Publications

evidence for autoregulation of camr, which encodes a repressor for the cytochrome p-450cam hydroxylase operon on the pseudomonas putida cam plasmid.	the regulatory gene camr on the cam plasmid of pseudomonas putida (atcc 17453) negatively controls expression of the cytochrome p-450cam hydroxylase operon (camdcab) for the camphor degradation pathway and is oriented in a direction opposite to that of the camdcab operon. in this study, we examined expression of the camr gene by monitoring the beta-galactosidase activity of camr-lacz translational fusions in p. putida camr and camr+ strains. we found that the camr gene was autogenously regulated ...	1993	8253671
depletion of serum methionine by methioninase in mice.	methionine dependence is a tumor-specific metabolic defect found in human cancer cell lines as well as in fresh human tumor specimens. methionine dependent tumors cease growing when deprived of methionine, unlike normal cells which can substitute homocysteine for methionine for their growth requirement. we have previously purified a stable, endotoxin-free methioninase from the bacterium, pseudomonas putida. we demonstrate in this report that purified methioninase can lower the serum levels of me ...	1993	8239522
new derivatives of tol plasmid pww0.	two new segregants, ppw1-1 and ppw161-1, of pseudomonas putida were isolated from the stock cultures paw85(pww0) and paw85(pww0-161). strain ppw1-1 had lost its ability to grow on m-xylene but was able to grow on m-toluate. a deletion of the left-hand of transposon tn4651, including the upper-operon genes, had taken place in plasmid pww0mut1, isolated from strain ppw1-1. additional deletions were observed in pww0mut1 after 'benzoate-curing' (plasmids pww0mut15, pww0mut19, pww0mut27). the genes o ...	1993	8254307
a novel structural basis for membrane association of a protein: construction of a chimeric soluble mutant of (s)-mandelate dehydrogenase from pseudomonas putida.	the (s)-mandelate dehydrogenase (mdh) from pseudomonas putida (atcc 12633) is the only membrane-associated member of a homologous family of fmn-dependent, alpha-hydroxy acid dehydrogenases/oxidases that includes the structurally characterized glycolate oxidase from spinach (gox). we have correlated the membrane association of mdh to a polypeptide segment in the interior of the primary sequence. this has been accomplished by construction of a chimeric enzyme in which the putative membrane-binding ...	1993	8241149
regulation of the catechol 1,2-dioxygenase- and phenol monooxygenase-encoding pheba operon in pseudomonas putida paw85.	in pseudomonas putida paw85, the ortho-cleavage pathway is used for catechol degradation. the 11.4-kb xhoi fragment cloned from phenol degradation plasmid pest1226 into pkt240 (recombinant plasmid pat1140) contains the inducible pheba operon that encodes catechol 1,2-dioxygenase (gene pheb) and phenol monooxygenase (gene phea), the first two enzymes for the phenol degradation pathway. the promoter of the pheba operon is mapped 1.5 kb upstream of the pheb gene. the plasmid pat1140, when introduce ...	1993	8253692
use of 5-cyano-2,3-ditolyl tetrazolium chloride for quantifying planktonic and sessile respiring bacteria in drinking water.	direct microscopic quantification of respiring (i.e., viable) bacteria was performed for drinking water samples and biofilms grown on different opaque substrata. water samples or biofilms developed in flowing drinking water were incubated with the vital redox dye 5-cyano-2,3-ditolyl tetrazolium chloride (ctc) and r2a medium. one hour of incubation with 0.5 mm ctc was sufficient to obtain intracellular reduction of ctc to the insoluble fluorescent formazan (ctf) product, which was indicative of c ...	1993	8285688
the preparation of endotoxin-free l-methionine-alpha-deamino-gamma-mercaptomethane-lyase (l-methioninase) from pseudomonas putida.	many types of human and animal tumors have an absolute requirement for methionine. this requirement can be satisfied by homocysteine only in normal cells and tissues. therefore, methionine may be an important target in cancer therapy. to attack this target we have purified endotoxin-free methioninase from pseudomonas putida by a novel and simple procedure. this procedure involves (1) a heat step of the cell extract at 60 degrees c for 8 min, (2) deae-toyopearl ion-exchange chromatography, (3) de ...	1993	8286949
[the role of pyrimidines in the biosynthesis of the fluorescing pigment pyoverdin pm in pseudomonas putida m].	dihydroorotate was shown to be a predecessor of deoxyquinoline nucleus of a fluorescing enzyme pyoverdin pm in pseudomonas putida m. the data was obtained in experiments with a set of pyr- mutants with different steps of pyrimidine synthesis blocked. a scheme for deoxyquinoline nucleus of the enzyme including dihydroorotate participation is proposed.	1993	8289842
microbial metabolism of quinoline and related compounds. xx. quinaldic acid 4-oxidoreductase from pseudomonas sp. ak-2 compared to other procaryotic molybdenum-containing hydroxylases.	quinaldic acid 4-oxidoreductase from pseudomonas sp. ak-2 catalyses the hydroxylation of quinoline 2-carboxylic (quinaldic acid) to 4-hydroxyquinoline 2-carboxylic acid (kynurenic acid) with concomitant reduction of a suitable electron acceptor. an analogous hydroxylation in para-position relative to the n-heteroatom was only recently described for quinaldine 4-oxidoreductase (de beyer & lingens, 1993, biol. chem. hoppe-seyler 374, 101-110) and for quinaldic acid 4-oxidoreductase from serratia m ...	1993	8292263
nucleotide sequence and over-expression of morphine dehydrogenase, a plasmid-encoded gene from pseudomonas putida m10.	pseudomonas putida m10 was originally isolated from factory waste liquors by selection for growth on morphine. the nadp(+)-dependent morphine dehydrogenase that initiates morphine catabolism is encoded by a large plasmid of 165 kb. treatment of p. putida m10 with ethidium bromide led to the isolation of a putative plasmid-free strain that was incapable of growth on morphine. the structural gene for morphine dehydrogenase, mora, has been located on the plasmid by oligonucleotide hybridization, by ...	1993	8452544
increased expression of the plasmid-determined 2,3-dihydroxybiphenyl dioxygenase gene in strains of escherichia coli, pseudomonas putida and pseudomonas aeruginosa.	a 6.5-kb ecori fragment containing the gene encoding 2,3-dihydroxybiphenyl dioxygenase from the plasmid pbs312 was cloned into broad host range plasmid rsf1010 and expressed in escherichia coli, pseudomonas putida and pseudomonas aeruginosa strains. the increased expression of the gene was orientation-dependent and probably due to the transcription read through from the streptomycin promoter of the vector. subcloning experiments of the psti fragments of pbs312 plasmid using vector pbr322 reveale ...	1993	8454186
copper accumulation by a strain of pseudomonas putida.	a study on the copper accumulation by resting cells of copper-resistant bacteria, isolated from activated sludge and electroplating effluent, was conducted. the best selected strain, identified as pseudomonas putida ii-11, retained copper ions, cu(ii), as high as 6.5% of its dry weight. bacterial cells grown in the sulphate-limiting medium had the highest copper removal capacity [rc, mg of cu(ii)/g of dry cells], while the presence of glucose or sodium azide did not affect cu(ii) rc of the bacte ...	1993	8459779
purification of pseudomonas putida acyl coenzyme a ligase active with a range of aliphatic and aromatic substrates.	acyl coenzyme a (acyl-coa) ligase (acyl-coa synthetase [acoas]) from pseudomonas putida u was purified to homogeneity (252-fold) after this bacterium was grown in a chemically defined medium containing octanoic acid as the sole carbon source. the enzyme, which has a mass of 67 kda, showed maximal activity at 40 degrees c in 10 mm k2po4h-napo4h2 buffer (ph 7.0) containing 20% (wt/vol) glycerol. under these conditions, acoas showed hyperbolic behavior against acetate, coa, and atp; the kms calcula ...	1993	8476289
a mutagenesis system utilizing a tn1722 derivative containing an escherichia coli-specific vector plasmid: application to pseudomonas species.	a novel transposon (tn) mutagenesis system for gram- non-enteric bacteria was developed which allowed rapid and one-step cloning of the mutated region in escherichia coli. the tn constructed was tn1722-299km, a tn1722 derivative containing a kmr gene and the entire sequence of an e. coli-specific plasmid, pacyc184. the hybrid plasmid consisting of tn1722-299km and the transfer genes of plasmid r388 was conjugally transferred from e. coli to pseudomonas putida or p. aeruginosa, and selection of t ...	1993	8294012
identification of a cis-acting sequence within the pm promoter of the tol plasmid which confers xyls-mediated responsiveness to substituted benzoates.	the dna sequences within the pm promoter/operator region of the meta operon of the tol plasmid of pseudomonas putida, which confer xyls-mediated responsiveness to substituted benzoates, have been identified through deletion analysis and mutagenesis of the region. integrity and proper phasing of two homologous tandem sequences 5'-tgcaapuaapu-pyggnta-3', separated by six base-pairs and overlapping with the -35 region of the pm promoter, was essential for m-toluate activation of a pm-lacz fusion in ...	1993	8478926
construction of chromosomal reca mutants of pseudomonas putida ppg2.	the reca gene of pseudomonas putida ppg2 was cloned by complementation of the reca mutations of escherichia coli strains dh5 alpha and hb101. the nucleotide sequence of the dna fragment was determined and shown to contain reca and a downstream partial open reading frame. two mutants of p. putida ppg2, strains js387 and js388, were constructed by insertional inactivation of reca with a tetracycline-resistance gene in both orientations. both mutants acquired sensitivity to methyl methanesulfonate ...	1993	8294013
analysis of the dna damage-mediated induction of pseudomonas putida and pseudomonas aeruginosa lexa genes.	a fusion between the lexa gene of pseudomonas aeruginosa and pseudomonas putida and the lacz gene was constructed in vitro and cloned in a mini-tn5 transposon derivative to obtain chromosomal insertions which enable to quantitatively examine their transcriptional regulation in both pseudomonas and e. coli. analysis of dna damage-mediated induction of these lexa-lacz fusions showed that expression of p. putida and p. aeruginosa lexa genes was always higher and earlier than the expression of the l ...	1993	8319897
the bkdr gene of pseudomonas putida is required for expression of the bkd operon and encodes a protein related to lrp of escherichia coli.	branched-chain keto acid dehydrogenase is a multienzyme complex which is required for the metabolism of the branched-chain amino acids in pseudomonas putida. the structural genes encoding all four proteins of the bkd operon have been cloned, and their nucleotide sequences have been determined (g. burns, k. t. madhusudhan, k. hatter, and j. r. sokatch, p. 177-184 in s. silver, a. m. chakrabarty, b. iglewski, and s. kaplan [ed.], pseudomonas: biotransformations, pathogenesis, and evolving biotechn ...	1993	8320210
a new amino acid racemase with threonine alpha-epimerase activity from pseudomonas putida: purification and characterization.	we have found that pseudomonas putida atcc 17642 cells grown in a medium containing d-threonine as the sole nitrogen source produce an enzyme that catalyzes epimerization of threonine. proton nuclear magnetic resonance analysis of the enzyme reaction in deuterium oxide clearly showed epimerization from l- to d-allo-threonine and also from d- to l-allo-threonine. this is the first example of an enzyme that was clearly shown to epimerize threonine. the enzyme has been purified to homogeneity, whic ...	1993	8320235
degradative capability of pseudomonas putida on acetonitrile.	pseudomonas putida, capable of utilizing acetonitrile as a sole source of carbon and nitrogen, was isolated from contaminated soil and water samples collected from industrial sites. the p. putida cells were immobilized in calcium alginate beads. the degradation of acetonitrile by the immobilized cells of p. putida was investigated. the immobilized cells degraded different concentrations of acetonitrile into ammonia and carbon dioxide. the effect of aeration on the degradation rate was also studi ...	1993	8323268
cloning and nucleotide sequence analysis of the ferripyoverdine receptor gene fpva of pseudomonas aeruginosa.	pseudomonas aeruginosa k437 lacks the ferripyoverdine receptor and, as a result, grows poorly on an iron-deficient minimal medium supplemented with ethylenediamine-di(o-hydroxyphenylacetic acid) (eddha) and pyroverdine. by using a phagemid-based in vivo cloning system, attempts were made to clone the receptor gene by complementing this growth defect. several recombinant phagemids carrying p. aeruginosa chromosomal dna which provided for good growth on eddha-pyoverdine-containing medium and which ...	1993	8335619
in vivo reactivation of catechol 2,3-dioxygenase mediated by a chloroplast-type ferredoxin: a bacterial strategy to expand the substrate specificity of aromatic degradative pathways.	the meta-cleavage operon of the tol plasmid pww0 of pseudomonas putida contains 13 genes responsible for the oxidation of benzoate and toluates to krebs cycle intermediates via estradiol (meta) cleavage of (methyl)catechol. the functions of all the genes are known with the exception of xylt. we constructed pww0 mutants defective in the xylt gene, and found that these mutants were not able to grow on p-toluate while they were still capable of growing on benzoate and m-toluate. in the xylt mutants ...	1993	8344270
the pseudomonas putida ml2 plasmid-encoded genes for benzene dioxygenase are unusual in codon usage and low in g+c content.	benzene dioxygenase, catalyzing the oxidation of benzene to cis-1,2-dihydroxy-cyclohexa-3,5-diene, comprises four polypeptides that are encoded by plasmid phmt112 of pseudomonas putida ml2. in this study, the nucleotide (nt) sequences of four genes encoding this enzyme (bedc1c2ba) were determined, and the amino acid (aa) sequences were deduced. the sequence showed significant homology with the chromosomally encoded benzene dioxygenase and toluene dioxygenase genes (73-77% for nt and 83-99% for a ...	1993	8344526
gene components responsible for discrete substrate specificity in the metabolism of biphenyl (bph operon) and toluene (tod operon).	bph operons coding for biphenyl-polychlorinated biphenyl degradation in pseudomonas pseudoalcaligenes kf707 and pseudomonas putida kf715 and tod operons coding for toluene-benzene metabolism in p. putida f1 are very similar in gene organization as well as size and homology of the corresponding enzymes (g. j. zylstra and d. t. gibson, j. biol. chem. 264:14940-14946, 1989; k. taira, j. hirose, s. hayashida, and k. furukawa, j. biol. chem. 267:4844-4853, 1992), despite their discrete substrate rang ...	1993	8349562
isolation and preliminary characterization of the subunits of the terminal component of naphthalene dioxygenase from pseudomonas putida ncib 9816-4.	the terminal oxygenase component (ispnap) of naphthalene dioxygenase from pseudomonas putida ncib 9816-4 was purified to homogeneity. the protein contained approximately 4 g-atoms each of iron and acid-labile sulfide per mol of ispnap, and enzyme activity was stimulated significantly by addition of exogenous iron. the large (alpha) and small (beta) subunits of ispnap were isolated by two different procedures. the nh2-terminal amino acid sequences of the alpha and beta subunits were identical to ...	1993	8376335
reinvestigation of the role of thiol groups of glyoxalase i purified from yeast (saccharomyces cerevisiae).	glyoxalase i has been purified to homogeneity from saccharomyces cerevisiae and tested with two different thiol reagents, i.e., 5,5'-dithiobis-(2-nitrobenzoic acid) (dtnb) and 1-chloro-2,4-dinitrobenzene (cdnb). dtnb reacts with four thiol groups per molecule of enzyme and leads to a complete inhibition which is not reversed by addition of the disulfide-reducing agent dithiothreitol. on the other hand, cdnb slightly affects the glyoxalase-i activity and alkylates only one thiol residue/enzyme. i ...	1993	8373819
engineering of alkyl- and haloaromatic-responsive gene expression with mini-transposons containing regulated promoters of biodegradative pathways of pseudomonas.	four recombinant mini-tn5 transposons are described which contain outward-facing pm, pu or psal promoters from the catabolic plasmids tol and nah of pseudomonas putida, along with their cognate wild-type regulatory genes (xyls, xylr, nahr) or mutant varieties (xyls2). transcription from such promoters is activated when the host bacteria encounters certain aromatic compounds, such as alkyl- and halobenzoates (xyls, xyls2), alkyl- and halotoluenes (xylr) or salicylates (nahr). these transposons en ...	1993	8393826
regulation of the pcaij genes for aromatic acid degradation in pseudomonas putida.	six of the genes encoding enzymes of the beta-ketoadipate pathway for benzoate and 4-hydroxybenzoate degradation in pseudomonas putida are organized into at least three separate transcriptional units. as an initial step to defining this pca regulon at the molecular level, lacz fusions were made with the pcai and pcaj genes, which encode the two subunits of beta-ketoadipate:succinyl-coenzyme a transferase, the enzyme catalyzing the next-to-last step in the beta-ketoadipate pathway. fusion analyse ...	1993	8376330
heteronuclear nmr analysis of unsaturated fatty acids in poly(3-hydroxyalkanoates). study of beta-oxidation in pseudomonas putida.	poly(3-hydroxyalkanoates) (phas) were isolated from pseudomonas putida kt2442 cultivated on petroselenic acid, oleic acid, and linoleic acid to study beta-oxidation of unsaturated fatty acids. both saturated and unsaturated medium chain length 3-hydroxy fatty acids were found to be constituents of these polymers. with the aid of proton-detected multiple quantum coherence and proton-detected multiple bond coherence nmr spectra the structures of the unsaturated monomers were identified as 3-hydrox ...	1993	8416939
transformation of 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8- dihydroxyquinoline by quinoline-grown resting cells of pseudomonas putida 86.	resting cells of pseudomonas putida strain 86 were grown on quinoline transformed 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8-dihydroxyquinoline which was not converted further. 7,8-dioxygenating activity was present when the enzymes of quinoline catabolism were induced. quinoline-grown cells of strain 86 treated simultaneously with 2-chloroquinoline and d-(-)-threo-chloramphenicol to prevent protein biosynthesis also formed the cis-7,8-dihydrodiol of 2-chloroquinoline. succinate-grown res ...	1993	8405957
cbbr, a lysr-type transcriptional activator, is required for expression of the autotrophic co2 fixation enzymes of xanthobacter flavus.	xanthobacter flavus is able to grow autotrophically with the enzymes of the calvin cycle for the fixation of co2, which are specified by the cbblsxfp gene cluster. previously, the 5' end of an open reading frame (cbbr), displaying a high sequence similarity to the lysr family of regulatory proteins and transcribed divergently from cbblsxfp, was identified (w. g. meijer, a. c. arnberg, h. g. enequist, p. terpstra, m. e. lidstrom, and l. dijkhuizen, mol. gen. genet. 225:320-330, 1991). this paper ...	1993	8407781
cloning, sequencing, and expression of the pseudomonas putida protocatechuate 3,4-dioxygenase genes.	the genes that encode the alpha and beta subunits of protocatechuate 3,4-dioxygenase (3,4-pcd [ec 1.13.11.3]) were cloned from a pseudomonas putida (formerly p. aeruginosa) (atcc 23975) genomic library prepared in lambda phage. plaques were screened by hybridization with degenerate oligonucleotides designed using known amino acid sequences. a 1.5-kb smai fragment from a 15-kb primary clone was subcloned, sequenced, and shown to contain two successive open reading frames, designated pcah and pcag ...	1993	8407791
nucleotide sequence and initial functional characterization of the clcr gene encoding a lysr family activator of the clcabd chlorocatechol operon in pseudomonas putida.	the 3-chlorocatechol operon clcabd is central to the biodegradative pathway of 3-chlorobenzoate. the clcr regulatory gene, which activates the clcabd operon, was cloned from the region immediately upstream of the operon and was shown to complement an insertion mutation for growth on 3-chlorobenzoate. clcr activated the clca promoter, which controls expression of the clcabd operon, in trans by 14-fold in an in vivo promoter probe assay in pseudomonas putida when cells were incubated with 15 mm 3- ...	1993	8419291
proteins induced by sulfate limitation in escherichia coli, pseudomonas putida, or staphylococcus aureus.	two-dimensional gel electrophoresis of proteins from escherichia coli, pseudomonas putida, and staphylococcus aureus, grown with methionine or one of a variety of organosulfates and organosulfonates as the sole source of sulfur, showed expression of specific sets of 7 to 14 proteins which were not observed during growth with sulfate or cysteine for all three species or with thiocyanate for p. putida and s. aureus. under the same conditions, arylsulfatase activity in p. putida and s. aureus was s ...	1993	8432711
identification and characterization of the exbb, exbd and tonb genes of pseudomonas putida wcs358: their involvement in ferric-pseudobactin transport.	catechol-cephalosporins are siderophore-like antibiotics which are taken up by cells of pseudomonas putida wcs358 via the ferric-siderophore transport pathway. mutants of strain wcs358 were isolated that are resistant to high concentrations of these antibiotics. these mutants failed to grow under iron-limiting conditions, and could not utilize different ferric-siderophores. the mutants fall in three complementation groups. the nucleotide sequence determination identified three contiguous open re ...	1993	8437515
precise deletions in large bacterial genomes by vector-mediated excision (vex). the trfa gene of promiscuous plasmid rk2 is essential for replication in several gram-negative hosts.	we have developed a simple and efficient method of vector-mediated excision (vex) for in vivo generation of precisely defined deletions in large bacterial genomes. the system uses homologous recombination with small cloned fragments on specialized pvex plasmids to insert directly repeated bacteriophage p1 loxp sites at positions flanking the region to be deleted. in the presence of cre recombinase, the loxp sites are efficiently recombined to produce the deletion. deletion endpoints can be direc ...	1993	8450534
construction of a pseudomonas hybrid strain that mineralizes 2,4,6-trinitrotoluene.	a bacterium, pseudomonas sp. strain c1s1, able to grow on 2,4,6-trinitrotoluene (tnt), 2,4- and 2,6-dinitrotoluene, and 2-nitrotoluene as n sources, was isolated. the bacterium grew at 30 degrees c with fructose as a c source and accumulated nitrite. through batch culture enrichment, we isolated a derivative strain, called pseudomonas sp. clone a, which grew faster on tnt and did not accumulate nitrite in the culture medium. use of tnt by these two strains as an n source involved the successive ...	1993	8468288
cloning, sequencing, and genetic characterization of regulatory genes, rina and rinb, required for the activation of staphylococcal phage phi 11 int expression.	the int gene of staphylococcal bacteriophage phi 11 is the only viral gene responsible for the integrative recombination of phi 11. to study the regulation of int gene expression, we determined the 5' end of the transcript by s1 mapping. the presumed promoter is located just 22 nucleotides upstream of the int open reading frame in a region which is conserved between phi 11 and a closely related staphylococcal phage, l54a. to clone the possible regulatory gene, a vector which contained the report ...	1993	8432703
enhanced mineralization of polychlorinated biphenyls in soil inoculated with chlorobenzoate-degrading bacteria.	an altamont soil containing no measurable population of chlorobenzoate utilizers was examined for the potential to enhance polychlorinated biphenyl (pcb) mineralization by inoculation with chlorobenzoate utilizers, a biphenyl utilizer, combinations of the two physiological types, and chlorobiphenyl-mineralizing transconjugants. biphenyl was added to all soils, and biodegradation of 14c-aroclor 1242 was assessed by disappearance of that substance and by production of 14co2. mineralization of pcbs ...	1993	8476293
sequences of genes encoding naphthalene dioxygenase in pseudomonas putida strains g7 and ncib 9816-4.	the multicomponent enzyme, naphthalene dioxygenase, initiates the metabolism of naphthalene by pseudomonas putida strains g7 (ppg7) and ncib 9816-4 (pp9816-4). the genes involved (nahaaabacad) are encoded by the nah7 and pdtg1 plasmids, respectively, and form part of the nah operon. the locations of the structural genes were determined on previously cloned fragments of dna. the nucleotide (nt) sequences were determined for nahaaab from pp9816-4 and for nahaaabacad from ppg7. the appropriate open ...	1993	8486285
the amino acid sequence of pseudomonas putida azurin.	the low molecular weight "blue" copper protein, azurin, has been purified from pseudomonas putida (ncib 9869) to homogeneity using various chromatographic techniques including reverse-phase hplc. the amino acid sequence of the n-terminus of the reduced and carboxymethylated protein yielded a single sequence corresponding to aeckv. the complete sequence, comprising 128 amino acid residues with a c-terminal sequence corresponding to tvtlk, was determined from the peptides obtained from a staphyloc ...	1993	8489263
oxidation of aniline to nitrobenzene by nonheme bromoperoxidase.	nonheme bromoperoxidase found in pseudomonas putida catalyzed the bromination of aniline with hydrogen peroxide and bromide ions to give o- and p-bromoanilines. however, in the absence of bromide ions, it oxidized aniline via azobenzene and azoxybenzene finally into nitrobenzene. this is the first report of the biological oxidation of an arylamine to the corresponding nitrocompound at the enzyme level. in addition, nitrobenzene was not formed by a nonheme bromoperoxidase from corallina pilulifer ...	1993	8490583
kinetic studies on benzyl alcohol dehydrogenase encoded by tol plasmid pwwo. a member of the zinc-containing long chain alcohol dehydrogenase family.	the nucleotide sequence of the structural gene for benzyl alcohol dehydrogenase encoded by tol plasmid pwwo of pseudomonas putida has been determined. benzyl alcohol dehydrogenase is a member of the long-chain zinc alcohol dehydrogenase family and, like other alcohol dehydrogenases of this family, contains two zinc atoms per subunit. benzyl alcohol dehydrogenase, while sharing 31% identical residues with horse liver alcohol dehydrogenase, contains several amino acid substitutions near the active ...	1993	8496150
molecular analysis of the plasmid-borne bed gene cluster from pseudomonas putida ml2 and cloning of the cis-benzene dihydrodiol dehydrogenase gene.	pseudomonas putida ml2 contains a large catabolic plasmid, phmt112, carrying genes that encode the dioxygenase and dehydrogenase involved in the catabolism of benzene via the ortho or beta-ketoadipate pathway. phmt112 was derived from a larger and less stable plasmid in p. putida ml2 following growth on succinate as carbon and energy source but was, however, stably maintained in p. putida even in the absence of selection for growth on benzene. cleavage sites for the restriction endonucleases dra ...	1993	8500007
superoxide dismutase activity in root-colonizing pseudomonads.	several saprophytic fluorescent pseudomonads that are aggressive root colonizers express similar specific activities of superoxide dismutase during growth in liquid culture. the pseudomonads have the potential to produce hydrogen peroxide sensitive and hydrogen peroxide insensitive isoforms of superoxide dismutase with distinct mobilities in nondenaturing polyacrylamide gel electrophoresis. synthesis of the hydrogen peroxide insensitive form is enhanced by limited iron availability, by exposure ...	1993	8500011
creatinase in its collapsed a state shows properties of a molten globule with dimeric quaternary structure.	in the past, the molten globule state at acidic ph (a state) has mainly been observed for small single-domain proteins. for more complex proteins such as immunoglobulin, alternatively folded states, with certain characteristics of the molten globule but different thermodynamic properties, were observed. in the present work, the acid-induced structural characteristics of a homodimer, creatinase from pseudomonas putida, are described. the 91-kda protein at ph 2 shows molten globule behavior in tha ...	1993	8504814
physical organization of the upper pathway operon promoter of the pseudomonas tol plasmid. sequence and positional requirements for xylr-dependent activation of transcription.	the upper pathway operon of the pseudomonas putida tol plasmid belongs to the -12/-24 class of promoters. these promoters exhibit three regions critical for regulated transcription, namely, the -12/-24 site for rna polymerase/sigma 54 binding, the -55/-67 region for ihf protein binding, and the -130(uas2)/-170(uas1) region, where two sites for xylr binding are located. the xylr-protected g residues located at -131, -139, -160 and -169 were replaced with as, and the activity of the mutant promote ...	1993	8510657
comparison of the nucleotide sequences of the meta-cleavage pathway genes of tol plasmid pww0 from pseudomonas putida with other meta-cleavage genes suggests that both single and multiple nucleotide substitutions contribute to enzyme evolution.	tol plasmid pww0 from pseudomonas putida mt-2 encodes catabolic enzymes required for the oxidation of toluene and xylenes. the structural genes for these catabolic enzymes are clustered into two operons, the xylcmabn operon, which encodes a set of enzymes required for the transformation of toluene/xylenes to benzoate/toluates, and the xylxyzltegfjqkih operon, which encodes a set of enzymes required for the transformation of benzoate/toluates to krebs cycle intermediates. the latter operon can be ...	1993	8510667
diketocamphane enantiomer-specific 'baeyer-villiger' monooxygenases from camphor-grown pseudomonas putida atcc 17453.	pseudomonas putida atcc 17453 grew with either (+)- or (-)-camphor as sole carbon source. enantiomer-specific 'biological baeyer-villiger' monooxygenases were synthesized irrespective of the camphor isomer used for growth. the two enzymes are probably the products of separate genes but showed many similarities. each consisted of two electrophoretically identical subunits, bound flavin mononucleotide (fmn) non-covalently and accepted electrons from an induced nadh dehydrogenase which interacted w ...	1993	8515237
development of bacterial cytochrome p-450(cam) (cytochrome m) production.	cytochrome p-450(cam) monooxygenase is an important bacterial redox enzyme system with potential commercial value for detoxifying trace hydrocarbon contaminants, catalyzing regiospecific hydroxylations, and amperometric biosensing. the present study was undertaken to increase productivity of this enzyme, which is induced in its host, pseudomonas putida ppg 786, by d(+)-camphor. culture processes were studied in batch, fed-batch, and continuous modes to evaluate metabolic behavior and develop con ...	1993	18609569
phenol removal from waste gases with a biological filter by pseudomonas putida.	the purpose of this study is to investigate the feasibility of biologically removing phenol from waste gases by means of a biofilter using a pseudomonas putida strain. two series of both batch and continuous tests have been performed in order to ascertain the microbial degradation of phenol. for the preliminary batch tests, carried out in order to test the effective feasibility of the process and to investigate their kinetic behavior, two different microbial cultures belonging to the pseudomonas ...	1993	18609611
broad host range, regulated expression system utilizing bacteriophage t7 rna polymerase and promoter.	an iptf-regulated broad host range expression system was constructed using compatible broad host range plasmids, the t7 rna polymerase, and t7 promoter sequences. the system is implemented by the coexistence of two plasmids. the first contains the t7 rna polymerase gene under the control of lacl or lacl(q) genes and lacuv5 promoter. the second encodes the t7 promoter upstream of a multicloning site. incp1 or incp4 t7 promoter plasmids, and incp1, incp4 or incw t7 rna polymerase plasmids were con ...	1993	18609631
competition between two microbial populations in a sequencing fed-batch reactor: theory, experimental verification, and implications for waste treatment applications.	competition between two microbial populations for a single pollutant (phenol) was studied in a sequencing fed-batch reactor (sfbr). a mathematical model describing this system was developed and tested experimentally. it is based on specific growth rate expressions revealed from pure culture batch experiments. the species employed were pseudomonas putida (atcc 17514) and pseudomonas resinovorans (atcc 14235). it was found that both species biodegrade phenol following inhibitory kinetics which can ...	1993	18613087
dynamics of phenol degradation by pseudomonas putida.	pure cultures of pseudomonas putida (atcc 17484) were grown in continuous culture on phenol at dilution rates of 0.074-0.085 h(-1) and subjected to step increases in phenol feed concentration. three distinct patterns of dynamic response were obtained depending on the size of the step change used: low level, moderate level, or high level. during low level responses no accumulations of phenol or non-phenol, non-glucose-dissolved organic carbon, doc(ngp), were observed. moderate level responses wer ...	1993	18609589
biological removal of inorganic hg(ii) as gaseous elemental hg(0) by continuous culture of a hg-resistant pseudomonas putida strain fb-1.	a strain of broad-spectrum, mercury-resistant pseudomonas putida fb1 was used to remove mercury as the gaseous element (hg(0)) from a continuous axenic culture, fed with a synthetic medium containing 1 mg hg l(-1) as hgcl2. mercury determinations were performed in steady-state cultures using various culture fractions [whole culture, filtered supernatant, bacterial cells (dry wt), recovery trap liquid] in order to determine the removal efficiency at different dilution rates (from 0.1 to 3.0 day(- ...	1993	24419964
degradation of nitriles and amides by the immobilized cells of pseudomonas putida.	pseudomonas putida, capable of utilizing acetonitrile as a sole source of c and n, was immobilized in calcium alginate and the rates of degradation of nitriles, including acetonitrile, and their respective amides were studied. all the organic nitriles and amides tested were converted into nh3 and co2.	1993	24420117
gene expression in pseudomonas.	gene regulation studies in pseudomonad bacteria are mainly restricted to pseudomonas aeruginosa and pseudomonas putida. constitutive promoters exhibit dna sequences similar to the σ (70)-dependent constitutive promoters of escherichia coli. the tol meta-cleavage pathway operon promoter and the nah operon promoters are the best characterized σ (70)-dependent promoters, which exhibit-10 regions rich in as and ts and non-conserved-35 regions. the dna binding motif recognized by the respective posit ...	1993	24420110
degradation of delor 103, a technical mixture of polychlorinated biphenyls, by selected bacteria.	ability to utilize a technical mixture of polychlorinated biphenyls (pcb), delor 103, as the sole carbon source, has been tested in 14 bacterial strains. for the five best growing strains (alcaligenes latus, alcalgenes eutrophus, comamonas testosteroni, micrococcus varians and pseudomonas putida), the dependence of the degradation of individual pcb congeners on the number of chlorine substituents is discussed.	1993	24420291
hydrolysis of triglyceride by the whole cell of pseudomonas putida 3sk in two-phase batch and continuous reactors systems.	batch and continuous hydrolysis of olive oil in an organic-aqueous two-phase system using the live whole cell of pseudomonas putida 3sk as a source of a lipase is investigated. the strain was not only fully viable and grown well, but also produced extracellular lipase simultaneously. the degree of hydrolysis, depending on olive oil concentration in the solvents, was maximal at 13.5% (w/v) and decreased with the increase of the substrate concentration. at the optimal condition, a degree of hydrol ...	1994	18618777
biodegradation process development using a bacterial cytochrome in vivo.	pseudomonas putida ppg786 that contains the inducible enzyme system cytochrome p-450(cam) is considered for use as specialized biomass fore detoxification of hazardous hydrocarbons. the test substrate 1,2-dibromochloropropane (dbcp) is used to assess the organohalide degradation activity of p. putida ppg786. activity was found to be a strong function of intracellular heme content, variables which affect the culturing and processing of the cells, and oxygen tension in the degradation incubation m ...	1994	18618691
metabolic engineering of pseudomonas putida for the simultaneous biodegradation of benzene, toluene, and p-xylene mixture.	for the complete biodegradation of a mixture of benzene, toluene, and p-xylene (btx), a critical metabolic step that can connect two existing metabolic pathways of aromatic compounds (the tod and the tol pathways) was determined. toluate-cis-glycol dehydrogenase in the tol pathway was found to attack benzene-cis-glycol, toluene-cis-glycol, and p-xylene-cis-glycol, which are metabolic intermediates of the tod pathway. based on this observation, a hybrid strain, pseudomonase putida tb101, was cons ...	1994	18615528
passive protection of diabetic rats with antisera specific for the polysaccharide portion of the lipopolysaccharide isolated from pseudomonas pseudomallei.	polyclonal and monoclonal antisera raised to tetanus toxoid-conjugated polysaccharide of lipopolysaccharide (lps) and purified lps of pseudomonas pseudomallei that reacted with a collection of 41 strains of this bacterium from 23 patients are described. the common antigen recognized by these sera was within the polysaccharide component of the lps of the cells. the sera were specific for p pseudomallei in that none of 37 strains of other bacteria, including 20 gram-negative and three gram-positiv ...	1994	22346496
the role of lysine 166 in the mechanism of mandelate racemase from pseudomonas putida: mechanistic and crystallographic evidence for stereospecific alkylation by (r)-alpha-phenylglycidate.	the mechanism of irreversible inactivation of mandelate racemase (mr) from pseudomonas putida by alpha-phenylglycidate (alpha pga) has been investigated stereochemically and crystallographically. the (r) and (s) enantiomers of alpha pga were synthesized in high enantiomeric excess (81% ee and 83% ee, respectively) using sharpless epoxidation chemistry. (r)-alpha pga was determined to be a stereospecific and stoichiometric irreversible inactivator of mr. (s)-alpha pga does not inactivate mr and a ...	1994	8292591
fpta, the fe(iii)-pyochelin receptor of pseudomonas aeruginosa: a phenolate siderophore receptor homologous to hydroxamate siderophore receptors.	the pseudomonas aeruginosa siderophore pyochelin is structurally unique among siderophores and possesses neither hydroxamate- nor catecholate-chelating groups. the structural gene encoding the 75-kda outer membrane fe(iii)-pyochelin receptor fpta has been isolated by plasmid rescue techniques and sequenced. the n-terminal amino acid sequence of the isolated fpta protein corresponded to that deduced from the nucleotide sequence of the fpta structural gene. the mature fpta protein has 682 amino ac ...	1994	8288523
putative functions of phenylalanine-350 of pseudomonas putida cytochrome p-450cam.	cytochrome p-450cam hydroxylates d-camphor, using molecular oxygen and reducing equivalents transferred via putidaredoxin. we constructed mutant genes in which phe-350 of p-450cam was replaced by leu, tyr, or his by site-directed mutagenesis, expressed them in escherichia coli, purified the mutant proteins, and compared their enzymic properties with those of the wild type p-450cam. nadh oxidation rate of the tyr mutant in the reconstituted system with putidaredoxin and putidaredoxin reductase wa ...	1994	8305479
genetic evidence that the xyls regulator of the pseudomonas tol meta operon controls the pm promoter through weak dna-protein interactions.	the activation of the pm promoter of the meta operon of the tol plasmid of pseudomonas putida by its cognate xyls activator protein in the presence and absence of benzoate inducers has been examined in specialized escherichia coli strains carrying pm-lacz fusions and the xyls gene in different configurations in which all controlling elements are present in near native conditions and stoichometry. expression of a chromosomal pm-xylx::lacz fusion was primarily dependent on the addition of an effec ...	1994	8195070
oxidation of low molecular weight chloroalkanes by cytochrome p450cam.	cytochrome p450cam from pseudomonas putida g786 oxidized chlorinated ethanes and 1,2-dichloropropane. the rate of nadh oxidation decreased with decreasing chlorine substitution. the single detectable oxidation products of 1,1,1-trichloroethane and 1,2-dichloropropane were 1,1,1-trichloroethanol and chloroacetone, respectively. organic product formation was largely uncoupled from nadh oxidation.	1994	8198597
molecular characterization of microbial alcohol dehydrogenases.	there is an astonishing array of microbial alcohol oxidoreductases. they display a wide variety of substrate specificities and they fulfill several vital but quite different physiological functions. some of these enzymes are involved in the production of alcoholic beverages and of industrial solvents, others are important in the production of vinegar, and still others participate in the degradation of naturally occurring and xenobiotic aromatic compounds as well as in the growth of bacteria and ...	1994	8185833
secondary 18o and primary 13c isotope effects as a probe of transition-state structure for enzymatic decarboxylation of oxalacetate.	a new method for directly measuring 18o isotope effects on decarboxylation reactions has been developed. by running the reaction under high vacuum (10(-5) torr), co2 leaves the solution before exchange with the oxygens of water to an extent greater than 2%. thus, the method permits determination of 18o isotope effects with the precision of the isotope ratio mass spectrometer, and without the necessity of resorting to the remote label method and its attendant required syntheses. the method is use ...	1994	8172901
mechanism of p-hydroxyphenylacetate-3-hydroxylase. a two-protein enzyme.	p-hydroxyphenylacetate-3-hydroxylase purified from pseudomonas putida is a two-protein enzyme requiring a flavoprotein and a coupling protein for productive hydroxylation (arunachalam, u., massey, v., and vaidyanathan, c. s. (1992) j. biol. chem. 267, 25848-25855). this paper presents information on the mechanism of the enzyme from absorbance and fluorescence stopped-flow studies. the reduction of the substrate-free flavoprotein by nadh was slow and was not altered by the presence of the couplin ...	1994	8276789
responses to nutrient starvation in pseudomonas putida kt2442: analysis of general cross-protection, cell shape, and macromolecular content.	the physiology of pseudomonas putida kt2442 with respect to growth and carbon starvation was studied. during the transition from growth to nongrowth, the cell shape changes from cylindrical to spheric, a change which is accompanied by reductions in cell size, dna and ribosome content, and the rate of total protein synthesis. in addition, a pattern of general cross-protection develops, which enables the cells to survive environmental stresses such as high and low temperatures, elevated osmolarity ...	1994	8282712
cloning and high-level expression of the glutathione-independent formaldehyde dehydrogenase gene from pseudomonas putida.	a dna fragment of 485 bp was specifically amplified by pcr with primers based on the n-terminal sequence of the purified formaldehyde dehydrogenase (ec 1.2.1.46) from pseudomonas putida and on that of a cyanogen bromide-derived peptide. with this product as a probe, a gene coding for formaldehyde dehydrogenase (fdha) in p. putida chromosomal dna was cloned in escherichia coli dh5 alpha. sequencing analysis revealed that the fdha gene contained 1,197-bp open reading frame, encoding a protein comp ...	1994	8169197
transcriptional induction kinetics from the promoters of the catabolic pathways of tol plasmid pww0 of pseudomonas putida for metabolism of aromatics.	we determined, under several growth conditions, the kinetics of mrna synthesis from the four pseudomonas putida pww0 plasmid promoters involved in degradation of xylenes and methylbenzyl alcohols via toluates. transcription by xyls of the meta-cleavage pathway operon promoter (pm) for the metabolism of alkylbenzoates was stimulated immediately after the addition of an effector, both in luria-bertani (lb) medium and in minimal medium. activation of the sigma 54-dependent upper-pathway operon prom ...	1994	8169200
aliphatic and aromatic inhibitors binding to the active site of catechol 2,3-dioxygenase from pseudomonas putida mt-2.	the interaction of different classes of inhibitors with the extradiol cleaving catechol 2,3-dioxygenase from pseudomonas putida mt-2 was monitored by longitudinal and transverse proton relaxation measurements as well as by kinetic activity studies in order to characterize the type of interaction of such inhibitors with the active site of the enzyme. the average distances of the inhibitors from the catalytic iron(ii) ion have been estimated from the 1h longitudinal relaxation rates. phenols and a ...	1994	8163017
purification of the lysr family regulator, clcr, and its interaction with the pseudomonas putida clcabd chlorocatechol operon promoter.	previous studies have shown that the clcabd operon is under the transcriptional control of the lysr-type activator clcr. in this study, the conditions leading to its aggregation were avoided and clcr was purified and confirmed by amino-terminal sequencing. gel filtration indicated that clcr exists as a dimer in solution. the dnase i footprint of clcr was determined. the binding properties of clcr and the catechol operon regulator, catr, were compared.	1994	8071232
identification and characterization of a transmissible linear plasmid from rhodococcus erythropolis bd2 that encodes isopropylbenzene and trichloroethene catabolism.	rhodococcus erythropolis bd2, which is able to utilize isopropylbenzene as a sole carbon and energy source, was shown to contain a conjugative linear plasmid, pbd2. the estimated size of pbd2 is 208 to 212 kb. linear plasmid-deficient strains had lost both the isopropylbenzene degradation and trichloroethene degradation characteristics, as well as the arsenite resistance and mercury resistance phenotypes. reintroduction of pbd2 restored all four characteristics. conjugational transfer of pbd2 to ...	1994	8161179
catabolism of aromatics in pseudomonas putida u. formal evidence that phenylacetic acid and 4-hydroxyphenylacetic acid are catabolized by two unrelated pathways.	phenylacetic acid (phacoh) and 4-hydroxyphenylacetic acid (4hophacoh) are catabolized in pseudomonas putida u through two different pathways. mutation carried out with the transposon tn5 has allowed the isolation of several mutants which, unlike the parental strain, are unable to grow in chemically defined medium containing either phacoh or 4hophacoh as the sole carbon source. analysis of these strains showed that the ten mutants unable to grow in phacoh medium grew well in the one containing 4h ...	1994	8168524
sequence and structure comparison suggest that methionine aminopeptidase, prolidase, aminopeptidase p, and creatinase share a common fold.	amino acid sequence comparison suggests that the structure of escherichia coli methionine aminopeptidase (ec 3.4.11.18) and the c-terminal domain of pseudomonas putida creatinase (ec 3.5.3.3) are related. a detailed comparison of the three-dimensional folds of the two enzymes confirms this homology: with an approximately 260-residue chain segment, 218 c alpha atoms of the structures superimpose within 2.5 a; only 41 of these overlapping positions (i.e., 19%) feature identical amino acids in the ...	1994	8146141
crystallization of catechol-1,2 dioxygenase from pseudomonas arvilla c-1.	the metalloenzyme catechol 1,2-dioxygenase from pseudomonas arvilla c-1 consists of three isozymes formed by combinations of two non-identical subunits; alpha alpha, alpha beta and beta beta; with molecular masses of 59,000, 63,000 and 67,000 da, respectively. the alpha alpha isozyme crystallizes in the orthorhombic space group c222(1) with unit cell dimensions a = 62.7 a, b = 71.5 a, c = 187.1 a. the rectangular plates diffract to 2.6 a resolution. this is the first dioxygenase to be crystalliz ...	1994	8107120
cloning and characterization of a chromosomal gene cluster, pah, that encodes the upper pathway for phenanthrene and naphthalene utilization by pseudomonas putida ous82.	a 25-kb dna sali fragment cloned from the chromosomal dna of pseudomonas putida ous82, which utilizes phenanthrene (phn+) and naphthalene (nah+), carried all of the genes necessary for upper naphthalene catabolism. cosmid recombinant pip7 complemented both the nah- and phn- defects of ous8211 (trp-nah-phn-sal+[salicylate utilizing]hna+[1-hydroxy-2-naphthoate utilizing]) and only the phn- defect of ous8212 (trp-nah-phn-sal-hna+). the results indicate that strain ous82 uses different pathways afte ...	1994	8157614
identification of serine-143 as the most likely precursor of dehydroalanine in the active site of histidine ammonia-lyase. a study of the overexpressed enzyme by site-directed mutagenesis.	the gene coding for histidase (histidine ammonia-lyase, hal, ec 4.3.1.3) was isolated from a lambda-embl3 genomic library from pseudomonas putida nicii and subcloned into the expression vector pt7-7. transformation of escherichia coli bl21 (de3) cells with the recombinant vector led to the expression of catalytically active histidase amounting to 20-30% of the total soluble protein in the crude cell extract. a new rapid and highly efficient isolation procedure is described leading to electrophor ...	1994	8204579
molecular analysis of the erwinia chrysanthemi region containing the kdga and zwf genes.	the pathways of pectin and galacturonate catabolism in erwinia chrysanthemi converge to form a common intermediate, 2-keto-3-deoxygluconate, which is phosphorylated to form 2-keto-3-deoxy-6-phosphogluconate (kdgp) and then cleaved by the aldolase encoded by the kdga gene. we cloned the kdga gene of the e. chrysanthemi strain 3937 by complementing an escherichia coli kdga mutation, using an rp4-derivative plasmid. restriction mapping of the kdga region and isolation of kdga-lac fusions allowed th ...	1994	8145647
identification and characterization of genes encoding polycyclic aromatic hydrocarbon dioxygenase and polycyclic aromatic hydrocarbon dihydrodiol dehydrogenase in pseudomonas putida ous82.	naphthalene and phenanthrene are transformed by enzymes encoded by the pah gene cluster of pseudomonas putida ous82. the paha and pahb genes, which encode the first and second enzymes, dioxygenase and cis-dihydrodiol dehydrogenase, respectively, were identified and sequenced. the dna sequences showed that paha and pahb were clustered and that paha consisted of four cistrons, pahaa, pahab, pahac, and pahad, which encode ferredoxin reductase, ferredoxin, and two subunits of the iron-sulfur protein ...	1994	8157615
x-ray absorption studies on catechol 2,3-dioxygenase from pseudomonas putida mt2.	x-ray absorption spectroscopy has been utilized to investigate the structure of the active site of iron(ii) catechol 2,3-dioxygenase from pseudomonas putida mt2 both in the native and the 2-chlorophenol inhibited forms. xanes (x-ray absorption near edge structure) and exafs (extended x-ray absorption fine structure) results allow us to discuss the coordination number and geometry of the ferrous ion in the native enzyme. the metal geometry is not significantly affected by the binding of the inhib ...	1994	8075079
synthesis of trans unsaturated fatty acids in pseudomonas putida p8 by direct isomerization of the double bond of lipids.	the phospholipids of pseudomonas putida p8 contain monounsaturated fatty acids in the cis and trans configuration. cells of this phenol-degrading bacterium change the proportions of these isomers in response to the addition or elimination of a membrane active compound such as 4-chlorophenol. this study undoubtedly reveals that the cis unsaturated fatty acids are directly converted into trans isomers without involvement of de novo synthesis of fatty acids. oleic acid, which cannot be synthesized ...	1994	8085914
cloning and nucleotide sequence of the pseudomonas aeruginosa glucose-selective oprb porin gene and distribution of oprb within the family pseudomonadaceae.	oprb is a glucose-selective porin known to be produced by pseudomonas aeruginosa and pseudomonas putida. we have cloned and sequenced the oprb gene of p. aeruginosa and obtained expression of oprb in escherichia coli. the mature protein consists of 423 amino acid residues with a deduced molecular mass of 47597 da. several clusters of amino acid residues, potentially involved in the structure or function of the protein, were identified. an area of regional homology with e. coli lamb was also iden ...	1994	8125108
an evaluation of molecular models of the cytochrome p450 streptomyces griseolus enzymes p450su1 and p450su2.	p450su1 and p450su2 are herbicide-inducible bacterial cytochrome p450 enzymes from streptomyces griseolus. they have two of the highest sequence indentities to camphor hydroxylase (p450cam from pseudomonas putida), the cytochrome p450 with the first known crystal structure. we have built several models of these two proteins to investigate the variability in the structures that can occur from using different modeling protocols. we looked at variability due to alignment methods, backbone loop conf ...	1994	7876903
regulation of the rpon, orf102 and orf154 genes in pseudomonas putida.	the dna sequence downstream of the pseudomonas putida rpon gene and the adjacent orf102 was determined. this region encodes an orf (orf154) whose gene product was found to be homologous to a family of phosphotransferases. insertional mutagenesis and analysis of mrna transcripts showed that the rpon gene is transcribed separately from the two downstream genes. the rpon promoter was localized to an 86 nucleotide-long region upstream of the rpon gene by examination of the expression of a series of ...	1994	8138132
efficient degradation of trichloroethylene by a hybrid aromatic ring dioxygenase.	engineering of hybrid gene clusters between the toluene metabolic tod operon and the biphenyl metabolic bph operon greatly enhanced the rate of biodegradation of trichloroethylene. escherichia coli cells carrying a hybrid gene cluster composed of todc1 (the gene encoding the large subunit of toluene terminal dioxygenase in pseudomonas putida f1), bpha2 (the gene encoding the small subunit of biphenyl terminal dioxygenase in pseudomonas pseudoalcaligenes kf707), bpha3 (the gene encoding ferredoxi ...	1994	8144482
responses to nutrient starvation in pseudomonas putida kt2442: two-dimensional electrophoretic analysis of starvation- and stress-induced proteins.	the responses of pseudomonas putida kt2442 to various forms of nutrient starvation and stress conditions were examined by two-dimensional polyacrylamide electrophoresis. carbon deprivation resulted in a temporal expression of two classes of starvation-induced proteins: one class was transiently expressed during the initial phase of starvation, and the second class was expressed throughout the entire starvation period. proteins of the second class could be further subdivided into proteins induced ...	1994	8050994
fur regulon in gram-negative bacteria. identification and characterization of new iron-regulated escherichia coli genes by a fur titration assay.	a highly sensitive genetic screen for the detection of cloned genes coding for iron-regulated and iron-storage/binding proteins was developed. the fur titration assay (furta) enabled identification of cloned iron-regulated genes from different gram-positive and gram-negative bacteria such as: bacillus subtilis, escherichia coli, pantoea agglomerans, pseudomonas putida, salmonella typhimurium, serratia marcescens and yersinia enterocolitica. an ordered e. coli cosmid library was screened for eith ...	1994	8107138
cis,cis-muconate lactonizing enzyme from trichosporon cutaneum: evidence for a novel class of cycloisomerases in eucaryotes.	the absolute stereochemical courses of cis,cis-muconate lactonizing enzyme (mle;ec 5.5.1.1) from trichosporon cutaneum (tcmle) and chloromuconate cycloisomerase (mle ii; ec 5.5.1.7) from pseudomonas sp b13 have been determined from 1h nmr measurements. both cycloisomerases convert cis,cis-muconate to (4s)-muconolactone by a syn lactonization, the absolute stereochemical outcome of which is identical to that observed with mle from pseudomonas putida. the regiochemical courses of cyclization of 3- ...	1994	8110801
cross-regulation by xylr and dmpr activators of pseudomonas putida suggests that transcriptional control of biodegradative operons evolves independently of catabolic genes.	the pu promoter of the toluene degradation plasmid pww0 of pseudomonas putida drives expression of an operon involved in the sequential oxidation of toluene and m- and p-xylenes to benzoate and toluates, respectively. similarly, the po promoter of plasmid pvi150 controls expression of an operon of pseudomonas sp. strain cf600 which is required for the complete catabolism of phenol and cresols. these promoters, which both belong to the sigma 54-dependent class, are regulated by their cognate acti ...	1994	8051017
combination effect of meropenem with aminoglycosides and teicoplanin on pseudomonas and enterococci.	the in vitro activity of meropenem, a new carbapenem, and the combination effect with netilmicin, tobramycin, gentamicin, and teicoplanin against pseudomonas spp. and enterococci was studied. meropenem showed very good in vitro activity against pseudomonas aeruginosa (mic90 2 mg/l) and good to moderate activity against pseudomonas putida (mic90 4 mg/l) and enterococcus faecalis (mic90 8 mg/l). aminoglycosides were highly active against p. putida (mic90 0.5 mg/l), but showed only moderate activit ...	1994	8002095
oxidative release of nitrite from 2-nitrotoluene by a three-component enzyme system from pseudomonas sp. strain js42.	pseudomonas sp. strain js42 utilizes 2-nitrotoluene (2nt) as the sole source of carbon and energy for growth. intact cells catalyze the oxidation of 2nt to 3-methylcatechol and nitrite in a reaction that requires molecular oxygen. cell extracts oxidized 2nt to 3-methylcatechol and nitrite in the presence of nad(p)h and ferrous iron. ion-exchange chromatography yielded three protein fractions (a, b, and c) which were all required for the oxidation of 2nt to 3-methylcatechol and nitrite. component ...	1994	8002568
aerobic catabolism of phenylacetic acid in pseudomonas putida u: biochemical characterization of a specific phenylacetic acid transport system and formal demonstration that phenylacetyl-coenzyme a is a catabolic intermediate.	the phenylacetic acid transport system (pats) of pseudomonas putida u was studied after this bacterium was cultured in a chemically defined medium containing phenylacetic acid (pa) as the sole carbon source. kinetic measurement was carried out, in vivo, at 30 degrees c in 50 mm phosphate buffer (ph 7.0). under these conditions, the uptake rate was linear for at least 3 min and the value of km was 13 microm. the pats is an active transport system that is strongly inhibited by 2,4-dinitrophenol, 4 ...	1994	8002592
3-carboxy-cis,cis-muconate lactonizing enzyme from neurospora crassa: an alternate cycloisomerase motif.	3-carboxy-cis,cis-muconate lactonizing enzyme (cmle; ec 5.5.1.5) from neurospora crassa catalyzes the reversible gamma-lactonization of 3-carboxy-cis,cis-muconate by a syn-1,2 addition-elimination reaction. the stereochemical and regiochemical course of the reaction is (i) opposite that of cmle from pseudomonas putida (ec 5.5.1.2) and (ii) identical to that of cis,cis-muconate lactonizing enzyme (mle; ec 5.5.1.1) from p. putida. in order to determine the mechanistic and evolutionary relationship ...	1994	8132467
organization and evolution of naphthalene catabolic pathways: sequence of the dna encoding 2-hydroxychromene-2-carboxylate isomerase and trans-o-hydroxybenzylidenepyruvate hydratase-aldolase from the nah7 plasmid.	the sequence of a 2,437-bp dna segment from the naphthalene upper catabolic pathway operon of plasmid nah7 was determined. this segment contains three large open reading frames designated nahq', nahe, and nahd. the first of these is the 3' end of an open reading frame that has no known function, the second (993 bp) encodes trans-o-hydroxybenzylidenepyruvate hydratase-aldolase (deduced molecular weight, 36,640), and the third (609 bp) encodes 2-hydroxychromene-2-carboxylate isomerase (deduced mol ...	1994	8002605
genetic evidence for activation of the positive transcriptional regulator xy1r, a member of the ntrc family of regulators, by effector binding.	the xy1r protein positively controls expression from the pseudomonas putida tol plasmid sigma 54-dependent pu and ps promoters, in response to the presence of aromatic effectors such as m-xylene, m-methylbenzyl alcohol, and p-chlorobenzaldehyde in the culture medium. xy1r also autoregulates its own synthesis. a mutant xy1r regulator called xy1r7 was isolated after nitrosoguanidine mutagenesis of the wild-type gene and phenotypic selection for mutants that had acquired the ability to recognize m- ...	1994	8132529
codon usage patterns suggest independent evolution of two catabolic operons on toluene-degradative plasmid tol pww0 of pseudomonas putida.	tol plasmid pww0 of pseudomonas putida encodes a set of enzymes responsible for the degradation of toluene. the structural genes for these catobolic enzymes are clustered into two operons--namely, the xy/cmab and xy/xyzltegfjqkih operons. we examined the codon usage patterns of these catabolic genes by measuring the codon-usage distances between pairs of these catabolic genes. the codon-usage distance, d, between gene 1 and gene 2 was defined as d = [sigma(pj-qj)2]1/2, are the frequencies of the ...	1994	8007001