Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
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| 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 |
| 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 |
| 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 |
| 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 |
| adaptive mutation: the uses of adversity. | when populations of microorganisms are subjected to certain nonlethal selections, useful mutants arise among the nongrowing cells whereas useless mutants do not. this phenomenon, known as adaptive, directed, or selection-induced mutation, challenges the long-held belief that mutations only arise at random and without regard for utility. in recent years a growing number of studies have examined adaptive mutation in both bacteria and yeast. although conflicts and controversies remain, the weight o ... | 1993 | 8257106 |
| essential role of the arg112 residue of cytochrome p450cam for electron transfer from reduced putidaredoxin. | cytochrome p450cam (cyp101) of pseudomonas putida ppg1 in which arg112 is substituted by cys was isolated by in vitro random mutagenesis of the camc gene dna coding for p450cam. the absorption spectra of the purified mutant enzyme were similar to those of the wild type enzyme, but its substrate-dependent nadh oxidation activity in the presence of putidaredoxin (pd) and putidaredoxin reductase (pdr) was extremely low. the rate constant of electron transfer from reduced pd to the heme of the mutan ... | 1993 | 8405387 |
| a mutagenesis system utilizing a tn1722 derivative containing an escherichia coli-specific vector plasmid: application to pseudomonas species. | a novel transposon (tn) mutagenesis system for gram- non-enteric bacteria was developed which allowed rapid and one-step cloning of the mutated region in escherichia coli. the tn constructed was tn1722-299km, a tn1722 derivative containing a kmr gene and the entire sequence of an e. coli-specific plasmid, pacyc184. the hybrid plasmid consisting of tn1722-299km and the transfer genes of plasmid r388 was conjugally transferred from e. coli to pseudomonas putida or p. aeruginosa, and selection of t ... | 1993 | 8294012 |
| [the role of pyrimidines in the biosynthesis of the fluorescing pigment pyoverdin pm in pseudomonas putida m]. | dihydroorotate was shown to be a predecessor of deoxyquinoline nucleus of a fluorescing enzyme pyoverdin pm in pseudomonas putida m. the data was obtained in experiments with a set of pyr- mutants with different steps of pyrimidine synthesis blocked. a scheme for deoxyquinoline nucleus of the enzyme including dihydroorotate participation is proposed. | 1993 | 8289842 |
| construction of chromosomal reca mutants of pseudomonas putida ppg2. | the reca gene of pseudomonas putida ppg2 was cloned by complementation of the reca mutations of escherichia coli strains dh5 alpha and hb101. the nucleotide sequence of the dna fragment was determined and shown to contain reca and a downstream partial open reading frame. two mutants of p. putida ppg2, strains js387 and js388, were constructed by insertional inactivation of reca with a tetracycline-resistance gene in both orientations. both mutants acquired sensitivity to methyl methanesulfonate ... | 1993 | 8294013 |
| analysis of the dna damage-mediated induction of pseudomonas putida and pseudomonas aeruginosa lexa genes. | a fusion between the lexa gene of pseudomonas aeruginosa and pseudomonas putida and the lacz gene was constructed in vitro and cloned in a mini-tn5 transposon derivative to obtain chromosomal insertions which enable to quantitatively examine their transcriptional regulation in both pseudomonas and e. coli. analysis of dna damage-mediated induction of these lexa-lacz fusions showed that expression of p. putida and p. aeruginosa lexa genes was always higher and earlier than the expression of the l ... | 1993 | 8319897 |
| the bkdr gene of pseudomonas putida is required for expression of the bkd operon and encodes a protein related to lrp of escherichia coli. | branched-chain keto acid dehydrogenase is a multienzyme complex which is required for the metabolism of the branched-chain amino acids in pseudomonas putida. the structural genes encoding all four proteins of the bkd operon have been cloned, and their nucleotide sequences have been determined (g. burns, k. t. madhusudhan, k. hatter, and j. r. sokatch, p. 177-184 in s. silver, a. m. chakrabarty, b. iglewski, and s. kaplan [ed.], pseudomonas: biotransformations, pathogenesis, and evolving biotechn ... | 1993 | 8320210 |
| a new amino acid racemase with threonine alpha-epimerase activity from pseudomonas putida: purification and characterization. | we have found that pseudomonas putida atcc 17642 cells grown in a medium containing d-threonine as the sole nitrogen source produce an enzyme that catalyzes epimerization of threonine. proton nuclear magnetic resonance analysis of the enzyme reaction in deuterium oxide clearly showed epimerization from l- to d-allo-threonine and also from d- to l-allo-threonine. this is the first example of an enzyme that was clearly shown to epimerize threonine. the enzyme has been purified to homogeneity, whic ... | 1993 | 8320235 |
| expression of the agga locus of pseudomonas putida in vitro and in planta as detected by the reporter gene, xyle. | in vitro agglutinability by pseudomonas putida, isolate corvallis, with a plant root surface agglutinin is correlated with rapid adhesion of cells of the fluorescent pseudomonad to bean (phaseolus vulgaris) root surfaces. agglutinability in p. putida cells is regulated by nutrient status as well as growth phase. cells grown in three different nutrient complex media are agglutinable at early and mid-late logarithmic phase but become nonagglutinable at stationary phase. cells grown in a minimal me ... | 1993 | 8324250 |
| cloning and nucleotide sequence analysis of the ferripyoverdine receptor gene fpva of pseudomonas aeruginosa. | pseudomonas aeruginosa k437 lacks the ferripyoverdine receptor and, as a result, grows poorly on an iron-deficient minimal medium supplemented with ethylenediamine-di(o-hydroxyphenylacetic acid) (eddha) and pyroverdine. by using a phagemid-based in vivo cloning system, attempts were made to clone the receptor gene by complementing this growth defect. several recombinant phagemids carrying p. aeruginosa chromosomal dna which provided for good growth on eddha-pyoverdine-containing medium and which ... | 1993 | 8335619 |
| nucleotide sequence and over-expression of morphine dehydrogenase, a plasmid-encoded gene from pseudomonas putida m10. | pseudomonas putida m10 was originally isolated from factory waste liquors by selection for growth on morphine. the nadp(+)-dependent morphine dehydrogenase that initiates morphine catabolism is encoded by a large plasmid of 165 kb. treatment of p. putida m10 with ethidium bromide led to the isolation of a putative plasmid-free strain that was incapable of growth on morphine. the structural gene for morphine dehydrogenase, mora, has been located on the plasmid by oligonucleotide hybridization, by ... | 1993 | 8452544 |
| the pseudomonas putida ml2 plasmid-encoded genes for benzene dioxygenase are unusual in codon usage and low in g+c content. | benzene dioxygenase, catalyzing the oxidation of benzene to cis-1,2-dihydroxy-cyclohexa-3,5-diene, comprises four polypeptides that are encoded by plasmid phmt112 of pseudomonas putida ml2. in this study, the nucleotide (nt) sequences of four genes encoding this enzyme (bedc1c2ba) were determined, and the amino acid (aa) sequences were deduced. the sequence showed significant homology with the chromosomally encoded benzene dioxygenase and toluene dioxygenase genes (73-77% for nt and 83-99% for a ... | 1993 | 8344526 |
| degradative capability of pseudomonas putida on acetonitrile. | pseudomonas putida, capable of utilizing acetonitrile as a sole source of carbon and nitrogen, was isolated from contaminated soil and water samples collected from industrial sites. the p. putida cells were immobilized in calcium alginate beads. the degradation of acetonitrile by the immobilized cells of p. putida was investigated. the immobilized cells degraded different concentrations of acetonitrile into ammonia and carbon dioxide. the effect of aeration on the degradation rate was also studi ... | 1993 | 8323268 |
| plasmid-mediated mineralization of naphthalene, phenanthrene, and anthracene. | the well-characterized plasmid-encoded naphthalene degradation pathway in pseudomonas putida ppg7(nah7) was used to investigate the role of the nah plasmid-encoded pathway in mineralizing phenanthrene and anthracene. three pseudomonas strains, designated 5r, dfc49, and dfc50, were recovered from a polynuclear aromatic hydrocarbon-degrading inoculum developed from a manufactured gas plant soil slurry reactor. plasmids pka1, pka2, and pka3, approximately 100 kb in size, were isolated from these st ... | 1993 | 8328809 |
| in vivo reactivation of catechol 2,3-dioxygenase mediated by a chloroplast-type ferredoxin: a bacterial strategy to expand the substrate specificity of aromatic degradative pathways. | the meta-cleavage operon of the tol plasmid pww0 of pseudomonas putida contains 13 genes responsible for the oxidation of benzoate and toluates to krebs cycle intermediates via estradiol (meta) cleavage of (methyl)catechol. the functions of all the genes are known with the exception of xylt. we constructed pww0 mutants defective in the xylt gene, and found that these mutants were not able to grow on p-toluate while they were still capable of growing on benzoate and m-toluate. in the xylt mutants ... | 1993 | 8344270 |
| gene components responsible for discrete substrate specificity in the metabolism of biphenyl (bph operon) and toluene (tod operon). | bph operons coding for biphenyl-polychlorinated biphenyl degradation in pseudomonas pseudoalcaligenes kf707 and pseudomonas putida kf715 and tod operons coding for toluene-benzene metabolism in p. putida f1 are very similar in gene organization as well as size and homology of the corresponding enzymes (g. j. zylstra and d. t. gibson, j. biol. chem. 264:14940-14946, 1989; k. taira, j. hirose, s. hayashida, and k. furukawa, j. biol. chem. 267:4844-4853, 1992), despite their discrete substrate rang ... | 1993 | 8349562 |
| identification of a cis-acting sequence within the pm promoter of the tol plasmid which confers xyls-mediated responsiveness to substituted benzoates. | the dna sequences within the pm promoter/operator region of the meta operon of the tol plasmid of pseudomonas putida, which confer xyls-mediated responsiveness to substituted benzoates, have been identified through deletion analysis and mutagenesis of the region. integrity and proper phasing of two homologous tandem sequences 5'-tgcaapuaapu-pyggnta-3', separated by six base-pairs and overlapping with the -35 region of the pm promoter, was essential for m-toluate activation of a pm-lacz fusion in ... | 1993 | 8478926 |
| indirect utilization of the phytosiderophore mugineic acid as an iron source to rhizosphere fluorescent pseudomonas. | the phytosiderophore mugineic acid (ma) was studied as a source of iron for rhizosphere fluorescent pseudomonads. 55fe supplied as fe-ma was taken up by pseudomonas putida wcs358, b10 and st3 grown under iron deficient conditions. the uptake decreased when the bacteria were grown in the presence of iron. however, no differences in uptake were observed when a siderophore deficient mutant was tested. since ligand exchange between pseudobactin and ma was shown to occur rapidly with a half-life of 2 ... | 1993 | 8358206 |
| regulation of the pcaij genes for aromatic acid degradation in pseudomonas putida. | six of the genes encoding enzymes of the beta-ketoadipate pathway for benzoate and 4-hydroxybenzoate degradation in pseudomonas putida are organized into at least three separate transcriptional units. as an initial step to defining this pca regulon at the molecular level, lacz fusions were made with the pcai and pcaj genes, which encode the two subunits of beta-ketoadipate:succinyl-coenzyme a transferase, the enzyme catalyzing the next-to-last step in the beta-ketoadipate pathway. fusion analyse ... | 1993 | 8376330 |
| cbbr, a lysr-type transcriptional activator, is required for expression of the autotrophic co2 fixation enzymes of xanthobacter flavus. | xanthobacter flavus is able to grow autotrophically with the enzymes of the calvin cycle for the fixation of co2, which are specified by the cbblsxfp gene cluster. previously, the 5' end of an open reading frame (cbbr), displaying a high sequence similarity to the lysr family of regulatory proteins and transcribed divergently from cbblsxfp, was identified (w. g. meijer, a. c. arnberg, h. g. enequist, p. terpstra, m. e. lidstrom, and l. dijkhuizen, mol. gen. genet. 225:320-330, 1991). this paper ... | 1993 | 8407781 |
| cloning and expression of a member of a new cytochrome p-450 family: cytochrome p-450lin (cyp111) from pseudomonas incognita. | cytochrome p-450lin catalyzes the 8-methyl hydroxylation of linalool as the first committed step of its utilization by pseudomonas incognita as the sole carbon source. by using a polymerase chain reaction-based cloning strategy, a 2.1-kb dna fragment containing the cytochrome p-450lin gene (linc) was isolated. an open reading frame of 406 amino acids has been identified as that of p-450lin on the basis of amino acid sequence data from peptides of the native protein. heterologous expression of fu ... | 1993 | 8376348 |
| energy conservation by pyrroloquinoline quinol-linked xylose oxidation in pseudomonas putida nctc 10936 during carbon-limited growth in chemostat culture. | when grown in carbon source-limited chemostat cultures with lactate or glucose as the carbon and energy source and xylose as an additional source of reducing equivalents. pseudomonas putida nctc 10936 oxidized xylose to xylonolactone and xylonate. no other products were formed from this pentose, nor was it incorporated into biomass. the presence of xylose in these cultures resulted in higher yglucose and ylactate values as compared to cultures without xylose indicating that biologically useful e ... | 1993 | 8385642 |
| in-vivo-generated fusion promoters in pseudomonas putida. | plasmid pest1463 carrying the promoterless pheba operon was cloned into pseudomonas putida paw85, and phenol-utilizing colonies were isolated on minimal plates containing phenol as the only carbon and energy source. in these clones, chromosomally located tn4652 was transposed upstream from the coding sequencing of phea (encoding phenol monooxygenase). sequence analysis together with mapping of the transcription start point of the pheba operon in the recombinant plasmids revealed that fusions of ... | 1993 | 8387446 |
| identification and characterization of the pupb gene encoding an inducible ferric-pseudobactin receptor of pseudomonas putida wcs358. | pseudomonas putida wcs358 can transport iron complexed to a wide variety of pseudobactins produced by other pseudomonas strains. the pupb gene encoding an outer membrane ferric-pseudobactin receptor was isolated from a genomic library of p. putida wcs358. the pupb receptor facilitated iron transport via two distinct heterologous siderophores, i.e. pseudobactin bn8 and pseudobactin bn7. the amino acid sequence deduced from the nucleotide sequence consisted of 804 amino acids (molecular weight 88, ... | 1993 | 8392140 |
| engineering of alkyl- and haloaromatic-responsive gene expression with mini-transposons containing regulated promoters of biodegradative pathways of pseudomonas. | four recombinant mini-tn5 transposons are described which contain outward-facing pm, pu or psal promoters from the catabolic plasmids tol and nah of pseudomonas putida, along with their cognate wild-type regulatory genes (xyls, xylr, nahr) or mutant varieties (xyls2). transcription from such promoters is activated when the host bacteria encounters certain aromatic compounds, such as alkyl- and halobenzoates (xyls, xyls2), alkyl- and halotoluenes (xylr) or salicylates (nahr). these transposons en ... | 1993 | 8393826 |
| reinvestigation of the role of thiol groups of glyoxalase i purified from yeast (saccharomyces cerevisiae). | glyoxalase i has been purified to homogeneity from saccharomyces cerevisiae and tested with two different thiol reagents, i.e., 5,5'-dithiobis-(2-nitrobenzoic acid) (dtnb) and 1-chloro-2,4-dinitrobenzene (cdnb). dtnb reacts with four thiol groups per molecule of enzyme and leads to a complete inhibition which is not reversed by addition of the disulfide-reducing agent dithiothreitol. on the other hand, cdnb slightly affects the glyoxalase-i activity and alkylates only one thiol residue/enzyme. i ... | 1993 | 8373819 |
| transformation of 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8- dihydroxyquinoline by quinoline-grown resting cells of pseudomonas putida 86. | resting cells of pseudomonas putida strain 86 were grown on quinoline transformed 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8-dihydroxyquinoline which was not converted further. 7,8-dioxygenating activity was present when the enzymes of quinoline catabolism were induced. quinoline-grown cells of strain 86 treated simultaneously with 2-chloroquinoline and d-(-)-threo-chloramphenicol to prevent protein biosynthesis also formed the cis-7,8-dihydrodiol of 2-chloroquinoline. succinate-grown res ... | 1993 | 8405957 |
| heteronuclear nmr analysis of unsaturated fatty acids in poly(3-hydroxyalkanoates). study of beta-oxidation in pseudomonas putida. | poly(3-hydroxyalkanoates) (phas) were isolated from pseudomonas putida kt2442 cultivated on petroselenic acid, oleic acid, and linoleic acid to study beta-oxidation of unsaturated fatty acids. both saturated and unsaturated medium chain length 3-hydroxy fatty acids were found to be constituents of these polymers. with the aid of proton-detected multiple quantum coherence and proton-detected multiple bond coherence nmr spectra the structures of the unsaturated monomers were identified as 3-hydrox ... | 1993 | 8416939 |
| preelectrophoresis of agarose plugs containing bacterial chromosomal dna prepared for analysis by pulsed field gel electrophoresis can improve the clarity of restriction patterns. | pulsed field gel electrophoresis has indicated that chromosomal dna isolated from stationary phase pseudomonas fluorescens, pseudomonas putida, and escherichia coli cells immobilized in agarose can be fragmented during its release. for p. fluorescens it was demonstrated that the entire chromosome is affected and that there are no specifically fragile sites. the extent of the damage increased both during storage of the dna and also when magnesium ions were provided, suggesting that nucleases may ... | 1993 | 8387734 |
| cloning, sequencing, and genetic characterization of regulatory genes, rina and rinb, required for the activation of staphylococcal phage phi 11 int expression. | the int gene of staphylococcal bacteriophage phi 11 is the only viral gene responsible for the integrative recombination of phi 11. to study the regulation of int gene expression, we determined the 5' end of the transcript by s1 mapping. the presumed promoter is located just 22 nucleotides upstream of the int open reading frame in a region which is conserved between phi 11 and a closely related staphylococcal phage, l54a. to clone the possible regulatory gene, a vector which contained the report ... | 1993 | 8432703 |
| nucleotide sequence and initial functional characterization of the clcr gene encoding a lysr family activator of the clcabd chlorocatechol operon in pseudomonas putida. | the 3-chlorocatechol operon clcabd is central to the biodegradative pathway of 3-chlorobenzoate. the clcr regulatory gene, which activates the clcabd operon, was cloned from the region immediately upstream of the operon and was shown to complement an insertion mutation for growth on 3-chlorobenzoate. clcr activated the clca promoter, which controls expression of the clcabd operon, in trans by 14-fold in an in vivo promoter probe assay in pseudomonas putida when cells were incubated with 15 mm 3- ... | 1993 | 8419291 |
| identification and characterization of the exbb, exbd and tonb genes of pseudomonas putida wcs358: their involvement in ferric-pseudobactin transport. | catechol-cephalosporins are siderophore-like antibiotics which are taken up by cells of pseudomonas putida wcs358 via the ferric-siderophore transport pathway. mutants of strain wcs358 were isolated that are resistant to high concentrations of these antibiotics. these mutants failed to grow under iron-limiting conditions, and could not utilize different ferric-siderophores. the mutants fall in three complementation groups. the nucleotide sequence determination identified three contiguous open re ... | 1993 | 8437515 |
| precise deletions in large bacterial genomes by vector-mediated excision (vex). the trfa gene of promiscuous plasmid rk2 is essential for replication in several gram-negative hosts. | we have developed a simple and efficient method of vector-mediated excision (vex) for in vivo generation of precisely defined deletions in large bacterial genomes. the system uses homologous recombination with small cloned fragments on specialized pvex plasmids to insert directly repeated bacteriophage p1 loxp sites at positions flanking the region to be deleted. in the presence of cre recombinase, the loxp sites are efficiently recombined to produce the deletion. deletion endpoints can be direc ... | 1993 | 8450534 |
| cloning, sequencing, and expression of the pseudomonas putida protocatechuate 3,4-dioxygenase genes. | the genes that encode the alpha and beta subunits of protocatechuate 3,4-dioxygenase (3,4-pcd [ec 1.13.11.3]) were cloned from a pseudomonas putida (formerly p. aeruginosa) (atcc 23975) genomic library prepared in lambda phage. plaques were screened by hybridization with degenerate oligonucleotides designed using known amino acid sequences. a 1.5-kb smai fragment from a 15-kb primary clone was subcloned, sequenced, and shown to contain two successive open reading frames, designated pcah and pcag ... | 1993 | 8407791 |
| proteins induced by sulfate limitation in escherichia coli, pseudomonas putida, or staphylococcus aureus. | two-dimensional gel electrophoresis of proteins from escherichia coli, pseudomonas putida, and staphylococcus aureus, grown with methionine or one of a variety of organosulfates and organosulfonates as the sole source of sulfur, showed expression of specific sets of 7 to 14 proteins which were not observed during growth with sulfate or cysteine for all three species or with thiocyanate for p. putida and s. aureus. under the same conditions, arylsulfatase activity in p. putida and s. aureus was s ... | 1993 | 8432711 |
| construction of a pseudomonas hybrid strain that mineralizes 2,4,6-trinitrotoluene. | a bacterium, pseudomonas sp. strain c1s1, able to grow on 2,4,6-trinitrotoluene (tnt), 2,4- and 2,6-dinitrotoluene, and 2-nitrotoluene as n sources, was isolated. the bacterium grew at 30 degrees c with fructose as a c source and accumulated nitrite. through batch culture enrichment, we isolated a derivative strain, called pseudomonas sp. clone a, which grew faster on tnt and did not accumulate nitrite in the culture medium. use of tnt by these two strains as an n source involved the successive ... | 1993 | 8468288 |
| increased expression of the plasmid-determined 2,3-dihydroxybiphenyl dioxygenase gene in strains of escherichia coli, pseudomonas putida and pseudomonas aeruginosa. | a 6.5-kb ecori fragment containing the gene encoding 2,3-dihydroxybiphenyl dioxygenase from the plasmid pbs312 was cloned into broad host range plasmid rsf1010 and expressed in escherichia coli, pseudomonas putida and pseudomonas aeruginosa strains. the increased expression of the gene was orientation-dependent and probably due to the transcription read through from the streptomycin promoter of the vector. subcloning experiments of the psti fragments of pbs312 plasmid using vector pbr322 reveale ... | 1993 | 8454186 |
| sequences of genes encoding naphthalene dioxygenase in pseudomonas putida strains g7 and ncib 9816-4. | the multicomponent enzyme, naphthalene dioxygenase, initiates the metabolism of naphthalene by pseudomonas putida strains g7 (ppg7) and ncib 9816-4 (pp9816-4). the genes involved (nahaaabacad) are encoded by the nah7 and pdtg1 plasmids, respectively, and form part of the nah operon. the locations of the structural genes were determined on previously cloned fragments of dna. the nucleotide (nt) sequences were determined for nahaaab from pp9816-4 and for nahaaabacad from ppg7. the appropriate open ... | 1993 | 8486285 |
| the amino acid sequence of pseudomonas putida azurin. | the low molecular weight "blue" copper protein, azurin, has been purified from pseudomonas putida (ncib 9869) to homogeneity using various chromatographic techniques including reverse-phase hplc. the amino acid sequence of the n-terminus of the reduced and carboxymethylated protein yielded a single sequence corresponding to aeckv. the complete sequence, comprising 128 amino acid residues with a c-terminal sequence corresponding to tvtlk, was determined from the peptides obtained from a staphyloc ... | 1993 | 8489263 |
| enhanced mineralization of polychlorinated biphenyls in soil inoculated with chlorobenzoate-degrading bacteria. | an altamont soil containing no measurable population of chlorobenzoate utilizers was examined for the potential to enhance polychlorinated biphenyl (pcb) mineralization by inoculation with chlorobenzoate utilizers, a biphenyl utilizer, combinations of the two physiological types, and chlorobiphenyl-mineralizing transconjugants. biphenyl was added to all soils, and biodegradation of 14c-aroclor 1242 was assessed by disappearance of that substance and by production of 14co2. mineralization of pcbs ... | 1993 | 8476293 |
| oxidation of aniline to nitrobenzene by nonheme bromoperoxidase. | nonheme bromoperoxidase found in pseudomonas putida catalyzed the bromination of aniline with hydrogen peroxide and bromide ions to give o- and p-bromoanilines. however, in the absence of bromide ions, it oxidized aniline via azobenzene and azoxybenzene finally into nitrobenzene. this is the first report of the biological oxidation of an arylamine to the corresponding nitrocompound at the enzyme level. in addition, nitrobenzene was not formed by a nonheme bromoperoxidase from corallina pilulifer ... | 1993 | 8490583 |
| kinetic studies on benzyl alcohol dehydrogenase encoded by tol plasmid pwwo. a member of the zinc-containing long chain alcohol dehydrogenase family. | the nucleotide sequence of the structural gene for benzyl alcohol dehydrogenase encoded by tol plasmid pwwo of pseudomonas putida has been determined. benzyl alcohol dehydrogenase is a member of the long-chain zinc alcohol dehydrogenase family and, like other alcohol dehydrogenases of this family, contains two zinc atoms per subunit. benzyl alcohol dehydrogenase, while sharing 31% identical residues with horse liver alcohol dehydrogenase, contains several amino acid substitutions near the active ... | 1993 | 8496150 |
| copper accumulation by a strain of pseudomonas putida. | a study on the copper accumulation by resting cells of copper-resistant bacteria, isolated from activated sludge and electroplating effluent, was conducted. the best selected strain, identified as pseudomonas putida ii-11, retained copper ions, cu(ii), as high as 6.5% of its dry weight. bacterial cells grown in the sulphate-limiting medium had the highest copper removal capacity [rc, mg of cu(ii)/g of dry cells], while the presence of glucose or sodium azide did not affect cu(ii) rc of the bacte ... | 1993 | 8459779 |
| isolation and preliminary characterization of the subunits of the terminal component of naphthalene dioxygenase from pseudomonas putida ncib 9816-4. | the terminal oxygenase component (ispnap) of naphthalene dioxygenase from pseudomonas putida ncib 9816-4 was purified to homogeneity. the protein contained approximately 4 g-atoms each of iron and acid-labile sulfide per mol of ispnap, and enzyme activity was stimulated significantly by addition of exogenous iron. the large (alpha) and small (beta) subunits of ispnap were isolated by two different procedures. the nh2-terminal amino acid sequences of the alpha and beta subunits were identical to ... | 1993 | 8376335 |
| molecular analysis of the plasmid-borne bed gene cluster from pseudomonas putida ml2 and cloning of the cis-benzene dihydrodiol dehydrogenase gene. | pseudomonas putida ml2 contains a large catabolic plasmid, phmt112, carrying genes that encode the dioxygenase and dehydrogenase involved in the catabolism of benzene via the ortho or beta-ketoadipate pathway. phmt112 was derived from a larger and less stable plasmid in p. putida ml2 following growth on succinate as carbon and energy source but was, however, stably maintained in p. putida even in the absence of selection for growth on benzene. cleavage sites for the restriction endonucleases dra ... | 1993 | 8500007 |
| creatinase in its collapsed a state shows properties of a molten globule with dimeric quaternary structure. | in the past, the molten globule state at acidic ph (a state) has mainly been observed for small single-domain proteins. for more complex proteins such as immunoglobulin, alternatively folded states, with certain characteristics of the molten globule but different thermodynamic properties, were observed. in the present work, the acid-induced structural characteristics of a homodimer, creatinase from pseudomonas putida, are described. the 91-kda protein at ph 2 shows molten globule behavior in tha ... | 1993 | 8504814 |
| purification of pseudomonas putida acyl coenzyme a ligase active with a range of aliphatic and aromatic substrates. | acyl coenzyme a (acyl-coa) ligase (acyl-coa synthetase [acoas]) from pseudomonas putida u was purified to homogeneity (252-fold) after this bacterium was grown in a chemically defined medium containing octanoic acid as the sole carbon source. the enzyme, which has a mass of 67 kda, showed maximal activity at 40 degrees c in 10 mm k2po4h-napo4h2 buffer (ph 7.0) containing 20% (wt/vol) glycerol. under these conditions, acoas showed hyperbolic behavior against acetate, coa, and atp; the kms calcula ... | 1993 | 8476289 |
| superoxide dismutase activity in root-colonizing pseudomonads. | several saprophytic fluorescent pseudomonads that are aggressive root colonizers express similar specific activities of superoxide dismutase during growth in liquid culture. the pseudomonads have the potential to produce hydrogen peroxide sensitive and hydrogen peroxide insensitive isoforms of superoxide dismutase with distinct mobilities in nondenaturing polyacrylamide gel electrophoresis. synthesis of the hydrogen peroxide insensitive form is enhanced by limited iron availability, by exposure ... | 1993 | 8500011 |
| diketocamphane enantiomer-specific 'baeyer-villiger' monooxygenases from camphor-grown pseudomonas putida atcc 17453. | pseudomonas putida atcc 17453 grew with either (+)- or (-)-camphor as sole carbon source. enantiomer-specific 'biological baeyer-villiger' monooxygenases were synthesized irrespective of the camphor isomer used for growth. the two enzymes are probably the products of separate genes but showed many similarities. each consisted of two electrophoretically identical subunits, bound flavin mononucleotide (fmn) non-covalently and accepted electrons from an induced nadh dehydrogenase which interacted w ... | 1993 | 8515237 |
| physical organization of the upper pathway operon promoter of the pseudomonas tol plasmid. sequence and positional requirements for xylr-dependent activation of transcription. | the upper pathway operon of the pseudomonas putida tol plasmid belongs to the -12/-24 class of promoters. these promoters exhibit three regions critical for regulated transcription, namely, the -12/-24 site for rna polymerase/sigma 54 binding, the -55/-67 region for ihf protein binding, and the -130(uas2)/-170(uas1) region, where two sites for xylr binding are located. the xylr-protected g residues located at -131, -139, -160 and -169 were replaced with as, and the activity of the mutant promote ... | 1993 | 8510657 |
| comparison of the nucleotide sequences of the meta-cleavage pathway genes of tol plasmid pww0 from pseudomonas putida with other meta-cleavage genes suggests that both single and multiple nucleotide substitutions contribute to enzyme evolution. | tol plasmid pww0 from pseudomonas putida mt-2 encodes catabolic enzymes required for the oxidation of toluene and xylenes. the structural genes for these catabolic enzymes are clustered into two operons, the xylcmabn operon, which encodes a set of enzymes required for the transformation of toluene/xylenes to benzoate/toluates, and the xylxyzltegfjqkih operon, which encodes a set of enzymes required for the transformation of benzoate/toluates to krebs cycle intermediates. the latter operon can be ... | 1993 | 8510667 |
| induction and characterization of a cytochrome p-450-dependent camphor hydroxylase in tissue cultures of common sage (salvia officinalis). | (+)-camphor, a major monoterpene of the essential oil of common sage (salvia officinalis), is catabolized in senescent tissue, and the pathway for the breakdown of this bicyclic ketone has been previously elucidated in sage cell-suspension cultures. in the initial step of catabolism, camphor is oxidized to 6-exo-hydroxycamphor, and the corresponding nadph- and o2-dependent hydroxylase activity was demonstrated in microsomal preparations of sage cells. several well-established inhibitors of cytoc ... | 1993 | 12231778 |
| putative functions of phenylalanine-350 of pseudomonas putida cytochrome p-450cam. | cytochrome p-450cam hydroxylates d-camphor, using molecular oxygen and reducing equivalents transferred via putidaredoxin. we constructed mutant genes in which phe-350 of p-450cam was replaced by leu, tyr, or his by site-directed mutagenesis, expressed them in escherichia coli, purified the mutant proteins, and compared their enzymic properties with those of the wild type p-450cam. nadh oxidation rate of the tyr mutant in the reconstituted system with putidaredoxin and putidaredoxin reductase wa ... | 1994 | 8305479 |
| the role of lysine 166 in the mechanism of mandelate racemase from pseudomonas putida: mechanistic and crystallographic evidence for stereospecific alkylation by (r)-alpha-phenylglycidate. | the mechanism of irreversible inactivation of mandelate racemase (mr) from pseudomonas putida by alpha-phenylglycidate (alpha pga) has been investigated stereochemically and crystallographically. the (r) and (s) enantiomers of alpha pga were synthesized in high enantiomeric excess (81% ee and 83% ee, respectively) using sharpless epoxidation chemistry. (r)-alpha pga was determined to be a stereospecific and stoichiometric irreversible inactivator of mr. (s)-alpha pga does not inactivate mr and a ... | 1994 | 8292591 |
| responses to nutrient starvation in pseudomonas putida kt2442: analysis of general cross-protection, cell shape, and macromolecular content. | the physiology of pseudomonas putida kt2442 with respect to growth and carbon starvation was studied. during the transition from growth to nongrowth, the cell shape changes from cylindrical to spheric, a change which is accompanied by reductions in cell size, dna and ribosome content, and the rate of total protein synthesis. in addition, a pattern of general cross-protection develops, which enables the cells to survive environmental stresses such as high and low temperatures, elevated osmolarity ... | 1994 | 8282712 |
| mechanism of p-hydroxyphenylacetate-3-hydroxylase. a two-protein enzyme. | p-hydroxyphenylacetate-3-hydroxylase purified from pseudomonas putida is a two-protein enzyme requiring a flavoprotein and a coupling protein for productive hydroxylation (arunachalam, u., massey, v., and vaidyanathan, c. s. (1992) j. biol. chem. 267, 25848-25855). this paper presents information on the mechanism of the enzyme from absorbance and fluorescence stopped-flow studies. the reduction of the substrate-free flavoprotein by nadh was slow and was not altered by the presence of the couplin ... | 1994 | 8276789 |
| fpta, the fe(iii)-pyochelin receptor of pseudomonas aeruginosa: a phenolate siderophore receptor homologous to hydroxamate siderophore receptors. | the pseudomonas aeruginosa siderophore pyochelin is structurally unique among siderophores and possesses neither hydroxamate- nor catecholate-chelating groups. the structural gene encoding the 75-kda outer membrane fe(iii)-pyochelin receptor fpta has been isolated by plasmid rescue techniques and sequenced. the n-terminal amino acid sequence of the isolated fpta protein corresponded to that deduced from the nucleotide sequence of the fpta structural gene. the mature fpta protein has 682 amino ac ... | 1994 | 8288523 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |