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type v protein secretion pathway: the autotransporter story.gram-negative bacteria possess an outer membrane layer which constrains uptake and secretion of solutes and polypeptides. to overcome this barrier, bacteria have developed several systems for protein secretion. the type v secretion pathway encompasses the autotransporter proteins, the two-partner secretion system, and the recently described type vc or at-2 family of proteins. since its discovery in the late 1980s, this family of secreted proteins has expanded continuously, due largely to the adv ...200415590781
hydrolysis of nitriles using an immobilized nitrilase: applications to the synthesis of methionine hydroxy analogue derivatives.mild and selective hydrolysis of a large range of nitriles leading to carboxylic acids was achieved under neutral conditions by an immobilized and genetically modified enzyme preparation from alcaligenes faecalis atcc8750. this immobilized nitrilase has been shown to be an effective catalyst for the stereoselective hydrolysis of mandelonitrile 1a to r-(-)-mandelic acid 1c. this method is particularly useful for the production of hydroxy analogues of methionine derivatives 2c-4c that could have a ...200415612811
genetic diversity and horizontal transfer of genes involved in oxidation of reduced phosphorus compounds by alcaligenes faecalis wm2072.enrichment was performed to isolate organisms that could utilize reduced phosphorus compounds as their sole phosphorus sources. one isolate that grew well with either hypophosphite or phosphite was identified by 16s rrna gene analysis as a strain of alcaligenes faecalis. the genes required for oxidation of hypophosphite and phosphite by this organism were identified by using transposon mutagenesis and include homologs of the ptxd and htxa genes of pseudomonas stutzeri wm88, which encode an nad-d ...200515640200
pancreatic abscess secondary to alcaligenes faecalis.we report a patient with pancreatic abscesses and necrosis secondary to alcaligenes faecalis infection. he initially presented with alcohol-induced acute pancreatitis. twenty days after the initial presentation, he re-presented with increasing pain and was found to have pancreatic necrosis and abscesses. treatment was initiated with meropenem. because of persistent fevers, computed tomography-guided drainage was performed. the fluid grew a faecalis resistant to meropenem and the patient continue ...200515654182
complete conversion of nitrate into dinitrogen gas in co-cultures of denitrifying bacteria.in the past 10 years many molecular aspects of microbial nitrate reduction have been elucidated, but the ecophysiology of this process is hardly understood. in this contribution, our efforts to study the complex microbial communities and interactions involved in the reduction of nitrate to dinitrogen gas are summarized. the initial work concentrated on emission of the greenhouse gas nitrous oxide during incomplete denitrification by alcaligenes faecalis. as more research methods became available ...200515667308
genes and enzymes involved in bacterial oxidation and reduction of inorganic arsenic. 200515691908
propionate oxidation by and methanol inhibition of anaerobic ammonium-oxidizing bacteria.anaerobic ammonium oxidation (anammox) is a recently discovered microbial pathway and a cost-effective way to remove ammonium from wastewater. anammox bacteria have been described as obligate chemolithoautotrophs. however, many chemolithoautotrophs (i.e., nitrifiers) can use organic compounds as a supplementary carbon source. in this study, the effect of organic compounds on anammox bacteria was investigated. it was shown that alcohols inhibited anammox bacteria, while organic acids were convert ...200515691967
genomic analysis of anaerobic respiration in the archaeon halobacterium sp. strain nrc-1: dimethyl sulfoxide and trimethylamine n-oxide as terminal electron acceptors.we have investigated anaerobic respiration of the archaeal model organism halobacterium sp. strain nrc-1 by using phenotypic and genetic analysis, bioinformatics, and transcriptome analysis. nrc-1 was found to grow on either dimethyl sulfoxide (dmso) or trimethylamine n-oxide (tmao) as the sole terminal electron acceptor, with a doubling time of 1 day. an operon, dmsreabcd, encoding a putative regulatory protein, dmsr, a molybdopterin oxidoreductase of the dmso reductase family (dmseabc), and a ...200515716436
bacterial zoonoses and infective endocarditis, algeria.blood culture-negative endocarditis is common in algeria. we describe the etiology of infective endocarditis in this country. samples from 110 cases in 108 patients were collected in algiers. blood cultures were performed in algeria. serologic and molecular analysis of valves was performed in france. infective endocarditis was classified as definite in 77 cases and possible in 33. causative agents were detected by blood cultures in 48 cases. all 62 blood culture-negative endocarditis cases were ...200515752438
a role for fungal {beta}-glucans and their receptor dectin-1 in the induction of autoimmune arthritis in genetically susceptible mice.a combination of genetic and environmental factors can cause autoimmune disease in animals. skg mice, which are genetically prone to develop autoimmune arthritis, fail to develop the disease under a microbially clean condition, despite active thymic production of arthritogenic autoimmune t cells and their persistence in the periphery. however, in the clean environment, a single intraperitoneal injection of zymosan, a crude fungal beta-glucan, or purified beta-glucans such as curdlan and laminari ...200515781585
suppression of neutrophil accumulation in mice by cutaneous application of geranium essential oil.background: previous studies suggested that essential oils suppressed the adherence response of human neutrophils in vitro and that intraperitoneal application of geranium oil suppressed the neutrophil accumulation into peritoneal cavity in vivo. usually, essential oils are applied through skin in aromatherapy in inflammatory symptoms. the purpose of this study is to assess the effects of cutaneous application of essential oils on the accumulation of neutrophils in inflammatory sites in skin of ...200515813994
metallo-beta-lactamases: the quiet before the storm?the ascendancy of metallo-beta-lactamases within the clinical sector, while not ubiquitous, has nonetheless been dramatic; some reports indicate that nearly 30% of imipenem-resistant pseudomonas aeruginosa strains possess a metallo-beta-lactamase. acquisition of a metallo-beta-lactamase gene will invariably mediate broad-spectrum beta-lactam resistance in p. aeruginosa, but the level of in vitro resistance in acinetobacter spp. and enterobacteriaceae is less dependable. their clinical significan ...200515831827
pro-sequence and ca2+-binding: implications for folding and maturation of ntn-hydrolase penicillin amidase from e. coli.penicillin amidase (pa) is a bacterial periplasmic enzyme synthesized as a pre-pro-pa precursor. the pre-sequence mediates membrane translocation. the intramolecular pro-sequence is expressed along with the a and b chains but is rapidly removed in an autocatalytic manner. in extensive studies we show here that the pro-peptide is required for the correct folding of pa. pro-pa and pa unfold via a biphasic transition that is more pronounced in the case of pa. according to size-exclusion chromatogra ...200515843029
genetic environment and expression of the extended-spectrum beta-lactamase blaper-1 gene in gram-negative bacteria.the genetic location of the gene coding for the expanded-spectrum beta-lactamase per-1 was analyzed in a series of gram-negative isolates. it was identified as part of a composite transposon bracketed by two novel insertion elements, ispa12 and ispa13, belonging to the is4 family that possess transposases that share 63% amino acid identity and that are chromosomally located in pseudomonas aeruginosa, providencia stuartii, and acinetobacter baumannii. on the contrary, the bla(per-1) gene was iden ...200515855485
characteristics and adaptability of iron- and sulfur-oxidizing microorganisms used for the recovery of metals from minerals and their concentrates.microorganisms are used in large-scale heap or tank aeration processes for the commercial extraction of a variety of metals from their ores or concentrates. these include copper, cobalt, gold and, in the past, uranium. the metal solubilization processes are considered to be largely chemical with the microorganisms providing the chemicals and the space (exopolysaccharide layer) where the mineral dissolution reactions occur. temperatures at which these processes are carried out can vary from ambie ...200515877814
sensing nitrite through a pseudoazurin-nitrite reductase electron transfer relay.nitrite is converted to nitric oxide by haem or copper-containing enzymes in denitrifying bacteria during the process of denitrification. in designing an efficient biosensor, this enzymic turnover must be quantitatively assessed. the enzyme nitrite reductase from alcaligenes faecalis contains a redox-active blue copper centre and a nonblue enzyme-active copper centre. it can be covalently tethered to modified gold-electrode surfaces in configurations in which direct electron transfer is possible ...200515900523
shared genotypes of achromobacter xylosoxidans strains isolated from patients at a cystic fibrosis rehabilitation center.during a study examining transmission of pseudomonas aeruginosa among 76 cystic fibrosis patients in a rehabilitation center, where patients stay in close contact during prolonged periods, several clusters of patients carrying genotypically identical p. aeruginosa, as well as two clusters of 4 and 10 patients, respectively, colonized with genotypically identical achromobacter xylosoxidans strains, were discovered.200515956444
[constitutive expression and purification of alcaligenes faecalis penicillin g acylase in escherichia coli].considering alcaligenes faecalis pencillin g acylase(afpga), which possesses the attractive characteristics for beta-lactam antibiotics conversions, the gene of pga was cloned into an expressing vector pkkfpga. the recombinant plasmid contained multicopy replicon(cole 1), trc promoter, afpga gene, rrnb transcript terminator and ampicillin marker transformed escherichia coli dh5alpha. as both the recombinant plasmid and the host dh5alpha had no laclq gene, the trc promoter was always active and t ...200415974000
enzymatic transformation of bacterial polyhydroxyalkanoates into repolymerizable oligomers directed towards chemical recycling.the enzymatic transformation into an oligomer was carried out with the objective of developing the chemical recycling of bacterial polyesters. poly(r-3-hydroxyalkanoate)s (phas), such as poly[(r-3-hydroxybutyrate)-co-12%(r-3-hydroxyhexanoate)] and poly[(r-3-hydroxybutyrate)-co-12%(r-3-hydroxyvalerate)], were degraded by granulated candida antarctica lipase b immobilized on hydrophilic silica (lipase gca) in a diluted organic solvent at 70 degrees c. the degradation products were cyclic oligomers ...200515988790
alcaligenes faecalis kw-a biofilm for denitrification of nitrate-rich effluent.alcaligenes faecalis kw-a selected for possessing good denitrification efficiency was used for biofilm development. the biofilm could be developed on a glass surface within 12 hr when 5%, ix 10(8) cells/ml was used as inoculum. the microcolonies were seen in 6 hr and glycocalyx in 9 hr stage. at 24 hr the biofilm was developed fully and hence was visualised as dense mass. the biofilm protein content showed 48.5% increase in shake flask than in static condition. the exopoplymer is produced in lar ...200515991580
purification of siderophores of alcaligenes faecalis on amberlite xad.studies on siderophore production using alcaligenes faecalis bccm id 2374 revealed hydroxamate and catecholate type of siderophores at 347 microg ml-1. these fractions were purified on amberlite xad-4 column, which resulted in the separation of two bands having absorption maxima at 264 and 224 nm. the amount of pure siderophore obtained in powdered form from first and second fraction was 297 and 50 microg ml-1 respectively.200615993053
[influence of nitrogen source nh4 cl concentration on curdlan production in alcaligenes faecalis].the effect of initial ammonium chloride level on production of curdlan in alcaligenesfaecalis was investigated. it was found that ammonium chloride was the limiting substrate for cell growth during the batch fermentation process. however, the cell growth and curdlan production could not be enhanced by solely increasing the initial ammonium chloride level. the ph drop in the broth due to the consumption of ammonium chloride also effected the cell growth and curdlan production. by simultaneously i ...200516013500
ligand loop effects on the free energy change of redox and ph-dependent equilibria in cupredoxins probed on amicyanin variants.in this work, we have determined the thermodynamic parameters of the reduction of four different variants of thiobacillus versutus amicyanin by electrochemical techniques. in addition, the thermodynamic parameters were determined of the low-ph conformational change involving protonation of the c-terminal histidine ligand and the concomitant dissociation of this histidine from the cu(i) ion. in these variants, the native c-terminal loop containing the cys, his, and met copper ligands has been rep ...200516026167
tn5393d, a complex tn5393 derivative carrying the per-1 extended-spectrum beta-lactamase gene and other resistance determinants.in alcaligenes faecalis fl-424/98, a clinical isolate that produces the per-1 extended-spectrum beta-lactamase, the bla(per-1) gene was found to be carried on a 44-kb nonconjugative plasmid, named pfl424, that was transferred to escherichia coli by electroporation. investigation of the genetic context of the bla(per-1) gene in pfl424 by means of a combined cloning and pcr mapping approach revealed that the gene is associated with a transposonlike element of the tn3 family. this 14-kb element is ...200516048938
release of an enantioselective nitrilase from alcaligenes faecalis mtcc 126: a comparative study.nitrilases constitute an important class of hydrolases, however, cheap and ready availability of enzyme sources limit their practical synthetic applications. the present investigation was directed to compare the applicability of various physical cell disintegration methods namely, solid shear, liquid shear and sonication, for the release of an enantioselective nitrilase from alcaligenes faecalis mtcc 126. different parameters associated with each method were optimized in order to ensure maximal ...200516080003
calorimetric and spectroscopic investigations of the thermal denaturation of wild type nitrite reductase.nitrite reductase (nir) is a multicopper protein, with a trimeric structure containing two types of copper site: type 1 is present in each subunit whereas type 2 is localized at the subunits interface. the paper reports on the thermal behaviour of wild type nir from alcaligenes faecalis s-6. the temperature-induced changes of the copper centres are characterized by optical spectroscopy and electron paramagnetic resonance spectroscopy, and by establishing the thermal stability by differential sca ...200516085470
nitrification and denitrification in high-strength ammonium by alcaligenes faecalis.alcaligenes faecalis sp. no. 4, that has the ability of heterotrophic nitrification and aerobic denitrification in high-strength ammonium at about 1200 mg-n/l, converted about one-half of removed nh4+-n to intracellular nitrogen and nitrified only 3% of the removed nh4+. from the nitrogen balance, 40-50% of removed nh4+-n was estimated to be denitrified. production of n2 was confirmed by gc-ms and 90% of denitrified products was n2. the maximum ammonium removal rate, 29 mg-n/l h and its denitrif ...200516086258
atomic resolution structures of resting-state, substrate- and product-complexed cu-nitrite reductase provide insight into catalytic mechanism.copper-containing nitrite reductases catalyze the reduction of nitrite to nitric oxide (no), a key step in denitrification that results in the loss of terrestrial nitrogen to the atmosphere. they are found in a wide variety of denitrifying bacteria and fungi of different physiology from a range of soil and aquatic ecosystems. structural analysis of potential intermediates in the catalytic cycle is an important goal in understanding enzyme mechanism. using "crystal harvesting" and substrate-soaki ...200516093314
alcaligenes faecalis subsp. phenolicus subsp. nov. a phenol-degrading, denitrifying bacterium isolated from a graywater bioprocessor.a gram (-) coccobacillary bacterium, j(t), was isolated from a graywater bioprocessor. 16s rrna and biochemical analysis has revealed strain j(t) closely resembles alcaligenes faecalis atcc 8750t and a. faecalis subsp. parafaecalis dsm 13975t, but is a distinct, previously uncharacterized isolate. strain j(t), along with the type strain of a. faecalis and its previously described subspecies share the ability to aerobically degrade phenol. the degradation rates of phenol for strain j(t) and refer ...200516094869
biofilm reactors for industrial bioconversion processes: employing potential of enhanced reaction rates.this article describes the use of biofilm reactors for the production of various chemicals by fermentation and wastewater treatment. biofilm formation is a natural process where microbial cells attach to the support (adsorbent) or form flocs/aggregates (also called granules) without use of chemicals and form thick layers of cells known as "biofilms." as a result of biofilm formation, cell densities in the reactor increase and cell concentrations as high as 74 gl(-1) can be achieved. the reactor ...200516122390
postoperative endophthalmitis due to an unusual pathogen: alcaligenes faecalis. 200616138111
isolation and characterization of a chlorpyrifos and 3,5,6-trichloro-2-pyridinol degrading bacterium.a bacterium, isolated from contaminated soils around a chemical factory and named strain dsp3 was capable of biodegrading both chlorpyrifos and 3,5,6-trichloro-2-pyridinol. based on the results of phenotypic features, phylogenetic similarity of 16s rrna gene sequences, dna g+c content, and dna homology between strain dsp3 and reference strains, strain dsp3 was identified as alcaligenes faecalis. chlorpyrifos was utilized as the sole source of carbon and phosphorus by strain dsp3. we examined the ...200516143458
intestinal protozoa and associated bacteria in captive houbara bustards (chlamydotis undulata) in the united arab emirates.a protozoal survey was carried out in 114 captive houbara bustards (chlamydotis undulata) in the united arab emirates. trichomonas gallinarum, chilomastix gallinarum, giardia spp. and lophomonas spp. were detected from faecal samples of 49 (43%) captive houbara bustards. culture and identification studies were carried out to assess associated bacteria. the bacteria isolated from birds negative to protozoa included enterococcus spp., aerococcus spp., micrococci spp., proteus spp., enterobacter sp ...199916147554
dissimilatory iron reduction and odor indicator abatement by biofilm communities in swine manure microcosms.animal waste odors arising from products of anaerobic microbial metabolism create community relations problems for livestock producers. we investigated a novel approach to swine waste odor reduction: the addition of fecl(3), a commonly used coagulant in municipal wastewater treatment, to stimulate degradation of odorous compounds by dissimilatory iron-reducing bacteria (dirb). two hypotheses were tested: (i) fecl(3) is an effective source of redox-active ferric iron (fe(3+)) for dissimilatory re ...200516151075
new chitosan-degrading strains that produce chitosanases similar to choa of mitsuaria chitosanitabida.the betaproteobacterium mitsuaria chitosanitabida (formerly matsuebacter chitosanotabidus) 3001 produces a chitosanase (choa) that is classified in glycosyl hydrolase family 80. while many chitosanase genes have been isolated from various bacteria to date, they show limited homology to the m. chitosanitabida 3001 chitosanase gene (choa). to investigate the phylogenetic distribution of chitosanases analogous to choa in nature, we identified 67 chitosan-degrading strains by screening and investiga ...200516151097
presence of two different active nirs nitrite reductase genes in a denitrifying thauera sp. from a high-nitrate-removal-rate reactor.the nirs nitrite reductase genes were studied in two strains (strains 27 and 28) isolated from two denitrifying reactors and characterized as thauera according to their 16s rrna gene sequences. strain 28 contains a single nirs sequence, which is related to the nirs of thauera mechernichensis, and strain 27 contains two nirs sequences; one is similar to the nirs sequence from thauera mechernichensis (gene 2), but the second one (gene 8) is from a separate clade with nirs from pseudomonas stutzeri ...200516151169
antibacterial activity of essential oils from australian native plants.to date, of the australian essential oils, only tea tree (melaleuca alternifolia) and eucalyptus spp. have undergone extensive investigation. in this study a range of australian essential oils, including those from anethole anisata, callistris glaucophyllia, melaleuca spp. and thyptomine calycina, were assayed for in vitro antibacterial activity. m. alternifolia was also included for comparison purposes. activity was determined using standard disc diffusion assays with each oil assayed at 100%, ...200516161028
characteristics of ammonium removal by heterotrophic nitrification-aerobic denitrification by alcaligenes faecalis no. 4.alcaligenes faecalis no. 4 has heterotrophic nitrification and aerobic denitrification abilities. by taking the nitrogen balance under different culture conditions, 40-50% of removed nh4+-n was denitrified and about one-half of removed nh4+-n was converted to intracellular nitrogen. the maximum ammonium removal rate of no. 4 (28.9 mg-n/l/h) and its denitrification rate at high-strength nh4+-n of about 1200 ppm in aerated batch experiments at a c/n ratio of 10 were 5-40 times higher than those of ...200516198262
use of shell-vial cell culture assay for isolation of bacteria from clinical specimens: 13 years of experience.the shell-vial culture assay is performed routinely in our laboratory. recently we revisited our experience of using the shell-vial culture assay for the isolation of microorganisms from various clinical samples. over a 13-year period, we have isolated 580 bacterial strains (5%) from 11,083 clinical samples tested. over the same period, 285 isolates of rickettsiae, bartonellae, or coxiella burnetii were cultured from a total of 7,102 samples tested. these isolates include 55 rickettsia sp. isola ...200516207953
evaluation of a rapid direct assay for identification of bacteria and the mec a and van genes from positive-testing blood cultures.we performed the first evaluation of a dna strip assay (genotype blood culture; hain lifescience, nehren, germany) for the detection of the most relevant bacterial sepsis pathogens directly from positive bactec blood culture bottles (becton dickinson, heidelberg, germany). the test comprises two panels, one for the direct species identification of important gram-positive cocci and the other for gram-negative rods. additionally, detection of the mec a and the van genes are implemented. the genoty ...200516207992
extended-spectrum beta-lactamases: a clinical update.extended-spectrum beta-lactamases (esbls) are a rapidly evolving group of beta-lactamases which share the ability to hydrolyze third-generation cephalosporins and aztreonam yet are inhibited by clavulanic acid. typically, they derive from genes for tem-1, tem-2, or shv-1 by mutations that alter the amino acid configuration around the active site of these beta-lactamases. this extends the spectrum of beta-lactam antibiotics susceptible to hydrolysis by these enzymes. an increasing number of esbls ...200516223952
influence of growth phase on bacterial cell electrokinetic characteristics examined by soft particle electrophoresis theory.the influence of incubation time on the electrokinetic properties of escherichia coli, pseudomonas putida, alcaligenes faecalis, and alcaligenes sp., was examined by electrophoretic mobility measurements and the results were discussed based on ohshima's soft particle theory. the electrophoretic mobility of e. coli plotted against incubation time revealed that the mobility gradually increased from the outset of incubation to 7 h, which corresponded to the exponential growth and early stationary p ...200316256681
analytical verification of a pcr assay for identification of bordetella avium.bordetella avium is the etiologic agent of turkey coryza or bordetellosis, a respiratory disease responsible for substantial economic losses to the turkey industry. at present, identification of this bacterium relies on isolation and biochemical testing. although a pcr for the detection of b. avium was proposed a number of years ago, lack of analytical verification precludes its use as a diagnostic tool. furthermore, a number of details pertaining to the reaction conditions used are missing or u ...200516272488
tryptophan tryptophylquinone cofactor biogenesis in the aromatic amine dehydrogenase of alcaligenes faecalis. cofactor assembly and catalytic properties of recombinant enzyme expressed in paracoccus denitrificans.the heterologous expression of tryptophan trytophylquinone (ttq)-dependent aromatic amine dehydrogenase (aadh) has been achieved in paracoccus denitrificans. the aaubeda genes and orf-2 from the aromatic amine utilization (aau) gene cluster of alcaligenes faecalis were placed under the regulatory control of the mauf promoter from p. denitrificans and introduced into p. denitrificans using a broad-host-range vector. the physical, spectroscopic and kinetic properties of the recombinant aadh were i ...200516279953
alcaligenes aquatilis sp. nov., a novel bacterium from sediments of the weser estuary, germany, and a salt marsh on shem creek in charleston harbor, usa.four nitrite-dissimilating strains, isolated from weser estuary sediments, were investigated using a polyphasic taxonomic approach. phylogenetic analysis based on 16s rrna gene sequences indicated that these strains belong to the 'betaproteobacteria' and are related to the genus alcaligenes. the highest level of sequence similarity (100 %) was found with strain m3a (=atcc 700596), a dimethyl sulfide-producing marine isolate that was included in this study. dna-dna hybridizations between the five ...200516280529
the substrate-bound type 2 copper site of nitrite reductase: the nitrogen hyperfine coupling of nitrite revealed by pulsed epr.a pulsed electron paramagnetic resonance study has been performed on the type 2 copper site of nitrite reductase (nir) from alcaligenes faecalis. the h145a mutant, in which histidine 145 is replaced by alanine, was studied by eseem and hyscore experiments at 9 ghz on frozen solutions. this mutant contains a reduced type 1 copper site which allowed a selective investigation of the type 2 site of h145a and of its nitrite-bound form h145a (no2(-)). the experiments yielded hyperfine and quadrupole p ...200516285722
l-cysteate sulpho-lyase, a widespread pyridoxal 5'-phosphate-coupled desulphonative enzyme purified from silicibacter pomeroyi dss-3(t).quantitative utilization of l-cysteate (2-amino-3-sulphopropionate) as the sole source of carbon and energy for growth of the aerobic, marine bacterium silicibacter pomeroyi dss-3(t) was observed. the sulphonate moiety was recovered in the medium largely as sulphite, and the appropriate amount of the ammonium ion was also observed. genes [suyab (3-sulpholactate sulpho-lyase)] encoding the known desulphonation reaction in cysteate degradation were absent from the genome, but a homologue of a puta ...200616302849
novel cyclic lipodepsipeptide from pseudomonas syringae pv. lachrymans strain 508 and syringopeptin antimicrobial activities.the syringopeptins are a group of antimicrobial cyclic lipodepsipeptides produced by several plant-associated pseudomonads. a novel syringopeptin, sp508, was shown to be produced as two homologs (a and b) by pseudomonas syringae pv. lachrymans strain 508 from apple and to structurally resemble syringopeptin sp22. sp508 differed from sp22 and other syringopeptins by having three instead of four alpha,beta-unsaturated amino acids and a longer beta-hydroxy acyl chain. both sp508 and sp22 displayed ...200516304170
class 1 integrons and tetracycline resistance genes in alcaligenes, arthrobacter, and pseudomonas spp. isolated from pigsties and manured soil.the presence of tetracycline resistance (tc(r)) genes and class i integrons (in-1), and their ability to cotransfer were investigated in tc(r) gram-negative (185 strains) and gram-positive (72 strains) bacteria from danish farmland and pigsties. the isolates belonged to the groups or species escherichia coli, enterobacter spp., arthrobacter spp., alcaligenes spp., pseudomonas spp., and corynebacterium glutamicum. the 257 isolates were screened for in-1. eighty-one of the gram-negative isolates w ...200516332771
tem-21 extended-spectrum beta-lactamase in a clinical isolate of alcaligenes faecalis from a nursing home. 200616344284
rapid screen for bacteria degrading water-insoluble, solid hydrocarbons on agar plates.a rapid procedure was devised for detecting on solid media bacteria able to degrade water-insoluble, solid hydrocarbons such as the polycyclic aromatic hydrocarbons phenanthrene, anthracene, and biphenyl. after alcaligenes faecalis afk2 was inoculated on a plate containing mineral salts agar, an ethereal solution of phenanthrene (about 10%, wt/vol) was sprayed on the surface of the plate, and the plate was incubated at 30 degrees c for 2 to 3 days. colonies showing degradation were surrounded wi ...198216345951
relative rates of nitric oxide and nitrous oxide production by nitrifiers, denitrifiers, and nitrate respirers.biogenic emissions of nitric and nitrous oxides have important impacts on the photochemistry and chemistry of the atmosphere. although biogenic production appears to be the overwhelming source of n(2)o, the magnitude of the biogenic emission of no is very uncertain. in soils, possible sources of no and n(2)o include nitrification by autotrophic and heterotrophic nitrifiers, denitrification by nitrifiers and denitrifiers, nitrate respiration by fermenters, and chemodenitrification. the availabili ...198616347068
microbiological degradation of malodorous substances of swine waste under aerobic conditions.phenol, p-cresol, and volatile fatty acids (vfa; acetic, propionic, isobutyric, butyric, isovaleric, and valeric acids) were used as odor indicators of swine waste. aeration of the waste allowed the indigenous microorganisms to grow and degrade these malodorous substances. the time required for degradation of these substances varied according to the waste used, and it was not necessarily related to their concentrations. using a minimal medium which contained one of the malodorous compounds as so ...198716347254
purification and characterization of d-aminoacylase from alcaligenes faecalis da1.a d-aminoacylase from alcaligenes faecalis da1 has been purified to homogeneity by a simple purification procedure with two columns, fractogel deae-650 and hw-50. the specific activity of the purified enzyme was found to be 580 u/mg of protein with n-acetyl-dl-methionine as the reaction substrate. the apparent molecular weight and isoelectric point of this enzyme were determined to be 55,000 and 5.4, respectively.199116348465
differentiation of gram-negative, nonfermentative bacteria isolated from biofilters on the basis of fatty acid composition, quinone system, and physiological reaction profiles.gram-negative, nonfermentative bacteria isolated from biofilters for off-gas treatment of animal-rendering-plant emissions were differentiated by whole-cell fatty acid analysis, quinone analysis, and numerical taxonomy based on their physiological reaction profiles. the last system consisted of 60 physiological tests and was arranged as a microtest system on microtitration plates. based on fatty acid analyses, 31 isolates were separated into six clusters and five single-member clusters. the isol ...199216348724
nitrospira-like bacteria associated with nitrite oxidation in freshwater aquaria.oxidation of nitrite to nitrate in aquaria is typically attributed to bacteria belonging to the genus nitrobacter which are members of the alpha subdivision of the class proteobacteria. in order to identify bacteria responsible for nitrite oxidation in aquaria, clone libraries of rrna genes were developed from biofilms of several freshwater aquaria. analysis of the rdna libraries, along with results from denaturing gradient gel electrophoresis (dgge) on frequently sampled biofilms, indicated the ...199816349486
purification and properties of a polyester polyurethane-degrading enzyme from comamonas acidovorans tb-35.a polyester polyurethane (pur)-degrading enzyme, pur esterase, derived from comamonas acidovorans tb-35, a bacterium that utilizes polyester pur as the sole carbon source, was purified until it showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page). this enzyme was bound to the cell surface and was extracted by addition of 0.2% n,n-bis(3-d-gluconamidopropyl)deoxycholamide (deoxy-bigchap). the results of gel filtration and sds-page showed that the pur esteras ...199816349494
ligands for the beta-glucan receptor, dectin-1, assigned using "designer" microarrays of oligosaccharide probes (neoglycolipids) generated from glucan polysaccharides.dectin-1 is a c-type lectin-like receptor on leukocytes that mediates phagocytosis and inflammatory mediator production in innate immunity to fungal pathogens. dectin-1 lacks residues involved in calcium ligation that mediates carbohydrate-binding by classical c-type lectins; nevertheless, it binds zymosan, a particulate beta-glucan-rich extract of saccharomyces cerevisiae, and binding is inhibited by polysaccharides rich in beta1,3- or both beta1,3- and beta1,6-linked glucose. the oligosacchari ...200616371356
development and evaluation of a real-time pcr assay targeting the type iii secretion system of burkholderia pseudomallei.here we report on the development of a discriminatory real-time assay for the rapid identification of burkholderia pseudomallei isolates and the evaluation of this assay for sensitivity against related species and detection in spiked human blood samples. the assay targets a 115-base-pair region within orf2 of the b. pseudomallei type iii secretion system gene cluster and distinguishes b. pseudomallei from other microbial species. assay performance was evaluated with 224 geographically, temporall ...200616390953
plasmid-mediated high-level gentamicin resistance among enteric bacteria isolated from pet turtles in louisiana.the sale of small turtles is banned by the food and drug administration from the u.s. market due to concerns about their excretion of salmonella spp. to produce a safe pet for the export market, the louisiana pet turtle industry uses gentamicin sulfate baths (1,000 microg/ml) to eradicate salmonella spp. from turtle eggs. in 1999, we analyzed bacterial samples recovered from turtle farms and found that strains of salmonella enterica subsp. arizonae and other bacteria, such as enterobacter cloaca ...200616391058
complex regulation of arsenite oxidation in agrobacterium tumefaciens.seminal regulatory controls of microbial arsenite [as(iii)] oxidation are described in this study. transposon mutagenesis of agrobacterium tumefaciens identified genes essential for as(iii) oxidation, including those coding for a two-component signal transduction pair. the transposon interrupted a response regulator gene (referred to as aoxr), which encodes an ntrc-like protein and is immediately downstream of a gene (aoxs) encoding a protein with primary structural features found in sensor hist ...200616428412
a na+:h+ antiporter and a molybdate transporter are essential for arsenite oxidation in agrobacterium tumefaciens.transposon tn5-b22 mutagenesis was used to identify genetic determinants required for arsenite [as(iii)] oxidation in an agrobacterium tumefaciens soil isolate, strain 5a. in one mutant, the transposon interrupted modb, which codes for the permease component of a high-affinity molybdate transporter. in a second mutant, the transposon insertion occurred in mrpb, which is part of a seven-gene operon encoding an mrp-type na+:h+ antiporter complex. complementation experiments with mod and mrp operon ...200616452441
interaction of beta-1,3-glucan with its recognition protein activates hemolymph proteinase 14, an initiation enzyme of the prophenoloxidase activation system in manduca sexta.a serine proteinase pathway in insect hemolymph leads to prophenoloxidase activation, an innate immune response against pathogen infection. in the tobacco hornworm manduca sexta, recombinant hemolymph proteinase 14 precursor (pro-hp14) interacts with peptidoglycan, autoactivates, and initiates the proteinase cascade (ji, c., wang, y., guo, x., hartson, s., and jiang, h. (2004) j. biol. chem. 279, 34101-34106). here, we report the purification and characterization of pro-hp14 from the hemolymph o ...200616461344
structure-based stabilization of an enzyme: the case of penicillin acylase from alcaligenes faecalis.the modeled structure of penicillin acylase from alcaligenes faecali (afpga) was constructed by comparative modeling with the modeller program. candidate positions that could be replaced with cysteine were estimated by scanning the modeled structure of afpga with the program modip (modeling disulfide bond in protein). the mutant q3c/p751c had a higher optimum temperature by three degrees than that of the wild type afpga. the half life of the double mutant q3c/p751c at 55 degrees c was increased ...200616472081
health considerations regarding horizontal transfer of microbial transgenes present in genetically modified crops.the potential effects of horizontal gene transfer on human health are an important item in the safety assessment of genetically modified organisms. horizontal gene transfer from genetically modified crops to gut microflora most likely occurs with transgenes of microbial origin. the characteristics of microbial transgenes other than antibiotic-resistance genes in market-approved genetically modified crops are reviewed. these characteristics include the microbial source, natural function, function ...200516489267
[isolation and characterization of a chlorpyrifos degrading bacteria and its bioremediation application in the soil].a strain dsp3 capable of utilizing chlorpyrifos as the sole carbon and energy sources was isolated. based on the results of phenotypic features, phylogenetic of 16s rdna sequence, dna (g + c) mol% and dna homology between strain dsp3 and reference strains, the strain dsp3 is identified as alcaligenes faecalis. the degradation rate of chlorpyrifos was at 98.6% (100 mg/l) in liquid culture medium within 18 days and nearly 100% (100 mg/kg) in soil within 20 days respectively. an addition of strain ...200516496701
resistance determinants of a highly arsenic-resistant strain of leptospirillum ferriphilum isolated from a commercial biooxidation tank.two sets of arsenic resistance genes were isolated from the highly arsenic-resistant leptospirillum ferriphilum fairview strain. one set is located on a transposon, tnlfars, and is related to the previously identified tnatcars from acidithiobacillus caldus isolated from the same arsenopyrite biooxidation tank as l. ferriphilum. tnlfars conferred resistance to arsenite and arsenate and was transpositionally active in escherichia coli. tnlfars and tnatcars were sufficiently different for them not ...200616517682
analysis of 525 samples to determine the usefulness of pcr amplification and sequencing of the 16s rrna gene for diagnosis of bone and joint infections.the 16s rrna gene pcr in the diagnosis of bone and joint infections has not been systematically tested. five hundred twenty-five bone and joint samples collected from 525 patients were cultured and submitted to 16s rrna gene pcr detection of bacteria in parallel. the amplicons with mixed sequences were also cloned. when discordant results were observed, culture and pcr were performed once again. bacteria were detected in 139 of 525 samples. culture and 16s rrna gene pcr yielded identical documen ...200616517890
pseudomonas aeruginosa bloodstream infections: risk factors and treatment outcome related to expression of the per-1 extended-spectrum beta-lactamase.bloodstream infection (bsi) due to pseudomonas aeruginosa (pa) has relevant clinical impact especially in relation to drug resistance determinants. the per-1 extended-spectrum beta-lactamase (esbl) is a common enzyme conferring high-level resistance to anti-pseudomonal cephalosporins. risk factors and treatment outcome of bsi episodes caused by per-1-positive pa (per-1-pa) strains were compared to those caused by esbl-negative pa isolates (esbl-n-pa).200616542460
the sulfonated osmolyte n-methyltaurine is dissimilated by alcaligenes faecalis and by paracoccus versutus with release of methylamine.selective enrichments yielded bacterial cultures able to utilize the osmolyte n-methyltaurine as sole source of carbon and energy or as sole source of fixed nitrogen for aerobic growth. strain mt1, which degraded n-methyltaurine as a sole source of carbon concomitantly with growth, was identified as a strain of alcaligenes faecalis. stoichiometric amounts of methylamine, whose identity was confirmed by matrix-assisted, laser-desorption ionization time-of-flight mass spectrometry, and of sulfate ...200616549680
enhancing enzymatic activity of penicillin g acylase by coexpressing pcm gene.penicillin g acylase (pga; e.c. 3.5.1.11) is an important enzyme which has broad applications in industries of beta-lactim antibiotics production. in this study, a promising pga gene from alcaligenes faecalis (afpga) and another pcm gene encoding protein isoaspartate methyltransferase (pimt) were constructed into pet43.1a((+)) and pet28a((+)), respectively. the recombinant plasmids petafpga and petpcm were transformed into the same host cell escherichia coli bl21 (de3). results suggested that th ...200616550378
comparison of spheroplast induction in alcaligenes faecalis by three different agents.lark, cynthia (saint louis university, st. louis, mo.) and robert schichtel. comparison of spheroplast induction in alcaligenes faecalis by three different agents. j. bacteriol. 84:1241-1244. 1962.-alcaligenes faecalis strain lb was exposed to different concentrations of cycloserine, d-methionine, or penicillin. the time course of spheroplast induction by these agents was measured as a function of their concentration. the results were consistent with models in which cycloserine reversibly inhibi ...196216561981
selection and identification of bacteria isolated from waste crude oil with polycyclic aromatic hydrocarbons removal capacities.fifteen bacterial strains isolated from solid waste oil samples were selected due to their capacity of growing in the presence of hydrocarbons. the isolates were identified by pcr of the 16s rdna gene using fd1 and rd1 primers. the majority of the strains belonged to genera bacillus, bacillus pumilus (eight strains) and bacillus subtilis (two strains). besides, three strains were identified as micrococcus luteus, one as alcaligenes faecalis and one strain as enterobacter sp. growth of the above- ...200616564960
a rearranging ligand enables allosteric control of catalytic activity in copper-containing nitrite reductase.in cu-containing nitrite reductase from alcaligenes faecalis s-6 the axial methionine ligand of the type-1 site was replaced (m150g) to make the copper ion accessible to external ligands that might affect the enzyme's catalytic activity. the type-1 site optical spectrum of m150g (a(460)/a(600)=0.71) differs significantly from that of the native nitrite reductase (a(460)/a(600)=1.3). the midpoint potential of the type-1 site of nitrite reductase m150g (e(m)=312(+/-5)mv versus hydrogen) is higher ...200616574144
castellaniella gen. nov., to accommodate the phylogenetic lineage of alcaligenes defragrans, and proposal of castellaniella defragrans gen. nov., comb. nov. and castellaniella denitrificans sp. nov.comparative 16s rrna gene sequence analysis indicates that two distinct sublineages exist within the genus alcaligenes: the alcaligenes faecalis lineage, comprising alcaligenes aquatilis and a. faecalis (with the three subspecies a. faecalis subsp. faecalis, a. faecalis subsp. parafaecalis and a. faecalis subsp. phenolicus), and the alcaligenes defragrans lineage, comprising a. defragrans. this phylogenetic discrimination is supported by phenotypic and chemotaxonomic differences. it is proposed ...200616585701
16s rrna gene sequencing versus the api 20 ne system and the vitek 2 id-gnb card for identification of nonfermenting gram-negative bacteria in the clinical laboratory.over a period of 26 months, we have evaluated in a prospective fashion the use of 16s rrna gene sequencing as a means of identifying clinically relevant isolates of nonfermenting gram-negative bacilli (non-pseudomonas aeruginosa) in the microbiology laboratory. the study was designed to compare phenotypic with molecular identification. results of molecular analyses were compared with two commercially available identification systems (api 20 ne, vitek 2 fluorescent card; biomérieux, marcy l'etoil ...200616597863
cloning and sequencing of the ompa gene of enterobacter sakazakii and development of an ompa-targeted pcr for rapid detection of enterobacter sakazakii in infant formula.enterobacter sakazakii is an emerging, infant formula-borne pathogen that causes severe meningitis, meningoencephalitis, sepsis, and necrotizing enterocolitis in neonates and infants, with a high fatality rate. traditional detection methods take up to 7 days to identify e. sakazakii. the outer membrane protein a gene (ompa), along with its flanking sequences from e. sakazakii (atcc 51329), was cloned in the pgem-t easy vector and sequenced. comparison of the nucleotide and deduced amino acid seq ...200616597955
quantitative real-time legionella pcr for environmental water samples: data interpretation.quantitative legionella pcrs targeting the 16s rrna gene (specific for the genus legionella) and the mip gene (specific for the species legionella pneumophila) were applied to a total of 223 hot water system samples (131 in one laboratory and 92 in another laboratory) and 37 cooling tower samples (all in the same laboratory). the pcr results were compared with those of conventional culture. 16s rrna gene pcr results were nonquantifiable for 2.8% of cooling tower samples and up to 39.1% of hot wa ...200616597985
rapid detection of brucella spp. in blood cultures by fluorescence in situ hybridization.brucellosis is a severe systemic disease in humans. we describe a new 16s rrna-based fluorescence in situ hybridization assay that facilitates rapid and specific detection of all human pathogenic species of brucella and that can be applied directly to positive blood cultures.200616672413
stereoselective nitrile hydrolysis by immobilized whole-cell biocatalyst.the present work attempts to deal with the stability and reusability aspect of nitrilase from alcaligenes faecalis for the production of (r)-(-)-mandelic acid. four entrapment matrixes were screened to search for a suitable support, and alginate was found to have significant process advantages over its other counterparts. thermodynamic analysis allowed us to account for decreased enantioselectivity (e) as a result of immobilization. the system was also characterized based on the thiele modulus ( ...200616677036
characterization of a novel long-chain acyl-coa thioesterase from alcaligenes faecalis.a novel long-chain acyl-coa thioesterase from alcaligenes faecalis has been isolated and characterized. the protein was extracted from the cells with 1 m nacl, which required 1.5-fold, single-step purification to yield near-homogeneous preparations. in solution, the protein exists as homomeric aggregates, of mean diameter 21.6 nm, consisting of 22-kda subunits. ms/ms data for peptides obtained by trypsin digestion of the thiosterase did not match any peptide from escherichia coli thioesterases o ...200616704412
mutant afm 2 of alcaligenes faecalis for phenol biodegradation using he-ne laser irradiation.he-ne laser technology was utilized in this study to investigate the response of alcaligenes faecalis to laser stimulation. the irradiation experiments were conducted by the adjustment of the output power from 5 to 25 mw and the exposure time from 5 to 25 min. the results showed that the survival rate changed regularly with the variety of irradiation dose, and high positive mutation frequency was determined by both the energy density and the output power. the mutant strain afm 2 was obtained. ph ...200616730779
tetrathiobacter mimigardefordensis sp. nov., isolated from compost, a betaproteobacterium capable of utilizing the organic disulfide 3,3'-dithiodipropionic acid.in this study, a novel betaproteobacterium, strain dpn7(t), was isolated under mesophilic conditions from compost because of its capacity to utilize the organic disulfide 3,3'-dithiodipropionic acid. analysis of the 16s rrna gene sequence of strain dpn7(t) revealed 98.5 % similarity to that of tetrathiobacter kashmirensis lmg 22695(t). values for sequence similarity to members of the genera alcaligenes, castellaniella and taylorella, the nearest neighbours of the genus tetrathiobacter, were abou ...200616738107
studies on bacterial phosphatases: the phosphatases of aerobacter aerogenes, alcaligenes faecalis and bacillus subtilis. 193816746654
evaluation of the duopath legionella lateral flow assay for identification of legionella pneumophila and legionella species culture isolates.duopath legionella (merck kgaa, darmstadt, germany) is a new immunochromatographic assay for the simultaneous identification of cultured l. pneumophila and legionella species other than l. pneumophila. in tests of 89 l. pneumophila strains and 87 legionella strains other than l. pneumophila representing 41 different species, duopath and a widely used latex agglutination assay detected l. pneumophila with 100% and 98% accuracy, respectively, whereas the percentages differed significantly for othe ...200616751575
biology of pseudomonas stutzeri.pseudomonas stutzeri is a nonfluorescent denitrifying bacterium widely distributed in the environment, and it has also been isolated as an opportunistic pathogen from humans. over the past 15 years, much progress has been made in elucidating the taxonomy of this diverse taxonomical group, demonstrating the clonality of its populations. the species has received much attention because of its particular metabolic properties: it has been proposed as a model organism for denitrification studies; many ...200616760312
uronate isomerase: a nonhydrolytic member of the amidohydrolase superfamily with an ambivalent requirement for a divalent metal ion.uronate isomerase, a member of the amidohydrolase superfamily, catalyzes the isomerization of d-glucuronate and d-fructuronate. during the interconversion of substrate and product the hydrogen at c2 of d-glucuronate is transferred to the pro-r position at c1 of the product, d-fructuronate. the exchange of the transferred hydrogen with solvent deuterium occurs at a rate that is 4 orders of magnitude slower than the interconversion of substrate and product. the enzyme catalyzes the elimination of ...200616768441
surface display of proteins by gram-negative bacterial autotransporters.expressing proteins of interest as fusions to proteins of the bacterial envelope is a powerful technique with many biotechnological and medical applications. autotransporters have recently emerged as a good tool for bacterial surface display. these proteins are composed of an n-terminal signal peptide, followed by a passenger domain and a translocator domain that mediates the outer membrane translocation of the passenger. the natural passenger domain of autotransporters can be replaced by hetero ...200616787545
immobilization of permeabilized whole cell penicillin g acylase from alcaligenes faecalis using pore matrix crosslinked with glutaraldehyde.the activity of penicillin g acylase from alcaligenes faecalis increased 7.5-fold when cells were permeabilized with 0.3% (w/v) ctab. the treated cells were entrapped by polyvinyl alcohol crosslinked with boric acid, and crosslinked with 2% (v/v) glutaraldehyde to increase the stability. the conversion yield of penicillin g to 6-aminopenicillanic acid was 75% by immobilized system in batch reaction. no activity was lost after 15 cycles and about 65% enzyme activity was retained at the end of the ...200616799762
evaluation of pyrrolidonyl arylamidase for the identification of nonfermenting gram-negative rods.to evaluate the activity of pyrrolidonyl arylamidase (pyr) for the differentiation and identification of nonfermenting gram negative rods (nfgnr), 293 isolates were tested. a 24 h culture of each test organism was prepared. from this a 108-109 cfu/ml suspension was added to 0.25 ml of sterile physiologic solution. a pyr disk was then added and the test was incubated for 30 minutes at 35-37 degrees c, at environmental atmosphere. reading was done by adding 1 drop of cinnamaldehyde reagent. strain ...200716822636
quantitative detection of the nosz gene, encoding nitrous oxide reductase, and comparison of the abundances of 16s rrna, narg, nirk, and nosz genes in soils.nitrous oxide (n2o) is an important greenhouse gas in the troposphere controlling ozone concentration in the stratosphere through nitric oxide production. in order to quantify bacteria capable of n2o reduction, we developed a sybr green quantitative real-time pcr assay targeting the nosz gene encoding the catalytic subunit of the nitrous oxide reductase. two independent sets of nosz primers flanking the nosz fragment previously used in diversity studies were designed and tested (k. kloos, a. mer ...200616885263
metabolic profiles and genetic diversity of denitrifying communities in activated sludge after addition of methanol or ethanol.external carbon sources can enhance denitrification rates and thus improve nitrogen removal in wastewater treatment plants. the effects of adding methanol and ethanol on the genetic and metabolic diversity of denitrifying communities in activated sludge were compared using a pilot-scale plant with two parallel lines. a full-scale plant receiving the same municipal wastewater, but without external carbon source addition, was the reference. metabolic profiles obtained from potential denitrificatio ...200616885297
piggery wastewater treatment using alcaligenes faecalis strain no. 4 with heterotrophic nitrification and aerobic denitrification.alcaligenes faecalis strain no. 4, which has heterotrophic nitrification and aerobic denitrification abilities, was used to treat actual piggery wastewater containing high-strength ammonium under aerobic conditions. in a continuous experiment using a solids-free wastewater (sfw) mixed with feces, almost all of the 2000 nh4+ -n mg/l and 12,000 cod mg/l in the wastewater was removed and the ammonium removal rate was approximately 30 mg-n/l/h, which was 5-10 times higher than the rates achieved by ...200616893560
effect of ph, temperature and substrate on n2o, no and co2 production by alcaligenes faecalis p.to study the effect of ph, temperature and substrate on the magnitude of n(2)o and no production by heterotrophic nitrifiers.200616907816
fluorescence in situ hybridization for rapid identification of achromobacter xylosoxidans and alcaligenes faecalis recovered from cystic fibrosis patients.achromobacter xylosoxidans is frequently isolated from the respiratory secretions of cystic fibrosis (cf) patients, but identification with biochemical tests is unreliable. we describe fluorescence in situ hybridization assays for the rapid identification of achromobacter xylosoxidans and alcaligenes faecalis. both assays showed high sensitivities and high specificities with a collection of 155 nonfermenters from cf patients.200616954289
quantitative determination of free-dna uptake in river bacteria at the single-cell level by in situ rolling-circle amplification.detection of plasmid dna uptake in river bacteria at the single-cell level was carried out by rolling-circle amplification (rca). uptake of a plasmid containing the green fluorescent protein gene (gfp) by indigenous bacteria from two rivers in osaka, japan, was monitored for 506 h using this in situ gene amplification technique with optimized cell permeabilization conditions. plasmid uptake determined by in situ rca was compared to direct counts of cells expressing gfp under fluorescence microsc ...200616957252
effects of mutations in the substrate-binding domain of poly[(r)-3-hydroxybutyrate] (phb) depolymerase from ralstonia pickettii t1 on phb degradation.poly[(r)-3-hydroxybutyrate] (phb) depolymerase from ralstonia pickettii t1 (phaz(rpit1)) adsorbs to denatured phb (dphb) via its substrate-binding domain (sbd) to enhance dphb degradation. to evaluate the amino acid residues participating in dphb adsorption, phaz(rpit1) was subjected to a high-throughput screening system consisting of pcr-mediated random mutagenesis targeted to the sbd gene and a plate assay to estimate the effects of mutations in the sbd on dphb degradation by phaz(rpit1). gene ...200616963553
probing the unusual oxidation/reduction behavior of paracoccus pantotrophus cytochrome cd1 nitrite reductase by replacing a switchable methionine heme iron ligand with histidine.a m106h variant, where m106 is a c-type heme iron axial ligand, of cytochrome cd(1) nitrite reductase is an inactive protein in vivo. expression of the holoprotein in paracoccus pantotrophus required generation of nitric oxide during cell growth through simultaneous expression of an exogenous copper nitrite reductase from alcaligenes faecalis. in the absence of the latter protein, only a semi-apo form of m106h cytochrome cd(1) was formed. thus it was demonstrated that expression of the chromosom ...200616964982
the efficacy of postenucleation saline wash and the effect of different antimicrobial agents on microbial contamination of donor eyes.to evaluate the efficacy of postenucleation saline wash and the effect of different antimicrobial agents on the microbial contamination of donor eyes.200616974130
two-center collaborative evaluation of performance of the bd phoenix automated microbiology system for identification and antimicrobial susceptibility testing of gram-negative bacteria.the performance of the bd phoenix automated microbiology system (bd diagnostic systems, sparks, md) was assessed for identification (id) and antimicrobial susceptibility testing (ast) of the majority of clinically encountered bacterial isolates in a european collaborative two-center trial. a total of 494 bacterial isolates including various species of the enterobacteriaceae and 110 nonfermentative gram-negative bacteria were investigated: of these, 385 were single patient isolates, and 109 were ...200617005752
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