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[evaluation of culture media used in the differentiation of pseudomonas aeruginosa, pseudomonas putida, and pseudomonas fluorescens, isolated from clinical human material]. 19744549846
pollution of modern metalworking fluids containing biocides by pathogenic bacteria in france. reexamination of chemical treatments accuracy.pollution by pathogenic bacteria was examined in 150 french metalworking fluid samples. gram-negative micro-organisms such as salmonella spp., shigella spp., and vibrio spp. as well as gram-positive cocci were never isolated. nevertheless opportunistic pathogens such as pseudomonas aeruginosa and klebsiella pneumoniae still contaminated these fluids with an isolation frequency of 17% of samples for each. these two micro-organisms failed to grow or even survive in vitro in sterile cutting fluids ...19957489767
characterization of the pilf-pild pilus-assembly locus of neisseria gonorrhoeae.expression of type iv pili by the bacterial pathogen neisseria gonorrhoeae appears to be essential for colonization of the human host. several n. gonorrhoeae gene products have been recently identified which bear homology to proteins involved in pilus assembly and protein export in other bacterial systems. we report here the isolation and characterization of transposon insertion mutants in n. gonorrhoeae whose phenotypes indicate that the n. gonorrhoeae pilf and pild gene products are required f ...19957565116
the gene encoding glyoxalase i from pseudomonas putida: cloning, overexpression, and sequence comparisons with human glyoxalase i.the gene encoding glyoxalase i (glxi) from pseudomonas putida has been cloned into the high-expression plasmid pbtaci. in the presence of iptg, jm109 cells transformed with this vector give expression levels of glxi 4000-fold higher than wild-type escherichia coli. contrary to a previous report, the nucleotide sequence of the gene encodes a 173-amino-acid polypeptide. edman analysis indicates that the predicted n-terminal methionine is lost post-translationally to yield a 19407-da protein. mass ...19947959071
[design of the human gm-csf gene using the polymerase chain reaction and its expression in pseudomonas putida cells].construction of human gm-csf gene was conducted by the pcr technique. four exons of gm-csf gene were synthesized on the basis of human blood dna using thermostable tth dna polymerase. synthetic oligonucleotides were used as primers. the oligonucleotides contained sequences complementary to the ends of exons. joining of exons was conducted by reciprocal complementation of the terminal sequences, followed by filling and amplification of the joined products. in most cases the effective synthesis of ...19947990813
isolation and sequencing of a gene coding for glyoxalase i activity from salmonella typhimurium and comparison with other glyoxalase i sequences.the glyoxalase i gene (gloa) from salmonella typhimurium has been isolated in escherichia coli on a multi-copy pbr322-derived plasmid, selecting for resistance to 3 mm methylglyoxal on luria-bertani agar. the region of the plasmid which confers the methylglyoxal resistance in e. coli was sequenced. the deduced protein sequence was compared to the known sequences of the homo sapiens and pseudomonas putida glyoxalase i (glxi) enzymes, and regions of strong homology were used to probe the national ...19979047352
multidrug efflux pumps and antimicrobial resistance in pseudomonas aeruginosa and related organisms.pseudomonas aeruginosa is an opportunistic human pathogen characterized by an innate resistance to multiple antimicrobial agents. a major contribution to this intrinsic multidrug resistance is provided by a number of broadly-specific multidrug efflux systems, including mexab-oprm and mexxy-oprm. in addition, these and two additional tripartite efflux systems, mexcd-oprj and mexef-oprn, promote acquired multidrug resistance as a result of mutational hyperexpression of the efflux genes. in additio ...200111321581
distinct classes of glyoxalase i: metal specificity of the yersinia pestis, pseudomonas aeruginosa and neisseria meningitidis enzymes.the metalloisomerase glyoxalase i (glxi) catalyses the conversion of methylglyoxal-glutathione hemithioacetal and related derivatives into the corresponding thioesters. in contrast with the previously characterized glxi enzymes of homo sapiens, pseudomonas putida and saccharomyces cerevisiae, we recently determined that escherichia coli glxi surprisingly did not display zn2+-activation, but instead exhibited maximal activity with ni2+. to investigate whether non-zn2+ activation defines a distinc ...200415270717
kinetics of 1,4-dioxane biodegradation by monooxygenase-expressing bacteria.1,4-dioxane is a probable human carcinogen, and an important emerging water contaminant. in this study, the biodegradation of dioxane by 20 bacterial isolates was evaluated, and 13 were found to be capable of transforming dioxane. dioxane served as a growth substrate for pseudonocardia dioxanivorans cb1190 and pseudonocardia benzenivorans b5, with yields of 0.09 g protein g dioxane(-1) and 0.03 g protein g dioxane(-1), respectively. cometabolic transformation of dioxane was observed for monooxyg ...200616999122
prediction of the binding site on e1 in the assembly of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus.the beta-subunit (e1beta) of the pyruvate decarboxylase (e1, alpha(2)beta(2)) component of the bacillus stearothermophilus pyruvate dehydrogenase complex was comparatively modelled based on the crystal structures of the homologous 2-oxoisovalerate decarboxylase of pseudomonas putida and homo sapiens. based on this homology modelling, alanine-scanning mutagenesis studies revealed that the negatively charged side chain of glu285 and the hydrophobic side chain of phe324 are of particular importance ...200314644451
structural basis of the dysfunctioning of human 2-oxo acid dehydrogenase complexes.2-oxo acid dehydrogenase complexes are a ubiquitous family of multienzyme systems that catalyse the oxidative decarboxylation of various 2-oxo acid substrates. they play a key role in the primary energy metabolism: in glycolysis (pyruvate dehydrogenase complex), the citric acid cycle (2-oxoglutarate dehydrogenase complex) and in amino acid catabolism (branched-chain 2-oxo acid dehydrogenase complex). malfunctioning of any of these complexes leads to a broad variety of clinical manifestations. de ...200211945122
identification of sequences encoding the detoxification metalloisomerase glyoxalase i in microbial genomes from several pathogenic organisms.the ubiquitous glyoxalase system, which is composed of two enzymes, removes cellular cytotoxic methylglyoxal (mg). in an effort to identify critical residues conserved in the evolution of the first enzyme in this system, glyoxalase i (glxi), as well as the structural implications of sequence alterations in this enzyme, a search of the national center for biotechnology information (ncbi) database of unfinished genomes was undertaken. eleven putative glxi sequences from pathogenic organisms were i ...200010824093
survival efficacy of the combination of the methioninase gene and methioninase in a lung cancer orthotopic model.we have previously demonstrated the antitumor efficacy of recombinant methioninase (rmetase) derived from pseudomonas putida. to enhance the efficacy of rmetase, we have constructed the plgfp-metsn retrovirus encoding the p. putida methioninase (met) gene fused with the green fluorescent protein (gfp) gene. plgfp-metsn or control vector plgfpsn was introduced into the human lung cancer cell line h460. the methionine level of h460-gfp-met cells was reduced to 33% of that of h460-gfp cells. rmetas ...200010770644
fulminant hepatitis complicated by small intestine infection and massive hemorrhage.a 34-year-old man diagnosed with fulminant hepatitis, caused by hepatitis b virus, and acute renal failure was referred to our hospital. after admission to the intensive care unit, the liver and renal failure were ameliorated. melena requiring transfusion occurred during the course of his illness. endoscopic examination demonstrated pseudomembranes, erosions, ulcers, and hemorrhage in the duodenum, the upper jejunum, and the terminal ileum, suggesting widespread lesions throughout the small inte ...19989658323
overexpression and large-scale production of recombinant l-methionine-alpha-deamino-gamma-mercaptomethane-lyase for novel anticancer therapy.the goal of the next generation of cancer chemotherapy is effective tumor-selectivity. a tumor-selective target with high therapeutic potential is the elevated methionine requirement of tumor cells relative to normal cells. we have termed the elevated requirement for methionine in tumors methionine dependence. to selectively target the methionine dependence of tumors for treatment on a large-scale preclinical and clinical basis, the l-methionine alpha-deamino-gamma-mercaptomethane-lyase (methion ...19979056489
a novel form of cytochrome p-450 family 4 in human polymorphonuclear leukocytes. cdna cloning and expression of leukotriene b4 omega-hydroxylase.isolation of cdna clones for human leukotriene b4 (ltb4) omega-hydroxylase clearly demonstrates that the hydroxylase is a member of the cytochrome p-450 (cyp) superfamily. cdna clones isolated from a human leukocyte cdna library with cyp4a4 cdna as a probe encode a protein of 520 amino acids with a molecular weight of 59,805. the deduced amino acid sequence contains an invariant cysteine in the conserved heme-binding domain near the c terminus, characteristic of the p-450 superfamily. the micros ...19938486631
depletion of serum methionine by methioninase in mice.methionine dependence is a tumor-specific metabolic defect found in human cancer cell lines as well as in fresh human tumor specimens. methionine dependent tumors cease growing when deprived of methionine, unlike normal cells which can substitute homocysteine for methionine for their growth requirement. we have previously purified a stable, endotoxin-free methioninase from the bacterium, pseudomonas putida. we demonstrate in this report that purified methioninase can lower the serum levels of me ...19938239522
construction and nucleotide sequence of a cdna encoding the full-length preprotein for human branched chain acyltransferase.a cdna (1.6 kilobases) for branched chain acyltransferase (e2b) isolated from a human liver library encoded only the amino-terminal half of the protein (hummel, k. b., litwer, s., bradford, a. p., aitken, a., danner, d. j., and yeaman, s. j. (1988) j. biol. chem. 263, 6165-6168). here we report the isolation of other cdnas which encode the carboxyl-terminal half of e2b and the construction of a cdna which encodes the entire pre-e2b. cdna from the original clone encoding the leader sequence, lipo ...19892708389
in vitro inhibition of human peripheral blood lymphocyte transformation by an extract of pseudomonas putida.an extract prepared from a psychrophilic strain of pseudomonas putida was found to cause a dose-dependent inhibition of [h3]tdr incorporation into human peripheral blood lymphocytes stimulated with pha, cona, pwm, or in a mixed lymphocyte reaction. the inhibition was found not to be the result of cytotoxicity, culture medium depletion of a component necessary for lymphocyte transformation, or interference with label uptake by blast lymphocytes. the extract was most effective when added prior to ...1977608686
Does compound I vary significantly between isoforms of cytochrome P450?The cytochrome P450 (CYP) enzymes are important in many areas, including pharmaceutical development. Subtle changes in the electronic structure of the active species, Compound I, have been postulated previously to account partly for the experimentally observed differences in reactivity between isoforms. Current predictive models of CYP metabolism typically assume an identical Compound I in all isoforms. Here we present a method to calculate the electronic structure and to estimate the Fe-O bond ...201121863858
efficient production of soluble recombinant single chain fv fragments by a pseudomonas putida strain kt2440 cell factory.recombinant antibody fragments have a wide range of applications in research, diagnostics and therapy. for many of these, small fragments like single chain fragment variables (scfv) function well and can be produced inexpensively in bacterial expression systems. although escherichia coli k-12 production systems are convenient, yields of different fragments, even those produced from codon-optimized expression systems, vary significantly. where yields are inadequate, alternative production systems ...201121338491
biofilms isolated from washing machines from three continents and their tolerance to a standard detergent.the goal of this comparative study was to investigate biofilm forming microorganisms living in washing machines (wms). biofilms were sampled from 11 washing machines from four countries and three continents. among the 94 isolated strains, 30% were potential human pathogens. representative strains were selected and biofilm formation was evaluated with the crystal violet (cv) assay. the majority of the wm isolates formed more biofilm than their reference strains. biofilms of p. putida wm (the larg ...201020954022
ecotoxicity and genotoxicity assessment of cytotoxic antineoplastic drugs and their metabolites.in spite of growing scientific concern about pharmaceuticals in the environment, there is still a lack of information especially with regard to their metabolites. the present study investigated ecotoxicity and genotoxicity of three widely used cytostatic agents 5-fluorouracil (5-fu), cytarabine (cyt) and gemcitabine (gemc) and their major human metabolites, i.e. alpha-fluoro-beta-alanine (fbal), uracil-1-beta-d-arabinofuranoside (arau) and 2',2'-difluorodeoxyuridine (dfdu), respectively. effects ...201020624627
broad selective efficacy of recombinant methioninase and polyethylene glycol-modified recombinant methioninase on cancer cells in vitro.elevated dependence on methionine of cancer cells is a cancer-specific metabolic defect. recombinant l-methionine alpha,gamma-lyase (rmetase), an l-methionine depleting enzyme cloned from pseudomonas putida, was shown to have efficacy on a broad series of cancer cell lines. twenty-one different human tumor cell lines (4 lung, 4 colon, 4 kidney, 4 melanoma, 3 cns, and 2 prostate) from the nci human tumor cell line screen and 7 human normal cell strains were treated with rmetase in vitro. we showe ...201020530407
bacterial ammeline metabolism via guanine deaminase.melamine toxicity in mammals has been attributed to the blockage of kidney tubules by insoluble complexes of melamine with cyanuric acid or uric acid. bacteria metabolize melamine via three consecutive deamination reactions to generate cyanuric acid. the second deamination reaction, in which ammeline is the substrate, is common to many bacteria, but the genes and enzymes responsible have not been previously identified. here, we combined bioinformatics and experimental data to identify guanine de ...201020023034
characterization of a nitroreductase with selective nitroreduction properties in the food and intestinal lactic acid bacterium lactobacillus plantarum wcfs1.nitroreductases reduce nitroaromatic compounds and other oxidants in living organisms, having interesting implications in environmental and human health. a putative nitrobenzoate reductase encoding gene (lp_0050) was recently annotated in the completed dna sequence of lactic acid bacterium lactobacillus plantarum wcfs1 strain. in this research, this l. plantarum gene was cloned and expressed, and the corresponding protein (pnba) was biochemically characterized. this l. plantarum pnba reductase i ...200919827797
heterologous production of polyketides in bacteria.polyketide natural products are among the most important microbial metabolites in human medicine and are widely used to treat both acute and degenerative diseases. the need to develop new drugs has prompted the idea of using heterologous systems for the expression of polyketide biosynthetic pathways. the basic idea behind this approach is to use heterologous bacterial systems with better growth and genetic characteristics that could support better production of a certain compound than the origin ...200919362647
studying the mechanism of rna separations using rna chromatography and its application in the analysis of ribosomal rna and rna:rna interactions.dna/rna chromatography presents a versatile platform for the analysis of nucleic acids. although the mechanism of separation of double stranded (ds) dna fragments is largely understood, the mechanism by which rna is separated appears more complicated. to further understand the separation mechanisms of rna using ion pair reverse phase liquid chromatography, we have analysed a number of dsrna and single stranded (ss) rna fragments. the high-resolution separation of dsrna was observed, in a similar ...200919155018
construction of 3d models of the cyp11b family as a tool to predict ligand binding characteristics.aldosterone is synthesised by aldosterone synthase (cyp11b2). cyp11b2 has a highly homologous isoform, steroid 11beta-hydroxylase (cyp11b1), which is responsible for the biosynthesis of aldosterone precursors and glucocorticoids. to investigate aldosterone biosynthesis and facilitate the search for selective cyp11b2 inhibitors, we constructed three-dimensional models for cyp11b1 and cyp11b2 for both human and rat. the models were constructed based on the crystal structure of pseudomonas putida c ...200717646925
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