Publications

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
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
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
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
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
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
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
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
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
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
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
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
fpta, the fe(iii)-pyochelin receptor of pseudomonas aeruginosa: a phenolate siderophore receptor homologous to hydroxamate siderophore receptors.	the pseudomonas aeruginosa siderophore pyochelin is structurally unique among siderophores and possesses neither hydroxamate- nor catecholate-chelating groups. the structural gene encoding the 75-kda outer membrane fe(iii)-pyochelin receptor fpta has been isolated by plasmid rescue techniques and sequenced. the n-terminal amino acid sequence of the isolated fpta protein corresponded to that deduced from the nucleotide sequence of the fpta structural gene. the mature fpta protein has 682 amino ac ...	1994	8288523
the role of 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
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
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
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
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
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
fate of pseudomonas putida after release into lake water mesocosms: different survival mechanisms in response to environmental conditions.	to study the fate of pseudomonas putida dsm 3931 in an aquatic environment, cultures of the strain were released into lake water mesocosms. p. putida, bearing the tol-plasmid, was released as a representative xenobiotic-degrading microorganism. the release was carried out in mesocosms with unamended lake water and in lake water with added culture medium to compare the survival of the strain due to the influence of different organic load. as a comparison, the survival of p. putida was followed in ...	1994	24190270
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
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
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
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
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
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
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
a mutagenesis system utilizing a tn1722 derivative containing an escherichia coli-specific vector plasmid: application to pseudomonas species.	a novel transposon (tn) mutagenesis system for gram- non-enteric bacteria was developed which allowed rapid and one-step cloning of the mutated region in escherichia coli. the tn constructed was tn1722-299km, a tn1722 derivative containing a kmr gene and the entire sequence of an e. coli-specific plasmid, pacyc184. the hybrid plasmid consisting of tn1722-299km and the transfer genes of plasmid r388 was conjugally transferred from e. coli to pseudomonas putida or p. aeruginosa, and selection of t ...	1993	8294012
construction of chromosomal reca mutants of pseudomonas putida ppg2.	the reca gene of pseudomonas putida ppg2 was cloned by complementation of the reca mutations of escherichia coli strains dh5 alpha and hb101. the nucleotide sequence of the dna fragment was determined and shown to contain reca and a downstream partial open reading frame. two mutants of p. putida ppg2, strains js387 and js388, were constructed by insertional inactivation of reca with a tetracycline-resistance gene in both orientations. both mutants acquired sensitivity to methyl methanesulfonate ...	1993	8294013
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
indirect utilization of the phytosiderophore mugineic acid as an iron source to rhizosphere fluorescent pseudomonas.	the phytosiderophore mugineic acid (ma) was studied as a source of iron for rhizosphere fluorescent pseudomonads. 55fe supplied as fe-ma was taken up by pseudomonas putida wcs358, b10 and st3 grown under iron deficient conditions. the uptake decreased when the bacteria were grown in the presence of iron. however, no differences in uptake were observed when a siderophore deficient mutant was tested. since ligand exchange between pseudobactin and ma was shown to occur rapidly with a half-life of 2 ...	1993	8358206
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
[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
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
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
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
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
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
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
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
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
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
a new amino acid racemase with threonine alpha-epimerase activity from pseudomonas putida: purification and characterization.	we have found that pseudomonas putida atcc 17642 cells grown in a medium containing d-threonine as the sole nitrogen source produce an enzyme that catalyzes epimerization of threonine. proton nuclear magnetic resonance analysis of the enzyme reaction in deuterium oxide clearly showed epimerization from l- to d-allo-threonine and also from d- to l-allo-threonine. this is the first example of an enzyme that was clearly shown to epimerize threonine. the enzyme has been purified to homogeneity, whic ...	1993	8320235
degradative capability of pseudomonas putida on acetonitrile.	pseudomonas putida, capable of utilizing acetonitrile as a sole source of carbon and nitrogen, was isolated from contaminated soil and water samples collected from industrial sites. the p. putida cells were immobilized in calcium alginate beads. the degradation of acetonitrile by the immobilized cells of p. putida was investigated. the immobilized cells degraded different concentrations of acetonitrile into ammonia and carbon dioxide. the effect of aeration on the degradation rate was also studi ...	1993	8323268
expression of the agga locus of pseudomonas putida in vitro and in planta as detected by the reporter gene, xyle.	in vitro agglutinability by pseudomonas putida, isolate corvallis, with a plant root surface agglutinin is correlated with rapid adhesion of cells of the fluorescent pseudomonad to bean (phaseolus vulgaris) root surfaces. agglutinability in p. putida cells is regulated by nutrient status as well as growth phase. cells grown in three different nutrient complex media are agglutinable at early and mid-late logarithmic phase but become nonagglutinable at stationary phase. cells grown in a minimal me ...	1993	8324250
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
plasmid-mediated mineralization of naphthalene, phenanthrene, and anthracene.	the well-characterized plasmid-encoded naphthalene degradation pathway in pseudomonas putida ppg7(nah7) was used to investigate the role of the nah plasmid-encoded pathway in mineralizing phenanthrene and anthracene. three pseudomonas strains, designated 5r, dfc49, and dfc50, were recovered from a polynuclear aromatic hydrocarbon-degrading inoculum developed from a manufactured gas plant soil slurry reactor. plasmids pka1, pka2, and pka3, approximately 100 kb in size, were isolated from these st ...	1993	8328809
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
complete nucleotide sequence of the 5-exo-hydroxycamphor dehydrogenase gene on the cam plasmid of pseudomonas putida (atcc 17453).	we determined the complete nucleotide sequence of the first gene of pseudomonas putida cytochrome p-450cam hydroxylase operon, camd, which encodes 5-exo-hydroxycamphor dehydrogenase. this dehydrogenase apparently consists of 361 amino acids and has a molecular mass of 38.4 kda. structural relationships to other zinc-containing alcohol dehydrogenases also became evident.	1993	8334169
nicotinoprotein [nad(p)-containing] alcohol/aldehyde oxidoreductases. purification and characterization of a novel type from amycolatopsis methanolica.	extracts of gram-positive bacteria like rhodococcus rhodochrous, rhodococcus erythropolis and amycolatopsis methanolica, but not those of several gram-negative ones, showed dehydrogenase activity for ethanol as well as for methanol when 4-nitroso-n,n-dimethylaniline (ndma) was used as electron acceptor. chromatography of extracts of the first two organisms revealed one activity for both substrates, that of a. methanolica two activities, one of which is able to oxidize methanol and has been purif ...	1993	8385013
energy conservation by pyrroloquinoline quinol-linked xylose oxidation in pseudomonas putida nctc 10936 during carbon-limited growth in chemostat culture.	when grown in carbon source-limited chemostat cultures with lactate or glucose as the carbon and energy source and xylose as an additional source of reducing equivalents. pseudomonas putida nctc 10936 oxidized xylose to xylonolactone and xylonate. no other products were formed from this pentose, nor was it incorporated into biomass. the presence of xylose in these cultures resulted in higher yglucose and ylactate values as compared to cultures without xylose indicating that biologically useful e ...	1993	8385642
in-vivo-generated fusion promoters in pseudomonas putida.	plasmid pest1463 carrying the promoterless pheba operon was cloned into pseudomonas putida paw85, and phenol-utilizing colonies were isolated on minimal plates containing phenol as the only carbon and energy source. in these clones, chromosomally located tn4652 was transposed upstream from the coding sequencing of phea (encoding phenol monooxygenase). sequence analysis together with mapping of the transcription start point of the pheba operon in the recombinant plasmids revealed that fusions of ...	1993	8387446
identification and characterization of the pupb gene encoding an inducible ferric-pseudobactin receptor of pseudomonas putida wcs358.	pseudomonas putida wcs358 can transport iron complexed to a wide variety of pseudobactins produced by other pseudomonas strains. the pupb gene encoding an outer membrane ferric-pseudobactin receptor was isolated from a genomic library of p. putida wcs358. the pupb receptor facilitated iron transport via two distinct heterologous siderophores, i.e. pseudobactin bn8 and pseudobactin bn7. the amino acid sequence deduced from the nucleotide sequence consisted of 804 amino acids (molecular weight 88, ...	1993	8392140
engineering of alkyl- and haloaromatic-responsive gene expression with mini-transposons containing regulated promoters of biodegradative pathways of pseudomonas.	four recombinant mini-tn5 transposons are described which contain outward-facing pm, pu or psal promoters from the catabolic plasmids tol and nah of pseudomonas putida, along with their cognate wild-type regulatory genes (xyls, xylr, nahr) or mutant varieties (xyls2). transcription from such promoters is activated when the host bacteria encounters certain aromatic compounds, such as alkyl- and halobenzoates (xyls, xyls2), alkyl- and halotoluenes (xylr) or salicylates (nahr). these transposons en ...	1993	8393826
essential role of the arg112 residue of cytochrome p450cam for electron transfer from reduced putidaredoxin.	cytochrome p450cam (cyp101) of pseudomonas putida ppg1 in which arg112 is substituted by cys was isolated by in vitro random mutagenesis of the camc gene dna coding for p450cam. the absorption spectra of the purified mutant enzyme were similar to those of the wild type enzyme, but its substrate-dependent nadh oxidation activity in the presence of putidaredoxin (pd) and putidaredoxin reductase (pdr) was extremely low. the rate constant of electron transfer from reduced pd to the heme of the mutan ...	1993	8405387
transformation of 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8- dihydroxyquinoline by quinoline-grown resting cells of pseudomonas putida 86.	resting cells of pseudomonas putida strain 86 were grown on quinoline transformed 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8-dihydroxyquinoline which was not converted further. 7,8-dioxygenating activity was present when the enzymes of quinoline catabolism were induced. quinoline-grown cells of strain 86 treated simultaneously with 2-chloroquinoline and d-(-)-threo-chloramphenicol to prevent protein biosynthesis also formed the cis-7,8-dihydrodiol of 2-chloroquinoline. succinate-grown res ...	1993	8405957
cbbr, a lysr-type transcriptional activator, is required for expression of the autotrophic co2 fixation enzymes of xanthobacter flavus.	xanthobacter flavus is able to grow autotrophically with the enzymes of the calvin cycle for the fixation of co2, which are specified by the cbblsxfp gene cluster. previously, the 5' end of an open reading frame (cbbr), displaying a high sequence similarity to the lysr family of regulatory proteins and transcribed divergently from cbblsxfp, was identified (w. g. meijer, a. c. arnberg, h. g. enequist, p. terpstra, m. e. lidstrom, and l. dijkhuizen, mol. gen. genet. 225:320-330, 1991). this paper ...	1993	8407781
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
a substrate-dependent biological containment system for pseudomonas putida based on the escherichia coli gef gene.	a model substrate-dependent suicide system to biologically contain pseudomonas putida kt2440 is reported. the system consists of two elements. one element carries a fusion between a synthetic lac promoter (pa1-04/03) and the gef gene, which encodes a killing function. this element is contained within a transposaseless mini-tn5 transposon so that it can be integrated at random locations on the pseudomonas chromosome. the second element, harbored by plasmid pcc102, is designed to control the first ...	1993	8285679
use of 5-cyano-2,3-ditolyl tetrazolium chloride for quantifying planktonic and sessile respiring bacteria in drinking water.	direct microscopic quantification of respiring (i.e., viable) bacteria was performed for drinking water samples and biofilms grown on different opaque substrata. water samples or biofilms developed in flowing drinking water were incubated with the vital redox dye 5-cyano-2,3-ditolyl tetrazolium chloride (ctc) and r2a medium. one hour of incubation with 0.5 mm ctc was sufficient to obtain intracellular reduction of ctc to the insoluble fluorescent formazan (ctf) product, which was indicative of c ...	1993	8285688
the preparation of endotoxin-free l-methionine-alpha-deamino-gamma-mercaptomethane-lyase (l-methioninase) from pseudomonas putida.	many types of human and animal tumors have an absolute requirement for methionine. this requirement can be satisfied by homocysteine only in normal cells and tissues. therefore, methionine may be an important target in cancer therapy. to attack this target we have purified endotoxin-free methioninase from pseudomonas putida by a novel and simple procedure. this procedure involves (1) a heat step of the cell extract at 60 degrees c for 8 min, (2) deae-toyopearl ion-exchange chromatography, (3) de ...	1993	8286949
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
growth and plasmid-encoded naphthalene catabolism of pseudomonas putida in batch culture.	the growth characteristics of pseudomonas putida plasmid-harbouring strains which catabolize naphthalene via various pathways in batch culture with naphthalene as the sole source of carbon and energy have been investigated. the strains under study were constructed using the host strain p. putida bs394 which contained various naphthalene degradation plasmids. the highest specific growth rate was ensured by the plasmids that control naphthalene catabolism through the meta-pathway of catechol oxida ...	1993	8270196
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
cloning and expression of a member of a new cytochrome p-450 family: cytochrome p-450lin (cyp111) from pseudomonas incognita.	cytochrome p-450lin catalyzes the 8-methyl hydroxylation of linalool as the first committed step of its utilization by pseudomonas incognita as the sole carbon source. by using a polymerase chain reaction-based cloning strategy, a 2.1-kb dna fragment containing the cytochrome p-450lin gene (linc) was isolated. an open reading frame of 406 amino acids has been identified as that of p-450lin on the basis of amino acid sequence data from peptides of the native protein. heterologous expression of fu ...	1993	8376348
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
nucleotide sequence and initial functional characterization of the clcr gene encoding a lysr family activator of the clcabd chlorocatechol operon in pseudomonas putida.	the 3-chlorocatechol operon clcabd is central to the biodegradative pathway of 3-chlorobenzoate. the clcr regulatory gene, which activates the clcabd operon, was cloned from the region immediately upstream of the operon and was shown to complement an insertion mutation for growth on 3-chlorobenzoate. clcr activated the clca promoter, which controls expression of the clcabd operon, in trans by 14-fold in an in vivo promoter probe assay in pseudomonas putida when cells were incubated with 15 mm 3- ...	1993	8419291
cloning, sequencing, and genetic characterization of regulatory genes, rina and rinb, required for the activation of staphylococcal phage phi 11 int expression.	the int gene of staphylococcal bacteriophage phi 11 is the only viral gene responsible for the integrative recombination of phi 11. to study the regulation of int gene expression, we determined the 5' end of the transcript by s1 mapping. the presumed promoter is located just 22 nucleotides upstream of the int open reading frame in a region which is conserved between phi 11 and a closely related staphylococcal phage, l54a. to clone the possible regulatory gene, a vector which contained the report ...	1993	8432703
proteins induced by sulfate limitation in escherichia coli, pseudomonas putida, or staphylococcus aureus.	two-dimensional gel electrophoresis of proteins from escherichia coli, pseudomonas putida, and staphylococcus aureus, grown with methionine or one of a variety of organosulfates and organosulfonates as the sole source of sulfur, showed expression of specific sets of 7 to 14 proteins which were not observed during growth with sulfate or cysteine for all three species or with thiocyanate for p. putida and s. aureus. under the same conditions, arylsulfatase activity in p. putida and s. aureus was s ...	1993	8432711
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
preelectrophoresis of agarose plugs containing bacterial chromosomal dna prepared for analysis by pulsed field gel electrophoresis can improve the clarity of restriction patterns.	pulsed field gel electrophoresis has indicated that chromosomal dna isolated from stationary phase pseudomonas fluorescens, pseudomonas putida, and escherichia coli cells immobilized in agarose can be fragmented during its release. for p. fluorescens it was demonstrated that the entire chromosome is affected and that there are no specifically fragile sites. the extent of the damage increased both during storage of the dna and also when magnesium ions were provided, suggesting that nucleases may ...	1993	8387734
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
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
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
characterization of cell surface properties in agglutinable and nonagglutinable mutants of pseudomonas putida.	cells of an aggressive, root-colonizing isolate of pseudomonas putida are agglutinated by a root surface glycoprotein. the agglutination phenotype in p. putida isolate corvallis is lacking in mutants (agg-) derived by tn5 insertion and chemical mutagenesis. specific mutation in the agga locus by tn5 insertion results in loss of agglutinability that is complemented in trans by a wild-type copy of the p. putida agga locus. we examined the biochemical bases of agglutination in p. putida by comparin ...	1993	8106135
cosubstrate effects in reductive dehalogenation by pseudomonas putida g786 expressing cytochrome p-450cam.	cytochrome p-450cam was shown to be the primary catalyst mediating reductive dehalogenation of polychlorinated ethanes by pseudomonas putida g786. under anaerobic conditions, the enzyme catalyzed reductive elimination reactions in vivo with the substrates hexachloroethane, pentachloroethane, and 1,1,1,2-tetrachlorethane; the products were tetrachloroethylene, trichloroethylene, and 1,1-dichloroethylene, respectively. in vivo reaction rates were determined. no reaction was observed with 1,1,2,2-t ...	1993	7763853
intrinsic stability and extrinsic stabilization of creatinase from pseudomonas putida.	creatinase (creatine amidinohydrolase, ec 3.5.3.3), a homodimer of 45 kda subunit molecular mass, shows only limited functional stability, and is inaccessible to reconstitution after preceding deactivation, denaturation and dissociation. the enzyme has been characterized regarding its native and denatured states. studying its unfolding characteristics in the presence of "extrinsic factors", such as dte, bsa and glycerol, it was possible to define solvent conditions where the stability of the enz ...	1993	8216893
microbial metabolism of quinoline and related compounds. xix. degradation of 4-methylquinoline and quinoline by pseudomonas putida k1.	a bacterial strain, designated k1, which utilizes 4-methylquinoline and quinoline as sole source of carbon, nitrogen and energy was isolated from soil. based on its morphological and physiological characteristics, it was classified as pseudomonas putida biovar b. four metabolites of 4-methylquinoline degradation were isolated from the culture supernatant and identified as 4-methyl-2-oxo-1,2-dihydroquinoline, 8-hydroxy-4-methyl-2-oxo-1,2-dihydroquinoline, 7,8-dihydroxy-4-methyl-2-oxo-1,2-dihydroq ...	1993	8216899
construction and use of a new broad-host-range lacz transcriptional fusion vector, phrp309, for gram- bacteria.	a new lacz transcriptional fusion vector, phrp309, based on the incq plasmid rsf1010, was constructed and shown to be easily mobilized into a variety of gram- eubacteria. we also developed a two-step cloning procedure to facilitate the cloning of small promoter fragments into the fusion vector. a set of 'cohort' vectors was constructed which allowed directed cloning of fragments downstream from an omega streptomycin/spectinomycin-resistance cassette while maintaining multiple flanking restrictio ...	1993	8224891
metabolism of dibenzothiophene and naphthalene in pseudomonas strains: complete dna sequence of an upper naphthalene catabolic pathway.	from a soil isolate, pseudomonas strain c18, we cloned and sequenced a 9.8-kb dna fragment that encodes dibenzothiophene-degrading enzymes. nine open reading frames were identified and designated doxabdefghij. collectively, we refer to these genes as the dox pathway. at the nucleotide level, doxabd are identical to the ndoabc genes that encode naphthalene dioxygenase of pseudomonas putida. the doxg protein is 97% identical to nahc (1,2-dihydroxynaphthalene dioxygenase) of p. putida. doxe has 37% ...	1993	8226631
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
analysis of the mrna structure of the pseudomonas putida tol meta fission pathway operon around the transcription initiation point, the xylte and the xylfj regions.	the 13 genes encoded by the meta-cleavage operon (approx. 11 kb) of pseudomonas putida tol plasmid pww0 are transcribed from a single promoter, pm. in p. putida, transcription from pm was strictly dependent on the presence of effector-activated xyls protein. three regions of the transcript were analyzed in the wild-type strain of p. putida (pww0) by s1 nuclease protection and primer extension analyses. a major point of transcription initiation was found in the most 5'-end of the operon, which de ...	1993	8241263
early and late responses of tol promoters to pathway inducers: identification of postexponential promoters in pseudomonas putida with lacz-tet bicistronic reporters.	transcriptional lacz fusions to the pu and pm promoters of the tol (toluene degradation) plasmid inserted in monocopy in the chromosome of pseudomonas putida showed a very different responsiveness to their respective aromatic effectors regarding growth phase. while a substantial xyls-dependent activation of pm-lacz was detected nearly instantly after m-toluate addition, xylr- and xylene-mediated induction of the sigma 54 promoter pu became significant only after cells slowed down exponential gro ...	1993	8226632
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
a t7 rna polymerase-based system for the construction of pseudomonas strains with phenotypes dependent on tol-meta pathway effectors.	a general method to construct recombinant pseudomonas putida (and related bacteria), which transcribe specific genes inserted into their chromosome in response to the presence of alkyl- and halobenzoates, has been developed. the system is based on the ability of the t7 rna polymerase (t7pol) to initiate transcription from cognate promoter sequences located upstream from cloned genes. a specialized transposon, mini-tn5 xyls/pm::t7pol, was constructed which contains the structural t7 gene 1 downst ...	1993	8244019
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
essentiality of the three carboxyl-terminal amino acids of the plasmid rk2 replication initiation protein trfa for dna binding and replication activity in gram-negative bacteria.	in a previous study of mutations in trfa, the gene encoding the replication initiation protein of the broad host-range plasmid rk2, a carboxyl-terminal deletion of 3 amino acids of the trfa protein was found to be completely nonfunctional for rk2 replication in escherichia coli and other gram-negative bacteria. in this work site-directed mutagenesis of the trfa gene was carried out to construct trfa proteins altered in the three carboxyl-terminal positions. specifically, trfa proteins with delet ...	1993	8227054
adaptation of pseudomonas putida s12 to high concentrations of styrene and other organic solvents.	pseudomonas putida s12 could adapt to grow on styrene in a two-phase styrene-water system. acetate was toxic for p. putida s12, but cells were similarly able to adapt to higher acetate concentrations. only by using these acetate-adapted cells was growth observed in the presence of supersaturating concentrations of toxic nonmetabolizable solvents such as toluene.	1993	8250572