Publications
| Title | Abstract | Year Filter | PMID(sorted ascending) Filter |
|---|
| construction of phoe-caa, a novel pcr- and immunologically detectable marker gene for pseudomonas putida. | in this paper we describe the construction and use in pseudomonas putida wcs358 of phoe-caa, a novel hybrid marker gene, which allows monitoring both at the protein level by immunological methods and at the dna level by pcr. the marker is based on the escherichia coli outer membrane protein gene phoe and 75 bp of e. coli caa, which encode a nonbacteriocinic fragment of colicin a. this fragment contains an epitope which is recognized by monoclonal antibody (mab) 1c11. as the epitope is contained ... | 1994 | 7993086 |
| metabolism of chlorofluorocarbons and polybrominated compounds by pseudomonas putida g786(phg-2) via an engineered metabolic pathway. | the recombinant bacterium pseudomonas putida g786(phg-2) metabolizes pentachloroethane to glyoxylate and carbon dioxide, using cytochrome p-450cam and toluene dioxygenase to catalyze consecutive reductive and oxidative dehalogenation reactions (l.p. wackett, m.j. sadowsky, l.n. newman, h.-g. hur, and s. li, nature [london] 368:627-629, 1994). the present study investigated metabolism of brominated and chlorofluorocarbon compounds by the recombinant strain. under anaerobic conditions, p. putida g ... | 1994 | 7993096 |
| loss of the tol meta-cleavage pathway functions of pseudomonas putida strain paw1 (pww0) during growth on toluene. | a derivative of pseudomonas putida strain paw1 bearing the tol plasmid pww0 was isolated from a culture which has grown unlimited on toluene. in contrast to the parent strain paw1, the derivative, strain cg220, is unable to grow with xylenes and toluates, while toluene and benzoate served as substrates. strain cg220 had a remarkable growth advantage against the wild type when grown with toluene. biochemical analysis showed that in strain cg220 toluene was metabolised through the tol plasmid uppe ... | 1994 | 7996396 |
| permeabilization of mycolic-acid-containing actinomycetes for in situ hybridization with fluorescently labelled oligonucleotide probes. | the application of whole-cell hybridization using labelled oligonucleotide probes in microbial systematics and ecology is limited by difficulties in permeabilizing many gram-positive organisms. in this investigation paraformaldehyde treatment, acid methanolysis and acid hydrolysis were evaluated as a means of permeabilizing mycolic-acid-containing actinomycetes prior to hybridization with a fluorescently labelled oligonucleotide probe designed to bind to a conserved sequence of bacterial 16s rrn ... | 1994 | 8000549 |
| enzymatic asymmetric synthesis of alpha-methyl arylalkylamines and alpha-methyl arylalkylalcohols by arylalkyl acylamidases. | with the novel microbial enzyme, 'arylalkyl acylamidase', optically active alpha-methyl arylalkylamines and alpha-methyl arylalkylalcohols have been obtained through enantioselective hydrolysis of their racemic amides and esters. (s)-enantiomers of 1-methylbenzylamine, 1-methyl-3-phenylpropylamine and 1-methyl-3-phenylpropanol of high optical purity (> 94% e.e.) were synthesized with the cells of nocardia erythropolis iam 1440 or cellulomonas fimi aku 671. (r)-enantiomer of 1-methyl-3-phenylprop ... | 1994 | 8000864 |
| combination effect of meropenem with aminoglycosides and teicoplanin on pseudomonas and enterococci. | the in vitro activity of meropenem, a new carbapenem, and the combination effect with netilmicin, tobramycin, gentamicin, and teicoplanin against pseudomonas spp. and enterococci was studied. meropenem showed very good in vitro activity against pseudomonas aeruginosa (mic90 2 mg/l) and good to moderate activity against pseudomonas putida (mic90 4 mg/l) and enterococcus faecalis (mic90 8 mg/l). aminoglycosides were highly active against p. putida (mic90 0.5 mg/l), but showed only moderate activit ... | 1994 | 8002095 |
| oxidative release of nitrite from 2-nitrotoluene by a three-component enzyme system from pseudomonas sp. strain js42. | pseudomonas sp. strain js42 utilizes 2-nitrotoluene (2nt) as the sole source of carbon and energy for growth. intact cells catalyze the oxidation of 2nt to 3-methylcatechol and nitrite in a reaction that requires molecular oxygen. cell extracts oxidized 2nt to 3-methylcatechol and nitrite in the presence of nad(p)h and ferrous iron. ion-exchange chromatography yielded three protein fractions (a, b, and c) which were all required for the oxidation of 2nt to 3-methylcatechol and nitrite. component ... | 1994 | 8002568 |
| aerobic catabolism of phenylacetic acid in pseudomonas putida u: biochemical characterization of a specific phenylacetic acid transport system and formal demonstration that phenylacetyl-coenzyme a is a catabolic intermediate. | the phenylacetic acid transport system (pats) of pseudomonas putida u was studied after this bacterium was cultured in a chemically defined medium containing phenylacetic acid (pa) as the sole carbon source. kinetic measurement was carried out, in vivo, at 30 degrees c in 50 mm phosphate buffer (ph 7.0). under these conditions, the uptake rate was linear for at least 3 min and the value of km was 13 microm. the pats is an active transport system that is strongly inhibited by 2,4-dinitrophenol, 4 ... | 1994 | 8002592 |
| organization and evolution of naphthalene catabolic pathways: sequence of the dna encoding 2-hydroxychromene-2-carboxylate isomerase and trans-o-hydroxybenzylidenepyruvate hydratase-aldolase from the nah7 plasmid. | the sequence of a 2,437-bp dna segment from the naphthalene upper catabolic pathway operon of plasmid nah7 was determined. this segment contains three large open reading frames designated nahq', nahe, and nahd. the first of these is the 3' end of an open reading frame that has no known function, the second (993 bp) encodes trans-o-hydroxybenzylidenepyruvate hydratase-aldolase (deduced molecular weight, 36,640), and the third (609 bp) encodes 2-hydroxychromene-2-carboxylate isomerase (deduced mol ... | 1994 | 8002605 |
| codon usage patterns suggest independent evolution of two catabolic operons on toluene-degradative plasmid tol pww0 of pseudomonas putida. | tol plasmid pww0 of pseudomonas putida encodes a set of enzymes responsible for the degradation of toluene. the structural genes for these catobolic enzymes are clustered into two operons--namely, the xy/cmab and xy/xyzltegfjqkih operons. we examined the codon usage patterns of these catabolic genes by measuring the codon-usage distances between pairs of these catabolic genes. the codon-usage distance, d, between gene 1 and gene 2 was defined as d = [sigma(pj-qj)2]1/2, are the frequencies of the ... | 1994 | 8007001 |
| reaction engineering aspects of conjugation in biodegradation processes. | conjugation between two pseudomonas putida species has been studied. special emphasis has been given to the design of tools for better defined and reproducible experimental conditions. this is an essential prerequisite for any kinetic analysis of the phenomenon. the experimental approach suggested in the paper is a three-stage continuous fermentation, where donor and recipient strains can be cultivated at predefined steady-state growth rates, and conjugation takes place at steady-state condition ... | 1994 | 8010691 |
| 1-amino-2-imidazol-4'-ylethylphosphonic acid is a potent reversible inhibitor of pseudomonas putida histidine ammonia-lyase. | a phosphonic acid analogue of l-histidine, 1-amino-2-imidazol-4'-ylethylphosphonic acid (hisp), was identified as a reversible competitive inhibitor of histidine ammonia-lyase (histidase). the affinity of histidase for hisp was ph dependent, with ki values of 0.28 microm and 10.4 microm compared to substrate km values of 1 and 5 mm at ph 7 and 9, respectively. hisp did not appear to be a substrate for histidase. a twenty-fold molar excess of hisp over enzyme completely protected the active site ... | 1994 | 8012285 |
| transposon-mediated mobilization of chromosomally located catabolic operons of the cam plasmid by tol plasmid transposon tn4652 and cam plasmid transposon tn3614. | the cam (camphor degradation) plasmid is integrated into the chromosome of pseudomonas putida paw-line strains and is not self-transferable as a plasmid via conjugation. our results show that the mobilization of chromosomally located cam and the integration of cam-operons into the chromosome of the new cam+ transconjugants is a reca-independent process mediated by transposons tn4652 (17 kbp) and tn3614 (7.2 kbp). transposon tn3614 is apparently identical to the left-hand and the right-hand seque ... | 1994 | 8012608 |
| siderophore receptor pupa as a marker to monitor wild-type pseudomonas putida wcs358 in natural environments. | for application of genetically engineered fluorescent pseudomonas spp., specific markers are required for monitoring of wild-type pseudomonas strains and their genetically modified derivatives in natural environments. in this study, the specific siderophore receptor pupa of plant growth-promoting pseudomonas putida wcs358 was used as a marker to monitor wild-type strain wcs358. after introduction into natural soil and rhizosphere environments, strain wcs358 could be recovered efficiently on a me ... | 1994 | 8017914 |
| quantitative cell lysis of indigenous microorganisms and rapid extraction of microbial dna from sediment. | this study reports improvements in two of the key steps, lysis of indigenous cells and dna purification, required for achieving a rapid nonselective protocol for extracting nucleic acids directly from sodium dodecyl sulfate (sds)-treated sediment rich in organic matter. incorporation of bead-mill homogenization into the dna extraction procedure doubled the densitometrically determined dna yield (11.8 micrograms of dna.g [dry weight] of sediment-1) relative to incorporation of three cycles of fre ... | 1994 | 8017936 |
| functional and structural relationship of various extradiol aromatic ring-cleavage dioxygenases of pseudomonas origin. | the extradiol ring-cleavage dioxygenases derived from seven different pseudomonas strains were expressed in escherichia coli and the substrate specificities were investigated for a variety of catecholic compounds. the substrate range of four 2,3-dihydroxybiphenyl dioxygenases from biphenyl-utilizing bacteria, 3-methylcatechol dioxygenase from toluene utilizing pseudomonas putida f1, 1,2-dihydroxynaphthalene dioxygenase from a nah7 plasmid, and catechol 2,3-dioxygenase from a tol plasmid pww0 wer ... | 1994 | 8020752 |
| biotransformation of benzothiophene by isopropylbenzene-degrading bacteria. | isopropylbenzene-degrading bacteria, including pseudomonas putida re204, transform benzothiophene to a mixture of compounds. induced strain re204 and a number of its tn5 mutant derivatives were used to accumulate these compounds and their precursors from benzothiophene. these metabolites were subsequently identified by 1h and 13c nuclear magnetic resonance spectroscopy and gas chromatography-mass spectrometry. when strain re204 was incubated with benzothiophene, it produced a bright yellow compo ... | 1994 | 8021182 |
| inability of muconate cycloisomerases to cause dehalogenation during conversion of 2-chloro-cis,cis-muconate. | the conversion of 2-chloro-cis,cis-muconate by muconate cycloisomerase from pseudomonas putida prs2000 yielded two products which by nuclear magnetic resonance spectroscopy were identified as 2-chloro- and 5-chloromuconolactone. high-pressure liquid chromatography analyses showed the same compounds to be formed also by muconate cycloisomerases from acinetobacter calcoaceticus adp1 and pseudomonas sp. strain b13. during 2-chloro-cis,cis-muconate turnover by the enzyme from p. putida, 2-chloromuco ... | 1994 | 8021223 |
| ser-143 is an essential active site residue in histidine ammonia-lyase of pseudomonas putida. | site directed mutagenesis was used to investigate the role of ser-143 in enzyme activity and as a point for attack by cyanide or l-cysteine, two irreversible inhibitors of histidine ammonia-lyase (histidase). two mutant proteins, a s143a substitution and an a142s-s143a transposition, were made. both mutant histidases completely lost all enzymatic activity. western blots with anti-histidase antibodies revealed that the mutant proteins were being expressed at a level equal to that of the wild-type ... | 1994 | 8024588 |
| cloning, heterologous expression, and sequencing of a novel proline iminopeptidase gene, pepi, from lactobacillus delbrueckii subsp. lactis dsm 7290. | the gene for proline iminopeptidase from lactobacillus delbrueckii subsp. lactis dsm 7290 coding for an enzyme that hydrolyses the synthetic substrate l-prolyl-beta-naphthylamide (pro-beta na) was cloned in escherichia coli. an enzymic plate assay was used to screen for positive clones. the gene, designated pepi, was subcloned into vector puc18 and sequenced. the nucleotide sequence revealed an 882 bp open reading frame encoding 294 amino acids, coding for an enzyme with a calculated molecular m ... | 1994 | 8025678 |
| role for the outer membrane ferric siderophore receptor pupb in signal transduction across the bacterial cell envelope. | the outer membrane protein pupb of pseudomonas putida wcs358 facilitates transport of iron complexed to the siderophores pseudobactin bn8 and pseudobactin bn7 into the cell. its synthesis is induced by the presence of these specific siderophores under iron limitation. the signal transduction pathway regulating siderophore-dependent expression of pupb was shown to consist of two regulatory proteins, pupi and pupr, and the pupb receptor itself. mutational analysis of the regulatory genes suggested ... | 1994 | 8026465 |
| purification of a sixth ferredoxin from rhodobacter capsulatus. primary structure and biochemical properties. | a new ferredoxin has been purified from the photosynthetic bacterium rhodobacter capsulatus. it is the sixth ferredoxin to be isolated from this bacterium and it was called fdvi. its primary structure was established based on amino acid sequence analysis of the protein and of peptides derived from it. it is composed of 106 residues including five cysteines. the calculated mass of the polypeptide is 11,402.6 da which matches the experimental value determined by electrospray mass spectrometry. ami ... | 1994 | 8026503 |
| analysis of fluorescent pseudomonads based on 23s ribosomal dna sequences. | the regions from positions 1391 to 1545 and 1620 to 1865 (escherichia coli numbers) of the 23s ribosomal dna sequences have been analyzed for a number of pseudomonas fluorescens and p. putida isolates. variability was observed only in three smaller regions from positions 1484 to 1508, 1531 to 1542, and 1714 to 1748, corresponding to helices 58, 59, and 63, respectively, where up to 53% dissimilarity was found. sequence analysis did not allow clear distinctions among p. fluorescens biovars, p. ch ... | 1994 | 8031104 |
| nucleotide sequence of the gene encoding a repressor for the cytochrome p-450cam hydroxylase operon on the pseudomonas putida cam plasmid. | the camr gene of pseudomonas putida encodes a repressor which regulates expression of the cytochrome p-450cam hydroxylase operon (camdcab). we determined the nucleotide sequence of 1134 continuous base pairs, including the camr gene. when comparing the amino acid sequence deduced from the open reading frame of the gene sequence with that of amino-terminal five residues of the cam repressor, purified from pseudomonas putida, we found that the camr gene encodes a protein of 186 amino acids, with a ... | 1994 | 8031906 |
| purification and characterization of morphinone reductase from pseudomonas putida m10. | the nadh-dependent morphinone reductase from pseudomonas putida m10 catalyses the reduction of morphinone and codeinone to hydromorphone and hydrocodone respectively. morphinone reductase was purified from crude cell extracts to apparent homogeneity in a single affinity-chromatography step using mimetic yellow 2. the purified enzyme was a dimeric flavoprotein with two identical subunits of m(r) 41,100, binding non-covalently one molecule of fmn per subunit. the n-terminal sequence was pdtsfsnpgl ... | 1994 | 8037698 |
| kinetics and thermodynamics of co binding to cytochrome p450nor. | the co-binding reaction of cytochrome p450nor isolated from denitrifying fungus, fusarium oxysporum, has been studied by using a flash photolysis method in the millisecond time domain. we obtained the co on- and off-rate constants in the bimolecular reaction, and determined the activation free energy, enthalpy, and entropy from the temperature dependence of these rate constants. to discuss the structural characteristics of p450nor, these parameters were compared with those of other cytochrome p4 ... | 1994 | 8038156 |
| evidence for the involvement of multiple pathways in the biodegradation of 1- and 2-methylnaphthalene by pseudomonas putida csv86. | pseudomonas putida csv86, a soil bacterium, grows on 1- and 2-methylnaphthalene as the sole source of carbon and energy. in order to deduce the pathways for the biodegradation of 1- and 2-methylnaphthalene, metabolites were isolated from the spent medium and purified by thin layer chromatography. emphasis has been placed on the structural characterisation of isolated intermediates by gc-ms, demonstration of enzyme activities in the cell free extracts and measurement of oxygen uptake by whole cel ... | 1994 | 8042906 |
| significant contribution of arginine-112 and its positive charge of pseudomonas putida cytochrome p-450cam in the electron transport from putidaredoxin. | cytochrome p-450cam of pseudomonas putida is a prototype of various eukaryotic cytochrome p-450 molecules. arg-112 located on the surface of this protein is highly conserved among various other cytochromes p-450. in this study, we constructed mutant genes for p-450cam in which arg-112 was replaced by gln or glu, expressed them in escherichia coli and purified the mutant proteins. their enzymic activities were analyzed in the reconstituted system to determine the function of arg-112. kd values fo ... | 1994 | 8043608 |
| [mossbauer data on the effect of nad.h on the state of iron in bacteria]. | the state of fe of bacterial cultures of different systematic positions (bacillus megaterium, bacillus polymyxa, pseudomonas putida, pseudomonas fluorescens, alcaligenes faecalis, arthrobacter siderocapsulatus) grown on the medium containing fe(iii) citrate (up to 100 mg/l) with additional or without nad.h was studied. the samples were in damp air-dry, second moistened, dried at 383 k states. spectra have been obtained at 290 k and 100-200 k. the studied microorganisms have two types of atoms of ... | 1994 | 8043633 |
| [controlled-expression vector for pseudomonas bacteria involving regulatory elements of trpiba genes of pseudomonas putida]. | a bireplicon controlled-expression vector pps10 was developed based, on trpiba genes of pseudomonas putida. it is a low-copy-number vector in pseudomonas bacteria, and a high-copy-number vector in escherichia coli. the vector is 10.4 kilobase pairs (kb), determines resistance to kanamycin, carries a replicon of cryptic pseudomonas pmk1 plasmid; a pbr322 replicon; the par locus of pmt2 plasmid; and the trpi gene of p. putida, which encodes the activator protein and the promoter pba of trpba genes ... | 1994 | 8045378 |
| chromosomal gene capture mediated by the pseudomonas putida tol catabolic plasmid. | the pseudomonas putida tol plasmid pww0 is able to mediate chromosomal mobilization in the canonical unidirectional way, i.e., from donor to recipient cells, and bidirectionally, i.e., donor-->recipient-->donor (retrotransfer). transconjugants are recipient cells that have received dna from donor cells, whereas retrotransconjugants are donor bacteria that have received dna from a recipient. the tol plasmid pww0 is able to directly mobilize and retromobilize a kanamycin resistance marker integrat ... | 1994 | 8045894 |
| responses to nutrient starvation in pseudomonas putida kt2442: two-dimensional electrophoretic analysis of starvation- and stress-induced proteins. | the responses of pseudomonas putida kt2442 to various forms of nutrient starvation and stress conditions were examined by two-dimensional polyacrylamide electrophoresis. carbon deprivation resulted in a temporal expression of two classes of starvation-induced proteins: one class was transiently expressed during the initial phase of starvation, and the second class was expressed throughout the entire starvation period. proteins of the second class could be further subdivided into proteins induced ... | 1994 | 8050994 |
| cross-regulation by xylr and dmpr activators of pseudomonas putida suggests that transcriptional control of biodegradative operons evolves independently of catabolic genes. | the pu promoter of the toluene degradation plasmid pww0 of pseudomonas putida drives expression of an operon involved in the sequential oxidation of toluene and m- and p-xylenes to benzoate and toluates, respectively. similarly, the po promoter of plasmid pvi150 controls expression of an operon of pseudomonas sp. strain cf600 which is required for the complete catabolism of phenol and cresols. these promoters, which both belong to the sigma 54-dependent class, are regulated by their cognate acti ... | 1994 | 8051017 |
| tartrate dehydrogenase, a new member of the family of metal-dependent decarboxylating r-hydroxyacid dehydrogenases. | the gene encoding tartrate dehydrogenase has been cloned from pseudomonas putida and sequenced. the gene is 1098 nucleotides long and encodes a protein 365 amino acids in length with a calculated m(r) of 40,636. the gene and the protein encoded by it show strong homology to prokaryotic isopropylmalate dehydrogenases and, to a lesser extent, isocitrate dehydrogenase. thus, tartrate dehydrogenase is the third member to be identified of the family of metal-dependent decarboxylating r-hydroxyacid de ... | 1994 | 8053675 |
| beta-ureidopropionase with n-carbamoyl-alpha-l-amino acid amidohydrolase activity from an aerobic bacterium, pseudomonas putida ifo 12996. | beta-ureidopropionase of aerobic bacterial origin was purified to homogeneity from pseudomonas putida ifo 12996. the enzyme shows a broad substrate specificity. in addition to beta-ureidopropionate (km 3.74 mm, vmax 4.12 u/mg), gamma-ureido-n-butyrate (km 11.6 mm, vmax 19.4 u/mg), and several n-carbamoyl-alpha-amino acids, such as n-carbamoylglycine (km 0.68 mm, vmax 9.14 x 10(-2) u/mg), n-carbamoyl-l-alanine (km 1.56 mm, vmax 1.00 u/mg), n-carbamoyl-l-serine (km 75.1 mm, vmax 3.78 u/mg), and n- ... | 1994 | 8055933 |
| sequence and expression of the todgih genes involved in the last three steps of toluene degradation by pseudomonas putida f1. | the todfc1c2bade gene cluster in pseudomonas putida f1 encodes enzymes for the first four steps of toluene degradation, leading to the formation of 2-hydroxypenta-2,4-dienoate (hpd). here, we report the nucleotide (nt) sequence and expression of the remaining three genes of the tod pathway, downstream from tode and arranged in the order, todgih. the deduced amino acid (aa) sequences of todg [hpd hydratase (268 aa)], todh [4-hydroxy-2-oxovalerate (ho) aldolase (352 aa)] and todi [acylating aldehy ... | 1994 | 8063106 |
| substrate, substrate analogue, and inhibitor interactions with the ferrous active site of catechol 2,3-dioxygenase monitored through xas studies. | the interactions of catechol (substrate), 2-hydroxy-pyridine-n-oxide (substrate analogue), and 2-bromophenol (inhibitor) with the extradiol cleaving catechol-2,3-dioxygenase from pseudomonas putida mt-2 have been monitored through x-ray absorption spectroscopy (xas). the analysis of the data provides details about the mode of coordination of the substrate and of the inhibitors to the active site of the enzyme. | 1994 | 8070565 |
| rhizobia catabolize nod gene-inducing flavonoids via c-ring fission mechanisms. | gas chromatographic and mass spectrometric analyses of derivatized culture medium extracts were used to identify the products of flavonoid metabolism by rhizobia. a number of rhizobium species and biovars degraded their nod gene-inducing flavonoids by mechanisms which originated in a cleavage of the c-ring of the molecule and which yielded conserved a- and b-ring products among the metabolites. in contrast, pseudomonas putida degraded quercetin via an initial fission in its a-ring, and agrobacte ... | 1994 | 8071218 |
| purification of the lysr family regulator, clcr, and its interaction with the pseudomonas putida clcabd chlorocatechol operon promoter. | previous studies have shown that the clcabd operon is under the transcriptional control of the lysr-type activator clcr. in this study, the conditions leading to its aggregation were avoided and clcr was purified and confirmed by amino-terminal sequencing. gel filtration indicated that clcr exists as a dimer in solution. the dnase i footprint of clcr was determined. the binding properties of clcr and the catechol operon regulator, catr, were compared. | 1994 | 8071232 |
| cross talk between catabolic pathways in pseudomonas putida: xyls-dependent and -independent activation of the tol meta operon requires the same cis-acting sequences within the pm promoter. | the pm promoter of the meta cleavage operon in the tol (toluene degradation) plasmid pww0 of pseudomonas putida becomes activated by the plasmid-encoded xyls regulator in the presence of benzoate and certain substituted analogs such as 3-methylbenzoate. in the absence of xyls, pm was still responsive to unsubstituted benzoate but with induction kinetics and a range of transcriptional activity which differed substantially from those for the xyls-mediated activation. xyls-independent induction by ... | 1994 | 8071244 |
| trichloroethylene and chloroform degradation by a recombinant pseudomonad expressing soluble methane monooxygenase from methylosinus trichosporium ob3b. | soluble methane monooxygenase (smmo) from methylosinus trichosporium ob3b can degrade many halogenated aliphatic compounds that are found in contaminated soil and groundwater. this enzyme oxidizes the most frequently detected pollutant, trichloroethylene (tce), at least 50 times faster than other enzymes. however, slow growth of the strain, strong competition between tce and methane for smmo, and repression of the smmo locus by low concentrations of copper ions limit the use of this bacterium. t ... | 1994 | 8074526 |
| x-ray absorption studies on catechol 2,3-dioxygenase from pseudomonas putida mt2. | x-ray absorption spectroscopy has been utilized to investigate the structure of the active site of iron(ii) catechol 2,3-dioxygenase from pseudomonas putida mt2 both in the native and the 2-chlorophenol inhibited forms. xanes (x-ray absorption near edge structure) and exafs (extended x-ray absorption fine structure) results allow us to discuss the coordination number and geometry of the ferrous ion in the native enzyme. the metal geometry is not significantly affected by the binding of the inhib ... | 1994 | 8075079 |
| purification and characterization of a novel metal-containing nonheme bromoperoxidase from pseudomonas putida. | a novel bromoperoxidase was purified to homogeneity from the bacterium pseudomonas putida if-3 strain, which produces the antibiotic pyrrolnitrin. the enzyme had a molecular mass of 68,000 and was composed of two identical subunits (33,000). it was specific for i- and br- and inactive toward cl- and f- in the monochlorodimedone assay system. the optimum ph of the enzyme was around 4.2 and it rapidly lost its activity below 3.5, but it was stable over of range ph of 4 to 11. the purified enzyme w ... | 1994 | 8075154 |
| identification of variability of ribosomal dna spacer from pseudomonas soil isolates. | the polymerase chain reaction was used to amplify the spacer region located between the 16s and 23s ribosomal rna genes of strains of pseudomonas fluorescens and pseudomonas putida isolated from peat bog, canola field, or arctic plants. some of spacer region of four of the p. fluorescens strains examined, strains 64-3, 63-28, qp5, and r17-fp2, was about 515 base pairs (bp) in length, and contained the genes for trna(ile) and trna(ala). the dna sequences of two strains from canola, 64-3 and 63-28 ... | 1994 | 8076249 |
| co-regulation by bent dna. functional substitutions of the integration host factor site at sigma 54-dependent promoter pu of the upper-tol operon by intrinsically curved sequences. | the role of integration host factor (ihf) in the regulation of the sigma 54-dependent promoter pu of the tol plasmid of pseudomonas putida has been examined. we have selected in vivo insertions of intrinsically curved dna that restore the responsiveness of an ihf-binding site deletion variant of pu to the cognate activator of the system, xylr. we found five pu derivatives which had inserted a core sequence with 6 phased [a]6 tracts, flanked by different lengths of dna at the location of the form ... | 1994 | 8077217 |
| molecular characterization of 4-hydroxyphenylacetate 3-hydroxylase of escherichia coli. a two-protein component enzyme. | the nucleotide sequences of the hpab and hpac genes encoding the 4-hydroxyphenylacetate 3-hydroxylase from escherichia coli w atcc 11105 have been determined. these genes appear to be part of an operon and encode two proteins of 58,781 and 18,679 da, respectively, that are required for hydroxylase activity. this aromatic hydroxylase is nadh-dependent and uses fad as the redox chromophore. the largest component (hpab) has been purified by affinity chromatography in cibacron blue. e. coli cells th ... | 1994 | 8077235 |
| the vacuolar compartment is required for sulfur amino acid homeostasis in saccharomyces cerevisiae. | in order to isolate new mutations impairing transcriptional regulation of sulfur metabolism in saccharomyces cerevisiae, we used a potent genetic screen based on a gene fusion expressing xyle (from pseudomonas putida) under the control of the promoter region of met25. this selection yielded strains mutated in various different genes. we describe in this paper the properties of one of them, met27. mutation or disruption of met27 leads to a methionine requirement and affects s-adenosylmethionine ( ... | 1994 | 8078479 |
| characterization of the pcar regulatory gene from pseudomonas putida, which is required for the complete degradation of p-hydroxybenzoate. | the pca branch of the beta-ketoadipate pathway in pseudomonas putida is responsible for the complete degradation of p-hydroxybenzoate through ortho cleavage of the initial pathway metabolite, protocatechuate. the pcar regulatory locus has been found to be required for both induction of all of the genes within the pca regulon (pcabdc, pcaij, and pcaf) and the chemotactic response of the bacteria to aromatic compounds. insertional inactivation mutagenesis, using tn5 and mini-tn5 transposons, was u ... | 1994 | 8083169 |
| cometabolic oxidation of polychlorinated biphenyls in soil with a surfactant-based field application vector. | polychlorinated biphenyl (pcb)-degradative genes, under the control of a constitutive promoter, were cloned into a broad-host-range plasmid and a transposon. these constructs were inserted into a surfactant-utilizing strain, pseudomonas putida ipl5, to create a field application vector (fav) in which a surfactant-degrading organism cometabolizes pcb. by utilizing a surfactant not readily available to indigenous populations and a constitutive promoter, selective growth and pcb-degradative gene ex ... | 1994 | 8085825 |
| competition in chemostat culture between pseudomonas strains that use different pathways for the degradation of toluene. | pseudomonas putida mt-2, p. cepacia g4, p. mendocina kr1, and p. putida f1 degrade toluene through different pathways. in this study, we compared the competition behaviors of these strains in chemostat culture at a low growth rate (d = 0.05 h-1), with toluene as the sole source of carbon and energy. either toluene or oxygen was growth limiting. under toluene-limiting conditions, p. mendocina kr1, in which initial attack is by monooxygenation of the aromatic nucleus at the para position, outcompe ... | 1994 | 8085826 |
| synthesis of trans unsaturated fatty acids in pseudomonas putida p8 by direct isomerization of the double bond of lipids. | the phospholipids of pseudomonas putida p8 contain monounsaturated fatty acids in the cis and trans configuration. cells of this phenol-degrading bacterium change the proportions of these isomers in response to the addition or elimination of a membrane active compound such as 4-chlorophenol. this study undoubtedly reveals that the cis unsaturated fatty acids are directly converted into trans isomers without involvement of de novo synthesis of fatty acids. oleic acid, which cannot be synthesized ... | 1994 | 8085914 |
| the induction and repression of benzene and catechol oxidizing capacity of pseudomonas putida ml2 studied in perturbed chemostat culture. | the oxidation of catechol, an intermediate in benzene catabolism, was studied using transient variations in dissolved oxygen tension (dot) when a succinate limited steady state culture of pseudomonas putida ml2 was perturbed with a pulse of another substrate. a model was developed and tested for the effect of fluctuations in oxidizing enzyme activity on dot. it was found that the rate of induction of catechol oxidizing enzymes was independent of dilution rate up to a relative growth rate (mu/mum ... | 1994 | 8085917 |
| 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 |
| crystallization of catechol-1,2 dioxygenase from pseudomonas arvilla c-1. | the metalloenzyme catechol 1,2-dioxygenase from pseudomonas arvilla c-1 consists of three isozymes formed by combinations of two non-identical subunits; alpha alpha, alpha beta and beta beta; with molecular masses of 59,000, 63,000 and 67,000 da, respectively. the alpha alpha isozyme crystallizes in the orthorhombic space group c222(1) with unit cell dimensions a = 62.7 a, b = 71.5 a, c = 187.1 a. the rectangular plates diffract to 2.6 a resolution. this is the first dioxygenase to be crystalliz ... | 1994 | 8107120 |
| fur regulon in gram-negative bacteria. identification and characterization of new iron-regulated escherichia coli genes by a fur titration assay. | a highly sensitive genetic screen for the detection of cloned genes coding for iron-regulated and iron-storage/binding proteins was developed. the fur titration assay (furta) enabled identification of cloned iron-regulated genes from different gram-positive and gram-negative bacteria such as: bacillus subtilis, escherichia coli, pantoea agglomerans, pseudomonas putida, salmonella typhimurium, serratia marcescens and yersinia enterocolitica. an ordered e. coli cosmid library was screened for eith ... | 1994 | 8107138 |
| cis,cis-muconate lactonizing enzyme from trichosporon cutaneum: evidence for a novel class of cycloisomerases in eucaryotes. | the absolute stereochemical courses of cis,cis-muconate lactonizing enzyme (mle;ec 5.5.1.1) from trichosporon cutaneum (tcmle) and chloromuconate cycloisomerase (mle ii; ec 5.5.1.7) from pseudomonas sp b13 have been determined from 1h nmr measurements. both cycloisomerases convert cis,cis-muconate to (4s)-muconolactone by a syn lactonization, the absolute stereochemical outcome of which is identical to that observed with mle from pseudomonas putida. the regiochemical courses of cyclization of 3- ... | 1994 | 8110801 |
| mineralization of p-methyl-14c-benzoate in soils by pseudomonas putida (pww0). | pseudomonas putida bearing the archetypal tol plasmid pww0 metabolizes p-methylbenzoate through a meta-cleavage pathway. in complex environments such as soils that are relatively rich in organic matter, we observed metabolism of p-methyl-14c-benzoate, which could be monitored as 14co2 evolution from the labelled alkylaromatic. linear 14co2 evolution in soils took place for at least a month, although efficient mineralization of the alkylaromatic required appropriate mass transfer to allow the bac ... | 1993 | 8111534 |
| cloning and nucleotide sequence of the pseudomonas aeruginosa glucose-selective oprb porin gene and distribution of oprb within the family pseudomonadaceae. | oprb is a glucose-selective porin known to be produced by pseudomonas aeruginosa and pseudomonas putida. we have cloned and sequenced the oprb gene of p. aeruginosa and obtained expression of oprb in escherichia coli. the mature protein consists of 423 amino acid residues with a deduced molecular mass of 47597 da. several clusters of amino acid residues, potentially involved in the structure or function of the protein, were identified. an area of regional homology with e. coli lamb was also iden ... | 1994 | 8125108 |
| controlled-expression shuttle vector for pseudomonads based on the trpiba genes of pseudomonas putida. | a cloning vector pps7 (8.5 kb) for pseudomonas was constructed from pbr322 and the pseudomonas cryptic low-copy-number pmk1 plasmid. the vector confers resistance to kanamycin (km) and tetracycline (tc), contains the par locus of pseudomonas plasmid pmt2 and a mob site. the new vector was used for construction of controlled-expression vector pps10 (10.4 kb) based on the trpiba genes of pseudomonas putida. this kmr vector contains the trpi gene, encoding activator protein and promoter of trpba ge ... | 1994 | 8125340 |
| analysis of three 2,3-dihydroxybiphenyl 1,2-dioxygenases found in rhodococcus globerulus p6. identification of a new family of extradiol dioxygenases. | the polychlorobiphenyl-degrading bacterium rhodococcus globerulus p6 contains three bphc genes encoding 2,3-dihydroxybiphenyl 1,2-dioxygenases. one of them, bphc1, is clustered with the bphb gene which encodes 2,3-dihydroxy-4-phenylhexa-4,6-diene dehydrogenase and constitutes part of the bph operon specifying the degradation of biphenyl. the nucleotide sequence of bphb and the three bphc genes has been determined. the protein products of the bphbc1 gene cluster were found to exhibit significant ... | 1994 | 8126007 |
| 13c nuclear magnetic resonance studies of pseudomonas putida fatty acid metabolic routes involved in poly(3-hydroxyalkanoate) synthesis. | the formation of poly(3-hydroxyalkanoates) (phas) in pseudomonas putida kt2442 from various carbon sources was studied by 13c nuclear magnetic resonance spectroscopy, gas chromatography, and gas chromatography-mass spectroscopy. by using [1-13c]decanoate, the relation between beta-oxidation and pha formation was confirmed. the labeling pattern in phas synthesized from [1-13c]acetate corresponded to the formation of phas via de novo fatty acid biosynthesis. studies with specific inhibitors of the ... | 1994 | 8132461 |
| 3-carboxy-cis,cis-muconate lactonizing enzyme from neurospora crassa: an alternate cycloisomerase motif. | 3-carboxy-cis,cis-muconate lactonizing enzyme (cmle; ec 5.5.1.5) from neurospora crassa catalyzes the reversible gamma-lactonization of 3-carboxy-cis,cis-muconate by a syn-1,2 addition-elimination reaction. the stereochemical and regiochemical course of the reaction is (i) opposite that of cmle from pseudomonas putida (ec 5.5.1.2) and (ii) identical to that of cis,cis-muconate lactonizing enzyme (mle; ec 5.5.1.1) from p. putida. in order to determine the mechanistic and evolutionary relationship ... | 1994 | 8132467 |
| carbon source-dependent inhibition of xyl operon expression of the pseudomonas putida tol plasmid. | tol plasmid-encoded degradation of benzyl alcohol by pseudomonas putida is inhibited by glucose and other compounds related to the main carbohydrate metabolism in pseudomonas species. we report here that this effect is exerted at the level of expression of the xyl catabolic operons, and two xyl promoters, pu and ps, were identified as the primary targets of this inhibition. xyl promoter activation was also inhibited by glucose in the heterologous escherichia coli system, apparently not however b ... | 1994 | 8132475 |
| genetic evidence for activation of the positive transcriptional regulator xy1r, a member of the ntrc family of regulators, by effector binding. | the xy1r protein positively controls expression from the pseudomonas putida tol plasmid sigma 54-dependent pu and ps promoters, in response to the presence of aromatic effectors such as m-xylene, m-methylbenzyl alcohol, and p-chlorobenzaldehyde in the culture medium. xy1r also autoregulates its own synthesis. a mutant xy1r regulator called xy1r7 was isolated after nitrosoguanidine mutagenesis of the wild-type gene and phenotypic selection for mutants that had acquired the ability to recognize m- ... | 1994 | 8132529 |
| regulation of the rpon, orf102 and orf154 genes in pseudomonas putida. | the dna sequence downstream of the pseudomonas putida rpon gene and the adjacent orf102 was determined. this region encodes an orf (orf154) whose gene product was found to be homologous to a family of phosphotransferases. insertional mutagenesis and analysis of mrna transcripts showed that the rpon gene is transcribed separately from the two downstream genes. the rpon promoter was localized to an 86 nucleotide-long region upstream of the rpon gene by examination of the expression of a series of ... | 1994 | 8138132 |
| efficient degradation of trichloroethylene by a hybrid aromatic ring dioxygenase. | engineering of hybrid gene clusters between the toluene metabolic tod operon and the biphenyl metabolic bph operon greatly enhanced the rate of biodegradation of trichloroethylene. escherichia coli cells carrying a hybrid gene cluster composed of todc1 (the gene encoding the large subunit of toluene terminal dioxygenase in pseudomonas putida f1), bpha2 (the gene encoding the small subunit of biphenyl terminal dioxygenase in pseudomonas pseudoalcaligenes kf707), bpha3 (the gene encoding ferredoxi ... | 1994 | 8144482 |
| molecular analysis of the erwinia chrysanthemi region containing the kdga and zwf genes. | the pathways of pectin and galacturonate catabolism in erwinia chrysanthemi converge to form a common intermediate, 2-keto-3-deoxygluconate, which is phosphorylated to form 2-keto-3-deoxy-6-phosphogluconate (kdgp) and then cleaved by the aldolase encoded by the kdga gene. we cloned the kdga gene of the e. chrysanthemi strain 3937 by complementing an escherichia coli kdga mutation, using an rp4-derivative plasmid. restriction mapping of the kdga region and isolation of kdga-lac fusions allowed th ... | 1994 | 8145647 |
| sequence and structure comparison suggest that methionine aminopeptidase, prolidase, aminopeptidase p, and creatinase share a common fold. | amino acid sequence comparison suggests that the structure of escherichia coli methionine aminopeptidase (ec 3.4.11.18) and the c-terminal domain of pseudomonas putida creatinase (ec 3.5.3.3) are related. a detailed comparison of the three-dimensional folds of the two enzymes confirms this homology: with an approximately 260-residue chain segment, 218 c alpha atoms of the structures superimpose within 2.5 a; only 41 of these overlapping positions (i.e., 19%) feature identical amino acids in the ... | 1994 | 8146141 |
| metabolism of naphthalene, fluorene, and phenanthrene: preliminary characterization of a cloned gene cluster from pseudomonas putida ncib 9816. | a modified cloning procedure was used to obtain large dna insertions (20 to 30 kb) from pseudomonas putida ncib 9816 that expressed polycyclic aromatic hydrocarbon (pah) transformation activity in escherichia coli hb101. four subclones (16 [in both orientations], 12, and 8.5 kb in size) were constructed from the initial clones. naphthalene, fluorene, and phenanthrene transformations were investigated in these eight ncib 9816 clones by a simple agar plate assay method, which was developed to dete ... | 1994 | 8157584 |
| inducibility of the tol catabolic pathway in pseudomonas putida (pww0) growing on succinate in continuous culture: evidence of carbon catabolite repression control. | the tol catabolic genes in pseudomonas putida (pww0) are clustered in the upper operon, encoding enzymes for the conversion of toluene and xylenes to benzoate and toluates, and the meta-cleavage operon, encoding enzymes for the conversion of the benzoate and toluates to tricarboxylic acid cycle intermediates. in this study, it was shown that cells growing in a chemostat under succinate growth-limiting conditions express both the upper and meta-cleavage pathways in response to o-xylene, a nonmeta ... | 1994 | 8157604 |
| cloning and characterization of a chromosomal gene cluster, pah, that encodes the upper pathway for phenanthrene and naphthalene utilization by pseudomonas putida ous82. | a 25-kb dna sali fragment cloned from the chromosomal dna of pseudomonas putida ous82, which utilizes phenanthrene (phn+) and naphthalene (nah+), carried all of the genes necessary for upper naphthalene catabolism. cosmid recombinant pip7 complemented both the nah- and phn- defects of ous8211 (trp-nah-phn-sal+[salicylate utilizing]hna+[1-hydroxy-2-naphthoate utilizing]) and only the phn- defect of ous8212 (trp-nah-phn-sal-hna+). the results indicate that strain ous82 uses different pathways afte ... | 1994 | 8157614 |
| identification and characterization of genes encoding polycyclic aromatic hydrocarbon dioxygenase and polycyclic aromatic hydrocarbon dihydrodiol dehydrogenase in pseudomonas putida ous82. | naphthalene and phenanthrene are transformed by enzymes encoded by the pah gene cluster of pseudomonas putida ous82. the paha and pahb genes, which encode the first and second enzymes, dioxygenase and cis-dihydrodiol dehydrogenase, respectively, were identified and sequenced. the dna sequences showed that paha and pahb were clustered and that paha consisted of four cistrons, pahaa, pahab, pahac, and pahad, which encode ferredoxin reductase, ferredoxin, and two subunits of the iron-sulfur protein ... | 1994 | 8157615 |
| identification and characterization of a transmissible linear plasmid from rhodococcus erythropolis bd2 that encodes isopropylbenzene and trichloroethene catabolism. | rhodococcus erythropolis bd2, which is able to utilize isopropylbenzene as a sole carbon and energy source, was shown to contain a conjugative linear plasmid, pbd2. the estimated size of pbd2 is 208 to 212 kb. linear plasmid-deficient strains had lost both the isopropylbenzene degradation and trichloroethene degradation characteristics, as well as the arsenite resistance and mercury resistance phenotypes. reintroduction of pbd2 restored all four characteristics. conjugational transfer of pbd2 to ... | 1994 | 8161179 |
| cloning, sequencing, and expression in escherichia coli of the d-hydantoinase gene from pseudomonas putida and distribution of homologous genes in other microorganisms. | pseudomonas putida dsm 84 produces n-carbamyl-d-amino acids from the corresponding d-5-monosubstituted hydantoins. the gene encoding this d-hydantoinase enzyme was cloned and expressed in escherichia coli. the nucleotide sequence of the 1.8-kb insert of subclone pges19 was determined. one open reading frame of 1,104 bp was found and was predicted to encode a polypeptide with a molecular size of 40.5 kda. local regions of identity between the predicted amino acid sequence and that of other known ... | 1994 | 8161181 |
| transfer and expression of degradative and antibiotic resistance plasmids in acidophilic bacteria. | the genetic accessibility of selected acidophilic bacteria was investigated to evaluate their applicability to degrading pollutants in acidic environments. the incp1 antibiotic resistance plasmids rp4 and pvk101 and the phenol degradation-encoding plasmid ppgh11 were transferred from neutrophilic bacteria into the extreme acidophilic eubacterium acidiphilium cryptum at frequencies of 1.8 x 10(-2) to 9.8 x 10(-4) transconjugants per recipient cell. the incq antibiotic resistance plasmid psup106 w ... | 1994 | 8161188 |
| aliphatic and aromatic inhibitors binding to the active site of catechol 2,3-dioxygenase from pseudomonas putida mt-2. | the interaction of different classes of inhibitors with the extradiol cleaving catechol 2,3-dioxygenase from pseudomonas putida mt-2 was monitored by longitudinal and transverse proton relaxation measurements as well as by kinetic activity studies in order to characterize the type of interaction of such inhibitors with the active site of the enzyme. the average distances of the inhibitors from the catalytic iron(ii) ion have been estimated from the 1h longitudinal relaxation rates. phenols and a ... | 1994 | 8163017 |
| studies on the oxidative half-reaction of p-hydroxyphenylacetate 3-hydroxylase. | the oxidative half-reaction of the two-protein enzyme, p-hydroxyphenylacetate 3-hydroxylase from pseudomonas putida, has been studied by absorbance stopped-flow techniques. the formation of three flavin-oxygen intermediates, the anionic and protonated forms of the flavin hydroperoxide (intermediates i and i) and the hydroxyflavin (intermediate iii), was observed during the course of the oxygen reaction with the reduced flavoprotein-coupling protein complex. the flavin hydroperoxide, which is for ... | 1994 | 8163477 |
| engineering enzymes for clinical diagnosis. | the enzyme creatine amidinohydrolase (creatinase, ec 3.5.3.3.) is a major limiting factor in the enzymatic creatinine determination because of its comparatively poor catalytic activity and stability in the native form. the gene from pseudomonas putida coding for creatinase was cloned and used for overexpression of the protein in e coli and pseudomonas. in addition, it was possible by means of 'random' mutagenesis in vivo and subsequent screening using an activity plate assay to isolate creatinas ... | 1993 | 8166396 |
| catabolism of aromatics in pseudomonas putida u. formal evidence that phenylacetic acid and 4-hydroxyphenylacetic acid are catabolized by two unrelated pathways. | phenylacetic acid (phacoh) and 4-hydroxyphenylacetic acid (4hophacoh) are catabolized in pseudomonas putida u through two different pathways. mutation carried out with the transposon tn5 has allowed the isolation of several mutants which, unlike the parental strain, are unable to grow in chemically defined medium containing either phacoh or 4hophacoh as the sole carbon source. analysis of these strains showed that the ten mutants unable to grow in phacoh medium grew well in the one containing 4h ... | 1994 | 8168524 |
| cloning and high-level expression of the glutathione-independent formaldehyde dehydrogenase gene from pseudomonas putida. | a dna fragment of 485 bp was specifically amplified by pcr with primers based on the n-terminal sequence of the purified formaldehyde dehydrogenase (ec 1.2.1.46) from pseudomonas putida and on that of a cyanogen bromide-derived peptide. with this product as a probe, a gene coding for formaldehyde dehydrogenase (fdha) in p. putida chromosomal dna was cloned in escherichia coli dh5 alpha. sequencing analysis revealed that the fdha gene contained 1,197-bp open reading frame, encoding a protein comp ... | 1994 | 8169197 |
| the effect of ferredoxin(bed) overexpression on benzene dioxygenase activity in pseudomonas putida ml2. | the benzene dioxygenase from pseudomonas putida ml2 is a multicomponent complex comprising a flavoprotein reductase, a ferredoxin, and a terminal iron-sulfur protein (isp). the catalytic activity of the isolated complex shows a nonlinear relationship with protein concentration in cell extracts, with the limiting factor for activity in vitro being ferredoxin(bed). the relative levels of the three components were analyzed by using 125i-labelled antibodies, and the functional molar ratio of isp(bed ... | 1994 | 8169199 |
| transcriptional induction kinetics from the promoters of the catabolic pathways of tol plasmid pww0 of pseudomonas putida for metabolism of aromatics. | we determined, under several growth conditions, the kinetics of mrna synthesis from the four pseudomonas putida pww0 plasmid promoters involved in degradation of xylenes and methylbenzyl alcohols via toluates. transcription by xyls of the meta-cleavage pathway operon promoter (pm) for the metabolism of alkylbenzoates was stimulated immediately after the addition of an effector, both in luria-bertani (lb) medium and in minimal medium. activation of the sigma 54-dependent upper-pathway operon prom ... | 1994 | 8169200 |
| secondary 18o and primary 13c isotope effects as a probe of transition-state structure for enzymatic decarboxylation of oxalacetate. | a new method for directly measuring 18o isotope effects on decarboxylation reactions has been developed. by running the reaction under high vacuum (10(-5) torr), co2 leaves the solution before exchange with the oxygens of water to an extent greater than 2%. thus, the method permits determination of 18o isotope effects with the precision of the isotope ratio mass spectrometer, and without the necessity of resorting to the remote label method and its attendant required syntheses. the method is use ... | 1994 | 8172901 |
| molecular characterization of microbial alcohol dehydrogenases. | there is an astonishing array of microbial alcohol oxidoreductases. they display a wide variety of substrate specificities and they fulfill several vital but quite different physiological functions. some of these enzymes are involved in the production of alcoholic beverages and of industrial solvents, others are important in the production of vinegar, and still others participate in the degradation of naturally occurring and xenobiotic aromatic compounds as well as in the growth of bacteria and ... | 1994 | 8185833 |
| bacterial morphine dehydrogenase further defines a distinct superfamily of oxidoreductases with diverse functional activities. | pseudomonas putida morphine dehydrogenase is shown to be closely homologous to 18 proteins, defining a superfamily within which morphine dehydrogenase particularly resembles two bacterial, 2,5-dioxo-d-gluconic acid reductases, and two eukaryotic proteins of unknown functions. relationships within the superfamily are extensive and complex. residue identities between protein pairs range from 29-90%. three subgroups are proposed. nevertheless, on the basis of residue conservations/exchanges it is s ... | 1994 | 8192670 |
| genetic evidence that the xyls regulator of the pseudomonas tol meta operon controls the pm promoter through weak dna-protein interactions. | the activation of the pm promoter of the meta operon of the tol plasmid of pseudomonas putida by its cognate xyls activator protein in the presence and absence of benzoate inducers has been examined in specialized escherichia coli strains carrying pm-lacz fusions and the xyls gene in different configurations in which all controlling elements are present in near native conditions and stoichometry. expression of a chromosomal pm-xylx::lacz fusion was primarily dependent on the addition of an effec ... | 1994 | 8195070 |
| oxidation of low molecular weight chloroalkanes by cytochrome p450cam. | cytochrome p450cam from pseudomonas putida g786 oxidized chlorinated ethanes and 1,2-dichloropropane. the rate of nadh oxidation decreased with decreasing chlorine substitution. the single detectable oxidation products of 1,1,1-trichloroethane and 1,2-dichloropropane were 1,1,1-trichloroethanol and chloroacetone, respectively. organic product formation was largely uncoupled from nadh oxidation. | 1994 | 8198597 |
| identification of serine-143 as the most likely precursor of dehydroalanine in the active site of histidine ammonia-lyase. a study of the overexpressed enzyme by site-directed mutagenesis. | the gene coding for histidase (histidine ammonia-lyase, hal, ec 4.3.1.3) was isolated from a lambda-embl3 genomic library from pseudomonas putida nicii and subcloned into the expression vector pt7-7. transformation of escherichia coli bl21 (de3) cells with the recombinant vector led to the expression of catalytically active histidase amounting to 20-30% of the total soluble protein in the crude cell extract. a new rapid and highly efficient isolation procedure is described leading to electrophor ... | 1994 | 8204579 |
| variation in chlorobenzoate catabolism by pseudomonas putida p111 as a consequence of genetic alterations. | pseudomonas putida p111 is able to utilize a broad range of monochlorinated, dichlorinated, and trichlorinated benzoates. the involvement of two separate dioxygenases was noted from data on plasmid profiles and dna hybridization. the benzoate dioxygenase, which converts 3-chlorobenzoate (3-cb), 4-cb, and benzoate to the corresponding catechols via reduction of a dihydrodiol, was shown to be chromosomally coded. the chlorobenzoate-1,2-dioxygenase that converts ortho-chlorobenzoates to the corresp ... | 1993 | 8215353 |
| 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 |
| 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 |
| 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 |
| the oct plasmid encodes d-lysine membrane transport and catabolic enzymes in pseudomonas putida. | pseudomonas putida (oleovorans) (pp(oct)) cured of its oct plasmid (pp) no longer grows on d-lysine. conjugation of pptrp- with three different methionine auxotrophs carrying the oct plasmid resulted in pptrp- (oct) organisms that grew on d-lysine. three early d-lysine catabolic enzymes encoded by the oct plasmid are a lysine racemase, the proposed conversion of d-lysine to delta 1-piperidine-2-carboxylate (p2c), for which we provide evidence, and p2c reductase which converts p2c to pipecolate. ... | 1993 | 8234494 |
| depletion of serum methionine by methioninase in mice. | methionine dependence is a tumor-specific metabolic defect found in human cancer cell lines as well as in fresh human tumor specimens. methionine dependent tumors cease growing when deprived of methionine, unlike normal cells which can substitute homocysteine for methionine for their growth requirement. we have previously purified a stable, endotoxin-free methioninase from the bacterium, pseudomonas putida. we demonstrate in this report that purified methioninase can lower the serum levels of me ... | 1993 | 8239522 |
| identification of ser143 as the site of modification in the active site of histidine ammonia-lyase. | histidine ammonia-lyase (histidase) from pseudomonas putida was irreversibly inactivated by l-cysteine at ph 10.5 in the presence of oxygen. inactivation was accompanied by the formation of a new uv-absorbing species centered around 340 nm. l-[35s]cysteine labeling experiments revealed that 4 mol of l-cysteine was bound per mole of enzyme tetramer upon complete modification. however, the radiolabel was dissociated from the protein under denaturing conditions without loss of the 340-nm absorbance ... | 1993 | 8239649 |
| 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 |