Publications
| Title | Abstract | Year Filter | PMID(sorted ascending) Filter |
|---|
| nucleotide sequence of the gene for a thermostable esterase from pseudomonas putida mr-2068. | the esterase gene (est) of pseudomonas putida mr-2068 was cloned into escherichia coli jm109. an 8-kb inserted dna directed synthesis of an esterase in e. coli. the esterase gene was in a 1.1-kb psti-clai fragment within the insert dna. the complete nucleotides of the dna fragment containing the esterase gene were sequenced and found to include a single open reading frame of 828 bp coding for a protein of 276 amino acid residues. the open reading frame was confirmed by n-terminal amino acid sequ ... | 1995 | 7670178 |
| polymerase chain reaction amplification of naphthalene-catabolic and 16s rrna gene sequences from indigenous sediment bacteria. | we report the amplification of bacterial genes from uninoculated surface and subsurface sediments by the polymerase chain reaction (pcr). pcr amplification of indigenous bacterial 16s ribosomal dna genes was unsuccessful when subsurface sediment containing approximately 10(7) cells.g-1 was added directly to a pcr mixture. however, when 10 mg of sediment was inoculated with approximately 10(5) cells of pseudomonas putida g7, the nahac naphthalene dioxygenase gene characteristic of the p. putida g ... | 1993 | 7683182 |
| human glyoxalase i. cdna cloning, expression, and sequence similarity to glyoxalase i from pseudomonas putida. | glyoxalase i (ec 4.4.1.5) catalyzes the transformation of methylglyoxal and glutathione to s-lactoylglutathione. we have isolated human cdna clones encoding glyoxalase i from a phorbol myristate acetate-treated u937 cdna library. this cdna encodes a protein of 184 amino acids with a calculated m(r) of 20,719. the amino acid composition calculated from the deduced amino acid sequence agreed with that reported for glyoxalase i purified from human erythrocytes. the escherichia coli cells carrying t ... | 1993 | 7684374 |
| biological synthesis of the analgesic hydromorphone, an intermediate in the metabolism of morphine, by pseudomonas putida m10. | the morphine alkaloid hydromorphone (dihydromorphinone) was identified as an intermediary metabolite in the degradation of morphine by pseudomonas putida m10. a constitutive nadh-dependent morphinone reductase capable of catalyzing the reduction of the 7,8-unsaturated bond of morphinone and codeinone, yielding hydromorphone and hydrocodone, respectively, was shown to be present in cell extracts. the structures have been identified by 1h nuclear magnetic resonance and mass spectrometry. morphinon ... | 1993 | 7689317 |
| cloning and sequencing of a dehalogenase gene encoding an enzyme with hydrolase activity involved in the degradation of gamma-hexachlorocyclohexane in pseudomonas paucimobilis. | in pseudomonas paucimobilis ut26, gamma-hexachlorocyclohexane (gamma-hch) is converted by two steps of dehydrochlorination to a chemically unstable intermediate, 1,3,4,6-tetrachloro-1,4-cyclohexadiene (1,4-tcdn), which is then metabolized to 2,5-dichloro-2,5-cyclohexadiene-1,4-diol (2,5-ddol) by two steps of hydrolytic dehalogenation via the chemically unstable intermediate 2,4,5-trichloro-2,5-cyclohexadiene-1-ol (2,4,5-dnol). to clone a gene encoding the enzyme responsible for the conversion of ... | 1993 | 7691794 |
| transcription of the cam operon and camr genes in pseudomonas putida ppg1. | in pseudomonas putida carrying the cam plasmid, the operon (camdcab) encoding enzymes involved in the degradation pathway of d-camphor is negatively regulated by the camr protein, and camr is autorepressed. s1 nuclease mapping revealed that camdcab and camr were divergently transcribed from overlapping promoters, the transcription start sites were separated by 11 bp, and transcriptions of the cam operon (camdcab) and camr increased about 10- and 4-fold, respectively, immediately after addition o ... | 1993 | 7693653 |
| alginate regulatory and biosynthetic gene homologs in pseudomonas putida wcs358: correlation with the siderophore regulatory gene pfra. | a previous study [venturi et al., mol. microbiol. 10 (1993) 63-73] demonstrated that the siderophore regulatory gene pfra of pseudomonas putida (pp) wcs358 is highly similar and interchangeable with the alginate regulatory gene algq (algr2) of p. aeruginosa (pa). the algq gene is physically linked to two other alginate regulators in the pa chromosome, namely algr (algr1), a response regulator, and algp (algr3), a histone-like gene. in this study, we have identified the same genes and a similar g ... | 1995 | 7698672 |
| cloning and sequencing of a 29 kb region of the bacillus subtilis genome containing the hut and wapa loci. | within the framework of an international project for the sequencing of the entire bacillus subtilis genome, a 29 kb chromosome segment, which contains the hut operon (335 degrees) and the wapa gene, has been cloned and sequenced. this region (28,954 bp) contains 21 complete orfs and one partial one. the 5th, 6th and 17th genes correspond to huth encoding histidase, hutp encoding the positive regulator for the hut operon and wapa encoding a precursor of three major wall-associated proteins, respe ... | 1995 | 7704263 |
| degradation of 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 |
| cryptic dehalogenase and chloroamidase genes in pseudomonas putida and the influence of environmental conditions on their expression. | mutants of two strains of pseudomonas putida expressed two cryptic chloroamidases (c-amidase and h-amidase) and one cryptic dehalogenase (dehii). the mutants were selected on either 2-chloropropionamide (2cpa) or 2-monochloropropionate (2mcpa), developing as papillae in parental colonies growing on a metabolisable support substrate. mutants expressing c-amidase were selected if 2cpa was utilised as either a carbon or a nitrogen source. h-amidase mutants were selected only if 2cpa was used as a n ... | 1995 | 7710321 |
| tetrameric structure and cellular location of catechol 2,3-dioxygenase. | catechol 2,3-dioxygenase from the meta-cleavage pathway encoded on the tol plasmid of pseudomonas putida (pwwo) was investigated by electron microscopy. negatively stained samples of the purified catechol 2,3-dioxygenase revealed that the enzyme consists of four subunits arranged in a tetrahedral conformation. monoclonal antibodies raised against catechol 2,3-dioxygenase showed highly specific reactions and were used to localize the enzyme in escherichia coli (paw31) and p. putida (pwwo), using ... | 1995 | 7710322 |
| effect of cellular physiology on pcr amplification efficiency. | culture conditions, and other variables that modulate a cell's physiology, can bias a polymerase chain reaction (pcr) amplification against generating a representative population profile. two pseudomonas putida nahr alleles were constructed in puc19 that differ solely in a 31-bp internal segment whose sequence has been inverted. after pcr amplification, the products could be distinguished on the basis of a change in a unique restriction site. when an escherichia coli strain carrying one nahr all ... | 1995 | 7711949 |
| a two-protein component 7 alpha-cephem-methoxylase encoded by two genes of the cephamycin c cluster converts cephalosporin c to 7-methoxycephalosporin c. | two genes, cmci and cmcj, corresponding to open reading frames 7 and 8 (orf7 and orf8) of the cephamycin c cluster of nocardia lactamdurans encode enzymes that convert cephalosporin c to 7-methoxycephalosporin c. proteins p7 and p8 (the products of orf7 and orf8 expressed in streptomyces lividans) introduce the methoxyl group at c-7 of the cephem nucleus. efficient hydroxylation at c-7 and transfer of the methyl group from s-adenosylmethionine require both proteins p7 and p8, although p7 alone s ... | 1995 | 7721717 |
| comparative ribosomal protein sequence analyses of a phylogenetically defined genus, pseudomonas, and its relatives. | i analyzed various families of ribosomal proteins obtained from selected species belonging to the genus pseudomonas sensu stricto and allied organisms which were previously classified in the genus pseudomonas. partial amino acid sequencing of l30 preparations revealed that the strains which i examined could be divided into three clusters. the first cluster, which was assigned to the genus pseudomonas sensu stricto, included pseudomonas aeruginosa, pseudomonas putida, pseudomonas mendocina, and p ... | 1995 | 7727274 |
| three distinct quinoprotein alcohol dehydrogenases are expressed when pseudomonas putida is grown on different alcohols. | a bacterial strain that can utilize several kinds of alcohols as its sole carbon and energy sources was isolated from soil and tentatively identified as pseudomonas putida hk5. three distinct dye-linked alcohol dehydrogenases (adhs), each of which contained the prosthetic group pyrroloquinoline quinone (pqq), were formed in the soluble fractions of this strain grown on different alcohols. adh i was formed most abundantly in the cells grown on ethanol and was similar to the quinoprotein adh repor ... | 1995 | 7730276 |
| identification of active site residues by site-directed mutagenesis of delta 5-3-ketosteroid isomerase from pseudomonas putida biotype b. | in order to assess the roles of specific amino acid residues in the delta 5-3-ketosteroid isomerase from pseudomonas putida biotype b during catalysis, we replaced aspartic acid 40 with asparagine (d40n) and tyrosine 16 with phenylalanine (y16f) in the enzyme by site-directed mutagenesis. both purified mutant enzymes resulted in profound decreases in catalytic activities, 10(3.3)-fold in the y16f mutant and 10(6.2)-fold in the d40n mutant. aspartic acid 40 and tyrosine 16 of the enzyme are the c ... | 1995 | 7730300 |
| the evolutionary relationship of biphenyl dioxygenase from gram-positive rhodococcus globerulus p6 to multicomponent dioxygenases from gram-negative bacteria. | the gram+ bacterium rhodococcus globerulus p6 (rgp6) catabolizes a range of polychlorinated biphenyl (pcb) congeners, thus being of interest in bioelimination processes for pcb. the first step in the pathway, a dioxygenase attack of one of the biphenyl (bp) rings, is catalyzed by biphenyl dioxygenase (bdo). in this study, the nucleotide (nt) sequences of the four clustered cistrons, bpha1a2a3a4, encoding the subunits of bdo and forming part of the bph operon of rgp6 for bp degradation, were dete ... | 1995 | 7737502 |
| substrate specificity differences between two catechol 2,3-dioxygenases encoded by the tol and nah plasmids from pseudomonas putida. | the substrate specificities of two catechol 2,3-dioxygenases, one encoded by xyle on the tol plasmid pww0 and the other encoded by nahh on the nah7 plasmid, were investigated. the xyle catechol 2,3-dioxygenase catalyzes the ring-cleavage of catechol, 3-methylcatechol and 4-methylcatechol. the nahh catechol 2,3-dioxygenase was partially deficient in oxidizing 3-methylcatechol due to defects in two catalytic properties. first, nahh has a lower kcat value for 3-methylcatechol compared to xyle, and ... | 1995 | 7744021 |
| purification and characterization of an atp-dependent amidohydrolase, n-methylhydantoin amidohydrolase, from pseudomonas putida 77. | n-methylhydantoin amidohydrolase, an atp-dependent amidohydrolase involved in microbial degradation of creatinine, was purified 70-fold to homogeneity, with a 62% overall recovery, and was crystallized from pseudomonas putida 77. the enzyme has a relative molecular mass of 300,000. it is a tetramer of two identical small subunits (m(r) 70,000) and two identical large subunits (m(r) 80,000). the enzyme requires atp for the amidohydrolysis of n-methylhydantoin and vice versa. mg2+, mn2+ or co2+, a ... | 1995 | 7744042 |
| iron-responsive gene expression in pseudomonas fluorescens m114: cloning and characterization of a transcription-activating factor, pbra. | in response to iron limitation. pseudomonas fluorescens m114 induces a number of genes including an iron-scavenging siderophore termed pseudobactin m114, its cognate receptor, pbua, and a casein protease. a tn5lacz-induced mutant (m114fa1) was isolated that exhibits a pleiotropic phenotype and lacks the ability to express these iron-regulated genes. a cosmid clone was identified which complements this mutation. this clone is capable of activating a number of iron-regulated promoter fusion constr ... | 1995 | 7746151 |
| [degradation of syntanol ds-10 by bacteria immobilized in polysaccharide gels]. | degradative capability of bacterium pseudomonas putida immobilized in polysaccharide (agar-agar or furcellarane) gels towards a nonionic surfactant sintanol ds-10 was studied. the immobilized cells were shown to retain their ability to degrade the substrate in the model experiments as well as in the real waste water. the substrate was degraded after adsorption to the carrier beads. | 1995 | 7746826 |
| a homolog of the rhizobium meliloti nitrogen fixation gene fixn is involved in the production of a microaerobically induced oxidase activity in the phytopathogenic bacterium agrobacterium tumefaciens. | hybridization analysis using the rhizobium meliloti nitrogen fixation gene fixn as a probe revealed the presence of a homologous dna region in the phytopathogenic bacterium agrobacterium tumefaciens. hybridization signals were also detected with total dnas of rhizobium leguminosarum bv. phaseoli, rhodobacter capsulatus and escherichia coli, but not those of xanthomonas campestris pv. campestris and pseudomonas putida. the hybridizing fragment from a. tumefaciens was cloned and sequenced. the pre ... | 1995 | 7753030 |
| localization and organization of phenol degradation genes of pseudomonas putida strain h. | the genetic organization of the dna region encoding the phenol degradation pathway of pseudomonas putida h has been investigated. this strain can utilize phenol or some of its methylated derivatives as its sole source of carbon and energy. the first step in this process is the conversion of phenol into catechol. catechol is then further metabolized via the meta-cleavage pathway into tca cycle intermediates. genes encoding these enzymes are clustered on the plasmid ppgh1. a region of contiguous d ... | 1995 | 7753034 |
| x-ray absorption spectroscopic studies of the fe(ii) active site of catechol 2,3-dioxygenase. implications for the extradiol cleavage mechanism. | the extradiol-cleaving catechol 2,3-dioxygenase (2,3-ctd) isolated from pseudomonas putida mt-2 and its catechol and ternary e.s.no complexes are characterized by x-ray absorption spectroscopy (xas). the intensities of the 1s-->3d transitions in the pre-edge spectra of the uncomplexed enzyme and its substrate complex show that the fe(ii) center is five-coordinate in both complexes, in agreement with earlier magnetic circular dichroism studies [mabrouk, p. a., orville, a. m., lipscomb, j. d., & s ... | 1995 | 7756296 |
| random walk calculations for bacterial migration in porous media. | bacterial migration is important in understanding many practical problems ranging from disease pathogenesis to the bioremediation of hazardous waste in the environment. our laboratory has been successful in quantifying bacterial migration in fluid media through experiment and the use of population balance equations and cellular level simulations that incorporate parameters based on a fundamental description of the microscopic motion of bacteria. the present work is part of an effort to extend th ... | 1995 | 7756547 |
| [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 |
| cytological aspects of resistance to potassium tellurite conferred on pseudomonas cells by plasmids. | the ultrastructure of strains pseudomonas putida bs228 and pseudomonas aeruginosa ml4262 harboring plasmids pbs10, pbs31, and pbs221, which determine resistance to potassium tellurite, was studied. bacteria were grown in media containing increasing concentrations of potassium tellurite. crystalline structures containing tellurium appeared in their periplasmic space. the dynamics of crystal growth was studied. crystals were released into the medium by pinching off of the outer membrane vesicles c ... | 1995 | 7763135 |
| 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 |
| aerobic, phenol-induced tce degradation in completely mixed, continuous-culture reactors. | both pseudomonas putida f1 and a mixed culture were used to study tce degradation in continuous culture under aerobic, non-methanotrophic conditions. tce mass balance studies were performed with continuous culture reactors to determine the total percent removed in the reactors, and to quantify the percent removed by air stripping and biodegradation. adsorption of tce to biomass was assumed to be negligible. this research demonstrated the feasibility of treating tce-contaminated water under aerob ... | 1993 | 7763855 |
| survival and impact of genetically engineered pseudomonas putida harboring mercury resistance gene in aquatic microcosms. | the survival of wild-type and genetically engineered pseudomonas putida ppy101 that contained a recombinant plasmid psr134 conferring mercury resistance were monitored in aquatic microcosms. we used lake, river, and spring water samples. the density of genetically engineered and wild-type p. putida decreased rapidly within 5 days (population change rate k -0.87 approximately -1.00 day-1), then moderately after 5 to 28 days (-0.10 approximately -0.14 day-1). the population change rates of genetic ... | 1993 | 7764012 |
| survival and impact of genetically engineered pseudomonas putida harboring mercury resistance gene in soil microcosms. | the survival of genetically engineered and wild-type pseudomonas putida ppy101, that contained a recombinant plasmid psr134 conferring mercury resistance, were monitored in andosol and sand microcosms. the survival of genetically engineered and wild-type p. putida was not significantly different in andosol. the population change of the two strains was dissimilar in andosol and sand. the survival of genetically engineered and wild-type p. putida strains was affected by the water content of andoso ... | 1994 | 7764510 |
| cloning and sequence analysis of a plasmid-encoded 2-haloacid dehalogenase gene from pseudomonas putida no. 109. | the 2-haloacid dehalogenase of pseudomonas putida no. 109 was mediated by a 74-kb conjugative plasmid, which was transferred by mating into pseudomonas and escherichia coli and there expressed the dehalogenase. a 2.8-kb ecori-fragment generated from the plasmid was cloned and sequenced. the dehalogenase gene (dehh109) was identified by comparison with the n-terminal amino acid sequence and the molecular weight of the enzyme protein. the gene dehh109 coded for a 224-amino acid protein of m(r) 25, ... | 1994 | 7764511 |
| transformation of pseudomonas putida by electroporation. | the optimum electrotransformation conditions were determined for pseudomonas putida ppy101 with plasmid psup104 (9.5 kb) and psr134 (18.6 kb). field strength was a very important parameter for electrotransformation efficiency. optimum efficiencies (1.1 x 10(5) transformants/micrograms dna) with psup104 and psr134 were obtained at a field strength of 12.5 kv/cm, a time constant of about 4.5 ms (resistance setting of 200 ohms), a supercoiled dna concentration of 100 ng/ml, and a cell concentration ... | 1994 | 7764975 |
| tol plasmid-specified xylene oxygenase is a wide substrate range monooxygenase capable of olefin epoxidation. | xylene oxygenase, which is encoded on the tol plasmid pwwo of pseudomonas putida mt-2, is a key enzyme system in the degradation of toluene and xylenes by this organism. it was expressed in an escherichia coli recombinant strain carrying the xylma structural genes. this recombinant, which expressed xylene oxygenase from the heat-shock induced lambda pl promoter, was analyzed for its potential as a biocatalytic tool so as to effect the oxidation of side chains of aromatic hydrocarbons to the corr ... | 1994 | 7764991 |
| production, partial characterization, and potential diagnostic use of salicylate hydroxylase from pseudomonas putida uuc-1. | an unusual strain of pseudomonas putida uuc-1 capable of growth at high salicylate concentration (10 gl-1) was investigated with the aim of developing an assay and a biosensor system for determining salicylate in body fluids by utilizing the salicylate hydroxylase enzyme. medium and growth condition optimization were carried out under chemostat conditions. the highest biomass yield was at 4.0 gl-1 salicylate, 25 degrees c, ph 6.5, and 0.2 h-1 dilution rate. growth occurred at up to 0.45 h-1 dilu ... | 1994 | 7765077 |
| characterization of phe b gene encoding catechol 2,3-dioxygenase. | dna sequence of the phe b gene isolated from a chromosome of the phenol degrading bacterium pseudomonas putida bh is identical to that of the dmp b gene from the phenol degradative plasmid, pvi150. catechol 2,3-dioxygenase encoded by phe b showed similar substrate specificity to that of xyl e. however, phe b has much smaller km values than xyl e, indicating that phe b is useful for treatment of low concentrations of catechol derivatives in waste water. | 1994 | 7765392 |
| cloning and expression of pseudomonas putida esterase gene in escherichia coli and its use in enzymatic production of d-beta-acetylthioisobutyric acid. | the esterase catalyzes the stereoselective hydrolysis of methyl dl-beta-acetylthioisobutyrate (dl-ester) to give d-beta-acetylthioisobutyric acid (dat). to use the enzyme for dat production, the esterase gene of pseudomonas putida was cloned and expressed in e. coli. the recombinant e. coli containing the esterase gene produced a large amount of the enzyme in an active form. this strain could be used for the asymmetric synthesis of dat. | 1994 | 7765492 |
| cloning, sequence analysis, and expression of the gene encoding formaldehyde dismutase from pseudomonas putida f61. | the gene (fdm) coding for formaldehyde dismutase (ec 1.2.99.4) from a genomic library of formaldehyde-tolerant pseudomonas putida f61 was cloned and expressed in escherichia coli. the nucleotides of the cloned dna were sequenced; they included a single open reading frame of 1200 base pairs, coding for a putative protein with a molecular weight of 42,848. sequencing of the first 20 n-terminal amino acid residues and of an internal part of the enzyme purified from p. putida f61 established the ide ... | 1995 | 7766017 |
| use of a polymerase-chain-reaction-amplified dna probe from pseudomonas putida to detect d-hydantoinase-producing microorganisms by direct colony hybridization. | pseudomonas putida strain dsm 84 produces n-carbamyl-d-amino acids from the corresponding d-5-monosubstituted hydantoins. the sequence of the d-hydantoinase gene from this strain (genbank accession number l24157) was used to develop a dna probe of 122 base pairs (bp) that could detect d-hydantoinase genes in other bacterial genera by dna and by colony hybridization. under conditions tolerating 32% mismatch, the probe was specific for all strains that expressed d-hydantoinase activity. these incl ... | 1995 | 7766091 |
| biodegradation of 2-chloroethanol by freely suspended and adsorbed immobilized pseudomonas putida us2 in soil. | the degradation of 2-chloroethanol by pseudomonas putida us2 was investigated in batch, repeated batch and continuous cultures especially in a packed-bed fermenter with sand. the degradation of 2-chloroethanol was connected with a release of protons, which led to a decrease of the ph in the medium. higher initial concentration than 25 mm 2-chloroethanol were not degraded completely because they entailed a decrease of the ph to 5.0, which inhibited further growth and degradation. p. putida us2 sh ... | 1995 | 7766127 |
| purification and characterization of a cam repressor (camr) for the cytochrome p-450cam hydroxylase operon on the pseudomonas putida cam plasmid. | the cytochrome p-450cam hydroxylase operon of pseudomonas putida ppg1 (atcc 17543) encodes proteins responsible for early steps of the degradation of d-camphor. transcription of this operon is negatively controlled by the cam repressor (camr), and the expression of camr is autoregulated. camr was purified from escherichia coli harboring an overproducing plasmid. the repressor forms a homodimer with a molecular mass of 40 kda, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis ... | 1995 | 7768809 |
| involvement of ihf protein in expression of the ps promoter of the pseudomonas putida tol plasmid. | regulation of the xyl gene operons of the pseudomonas putida tol plasmid is mediated by the products of the downstream clustered and divergently oriented xylr and xyls regulatory genes. the xylr-xyls intergenic region contains the xylr and xyls promoters pr and ps, respectively. a binding site for the xylr activator protein is located upstream of ps and overlapping pr. dnase i footprint experiments showed that one of these sites, which overlaps the recognition site for xylr activator, as well as ... | 1995 | 7768832 |
| evidence for a (triosephosphate isomerase-like) "catalytic loop" near the active site of glyoxalase i. | the conformational mobility of glyoxalase i (glx i) during catalysis has been probed using stable analogs of the enediol intermediate that forms along the reaction pathway: gsc(o)n(oh)r, where gs = glutathionyl and r = ch3 (1), c6h5 (2), c6h4cl (3), or c6h4br (4). for human erythrocyte glx i, catalysis is unlikely to be coupled to major changes in protein secondary structure, as the circular dichroism spectrum of the enzyme (190-260 nm) is insensitive to saturating concentrations of either enedi ... | 1995 | 7768882 |
| effect of cultivation medium on some physicochemical parameters of outer bacterial membrane. | the changes of surface charge and hydrophobicity of the outer bacterial membrane in relation to utilization of n-hexadecane were studied. for this spectrophotometric study adsorption of methylene blue and transport of gentian violet were used. the decrease in the negative charge of the bacterial strains pseudomonas putida ccm 3423, p. aeruginosa, and p. fluorescens ccm 2115, depended on the type of growth medium. the decrease of surface charge was in the order: meat extract peptone broth > miner ... | 1995 | 7770007 |
| nucleotide sequence of the staphylococcus aureus rna polymerase rpob gene and comparison of its predicted amino acid sequence with those of other bacteria. | the complete nucleotide sequence of the rpob gene which encodes the beta subunit of s. aureus rna polymerase has been determined. the rpob protein, consists of 1182 amino acids and has a novel initiation codon uug which initiates protein synthesis with methionine. there is a very strong shine-dalgarno complementarity and the -10 and -35 promoter hexameric sequences are taatat and ccgttt, respectively. a rho-dependent termination site, caatcaa, is present which overlaps the -35 promoter sequence ... | 1995 | 7772603 |
| isolation and characterisation of a strain of pseudomonas putida that can express a periplasmic nitrate reductase. | a strain of pseudomonas putida that can express a nitrate reductase that is located in the periplasmic compartment was isolated from freshwater. the enzyme was active in vivo during arginine fermentation and at the onset of oxygen limitation in batch cultures. the activity of the enzyme increased the yield of bacteria following fermentative growth under anoxic conditions with arginine, but nitrate reduction did not support growth on non-fermentable carbon substrates under anoxic conditions. cell ... | 1995 | 7778973 |
| pcr amplification and direct sequencing of gyrb genes with universal primers and their application to the detection and taxonomic analysis of pseudomonas putida strains. | degenerate pcr primers, up-1 and up-2r, for the amplification of dna gyrase subunit b genes (gyrb) were designed by using consensus amino acid sequences of gyrases from escherichia coli, pseudomonas putida, and bacillus subtilis. in addition to the degenerate sequences, these primers have sequences at the 5' end which allow direct sequencing of amplified pcr products. with these primers, dna segments of the predicted size were amplified from a variety of gram-negative and gram-positive genera. t ... | 1995 | 7793912 |
| characterization of biphenyl catabolic genes of gram-positive polychlorinated biphenyl degrader rhodococcus sp. strain rha1. | rhodococcus sp. strain rha1 is a gram-positive polychlorinated biphenyl (pcb) degrader which can degrade 10 ppm of pcb48 (equivalent to aroclor1248), including tri-, tetra-, and pentachlorobiphenyls, in a few days. we isolated the 7.6-kb ecori-bamhi fragment carrying the biphenyl catabolic genes of rha1 and determined their nucleotide sequence. on the basis of deduced amino acid sequence homology, we identified six bph genes, bpha1a2a3a4, bphb, and bphc, that are responsible for the initial thre ... | 1995 | 7793929 |
| combination of the tod and the tol pathways in redesigning a metabolic route of pseudomonas putida for the mineralization of a benzene, toluene, and p-xylene mixture. | construction of a hybrid strain which is capable of mineralizing components of a benzene, toluene, and p-xylene mixture simultaneously was attempted by redesigning the metabolic pathway of pseudomonas putida. genetic and biochemical analyses of the tod and the tol pathways revealed that dihydrodiols formed from benzene, toluene, and p-xylene by toluene dioxygenase in the tod pathway could be channeled into the tol pathway by the action of cis-p-toluate-dihydrodiol dehydrogenase, leading to compl ... | 1995 | 7793941 |
| cloning of salicylate hydroxylase gene and catechol 2,3-dioxygenase gene and sequencing of an intergenic sequence between the two genes of pseudomonas putida kf715. | the salicylate hydroxylase can convert the salicylate to catechol, and catechol 2,3-dioxygenase catalizes the conversion of catechol to 2-hydroxymuconic semialdehyde. a salicylate hydroxylase gene and a catechol 2,3-dioxygenase have been cloned from chromosomal dna of p. putida kf715. the two genes have different promoters. an open reading frame with 339 nucleotides preceded by a putative ribosome-binding sequence (ggagg) was identified in the intergenic sequence between salicylate hydroxylase g ... | 1995 | 7794247 |
| site-specific deletions of chromosomally located dna segments with the multimer resolution system of broad-host-range plasmid rp4. | the multimer resolution system (mrs) of the broad-host-range plasmid rp4 has been exploited to develop a general method that permits the precise excision of chromosomal segments in a variety of gram-negative bacteria. the procedure is based on the site-specific recombination between two directly repeated 140-bp resolution (res) sequences of rp4 effected by the plasmid-borne resolvase encoded by the para gene. the efficiency and accuracy of the mrs system to delete portions of chromosomal dna fla ... | 1995 | 7798149 |
| interaction of two lysr-type regulatory proteins catr and clcr with heterologous promoters: functional and evolutionary implications. | the soil bacteria pseudomonas putida can use benzoate or 3-chlorobenzoate as a sole carbon source. benzoate and 3-chlorobenzoate are converted into catechol and 3-chlorocatechol, respectively, which are in turn converted into tricarboxylic acid cycle intermediates. the catabolic pathways of both compounds proceed through similar intermediates, have similar genetic organization, and have homologous enzymes responsible for different catabolic steps. this has led to suggestions that the plasmid-bor ... | 1994 | 7809047 |
| adaptation of pseudomonas putida s12 to ethanol and toluene at the level of fatty acid composition of membranes. | pseudomonas putida s12 was more tolerant to ethanol when preadapted to supersaturating concentrations of toluene. cellular reactions at the membrane level to the toxicities of both compounds were different. in growing cells of p. putida s12, sublethal concentrations of toluene resulted in an increase in the degree of saturation of the membrane fatty acids, whereas toxically equivalent concentrations of ethanol led to a decrease in this value. contrary to this, cells also reacted to both substanc ... | 1994 | 7811084 |
| 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 |
| discontinuities in the evolution of pseudomonas putida cat genes. | the organization and transcriptional control of chromosomal cat genes (required for dissimilation of catechol by the beta-ketoadipate pathway) in the pseudomonas putida biotype strain (atcc 12633) are reported. nucleotide sequence reveals that catr is separated by 135 bp from the divergently transcribed catbc,a; catc begins 21 nucleotides downstream from catb, and cata begins 41 nucleotides downstream from catc. this contrasts with the gene arrangement in other bacteria, in which cata lies sever ... | 1995 | 7814330 |
| modeling cytochrome p450 14 alpha demethylase (candida albicans) from p450cam. | the tertiary structure of cytochrome p450 14 alpha demethylase--candida albicans (p450 ca) is modeled on the basis of sequence alignment with two closely related proteins and the crystallographic structure of pseudomonas putida p450cam. the secondary structure prediction system used combines the information from several algorithms and trains the data to offer an optimized prediction of the known p450cam. the trained algorithm was then used to predict the secondary structure of the other p450 seq ... | 1994 | 7819160 |
| cloning and sequences of the first eight genes of the chromosomally encoded (methyl) phenol degradation pathway from pseudomonas putida p35x. | pseudomonas putida p35x (ncib 9869) metabolises phenol and cresols via a chromosomally encoded meta-cleavage pathway. a 13.4-kb fragment of the chromosome involved in encoding phenol catabolism was cloned and characterized. deletion analysis and nucleotide sequencing of a 6589-bp region, in conjunction with enzyme assays, were used to identify the phhklmnop genes encoding the phenol hydroxylase, the phhb gene encoding catechol 2,3-dioxygenase (ec 1.13.11.2) and the phhq gene that encodes a small ... | 1994 | 7828892 |
| cloning cassettes containing the reporter gene xyle. | two puc-derived vectors containing the promoterless xyle gene (encoding catechol 2,3-dioxygenase) of pseudomonas putida mt-2 were constructed. the t(o) transcriptional terminator of phage lambda was placed downstream from the stop codon of xyle. the new vectors, pxt1 and pxt2, contain xyle and the t(o) terminator within a cloning cassette which can be excised with several endonucleases. when inserted into a transcribed sequence, this xyle cassette reports promoter activity and interrupts downstr ... | 1994 | 7828900 |
| localization of functional domains in the escherichia coli coprogen receptor fhue and the pseudomonas putida ferric-pseudobactin 358 receptor pupa. | transport of ferric-siderophores across the outer membrane of gram-negative bacteria is mediated by specific outer membrane receptors. to localize the substrate-binding domain of the ferric-pseudobactin 358 receptor, pupa, of pseudomonas putida wcs358, we constructed chimeric receptors in which different domains of pupa were replaced by the corresponding domains of the related ferric-pseudobactin receptors pupb and pupx, or the coprogen receptor fhue of escherichia coli. none of the chimeric pro ... | 1994 | 7830717 |
| characterization of bkdr-dna binding in the expression of the bkd operon of pseudomonas putida. | the bkd operon of pseudomonas putida consists of the structural genes encoding the components of the inducible branched-chain ketoacid dehydrogenase. bkdr, a positive regulator of the bkd operon and a homolog of lrp of escherichia coli is encoded by a structural gene adjacent to, and divergently transcribed from, the bkd operon of p. putida. bkdr was purified from e. coli containing bkdr cloned into pcytexp1, an expression vector. the molecular weight of bkdr obtained by gel filtration indicates ... | 1995 | 7836297 |
| cloning and sequencing of a b. subtilis sigmaf dependent gene from b. megaterium. | a promoter-monocistronic structural gene complex from genomic dna of bacillus megaterium has been isolated and sequenced. the activity of the promoter during sporulation was measured in b. subtilis using a fusion with the xyle gene of pseudomonas putida which codes for a catechol-2,3-dioxygenase. from the time of activation in sporulating cells and the activity in a set of defined b. subtilis sporulation mutants we conclude that the promoter requires an active sigmaf-factor of rna-polymerase. si ... | 1994 | 7842232 |
| amplification of the groesl operon in pseudomonas putida increases siderophore gene promoter activity. | pseudobactin 358 is the yellow-green fluorescent siderophore [microbial iron(iii) transport agent] produced by pseudomonas putida wcs358 under iron-limiting conditions. the genes encoding pseudobactin 358 biosynthesis are iron-regulated at the level of transcription. in this study, the molecular characterization is reported of a cosmid clone of wcs358 dna that can stimulate, in an iron-dependent manner, the activity of a wcs358 siderophore gene promoter in the heterologous pseudomonas strain a22 ... | 1994 | 7845355 |
| oxidation of 2-methoxynaphthalene by toluene, naphthalene and biphenyl dioxygenases:structure and absolute stereochemistry of metabolites. | 2-methoxynaphthalene was subjected to biooxidation by whole cells of six organisms: pseudomonas putida f39/d containing toluene dioxygenase, escherichia coli jm109(pdtg601), containing recombinant toluene dioxygenase from pp f39/d, pseudomonas sp. ncib 9816/11, containing naphthalene dioxygenase. e. coli jm109(pdtg141), containing recombinant naphthalene dioxygenase from ncib 98161/11, e. coli c534(pror/sac) containing recombinant naphthalene dioxygenase from pp g7, and beijerinckia sp. b8/36, c ... | 1994 | 7858982 |
| variability of peptidoglycan structural parameters in gram-negative bacteria. | muropeptide composition of peptidoglycan from the gram-negative bacteria aeromonas sp., acinetobacter acetoaceticus, agrobacterium tumefaciens, enterobacter cloacae, proteus morganii, pseudomonas aeruginosa, pseudomonas putida, vibrio parahaemolyticus yersinia enterocolitica and escherichia coli, was analyzed by hplc. in all instances peptidoglycan was built up from the same subunits. a wide disparity in the relative abundance of muropeptides and all structural parameters was observed. the contr ... | 1995 | 7867925 |
| overlapping substrate specificities of benzaldehyde dehydrogenase (the xylc gene product) and 2-hydroxymuconic semialdehyde dehydrogenase (the xylg gene product) encoded by tol plasmid pww0 of pseudomonas putida. | two aldehyde dehydrogenases involved in the degradation of toluene and xylenes, namely, benzaldehyde dehydrogenase and 2-hydroxymuconic semialdehyde dehydrogenase, are encoded by the xylc and xylg genes, respectively, on tol plasmid pww0 of pseudomonas putida. the nucleotide sequence of xylc was determined in this study. a protein exhibiting benzaldehyde dehydrogenase activity had been purified from cells of p. putida (pww0) (j. p. shaw and s. harayama, eur. j. biochem. 191:705-714, 1990); howev ... | 1995 | 7868591 |
| a manganese-dependent dioxygenase from arthrobacter globiformis cm-2 belongs to the major extradiol dioxygenase family. | almost all bacterial ring cleavage dioxygenases contain iron as the catalytic metal center. we report here the first available sequence for a manganese-dependent 3,4-dihydroxyphenylacetate (3,4-dhpa) 2,3-dioxygenase and its further characterization. this manganese-dependent extradiol dioxygenase from arthrobacter globiformis cm-2, unlike iron-dependent extradiol dioxygenases, is not inactivated by hydrogen peroxide. also, ferrous ions, which activate iron extradiol dioxygenases, inhibit 3,4-dhpa ... | 1995 | 7868595 |
| lipase modulator protein (liml) of pseudomonas sp. strain 109. | plasmids containing a pseudomonas sp. strain 109 extracellular lipase gene (lipl) lacking nh2-terminal sequence and a lipase modulator gene (liml) lacking the nh2-terminal hydrophobic region were constructed and expressed independently in escherichia coli by using the t7 promoter expression vector system. recombinant lipl (rlipl) was produced as inclusion bodies, whereas recombinant liml (rliml) was present as a soluble protein. during in vitro renaturation of the purified rlipl inclusion bodies ... | 1995 | 7868599 |
| 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 | |
| regulation of the hema gene during 5-aminolevulinic acid formation in pseudomonas aeruginosa. | the general tetrapyrrole precursor 5-aminolevulinic acid is formed in bacteria via two different biosynthetic pathways. members of the alpha group of the proteobacteria use 5-aminolevulinic acid synthase for the condensation of succinyl-coenzyme a and glycine, while other bacteria utilize a two-step pathway from aminoacylated trna(glu). the trna-dependent pathway, involving the enzymes glutamyl-trna reductase (encoded by hema) and glutamate-1-semialdehyde-2,1-aminomutase (encoded by heml), was d ... | 1995 | 7883699 |
| aromatic effector activation of the ntrc-like transcriptional regulator phhr limits the catabolic potential of the (methyl)phenol degradative pathway it controls. | pseudomonas putida p35x (ncib 9869) metabolizes phenol and monomethylphenols via a chromosomally encoded meta-cleavage pathway. we have recently described a 13.4-kb fragment of the chromosome that codes for the first eight genes of the catabolic pathway and a divergently transcribed positive regulator, phhr. the eight structural genes lie in an operon, the phh operon, downstream of a -24 tggc, -12 ttgc promoter sequence. promoters of this class are recognized by rna polymerase that utilizes the ... | 1995 | 7883704 |
| single amino acids changes in the signal receptor domain of xylr resulted in mutants that stimulate transcription in the absence of effectors. | the xylr protein positively controls expression from the pseudomonas putida tol plasmid sigma 54-dependent "upper" pathway operon promoter (pu) and the xyls gene promoter (ps), in response to the presence of aromatic effectors. two mutant xylr regulators able to stimulate transcription from pu and ps in the absence of effectors were isolated. these mutants exhibited single point mutations, namely asp135-->asn and pro85-->ser. both mutations are located in the amino termini domain of xylr, which ... | 1995 | 7890623 |
| identification of functional residues in a 2-hydroxymuconic semialdehyde hydrolase. a new member of the alpha/beta hydrolase-fold family of enzymes which cleaves carbon-carbon bonds. | the 2-hydroxymuconic semialdehyde hydrolase, xylf, of the pseudomonas putida tol plasmid-encoded pathway for the catabolism of toluene and xylenes, catalyzes one of the rarest types of enzyme reaction (ec 3.7.1.9), the hydrolysis of a carbon-carbon bond in its substrate, the ring-fission product of 3-alkyl-substituted catechols. in this study, amino acid sequence comparisons between xylf and other hydrolases, and analysis of the similarity between the predicted secondary structure of xylf and th ... | 1995 | 7890778 |
| mechanism of the reaction catalyzed by mandelate racemase: structure and mechanistic properties of the k166r mutant. | on the basis of the available high-resolution structures of mandelate racemase (mr) from pseudomonas putida [landro, j. a., gerlt, j. a., kozarich, j. w., koo, c. w., shah, v. j., kenyon, g. l., neidhart, d. j., fujita, j., & petsko, g. a. (1994) biochemistry 33, 635-643], lys 166 and his 297 are positioned appropriately to participate in catalysis as acid/base catalysts that either abstract the alpha-proton from the enantiomers of mandelate to form an enolic intermediate or protonate the enolic ... | 1995 | 7893690 |
| aspartate transcarbamoylase genes of pseudomonas putida: requirement for an inactive dihydroorotase for assembly into the dodecameric holoenzyme. | the nucleotide sequences of the genes encoding the enzyme aspartate transcarbamoylase (atcase) from pseudomonas putida have been determined. our results confirm that the p. putida atcase is a dodecameric protein composed of two types of polypeptide chains translated coordinately from overlapping genes. the p. putida atcase does not possess dissociable regulatory and catalytic functions but instead apparently contains the regulatory nucleotide binding site within a unique n-terminal extension of ... | 1995 | 7896697 |
| a carbon starvation survival gene of pseudomonas putida is regulated by sigma 54. | by using mini-tn5 transposon mutagenesis, two mutants of pseudomonas putida atcc 12633 were isolated which showed a marked increase in their sensitivity to carbon starvation; these mutants are presumably affected in the pex type of proteins that p. putida induces upon carbon starvation (m. givskov, l. eberl, and s. molin, j. bacteriol. 176:4816-4824, 1994). the affected genes in our mutants were induced about threefold upon carbon starvation. the promoter region of the starvation gene in the mut ... | 1995 | 7896711 |
| cloning, expression and mutational analysis of the urocanase gene (hutu) from pseudomonas putida. | the histidine-utilizing hutu gene was isolated from a lambda-embl3 phage of a genomic library from pseudomonas putida nicii and subcloned into the expression vector pt7-7. escherichia coli bl21 cells were transformed with the recombinant plasmid and produced a catalytically active protein, amounting to approximately 30% of the total protein in the crude cell-free extract. the addition of nad+ to the growth medium ensured the full occupation of active sites by the cofactor. this requires a mechan ... | 1993 | 7901006 |
| 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 |
| transcriptional control of the pseudomonas putida tol plasmid catabolic pathways. | tol plasmid pww0 of pseudomonas putida contains two operons that specify a pathway for the degradation of aromatic hydrocarbons. the upper pathway operon encodes the enzymes for the oxidation of toluene/xylenes to benzoate/toluates, and the metacleavage pathway operon encodes the enzymes for the further oxidation of these compounds to krebs cycle intermediates. their expression is controlled by the gene products of two divergently transcribed regulatory genes, xyir and xyis. the xyir protein, wh ... | 1993 | 7934920 |
| 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 |
| identification and characterization of a siderophore regulatory gene (pfra) of pseudomonas putida wcs358: homology to the alginate regulatory gene algq of pseudomonas aeruginosa. | genes encoding biosynthesis of pseudobactin 358 (a microbial iron transport agent) and its cognate outer membrane receptor protein, pupa, are transcribed only under iron limitation in plant growth-promoting pseudomonas putida wcs358. two cosmid clones were identified from a gene bank of wcs358 dna which could independently and in an iron-dependent manner activate transcription from a wcs358 siderophore gene promoter in heterologous pseudomonas strain a225. the functional region of one of the clo ... | 1993 | 7968519 |
| 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 |
| in vitro and in situ survivals of bacterial populations added to fresh water environments. | the fate of aeromonas hydrophila, alcaligenes denitrificans, vibrio cholerae non-01, pseudomonas putida and four different isolates of escherichia coli in fresh river water were assessed by using different microcosms (i.e., membrane diffusion chamber and erlenmeyer flask). when water samples were incubated at 16 +/- 1 degrees c, the differences in extent of survival among test bacteria were in general not significant. if the incubation temperature was raised to 29 +/- 1 degrees c, in the in situ ... | 1993 | 7982366 |
| 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 |