Publications

cloning and sequencing of a 29 kb region of the bacillus subtilis genome containing the hut and wapa loci.	within the framework of an international project for the sequencing of the entire bacillus subtilis genome, a 29 kb chromosome segment, which contains the hut operon (335 degrees) and the wapa gene, has been cloned and sequenced. this region (28,954 bp) contains 21 complete orfs and one partial one. the 5th, 6th and 17th genes correspond to huth encoding histidase, hutp encoding the positive regulator for the hut operon and wapa encoding a precursor of three major wall-associated proteins, respe ...	1995	7704263
[degradation of syntanol ds-10 by bacteria immobilized in polysaccharide gels].	degradative capability of bacterium pseudomonas putida immobilized in polysaccharide (agar-agar or furcellarane) gels towards a nonionic surfactant sintanol ds-10 was studied. the immobilized cells were shown to retain their ability to degrade the substrate in the model experiments as well as in the real waste water. the substrate was degraded after adsorption to the carrier beads.	1995	7746826
overlapping substrate specificities of benzaldehyde dehydrogenase (the xylc gene product) and 2-hydroxymuconic semialdehyde dehydrogenase (the xylg gene product) encoded by tol plasmid pww0 of pseudomonas putida.	two aldehyde dehydrogenases involved in the degradation of toluene and xylenes, namely, benzaldehyde dehydrogenase and 2-hydroxymuconic semialdehyde dehydrogenase, are encoded by the xylc and xylg genes, respectively, on tol plasmid pww0 of pseudomonas putida. the nucleotide sequence of xylc was determined in this study. a protein exhibiting benzaldehyde dehydrogenase activity had been purified from cells of p. putida (pww0) (j. p. shaw and s. harayama, eur. j. biochem. 191:705-714, 1990); howev ...	1995	7868591
a manganese-dependent dioxygenase from arthrobacter globiformis cm-2 belongs to the major extradiol dioxygenase family.	almost all bacterial ring cleavage dioxygenases contain iron as the catalytic metal center. we report here the first available sequence for a manganese-dependent 3,4-dihydroxyphenylacetate (3,4-dhpa) 2,3-dioxygenase and its further characterization. this manganese-dependent extradiol dioxygenase from arthrobacter globiformis cm-2, unlike iron-dependent extradiol dioxygenases, is not inactivated by hydrogen peroxide. also, ferrous ions, which activate iron extradiol dioxygenases, inhibit 3,4-dhpa ...	1995	7868595
lipase modulator protein (liml) of pseudomonas sp. strain 109.	plasmids containing a pseudomonas sp. strain 109 extracellular lipase gene (lipl) lacking nh2-terminal sequence and a lipase modulator gene (liml) lacking the nh2-terminal hydrophobic region were constructed and expressed independently in escherichia coli by using the t7 promoter expression vector system. recombinant lipl (rlipl) was produced as inclusion bodies, whereas recombinant liml (rliml) was present as a soluble protein. during in vitro renaturation of the purified rlipl inclusion bodies ...	1995	7868599
discontinuities in the evolution of pseudomonas putida cat genes.	the organization and transcriptional control of chromosomal cat genes (required for dissimilation of catechol by the beta-ketoadipate pathway) in the pseudomonas putida biotype strain (atcc 12633) are reported. nucleotide sequence reveals that catr is separated by 135 bp from the divergently transcribed catbc,a; catc begins 21 nucleotides downstream from catb, and cata begins 41 nucleotides downstream from catc. this contrasts with the gene arrangement in other bacteria, in which cata lies sever ...	1995	7814330
regulation of the hema gene during 5-aminolevulinic acid formation in pseudomonas aeruginosa.	the general tetrapyrrole precursor 5-aminolevulinic acid is formed in bacteria via two different biosynthetic pathways. members of the alpha group of the proteobacteria use 5-aminolevulinic acid synthase for the condensation of succinyl-coenzyme a and glycine, while other bacteria utilize a two-step pathway from aminoacylated trna(glu). the trna-dependent pathway, involving the enzymes glutamyl-trna reductase (encoded by hema) and glutamate-1-semialdehyde-2,1-aminomutase (encoded by heml), was d ...	1995	7883699
aromatic effector activation of the ntrc-like transcriptional regulator phhr limits the catabolic potential of the (methyl)phenol degradative pathway it controls.	pseudomonas putida p35x (ncib 9869) metabolizes phenol and monomethylphenols via a chromosomally encoded meta-cleavage pathway. we have recently described a 13.4-kb fragment of the chromosome that codes for the first eight genes of the catabolic pathway and a divergently transcribed positive regulator, phhr. the eight structural genes lie in an operon, the phh operon, downstream of a -24 tggc, -12 ttgc promoter sequence. promoters of this class are recognized by rna polymerase that utilizes the ...	1995	7883704
single amino acids changes in the signal receptor domain of xylr resulted in mutants that stimulate transcription in the absence of effectors.	the xylr protein positively controls expression from the pseudomonas putida tol plasmid sigma 54-dependent "upper" pathway operon promoter (pu) and the xyls gene promoter (ps), in response to the presence of aromatic effectors. two mutant xylr regulators able to stimulate transcription from pu and ps in the absence of effectors were isolated. these mutants exhibited single point mutations, namely asp135-->asn and pro85-->ser. both mutations are located in the amino termini domain of xylr, which ...	1995	7890623
identification of functional residues in a 2-hydroxymuconic semialdehyde hydrolase. a new member of the alpha/beta hydrolase-fold family of enzymes which cleaves carbon-carbon bonds.	the 2-hydroxymuconic semialdehyde hydrolase, xylf, of the pseudomonas putida tol plasmid-encoded pathway for the catabolism of toluene and xylenes, catalyzes one of the rarest types of enzyme reaction (ec 3.7.1.9), the hydrolysis of a carbon-carbon bond in its substrate, the ring-fission product of 3-alkyl-substituted catechols. in this study, amino acid sequence comparisons between xylf and other hydrolases, and analysis of the similarity between the predicted secondary structure of xylf and th ...	1995	7890778
mechanism of the reaction catalyzed by mandelate racemase: structure and mechanistic properties of the k166r 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 that either abstract the alpha-proton from the enantiomers of mandelate to form an enolic intermediate or protonate the enolic ...	1995	7893690
aspartate transcarbamoylase genes of pseudomonas putida: requirement for an inactive dihydroorotase for assembly into the dodecameric holoenzyme.	the nucleotide sequences of the genes encoding the enzyme aspartate transcarbamoylase (atcase) from pseudomonas putida have been determined. our results confirm that the p. putida atcase is a dodecameric protein composed of two types of polypeptide chains translated coordinately from overlapping genes. the p. putida atcase does not possess dissociable regulatory and catalytic functions but instead apparently contains the regulatory nucleotide binding site within a unique n-terminal extension of ...	1995	7896697
a carbon starvation survival gene of pseudomonas putida is regulated by sigma 54.	by using mini-tn5 transposon mutagenesis, two mutants of pseudomonas putida atcc 12633 were isolated which showed a marked increase in their sensitivity to carbon starvation; these mutants are presumably affected in the pex type of proteins that p. putida induces upon carbon starvation (m. givskov, l. eberl, and s. molin, j. bacteriol. 176:4816-4824, 1994). the affected genes in our mutants were induced about threefold upon carbon starvation. the promoter region of the starvation gene in the mut ...	1995	7896711
pcr amplification and direct sequencing of gyrb genes with universal primers and their application to the detection and taxonomic analysis of pseudomonas putida strains.	degenerate pcr primers, up-1 and up-2r, for the amplification of dna gyrase subunit b genes (gyrb) were designed by using consensus amino acid sequences of gyrases from escherichia coli, pseudomonas putida, and bacillus subtilis. in addition to the degenerate sequences, these primers have sequences at the 5' end which allow direct sequencing of amplified pcr products. with these primers, dna segments of the predicted size were amplified from a variety of gram-negative and gram-positive genera. t ...	1995	7793912
characterization of biphenyl catabolic genes of gram-positive polychlorinated biphenyl degrader rhodococcus sp. strain rha1.	rhodococcus sp. strain rha1 is a gram-positive polychlorinated biphenyl (pcb) degrader which can degrade 10 ppm of pcb48 (equivalent to aroclor1248), including tri-, tetra-, and pentachlorobiphenyls, in a few days. we isolated the 7.6-kb ecori-bamhi fragment carrying the biphenyl catabolic genes of rha1 and determined their nucleotide sequence. on the basis of deduced amino acid sequence homology, we identified six bph genes, bpha1a2a3a4, bphb, and bphc, that are responsible for the initial thre ...	1995	7793929
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
biological production of semisynthetic opiates using genetically engineered bacteria.	semisynthetic derivatives of morphine and related alkaloids are in widespread clinical use. due to the complexity of these molecules, however, chemical transformations are difficult to achieve in high yields. we recently identified the powerful analgesic hydromorphone as an intermediate in the metabolism of morphine by pseudomonas putida m10. here we describe the construction of recombinant strains of escherichia coli that express morphine dehydrogenase and morphinone reductase. these strains ar ...	1995	9634804
amplification of toluene dioxygenase genes in a hybrid pseudomonas strain to enhance the biodegradation of benzene, toluene, and p-xylene mixture.	a hybrid metabolic pathway through which benzene, toluene, and p-xylene (btx) mixture could be simultaneously mineralized was previously constructed in pseudomonas putida tb101 (lee, roh, kim, biotechnol. bioeng 43: 1146-1152, 1994). in this work, we improved the performance of the hybrid pathway by cloning the todc1c2ba genes in the broad-host-range multicopy vector rsf1010 and by introducing the resulting plasmid ptol037 into p. putida mt-2 which harbors the archetypal tol plasmid. as a result ...	1995	18623248
modeling simultaneous hexavalent chromium reduction and phenol degradation by a defined coculture of bacteria.	based on the kinetics of cr(vi) reduction by escherichia coli atcc 33456 and phenol degradation by pseudomonas putida dmp-1, a mathematical model is developed to describe simultaneous cr(vi) reduction and phenol degradation in the coculture of the two species. the developed model incorporates the toxicity effects of cr(vi) and phenol on phenol degradation and cr(vi) reduction in the coculture. the model illustrates the inhibitory effects of phenol on cr(vi) reduction and cr(vi) toxicity toward p ...	1995	18623529
biodegradation of nitrobenzene through a hybrid pathway in pseudomonas putida.	the biodegradation of nitrobenzene was attempted by using pseudomonas putida tb 103 which possesses the hybrid pathway combining the tod and the tol pathways. analysis of the metabolic flux of nitrobenzene through the hybrid pathway indicated that nitrobenzene was initially oxidized to cis-1,2-dihydroxy-3-nitrocyclohexa-3,5-diene by toluene dioxygenase in the tod pathway and then channeled into the tol pathway, leading to the complete biodegradation of nitrobenzene. a crucial metabolic step redi ...	1995	18623531
determination of cadmium in biological and environmental samples by slurry electrothermal atomic absorption spectrometry.	the retention of cadmium by the bacteria escherichia coli and pseudomonas putida was optimized in order to develop a rapid and selective preconcentration method for cadmium from biological and environmental samples prior to determination by electrothermal atomic absorption spectrometry. living and lyophilized cells for both bacteria were used, but the method using dead cells shows better analytical capabilities. cadmium from aqueous solutions is easily retained on the outer membrane of both bact ...	1995	18966409
interesterification of butter fat by partially purified extracellular lipases from pseudomonas putida, aspergillus niger and rhizopus oryzae.	three extracellular lipases were produced by batch fermentation of pseudomonas putida atcc 795, aspergillus niger cbs 131.52 and rhizopus oryzae atcc 34612 during the late phase of growth, at 72, 96 and 96 h, respectively. the lipases were partially purified by (nh4)2so4 fractionation. the lipase of p. putida was optimal at ph 8.0 whereas those from a. niger and r. oryzae were optimal at ph 7.5. the a. niger lipase had the lowest v max value (0.51×10(-3) u/min) and r. oryzae the highest (1.86×10 ...	1995	24415019
macro-kinetic investigation on phenol uptake from air by biofiltration: influence of superficial gas flow rate and inlet pollutant concentration.	the macro-kinetic behavior of phenol removal from a synthetic exhaust gas was investigated theoretically as well as experimentally by means of two identical continuously operating laboratory-scale biological filter bed columns. a mixture of peat and glass beads was used as filter material. after sterilization it was inoculated with a pure strain of pseudomonas putida, as employed in previous experimental studies. to determine the influence of the superficial gas flow rate on biofilter performanc ...	1996	18623593
optimization of trichloroethylene degradation using soluble methane monooxygenase of methylosinus trichosporium ob3b expressed in recombinant bacteria.	by complementing cell-free extracts of pseudomonas putida f1/psmmo20 with purified soluble methane monooxygenase (smmo) components of methylosinus trichosporium ob3b, the low cloned-gene smmo activity in the recombinant strain was found to be due to incomplete activity of the hydroxylase component. to address this incomplete activity, additional smmo-expressing strains were formed by transferring mmo-containing psmmo20 and psmmo50 into various bacterial species including pseudomonads and alpha-2 ...	1996	18624367
biodegradation of phenol using the self-cycling fermentation (scf) process.	self-cycling fermentation (scf) in a stirred tank reactor was applied to the biodegradation of phenol by pseudomonas putida. the technique resulted in stable and repeatable performance. complete substrate consumption was achieved under all operating conditions investigated. scf resulted in substrate utilization rates as high as 14.5 kg of phenol per cubic meter of fermentor volume per day of fermentation, higher than those that have been reported for batch, cstr, and packed column fermentors. a ...	1996	18627094
biosynthesis of synthons in two-liquid-phase media.	the pseudomonas oleovorans alkane hydroxylase and xylene oxygenase from pseudomonas putida are versatile mono-oxygenases for stereo- and regioselective oxidation of aliphatic and aromatic hydrocarbons. pseudomonas oleovorans and alkanol dehydrogenase deficient mutants of pseudomonas have previously been used to produce alkanols from various alkanes and optically active epoxides from alkenes. similarly, p. putida strains have been used to produce aromatic alcohols, aromatic acids, and optically a ...	1996	18629897
responses of intracellular cofactors to single and dual substrate limitations.	the highly systematic responses of cellular cofactors to controlled substrate limitations of electron donor, electron acceptor, and both (dual limitation) were quantified using continuous-flow cultures of pseudomonas putida. the results showed that the nadh concentration in the cells decreased gradually as the specific rate of electron-donor utilization (-q(d)) fell or increased systematically as oxygen limitation became more severe for fixed -q(d), while the nad concentration was invariant. the ...	1996	18626865
kinetics of phenol biodegradation in the presence of glucose.	the kinetics of utilization of glucose, phenol, and their mixtures by pseudomonas putida (atcc 17514) were studied with a continuously aerated, jacketed batch reactor operating at 28 degrees c and ph 7.2. it was found that when glucose is the sole carbon and energy source, the culture utilizes it following monod kinetics. when phenol is the sole carbon and energy source, the culture biodegrades it following andrews (inhibitory) kinetics. when both glucose and phenol are present in the medium, th ...	1996	18627091
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
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
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
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
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
organisation of the tmb catabolic operons of pseudomonas putida tmb and evolutionary relationship with the xyl operons of the tol plasmid pww0.	in pseudomonas putida (pp) tmb the genes involved in the catabolism of methyl-substituted aromatic hydrocarbons 1,2,4-trimethylbenzene, m- and p-xylene (tmb operon), are functionally and genetically homologous to the xyl genes of the plasmid pww0, but are chromosomally encoded. we have analysed by cloning. southern blotting and sequencing of selected regions the organisation of the tmb cluster. this analysis shows that the structural and regulatory genes of the tmb and xyl systems exhibit a high ...	1996	8982087
cotranscription of a gtpase gene from the cyanobacterium synechocystis pcc 6803 and a p-type ca(2+)-atpase gene.	a gtpase gene adjacent to the ca(2+)-atpase gene from synechocystis pcc 6803 has been sequenced. it encodes for a protein of 456 amino acids revealing high homology to so-called 50k proteins of bacillus subtilis and pseudomonas putida. cotranscription of gtpase and ca(2+)-atpase genes has been shown by reverse transcription pcr.	1996	8982253
biodegradation of hydrogen sulfide by a laboratory-scale immobilized pseudomonas putida ch11 biofilter.	a heterotrophic pseudomonas putida ch11 was isolated from livestock farming wastewater and applied for the treatment of h2s-containing gas. extensive tests including removal characteristics, metabolic products, and removal efficiencies of h2s by p. putida ch11 were examined in batch and continuous systems. the optimum ph required to remove hydrogen sulfide was found in the range of 6-8. the maximum removal rate and the saturation constant were calculated to be vm = 1.36 g s/day.kg dry bead and k ...	1996	8983205
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
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
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
3-hydroxyisobutyrate dehydrogenase from pseudomonas putida e23: purification and characterization.	the nad(+)-dependent 3-hydroxyisobutyrate dehydrogenase [ec 1.1.1.31] was purified to homogeneity from pseudomonas putida e23. the enzyme was a tetramer (molecular mass, 120 kda) consisted of identical subunits (molecular mass, 30 kda). the enzyme was specific for nad+ (km, 0.44 mm). the maximal activity was obtained at about ph 10. the enzyme was specific for the l-isomer of 3-hydroxyisobutyrate. in addition to l-3-hydroxyisobutyrate, l-serine, 2-methyl-dl-serine, and 3-hydroxypropionate were s ...	1996	8988636
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
cloning and nucleotide sequence of a d,l-haloalkanoic acid dehalogenase encoding gene from alcaligenes xylosoxidans ssp. denitrificans abiv.	we have cloned dna fragments of plasmid pfl40 from alcaligenes xylosoxidans ssp. denitrificans abiv encoding a d,l-2-haloalkanoic acid halidohydrolase (dhliv). a 6.5-kb ecori/sali-fragment with inducible expression of the halidohydrolase was cloned in pseudomonas fluorescens and escherichia coli. a 1.9-kb hindii-fragment demonstrated expression of the dehalogenase only due to the presence of the promoter from the puc vector in escherichia coli. the nucleotide sequence of this dna-fragment was de ...	1996	9144969