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purification and some properties of a 2fe ferredoxin in pseudomonas ovalis.a [2fe-2s] ferredoxin was found in pseudomonas ovalis which was grown in a medium supplemented with glucose and ammonium sulfate. the molecular weight of the 2fe ferredoxin was estimated to be 13,000. it contained 2.2 gramatoms of non-heme iron and 2.3 gramatoms of acid-labile sulfur per mole protein. the absorption and circular dichroism spectra were characteristic of those of [2fe-2s] type ferredoxins, especially adrenodoxin and putidaredoxin. the electron paramagnetic resonance spectrum of th ...19892548508
molecular cloning, coding nucleotides and the deduced amino acid sequence of p-450bm-1 from bacillus megaterium.the gene encoding barbiturate-inducible cytochrome p-450bm-1 from bacillus megaterium atcc 14581 has been cloned and sequenced. an open reading frame in the 1.9 kb of cloned dna correctly predicted the nh2-terminal sequence of p-450bm-1 previously determined by protein sequencing, and, in toto, predicted a polypeptide of 410 amino acid residues with an mr of 47,439. the sequence is most, but less than 27%, similar to that of p-450cam from pseudomonas putida, so that p-450bm-1 clearly belongs to ...19892597681
[study of morphology and genome structure of pseudomonas putida bacteriophages for their classification].a group of 27 bacteriophages specific for pseudomonas putida strains ppg1 and ppn has been isolated. the phages were characterized and compared with the previously described virulent (pf 16, af, tf and pmw) and temperate (pp56 and pp71) phages. the new phages belong to b1 and c1 morphotypes, according to ackerman's classification. phage dnas were digested with several endonucleases; the molecular weights and homology of the dnas were determined. all phages of p. putida isolated up to now were di ...19892599372
nucleotide and deduced amino acid sequence of the rpon sigma-factor of pseudomonas putida. 19892602128
a simple procedure for transferring genes cloned in escherichia coli vectors into other gram-negative bacteria: phenotypic analysis and mapping of tol plasmid gene xylk.a simple method to transfer non-conjugative escherichia coli plasmids to other gram-negative bacteria and their maintenance is described. this method involves generation of inverse transposition-mediated cointegrates of the non-conjugative e. coli plasmid with a conjugative incw broad-host-range plasmid, r388, carrying tn10. isolation of such cointegrates was readily effected by conjugal transfer from an e. coli donor containing the two plasmids to an e. coli recipient, with selection for transc ...19892548929
characterization of five genes in the upper-pathway operon of tol plasmid pww0 from pseudomonas putida and identification of the gene products.the upper operon of the tol plasmid pww0 of pseudomonas putida encodes a set of enzymes which transform toluene and xylenes to benzoate and toluates. the genetic organization of the operon was characterized by cloning of the upper operon genes into an expression vector and identification of their products in escherichia coli maxicells. this analysis showed that the upper operon contains at least five genes in the order of xylc-xylm-xyla-xylb-xyln. between the promoter of the operon and xylc, the ...19892549010
cloning of pmol28-encoded nickel resistance genes and expression of the genes in alcaligenes eutrophus and pseudomonas spp.the 163-kilobase-pair (kb) plasmid pmol28, which determines inducible resistance to nickel, cobalt, chromate, and mercury salts in its native host alcaligenes eutrophus ch34, was transferred to a derivative of a. eutrophus h16 and subjected to cloning procedures. after tn5 transposon mutagenesis, restriction endonuclease analysis, and dna-dna hybridization, two dna fragments, a 9.5-kb kpni fragment and a 13.5-kb hindiii fragment (hki), were isolated. hki contained ek1, the kpni fragment, as a su ...19892549012
[plasmids for biphenyl, chlorobiphenyl and metatoluylate degradation from pseudomonas putida].pseudomonas putida strain su83, harbors the pbs311 plasmid coding for the degradation of biphenyl, 2- and 4-chlorbiphenyl, meta- and paratoluylate. the insertional mutants of the plasmid obtained by the transposon tn5 insertion were isolated. one of the mutants was used for cloning of the biphenyl degradation genes. the plasmid pbs311:: tn5 dna was inserted into the bamhi site of the plasmid pbr322 and cloned. 11 recombinants of 354 tested were treated with 0.1% solution of 2,3-dioxybiphenyl. on ...19892628753
[genetic determination of degradation of ampholytic surfactants].plasmid dna was detected in pseudomonas putida 141 and p. stutzeri at strains which caused destruction of the ampholytic surfactants alkylamino-bis-propionate (aabp) and amidobetaine, respectively. as was demonstrated using genetic analytic procedures, the plasmids controlled aabp and amidobetaine destruction. no plasmid dna was found in p. desmolytica c37 which caused cyclimide destruction or in pseudomonas sp. 1 and citrobacter freundii to strains responsible for aabp destruction. apparently, ...19892636974
extracellular product of nocardia amarae induces bacterial cell flocculation.the fact that nocardia amarae yk1 produced a bacterial flocculation-inducing substance (designated as fix) was discovered. fix had a function of flocculating proliferous cells. fix-induced flocculation was inhibited by making cells resting, but not completely by adding chloramphenicol. fix worked widely on gram-positive to -negative bacteria. in the presence of fix, achromobacter cycloclastus iam1013, acinetobacter calcoaceticus iam1517, bacillus subtilis iam1069, escherichia coli c600-1, e. col ...19892653957
the purification and characterization of 4-ethylphenol methylenehydroxylase, a flavocytochrome from pseudomonas putida jd1.the enzyme 4-ethylphenol methylenehydroxylase was purified from pseudomonas putida jd1 grown on 4-ethylphenol. it is a flavocytochrome c for which the mr was found to be 120,000 by ultracentrifuging and 126,000 by gel filtration. the enzyme consists of two flavoprotein subunits each of mr 50,000 and two cytochrome c subunits each of mr 10,000. the redox potential of the cytochrome is 240 mv. hydroxylation proceeds by dehydrogenation and hydration to give 1-(4'-hydroxyphenyl)ethanol, which is als ...19892556994
transposable elements for efficient manipulation of a wide range of gram-negative bacteria: promoter probes and vectors for foreign genes.we describe here the construction and use of a series of modified transposons based on the insertion sequence is1. like their parent, omegon-km [fellay et al., gene 76 (1989) 215-226], these elements permit efficient insertional mutagenesis of a variety of gram-negative bacteria. the presence of a functional pbr322 origin of replication within the transposable element facilitates subsequent cloning of the mutated gene. the omegon-km system was previously shown to function in pseudomonas putida, ...19892559879
[cloning and gene expression determining phenol breakdown in pseudomonas putida strains]. 19892561419
[cloning of pseudomonas putida genes responsible for the primary stages of oxidation of naphthalene in escherichia coli cells].data on cloning pseudomonas putida d-plasmid pbs286 (incp-9) genes which are responsible for primary stages of naphthalene oxidation as well as data on the expression of these genes in escherichia coli cells are presented. recombinant plasmid pbs959 containing the whole constitutive naha locus encoding naphthalene dioxygenase, a key enzyme of the naphthalene oxidation pathway, has been constructed on the basis of the puc19 vector. an evidence has been obtained that at least a portion of the sequ ...19892661326
plasmid control of the pseudomonas aeruginosa and pseudomonas putida phenotypes and of linalool and p-cymene oxidation.two pseudomonas strains (ppg777 and pag158) were derived from the parent isolate pseudomonas incognita (putida). strain ppg777 resembles the parental culture in growth on linalool as a source of carbon and slight growth on p-cymene, whereas pag158 grows well on p-cymene, but not on linalool or other terpenes tested, and has a p. aeruginosa phenotype. curing studies indicate that linalool metabolism is controlled by an extrachromosomal element whose loss forms a stable strain pag158 with the p-cy ...19892504698
selection and characterization of a mutant of the cloned gene for mandelate racemase that confers resistance to an affinity label by greatly enhanced production of enzyme.the plasmid pscr1 containing the gene for mandelate racemase (ec 5.1.2.2) from pseudomonas putida (atcc 12633) allows pseudomonas aeruginosa (atcc 15692) to grow on (r)-mandelate as its sole carbon source [ransom, s. c., gerlt, j. a., powers, v. m., & kenyon, g. l. (1988) biochemistry 27, 540]; the chromosome of the p. aeruginosa host apparently does not contain the gene for mandelate racemase but does contain genes for the remaining enzymes in the mandelate pathway and enables growth on (s)-man ...19892496759
oxygen diffusivity in gel beads containing viable cells.this article proposes a simple steady-state method for measuring the effective diffusion coefficient of oxygen (d(e)) in gel beads entrapping viable cells. we applied this method to the measurement of d(e) in ca- and ba-alginate gel beads entrapping saccharomyces cerevisiae and pseudomonas ovalis. the diffusivity of oxygen through gel beads containing viable cells was measured within an accuracy of +/-7% and found not to be influenced by cell density (0-30 g/l gel), cell type, and cell viability ...198918588184
autoradiographic determination of mass-transfer limitations in immobilized cell reactors.pseudomonas putida cells were grown in confined volumes in dual-membrane immobilized cell reactors constructed from microporous polyethylene hollow fibers and silicone rubber tubules as a model system for the study of mass transport in microbial aggregates. local cell concentrations in the reactors reached 300 g dry mass/l. pulse-chase radioisotope labeling with (35)so(4) (2-) was used to estimate the rates of cell mass synthesis and degradation. sulfur incorporation consistently exceeded sulfur ...198918588110
genes for two herbicide-inducible cytochromes p-450 from streptomyces griseolus.streptomyces griseolus atcc 11796 contains two inducible, herbicide-metabolizing cytochromes p-450 previously designated p-450su1 and p-450su2 (p-450cva1 and p-450cvb1, respectively, using nomenclature of nebert et al. [d. w. nebert, m. adesnik, m. j. coon, r. w. estabrook, f. j. gonzalez, f. p. guengerich, i. c. gunsalus, e. f. johnson, b. kemper, w. levin, i. r. phillips, r. sato, and m. r. waterman, dna 6:1-11, 1987]). using antibodies directed against cytochrome p-450su1, its n-terminal amin ...19902345149
protein components of a cytochrome p-450 linalool 8-methyl hydroxylase.the cytochrome p-450 heme-thiolate monooxygenases that hydroxylate monoterpene hydrocarbon groups are effective models for the cytochrome p-450 family. we have purified and characterized the three proteins from a p-450-dependent linalool 8-methyl hydroxylase in pseudomonas putida (incognita) strain ppg777. the proteins resemble the camphor 5-exohydroxylase components in chemical and physical properties; however, they show neither immunological cross-reactivity nor catalytic activity in heterogen ...19902295633
biotransformation of substituted benzoates to the corresponding cis-diols by an engineered strain of pseudomonas oleovorans producing the tol plasmid-specified enzyme toluate-1,2-dioxygenase.the conversion of substituted benzoates into 1,2-cis-dihydroxycyclohexa-3,5-diene carboxylic acids (cis-diols) was effected by using escherichia coli and pseudomonas recombinants carrying the xylxyz genes originating from the pseudomonas putida mt-2 tol plasmid, thus producing toluate-1,2-dioxygenase. pseudomonas oleovorans gpo12 recombinants readily produced meta- and para-substituted cis-diols, but were limited in their oxidation of ortho-substituted substrates.19902306096
the meta cleavage operon of tol degradative plasmid pww0 comprises 13 genes.the meta-cleavage operon of tol plasmid pww0 of pseudomonas putida encodes a set of enzymes which transform benzoate/toluates to krebs cycle intermediates via extradiol (meta-) cleavage of (methyl)catechol. the genetic organization of the operon was characterized by cloning of the meta-cleavage genes into an expression vector and identification of their products in escherichia coli maxicells. this analysis showed that the meta-cleavage operon contains 13 genes whose order and products (in kiloda ...19902183008
purification and characterization of d-2-haloacid dehalogenase from pseudomonas putida strain aj1/23.a d-2-haloacid dehalogenase was isolated and purified to homogeneity from pseudomonas putida strain aj1/23. the enzyme catalysed the stereospecific dehalogenation of the d-isomer of 2-chloropropionate. using a new ion-chromatograph assay, the enzyme was found to catalyse the dehalogenation of short-chain 2-halocarboxylic acids. maximum enzyme activity occurred at ph 9.5 and 50 degrees c and the enzyme was insensitive to most -sh reagents. the enzyme has an mr of about 135,000 and appears to be c ...19902380688
growth-phase-dependent expression of the pseudomonas putida tol plasmid pww0 catabolic genes.pseudomonas putida tol plasmid pww0 catabolic genes are clustered into two operons. the first, the upper operon, is controlled by the xylr regulatory gene, whereas the second, the meta operon, is controlled by the xyls regulatory gene. the xyls gene itself is subjected to control by xylr. in this study, we show that the tol catabolic operons were poorly induced in cells growing at the early-exponential-growth phase but strongly induced in cells at late-exponential-growth phase. we constructed fu ...19902254244
degradation of 3-chlorobiphenyl by in vivo constructed hybrid pseudomonads.3-chlorobiphenyl-degrading bacteria were obtained from the mating between pseudomonas putida strain bn10 and pseudomonas sp. strain b13. strains such as bn210 resulted from the transfer of the genes coding the enzyme sequence for the degradation of chlorocatechols from b13 into bn10, whereas b13 derivatives such as b131 have acquired the biphenyl degradation sequence from bn10. during growth of the hybrid strains on 3-chlorobiphenyl 90% chloride was released. activities of phenylcatechol 2,3-dio ...19902276606
three isozymes of catechol 1,2-dioxygenase (pyrocatechase), alpha alpha, alpha beta, and beta beta, from pseudomonas arvilla c-1.three isozymes of catechol 1,2-dioxygenase (pyrocatechase) from pseudomonas arvilla c-1 were separated using deae-toyopearl chromatography. the specific activities of each isozyme were similar to one another. the molecular weights of isozymes 1, 2, and 3 were estimated to be approximately 67,000, 64,000, and 59,000, respectively, from gel filtration. on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isozymes 1 and 3 gave a single protein band, corresponding to mr = 32,000 and 30,000, ...19902295613
isolation and partial characterization of an extradiol non-haem iron dioxygenase which preferentially cleaves 3-methylcatechol.a purification procedure has been developed for an extradiol dioxygenase expressed in escherichia coli, which was originally derived from a pseudomonas putida strain able to grow on toluidine. physical and kinetic properties of the enzyme have been investigated. the enzyme has a subunit mr of 33,500 +/- 2000 by sds/polyacrylamide-gel electrophoresis. gel filtration indicates a molecular mass under non-denaturing conditions of 120,000 +/- 20,000. the n-terminal sequence (35 residues) of the enzym ...19902317207
chemotaxis of pseudomonas putida toward chlorinated benzoates.the chlorinated aromatic acids 3-chlorobenzoate and 4-chlorobenzoate are chemoattractants for pseudomonas putida prs2000. these compounds are detected by a chromosomally encoded chemotactic response to benzoate which is inducible by beta-ketoadipate, an intermediate of benzoate catabolism. plasmid pac27, encoding enzymes for 3-chlorobenzoate degradation, does not appear to carry genes for chemotaxis toward chlorinated compounds.19902339899
functional modification of an arginine residue on salicylate hydroxylase.salicylate hydroxylase from pseudomonas putida (ec 1.14.13.1, salicylate, nadh:oxygen oxidoreductase) is an fad-containing monooxygenase, which catalyzes decarboxylative hydroxylation of salicylate to produce catechol in the presence of nadh and o2. by chemical treatment of the enzyme with dicarbonyl reagents, such as glyoxal, the original oxygenase activity was converted to the salicylate-dependent nadh-dehydrogenase activity with free fad as electron acceptor. one of twenty arginine residues o ...19902223838
[new plasmids of herbicide 2,4-dichlorophenoxyacetic acid biodegradation].three herbicide 2,4-d metabolizing bacterial strains were isolated from three independent soil samples of estonia. the strains, although belonging to various species, contain 2,4-d degradative plasmids with identical restriction patterns. pest4001 is a 78 kb conjugative plasmid. all pseudomonas putida paw340 2,4-d+ transconjugants obtained a 70 kb plasmid pest4011 - a deletion derivative of the pest4001. the restriction patterns of the plasmids mentioned above are considerably different from tho ...19902373362
cloning and sequence analysis of the genes encoding the alpha and beta subunits of the e1 component of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus.a 4175-bp ecori fragment of dna that encodes the alpha and beta chains of the pyruvate dehydrogenase (lipoamide) component (e1) of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus has been cloned in escherichia coli. its nucleotide sequence was determined. open reading frames (pdha, pdhb) corresponding to the e1 alpha subunit (368 amino acids, mr 41,312, without the initiating methionine residue) and e1 beta subunit (324 amino acids, mr 35,306, without the initiating ...19902200674
complete amino acid sequence of the cytochrome subunit and amino-terminal sequence of the flavin subunit of flavocytochrome c (sulfide dehydrogenase) from chlorobium thiosulfatophilum.the complete amino acid sequence of the 86-residue heme subunit of flavocytochrome c (sulfide dehydrogenase) from the green phototrophic bacterium chlorobium thiosulfatophilum strain tassajara has been determined as follows: apeqsksiprgeilslscagchgtdgksesiiptiygrsaeyiesalldfksga- rpstvmgrhakgysdeeihqiaeyfgslstmnn. the subunit has a single heme-binding site near the n terminus, consisting of a pair of cysteine residues at positions 18 and 21. the out-of-plane ligands are apparently contributed by ...19902161842
[studies on respiratory infections in primary care clinic (iii). distribution of bacteria isolated from patients with respiratory infections visiting 21 private clinics in the tohoku district of japan].the bacteriology of the isolates from the throat swab and the sputum respectively of 2,539 patients with respiratory infections visiting 21 private clinics in tohoku district of japan during the period from january to april in 1989 was documented. of the 2,539 patients, 1,694 had an acute upper respiratory infection, 609 had acute bronchitis, 46 had acute pneumonia, 84 had acute exacerbation of chronic respiratory infections and 106 had respiratory infections without diagnosis registered. 1887 ( ...19902243193
purification and characterization of s-alkylcysteine alpha,beta-lyase from pseudomonas putida.s-alkylcysteine alpha,beta-lyase [ec 4.4.1.6] was purified to more than 90% homogeneity from the cell extract of pseudomonas putida icr 3640. the enzyme has a molecular weight of about 195,000, and is composed of six subunits identical in molecular weight (37,000). pyridoxal 5'-phosphate is required as a cofactor. the enzyme catalyzes the alpha,beta-elimination of s-methyl-l-cysteine and its analogs such as s-ethyl-l-cysteine, l-djenkolate, se-methyl-dl-selenocysteine, and o-methyl-l-serine. how ...19902081976
[the preparation and properties of catechol-1,2-dioxygenase from pseudomonas putida].catechol-1,2-dioxygenase (ec 1.13.11.1) catalyzes the degradation of catechol to cis, cis-muconic acid. the biochemical properties of catechol-1,2-dioxygenase from pseudomonas putida 84103 were investigated. the optimum ph and temperature is 7.5-8.0 and 25-30 degrees c, respectively. cu2+, zn2+ inhibit the enzyme activity. the paper chromatograph and uv absorption spectrum of enzymatic reaction product are accordance with those of the standard muconic acid.19902251833
xanthine dehydrogenase and 2-furoyl-coenzyme a dehydrogenase from pseudomonas putida fu1: two molybdenum-containing dehydrogenases of novel structural composition.the constitutive xanthine dehydrogenase and the inducible 2-furoyl-coenzyme a (coa) dehydrogenase could be labeled with [185w]tungstate. this labeling was used as a reporter to purify both labile proteins. the radioactivity cochromatographed predominantly with the residual enzymatic activity of both enzymes during the first purification steps. both radioactive proteins were separated and purified to homogeneity. antibodies raised against the larger protein also exhibited cross-reactivity toward ...19902170335
selection of independent plasmids determining phenol degradation in pseudomonas putida and the cloning and expression of genes encoding phenol monooxygenase and catechol 1,2-dioxygenase.long-term cultivation of the pseudomonas putida multiplasmid strain est1020 on phenol resulted in the formation of individual phe plasmids determining phenol degradation. four types of phe plasmids, pest1024, pest1026, pest1028, and pest1029, are characterized. they all contain a transferrable replicon similar to pwwo-8 with a partly duplicated dna sequence of the 17-kb transposable element of this plasmid and include various amounts of dna that carry genes encoding phenol degradation (phe genes ...19902270227
cloning and expression of the plasmid-encoded benzene dioxygenase genes from pseudomonas putida ml2.hybridization using heterologous dioxygenase gene probes indicated the presence of the genes encoding the enzyme benzene dioxygenase on a 112 kb plasmid from pseudomonas putida ml2. they were identified as benzene dioxygenase genes (bed abc1c2) by cloning in escherichia coli and analysis of expression by western blotting using antibodies raised to the four polypeptides of purified benzene dioxygenase.19902083838
genetic analysis of chromosomal operons involved in degradation of aromatic hydrocarbons in pseudomonas putida tmb.the catabolic pathway for the degradation of aromatic hydrocarbons encoded by pseudomonas putida tmb differs from the tol plasmid-encoded pathway as far as regulation of the upper pathway is concerned. we found, by analyzing tn5-induced mutants and by southern blot hybridization with appropriate probes derived from the tol plasmid pww0, that the catabolic genes of strain tmb were located on the bacterial chromosome and not on the 84-kb plasmid harbored by this strain. the catabolic genes of tmb ...19902172213
transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria.a simple procedure for cloning and stable insertion of foreign genes into the chromosomes of gram-negative eubacteria was developed by combining in two sets of plasmids (i) the transposition features of tn10 and tn5; (ii) the resistances to the herbicide bialaphos, to mercuric salts and organomercurial compounds, and to arsenite, and (iii) the suicide delivery properties of the r6k-based plasmid pgp704. the resulting constructions contained unique noti or sfii sites internal to either the tn10 o ...19902172216
cis-1,2-dihydroxycyclohexa-3,5-diene (nad) oxidoreductase (cis-benzene dihydrodiol dehydrogenase) from pseudomonas putida ncib 12190. 19902280699
toluene dioxygenase from pseudomonas putida f1. 19902280710
benzene dioxygenase from pseudomonas putida, ml2 (ncib 12190). 19902280718
[cloning genes for biosynthesis of pseudomonas putida tryptophan in escherichia coli cells].the trpe, trpc and trpiba genes of pseudomonas putida were cloned by complementation of the corresponding auxotrophic mutations of escherichia coli using pbr322 as a vector. with the exception of trpe, transcription of all genes in new host takes place under control of their own promoters. expression of the trpd gene linked to trpc was not registered in e. coli. repressible trpc enzyme was synthesized constitutively in e. coli. characteristic regulation of p. putida trpba genes via induction by ...19902283047
comparative in vitro activities of newer quinolones against pseudomonas species and xanthomonas maltophilia isolated from patients with cancer.the in vitro susceptibilities of three pseudomonas species (pseudomonas aeruginosa, pseudomonas putida, and pseudomonas fluorescens) and xanthomonas maltophilia to quinolone antimicrobial agents were determined. several newer agents, particularly pd117558, pd117596, pd127391, sparfloxacin (at-4140), a-56620, and temafloxacin, were active against pseudomonas species. x. maltophilia isolates were generally less susceptible than were pseudomonas isolates but were inhibited by some of the newer quin ...19902285297
[rare initiation codons are regulators of expression of the rpoc gene].translation of the rpoc genes in escherichia coli and salmonella typhimurium is known to start from the gug codon. now, using toeprint analysis we have shown uug to be the initiation codon of the pseudomonas putida rpoc gene. if3 does not seem to proofread initiation at the uug codon. the rpoc genes of p. putida, e. coli, and s. typhimurium, which use rare start codons, have strong sd-domains aggagg (p. p.) and gggag (e. c., s. t.), optimal seven-nucleotide spacing between sd and start codons, a ...19902285427
pseudomonas putida kf715 bphabcd operon encoding biphenyl and polychlorinated biphenyl degradation: cloning, analysis, and expression in soil bacteria.we cloned the entire bphabcd genes encoding degradation of biphenyl and polychlorinated biphenyls to benzoate and chlorobenzoates from the chromosomal dna of pseudomonas putida kf715. the nucleotide sequence revealed two open reading frames corresponding to the bphc gene encoding 2,3-dihydroxybiphenyl dioxygenase and the bphd gene encoding 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (ring-meta-cleavage compound) hydrolase.19902105297
putidaredoxin reductase and putidaredoxin. cloning, sequence determination, and heterologous expression of the proteins.the oxidation of camphor by cytochrome p-450cam requires the participation of a flavoprotein, putidaredoxin reductase, and an iron-sulfur protein, putidaredoxin, to mediate the transfer of electrons from nadh to p-450 for oxygen activation. a 2.2-kilobase pair bamhi-stui fragment from whole cell dna of camphor-grown pseudomonas putida has been cloned and sequenced. translation of the sequence revealed two open reading frames that could code for putidaredoxin reductase and putidaredoxin. in the c ...19902180940
dna sequences of genes encoding acinetobacter calcoaceticus protocatechuate 3,4-dioxygenase: evidence indicating shuffling of genes and of dna sequences within genes during their evolutionary divergence.the dna sequence of a 2,391-base-pair hindiii restriction fragment of acinetobacter calcoaceticus dna containing the pcachg genes is reported. the dna sequence reveals that a. calcoaceticus pca genes, encoding enzymes required for protocatechuate metabolism, are arranged in a single transcriptional unit, pcaefdbchg, whereas homologous genes are arranged differently in pseudomonas putida. the pcag and pcah genes represent separate reading frames respectively encoding the alpha and beta subunits o ...19902298704
a new family of rsf1010-derived expression and lac-fusion broad-host-range vectors for gram-negative bacteria.a series of broad-host-range expression and lac fusion vectors, based on rsf1010 derivatives, was constructed. the expression vectors contain various promoters (pnm, plac, ptac and ps1) for expression of foreign genes. the efficiency of the promoters was determined in escherichia coli, rhizobium meliloti, rhizobium leguminosarum and pseudomonas putida by beta-galactosidase activity measurements. of the promoters assayed in e. coli, the most effective is the tac promoter, whereas in soil bacteria ...19902115488
cloning and expression of genes involved in 4-chlorobiphenyl transformation by pseudomonas testosteroni: homology to polychlorobiphenyl-degrading genes in other bacteria.the genes of pseudomonas testosteroni strain b-356, specifying the transformation of 4-chlorobiphenyl (4-cb) into 4-chlorobenzoic acid (4-cba) were cloned into pseudomonas putida kt2440 using a broad-host-range cosmid, ppsa842. of 10,000 clones tested, four were able to transform 4-cb. gas chromatographic and mass spectrometric analysis of the catabolic products from two of the 4-cb-transforming clones carrying the hybrid plasmids, pda1 and pda2, demonstrated that pda1 carried a complete set of ...19902311936
streptomyces promoter-probe plasmids that utilise the xyle gene of pseudomonas putida. 19902315034
a family of positive regulators related to the pseudomonas putida tol plasmid xyls and the escherichia coli arac activators.the xyls family consists of a least 8 different transcriptional regulators. six of these proteins are positive regulators for the catabolism of carbon sources (benzoate and sugars) in escherichia coli, pseudomonas putida and erwinia carotovora, and two of them are involved in pathogenesis in escherichia coli and yersinia enterocolitica. based on protein alignments, the members of this family exhibit a long stretch of homology at the c-terminal end. the regulators involved in the catabolism of ca ...19902186376
nucleotide sequence and expression of the isoamylase gene from an isoamylase-hyperproducing mutant, pseudomonas amyloderamosa jd210.the isoamylase gene (iso) of pseudomonas amyloderamosa jd210, an isoamylase-hyperproducing mutant, was cloned in an isoamylase-deficient and transformable mutant strain k31. by deletion analysis, the iso gene was found to be located within a 3.3 kilobases bamhi fragment. its nucleotide sequence contained an open reading frame of 2328 nucleotides (776 amino acids) encoding a secreted isoamylase precursor. the iso gene fragment was inserted into plasmids pkt230 and pbr 322 in opposite orientations ...19902248978
isolation of high frequency of recombination donors from tn5 chromosomal mutants of pseudomonas putida ppn and recalibration of the genetic map.a tn5 loaded derivative of the incp-10 plasmid r91-5 (pmo75) was used as a suicide vector to generate random chromosomal insertion mutations in pseudomonas putida ppn. reintroduction of pmo75 into such mutants resulted in integration of the plasmid at the site of tn5 insertion, giving rise to two classes of high frequency of donors recombination (hfr) donors, transferring chromosome at high frequency (greater than 10(-1) per donor cell) in opposite directions. consequently, tn5 induced auxotroph ...19902174392
sequence analysis of the huth gene encoding histidine ammonia-lyase in pseudomonas putida.the complete nucleotide sequence of the huth gene, encoding histidine ammonia-lyase (histidase), in pseudomonas putida atcc 12633 has been determined from the appropriate portions of the hut region that had been cloned into escherichia coli. the resulting dna sequence revealed an open reading frame of 1,530 base pairs, corresponding to a protein subunit of approximate molecular weight 53,600, in the location previously identified for the histidase gene by tn1000 mutagenesis. translation began at ...19902332400
upstream regulatory sequence for transcriptional activator xylr in the first operon of xylene metabolism on the tol plasmid.transcription of the first operon coding for m-xylene-degrading enzymes on the tol plasmid of pseudomonas putida is activated by the xylr gene product in the presence of m-xylene. the operon has the consensus sequence of the ntr/nif promoters at -24 and -12 regions, and the transcription is dependent on an rna polymerase containing a sigma factor ntra (rpon or sigma 54). deletion derivatives of the upstream sequence of the operon promoter were made in vitro and connected with the xyle gene on a ...19902174974
characterization of the multiple catalytic activities of tartrate dehydrogenase.tartrate dehydrogenase (tdh) has been purified to apparent homogeneity from pseudomonas putida and has been demonstrated to catalyze three different nad(+)-dependent reactions. tdh catalyzes the oxidation of (+)-tartrate to form oxaloglycolate and the oxidative decarboxylation of d-malate to form pyruvate and co2. d-glycerate and co2 are formed from meso-tartrate in a reaction that is formally a decarboxylation with no net oxidation or reduction. the steady-state kinetics of the first two reacti ...19902184888
purification and characterization of salicylate hydroxylase from pseudomonas putida ppg7.the salicylate hydroxylase from p. putida ppg7 was purified and characterized. the enzyme appears to be monomeric, and it showed one major band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with an apparent mr of 45 kda. the sequence of the first 25 amino acids of salicylate hydroxylase (ppg7) was determined. also, the total amino acid composition of salicylate hydroxylase (ppg7) was obtained and compared with that of the known salicylate hydroxylase from p. putida.19902363715
[the sos-like reactions of pseudomonas putida].under ultraviolet radiation of pseudomonas putida 1087 the sos-similar response which is expressed in the inhibition of cell respiration and cell division with the following filamentation is revealed. in the result of introduction of ppe24 and pmh21 plasmids into the cells of p. putida 1087 for inhibition of reca-similar protein the sos-similar response disappears and the basic cell mass dies.19902372559
carbon catabolite regulation of phenylacetyl-coa ligase from pseudomonas putida.phenylacetyl-coa ligase (pa-coa ligase) from p. putida u is a newly described enzyme involved in the aerobic catabolism of phenylacetic acid. the enzyme was specifically induced when p. putida was grown in a chemically defined medium containing phenylacetic acid as the sole carbon source. the induction of pa-coa ligase was delayed by adding easily metabolizable carbon sources to the medium; the effect was more drastic in the presence of glucose. glucose did not cause catabolic inactivation but r ...19902322284
the downfield resonances in the 1h nmr spectra of azotobacter vinelandii and pseudomonas putida seven-iron ferredoxins.pseudomonas putida and azotobacter vinelandii ferredoxins each contain one [4fe-4s] cluster and one [3fe-4s] cluster. their polypeptide chains are nearly identical, differing by only 15 residues out of a total of 106. t1 measurements and temperature dependence studies of the 1h nmr spectrum of each ferredoxin demonstrate that all six resolved downfield resonances are near an iron-sulfur center. the five most downfield resonances are shown to arise from protons on cysteinyl beta-carbons by incorp ...19902373698
[peripheral metabolism of pseudomonal putida transconjugants degrading chloro- and methylaromatic compounds].peripheral metabolism was studied in the pseudomonas putida 37cc transconjugant. in the strain grown on benzoate, pyrocatechase (pc) i with a low activity to chlorocatechols was induced, whereas pcii actively decomposing chlorocatechols was induced during its growth on 3-chlorobenzoic acid. the p. putida 37cc transconjugant grown on alpha-methylstyrene (ms) exerted the activity of both metapyrocatechase (mpc) and pc, whereas in the parent strain p. putida r-1 only mpc was involved in the degrada ...19902374510
electroporation and expression of plasmid pbr322 in klebsiella aerogenes nctc 418 and plasmid prk2501 in pseudomonas putida cym 318.klebsiella aerogenes nctc 418 and pseudomonas putida cym 318 were transformed via high-voltage electroporation with plasmids pbr322 and prk2501, respectively. the number of transformants obtained was dependent on the applied voltage, capacitance, and cell recovery procedure. for example, 7.87 x 10(4) transformants/micrograms dna were obtained at 2500 v, 25 muf when k. aerogenes cells were electroporated with pbr322 dna. a lower voltage (1500) and capacitance (3 muf) yielded 2.4 x 10(3) transform ...19902187074
chromium reduction in pseudomonas putida.reduction of hexavalent chromium (chromate) to less-toxic trivalent chromium was studied by using cell suspensions and cell-free supernatant fluids from pseudomonas putida prs2000. chromate reductase activity was associated with soluble protein and not with the membrane fraction. the crude enzyme activity was heat labile and showed a km of 40 microm cro4(2-). neither sulfate nor nitrate affected chromate reduction either in vitro or with intact cells.19902389940
evolutionary differences in chromosomal locations of four early genes of the tryptophan pathway in fluorescent pseudomonads: dna sequences and characterization of pseudomonas putida trpe and trpgdc.pseudomonas putida possesses seven structural genes for enzymes of the tryptophan pathway. all but one, trpg, which encodes the small (beta) subunit of anthranilate synthase, have been mapped on the circular chromosome. this report describes the cloning and sequencing of p. putida trpe, trpg, trpd, and trpc. in p. putida and pseudomonas aeruginosa, dna sequence analysis as well as growth and enzyme assays of insertionally inactivated strains indicated that trpg is the first gene in a three-gene ...19902404959
microcosm for assessing survival of genetically engineered microorganisms in aquatic environments.laboratory-contained microcosms are important for studying the fate and survival of genetically engineered microorganisms. in this study, we describe a simple aquatic microcosm that utilizes survival chambers in a flowthrough or static renewal system. the model was used to study the survival of genetically engineered and wild-type strains of escherichia coli and pseudomonas putida in the lake water environment. temperature-dependent studies indicated that the genetically engineered microorganism ...19902187407
cloning and expression of the ponb gene, encoding penicillin-binding protein 1b of escherichia coli, in heterologous systems.a fragment from the ponb region of the escherichia coli chromosome comprising the promoterless sequence encoding penicillin-binding protein 1b (pbp 1b) has been cloned in a broad-host-range expression vector under the control of the kanamycin resistance gene promoter present in the vector. the hybrid plasmid (pjp3) was used to transform appropriate strains of salmonella typhimurium, pseudomonas putida, and pseudomonas aeruginosa. in all instances, the coding sequence was expressed in the heterol ...19902198260
purification and characterisation of tol plasmid-encoded benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase of pseudomonas putida.benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase, two enzymes of the xylene degradative pathway encoded by the plasmid tol of a gram-negative bacterium pseudomonas putida, were purified and characterized. benzyl alcohol dehydrogenase catalyses the oxidation of benzyl alcohol to benzaldehyde with the concomitant reduction of nad+; the reaction is reversible. benzaldehyde dehydrogenase catalyses the oxidation of benzaldehyde to benzoic acid with the concomitant reduction of nad+; the re ...19902202600
nucleotide sequence of the gene encoding the repressor for the histidine utilization genes of pseudomonas putida.the hutc gene of pseudomonas putida encodes a repressor which, in combination with the inducer urocanate, regulates expression of the five structural genes necessary for conversion of histidine to glutamate, ammonia, and formate. the nucleotide sequence of the hutc region was determined and found to contain two open reading frames which overlapped by one nucleotide. the first open reading frame (orf1) appeared to encode a 27,648-dalton protein of 248 amino acids whose sequence strongly resembled ...19902203753
nucleotide sequence of the gene encoding the repressor for the histidine utilization genes of klebsiella aerogenes.the hutc gene of klebsiella aerogenes encodes a repressor that regulates expression of the histidine utilization (hut) operons. the dna sequence of a region known to contain hutc was determined and shown to contain two long rightward-reading open reading frames (orfs). one of these orfs was identified as the 3' portion of the hutg gene. the other orf was the hutc gene. the repressor predicted from the hutc sequence contained a helix-turn-helix motif strongly similar to that seen in other dna-bin ...19902203754
transcriptional analysis of the promoter region of the pseudomonas putida branched-chain keto acid dehydrogenase operon.branched-chain keto acid dehydrogenase is a multienzyme complex produced by pseudomonas putida when it is grown in a minimal medium containing branched-chain amino acids. a 1.87-kilobase (kb) dna fragment was cloned and sequenced which contained 0.24 kb of the e1 alpha structural gene and 1.6 kb of upstream dna. there were 854 base pairs (bp) of noncoding dna upstream of bkda1, the first gene of the bkd operon, and 592 bp between the transcriptional and translational starts. the g + c content of ...19902211503
different types of formaldehyde-oxidizing dehydrogenases in nocardia species 239: purification and characterization of an nad-dependent aldehyde dehydrogenase.three different dehydrogenases able to oxidize formaldehyde were found in the gram-positive methylotroph, nocardia sp. 239: an nad-dependent aldehyde dehydrogenase (na-adh), and nad- and factor-dependent formaldehyde dehydrogenase (fd-fdh), and a dye-linked aldehyde dehydrogenase (dl-adh). the ratio of the activities observed for the two nad-linked enzymes varied with growth conditions: batch-wise grown cells had nearly the same activities for both enzymes; in fed batch-wise grown cells (methano ...19902241149
the 2.1-a resolution structure of iron superoxide dismutase from pseudomonas ovalis.the 2.1-a resolution crystal structure of native uncomplexed iron superoxide dismutase (ec 1.15.1.1) from pseudomonas ovalis was solved and refined to a final r factor of 24%. the dimeric structure contains one catalytic iron center per monomer with an asymmetric trigonal-bipyramidal coordination of protein ligands to the metal. each monomer contains two domains, with the trigonal ligands (histidines 74 and 160; aspartate 156) contributed by the large domain and stabilized by an extended hydroge ...19902271564
mandelate pathway of pseudomonas putida: sequence relationships involving mandelate racemase, (s)-mandelate dehydrogenase, and benzoylformate decarboxylase and expression of benzoylformate decarboxylase in escherichia coli.the genes that encode the five known enzymes of the mandelate pathway of pseudomonas putida (atcc 12633), mandelate racemase (mdla), (s)-mandelate dehydrogenase (mdlb), benzoylformate decarboxylase (mdlc), nad(+)-dependent benzaldehyde dehydrogenase (mdld), and nadp(+)-dependent benzaldehyde dehydrogenase (mdle), have been cloned. the genes for (s)-mandelate dehydrogenase and benzoylformate decarboxylase have been sequenced; these genes and that for mandelate racemase [ransom, s. c., gerlt, j. a ...19902271624
purification and biochemical characterization of phenylacetyl-coa ligase from pseudomonas putida. a specific enzyme for the catabolism of phenylacetic acid.a new enzyme, phenylacetyl-coa ligase (amp-forming) (pa-coa ligase, ec 6.2.1-) involved in the catabolism of phenylacetic acid (paa) in pseudomonas putida is described and characterized. pa-coa ligase was specifically induced by paa when p. putida was grown in a chemically defined medium in which phenylacetic acid was the sole carbon source. hydroxyl, methyl-phenylacetyl derivatives, and other paa close structural molecules did not induce the synthesis of this enzyme and neither did acetic, buty ...19902324116
molecular cloning of the xyll-xyle region from the p. putida tol plasmid, pdk1.a 5.2 kilobase ecori restriction fragment from the pseudomonas putida hs1 tol plasmid pdk1, encoding a portion of the lower toluene degradation pathway, was cloned into the e. coli plasmid pbr325. a detailed map of the restriction endonuclease sites was constructed and the nucleotide sequence of three contiguous xhoi fragments, with a combined total length of approximately 3.9 kilobases, has been investigated. this region was determined to contain a total of four separate open reading frames, ea ...19901366507
molecular analysis of the hydrogenosomal ferredoxin of the anaerobic protist trichomonas vaginalis.we have determined the primary structure of the [2fe-2s]ferredoxin of the anaerobic protist trichomonas vaginalis. this protein, situated in the hydrogenosome, is composed of 93 amino acids. a comparison of t. vaginalis ferredoxin with greater than 80 other ferredoxins shows the closest similarity to [2fe-2s]putidaredoxin of the aerobic bacterium pseudomonas putida and a lesser one to mitochondrial [2fe-2s]ferredoxins of vertebrates. this similarity is reflected in the overall primary structure ...19901696716
stereochemical aspects of the oxidation of 4-ethylphenol by the bacterial enzyme 4-ethylphenol methylenehydroxylase.the o2-independent hydroxylase 4-ethylphenol methylenehydroxylase (4epmh) from pseudomonas putida jd1 catalysed the complete conversion of 4-ethylphenol into 1-(4-hydroxyphenyl)ethanol together with a small amount of 4-hydroxyacetophenone, but with no formation of the side product 4-vinylphenol reported to be formed when the similar enzyme p-cresol methylhydroxylase (pcmh) catalyses this reaction. the enantiomer of 1-(4-hydroxyphenyl)ethanol produced by 4epmh was r(+) when horse heart cytochrome ...19901697166
microbial degradation of the morphine alkaloids: identification of morphine as an intermediate in the metabolism of morphine by pseudomonas putida m10.a strain of pseudomonas putida was isolated by selective enrichment with morphine that was capable of utilising morphine as a primary source of carbon and energy for growth. experiments with whole cells showed that both morphine and codeine, but not thebaine, could be utilised. a novel nadp-dependent dehydrogenase, morphine dehydrogenase, was purified from crude cell extracts and was shown to be capable of oxidising morphine and codeine to morphinone and codeinone, respectively. this nadp-depend ...19901701625
nucleotide sequencing and characterization of pseudomonas putida catr: a positive regulator of the catbc operon is a member of the lysr family.pseudomonas putida utilizes the catbc operon for growth on benzoate as a sole carbon source. this operon is positively regulated by the catr protein, which is encoded from a gene divergently oriented from the catbc operon. the catr gene encodes a 32.2-kilodalton polypeptide that binds to the catbc promoter region in the presence or absence of the inducer cis-cis-muconate, as shown by gel retardation studies. however, the inducer is required for transcriptional activation of the catbc operon. the ...19901688844
[pseudomonas aps nutrient medium for the isolation and identification of pseudomonas aeruginosa and pseudomonas putida].pseudomonas aps selective medium has been developed on the basis of a newly detected selective antibacterial action of oxaphenamide (p-oxyphenylsalicylamide), a cholagogue. this medium permits a single-stage combined isolation and identification of p. aeruginosa after 16-24 hrs incubation of inoculated material at 42 degrees c. if the material is incubated at 35-37 degrees c, isolation of p. putida and p. aeruginosa is possible, that are differentiated by a nitroreductase microtest within 3 hrs. ...19901705609
microbial metabolism of quinoline and related compounds. v. degradation of 1h-4-oxoquinoline by pseudomonas putida 33/1.a bacterial strain was isolated with the ability to use 1h-4-oxoquinoline as the sole source of carbon, nitrogen and energy. on the basis of its physiological properties, this isolate was classified as pseudomonas putida. 1h-3-hydroxy-4-oxoquinoline, n-formylanthranilic acid, anthranilic acid and catechol were identified as intermediates in the degradation pathway. the latter was further degraded by ortho-cleavage. the enzymatic conversion of 1h-4-oxoquinoline into 1h-3-hydroxy-4-oxoquinoline re ...19901963786
mechanism of action of urocanase. specific 13c-labelling of the prosthetic nad+ and revision of the structure of its adduct with imidazolylpropionate.1. [4-13c]nicotinate was synthesised and used to support the growth of a nicotinate auxotrophic mutant of pseudomonas putida. 13c-nmr spectroscopy of the isolated urocanase confirmed the efficient incorporation of 13c into c4 of the nicotinamide ring of the tightly bound nad+ cofactor. 2. beta-[( 2'-13c]imidazol-4-yl)propionate was synthesised according to known procedures and used for inhibition of the 13c-labelled urocanase. an increase in the absorbance at 330 nm indicated adduct formation be ...19901976515
the structure of iron superoxide dismutase from pseudomonas ovalis complexed with the inhibitor azide.the 2.9 a resolution structure of iron superoxide dismutase (fesod) (ec 1.15.1.1) from pseudomonas ovalis complexed with the inhibitor azide was solved. comparison of this structure with free enzyme shows that the inhibitor is bound at the open coordination position of the iron, with a bond length of 2.0 a. the metal moves by 0.4 a into the trigonal plane to produce an orthogonal geometry at the iron. binding of the inhibitor also causes a movement of the axial ligand (histidine 26) away from th ...19902075185
degradation of the metal-cyano complex tetracyanonickelate(ii) by cyanide-utilizing bacterial isolates.ten bacterial isolates capable of growth on tetracyanonickelate(ii) [k2[ni(cn)4]] (tcn) as the sole nitrogen source were isolated from soil, freshwater, and sewage sludge enrichments. seven of the 10 were identified as pseudomonads, while the remaining 3 were classified as klebsiella species. a detailed investigation of one isolate, pseudomonas putida bcn3, revealed a rapid growth rate on tcn (generation time, 2 h), with substrate removal and growth occurring in parallel. in addition to tcn, all ...19902082819
physical map of the aromatic amine and m-toluate catabolic plasmid ptdn1 in pseudomonas putida: location of a unique meta-cleavage pathway.a restriction endonuclease map was derived for the aromatic amine and m-toluate catabolic plasmid ptdn1 present in pseudomonas putida ucc22, a derivative of p. putida mt-2. the plasmid is 79 +/- 1 kbp in size and can be divided into a restriction-site-deficient region of 51 +/- 1 kbp and a restriction-site-profuse region of 28 kbp which begins and ends with directly repeating sequences of at least 2 kbp in length. a mutant plasmid isolated after growth of the host on benzoate had lost the restri ...19902168927
loss of tdn catabolic genes by deletion from and curing of plasmid ptdn1 in pseudomonas putida: rate and mode of loss are substrate and ph dependent.the ability to degrade aromatic amines and m-toluate (tdn+ phenotype), encoded by plasmid ptdn1, was lost from pseudomonas putida hosts after subculture in benzoate, succinate, acetate and glucose minimal medium, the fastest rate of loss occurring where benzoate was the substrate. tdn- cells had either lost the entire ptdn1 plasmid or suffered a recombinational deletion of a specific 26 kbp region. proportional increase of tdn- cells resulted from their growth-rate advantage, and additionally, w ...19902168928
phylogenetic comparisons of the branched-chain alpha-ketoacid dehydrogenase complex.1. antibodies against the e1b and e2b components of bovine branched-chain alpha-ketoacid (bcka) dehydrogenase (bckad) complex completely inhibited bcka oxidation in mammalian and avian mitochondria. bcka oxidation by salmonid mitochondria was less affected and the enzyme from pseudomonas putida was unaffected. 2. in rodents, anti-e1b e2b igg inhibited oxidation of all three bcka in a similar dose-dependent manner: oxidation of alpha-ketobutyrate and alpha-keto-y-methiolbutyrate was also partiall ...19902085956
nucleotide sequence of the gyrb gene of pseudomonas putida. 19902170947
structure of the dnaa region of micrococcus luteus: conservation and variations among eubacteria.a phylogenetic tree constructed by 5s rrna analysis is composed of three major branches in eubacteria: high g + c gram+, low g + c gram+ and gram- [hori and osawa, mol. biol. evol. 4 (1987) 445-472]. we have shown that the characteristic dnaa region is common among escherichia coli (gram-), pseudomonas putida (gram-), and bacillus subtilis (low g + c gram+). we have now determined the structure of the dnaa region of micrococcus luteus, as a representative of the last branch, high g + c gram+. th ...19902172090
chlorinated biphenyl mineralization by individual populations and consortia of freshwater bacteria.comparative studies were performed to investigate the contribution of microbial consortia, individual microbial populations, and specific plasmids to chlorinated biphenyl biodegradation among microbial communities from a polychlorinated biphenyl-contaminated freshwater environment. a bacterial consortium, designated lps10, was shown to mineralize 4-chlorobiphenyl (4cb) and dehalogenate 4,4'-dichlorobiphenyl. the lps10 consortium involved three isolates: pseudomonas testosteroni (lps10a), which m ...19902117875
cloning and expression of rat histidase. homology to two bacterial histidases and four phenylalanine ammonia-lyases.histidase (histidine ammonia-lyase, ec 4.3.1.3) catalyzes the deamination of histidine to urocanic acid. apart from phenylalanine ammonia-lyase, which is not expressed in animals, histidase is the only enzyme known to have a dehydroalanine residue in its active site. the amino site precursor and the mechanism of formation of dehydroalanine are not known. as an initial step to determining the precursor of dehydroalanine in histidase, we have isolated a functional cdna clone for histidase from a r ...19902120224
identification and characterization of genes for a second anthranilate synthase in pseudomonas aeruginosa: interchangeability of the two anthranilate synthases and evolutionary implications.two anthranilate synthase gene pairs have been identified in pseudomonas aeruginosa. they were cloned, sequenced, inactivated in vitro by insertion of an antibiotic resistance gene, and returned to p. aeruginosa, replacing the wild-type gene. one anthranilate synthase enzyme participates in tryptophan synthesis; its genes are designated trpe and trpg. the other anthranilate synthase enzyme, encoded by phna and phnb, participates in the synthesis of pyocyanin, the characteristic phenazine pigment ...19902153661
microbial metabolism of quinoline and related compounds. vii. quinoline oxidoreductase from pseudomonas putida: a molybdenum-containing enzyme.the quinoline oxidoreductase from pseudomonas putida was purified 50-fold to homogeneity with 21% recovery, using ammonium sulfate precipitation, hydrophobic interaction-, anion exchange-, and gel chromatography. the mr of the native enzyme was calculated to be 300,000 by gel filtration. sds-polyacrylamide gel electrophoresis of the enzyme revealed three protein bands corresponding to mr 85,000, 30,000 and 20,000. the enzyme contained 8 atoms of iron, 8 atoms of acid-labile sulfide, 2 molecules ...19902090161
pseudomonas chromosomal replication origins: a bacterial class distinct from escherichia coli-type origins.the bacterial origins of dna replication have been isolated from pseudomonas aeruginosa and pseudomonas putida. these origins comprise a second class of bacterial origins distinct from enteric-type origins: both origins function in both pseudomonas species, and neither functions in escherichia coli; enteric origins do not function in either pseudomonad. both cloned sequences hybridize to chromosomal fragments that show properties expected of replication origins. these origin plasmids are highly ...19902106132
design of an enzymatic hybrid system: a useful strategy for the biosynthesis of benzylpenicillin in vitro.a hybrid (prokaryotic-eukaryotic) enzyme system leading to the production of benzylpenicillin has been developed. in vitro synthesis of penicillin g was achieved by incubating 6-aminopenicillanic acid, coa, phenylacetic acid, homogeneously pure phenylacetyl-coa ligase (pa-coa ligase) from pseudomonas putida and acyl-coa:6-apa acyltransferase (at) from penicillium chrysogenum. benzylpenicillin was also obtained when at was coupled with pa-coa ligase and isopenicillin n-synthetase (ipns). this is ...19902178138
enzymatic production of l-tryptophan from dl-serine and indole by a coupled reaction of tryptophan synthase and amino acid racemase.enzymatic production of l-tryptophan from dl-serine and indole by a coupled reaction of tryptophan synthase and amino acid racemase was studied. the tryptophan synthase (ec 4.2.1.20) of escherichia coli catalyzed beta-substitution reaction of l-serine into l-tryptophan and the amino acid racemase (ec 5.1.1.10) of pseudomonas putida catalyzed the racemization of d-serine simultaneously in one reactor. under optimal conditions established for l-tryptophan production, a large-scale production of l- ...19902109982
use of a novel cassette to label phenotypically a cryptic plasmid of bacillus subtilis and map loci involved in its stable maintenance.in order to facilitate studies on the maintenance of cryptic plasmids from gram-positive bacteria we have constructed a novel cassette capg1000 (5.0 kb) which carries both a selectable marker (chloramphenicol resistance from staphylococcus aureus plasmid pc194) and a screenable marker (the xyle gene from the tol plasmid of pseudomonas putida expressed from a cloned promoter of bacillus phage spo2) and which is flanked by terminators to prevent transcription from the cassette activating or inhibi ...19902116498
purification, characterization, and structure of pseudobactin 589 a, a siderophore from a plant growth promoting pseudomonas.under conditions of low-iron stress the plant growth promoting bacterium pseudomonas putida 589 (dsm 50202) produced a yellow-green fluorescent iron-binding peptide siderophore, which was designated pseudobactin 589 a and had an affinity constant toward fe3+ of 10(25) at ph 7. protonated pseudobactin 589 a had the molecular formula c54h78o26n15 and a nominal mass spectral molecular mass of 1353 g/mol. its structure was determined by a combination of nuclear magnetic resonance, fast atom bombardm ...19902145034
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