nucleotide sequencing and transcriptional mapping of the genes encoding biphenyl dioxygenase, a multicomponent polychlorinated-biphenyl-degrading enzyme in pseudomonas strain lb400.the dna region encoding biphenyl dioxygenase, the first enzyme in the biphenyl-polychlorinated biphenyl degradation pathway of pseudomonas species strain lb400, was sequenced. six open reading frames were identified, four of which are homologous to the components of toluene dioxygenase from pseudomonas putida f1 and have been named bpha, bphe, bphf, and bphg. from this comparison, biphenyl dioxygenase was found to be a multicomponent enzyme containing a two-subunit iron-sulfur protein, a ferredo ...19921569021
resonance raman investigations of escherichia coli-expressed pseudomonas putida cytochrome p450 and p420.high-resolution resonance raman spectra of the ferric, ferrous, and carbonmonoxy (co)-bound forms of wild-type escherichia coli-expressed pseudomonas putida cytochrome p450cam and its p420 form are reported. the ferric and ferrous species of p450 and p420 have been studied in both the presence and absence of excess camphor substrate. in ferric, camphor-bound, p450 (mos), the e. coli-expressed p450 is found to be spectroscopically indistinguishable from the native material. although substrate bin ...19921581294
evaluation of the 4-hour rapid nf plus method for identification of 345 gram-negative nonfermentative rods.the ability of the rapid nf plus system (innovative diagnostic systems, inc., atlanta, ga.) to identify 345 nonfermentative gram-negative rods was evaluated. kits were inoculated with no. 1 mcfarland suspensions, and reactions were interpreted after a 4-h incubation at 35 degrees c. overall, the method correctly identified 311 strains (90.1%) without additional tests and 21 strains (6.1%) with additional tests, and 13 strains (3.8%) were misidentified. five of 13 misidentified strains were alcal ...19921583129
[the destruction of diethylene glycol by a pseudomonas putida bs-2 culture]. 19921584086
nucleotide sequence of the structural gene encoding a 2-haloalkanoic acid dehalogenase of pseudomonas putida strain aj1 and purification of the encoded protein.the nucleotide sequence of a gene encoding an l-2-haloalkanoic acid halidohydrolase from pseudomonas putida strain aj1 was determined. the orf (hadl) codes for a polypeptide of 227 amino acids (mr 25,687) which has significant homology to two other l-2-haloalkanoic acid halidohydrolases of pseudomonas sp., dehci and dehcii; these show 38% and 51% amino acid identity respectively to hadl. all three enzymes produce products of an opposite optical configuration to that of the substrates. comparison ...19921588303
expression and regulation of a dnaa homologue isolated from pseudomonas putida.a gene homologous to the escherichia coli dnaa gene was isolated from pseudomonas putida and its transcription was investigated in e. coli as well as in p. putida. in both species the p. putida dnaa gene is transcribed from two promoters, one of which shows strong homology to promoters recognized by the sigma 54 factor found in both bacteria. in e. coli transcription of the p. putida dnaa gene can be repressed by overproduction of e. coli dnaa protein, presumably due to the presence of several d ...19921588913
substrate-specificity of benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase encoded by tol plasmid pww0. metabolic and mechanistic implications.the substrate-specificities of benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase, encoded by tol plasmid pww0 of pseudomonas putida mt-2, were determined. the rates of benzyl alcohol dehydrogenase-catalysed oxidation of substituted benzyl alcohols and reduction of substituted benzaldehydes were independent of the electronic nature of the substituents at positions 3 and 4. substitutions at position 2 of benzyl alcohol affected the reactivity of benzyl alcohol dehydrogenase: the velocity ...19921590768
analysis of mutations in trfa, the replication initiation gene of the broad-host-range plasmid rk2.plasmids with mutations in trfa, the gene encoding the replication initiation protein of the broad-host-range plasmid rk2, were isolated and characterized. mutants identified from a nitrosoguanidine bank were defective in supporting the replication of a wild-type rk2 origin in escherichia coli. most of the mutations were clustered in a region of trfa corresponding to the carboxy-terminal quarter of the trfa protein. 5' and 3' deletion mutants of trfa were also constructed. a c-terminal deletion ...19921597426
differential bioavailability of soil-sorbed naphthalene to two bacterial species.prediction of the fate of hydrophobic organic contaminants in soils is complicated by the competing processes of sorption and biodegradation. to test the hypothesis that sorbed naphthalene is unavailable to degradative microorganisms, we developed a simple kinetic method to examine the rates and extents of naphthalene degradation in soil-free and soil-containing systems in a comparison of two bacterial species. the method is predicated on the first-order dependence of the initial mineralization ...19921599237
isolation and screening of plasmids from the epilithon which mobilize recombinant plasmid pd10.this study examined the potential of bacteria from river epilithon to mobilize a recombinant catabolic plasmid, pd10, encoding 3-chlorobenzoate degradation and kanamycin resistance. fifty-four mobilizing plasmids were exogenously isolated by triparental matings between strains of pseudomonas putida and epilithic bacteria from the river taff (south wales, united kingdom). frequencies for mobilization ranged from 1.7 x 10(-8) to 4.5 x 10(-3) per recipient at 20 degrees c. the sizes of the mobilizi ...19921599248
crystallization and preliminary x-ray diffraction analysis of p450terp and the hemoprotein domain of p450bm-3, enzymes belonging to two distinct classes of the cytochrome p450 superfamily.cytochromes p450 are members of a superfamily of hemoproteins that are involved in the metabolism of various physiologic and xenobiotic organic compounds. this superfamily of proteins can be divided into two classes based on the electron donor proximal to the p450: an iron-sulfur protein for class i p450s or a flavoprotein for class ii. the only known tertiary structure of any of the cytochromes p450 is that of p450cam, a class i soluble enzyme isolated from pseudomonas putida (product of the cy ...19921608967
pseudomonas putida kt2442 cultivated on glucose accumulates poly(3-hydroxyalkanoates) consisting of saturated and unsaturated monomers.the biosynthesis of poly(3-hydroxyalkanoates) (phas) by pseudomonas putida kt2442 during growth on carbohydrates was studied. phas isolated from p. putida cultivated on glucose, fructose, and glycerol were found to have a very similar monomer composition. in addition to the major constituent 3-hydroxydecanoate, six other monomers were found to be present: 3-hydroxyhexanoate, 3-hydroxyoctanoate, 3-hydroxydodecanoate, 3-hydroxydodecenoate, 3-hydroxytetradecanoate, and 3-hydroxytetradecenoate. the ...19921610179
histidine ammonia-lyase from streptomyces griseus.histidine ammonia-lyase (histidase; huth) has been purified to homogeneity from streptomyces griseus and the n-terminal amino acid (aa) sequence used to clone the histidase-encoding structural gene, huth. the purified enzyme shows typical saturation kinetics and is inhibited competitively by d-histidine and histidinol phosphate. high concentrations of k.cyanide inactivate huth unless the enzyme is protected by the substrate or histidinol phosphate. on the basis of the nucleotide sequence, the hu ...19921612436
genetic analysis of the agga locus involved in agglutination and adherence of pseudomonas putida, a beneficial fluorescent isolate of pseudomonas putida, which rapidly adheres to plant roots, is agglutinated by a glycoprotein from root surfaces. agglutination is prevented and adherence to the root surface is diminished by tn5 insertion in mutant 5123. two cosmid clones from wild type p. putida and a 2.7-kbp ecori-hindiii subclone present in both cosmid clones restored agglutinable to wild type levels in transconjugants of the nonagglutinable (agg-) tn5 mutant 5123. these three clones increased agglutinability in ...19921617198
analysis of an upstream regulatory sequence required for activation of the regulatory gene xyls in xylene metabolism directed by the tol plasmid of pseudomonas putida.transcription from the promoter of a positive regulatory gene, xyls, on the tol plasmid of pseudomonas putida is activated by another positive regulator, xylr, in the presence of m-xylene and is dependent on rna polymerase containing the ntra protein (sigma 54). deletion analysis of the upstream region of the xyls gene revealed an upstream regulatory sequence (urs), located between 145 and 188 bp upstream from the transcription start site. the urs is active in either orientation and can be place ...19921620097
acyloin formation by benzoylformate decarboxylase from pseudomonas putida.whole cells and cell extracts of pseudomonas putida grown in a medium containing ammonium mandelate have the capacity to produce the acyloin compound 2-hydroxypropiophenone when incubated with benzoylformate and acetaldehyde. benzaldehyde and benzyl alcohol were formed as reaction by-products. the enantiomeric excess of the 2-hydroxypropiophenone product was found to be 91 to 92%. the absolute configuration of the enzymatically prepared product at the carbinol carbon was found to be s. the thiam ...19921622241
conversion of cis unsaturated fatty acids to trans, a possible mechanism for the protection of phenol-degrading pseudomonas putida p8 from substrate toxicity.a trans unsaturated fatty acid was found as a major constituent in the lipids of pseudomonas putida p8. the fatty acid was identified as 9-trans-hexadecenoic acid by gas chromatography, argentation thin-layer chromatography, and infrared absorption spectrometry. growing cells of p. putida p8 reacted to the presence of sublethal concentrations of phenol in the medium with changes in the fatty acid composition of the lipids, thereby increasing the degree of saturation. at phenol concentrations whi ...19921622260
naphthalene degradation via salicylate and gentisate by rhodococcus sp. strain b4.rhodococcus sp. strain b4, isolated from a soil sample contaminated with polycyclic aromatic hydrocarbons, grows with naphthalene as the sole source of carbon and energy. salicylate and gentisate were identified as intermediates in the catabolism of naphthalene. in contrast to the well-studied catabolic pathway encoded by the nah7 plasmid of pseudomonas putida, salicylate does not induce the genes of the naphthalene-degradative pathway in rhodococcus sp. strain b4. the key enzymes of naphthalene ...19921622263
molecular characterization of the entner-doudoroff pathway in escherichia coli: sequence analysis and localization of promoters for the edd-eda operon.the nucleotide sequence of the entire escherichia coli edd-eda region that encodes the enzymes of the entner-doudoroff pathway was determined. the edd structural gene begins 236 bases downstream of zwf. the eda structural gene begins 34 bases downstream of edd. the edd reading frame is 1,809 bases long and encodes the 602-amino-acid, 64,446-da protein 6-phosphogluconate dehydratase. the deduced primary amino acid sequences of the e. coli and zymomonas mobilis dehydratase enzymes are highly conse ...19921624451
characterization of the genes encoding beta-ketoadipate: succinyl-coenzyme a transferase in pseudomonas putida.beta-ketoadipate:succinyl-coenzyme a transferase (beta-ketoadipate:succinyl-coa transferase) (ec carries out the penultimate step in the conversion of benzoate and 4-hydroxybenzoate to tricarboxylic acid cycle intermediates in bacteria utilizing the beta-ketoadipate pathway. this report describes the characterization of a dna fragment from pseudomonas putida that encodes this enzyme. the fragment complemented mutants defective in the synthesis of the coa transferase, and two proteins of ...19921624453
microbial metabolism of quinoline and related compounds. xiii. purification and properties of 1h-4-oxoquinoline monooxygenase from pseudomonas putida strain 33/1.1h-4-oxoquinoline monooxygenase was purified to homogeneity from pseudomonas putida strain 33/1 which can use 1h-4-oxoquinoline as sole source of carbon and energy. the apparent m(r) of the native enzyme was determined to be 126,000 by gel chromatography. sds polyacrylamide gel electrophoresis of the enzyme revealed one protein band corresponding to m(r) 42,000. the enzyme consists of three probably identical subunits with a relative molecular mass of about 42,000. the enzyme requires oxygen and ...19921627263
characterization of pseudomonas putida mutants unable to catabolize benzoate: cloning and characterization of pseudomonas genes involved in benzoate catabolism and isolation of a chromosomal dna fragment able to substitute for xyls in activation of the tol lower-pathway promoter.mutants of pseudomonas putida mt-2 that are unable to convert benzoate to catechol were isolated and grouped into two classes: those that did not initiate attack on benzoate and those that accumulated 3,5-cyclohexadiene-1,2-diol-1-carboxylic acid (benzoate diol). the latter mutants, represents by strain pp0201, were shown to lack benzoate diol dehydrogenase (bend) activity. mutants from the former class were presumed either to carry lesions in one or more subunit structural genes of benzoate dio ...19921629155
lipoamide dehydrogenase from azotobacter vinelandii. the role of the c-terminus in catalysis and dimer stabilization.the 10 c-terminal residues are not visible in the crystal structure of lipoamide dehydrogenase from azotobacter vinelandii, but can be observed in the crystal structures of the lipoamide dehydrogenases from pseudomonas putida and pseudomonas fluorescens. in these structures, the c-terminus folds back towards the active site and is involved in interactions with the other subunit. the function of the c-terminus of lipoamide dehydrogenase from a. vinelandii was studied by deletion of 5, 9 and 14 re ...19921633805
biodegradation of mixtures of substituted benzenes by pseudomonas sp. strain js150.pseudomonas sp. strain js150 was isolated as a nonencapsulated variant of pseudomonas sp. strain js1 that contains the genes for the degradative pathways of a wide range of substituted aromatic compounds. pseudomonas sp. strain js150 grew on phenol, ethylbenzene, toluene, benzene, naphthalene, benzoate, p-hydroxybenzoate, salicylate, chlorobenzene, and several 1,4-dihalogenated benzenes. we designed experiments to determine the conditions required for induction of the individual pathways and to ...19921637161
expression of pseudomonas aeruginosa nitrite reductase in pseudomonas putida and characterization of the recombinant protein.nitrite reductase from pseudomonas aeruginosa has been successfully expressed in pseudomonas putida. the purified recombinant enzyme contains haem c but no haem d1. nonetheless, like the holoenzyme from ps. aeruginosa, it is a stable dimer (molecular mass 120 kda), and electron transfer to oxidized azurin is biphasic and follows bimolecular kinetics (k1 = 1.5 x 10(5) and k2 = 2.2 x 10(4) m-1.s-1). unlike the chemically produced apoenzyme, recombinant nitrite reductase containing only haem c is w ...19921637357
plasmids with easily excisable xyle cassettes.two new vectors containing the xyle gene (encoding catechol-2,3-dioxygenase) of pseudomonas putida were constructed that serve as the source of the xyle cassette. these vectors are based on the kanamycin-resistance-encoding plasmid, pkan18. the promoter-less xyle gene is flanked by several restriction enzyme sites that allow for easy excision of this gene in the form of a cassette containing a ribosome-binding site, 7 bp upstream from the start codon. these cassettes lack any transcriptional ter ...19921644310
the ferric-pseudobactin receptor pupa of pseudomonas putida wcs358: homology to tonb-dependent escherichia coli receptors and specificity of the protein.the initial step in the uptake of iron via ferric pseudobactin by the plant-growth-promoting pseudomonas putida strain wcs358 is binding to a specific outer-membrane protein. the nucleotide sequence of the pupa structural gene, which codes for a ferric pseudobactin receptor, was determined. it contains a single open reading frame which potentially encodes a polypeptide of 819 amino acids, including a putative n-terminal signal sequence of 47 amino acids. significant homology, concentrated in fou ...19911646376
microbial metabolism of quinoline and related compounds. viii. xanthine dehydrogenase from a quinoline utilizing pseudomonas putida strain.the xanthine dehydrogenase from pseudomonas putida 86 was purified 68-fold to homogeneity with 47% recovery. sds-polyacrylamide gel electrophoresis of the enzyme revealed two protein bands corresponding to an mr of 87,000 and 52,000. the mr of the native enzyme was calculated to 550,000 by gel chromatography. the enzyme contained 4 atoms of molybdenum, 16 atoms of iron, 16 atoms of acidlabile sulphur and 4 molecules of fad. due to the composition of the cofactors the xanthine dehydrogenase belon ...19911647164
covalent structure of the diheme cytochrome subunit and amino-terminal sequence of the flavoprotein subunit of flavocytochrome c from chromatium vinosum.the complete sequence of the 21-kda cytochrome subunit of the flavocytochrome c (fc) from the purple phototrophic bacterium chromatium vinosum has been determined to be as follows: eptaemltnncagchg thgnsvgpaspsiaqmdpmvfvevmegfksgeias timgriakgystadfekmagyfkqqtyqpakqsf dtaladtgaklhdkycekchveggkpladeedy hilagqwtpylqyamsdfreerrpmekkmaskl rellkaegdagldalfafyasqq. the sequence is the first example of a diheme cytochrome in a flavocytochrome complex. although the locations of the heme binding sites an ...19911649169
expression, localization, and functional analysis of polychlorinated biphenyl degradation genes cbpabcd of pseudomonas putida.genes of pseudomonas putida strains that are capable of degrading polychlorinated biphenyls were cloned in the plasmid vector puc19. the resultant hybrid plasmid, paw6194, contained cbpabcd genes on a 9.0-kb dna fragment that was necessary for the catabolism of polychlorinated biphenyls. these genes were further subcloned on an 8.0-kb hindiii fragment of paw540. degradation of 3-chlorobiphenyl, 2,4-dichlorobiphenyl, and 2,4,5-trichlorobiphenyl into a chloro derivative of benzoic acid was found i ...19911649578
overproduction, purification, and characterization of chlorocatechol dioxygenase, a non-heme iron dioxygenase with broad substrate tolerance.we show here that purified chlorocatechol dioxygenase from pseudomonas putida is able to oxygenate a wide range of substituted catechols with turnover numbers ranging from 2 to 29 s-1. this enzyme efficiently cleaves substituted catechols bearing electron-donating or multiple electron-withdrawing groups in an intradiol manner with kcat/km values between 0.2 x 10(7) and 1.4 x 10(7) m-1 s-1. these unique catalytic properties prompted a comparison with the related but highly specific enzymes catech ...19911649626
functional analysis of the pseudomonas putida regulatory protein catr: transcriptional studies and determination of the catr dna-binding site by hydroxyl-radical footprinting.catr, a lysr family protein, positively regulates the pseudomonas putida catbc operon, which is required for growth on benzoate as a sole carbon source. transcriptional studies show that the catr and catbc promoters are divergent and overlapping by 2 bp. a beta-galactosidase promoter probe vector was constructed to analyze expression from the catr and catbc promoters under induced and uninduced conditions. as predicted, the catbc promoter is expressed only under induced conditions, while the cat ...19911649820
purification of glucose-inducible outer membrane protein oprb of pseudomonas putida and reconstitution of glucose-specific pores.a 43,000 molecular-weight, glucose-inducible, organic acid-repressible protein (oprb) was identified in the outer membrane of pseudomonas putida. oprb was surface expressed in whole cells, had a high beta-sheet content, and was heat modifiable, as demonstrated by 125i-labeling, circular dichroism spectroscopy, and mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. oprb was extracted from outer membrane preparations by using 2% lubrol px with 10 mm edta and purified by deae-se ...19911650338
sequence of the gene (phea) encoding phenol monooxygenase from pseudomonas sp. est1001: expression in escherichia coli and pseudomonas putida.the plasmid pest1412 contains the genes, phea and pheb, encoding phenol monooxygenase (pmo) and catechol 1,2-dioxygenase (c12]), respectively. thse were originally cloned from the plasmid dna of pseudomonas sp. est1001 [kivisaar et al., plasmid 24 (1990) 25-36]. although phea and pheb are cotranscribed using the promoter sequences derived from tn4652 and the level of expression of c120 activities from pest1412 was equal both in escherichia coli and in pseudomonas putida, the level of pmo activit ...19911650730
microbial metabolism of quinoline and related compounds. x. the molybdopterin cofactors of quinoline oxidoreductases from pseudomonas putida 86 and rhodococcus spec. b1 and of xanthine dehydrogenase from pseudomonas putida 86.the bis(carboxamidomethyl) derivatives of the molybdenum cofactors in three eubacterial molybdo-iron/sulphur-flavoproteins were examined. the quinoline oxidoreductases from pseudomonas putida 86 and rhodococcus spec. b1 contain molybdopterin cytosine dinucleotide. in xanthine dehydrogenase from pseudomonas putida 86, however, only molybdopterin was found. the bis(carboxamidomethyl) derivatives of all three enzymes were treated with nucleotide pyrophosphatase, but only those of the quinoline oxid ...19911657036
genetic organization and regulation of the xylose degradation genes in streptomyces rubiginosus.the xylose isomerase (xyla) and the xylulose kinase (xylb) genes from streptomyces rubiginosus were isolated, and their nucleotide sequences were determined. the xyla and xylb genes encode proteins of 388 and 481 amino acids, respectively. these two genes are transcribed divergently from within a 114-nucleotide sequence separating the coding regions. regulation of the xyl genes in s. rubiginosus was examined by fusing their promoters to the pseudomonas putida catechol dioxygenase gene and integr ...19911657868
identification of a novel composite transposable element, tn5280, carrying chlorobenzene dioxygenase genes of pseudomonas sp. strain p51.analysis of one of the regions of catabolic plasmid pp51 which encode chlorobenzene metabolism of pseudomonas sp. strain p51 revealed that the tcba and tcbb genes for chlorobenzene dioxygenase and dehydrogenase are located on a transposable element, tn5280. tn5280 showed the features of a composite bacterial transposon with iso-insertion elements (is1066 and is1067) at each end of the transposon oriented in an inverted position. when a 12-kb hindiii fragment of pp51 containing tn5280 was cloned ...19911657878
conjugational transfer of recombinant dna in cultures and in soils: host range of pseudomonas putida tol plasmids.recombinant tol plasmid pwwo-eb62 allows pseudomonas putida to grow on p-ethylbenzoate. this plasmid can be transferred to other microorganisms, and its catabolic functions for the metabolism of alkylbenzoates are expressed in a limited number of gram-negative bacteria, including members of pseudomonad rrna group i and escherichia coli. transfer of the recombinant plasmid to erwinia chrysanthemi was observed, but transconjugants failed to grow on alkylbenzoates because they lost catabolic functi ...19911660698
nucleotide sequence of xyle from the tol pdk1 plasmid and structural comparison with isofunctional catechol-2,3-dioxygenase genes from tol, pww0 and nah7.detailed restriction and nucleotide sequence analysis of the pseudomonas putida tol plasmid pdk1 xyle gene revealed significant homology with isofunctional xyle (81.5%) and nahh (78.0%) genes from the tol pww0 and nah7 plasmids. the highest degrees of nucleotide and apparent amino acid conservation (82.2 and 86.4%, respectively) among all three genes were found to exist within a region comprising 264 nucleotides encoding the c terminus. a comparison of localized regions revealed significantly gr ...19911672868
cloning and sequencing the urocanase gene (hutu) from pseudomonas putida.a clone harbouring the entire urocanase gene (hutu) was obtained from a genomic library of pseudomonas putida using oligonucleotide probes synthesised on the basis of known flanking sequences. one subunit of urocanase consists of 556 amino acids and has a molecular mass of 60,771 da.19911677899
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
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
[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
molecular cloning of a pseudomonas paucimobilis gene encoding a 17-kilodalton polypeptide that eliminates hcl molecules from gamma-hexachlorocyclohexane.pseudomonas paucimobilis ut26 is capable of growing on gamma-hexachlorocyclohexane (gamma-hch). a genomic library of p. paucimobilis ut26 was constructed in pseudomonas putida by using the broad-host-range cosmid vector pks13. after 2,300 clones were screened by gas chromatography, 3 clones showing gamma-hch degradation were detected. a 5-kb fragment from one of the cosmid clones was subcloned into puc118, and subsequent deletion and gas chromatography-mass spectrometry analyses revealed that a ...19911718942
cloning, sequence and transcriptional analysis of the structural gene for lpd-3, the third lipoamide dehydrogenase of pseudomonas putida.the third lipoamide dehydrogenase structural gene of pseudomonas putida, lpd3, was isolated from a library of p. putida ppg2 dna cloned in escherichia coli tb1. the nucleotide sequence of lpd3 and its flanking regions indicate that lpd3 is not part of an operon, which is unique for a prokaryotic lipoamide dehydrogenase. an open reading frame was found 207 bases upstream from the start of transcription, but is encoded on the strand opposite lpd3. there is no evidence of an open reading frame imme ...19911722146
substrate recognition sites in cytochrome p450 family 2 (cyp2) proteins inferred from comparative analyses of amino acid and coding nucleotide sequences.the substrate recognition regions in cytochrome p450 family 2 (cyp2) proteins were inferred by group-to-group alignment of cyp2 sequences and those of bacterial p450s, including pseudomonas putida p450 101a (p450cam), whose substrate-binding residues have been definitely identified by x-ray crystallography of a substrate-bound form (poulos t. l., finzel, b. c., and howard, a. j. (1987) j. mol. biol. 195, 687-700). the six putative substrate recognition sites, srss, thus identified are dispersive ...19921730627
cis-diol dehydrogenases encoded by the tol pww0 plasmid xyll gene and the acinetobacter calcoaceticus chromosomal bend gene are members of the short-chain alcohol dehydrogenase the aerobic degradation of benzoate by bacteria, benzoate is first dihydroxylated by a ring-hydroxylating dioxygenase to form a cis-diol (1,2-dihydroxycyclohexa-3,4-diene carboxylate) which is subsequently transformed to a catechol by an nad(+)-dependent cis-diol dehydrogenase. the structural gene for this dehydrogenase, encoded on tol plasmid pww0 of pseudomonas putida (xyll) and that encoded on the chromosome of acinetobacter calcoaceticus (bend), were sequenced. they encode polypeptides of ...19921740120
[characteristics of the r-plasmid pm3 (incp-9) of a broad circle of hosts].a new broad host range plasmid pm3 (incp-9) was found in a facultative methylotrophic bacteria pseudomonas putida and described. the pm3 plasmid is characterized by thermo-instability in enterobacteriaceae family of bacteria at 36 degrees c or higher temperatures. it is also unable to be inherited as an autonomous element in the obligate methylotrophic bacteria methylobacillus m75. the peculiarities of plasmid inheritance make possible to use it as a tool for genetic research, for instance, to c ...19911745275
subcloning of bph genes from pseudomonas testosteroni b-356 in pseudomonas putida and escherichia coli: evidence for dehalogenation during initial attack on chlorobiphenyls.the bpha, -b, -c, and -d genes from pseudomonas testosteroni b-356 were mapped to a 5.5-kb dna fragment of cloned plasmids pda1 and pda2 by use of deletion and insertion mutants of these plasmids. the expression of each of these genes was evaluated in escherichia coli and in pseudomonas putida, and it was found that the bphc and bphd genes are well expressed in both e. coli and p. putida cells while the bpha and bphb genes are very poorly expressed in e. coli, even when placed downstream of a ta ...19911746948
septicaemia and septic arthritis due to pseudomonas putida in a neutropenic patient. 19911753153
"in vitro" synthesis of different naturally-occurring, semisynthetic and synthetic penicillins using a new and effective enzymatic coupled system.forty-seven different penicillins, including some of great clinical importance, have been synthesized "in vitro" by coupling the newly described enzyme phenylacetyl-coa ligase (pcl) from pseudomonas putida and acyl-coa: 6-aminopenicillanic acid (6-apa) acyltransferase (at) from penicillium chrysogenum. incubations were carried out at 30 degrees c in 50 mm hcl-tris buffer ph 8.0. the reaction mixtures contained 6-apa, coa, atp, dithiothreitol, mg2+ and the corresponding penicillin side-chain prec ...19911761422
synthesis of 3-furylmethylpenicillin using an enzymatic procedure.3-furylmethylpenicillin was synthesized in vitro from 3-furylacetic acid, 6-aminopenicillanic acid (6-apa), coa, atp and mg2+. the reaction was catalyzed in two steps by the enzymes phenyl-acetyl-coa ligase (pcl) from pseudomonas putida and acyl-coa: 6-apa acyltransferase (at) from penicillium chrysogenum. pcl catalyzes the activation of 3-furylacetic acid to 3-furylacetyl-coa (3-f-coa) and at acylates the amino group of 6-apa with the 3-furylacetyl moiety of 3-f-coa, releasing coa and 3-furylme ...19911778415
[cloning of genes degrading 3-chlorobenzoate from pseudomonas putida strain 87].the ability of pseudomonas putida strain 87 to catabolize 3-chlorobenzoate was shown to be mediated by genes of pbs109 plasmid. the plasmid may be transferred by conjugation into p. aeruginosa pao2175. it seems possible that the pbs109 plasmid codes for pyrocatechase ii specific for halogenated catechol, but not catechol. the genes specifying utilization of 3-chlorobenzoate from pbs109 plasmid were cloned in the 5.5 kb bgiii fragment by using broad-host cloning system. the resulting pbs110 plasm ...19911778448
biotransformation of nitrobenzene by bacteria containing toluene degradative pathways.nonpolar nitroaromatic compounds have been considered resistant to attack by oxygenases because of the electron withdrawing properties of the nitro group. we have investigated the ability of seven bacterial strains containing toluene degradative pathways to oxidize nitrobenzene. cultures were induced with toluene vapor prior to incubation with nitrobenzene, and products were identified by high-performance liquid chromatography and gas chromatography-mass spectrometry. pseudomonas cepacia g4 and ...19911781679
metabolism of and inhibition by chlorobenzoates in pseudomonas putida p111.pseudomonas putida p111 was isolated by enrichment culture on 2,5-dichlorobenzoate and was also able to grow on 2-chloro-, 3-chloro-, 4-chloro-, 2,3-dichloro-, 2,4-dichloro-, and 2,3,5-trichlorobenzoates. however, 3,5-dichlorobenzoate completely inhibited growth of p111 on all ortho-substituted benzoates that were tested. when 3,5-dichlorobenzoate was added as a cosubstrate with either 3- or 4-chlorobenzoate, cell yields and chloride release were greater than those observed from growth on either ...19911781694
anaerobic growth and cyanide synthesis of pseudomonas aeruginosa depend on anr, a regulatory gene homologous with fnr of escherichia coli.anaerobic growth of pseudomonas aeruginosa on nitrate or arginine requires the anr gene, which codes for a positive control element (anr) capable of functionally complementing an fnr mutation in escherichia coli. the anr gene was sequenced; it showed 51% identity with the fnr gene at the amino acid sequence level. four cysteine residues known to be essential in the fnr protein are conserved in anr. the anr gene product (deduced mr 27,129) was visualized by the maxicell method and migrated like a ...19911787798
microbial metabolism of quinoline and related compounds. xii. isolation and characterization of the quinoline oxidoreductase from rhodococcus spec. b1 compared with the quinoline oxidoreductase from pseudomonas putida 86.quinoline oxidoreductase from rhodococcus spec. b1 was purified 39-fold to apparent homogeneity in a 5-step procedure with a recovery of 26%. the mr of the native enzyme as determined by gel chromatography was 300,000. sds polyacrylamide gel electrophoresis of the enzyme revealed 3 protein bands corresponding to mr 82,000, 32,000, and 18,000. the enzyme contains 1.3 atoms of molybdenum, 8 atoms of iron, 8 atoms of acid-labile sulphur, 2 molecules of fad and 2 molecules of molybdopterin cytosine ...19911789933
phenotypic variation of pseudomonas putida and p. tolaasii affects the chemotactic response to agaricus bisporus mycelial exudate.the chemotactic response of wild-type pseudomonas putida and p. tolaasii, and a phenotypic variant of each species, to agaricus bisporus mycelial exudate was examined. both p. putida, the bacterium responsible for initiating basidiome development of a. bisporus, and p. tolaasii, the causal organism of bacterial blotch disease of the mushroom, displayed a positive chemotactic response to casamino acids and to a. bisporus mycelial exudate. the response was both dose- and time-dependent and marked ...19911791431
phenotypic variation of pseudomonas putida and p. tolaasii affects attachment to agaricus bisporus mycelium.the effect of phenotypic variation on attachment of pseudomonas tolaasii and p. putida to agaricus bisporus mycelium was investigated. quantitative studies demonstrated the ability of each isolate to attach rapidly and firmly to a. bisporus mycelium and significant differences in attachment of wild-type and phenotypic variant strains were observed. this was most pronounced in p. tolaasii, where the percentage attachment of the wild-type form was always greater than that of the phenotypic variant ...19911791432
isolation and characterization of pseudomonas putida mutants affected in arginine, ornithine and citrulline catabolism: function of the arginine oxidase and arginine succinyltransferase pathways.pseudomonas putida mutants impaired in the utilization of arginine are affected in either the arginine succinyltransferase pathway, the arginine oxidase route, or both. however, mutants affected in one of the pathways still grow on arginine as sole carbon source. analysis of the products excreted by both wild-type and mutant strains suggests that arginine is mainly channelled by the oxidase route. proline non-utilizing mutants are also affected in ornithine utilization, confirming the role of pr ...19911791443
dna sequence determination of the tol plasmid (pwwo) xylgfj genes of pseudomonas putida: implications for the evolution of aromatic catabolism.the meta operon of the pseudomonas putida tol plasmid (pwwo) encodes all enzymes of a meta-cleavage pathway for the metabolism of benzoic acids to krebs-cycle intermediates. we have determined and analysed the nucleic acid sequence of a 3442 bp region of the meta operon containing the xyl-gfj genes whose products are involved in the post meta-ring fission transformation of catechols. homology analysis of the xylgfj gene products revealed evidence of biochemical relatedness, suggested enzymatic m ...19911791759
[the autoselection of neustonic forms of bacteria].self-breeding of neuston forms of methylobacterium sp., pseudomonas putida bc-2, alcaligenes paradoxus bc-1, bacillus thuringiensis var. israilensis bacteria as well as of a mixed culture of methylotrophs is shown possible. in spite of ability of hydrophobicity of the cell surface the suggested method of self-breeding may be used to perfect properties of larvicidal biopreparations, and bacterial preparations which intensify self-purification of water bodies.19911791780
[quantitation of acinetobacter calcoaceticus in mixed bacterial cultures by an enzyme immunoassay].an enzyme-linked immunosorbent assay using polyclonal antibodies from rabbits has been developed for quantification of acinetobacter calcoaceticus. bacteria were added to the wells of a microtiter plate coated with anti-acinetobacter immunoglobulin. for detecting bound cells the peroxidase-labelled immunoglobulin fraction was used. over a distinct range there is a linear correlation between bound bacteria and measured absorbance allowing a quantification of bacteria in an order from 10(7) to 10( ...19911793089
highly conserved coding sequences in polychlorinated biphenyl (pcb)-degraders of pseudomonas pseudoalcaligenes kf707 and pseudomonas putida kf715.sixteen strains that utilize biphenyl or polychlorinated biphenyl (pcb) as the sole source of carbon and energy have previously been isolated. nucleotide sequence of the bphabcxd operon (11 kb) of pseudomonas pseudoalcaligenes kf707 has now been determined. when bphd region is compared with the previously reported bphd sequence of another pcb-degrader, pseudomonas putida kf715, homologies at the level of nucleotides as well as amino acids are as high as 96%. moreover, both bphabcxd operon (kf707 ...19911842046
three-dimensional structure of p-cresol methylhydroxylase (flavocytochrome c) from pseudomonas putida at 3.0-a resolution.p-cresol methylhydroxylase (pcmh) isolated from pseudomonas putida is an alpha 2 beta 2 tetramer of approximate subunit mr 49,000 and 9,000. it is a flavocytochrome c containing covalently bound fad in the larger subunit and covalently bound heme in the smaller. crystals in space group p2(1)2(1)2(1) with unit-cell parameters a = 140.3 a, b = 130.6 a, and c = 74.1 a contain one full molecule per asymmetric unit and diffract anisotropically to about 2.8-a resolution in two directions and to about ...19911846290
a series of wide-host-range low-copy-number vectors that allow direct screening for recombinants.a series of controlled expression vectors was constructed based on the wide-host-range plasmid pmmb66eh. some of these new vectors code for the alpha-peptide of beta-galactosidase and allow the direct screening of recombinant clones by inactivation of alpha-complementation. the bla gene was replaced in some plasmids by the cat gene of tn9 coding for chloramphenicol resistance, extending the use into beta-lactam-resistant strains. they all feature either the tac or taclac (tac-lac uv5 in tandem) ...19911847347
[localization of camphor degradative plasmids on the chromosome of pseudomonas putida strains paw].camphor degradative plasmids (cam, prk1) are preferentially situated on chromosomes of pseudomonas putida strains paw. after having been transferred into cam+ strains, the tol plasmid pwwo dissociates into the cryptic plasmid pwwo-8 and chromosome-borne transposon tn4651. the opposite situation, i.e. reconstruction of the tol plasmid pwwo from the cryptic plasmid pwwo-8 and chromosome-borne catabolic operons of the pwwo plasmid has been described. cam- derivatives of the cam plasmid were obtaine ...19911855659
a predicted three-dimensional structure of human cytochrome p450: implications for substrate specificity.a three-dimensional structure for human cytochrome p450ia1 was predicted based on the crystal coordinates of cytochrome p450cam from pseudomonas putida. as there was only 15% residue identity between the two enzymes, additional information was used to establish an accurate sequence alignment that is a prerequisite for model building. twelve representative eukaryotic sequences were aligned and a net prediction of secondary structure was matched against the known alpha-helices and beta-sheets of p ...19911857713
binding sites of pyridine in cytochrome p-450cam.careful titration of oxidized cytochrome p-450cam from pseudomonas putida with pyridine revealed deviations of the eadie plot from linearity in the substrate-bound as well as in the substrate-free protein. a binding model which assumes two binding sites for pyridine--the iron and the camphor binding site--is able to describe completely the nonlinear eadie plot.19911859821
hydroxylation of o-halogenophenol and o-nitrophenol by salicylate hydroxylase.salicylate hydroxylase [ec] from pseudomonas putida catalyzed the formation of catechol from substrate analogues such as o-nitro-, o-amino-, o-iodo-, o-bromo-, and o-chloro-phenol by removing the ortho-substituted groups. they are converted into nitrite, ammonia, and halide ions, respectively. kinetic parameters of these reactions were determined by spectrophotometric and polarographic methods. hydroxylation of o-nitro- or o-iodophenol proceeds with the unusual stoichiometry of 2:1:1 f ...19911864847
degradation of l-djenkolate catalyzed by s-alkylcysteine alpha,beta-lyase from pseudomonas putida.s-alkylcysteine alpha, beta-lyase [ec] of pseudomonas putida catalyzes alpha,beta-elimination of l-djenkolate [3,3'-methylenedithiobis(2-aminopropionic acid)] to produce pyruvate, ammonia, and s-(mercaptomethyl)cysteine initially. secondly, s-(mercaptomethyl)-cysteine, which was identified in the form of s-(mercaptomethyl)cysteine thiolactone and s-(2-thia-3-carboxypropyl)cysteine in the absence and presence of iodoacetic acid, respectively, is decomposed enzymatically to pyruvate, ammon ...19911869519
assay for detection and enumeration of genetically engineered microorganisms which is based on the activity of a deregulated 2,4-dichlorophenoxyacetate assay system was developed for the enumeration of genetically engineered microorganisms expressing a deregulated 2,4-dichlorophenoxyacetate (tfd) monooxygenase, which converts phenoxyacetate (paa) to phenol. in paa-amended cultures of pseudomonas aeruginosa pao1c(pro103) and pseudomonas putida ppo301(pro103), strains which express a deregulated tfd monooxygenase, phenol production was proportional to cell number. phenol was reacted, under specific conditions, with a 4-aminoantipyrine dye to f ...19911872608
cloning and characterization of an extracellular temperature-labile serine protease gene from aeromonas hydrophila.aeromonas virulence is thought to depend on multigenic functions. the gene for an extracellular protease from aeromonas hydrophila so2/2 was cloned in escherichia coli c600-1 by using pij860, bifunctional plasmid, as a vector. the gene encodes for a temperature-labile serine protease (p2) with a molecular mass of approx. 68 kda which is highly inhibited by pmsf. the gene was expressed in streptomyces lividans 1326 by transforming protoplasts with the original clone ppa2. we were also able to tra ...19911874394
activation of the pseudomonas tol plasmid upper pathway operon. identification of binding sites for the positive regulator xylr and for integration host factor protein.expression of the pseudomonas putida tol plasmid upper pathway operon requires a promoter that belongs to the -12/-24 class. stimulation of transcription from this promoter is positively controlled by the effector-activated xylr protein and requires a form of rna-polymerase holoenzyme containing the rpon-encoded sigma factor, sigma 54. xylr-dependent stimulation of transcription from the pseudomonas tol upper pathway promoter was examined using deletions, insertions, and in vivo dimethyl sulfate ...19911874736
[cloning of the arthrobacter globiformis fcba gene for dehalogenase and construction of a hybrid pathway of 4-chlorobenzoic acid degradation in pseudomonas putida].the artrobacter globiformis kzt1 fcba gene responsible for dehalogenase (4-chlorobenzoate-4-hydroxylase) activity was cloned in escherichia coli and pseudomonas putida cells. the character of the fcba gene expression was studied. notwithstanding amplification of the gene dose and control of the inducible plac promoter, the level of substrate dehalogenation by recombinant e. coli strains was lower, as compared with that in the original kzt1 strain. cloning of the fcba gene in p. putida kz6r cells ...19911879677
[genetic control of tryptophan hypersynthesis in regulatory mutants of pseudomonas putida].the 6-fluorotryptophan resistant mr1 mutant was obtained from pseudomonas putida m30 (tyr- phe-) strain. the mutant was able to excrete tryptophan (60 micrograms/ml) and has derepressed arof gene encoding 3-deoxy-d-arabinoheptulosonate-7-phosphate synthase. the mutation isolated was identified as aror with the help of cloning early arof gene of p. putida. on the next step of selection, regulatory mutant mr2 was obtained producing 240 micrograms/ml of tryptophan. the mr2 has derepressed unlinked ...19911879681
[clinical evaluation of cefpirome in children].cefpirome (cpr, hr 810) was clinically evaluated for its efficacy and safety in 11 patients with ages from 4 months to 11 years with bacterial infection. the results obtained are summarized as follows. 1. cpr was administered to 6 patients with bronchopneumonia, a patient with pneumonia, a patient with tonsillitis, 2 patients with acute pharyngitis and a patient with suppurative parotitis at daily dosage levels ranging 55.5-91.7 mg/kg, divided into 3 using intravenous bolus injection or 30 minut ...19911880923
construction of cloning cartridges for development of expression vectors in gram-negative bacteria.a cloning cartridge was constructed that can be inserted into a plasmid of choice to form an expression vector in which gene expression is inducible with an inexpensive inducer, sodium salicylate, at low concentrations. this cartridge consists of a 3.6-kb restriction fragment which contains the positive regulatory gene nahr from plasmid nah7, a promoter, pg, that nahr regulates, a multiple cloning site, a transcription terminator, and a gene conferring tetracycline resistance. within promoter pg ...19911885513
use of a xyle marker gene to monitor survival of recombinant pseudomonas putida populations in lake water by culture on nonselective media.incq marker plasmids were previously constructed to enable the analysis of the survival of populations of pseudomonas putida released into lake water (c. winstanley, j. a. w. morgan, r. w. pickup, j. g. jones, and j. r. saunders, appl. environ. microbiol. 55:771-777, 1989). we constructed equivalent incp plasmids, plv1016 and plv1017, to provide conjugative alternative systems. detection of the xyle gene carried by marker plasmids was found to be a valid indicator to use for studying the surviva ...19911892380
bacterial species dominance within a binary culture biofilm.studies with two species of bacteria, pseudomonas putida and hyphomicrobium sp. strain zv620, were carried out to evaluate the overall net rate of accumulation of biofilm, the biofilm species composition, and individual species shear-related removal rates. bacterial cells of either or both species were deposited onto glass or biofilm surfaces to initiate multispecies biofilms. subsequent biofilm development was carried out under known conditions of nutrient concentration and laminar flow. establ ...19911892387
mannopine and mannopinic acid as substrates for arthrobacter sp. strain mba209 and pseudomonas putida na513.the characteristics of mannopine and mannopinic acid utilization by agrobacterium tumefaciens b6s3, arthrobacter sp. strain mba209, and pseudomonas putida na513 were studied. strain b6s3 utilized the four mannityl opines, mannopine, mannopinic acid, agropine, and agropinic acid. it also utilized several mannityl opine analogs, which were modified in either the sugar or the amino acid moiety. it utilized mannopine more rapidly after preincubation on mannopine, mannopinic acid, or glutamine than a ...19911902209
cloning and sequence analysis of the lpd-glc structural gene of pseudomonas putida.pseudomonas putida is able to produce three lipoamide dehydrogenases: (i) lpd-glc, which is the e3 component of the pyruvate and 2-ketoglutarate dehydrogenase complexes and the l-factor for the glycine oxidation system; (ii) lpd-val, which is the specific e3 component of the branched-chain keto acid dehydrogenase complex and is induced by growth on leucine, isoleucine, or valine; and (iii) lpd-3, which was discovered in a lpdg mutant and whose role is unknown. southern hybridization with an olig ...19911902462
up-promoter mutations in the trpba operon of pseudomonas pseudomonas aeruginosa, the operon encoding tryptophan synthase (trpba) is positively regulated by the trpi protein and an intermediate in tryptophan biosynthesis, indoleglycerol phosphate (ingp). a gene fusion in which the trpba promoter directs expression of the pseudomonas putida xyle gene was constructed. by using a p. putida f1 tode mutant carrying this fusion on a plasmid, three cis-acting mutations that increased xyle expression enough to allow the tode strain to grow on toluene were i ...19911904857
cloning of pectate lyase gene pel from pseudomonas fluorescens and detection of sequences homologous to pel in pseudomonas viridiflava and pseudomonas putida.pectate lyase (pl) depolymerizes pectin and other polygalacturonates (pgas) and is thought to play a role in bacterial invasion of plants. production of pl by the soft-rotting pathogen pseudomonas fluorescens cy091 is regulated by ca2+. in the presence of ca2+, this bacterium constitutively synthesizes pl in media containing glucose, glycerol, or pga and excretes over 87% of total pl into culture fluids. in the absence of ca2+, the organism fails to use pga as a carbon source and produces very l ...19911906062
antimicrobial activity of microgard against food spoilage and pathogenic microorganisms.microgard, a commercially available fermented milk product containing antimicrobial metabolites, was a potent inhibitor for gram-negative bacteria such as pseudomonas, salmonella, and yersinia when 1% concentration was incorporated into agar media. gram-positive bacillus cereus, staphylococcus aureus, and listeria monocytogenes were insensitive to microgard. kluyveromyces marxianus, an unidentified black yeast, and penicillium expansum were partially suppressed, whereas aspergillus niger and a y ...19911906486
nmr studies on p-hydroxybenzoate hydroxylase from pseudomonas fluorescens and salicylate hydroxylase from pseudomonas putida.p-hydroxybenzoate hydroxylase from pseudomonas fluorescens and salicylate hydroxylase from pseudomonas putida have been reconstituted with 13c- and 15n-enriched fad. the protein preparations were studied by 13c-nmr, 15n-nmr and 31p-nmr techniques in the oxidized and in the two-electron-reduced states. the chemical shift values are compared with those of free flavin in water or chloroform. it is shown that the pi electron distribution in oxidized free p-hydroxybenzoate hydroxylase is comparable t ...19911915345
location and sequence of the todf gene encoding 2-hydroxy-6-oxohepta-2,4-dienoate hydrolase in pseudomonas putida f1.the gene (todf) encoding 2-hydroxy-6-oxohepta-2,4-dienoate hydrolase in pseudomonas putida f1 was shown to be located upstream of the todc1c2bade genes. the latter form part of the tod operon and encode the enzymes responsible for the initial reactions in toluene degradation. the nucleotide (nt) sequence of todf was determined and the deduced amino acid (aa) sequence revealed that the hydrolase contains 276 aa with a mr of 30,753. the deduced aa sequence was 63.5% homologous to that reported for ...19911916282
intermediate and mechanism of hydroxylation of o-iodophenol by salicylate hydroxylase.salicylate hydroxylase [ec] from pseudomonas putida catalyzes the hydroxylation of salicylate, and also o-aminophenol, o-nitrophenol, and o-halogenophenols, to catechol. the reactions with these o-substituted phenols comprise oxygenative deamination, denitration, and dehalogenation, respectively. the reaction stoichiometry, as to nadh oxidized, oxygen consumed, and catechol formed, is 2 : 1 : 1, respectively. the mechanisms for the deiodination and oxygenation of o-iodophenol were inve ...19911917904
mathematical analysis of catabolic function loss in a population of pseudomonas putida mt-2 during non-limited growth on benzoate.pseudomonas putida mt-2, harbouring the tol plasmid pww0, was grown continuously on benzoate in a phauxostat at a non-limited rate. the gradual decrease in the population carrying the complete tol plasmid was caused predominantly by a growth-rate advantage of spontaneous mutants carrying a partially deleted plasmid (tol- cells). the growth-rate difference (v) was quantified both by measuring the increase in the dilution rate (from 0.68 to 0.79 h-1; v = 0.11 h-1) and by mathematical analysis of t ...19911919510
stability of tol plasmid pww0 in pseudomonas putida mt-2 under non-selective conditions in continuous culture.pseudomonas putida mt-2, harbouring the tol plasmid pww0, was grown in chemostat culture under succinate-, sulphate-, ammonium- or phosphate-limitation at different dilution rates. the fraction of mutant cells lacking the plasmid-encoded enzymes for the degradation of toluene and xylene (tol- cells), was determined. genetic analysis revealed that all tol- cells isolated harboured partially deleted plasmids, lacking the tol catabolic genes. the growth-rate advantage of the tol- cells was quantifi ...19911919511
bioconversion of 2-hydroxy-6-oxo-6-(4'-chlorophenyl)hexa-2,4-dienoic acid, the meta-cleavage product of 4-chlorobiphenyl.bacterial conversion of 4-chlorobiphenyl (4-cb) usually proceeds through a pathway involving an initial oxidation of the unsubstituted ring in the 2,3 position followed by a 1,2 meta-cleavage. the meta-cleavage product (mcp) is converted through a single hydrolysis step into chlorobenzoic acid. however, several other acidic metabolites that were not expected as part of this pathway have already been described. in this paper, we used strains of pseudomonas putida carrying cloned genes from pseudo ...19911919512
mechanistic studies of two amino acid racemases of broad substrate specificity from pseudomonas striata and aeromonas caviae.the conversion of l-[alpha-2h]alanine in h2o and unlabeled l-alanine in 2h2o into d-alanine, under nearly irreversible conditions, with the amino acid racemase from pseudomonas striata showed significant internal transfer of the alpha-hydrogen. this result has been interpreted as being indicative of a single base mechanism for the racemization. the relative rates of deuterium incorporation into unlabeled d- and l-methionine by the two amino acid racemases of broad substrate specificity from p. s ...19911920082
[antimicrobial activity of cefpiramide to fresh clinical isolates].in the subjects of 835 strains of 37 clinically isolated microbial strains, which were separated and identified among materials collected from patients with various infections and which were sent from medical therapeutic institutions throughout japan in 1990, for the purpose of examining the antimicrobial activity of cefpiramide (cpm), its minimum inhibitory concentration (mic), together with those of other cephem antibiotics, was determined, and the following conclusions were obtained. 1. micro ...19911920808
one-step cloning system for isolation of bacterial lexa-like genes.a system to isolate lexa-like genes of bacteria directly was developed. it is based upon the fact that the presence of a lexa(def) mutation is lethal to sula+ cells of escherichia coli. this system is composed of a sula- lexa(def) hsdr- strain and a lexa-conditional killer vector (plasmid pua165) carrying the wild-type sula gene of e. coli and a polylinker in which foreign dna may be inserted. by using this method, the lexa-like genes of salmonella typhimurium, erwinia carotovora, pseudomonas ae ...19911938926
potential dna slippage structures acquired during evolutionary divergence of acinetobacter calcoaceticus chromosomal benabc and pseudomonas putida tol pww0 plasmid xylxyz, genes encoding benzoate dioxygenases.the xylxyz dna region is carried on the tol pww0 plasmid in pseudomonas putida and encodes a benzoate dioxygenase with broad substrate specificity. the dna sequence of the region is presented and compared with benabc, the chromosomal region encoding the benzoate dioxygenase of acinetobacter calcoaceticus. corresponding genes from the two biological sources share common ancestry: comparison of aligned xylx-bena, xyly-benb, and xylz-benc amino acid sequences revealed respective identities of 58.3, ...19911938949
determination of biocatalyst consumption in an aminopeptidase process using automated sample preparation and high-performance liquid chromatography.a rapid and sensitive method has been developed for the determination of the biocatalyst consumption in the chemo-enzymic production of optically pure natural and synthetic alpha-h-amino acids. it is based on automated sample preparation from an enzymic reaction mixture, reversed-phase high-performance liquid chromatographic separation, post-column reaction and fluorimetric detection. the assay procedure has been applied to the enzymic conversion of racemic norvaline amide into l-norvaline, cata ...19911939460
in vitro enzymatic synthesis of new penicillins containing keto acids as side different penicillins containing alpha-ketobutyric, beta-ketobutyric, gamma-ketovaleric, alpha-ketohexanoic, delta-ketohexanoic, epsilon-ketoheptanoic, and alpha-ketooctanoic acids as side chains have been synthesized in vitro by incubating the enzymes phenylacetyl coenzyme a (coa) ligase from pseudomonas putida and acyl-coa:6-aminopenicillanic acid acyltransferase from penicillium chrysogenum with coa, atp, mg(2+), dithiothreitol, 6-aminopenicillanic acid, and the corresponding side chain ...19911952871
ecologically significant effects of pseudomonas putida ppo301(pro103), genetically engineered to degrade 2,4-dichlorophenoxyacetate, on microbial populations and processes in soil.pseudomonas putida ppo301 (pro103), genetically engineered to degrade 2,4-dichlorophenoxyacetate, affected microbial populations and processes in a nonsterile xeric soil. in soil amended with 2,4-dichlorophenoxyacetate (500 micrograms/g soil) and inoculated with ppo301 (pro103), the rate of evolution of carbon dioxide was retarded for approximately 35 days; there was a transient increase in dehydrogenase activity; and the number of fungal propagules decreased below detection after 18 days. in un ...19911954581
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