Publications

trichloroethylene removal and oxidation toxicity mediated by toluene dioxygenase of pseudomonas putida.	whole cells of pseudomonas putida containing toluene dioxygenase were able to remove all detectable trichloroethylene (tce) from assay mixtures. the capacity of cells to remove tce was 77 microm/mg of protein with an initial rate of removal of 5.2 nmol/min/ng of protein. tce oxidation resulted in a decrease in the growth rate of cultures and caused rapid cell death. addition of dithiothreitol to assay mixtures increased the tce removal capacity of cells by up to 67% but did not prevent tce-media ...	1994	7811103
chlorocatechol 1,2-dioxygenase from rhodococcus erythropolis 1cp. kinetic and immunochemical comparison with analogous enzymes from gram-negative strains.	chlorocatechol 1,2-dioxygenase from rhodococcus erythropolis 1cp was purified to homogeneity. in contrast to chlorocatechol 1,2-dioxygenase from gram-negative strains which have a very broad substrate tolerance, the rhodococcus enzyme was relatively more specific and had a distinct preference for 4-substituted catechols. protein and metal analysis indicate an unusual stoichiometry of one atom each of iron and manganese/64-kda homodimer. the n-terminal amino acid sequence (27 residues) of the rho ...	1994	7813460
molecular characterization of the pseudomonas putida 2,3-butanediol catabolic pathway.	the 2,3-butanediol dehydrogenase and the acetoin-cleaving system were simultaneously induced in pseudomonas putida ppg2 during growth on 2,3-butanediol and on acetoin. hybridization with a dna probe covering the genes for the e1 subunits of the alcaligenes eutrophus acetoin cleaving system and nucleotide sequence analysis identified acoa (975 bp), acob (1020 bp), apoc (1110 bp), acox (1053 bp) and adh (1086 bp) in a 6.3-kb genomic region. the amino acid sequences deduced from acoa, acob, and aco ...	1994	7813883
[characterization of the fluorescent pigment pyoverdine pm, produced by pseudomonas putida bacteria].	it has been shown that under iron-limited conditions pseudomonas putida m produces large amounts of the fluorescent pigment, pyoverdine pm, which is a siderophore and exhibits antibacterial activity. the absorption spectrum for the pyoverdine pm has two main peaks, at 230 nm and 400 nm, respectively. it was demonstrated that pyoverdine pm molecule besides dihydroxyquinoline moiety has a peptide chain contain five amino acids: threonine, serine, lysine, hydroxyaspartic acid and n delta-hydroxyorn ...	1994	7760765
degradation of methyl parathion by pseudomonas putida.	pseudomonas putida utilized methyl parathion as sole carbon and (or) phosphorus source. the bacterium elaborated the enzyme organophosphorus acid anhydrase, which hydrolyzed methyl parathion to p-nitrophenol. p-nitrophenol was further degraded to hydroquinone and 1,2,4-benzenetriol. the final ring compound, 1,2,4-benzenetriol, was cleaved by benzenetriol oxygenase to maleyl acetate.	1994	7704828
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
[the growth kinetics of pseudomonas putida and its determining factors].		1994	7879508
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
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
regiospecific and stereoselective hydroxylation of 1-indanone and 2-indanone by naphthalene dioxygenase and toluene dioxygenase.	the biotransformation of 1-indanone and 2-indanone to hydroxyindanones was examined with bacterial strains expressing naphthalene dioxygenase (ndo) and toluene dioxygenase (tdo) as well as with purified enzyme components. pseudomonas sp. strain 9816/11 cells, expressing ndo, oxidized 1-indanone to a mixture of 3-hydroxy-1-indanone (91%) and 2-hydroxy-1-indanone (9%). the (r)-3-hydroxy-1-indanone was formed in 62% enantiomeric excess (ee) (r:s, 81:19), while the 2-hydroxy-1-indanone was racemic. ...	1994	7944365
secretion of human epidermal growth factor (egf) in autotrophic culture by a recombinant hydrogen-utilizing bacterium, pseudomonas pseudoflava, carrying broad-host-range egf secretion vector pksegf2.	we constructed the broad-host-range human epidermal growth factor (egf) secretion plasmid pksegf2 by inserting the escherichia coli tac promoter, the signal sequence of pseudomonas stutzeri amylase, and the synthesized egf gene into the broad-host-range vector pkt230. e. coli jm109 carrying pksegf2 secreted egf into the periplasm and the culture medium under the control of the tac promoter. pseudomonas aeruginosa pao1161 carrying pksegf2 and pseudomonas putida ac10 carrying pksegf2 secreted egf ...	1994	7944366
histidine ammonia-lyase mutant s143c is posttranslationally converted into fully active wild-type enzyme. evidence for serine 143 to be the precursor of active site dehydroalanine.	histidase [histidine ammonia-lyase (hal); ec 4.3.1.3] from pseudomonas putida is a homotetramer and contains one catalytically essential dehydroalanine residue per subunit. since the mutant s143a was catalytically inert, it has been proposed that serine 143 is the precursor of the active site dehydroalanine [langer et al. (1994) biochemistry 33, 6462-6467]. to further define the role of serine 143, we prepared the mutants s143t and s143c by site-directed mutagenesis. the threonine 143 mutant was ...	1994	7947813
the gene encoding glyoxalase i from pseudomonas putida: cloning, overexpression, and sequence comparisons with human glyoxalase i.	the gene encoding glyoxalase i (glxi) from pseudomonas putida has been cloned into the high-expression plasmid pbtaci. in the presence of iptg, jm109 cells transformed with this vector give expression levels of glxi 4000-fold higher than wild-type escherichia coli. contrary to a previous report, the nucleotide sequence of the gene encodes a 173-amino-acid polypeptide. edman analysis indicates that the predicted n-terminal methionine is lost post-translationally to yield a 19407-da protein. mass ...	1994	7959071
identification of the pcarkf gene cluster from pseudomonas putida: involvement in chemotaxis, biodegradation, and transport of 4-hydroxybenzoate.	pseudomonas putida prs2000 is chemotactic to 4-hydroxybenzoate and other aromatic acids. this behavioral response is induced when cells are grown on 4-hydroxybenzoate or benzoate, compounds that are degraded via the beta-ketoadipate pathway. isolation of a transposon mutant defective in 4-hydroxybenzoate chemotaxis allowed identification of a new gene cluster designated pcarkf. dna sequencing, mutational analysis, and complementation studies revealed that pcar encodes a regulatory protein requir ...	1994	7961399
conversion of pbr322-based plasmids into broad-host-range vectors by using the tn3 transposition mechanism.	we constructed a series of transposon vectors which allow efficient in vitro gene manipulation and subsequent introduction of cloned dna into a variety of gram-negative bacteria. transfer of the cloned fragment from these multicopy plasmids into self-transmissible broad-host-range vectors is achieved in vivo, using the tn3 transposition mechanism. transposition into a variety of broad-host-range plasmids proceeds efficiently, and the resulting recombinant plasmids can be readily transferred and ...	1994	7961407
cloning, nucleotide sequence, and overexpression of the gene coding for delta 5-3-ketosteroid isomerase from pseudomonas putida biotype b.	the structural gene coding for the delta 5-3-ketosteroid isomerase (ksi) of pseudomonas putida biotype b has been cloned, and its entire nucleotide sequence has been determined by a dideoxynucleotide chain termination method. a 2.1-kb dna fragment containing the ksi gene was cloned from a p. putida biotype b genomic library in lambda gt11. the open reading frame of ksi encodes 393 nucleotides, and the amino acid sequence deduced from the nucleotide sequence agrees with the directly determined am ...	1994	7961420
site-directed mutagenesis of conserved serines in rat histidase. identification of serine 254 as an essential active site residue.	we have identified serine 254 as an essential residue in rat histidase (histidine ammonia-lyase, ec 4.3.1.3). histidase and phenylalanine ammonia-lyase are the only two enzymes that have been postulated to require the modified amino acid, dehydroalanine, for enzyme activity. in the bacterial peptides nisin and subtilin, and in the pyruvoyl enzymes, the precursor for dehydroalanine is a serine. to determine whether serine may be the dehydroalanine precursor in rat histidase, we substituted four h ...	1994	7961661
characterization of type iv pilus genes in plant growth-promoting pseudomonas putida wcs358.	in a search for factors that could contribute to the ability of the plant growth-stimulating pseudomonas putida wcs358 to colonize plant roots, the organism was analyzed for the presence of genes required for pilus biosynthesis. the pild gene of pseudomonas aeruginosa, which has also been designated xcpa, is involved in protein secretion and in the biogenesis of type iv pili. it encodes a peptidase that processes the precursors of the pilin subunits and of several components of the secretion app ...	1994	7905475
cis/trans isomerization of fatty acids as a defence mechanism of pseudomonas putida strains to toxic concentrations of toluene.	defence mechanisms of three pseudomonas putida strains growing in the presence of toluene up to 50% (v/v) were investigated. the three strains reacted to toxic concentrations of toluene by accumulating trans unsaturated fatty acids in the membrane instead of the cis isomers. the membranes of the toluene-adapted cells possessed a higher trans/cis ratio and had a higher lipid-ordering since the transition temperature was about 7 centigrade degrees higher compared to the non-adapted cells.	1994	7921251
molecular cloning of two pseudomonas flagellin genes and basal body structural genes.	pseudomonas putida strains paw8 and prs2000 produce flagellins with apparent molecular masses of 81 kda and 50 kda respectively. two tn5 insertion mutants of p. putida paw8 lacking the ability to bind the flagellin-specific monoclonal antibody mlv1 were isolated. mutant paw8-flg2 contained a tn5 insertion within a 2.6 kb ecori fragment of the p. putida chromosome carrying putative basal body genes. dna and deduced protein sequences suggested the presence on this fragment of two complete genes ho ...	1994	7921252
ecotox-evaluation strategy for soil bioremediation exemplified for a pah-contaminated site.	during a bioremediation of a pah-contaminated site chemical and biological analyses were carried out. the biological investigations included ecotoxicological analyses in the aqueous extract, (pseudomonas putida, photobacterium phosphoreum, daphnids, algae, fish) and analyses in the soil with introduced organisms (plants, earthworms) and natural soil organisms (nematodes, microorganisms). in all test systems a correspondence between decreasing toxicity and degradation of the easily biodegradable ...	1994	7922149
substrate specificity of catechol 2,3-dioxygenase encoded by tol plasmid pww0 of pseudomonas putida and its relationship to cell growth.	catechol 2,3-dioxygenase encoded by tol plasmid pww0 of pseudomonas putida consists of four identical subunits, each containing one ferrous ion. the enzyme catalyzes ring cleavage of catechol, 3-methylcatechol, and 4-methylcatechol but shows only weak activity toward 4-ethylcatechol. two mutants of catechol 2,3-dioxygenases (4ecr1 and 4ecr6) able to oxidize 4-ethylcatechol, one mutant (3mcs) which exhibits only weak activity toward 3-methylcatechol but retained the ability to cleave catechol and ...	1994	7928969
cloning, sequencing, and expression of the structural genes for the cytochrome and flavoprotein subunits of p-cresol methylhydroxylase from two strains of pseudomonas putida.	the structural genes for the flavoprotein subunit and cytochrome c subunit of p-cresol (4-methylphenol) methylhydroxylase (pcmh) from pseudomonas putida ncimb 9869 (national collection of industrial and marine bacteria, aberdeen, scotland) and p. putida ncimb 9866 were cloned and sequenced. the genes from p.putida ncimb 9869 were for the plasmid-encoded a form of pcmh, and the genes from p.putida ncimb 9866 were also plasmid encoded. the nucleotide sequences of the two flavoprotein genes from p. ...	1994	7929007
a revised map location for the histidine utilization genes in pseudomonas putida.	the histidine utilization genes huth and hutu of pseudomonas putida atcc 12633 have been mapped by interrupted mating and transduction to a location at approximately 43 minutes on the chromosome, closely linked to ser-800 and met-400 markers previously shown to be at 46 and 42 minutes, respectively. since restriction enzyme mapping and cloning results have established that all genes associated with the hut pathway are contiguous, earlier maps of this strain which place these genes near 10 minute ...	1994	7932109
heterologous expression of the cytochrome p450cam hydroxylase operon and the repressor gene of pseudomonas putida in escherichia coli.	the cytochrome p450cam hydroxylase operon (camdcab) of pseudomonas putida is negatively regulated by a repressor, camr, which also represses its own gene. the expression in p. putida of both camr and camdcab is derepressed in the presence of d-camphor. we examined the expression in escherichia coli of camr and camdcab by monitoring the enzyme activity of the camd gene product. in the presence or absence of d-camphor in the cell culture, the expression in e. coli of camd was significant and const ...	1994	7988898
[design of the human gm-csf gene using the polymerase chain reaction and its expression in pseudomonas putida cells].	construction of human gm-csf gene was conducted by the pcr technique. four exons of gm-csf gene were synthesized on the basis of human blood dna using thermostable tth dna polymerase. synthetic oligonucleotides were used as primers. the oligonucleotides contained sequences complementary to the ends of exons. joining of exons was conducted by reciprocal complementation of the terminal sequences, followed by filling and amplification of the joined products. in most cases the effective synthesis of ...	1994	7990813
construction of phoe-caa, a novel pcr- and immunologically detectable marker gene for pseudomonas putida.	in this paper we describe the construction and use in pseudomonas putida wcs358 of phoe-caa, a novel hybrid marker gene, which allows monitoring both at the protein level by immunological methods and at the dna level by pcr. the marker is based on the escherichia coli outer membrane protein gene phoe and 75 bp of e. coli caa, which encode a nonbacteriocinic fragment of colicin a. this fragment contains an epitope which is recognized by monoclonal antibody (mab) 1c11. as the epitope is contained ...	1994	7993086
metabolism of chlorofluorocarbons and polybrominated compounds by pseudomonas putida g786(phg-2) via an engineered metabolic pathway.	the recombinant bacterium pseudomonas putida g786(phg-2) metabolizes pentachloroethane to glyoxylate and carbon dioxide, using cytochrome p-450cam and toluene dioxygenase to catalyze consecutive reductive and oxidative dehalogenation reactions (l.p. wackett, m.j. sadowsky, l.n. newman, h.-g. hur, and s. li, nature [london] 368:627-629, 1994). the present study investigated metabolism of brominated and chlorofluorocarbon compounds by the recombinant strain. under anaerobic conditions, p. putida g ...	1994	7993096
loss of the tol meta-cleavage pathway functions of pseudomonas putida strain paw1 (pww0) during growth on toluene.	a derivative of pseudomonas putida strain paw1 bearing the tol plasmid pww0 was isolated from a culture which has grown unlimited on toluene. in contrast to the parent strain paw1, the derivative, strain cg220, is unable to grow with xylenes and toluates, while toluene and benzoate served as substrates. strain cg220 had a remarkable growth advantage against the wild type when grown with toluene. biochemical analysis showed that in strain cg220 toluene was metabolised through the tol plasmid uppe ...	1994	7996396
permeabilization of mycolic-acid-containing actinomycetes for in situ hybridization with fluorescently labelled oligonucleotide probes.	the application of whole-cell hybridization using labelled oligonucleotide probes in microbial systematics and ecology is limited by difficulties in permeabilizing many gram-positive organisms. in this investigation paraformaldehyde treatment, acid methanolysis and acid hydrolysis were evaluated as a means of permeabilizing mycolic-acid-containing actinomycetes prior to hybridization with a fluorescently labelled oligonucleotide probe designed to bind to a conserved sequence of bacterial 16s rrn ...	1994	8000549
enzymatic asymmetric synthesis of alpha-methyl arylalkylamines and alpha-methyl arylalkylalcohols by arylalkyl acylamidases.	with the novel microbial enzyme, 'arylalkyl acylamidase', optically active alpha-methyl arylalkylamines and alpha-methyl arylalkylalcohols have been obtained through enantioselective hydrolysis of their racemic amides and esters. (s)-enantiomers of 1-methylbenzylamine, 1-methyl-3-phenylpropylamine and 1-methyl-3-phenylpropanol of high optical purity (> 94% e.e.) were synthesized with the cells of nocardia erythropolis iam 1440 or cellulomonas fimi aku 671. (r)-enantiomer of 1-methyl-3-phenylprop ...	1994	8000864
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
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
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
reaction engineering aspects of conjugation in biodegradation processes.	conjugation between two pseudomonas putida species has been studied. special emphasis has been given to the design of tools for better defined and reproducible experimental conditions. this is an essential prerequisite for any kinetic analysis of the phenomenon. the experimental approach suggested in the paper is a three-stage continuous fermentation, where donor and recipient strains can be cultivated at predefined steady-state growth rates, and conjugation takes place at steady-state condition ...	1994	8010691
1-amino-2-imidazol-4'-ylethylphosphonic acid is a potent reversible inhibitor of pseudomonas putida histidine ammonia-lyase.	a phosphonic acid analogue of l-histidine, 1-amino-2-imidazol-4'-ylethylphosphonic acid (hisp), was identified as a reversible competitive inhibitor of histidine ammonia-lyase (histidase). the affinity of histidase for hisp was ph dependent, with ki values of 0.28 microm and 10.4 microm compared to substrate km values of 1 and 5 mm at ph 7 and 9, respectively. hisp did not appear to be a substrate for histidase. a twenty-fold molar excess of hisp over enzyme completely protected the active site ...	1994	8012285
transposon-mediated mobilization of chromosomally located catabolic operons of the cam plasmid by tol plasmid transposon tn4652 and cam plasmid transposon tn3614.	the cam (camphor degradation) plasmid is integrated into the chromosome of pseudomonas putida paw-line strains and is not self-transferable as a plasmid via conjugation. our results show that the mobilization of chromosomally located cam and the integration of cam-operons into the chromosome of the new cam+ transconjugants is a reca-independent process mediated by transposons tn4652 (17 kbp) and tn3614 (7.2 kbp). transposon tn3614 is apparently identical to the left-hand and the right-hand seque ...	1994	8012608
siderophore receptor pupa as a marker to monitor wild-type pseudomonas putida wcs358 in natural environments.	for application of genetically engineered fluorescent pseudomonas spp., specific markers are required for monitoring of wild-type pseudomonas strains and their genetically modified derivatives in natural environments. in this study, the specific siderophore receptor pupa of plant growth-promoting pseudomonas putida wcs358 was used as a marker to monitor wild-type strain wcs358. after introduction into natural soil and rhizosphere environments, strain wcs358 could be recovered efficiently on a me ...	1994	8017914
quantitative cell lysis of indigenous microorganisms and rapid extraction of microbial dna from sediment.	this study reports improvements in two of the key steps, lysis of indigenous cells and dna purification, required for achieving a rapid nonselective protocol for extracting nucleic acids directly from sodium dodecyl sulfate (sds)-treated sediment rich in organic matter. incorporation of bead-mill homogenization into the dna extraction procedure doubled the densitometrically determined dna yield (11.8 micrograms of dna.g [dry weight] of sediment-1) relative to incorporation of three cycles of fre ...	1994	8017936
functional and structural relationship of various extradiol aromatic ring-cleavage dioxygenases of pseudomonas origin.	the extradiol ring-cleavage dioxygenases derived from seven different pseudomonas strains were expressed in escherichia coli and the substrate specificities were investigated for a variety of catecholic compounds. the substrate range of four 2,3-dihydroxybiphenyl dioxygenases from biphenyl-utilizing bacteria, 3-methylcatechol dioxygenase from toluene utilizing pseudomonas putida f1, 1,2-dihydroxynaphthalene dioxygenase from a nah7 plasmid, and catechol 2,3-dioxygenase from a tol plasmid pww0 wer ...	1994	8020752
biotransformation of benzothiophene by isopropylbenzene-degrading bacteria.	isopropylbenzene-degrading bacteria, including pseudomonas putida re204, transform benzothiophene to a mixture of compounds. induced strain re204 and a number of its tn5 mutant derivatives were used to accumulate these compounds and their precursors from benzothiophene. these metabolites were subsequently identified by 1h and 13c nuclear magnetic resonance spectroscopy and gas chromatography-mass spectrometry. when strain re204 was incubated with benzothiophene, it produced a bright yellow compo ...	1994	8021182
inability of muconate cycloisomerases to cause dehalogenation during conversion of 2-chloro-cis,cis-muconate.	the conversion of 2-chloro-cis,cis-muconate by muconate cycloisomerase from pseudomonas putida prs2000 yielded two products which by nuclear magnetic resonance spectroscopy were identified as 2-chloro- and 5-chloromuconolactone. high-pressure liquid chromatography analyses showed the same compounds to be formed also by muconate cycloisomerases from acinetobacter calcoaceticus adp1 and pseudomonas sp. strain b13. during 2-chloro-cis,cis-muconate turnover by the enzyme from p. putida, 2-chloromuco ...	1994	8021223
ser-143 is an essential active site residue in histidine ammonia-lyase of pseudomonas putida.	site directed mutagenesis was used to investigate the role of ser-143 in enzyme activity and as a point for attack by cyanide or l-cysteine, two irreversible inhibitors of histidine ammonia-lyase (histidase). two mutant proteins, a s143a substitution and an a142s-s143a transposition, were made. both mutant histidases completely lost all enzymatic activity. western blots with anti-histidase antibodies revealed that the mutant proteins were being expressed at a level equal to that of the wild-type ...	1994	8024588
cloning, heterologous expression, and sequencing of a novel proline iminopeptidase gene, pepi, from lactobacillus delbrueckii subsp. lactis dsm 7290.	the gene for proline iminopeptidase from lactobacillus delbrueckii subsp. lactis dsm 7290 coding for an enzyme that hydrolyses the synthetic substrate l-prolyl-beta-naphthylamide (pro-beta na) was cloned in escherichia coli. an enzymic plate assay was used to screen for positive clones. the gene, designated pepi, was subcloned into vector puc18 and sequenced. the nucleotide sequence revealed an 882 bp open reading frame encoding 294 amino acids, coding for an enzyme with a calculated molecular m ...	1994	8025678
role for the outer membrane ferric siderophore receptor pupb in signal transduction across the bacterial cell envelope.	the outer membrane protein pupb of pseudomonas putida wcs358 facilitates transport of iron complexed to the siderophores pseudobactin bn8 and pseudobactin bn7 into the cell. its synthesis is induced by the presence of these specific siderophores under iron limitation. the signal transduction pathway regulating siderophore-dependent expression of pupb was shown to consist of two regulatory proteins, pupi and pupr, and the pupb receptor itself. mutational analysis of the regulatory genes suggested ...	1994	8026465
purification of a sixth ferredoxin from rhodobacter capsulatus. primary structure and biochemical properties.	a new ferredoxin has been purified from the photosynthetic bacterium rhodobacter capsulatus. it is the sixth ferredoxin to be isolated from this bacterium and it was called fdvi. its primary structure was established based on amino acid sequence analysis of the protein and of peptides derived from it. it is composed of 106 residues including five cysteines. the calculated mass of the polypeptide is 11,402.6 da which matches the experimental value determined by electrospray mass spectrometry. ami ...	1994	8026503
analysis of fluorescent pseudomonads based on 23s ribosomal dna sequences.	the regions from positions 1391 to 1545 and 1620 to 1865 (escherichia coli numbers) of the 23s ribosomal dna sequences have been analyzed for a number of pseudomonas fluorescens and p. putida isolates. variability was observed only in three smaller regions from positions 1484 to 1508, 1531 to 1542, and 1714 to 1748, corresponding to helices 58, 59, and 63, respectively, where up to 53% dissimilarity was found. sequence analysis did not allow clear distinctions among p. fluorescens biovars, p. ch ...	1994	8031104
nucleotide sequence of the gene encoding a repressor for the cytochrome p-450cam hydroxylase operon on the pseudomonas putida cam plasmid.	the camr gene of pseudomonas putida encodes a repressor which regulates expression of the cytochrome p-450cam hydroxylase operon (camdcab). we determined the nucleotide sequence of 1134 continuous base pairs, including the camr gene. when comparing the amino acid sequence deduced from the open reading frame of the gene sequence with that of amino-terminal five residues of the cam repressor, purified from pseudomonas putida, we found that the camr gene encodes a protein of 186 amino acids, with a ...	1994	8031906
purification and characterization of morphinone reductase from pseudomonas putida m10.	the nadh-dependent morphinone reductase from pseudomonas putida m10 catalyses the reduction of morphinone and codeinone to hydromorphone and hydrocodone respectively. morphinone reductase was purified from crude cell extracts to apparent homogeneity in a single affinity-chromatography step using mimetic yellow 2. the purified enzyme was a dimeric flavoprotein with two identical subunits of m(r) 41,100, binding non-covalently one molecule of fmn per subunit. the n-terminal sequence was pdtsfsnpgl ...	1994	8037698
kinetics and thermodynamics of co binding to cytochrome p450nor.	the co-binding reaction of cytochrome p450nor isolated from denitrifying fungus, fusarium oxysporum, has been studied by using a flash photolysis method in the millisecond time domain. we obtained the co on- and off-rate constants in the bimolecular reaction, and determined the activation free energy, enthalpy, and entropy from the temperature dependence of these rate constants. to discuss the structural characteristics of p450nor, these parameters were compared with those of other cytochrome p4 ...	1994	8038156
evidence for the involvement of multiple pathways in the biodegradation of 1- and 2-methylnaphthalene by pseudomonas putida csv86.	pseudomonas putida csv86, a soil bacterium, grows on 1- and 2-methylnaphthalene as the sole source of carbon and energy. in order to deduce the pathways for the biodegradation of 1- and 2-methylnaphthalene, metabolites were isolated from the spent medium and purified by thin layer chromatography. emphasis has been placed on the structural characterisation of isolated intermediates by gc-ms, demonstration of enzyme activities in the cell free extracts and measurement of oxygen uptake by whole cel ...	1994	8042906
significant contribution of arginine-112 and its positive charge of pseudomonas putida cytochrome p-450cam in the electron transport from putidaredoxin.	cytochrome p-450cam of pseudomonas putida is a prototype of various eukaryotic cytochrome p-450 molecules. arg-112 located on the surface of this protein is highly conserved among various other cytochromes p-450. in this study, we constructed mutant genes for p-450cam in which arg-112 was replaced by gln or glu, expressed them in escherichia coli and purified the mutant proteins. their enzymic activities were analyzed in the reconstituted system to determine the function of arg-112. kd values fo ...	1994	8043608
[mossbauer data on the effect of nad.h on the state of iron in bacteria].	the state of fe of bacterial cultures of different systematic positions (bacillus megaterium, bacillus polymyxa, pseudomonas putida, pseudomonas fluorescens, alcaligenes faecalis, arthrobacter siderocapsulatus) grown on the medium containing fe(iii) citrate (up to 100 mg/l) with additional or without nad.h was studied. the samples were in damp air-dry, second moistened, dried at 383 k states. spectra have been obtained at 290 k and 100-200 k. the studied microorganisms have two types of atoms of ...	1994	8043633
[controlled-expression vector for pseudomonas bacteria involving regulatory elements of trpiba genes of pseudomonas putida].	a bireplicon controlled-expression vector pps10 was developed based, on trpiba genes of pseudomonas putida. it is a low-copy-number vector in pseudomonas bacteria, and a high-copy-number vector in escherichia coli. the vector is 10.4 kilobase pairs (kb), determines resistance to kanamycin, carries a replicon of cryptic pseudomonas pmk1 plasmid; a pbr322 replicon; the par locus of pmt2 plasmid; and the trpi gene of p. putida, which encodes the activator protein and the promoter pba of trpba genes ...	1994	8045378
chromosomal gene capture mediated by the pseudomonas putida tol catabolic plasmid.	the pseudomonas putida tol plasmid pww0 is able to mediate chromosomal mobilization in the canonical unidirectional way, i.e., from donor to recipient cells, and bidirectionally, i.e., donor-->recipient-->donor (retrotransfer). transconjugants are recipient cells that have received dna from donor cells, whereas retrotransconjugants are donor bacteria that have received dna from a recipient. the tol plasmid pww0 is able to directly mobilize and retromobilize a kanamycin resistance marker integrat ...	1994	8045894
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
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
tartrate dehydrogenase, a new member of the family of metal-dependent decarboxylating r-hydroxyacid dehydrogenases.	the gene encoding tartrate dehydrogenase has been cloned from pseudomonas putida and sequenced. the gene is 1098 nucleotides long and encodes a protein 365 amino acids in length with a calculated m(r) of 40,636. the gene and the protein encoded by it show strong homology to prokaryotic isopropylmalate dehydrogenases and, to a lesser extent, isocitrate dehydrogenase. thus, tartrate dehydrogenase is the third member to be identified of the family of metal-dependent decarboxylating r-hydroxyacid de ...	1994	8053675
beta-ureidopropionase with n-carbamoyl-alpha-l-amino acid amidohydrolase activity from an aerobic bacterium, pseudomonas putida ifo 12996.	beta-ureidopropionase of aerobic bacterial origin was purified to homogeneity from pseudomonas putida ifo 12996. the enzyme shows a broad substrate specificity. in addition to beta-ureidopropionate (km 3.74 mm, vmax 4.12 u/mg), gamma-ureido-n-butyrate (km 11.6 mm, vmax 19.4 u/mg), and several n-carbamoyl-alpha-amino acids, such as n-carbamoylglycine (km 0.68 mm, vmax 9.14 x 10(-2) u/mg), n-carbamoyl-l-alanine (km 1.56 mm, vmax 1.00 u/mg), n-carbamoyl-l-serine (km 75.1 mm, vmax 3.78 u/mg), and n- ...	1994	8055933
sequence and expression of the todgih genes involved in the last three steps of toluene degradation by pseudomonas putida f1.	the todfc1c2bade gene cluster in pseudomonas putida f1 encodes enzymes for the first four steps of toluene degradation, leading to the formation of 2-hydroxypenta-2,4-dienoate (hpd). here, we report the nucleotide (nt) sequence and expression of the remaining three genes of the tod pathway, downstream from tode and arranged in the order, todgih. the deduced amino acid (aa) sequences of todg [hpd hydratase (268 aa)], todh [4-hydroxy-2-oxovalerate (ho) aldolase (352 aa)] and todi [acylating aldehy ...	1994	8063106
substrate, substrate analogue, and inhibitor interactions with the ferrous active site of catechol 2,3-dioxygenase monitored through xas studies.	the interactions of catechol (substrate), 2-hydroxy-pyridine-n-oxide (substrate analogue), and 2-bromophenol (inhibitor) with the extradiol cleaving catechol-2,3-dioxygenase from pseudomonas putida mt-2 have been monitored through x-ray absorption spectroscopy (xas). the analysis of the data provides details about the mode of coordination of the substrate and of the inhibitors to the active site of the enzyme.	1994	8070565
rhizobia catabolize nod gene-inducing flavonoids via c-ring fission mechanisms.	gas chromatographic and mass spectrometric analyses of derivatized culture medium extracts were used to identify the products of flavonoid metabolism by rhizobia. a number of rhizobium species and biovars degraded their nod gene-inducing flavonoids by mechanisms which originated in a cleavage of the c-ring of the molecule and which yielded conserved a- and b-ring products among the metabolites. in contrast, pseudomonas putida degraded quercetin via an initial fission in its a-ring, and agrobacte ...	1994	8071218
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
cross talk between catabolic pathways in pseudomonas putida: xyls-dependent and -independent activation of the tol meta operon requires the same cis-acting sequences within the pm promoter.	the pm promoter of the meta cleavage operon in the tol (toluene degradation) plasmid pww0 of pseudomonas putida becomes activated by the plasmid-encoded xyls regulator in the presence of benzoate and certain substituted analogs such as 3-methylbenzoate. in the absence of xyls, pm was still responsive to unsubstituted benzoate but with induction kinetics and a range of transcriptional activity which differed substantially from those for the xyls-mediated activation. xyls-independent induction by ...	1994	8071244
trichloroethylene and chloroform degradation by a recombinant pseudomonad expressing soluble methane monooxygenase from methylosinus trichosporium ob3b.	soluble methane monooxygenase (smmo) from methylosinus trichosporium ob3b can degrade many halogenated aliphatic compounds that are found in contaminated soil and groundwater. this enzyme oxidizes the most frequently detected pollutant, trichloroethylene (tce), at least 50 times faster than other enzymes. however, slow growth of the strain, strong competition between tce and methane for smmo, and repression of the smmo locus by low concentrations of copper ions limit the use of this bacterium. t ...	1994	8074526
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
purification and characterization of a novel metal-containing nonheme bromoperoxidase from pseudomonas putida.	a novel bromoperoxidase was purified to homogeneity from the bacterium pseudomonas putida if-3 strain, which produces the antibiotic pyrrolnitrin. the enzyme had a molecular mass of 68,000 and was composed of two identical subunits (33,000). it was specific for i- and br- and inactive toward cl- and f- in the monochlorodimedone assay system. the optimum ph of the enzyme was around 4.2 and it rapidly lost its activity below 3.5, but it was stable over of range ph of 4 to 11. the purified enzyme w ...	1994	8075154
identification of variability of ribosomal dna spacer from pseudomonas soil isolates.	the polymerase chain reaction was used to amplify the spacer region located between the 16s and 23s ribosomal rna genes of strains of pseudomonas fluorescens and pseudomonas putida isolated from peat bog, canola field, or arctic plants. some of spacer region of four of the p. fluorescens strains examined, strains 64-3, 63-28, qp5, and r17-fp2, was about 515 base pairs (bp) in length, and contained the genes for trna(ile) and trna(ala). the dna sequences of two strains from canola, 64-3 and 63-28 ...	1994	8076249
co-regulation by bent dna. functional substitutions of the integration host factor site at sigma 54-dependent promoter pu of the upper-tol operon by intrinsically curved sequences.	the role of integration host factor (ihf) in the regulation of the sigma 54-dependent promoter pu of the tol plasmid of pseudomonas putida has been examined. we have selected in vivo insertions of intrinsically curved dna that restore the responsiveness of an ihf-binding site deletion variant of pu to the cognate activator of the system, xylr. we found five pu derivatives which had inserted a core sequence with 6 phased [a]6 tracts, flanked by different lengths of dna at the location of the form ...	1994	8077217
molecular characterization of 4-hydroxyphenylacetate 3-hydroxylase of escherichia coli. a two-protein component enzyme.	the nucleotide sequences of the hpab and hpac genes encoding the 4-hydroxyphenylacetate 3-hydroxylase from escherichia coli w atcc 11105 have been determined. these genes appear to be part of an operon and encode two proteins of 58,781 and 18,679 da, respectively, that are required for hydroxylase activity. this aromatic hydroxylase is nadh-dependent and uses fad as the redox chromophore. the largest component (hpab) has been purified by affinity chromatography in cibacron blue. e. coli cells th ...	1994	8077235
the vacuolar compartment is required for sulfur amino acid homeostasis in saccharomyces cerevisiae.	in order to isolate new mutations impairing transcriptional regulation of sulfur metabolism in saccharomyces cerevisiae, we used a potent genetic screen based on a gene fusion expressing xyle (from pseudomonas putida) under the control of the promoter region of met25. this selection yielded strains mutated in various different genes. we describe in this paper the properties of one of them, met27. mutation or disruption of met27 leads to a methionine requirement and affects s-adenosylmethionine ( ...	1994	8078479
characterization of the pcar regulatory gene from pseudomonas putida, which is required for the complete degradation of p-hydroxybenzoate.	the pca branch of the beta-ketoadipate pathway in pseudomonas putida is responsible for the complete degradation of p-hydroxybenzoate through ortho cleavage of the initial pathway metabolite, protocatechuate. the pcar regulatory locus has been found to be required for both induction of all of the genes within the pca regulon (pcabdc, pcaij, and pcaf) and the chemotactic response of the bacteria to aromatic compounds. insertional inactivation mutagenesis, using tn5 and mini-tn5 transposons, was u ...	1994	8083169
cometabolic oxidation of polychlorinated biphenyls in soil with a surfactant-based field application vector.	polychlorinated biphenyl (pcb)-degradative genes, under the control of a constitutive promoter, were cloned into a broad-host-range plasmid and a transposon. these constructs were inserted into a surfactant-utilizing strain, pseudomonas putida ipl5, to create a field application vector (fav) in which a surfactant-degrading organism cometabolizes pcb. by utilizing a surfactant not readily available to indigenous populations and a constitutive promoter, selective growth and pcb-degradative gene ex ...	1994	8085825
competition in chemostat culture between pseudomonas strains that use different pathways for the degradation of toluene.	pseudomonas putida mt-2, p. cepacia g4, p. mendocina kr1, and p. putida f1 degrade toluene through different pathways. in this study, we compared the competition behaviors of these strains in chemostat culture at a low growth rate (d = 0.05 h-1), with toluene as the sole source of carbon and energy. either toluene or oxygen was growth limiting. under toluene-limiting conditions, p. mendocina kr1, in which initial attack is by monooxygenation of the aromatic nucleus at the para position, outcompe ...	1994	8085826
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
the induction and repression of benzene and catechol oxidizing capacity of pseudomonas putida ml2 studied in perturbed chemostat culture.	the oxidation of catechol, an intermediate in benzene catabolism, was studied using transient variations in dissolved oxygen tension (dot) when a succinate limited steady state culture of pseudomonas putida ml2 was perturbed with a pulse of another substrate. a model was developed and tested for the effect of fluctuations in oxidizing enzyme activity on dot. it was found that the rate of induction of catechol oxidizing enzymes was independent of dilution rate up to a relative growth rate (mu/mum ...	1994	8085917
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
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
the effect of ferredoxin(bed) overexpression on benzene dioxygenase activity in pseudomonas putida ml2.	the benzene dioxygenase from pseudomonas putida ml2 is a multicomponent complex comprising a flavoprotein reductase, a ferredoxin, and a terminal iron-sulfur protein (isp). the catalytic activity of the isolated complex shows a nonlinear relationship with protein concentration in cell extracts, with the limiting factor for activity in vitro being ferredoxin(bed). the relative levels of the three components were analyzed by using 125i-labelled antibodies, and the functional molar ratio of isp(bed ...	1994	8169199
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
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
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
bacterial morphine dehydrogenase further defines a distinct superfamily of oxidoreductases with diverse functional activities.	pseudomonas putida morphine dehydrogenase is shown to be closely homologous to 18 proteins, defining a superfamily within which morphine dehydrogenase particularly resembles two bacterial, 2,5-dioxo-d-gluconic acid reductases, and two eukaryotic proteins of unknown functions. relationships within the superfamily are extensive and complex. residue identities between protein pairs range from 29-90%. three subgroups are proposed. nevertheless, on the basis of residue conservations/exchanges it is s ...	1994	8192670
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
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
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
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
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
the organization of the pm promoter of the tol plasmid reflects the structure of its cognate activator protein xyls.	the toluate catabolic operon carried by the tol plasmid pww0 of pseudomonas putida is positively regulated by the benzoate-responsive xyls protein which, when activated, stimulates transcription from the operon promoter pm. in this study, the mode in which xyls effects the activity of the pm promoter was examined in vivo by genetic analysis of both protein and promoter variants. substitution of his31asp/ser32pro,leu113pro,phe214leu/glu215a sp/arg216pro or thr312pro, all predicted to disrupt the ...	1994	7969028
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
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
controlled-expression shuttle vector for pseudomonads based on the trpiba genes of pseudomonas putida.	a cloning vector pps7 (8.5 kb) for pseudomonas was constructed from pbr322 and the pseudomonas cryptic low-copy-number pmk1 plasmid. the vector confers resistance to kanamycin (km) and tetracycline (tc), contains the par locus of pseudomonas plasmid pmt2 and a mob site. the new vector was used for construction of controlled-expression vector pps10 (10.4 kb) based on the trpiba genes of pseudomonas putida. this kmr vector contains the trpi gene, encoding activator protein and promoter of trpba ge ...	1994	8125340
analysis of three 2,3-dihydroxybiphenyl 1,2-dioxygenases found in rhodococcus globerulus p6. identification of a new family of extradiol dioxygenases.	the polychlorobiphenyl-degrading bacterium rhodococcus globerulus p6 contains three bphc genes encoding 2,3-dihydroxybiphenyl 1,2-dioxygenases. one of them, bphc1, is clustered with the bphb gene which encodes 2,3-dihydroxy-4-phenylhexa-4,6-diene dehydrogenase and constitutes part of the bph operon specifying the degradation of biphenyl. the nucleotide sequence of bphb and the three bphc genes has been determined. the protein products of the bphbc1 gene cluster were found to exhibit significant ...	1994	8126007
13c nuclear magnetic resonance studies of pseudomonas putida fatty acid metabolic routes involved in poly(3-hydroxyalkanoate) synthesis.	the formation of poly(3-hydroxyalkanoates) (phas) in pseudomonas putida kt2442 from various carbon sources was studied by 13c nuclear magnetic resonance spectroscopy, gas chromatography, and gas chromatography-mass spectroscopy. by using [1-13c]decanoate, the relation between beta-oxidation and pha formation was confirmed. the labeling pattern in phas synthesized from [1-13c]acetate corresponded to the formation of phas via de novo fatty acid biosynthesis. studies with specific inhibitors of the ...	1994	8132461
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