only. twelve eyes (of 9 subjects) showed no response.	1996	8835069
trans unsaturated fatty acids in bacteria.	the occurrence of trans unsaturated fatty acids as by-products of fatty acid transformations carried out by the obligate anaerobic ruminal microflora has been well known for a long time. in recent years, fatty acids with trans configurations also have been detected in the membrane lipids of various aerobic bacteria. besides several psychrophilic organisms, bacteria-degrading pollutants, such as pseudomonas putida, are able to synthesize these compounds de novo. in contrast to the trans fatty aci ...	1996	8835399
cloning of the genes for and characterization of the early stages of toluene and o-xylene catabolism in pseudomonas stutzeri ox1.	in order to study the toluene and o-xylene catabolic genes of pseudomonas stutzeri ox1, a genomic library was constructed. a 28-kb ecori restriction endonuclease dna fragment, cloned into the vector plasmid plafr1 and designated pfb3401, permitted pseudomonas putida paw340 to convert toluene and o-xylene into the corresponding meta-ring fission products. physical and functional endonuclease restriction maps have been derived from the cloned dna fragment. further subcloning into and deletion anal ...	1996	8837426
amino acid sequence of catechol 1,2-dioxygenase (pyrocatechase) isozyme alpha alpha from pseudomonas arvilla c-1.	catechol 1,2-dioxygenase catalyzes the oxygenative ring cleavage of catechol to form cis,cis-muconic acid, and contains a ferric form of iron as its sole cofactor. pseudomonas arvilla c-1 contains three isozymes of the enzyme, alpha alpha, alpha beta, and beta beta [c. nakai et al. (1990) j. biol. chem. 265, 660-665]. we have determined the amino acid sequence of the alpha alpha isozyme by direct analysis. the sequence shared 77% homology with isozyme beta beta, and had conserved tyrosyl and his ...	1996	8843347
transcriptional regulation of the rhodococcus rhodochrous j1 nita gene encoding a nitrilase.	the 1.4-kb downstream region from a nitrilase gene (nita) of an actinomycete rhodococcus rhodochrous j1, which is industrially in use, was found to be required for the isovaleronitrile-dependent induction of nitrilase synthesis in experiments using a rhodococcus-escherichia coli shuttle vector pk4 in a rhodococcus strain. sequence analysis of the 1.4-kb region revealed the existence of an open reading frame (nitr) of 957 bp, which would encode a protein with a molecular mass of 35,100. deletion ...	1996	8855219
2,4-dioxygenases catalyzing n-heterocyclic-ring cleavage and formation of carbon monoxide. purification and some properties of 1h-3-hydroxy-4-oxoquinaldine 2,4-dioxygenase from arthrobacter sp. rü61a and comparison with 1h-3-hydroxy-4-oxoquinoline 2,4-dioxygenase from pseudomonas putida 33/1.	1h-3-hydroxy-4-oxoquinaldine 2,4-dioxygenase (meqdo) was purified from quinaldine-grown arthrobacter sp. rü61a. it was enriched 59-fold in a yield of 22%, and its properties were compared with 1h-3-hydroxy-4-oxoquinoline 2,4-dioxygenase (qdo) purified from pseudomonas putida 33/1. the enzyme-catalyzed conversions were performed in an (18o)o2/(16o)o2 atmosphere. two oxygen atoms of either (18o)o2 or (16o)o2 were incorporated at c2 and c4 of the respective substrates, indicating that these unusual ...	1996	8856057
crystallization and preliminary x-ray crystallographic studies of ketosteroid isomerase from pseudomonas putida biotype b.	the delta(5)-3-ketosteroid isomerase from pseudomonas putida biotype b has been crystallized. the crystals belong to the space group p2(1)2(1)2(1) with unit cell dimensions of a = 36.48 angstrom, b = 74.30 angstrom, c = 96.02 angstrom, and contain one homodimer per asymmetric unit. native diffraction data to 2.19 angstrom resolution have been obtained from one crystal at room temperature indicating that the crystals are quite suitable for structure determination by multiple isomorphous replaceme ...	1996	8859999
cloning and characterization of the alcaligenes eutrophus 2-oxoglutarate dehydrogenase complex.	nucleotide sequence analysis of a 3.3-kb genomic ecori fragment and of relevant subfragments of a genomic 13.2-kb smai fragment of alcaligenes eutrophus, which were identified by using a dihydrolipoamide dehydrogenase-specific dna probe, revealed the structural genes of the 2-oxoglutarate dehydrogenase complex in a 7.5-kb genomic region. the genes odha (2850 bp), odhb (1248 bp), and odhl (1422 bp), encoding 2-oxoglutarate dehydrogenase (e1), dihydrolipoamide succinyltransferase (e2), and dihydro ...	1996	8867378
cis/trans isomerization of unsaturated fatty acids as possible control mechanism of membrane fluidity in pseudomonas putida p8.	exponentially growing cells of pseudomonas putida had an increased ratio of saturated to unsaturated fatty acids in response to increased growth temperatures. resting cells in which fatty acid biosynthesis was stopped reacted to a thermal increase by converting cis-monounsaturated fatty acids to trans isomers. cis/trans isomerization of up to 60% of the unsaturated fatty acids was also activated by alcohols of different chain length. their effective concentrations apparently depended on the lipo ...	1996	8869883
bacteremia due to glucose non-fermenting gram-negative bacilli in patients with hematological neoplasias and solid tumors.	twenty-six patients with hematological or solid tumors who developed bacteremia caused by stenotrophomonas maltophilia (n = 10), pseudomonas putida (n = 6), sphingomonas paucimobilis complex (n = 4) or alcaligenes xylosoxidans (n = 6) in the period between 1993 and 1995 were studied. seventeen patients were neutropenic during the infection, and 13 were undergoing bone marrow or peripheral blood stem cell transplantation. twenty-three patients had catheter-related infections; only 3 of the 26 pat ...	1996	8874083
accelerated biodegradation of high and low concentrations of p-nitrophenol (pnp) by bacterial inoculation in industrial wastewater: the role of inoculum size on acclimation period	the effect of inoculum size on the acclimation period and rate and extent of p-nitrophenol (pnp) degradation at high (1-10 mg/l) and low (26 &mgr;g/l) concentrations for two bacteria was determined in defined media as well as industrial wastewater. increased inoculum size did not affect the acclimation period of either bacterium at high (1-10 mg/l) pnp concentrations. at low pnp concentrations (26 &mgr;g/l), the two bacteria behaved differently. the acclimation period was shortened and both the ...	1996	8875908
structure and function of the pseudomonas putida integration host factor.	integration host factor (ihf) is a dna-binding and -bending protein that has been found in a number of gram-negative bacteria. here we describe the cloning, sequencing, and functional analysis of the genes coding for the two subunits of ihf from pseudomonas putida. both the ihfa and ihfb genes of p. putida code for 100-amino-acid-residue polypeptides that are 1 and 6 residues longer than the escherichia coli ihf subunits, respectively. the p. putida ihfa and ihfb genes can effectively complement ...	1996	8892836
the beta-ketoadipate pathway and the biology of self-identity.	the beta-ketoadipate pathway is a chromosomally encoded convergent pathway for aromatic compound degradation that is widely distributed in soil bacteria and fungi. one branch converts protocatechuate, derived from phenolic compounds including p-cresol, 4-hydroxybenzoate and numerous lignin monomers, to beta-ketoadipate. the other branch converts catechol, generated from various aromatic hydrocarbons, amino aromatics, and lignin monomers, also to beta-ketoadipate. two additional steps accomplish ...	1996	8905091
electron transfer in zinc-reconstituted nitrite reductase from pseudomonas aeruginosa.	1. the catalytic cycle of the haem-containing nitrite reductase (nir) from pseudomonas aeruginosa involves electron transfer between the two prosthetic groups of the enzyme, the c-haem and the d1-haem; this reaction was shown to be slow by stopped-flow analysis. the recombinant enzyme, expressed in pseudomonas putida, contains the c-haem but no d1-haem; we have reconstituted this protein with zn-protoporphyrin ix in the place of the d1-haem. 2. photoexcitation of zn-nir is followed by electron t ...	1996	8912674
is1394 from pseudomonas alcaligenes n.c.i.b. 9867: identification and characterization of a member of the is30 family of insertion elements.	a new insertion sequence designated is1394 was isolated from pseudomonas alcaligenes ncib 9867 (p25x) by entrapment in plasmid pucd800 which carries the bacillus subtilis sacb and sacr genes. the 1100-bp sequence contains 27-bp inverted repeats with 4 bp mismatch and has one long open reading frame, spanning 92.1% of the entire is. the deduced 338 amino-acid sequence demonstrated homology (varying from 65% to 78% similarity and 36-67% identity) to transposases encoded by the is30 family of is el ...	1996	8917085
oxidation of 6,7-dihydro-5h-benzocycloheptene by bacterial strains expressing naphthalene dioxygenase, biphenyl dioxygenase, and toluene dioxygenase yields homochiral monol or cis-diol enantiomers as major products.	bacterial strains expressing naphthalene, biphenyl, and toluene dioxygenase were examined for their abilities to oxidize 6,7-dihydro-5h-benzocycloheptene (benzocyclohept-1-ene). the major oxidation products were isolated, and their absolute configurations were determined by chiral 1h nuclear magnetic resonance analysis of diastereomeric boronate esters, chiral stationary-phase high-pressure liquid chromatography, and stereo-chemical correlation. pseudomonas sp. strain 9816/11 and sphingomonas ya ...	1996	8919798
expression and substrate specificity of the toluene dioxygenase of pseudomonas putida ncimb 11767.	pseudomonas putida ncimb 11767 oxidized phenol, monochlorophenols, several dichlorophenols and a range of alkylbenzenes (c1-c6) via an inducible toluene dioxygenase enzyme system. biphenyl and naphthalene were also oxidized by this enzyme. growth on toluene and phenol induced the meta-ring-fission enzyme, catechol 2,3-oxygenase, whereas growth on benzoate, which did not require expression of toluene dioxygenase, induced the ortho-ring-cleavage enzyme, catechol 1,2-oxygenase. monochlorobenzoate i ...	1996	8920179
trichloroethylene degradation and mineralization by pseudomonads and methylosinus trichosporium ob3b.	to examine the trichloroethylene (c2hcl3)-degrading capability of five microorganisms, the maximum rate, extent, and degree of c2hcl3 mineralization were evaluated for pseudomonas cepacia g4, pseudomonas cepacia g4 pr1, pseudomonas mendocina kr1, pseudomonas putida f1, and methylosinus trichosporium ob3b using growth conditions commonly reported in the literature for expression of oxygenases responsible for c2hcl3 degradation. by varying the c2hcl3 concentration from 5 microm to 75 microm, vmax ...	1996	8920197
cell-free extract(s) of pseudomonas putida catalyzes the conversion of cyanides, cyanates, thiocyanates, formamide, and cyanide-containing mine waters into ammonia.	our isolate, pseudomonas putida, is known to be capable of utilizing cyanides as the sole source of carbon (c) and nitrogen (n) both in the form of free cells and cells immobilized in calcium alginate. in the present study, the cell-free extract(s) were prepared from the cells of p. putida grown in the presence of sodium cyanide. the ability of enzyme(s) to convert cyanides, cyanates, thiocyanates, formamide and cyanide-containing mine waters into ammonia (nh3) was studied at ph 7.5 and ph 9.5. ...	1996	8920201
biodegradation of phenol by free and immobilized cells of pseudomonas putida.	results of phenol biodegradation by both, free and immobilized pseudomonas putida cells are presented. the influence of ph and the initial substrate concentration was analyzed. on the other hand, several tests of cell adaptation were carried out in order to increase the capacity and the rate of the biodegradation. the behaviour of free and immobilized microorganisms, was similar, reaching a maximum biodegradation capacity of 2000 ppm at an initial ph 6.6 and temperature 30 degrees c. finally, tw ...	1995	8934668
cloning and expression in pseudomonas putida of two of the histidine utilization genes from rhizobium fredii.	two of the genes encoding histidine utilization (hut) in rhizobium fredii strain hh303 have been cloned in pseudomonas putida and partially characterized. molecular cloning of the genes was achieved by mobilizing an r. fredii cosmid library into a mutant strain of p. putida containing a tn5 element in its histidase (huth) gene. a number of overlapping clones were identified, all of which contain a 7.1-kbp hindiii fragment. the origin of this 7.1-kbp fragment from the chromosome of r. fredii was ...	1997	8939803
pcr detection of metallo-beta-lactamase gene (blaimp) in gram-negative rods resistant to broad-spectrum beta-lactams.	we applied pcr to the rapid detection of the metallo-beta-lactamase gene, blaimp, in clinically isolated gram-negative rods. a total of 54 high-level ceftazidime-resistant strains (mics, > 128 micrograms/ml) were subjected to pcr analyses with the blaimp-specific primers, since the blaimp-bearing clinical isolates tested in our previous study always demonstrated high-level resistance to ceftazidime. twenty-two blaimp-positive strains including 9 pseudomonas aeruginosa, 9 serratia marcescens, 2 a ...	1996	8940421
the amino acid sequence of rat kidney 5-oxo-l-prolinase determined by cdna cloning.	5-oxoprolinase (ec 3.5.2) catalyzes a reaction in which the endergonic cleavage of 5-oxo-l-proline to form l-glutamate is coupled to the exergonic hydrolysis of atp to adp and inorganic phosphate. highly purified preparations of the enzyme have been obtained from rat kidney and pseudomonas putida. the rat kidney enzyme is composed of two strongly interacting, apparently identical subunits (mr = 142,000), whereas that from p. putida is composed of two functionally different protein components tha ...	1996	8943290
analysis of cumene (isopropylbenzene) degradation genes from pseudomonas fluorescens ip01.	we obtained the dna fragments encoding 2-hydroxy-6-oxo-7-methylocta-2,4-dienoic acid (homoda) hydrolase in the cumene (isopropylbenzene) degrader pseudomonas fluorescens strain ip01 via pcr using two synthesized oligonucleotides corresponding to the conserved regions within known meta-cleavage compound hydrolases. following colony hybridization using the amplified dna as a probe, a 4.5-kb hindiii fragment was isolated from p. fluorescens ip01. after determining the nucleotide sequence of this fr ...	1996	8953719
activity and three-dimensional distribution of toluene-degrading pseudomonas putida in a multispecies biofilm assessed by quantitative in situ hybridization and scanning confocal laser microscopy.	as a representative member of the toluene-degrading population in a biofilter for waste gas treatment, pseudomonas putida was investigated with a 16s rrna targeting probe. the three-dimensional distribution of p. putida was visualized in the biofilm matrix by scanning confocal laser microscopy, demonstrating that p. putida was present throughout the biofilm. acridine orange staining revealed a very heterogeneous structure of the fully hydrated biofilm, with cell-free channels extending from the ...	1996	8953734
characterization of a 7fe ferredoxin isolated from the marine denitrifier pseudomonas nautica strain 617: spectroscopic and electrochemical studies.	a 7fe ferredoxin, isolated from the marine denitrifier pseudomonas nautica strain 617, was characterized. the nh2-terminal sequence analysis, performed until residue number 56, shows a high similarity with the 7fe ferredoxins isolated from azotobacter vinelandii, pseudomonas putida, and pseudomonas stutzeri. epr and nmr spectroscopies identify the presence of both [3fe-4s] and [4fe-4s] clusters, with cysteinyl coordination. the electrochemical studies on [fe-s] clusters show that a fast diffusio ...	1996	8954931
mannitol, a novel bacterial compatible solute in pseudomonas putida s12.	the aim of this study was to identify the compatible solutes accumulated by pseudomonas putida s12 subjected to osmotic stress. in response to reduced water activity, p. putida s12 accumulated nalpha-acetylglutaminylglutamine amide (naggn) simultaneously with a novel compatible solute identified as mannitol (using 13c- and 1h-nuclear magnetic resonance, liquid chromatography-mass spectroscopy and high-performance liquid chromatography methods) to maximum concentrations of 74 and 258 micromol g ( ...	1996	8955280
identification and characterization of the tolqra genes of pseudomonas aeruginosa.	the tolq, tolr, and tola genes from pseudomonas aeruginosa pao were cloned using degenerate oligonucleotide pcr primers designed based on conserved transmembrane regions of escherichia coli tolq and tolr and e. coli and pseudomonas putida exbb and exbd. the resulting pcr product was used as a probe to isolate a 6.5-kb dna fragment containing p. aeruginosa tolq, tolr, and tola. the nucleotide sequence of a 2.9-kb dna fragment containing the tolq, tolr, and tola genes was determined. the dna seque ...	1996	8955385
clinical spectrum of pseudomonas putida infection.	the clinical and microbiologic characteristics of 55 cases of pseudomonas putida infection in 53 patients in a medical center in taiwan from april 1988 to march 1993 are reported. p. putida was cultured in the decreasing order of frequency from urine (24 isolates), sputum (12), blood (10), wound discharge (3), peritoneal fluid (3), cerebrospinal fluid (2) and umbilical swab (1). of the adult patients, 23% (12/53) were considered to be contaminated or colonized with p. putida. of the 41 patients ...	1996	8961672
the effect of nutrient limitation on styrene metabolism in pseudomonas putida ca-3.	styrene degradation in pseudomonas putida ca-3 has previously been shown to be subject to catabolite repression in batch culture. we report here on the catabolite-repressing effects of succinate and glutamate and the effects of a limiting inorganic-nutrient concentration on the styrene degradation pathway of p. putida ca-3 in a chemostat culture at low growth rates (0.05 h-1). oxidation of styrene and the presence of styrene oxide isomerase and phenylacetaldehyde dehydrogenase activities were us ...	1996	8967774
molecular cloning and expression in different microbes of the dna encoding pseudomonas putida u phenylacetyl-coa ligase. use of this gene to improve the rate of benzylpenicillin biosynthesis in penicillium chrysogenum.	the gene encoding phenylacetyl-coa ligase (pcl), the first enzyme of the pathway involved in the aerobic catabolism of phenylacetic acid in pseudomonas putida u, has been cloned, sequenced, and expressed in two different microbes. in both, the primary structure of the protein was studied, and after genetic manipulation, different recombinant proteins were analyzed. the pcl gene, which was isolated from p. putida u by mutagenesis with the transposon tn5, encodes a 48-kda protein corresponding to ...	1996	8969218
comparison of factors influencing trichloroethylene degradation by toluene-oxidizing bacteria.	the degradation of trichloroethylene (tce) by toluene-oxidizing bacteria has been extensively studied, and yet the influence of environmental conditions and physiological characteristics of individual strains has received little attention. to consider these effects, the levels of tce degradation by strains distinguishable on the basis of toluene and nitrate metabolism were compared under aerobic or hypoxic conditions in the presence and absence of nitrate and an exogenous electron donor, lactate ...	1996	8975612
succession and convergence of biofilm communities in fixed-film reactors treating aromatic hydrocarbons in groundwater.	community composition, succession, and performance were compared in three fluidized bed reactors (fbr) operated to test preemptive colonization and the influence of toluene compared with a mixture of benzene, toluene, and p-xylene (btx) as feeds. one reactor was inoculated with toluene-degrading strains pseudomonas putida paw1, burkholderia cepacia g4, and b. pickettii pko1. paw1 outcompeted the other two strains. when groundwater strains were allowed to challenge the steady-state biofilm develo ...	1997	8979355