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bacterial cell factories for recombinant protein production; expanding the catalogue. 201324245806
mechanism and catalytic diversity of rieske non-heme iron-dependent oxygenases.rieske non-heme iron-dependent oxygenases are important enzymes that catalyze a wide variety of reactions in the biodegradation of xenobiotics and the biosynthesis of bioactive natural products. in this perspective article, we summarize recent efforts to elucidate the catalytic mechanisms of rieske oxygenases and highlight the diverse range of reactions now known to be catalyzed by such enzymes.201324244885
identification of new residues involved in intramolecular signal transmission in a prokaryotic transcriptional repressor.ttgv is a member of the iclr family of transcriptional regulators. this regulator controls its own expression and that of the ttgghi operon, which encodes an rnd efflux pump. ttgv has two domains: a gaf-like domain harboring the effector-binding pocket and a helix-turn-helix (hth) dna-binding domain, which are linked by a long extended helix. when ttgv is bound to dna, a kink at residue 86 in the extended helix gives rise to 2 helices. ttgv contacts dna mainly through a canonical recognition hel ...201424244009
actinobacterial acyl coenzyme a synthetases involved in steroid side-chain catabolism.bacterial steroid catabolism is an important component of the global carbon cycle and has applications in drug synthesis. pathways for this catabolism involve multiple acyl coenzyme a (coa) synthetases, which activate alkanoate substituents for β-oxidation. the functions of these synthetases are poorly understood. we enzymatically characterized four distinct acyl-coa synthetases from the cholate catabolic pathway of rhodococcus jostii rha1 and the cholesterol catabolic pathway of mycobacterium t ...201424244004
iron(ii)-dependent dioxygenase and n-formylamide deformylase catalyze the reactions from 5-hydroxy-2-pyridone to maleamate.5-hydroxy-2-pyridone (2,5-dhp) is a central metabolic intermediate in catabolism of many pyridine derivatives, and has been suggested as a potential carcinogen. 2,5-dhp is frequently transformed to n-formylmaleamic acid (nfm) by a 2,5-dhp dioxygenase. three hypotheses were formerly discussed for conversion of 2,5-dhp to maleamate. based on enzymatic reactions of dioxygenase (hpo) and n-formylamide deformylase (nfo), we demonstrated that the dioxygenase does not catalyze the hydrolysis of nfm but ...201324241081
spalf4: a newly identified anti-lipopolysaccharide factor from the mud crab scylla paramamosain with broad spectrum antimicrobial activity.anti-lipopolysaccharide factors (alfs) are antimicrobial peptides with binding and neutralizing activities to lipopolysaccharide (lps) in crustaceans. this study identified and characterized a novel alf homolog (spalf4) from the mud crab scylla paramamosain. the complete cdna of spalf4 had 756 bp with a 381 bp open reading frame encoding a protein with 126 aa. the deduced protein contained a signal peptide and a lps-binding domain. spalf4 shared the highest identity with ptalf5 at amino acid lev ...201424239582
functional characterization of an ornithine cyclodeaminase-like protein of arabidopsis thaliana.in plants, proline synthesis occurs by two enzymatic steps starting from glutamate as a precursor. some bacteria, including bacteria such as agrobacterium rhizogenes have an ornithine cyclodeaminase (ocd) which can synthesize proline in a single step by deamination of ornithine. in a. rhizogenes, ocd is one of the genes transferred to the plant genome during the transformation process and plants expressing a. rhizogenes ocd have developmental phenotypes. one nuclear encoded gene of arabidopsis t ...201324237637
enhanced cadmium resistance and accumulation in pseudomonas putida kt2440 expressing the phytochelatin synthase gene of schizosaccharomyces pombe.phytochelatins (pcs) are cysteine-rich peptides with high binding affinity for toxic metals. expressing the pc synthase gene (pcs) in plant growth-promoting bacteria may enhance its metal resistance and accumulation, consequently increasing phytoremediation efficiency in heavy metal pollution. in this study, pcs from schizosaccharomyces pombe was cloned and expressed in pseudomonas putida kt2440, which was confirmed by real-time rt-pcr through an increase in sppcs mrna expression level when indu ...201424236847
diversity and evolution of bacterial twin arginine translocase protein, tatc, reveals a protein secretion system that is evolving to fit its environmental niche.the twin-arginine translocation (tat) protein export system enables the transport of fully folded proteins across a membrane. this system is composed of two integral membrane proteins belonging to tata and tatc protein families and in some systems a third component, tatb, a homolog of tata. tatc participates in substrate protein recognition through its interaction with a twin arginine leader peptide sequence.201324236045
so depression is an inflammatory disease, but where does the inflammation come from?we now know that depression is associated with a chronic, low-grade inflammatory response and activation of cell-mediated immunity, as well as activation of the compensatory anti-inflammatory reflex system. it is similarly accompanied by increased oxidative and nitrosative stress (o&ns), which contribute to neuroprogression in the disorder. the obvious question this poses is 'what is the source of this chronic low-grade inflammation?'201324228900
the metacyc database of metabolic pathways and enzymes and the biocyc collection of pathway/genome databases.the metacyc database (metacyc.org) is a comprehensive and freely accessible database describing metabolic pathways and enzymes from all domains of life. metacyc pathways are experimentally determined, mostly small-molecule metabolic pathways and are curated from the primary scientific literature. metacyc contains >2100 pathways derived from >37,000 publications, and is the largest curated collection of metabolic pathways currently available. biocyc (biocyc.org) is a collection of >3000 organism- ...201324225315
the metacyc database of metabolic pathways and enzymes and the biocyc collection of pathway/genome databases.the metacyc database (metacyc.org) is a comprehensive and freely accessible database describing metabolic pathways and enzymes from all domains of life. metacyc pathways are experimentally determined, mostly small-molecule metabolic pathways and are curated from the primary scientific literature. metacyc contains >2100 pathways derived from >37,000 publications, and is the largest curated collection of metabolic pathways currently available. biocyc (biocyc.org) is a collection of >3000 organism- ...201324225315
water-limiting conditions alter the structure and biofilm-forming ability of bacterial multispecies communities in the alfalfa rhizosphere.biofilms are microbial communities that adhere to biotic or abiotic surfaces and are enclosed in a protective matrix of extracellular compounds. an important advantage of the biofilm lifestyle for soil bacteria (rhizobacteria) is protection against water deprivation (desiccation or osmotic effect). the rhizosphere is a crucial microhabitat for ecological, interactive, and agricultural production processes. the composition and functions of bacterial biofilms in soil microniches are poorly underst ...201324223979
microbial environment affects innate immunity in two closely related earthworm species eisenia andrei and eisenia fetida.survival of earthworms in the environment depends on their ability to recognize and eliminate potential pathogens. this work is aimed to compare the innate defense mechanisms of two closely related earthworm species, eisenia andrei and eisenia fetida, that inhabit substantially different ecological niches. while e. andrei lives in a compost and manure, e. fetida can be found in the litter layer in forests. therefore, the influence of environment-specific microbiota on the immune response of both ...201324223917
structural and enzymatic characterization of the phosphotriesterase ophc2 from pseudomonas pseudoalcaligenes.organophosphates (ops) are neurotoxic compounds for which current methods of elimination are unsatisfactory; thus bio-remediation is considered as a promising alternative. here we provide the structural and enzymatic characterization of the recently identified enzyme isolated from pseudomonas pseudoalcaligenes dubbed ophc2. ophc2 belongs to the metallo-β-lactamase superfamily and exhibits an unusual thermal resistance and some op degrading abilities.201324223749
comparison of biofilm and attachment mechanisms of a phytopathological and clinical isolate of klebsiella pneumoniae subsp. pneumoniae.some bacterial species can colonize humans and plants. it is almost impossible to prevent the contact of clinically pathogenic bacteria with food crops, and if they can persist there, they can reenter the human food chain and cause disease. on the leaf surface, microorganisms are exposed to a number of stress factors. it is unclear how they survive in such different environments. by increasing adhesion to diverse substrates, minimizing environmental differences, and providing protection against ...201324222755
influence of root exudates on the bacterial degradation of chlorobenzoic acids.degradation of chlorobenzoic acids (e.g., products of microbial degradation of pcb) by strains of microorganisms isolated from pcb contaminated soils was assessed. from seven bulk-soil isolates two strains unique in ability to degrade a wider range of chlorobenzoic acids than others were selected, individually and even in a complex mixture of 11 different chlorobenzoic acids. such a feature is lacking in most tested degraders. to investigate the influence of vegetation on chlorobenzoic acids deg ...201324222753
episodes of horizontal gene-transfer and gene-fusion led to co-existence of different metal-ion specific glyoxalase i.glyoxalase pathway plays an important role in stress adaptation and many clinical disorders. the first enzyme of this pathway, glyoxalase i (glxi), uses methylglyoxal as a substrate and requires either ni(ii)/co(ii) or zn(ii) for activity. here we have investigated the origin of different metal ion specificities of glxi and subsequent pattern of inheritance during evolution. our results suggest a primitive origin of single-domain ni dependent glxi [ni-glxi]. this subsequently evolved into zn act ...201324220130
development of a broad-host synthetic biology toolbox for ralstonia eutropha and its application to engineering hydrocarbon biofuel production.the chemoautotrophic bacterium ralstonia eutropha can utilize h2/co2 for growth under aerobic conditions. while this microbial host has great potential to be engineered to produce desired compounds (beyond polyhydroxybutyrate) directly from co2, little work has been done to develop genetic part libraries to enable such endeavors.201324219429
biochemical and biophysical characterization of the two isoforms of cbb3-type cytochrome c oxidase from pseudomonas stutzeri.the cbb3-type cytochrome c oxidases (cbb3-ccos) are members of the heme-copper oxidase superfamily that couple the reduction of oxygen to translocation of protons across the membrane. the cbb3-ccos are present only in bacteria and play a primary role in microaerobic respiration, being essential for nitrogen-fixing endosymbionts and for some human pathogens. as frequently observed in pseudomonads, pseudomonas stutzeri contains two independent ccono(q)p operons encoding the two cbb3 isoforms, cbb3 ...201424214947
a plant selectable marker gene based on the detoxification of the herbicide dalapon.a gene from pseudomonas putida coding for a dehalogenase capable of degrading 2,2 dichloropropionic acid (2,2dcpa), the active ingredient of the herbicide dalapon, has been isolated and characterised. in plant transformation experiments the gene was shown to confer resistance to 2,2dcpa at a tissue culture level where 2,2dcpa could be used to select for transformants. at the whole plant level, transformed plants showed resistance to 2,2dcpa at concentrations up to 5 times the recommended dose ra ...199224213366
pham is the physiological activator of poly(3-hydroxybutyrate) (phb) synthase (phac1) in ralstonia eutropha.poly(3-hydroxybutyrate) (phb) synthase (phac1) is the key enzyme of phb synthesis in ralstonia eutropha and other phb-accumulating bacteria and catalyzes the polymerization of 3-hydroxybutyryl-coa to phb. activity assays of r. eutropha phb synthase are characterized by the presence of lag phases and by low specific activity. it is assumed that the lag phase is caused by the time necessary to convert the inactive phac1 monomer into the active dimeric form by an unknown priming process. the lag ph ...201424212577
development and characterization of microbial biosensors for evaluating low biochemical oxygen demand in rivers.five microorganisms were used to construct a biosensor for the evaluation of low biochemical oxygen demand (bod) in rivers. characterization and comparison of bod biosensors were performed using two standard solutions: glucose and glutamic acid (gga) and artificial wastewater (aww). pseudomonas putida sg10 demonstrated the best response when using aww. trichosporon cutaneum ifo10466, however, had an extremely poor response. when evaluating the biosensor response to each component of aww, all of ...201324209354
periplasmic glucose-binding protein from pseudomonas putida csv86--identification of the glucose-binding pocket by homology-model-guided site-specific mutagenesis.glucose transport in pseudomonas putida csv86 is mediated via a periplasmic glucose-binding protein (gbp)-dependent putative glucose abc transporter. here we describe a homology model and functional characterization of gbp from csv86 (ppgbp). a whole-cell [(14)c]-glucose uptake study revealed that glucose is transported by the high-affinity intracellular phosphorylative pathway. ppgbp was cloned, over-expressed in escherichia coli and purified to apparent homogeneity. the purified ppgbps from bo ...201424206004
mechanisms of copper homeostasis in bacteria.copper is an important micronutrient required as a redox co-factor in the catalytic centers of enzymes. however, free copper is a potential hazard because of its high chemical reactivity. consequently, organisms exert a tight control on cu(+) transport (entry-exit) and traffic through different compartments, ensuring the homeostasis required for cuproprotein synthesis and prevention of toxic effects. recent studies based on biochemical, bioinformatics, and metalloproteomics approaches, reveal a ...201324205499
novel tools to analyze the function of salmonella effectors show that svpb ectopic expression induces cell cycle arrest in tumor cells.in order to further characterize its role in pathogenesis and to establish whether its overproduction can lead to eukaryotic tumor cell death, salmonella strains able to express its virulence factor spvb (an adp-ribosyl transferase enzyme) in a salicylate-inducible way have been constructed and analyzed in different eukaryotic tumor cell lines. to do so, the bacterial strains bearing the expression system have been constructed in a ∆purd background, which allows control of bacterial proliferatio ...201324205236
biochemical characterization of a first fungal esterase from rhizomucor miehei showing high efficiency of ester synthesis.esterases with excellent merits suitable for commercial use in ester production field are still insufficient. the aim of this research is to advance our understanding by seeking for more unusual esterases and revealing their characterizations for ester synthesis.201324204998
systematic unraveling of the unsolved pathway of nicotine degradation in pseudomonas.microorganisms such as pseudomonas putida play important roles in the mineralization of organic wastes and toxic compounds. to comprehensively and accurately elucidate key processes of nicotine degradation in pseudomonas putida, we measured differential protein abundance levels with ms-based spectral counting in p. putida s16 grown on nicotine or glycerol, a non-repressive carbon source. in silico analyses highlighted significant clustering of proteins involved in a functional pathway in nicotin ...201324204321
novel methods for analysing bacterial tracks reveal persistence in rhodobacter sphaeroides.tracking bacteria using video microscopy is a powerful experimental approach to probe their motile behaviour. the trajectories obtained contain much information relating to the complex patterns of bacterial motility. however, methods for the quantitative analysis of such data are limited. most swimming bacteria move in approximately straight lines, interspersed with random reorientation phases. it is therefore necessary to segment observed tracks into swimming and reorientation phases to extract ...201324204227
thiosulfate oxidation by obligately heterotrophic bacteria.thiosulfate was oxidized stoichiometrically to tetrathionate during growth on glucose byklebsiella aerogenes, bacillus globigii, b. megaterium, pseudomonas putida, two strains each ofp. fluorescens andp. aeruginosa, and anaeromonas sp. a gram-negative, rod-shaped soil isolate, pseudomonad hw, converted thiosulfate to tetrathionate during growth on acetate. none of the organisms could use thiosulfate as sole energy source. the quantitative recovery of all the thiosulfate supplied to heterotrophic ...198824202996
the flavin reductase msue is a novel nitroreductase that can efficiently activate two promising next-generation prodrugs for gene-directed enzyme prodrug therapy.bacterial nitroreductase enzymes that can efficiently catalyse the oxygen-independent reduction of prodrugs originally developed to target tumour hypoxia offer great potential for expanding the therapeutic range of these molecules to aerobic tumour regions, via the emerging cancer strategy of gene-directed enzyme prodrug therapy (gdept). two promising hypoxia prodrugs for gdept are the dinitrobenzamide mustard pr-104a, and the nitrochloromethylbenzindoline prodrug nitro-cbi-dei. we describe here ...201324202330
partial bioaugmentation to remove 3-chloroaniline slows bacterial species turnover rate in bioreactors.bioaugmentation is a potentially powerful tool to direct community structure and metabolic capacities in bioreactors. yet the outcome of bioaugmentation studies is usually unpredictable and effects on microbial community dynamics are poorly understood. we asked the question whether bioaugmentation could prevent a diversity shift induced by a model toxin, 3-chloroaniline (3-ca), regardless of whether 3-ca was degraded. four replicate membrane bioreactors (mbrs) operating in parallel were amended ...201324200008
the microbiology of olive mill wastes.olive mill wastes (omws) are high-strength organic effluents, which upon disposal can degrade soil and water quality, negatively affecting aquatic and terrestrial ecosystems. the main purpose of this review paper is to provide an up-to-date knowledge concerning the microbial communities identified over the past 20 years in olive mill wastes using both culture-dependent and independent approaches. a database survey of 16s rrna gene sequences (585 records in total) obtained from olive mill waste e ...201324199199
long-term survival of and plasmid stability inpseudomonas andklebsiella species and appearance of nonculturable cells in agricultural drainage water.one year after introduction into agricultural drainage waterpseudomonas fluorescens r2f (rp4),pseudomonas putida cym318 (prk2501), andklebsiella aerogenes nctc418 (pbr322) could be recovered on agar media. survival of the introduced strains depended on competition with the indigenous microflora, the presence of nutrients, and the availability of air.in contrast tok. aerogenes nctc418 (pbr322), bothpseudomonas species lost their plasmids, as indicated by the consistently lower colony counts on se ...199024196361
viable but nonculturable bacteria: food safety and public health perspective.the viable but nonculturable (vbnc) state is a unique survival strategy of many bacteria in the environment in response to adverse environmental conditions. vbnc bacteria cannot be cultured on routine microbiological media, but they remain viable and retain virulence. the vbnc bacteria can be resuscitated when provided with appropriate conditions. a good number of bacteria including many human pathogens have been reported to enter the vbnc state. though there have been disputes on the existence ...201324191231
functional assembly of camphor converting two-component baeyer-villiger monooxygenases with a flavin reductase from e. coli.the major limitation in the synthetic application of two-component baeyer-villiger monooxygenases was addressed by identifying the 28-kda flavin-reductase fre from escherichia coli as a suitable supplier of reduced fmn for these enzymes. coexpression of fre with either 2,5- or 3,6-diketocamphane monooxygenase from pseudomonas putida ncimb 10007 significantly enhanced the conversion of camphor and norcamphor serving as representative ketones. with purified enzymes, full conversion was achieved, w ...201424190498
fate of pseudomonas putida after release into lake water mesocosms: different survival mechanisms in response to environmental conditions.to study the fate of pseudomonas putida dsm 3931 in an aquatic environment, cultures of the strain were released into lake water mesocosms. p. putida, bearing the tol-plasmid, was released as a representative xenobiotic-degrading microorganism. the release was carried out in mesocosms with unamended lake water and in lake water with added culture medium to compare the survival of the strain due to the influence of different organic load. as a comparison, the survival of p. putida was followed in ...199424190270
biofilms: the stronghold of legionella pneumophila.legionellosis is mostly caused by legionella pneumophila and is defined as a severe respiratory illness with a case fatality rate ranging from 5% to 80%. l. pneumophila is ubiquitous in natural and anthropogenic water systems. l. pneumophila is transmitted by inhalation of contaminated aerosols produced by a variety of devices. while l. pneumophila replicates within environmental protozoa, colonization and persistence in its natural environment are also mediated by biofilm formation and coloniza ...201324185913
the basidiomycetous yeast trichosporon may cause severe lung exacerbation in cystic fibrosis patients - clinical analysis of trichosporon positive patients in a munich cohort.the relevance of trichosporon species for cystic fibrosis (cf) patients has not yet been extensively investigated.201324180379
structure and function of the trp3 gene of saccharomyces cerevisiae: analysis of transcription, promoter sequence, and sequence coding for a glutamine amidotransferase.the structure and function of the trp3 gene of saccharomyces cerevisiae were analyzed. subcloning of an original 4.8 kb bamhi dna fragment, carrying the yeast trp3 gene, allowed for a localization of the gene on a 2.5 kb clai/bamhi fragment. transcription was found to proceed from the clai site towards the bamhi site. three major transcription start sites were determined at positions -92, -87, and -81 by s1-mapping. the synthesis of the trp3 gene is regulated by the general control, and was foun ...198424177735
crystal structure of a putative quorum sensing-regulated protein (pa3611) from the pseudomonas-specific duf4146 family.pseudomonas aeruginosa is an opportunistic pathogen commonly found in humans and other organisms and is an important cause of infection especially in patients with compromised immune defense mechanisms. the pa3611 gene of p. aeruginosa pao1 encodes a secreted protein of unknown function, which has been recently classified into a small pseudomonas-specific protein family called duf4146. as part of our effort to extend structural coverage of novel protein space and provide a structure-based functi ...201324174223
crystal structure of a putative quorum sensing-regulated protein (pa3611) from the pseudomonas-specific duf4146 family.pseudomonas aeruginosa is an opportunistic pathogen commonly found in humans and other organisms and is an important cause of infection especially in patients with compromised immune defense mechanisms. the pa3611 gene of p. aeruginosa pao1 encodes a secreted protein of unknown function, which has been recently classified into a small pseudomonas-specific protein family called duf4146. as part of our effort to extend structural coverage of novel protein space and provide a structure-based functi ...201324174223
bacterial farming by the fungus morchella crassipes.the interactions between bacteria and fungi, the main actors of the soil microbiome, remain poorly studied. here, we show that the saprotrophic and ectomycorrhizal soil fungus morchella crassipes acts as a bacterial farmer of pseudomonas putida, which serves as a model soil bacterium. farming by m. crassipes consists of bacterial dispersal, bacterial rearing with fungal exudates, as well as harvesting and translocation of bacterial carbon. the different phases were confirmed experimentally using ...201324174111
reconciling in vivo and in silico key biological parameters of pseudomonas putida kt2440 during growth on glucose under carbon-limited condition.genome scale metabolic reconstructions are developed to efficiently engineer biocatalysts and bioprocesses based on a rational approach. however, in most reconstructions, due to the lack of appropriate measurements, experimentally determined growth parameters are simply taken from literature including other organisms, which reduces the usefulness and suitability of these models. pseudomonas putida kt2440 is an outstanding biocatalyst given its versatile metabolism, its ability to generate suffic ...201324168623
two-way plant mediated interactions between root-associated microbes and insects: from ecology to mechanisms.plants are members of complex communities and function as a link between above- and below-ground organisms. associations between plants and soil-borne microbes commonly occur and have often been found beneficial for plant fitness. root-associated microbes may trigger physiological changes in the host plant that influence interactions between plants and aboveground insects at several trophic levels. aboveground, plants are under continuous attack by insect herbivores and mount multiple responses ...201324167508
comparative metabolic systems analysis of pathogenic burkholderia.burkholderia cenocepacia and burkholderia multivorans are opportunistic drug-resistant pathogens that account for the majority of burkholderia cepacia complex infections in cystic fibrosis patients and also infect other immunocompromised individuals. while they share similar genetic compositions, b. cenocepacia and b. multivorans exhibit important differences in pathogenesis. we have developed reconciled genome-scale metabolic network reconstructions of b. cenocepacia j2315 and b. multivorans at ...201424163337
a moderate toxin, grat, modulates growth rate and stress tolerance of pseudomonas putida.chromosomal toxin-antitoxin (ta) systems are widespread among free-living bacteria and are supposedly involved in stress tolerance. here, we report the first ta system identified in the soil bacterium pseudomonas putida. the system, encoded by the loci pp1586-pp1585, is conserved in pseudomonads and belongs to the higba family. the new ta pair was named grata for the growth rate-affecting ability of grat and the antidote activity of graa. the grata system shares many features common to previousl ...201424163334
medium chain length polyhydroxyalkanoate (mcl-pha) production from volatile fatty acids derived from the anaerobic digestion of grass.a two step biological process for the conversion of grass biomass to the biodegradable polymer medium chain length polyhydroxyalkanoate (mcl-pha) was achieved through the use of anaerobic and aerobic microbial processes. anaerobic digestion (mixed culture) of ensiled grass was achieved with a recirculated leach bed bioreactor resulting in the production of a leachate, containing 15.3 g/l of volatile fatty acids (vfas) ranging from acetic to valeric acid with butyric acid predominating (12.8 g/l) ...201424162086
denitrification of industrial wastewater: influence of glycerol addition on metabolic activity and community shifts in a microbial consortium.the wastewater originating from explosives manufacturing plants are characterized by a high concentration of nitrates (3200mgnl(-1)), sulfates (1470mgl(-1)) and low ph (1.5) as well as the presence of organic compounds, such as nitroglycerin (1.9mgl(-1)) and nitroglycol (4.8mgl(-1)). the application of glycerol (c/n=3) at such a high concentration enabled complete removal of nitrates and did not cause the anaerobic glycerol metabolic pathway of the dnc4 consortium to activate, as confirmed by th ...201324161581
properties of catechol 1,2-dioxygenase in the cell free extract and immobilized extract of mycobacterium fortuitum.polycyclic aromatic hydrocarbons (pah) are carcinogenic compounds which contaminate water and soil, and the enzymes can be used for bioremediation of these environments. this study aimed to evaluate some environmental conditions that affect the production and activity of the catechol 1,2-dioxygenase (c12o) by mycobacterium fortuitum in the cell free and immobilized extract in sodium alginate. the bacterium was grown in mineral medium and lb broth containing 250 mg l(-1) of anthracene (pah). the ...201324159319
patient-specific analysis of periodontal and peri-implant microbiomes.periodontally involved teeth have been implicated as 'microbial reservoirs' in the etiology of peri-implant diseases. therefore, the purpose of this investigation was to use a deep-sequencing approach to identify the degree of congruence between adjacent peri-implant and periodontal microbiomes in states of health and disease. subgingival and peri-implant biofilm samples were collected from 81 partially edentulous individuals with periodontal and peri-implant health and disease. bacterial dna wa ...201324158341
fad c(4a)-hydroxide stabilized in a naturally fused styrene monooxygenase.stya2b represents a new class of styrene monooxygenases that integrates flavin-reductase and styrene-epoxidase activities into a single polypeptide. this naturally-occurring fusion protein offers new avenues for studying and engineering biotechnologically relevant enantioselective biochemical epoxidation reactions. stopped-flow kinetic studies of stya2b reported here identify reaction intermediates similar to those reported for the separate reductase and epoxidase components of related two-compo ...201324157359
comparative proteomic analysis reveals mechanistic insights into pseudomonas putida f1 growth on benzoate and citrate.pseudomonas species are capable to proliferate under diverse environmental conditions and thus have a significant bioremediation potential. to enhance our understanding of their metabolic versatility, this study explores the changes in the proteome and physiology of pseudomonas putida f1 resulting from its growth on benzoate, a moderate toxic compound that can be catabolized, and citrate, a carbon source that is assimilated through central metabolic pathways. a series of repetitive batch cultiva ...201324156539
uncovering the determinants of a highly perturbed tyrosine pka in the active site of ketosteroid isomerase.within the idiosyncratic enzyme active-site environment, side chain and ligand pka values can be profoundly perturbed relative to their values in aqueous solution. whereas structural inspection of systems has often attributed perturbed pka values to dominant contributions from placement near charged groups or within hydrophobic pockets, tyr57 of a pseudomonas putida ketosteroid isomerase (ksi) mutant, suggested to have a pka perturbed by nearly 4 units to 6.3, is situated within a solvent-expose ...201324151972
improved production of medium-chain-length polyhydroxyalkanoates in glucose-based fed-batch cultivations of metabolically engineered pseudomonas putida strains.one of the major challenges in metabolic engineering for enhanced synthesis of value-added chemicals is to design and develop new strains that can be translated into well-controlled fermentation processes using bioreactors. the aim of this study was to assess the influence of various fed-batch strategies in the performance of metabolically engineered pseudomonas putida strains, δgcd and δgcd-pgl, for improving production of medium-chain-length polyhydroxyalkanoates (mcl-phas) using glucose as th ...201424150495
the metabolic cost of flagellar motion in pseudomonas putida kt2440.although the flagellar machinery of environmental bacteria endows cells with a phenomenal survival device, it also consumes much of the metabolic currency necessary for fuelling such a vigorous nano-motor. the physiological cost of flagella-related functions of the soil bacterium pseudomonas putida kt2440 was examined and quantified through the deletion of a ≈ 70 kb dna segment of the genome (≈ 1.1%), which includes relevant structural and regulatory genes in this micro-organism. the resulting s ...201424148021
manganese (mn) oxidation increases intracellular mn in pseudomonas putida gb-1.bacterial manganese (mn) oxidation plays an important role in the global biogeochemical cycling of mn and other compounds, and the diversity and prevalence of mn oxidizers have been well established. despite many hypotheses of why these bacteria may oxidize mn, the physiological reasons remain elusive. intracellular mn levels were determined for pseudomonas putida gb-1 grown in the presence or absence of mn by inductively coupled plasma mass spectrometry (icp-ms). mn oxidizing wild type p. putid ...201324147089
reprogramming of fatty acid and oxylipin synthesis in rhizobacteria-induced systemic resistance in tomato.the rhizobacterium pseudomonas putida btp1 stimulates induced systemic resistance (isr) in tomato. a previous work showed that the resistance is associated in leaves with the induction of the first enzyme of the oxylipin pathway, the lipoxygenase (lox), leading to a faster accumulation of its product, the free 13-hydroperoxy octadecatrienoic acid (13-hpot), 2 days after botrytis cinerea inoculation. in the present study, we further investigated the stimulation of the oxylipin pathway: metabolite ...201424146221
identification of mur34 as the novel negative regulator responsible for the biosynthesis of muraymycin in streptomyces sp. nrrl30471.muraymycin, a potent translocase i (mray) inhibitor, is produced by streptomyces sp. nrrl30471. the muraymycin gene cluster (mur) was recently cloned, and bioinformatic analysis of mur34 revealed its encoding product exhibits high homology to a large family of proteins, including kani and raci in individual biosynthetic pathway of kanamycin and ribostamycin. however, the precise role of these proteins remains unknown.201324143177
moraxella catarrhalis expresses a cardiolipin synthase that impacts adherence to human epithelial cells.the major phospholipid constituents of moraxella catarrhalis membranes are phosphatidylglycerol, phosphatidylethanolamine, and cardiolipin (cl). however, very little is known regarding the synthesis and function of these phospholipids in m. catarrhalis. in this study, we discovered that m. catarrhalis expresses a cardiolipin synthase (cls), termed mcls, that is responsible for the synthesis of cl within the bacterium. the nucleotide sequence of mcls is highly conserved among m. catarrhalis isola ...201424142255
microbiomes of unreactive and pathologically altered ileocecal lymph nodes of slaughter pigs.microbe-laden dendritic cells are shifted to ileocecal lymph nodes (iclns), where microbes are concentrated and an adequate immune response is triggered. hence, iclns are at a crucial position in immune anatomy and control processes of the local immune system. pathological alterations in iclns, such as reactive hyperplasia, lymphadenitis purulenta, or granulomatosa, can harbor a multitude of pathogens and commensals, posing a potential zoonotic risk in animal production. the aim of this study wa ...201424141125
single-cell analyses revealed transfer ranges of incp-1, incp-7, and incp-9 plasmids in a soil bacterial community.the conjugative transfer ranges of three different plasmids of the incompatibility groups incp-1 (pbp136), incp-7 (pcar1), and incp-9 (nah7) were investigated in soil bacterial communities by culture-dependent and culture-independent methods. pseudomonas putida, a donor of each plasmid, was mated with soil bacteria, and green fluorescent protein (gfp), encoded on the plasmid, was used as a reporter protein for successful transfer. gfp-expressing transconjugants were detected and separated at the ...201424141122
production of medium chain length fatty alcohols from glucose in escherichia coli.metabolic engineering offers the opportunity to produce a wide range of commodity chemicals that are currently derived from petroleum or other non-renewable resources. microbial synthesis of fatty alcohols is an attractive process because it can control the distribution of chain lengths and utilize low cost fermentation substrates. specifically, primary alcohols with chain lengths of 12 to 14 carbons have many uses in the production of detergents, surfactants, and personal care products. the cur ...201324141053
toxicity of perfluorooctanoic acid to pseudomonas putida in the aquatic environment.perfluorinated compounds are potential persistent organic pollutants which attracted much concerns in recent years. thus relevant toxicity data of perfluorooctanoic acid (pfoa) are vitally important for identification of possible risk in the aquatic environment. in the present study, the acute toxic effect of pfoa in the absence and presence of either chromium (iii) or tetra butyl ammonium (tba) towards pseudomonas putida in the aquatic environment was investigated by microcalorimetry. the therm ...201324140521
a bacterial swimmer with two alternating speeds of propagation.we recorded large data sets of swimming trajectories of the soil bacterium pseudomonas putida. like other prokaryotic swimmers, p. putida exhibits a motion pattern dominated by persistent runs that are interrupted by turning events. an in-depth analysis of their swimming trajectories revealed that the majority of the turning events is characterized by an angle of ϕ1 = 180° (reversals). to a lesser extent, turning angles of ϕ2 = 0° are also found. remarkably, we observed that, upon a reversal, th ...201324138867
removal of 3-methylindole by lactic acid bacteria in vitro.3-methylindole (3mi) is a substance with an unpleasant odor that is found in intact male pigs and is known to negatively affect consumers of pork. the growth of four strains of lactic acid bacteria [lactobacillus brevis 1.12 (l. brevis 1.12), l. plantarum 102, l. casei 6103 and l. plantarum atcc8014] in incubation medium with 3mi was studied. the four strains were tested for their ability to remove 3mi from the medium. the growth of l. brevis 1.12 remained steady as the levels of 3mi increased 3 ...201324137302
a new bioseed for determination of wastewater biodegradability: analysis of the experimental procedure.a new bioassay proposed in the patent p201300029 was applied to a pre-treated wastewater containing a mixture of commercial pesticides to simulate a recalcitrant industrial wastewater in order to determine its biodegradability. the test uses a mixture of standardized inoculum of the lyophilized bacteria pseudomonas putida with the proper proportion of salts and minerals. the results highlight that biodegradation efficiency can be calculated using a gross parameter (chemical oxygen demand (cod)) ...201424136578
genetic engineering of pseudomonas putida kt2440 for rapid and high-yield production of vanillin from ferulic acid.vanillin is one of the most important flavoring agents used today. that is why many efforts have been made on biotechnological production from natural abundant substrates. in this work, the nonpathogenic pseudomonas putida strain kt2440 was genetically optimized to convert ferulic acid to vanillin. deletion of the vanillin dehydrogenase gene (vdh) was not sufficient to prevent vanillin degradation. additional inactivation of a molybdate transporter, identified by transposon mutagenesis, led to a ...201424136472
involvement of antioxidant defense system in solvent tolerance of pseudomonas putida bcnu 106.the highly solvent-tolerant bacterium pseudomonas sp. bcnu 106 was investigated to elucidate the solvent tolerance under specific culture conditions with the presence of solvents and its adaptive mechanisms to those conditions with reference to the antioxidant system. when exposed to 10% toluene, pseudomonas sp. bcnu 106 increased the generation of reactive oxygen species assessed by monitoring the oxidation of 2',7'-dichlorofluorescein. typical antioxidant enzymes (viz. catalase, superoxide dis ...201424136354
gene expression profiling of pseudomonas putida f1 after exposure to aromatic hydrocarbon in soil by using proteome analysis.pseudomonas putida f1 can metabolize toluene, ethylbenzene, and benzene for growth. previously, we identified proteins involved in the utilization of these compounds by p. putida f1 through culture in liquid media. however, it was unclear whether laboratory analysis of bacterial activity and catabolism accurately reflected the soil environment. we identified proteins involved in the degradation of toluene, ethylbenzene, and benzene growth in soil using two-dimensional gel electrophoresis (2-de) ...201324136143
selective steroid oxyfunctionalisation by cyp154c5, a bacterial cytochrome p450.cytochrome p450 monooxygenases--able to regio- and stereoselectively hydroxylate non-activated carbon atoms--are important enzymes for the synthesis of valuable intermediates in the production of steroid hormones in the pharmaceutical industry. however, up to now only a few bacterial enzymes able to hydroxylate steroids have been reported. cyp154c5 from nocardia farcinica ifm 10152, a bacterial p450 monooxygenase, was previously shown to convert testosterone to 16α-hydroxytestosterone. since the ...201324134652
proteinaceous determinants of surface colonization in bacteria: bacterial adhesion and biofilm formation from a protein secretion perspective.bacterial colonization of biotic or abiotic surfaces results from two quite distinct physiological processes, namely bacterial adhesion and biofilm formation. broadly speaking, a biofilm is defined as the sessile development of microbial cells. biofilm formation arises following bacterial adhesion but not all single bacterial cells adhering reversibly or irreversibly engage inexorably into a sessile mode of growth. among molecular determinants promoting bacterial colonization, surface proteins a ...201324133488
recent advances in optical biosensors for environmental monitoring and early warning.the growing number of pollutants requires the development of innovative analytical devices that are precise, sensitive, specific, rapid, and easy-to-use to meet the increasing demand for legislative actions on environmental pollution control and early warning. optical biosensors, as a powerful alternative to conventional analytical techniques, enable the highly sensitive, real-time, and high-frequency monitoring of pollutants without extensive sample preparation. this article reviews important a ...201324132229
co-culture with listeria monocytogenes within a dual-species biofilm community strongly increases resistance of pseudomonas putida to benzalkonium chloride.biofilm formation is a phenomenon occurring almost wherever microorganisms and surfaces exist in close proximity. this study aimed to evaluate the possible influence of bacterial interactions on the ability of listeria monocytogenes and pseudomonas putida to develop a dual-species biofilm community on stainless steel (ss), as well as on the subsequent resistance of their sessile cells to benzalkonium chloride (bc) used in inadequate (sub-lethal) concentration (50 ppm). the possible progressive a ...201324130873
genome analysis coupled with physiological studies reveals a diverse nitrogen metabolism in methylocystis sp. strain sc2.methylocystis sp. strain sc2 can adapt to a wide range of methane concentrations. this is due to the presence of two isozymes of particulate methane monooxygenase exhibiting different methane oxidation kinetics. to gain insight into the underlying genetic information, its genome was sequenced and found to comprise a 3.77 mb chromosome and two large plasmids.201324130670
plant purine nucleoside catabolism employs a guanosine deaminase required for the generation of xanthosine in arabidopsis.purine nucleotide catabolism is common to most organisms and involves a guanine deaminase to convert guanine to xanthine in animals, invertebrates, and microorganisms. using metabolomic analysis of mutants, we demonstrate that arabidopsis thaliana uses an alternative catabolic route employing a highly specific guanosine deaminase (gsda) not reported from any organism so far. the enzyme is ubiquitously expressed and deaminates exclusively guanosine and 2'-deoxyguanosine but no other aminated puri ...201324130159
the crystal structure of pseudomonas putida azoreductase - the active site revisited.the enzymatic degradation of azo dyes begins with the reduction of the azo bond. in this article, we report the crystal structures of the native azoreductase from pseudomonas putida met94 (ppazor) (1.60 å), of ppazor in complex with anthraquinone-2-sulfonate (1.50 å), and of ppazor in complex with reactive black 5 dye (1.90 å). these structures reveal the residues and subtle changes that accompany substrate binding and release. such changes highlight the fine control of access to the catalytic s ...201324127652
expression, purification, and characterization of formaldehyde dehydrogenase from pseudomonas aeruginosa.as a member of zinc-containing medium-chain alcohol dehydrogenase family, formaldehyde dehydrogenase (fdh) can oxidize toxic formaldehyde to less active formate with nad(+) as a cofactor and exists in both prokaryotes and eukaryotes. most fdhs are well known to be glutathione-dependent in the catalysis of formaldehyde oxidation, but the enzyme from pseudomonas putida is an exception, which is independent of glutathione. to identify novel glutathione-independent fdhs from other bacterial strains ...201324125754
a tale of two machines: a review of the blast meeting, tucson, az, 20-24 january 2013.since its inception, bacterial locomotion and signal transduction (blast) meetings have been the place to exchange and share the latest developments in the field of bacterial signal transduction and motility. at the 12th blast meeting, held last january in tucson, az, researchers from all over the world met to report and discuss progress in diverse aspects of the field. the majority of these advances, however, came at the level of atomic level structures and their associated mechanisms. this was ...201324125587
a tale of two machines: a review of the blast meeting, tucson, az, 20-24 january 2013.since its inception, bacterial locomotion and signal transduction (blast) meetings have been the place to exchange and share the latest developments in the field of bacterial signal transduction and motility. at the 12th blast meeting, held last january in tucson, az, researchers from all over the world met to report and discuss progress in diverse aspects of the field. the majority of these advances, however, came at the level of atomic level structures and their associated mechanisms. this was ...201324125587
genomic and functional analyses of the 2-aminophenol catabolic pathway and partial conversion of its substrate into picolinic acid in burkholderia xenovorans lb400.2-aminophenol (2-ap) is a toxic nitrogen-containing aromatic pollutant. burkholderia xenovorans lb400 possess an amn gene cluster that encodes the 2-ap catabolic pathway. in this report, the functionality of the 2-aminophenol pathway of b. xenovorans strain lb400 was analyzed. the amnrjbacdfehg cluster located at chromosome 1 encodes the enzymes for the degradation of 2-aminophenol. the absence of haba and habb genes in lb400 genome correlates with its no growth on nitrobenzene. rt-pcr analyses ...201324124510
the novel bacterial n-demethylase pdmab is responsible for the initial step of n,n-dimethyl-substituted phenylurea herbicide degradation.the environmental fate of phenylurea herbicides has received considerable attention in recent decades. the microbial metabolism of n,n-dimethyl-substituted phenylurea herbicides can generally be initiated by mono-n-demethylation. in this study, the molecular basis for this process was revealed. the pdmab genes in sphingobium sp. strain ybl2 were shown to be responsible for the initial mono-n-demethylation of commonly used n,n-dimethyl-substituted phenylurea herbicides. pdmab is the oxygenase com ...201324123738
model based evaluation of a contaminant plume development under aerobic and anaerobic conditions in 2d bench-scale tank experiments.the influence of transverse mixing on competitive aerobic and anaerobic biodegradation of a hydrocarbon plume was investigated using a two-dimensional, bench-scale flow-through laboratory tank experiment. in the first part of the experiment aerobic degradation of increasing toluene concentrations was carried out by the aerobic strain pseudomonas putida f1. successively, ethylbenzene (injected as a mixture of unlabeled and fully deuterium-labeled isotopologues) substituted toluene; nitrate was ad ...201424122285
the crc protein inhibits the production of polyhydroxyalkanoates in pseudomonas putida under balanced carbon/nitrogen growth conditions.pseudomonas putida synthesizes polyhydroxyalkanoates (phas) as storage compounds. pha synthesis is more active when the carbon source is in excess and the nitrogen source is limiting, but can also occur at a lower rate under balanced carbon/nitrogen ratios. this work shows that pha synthesis is controlled by the crc global regulator, a protein that optimizes carbon metabolism by inhibiting the expression of genes involved in the use of non-preferred carbon sources. crc acts post-transcriptionall ...201424118893
aerobic degradation of n-methyl-4-nitroaniline (mna) by pseudomonas sp. strain fk357 isolated from soil.n-methyl-4-nitroaniline (mna) is used as an additive to lower the melting temperature of energetic materials in the synthesis of insensitive explosives. although the biotransformation of mna under anaerobic condition has been reported, its aerobic microbial degradation has not been documented yet. a soil microcosms study showed the efficient aerobic degradation of mna by the inhabitant soil microorganisms. an aerobic bacterium, pseudomonas sp. strain fk357, able to utilize mna as the sole carbon ...201324116023
the effect of polyphosphate kinase gene deletion on polyhydroxyalkanoate accumulation and carbon metabolism in pseudomonas putida kt2440.the primary enzyme involved in polyphosphate (polyp) synthesis, polyp kinase (ppk), has been deleted in pseudomonas putida kt2440. this has resulted in a threefold to sixfold reduction in polyhydroxyalkanoate (pha) accumulation compared with the wild type under conditions of nitrogen limitation, with either temperature or oxidative (h2o2) stress, when grown on glucose. the accumulation of pha by δppk mutant was the same as the wild type under nitrogen-limiting growth conditions. there was no dif ...201324115625
new methods for the isolation and characterization of biofilm-persistent mutants in pseudomonas putida.here we describe two new methods for the genetic characterization of bacterial biofilm development. first, we have designed a microtitre dish-based approach for high-throughput screening of pseudomonas putida mutants showing increased biofilm under dispersal conditions. using this method, nine such biofilm-persistent mutants, bearing transposon insertions in four loci: lapg, bifa, mvab and dksa, were isolated. second, we have developed a serial dilution-based scheme to monitor biofilm developmen ...201324115618
expression and purification of recombinant human serum albumin from selectively terminable transgenic rice.human serum albumin (hsa) is widely utilized for medical purposes and biochemical research. transgenic rice has proved to be an attractive bioreactor for mass production of recombinant hsa (rhsa). however, transgene spread is a major environmental and food safety concern for transgenic rice expressing proteins of medical value. this study aimed to develop a selectively terminable transgenic rice line expressing hsa in rice seeds, and a simple process for recovery and purification of rhsa for eco ...201324101203
resource availability and competition shape the evolution of survival and growth ability in a bacterial community.resource availability is one of the main factors determining the ecological dynamics of populations or species. fluctuations in resource availability can increase or decrease the intensity of resource competition. resource availability and competition can also cause evolutionary changes in life-history traits. we studied how community structure and resource fluctuations affect the evolution of fitness related traits using a two-species bacterial model system. replicated populations of serratia m ...201324098791
effects of indole-3-acetic acid on the transcriptional activities and stress tolerance of bradyrhizobium japonicum.a genome-wide transcriptional profile of bradyrhizobium japonicum, the nitrogen-fixing endosymbiont of the soybean plant, revealed differential expression of approximately 15% of the genome after a 1 mm treatment with the phytohormone indole-3-acetic acid (iaa). a total of 1,323 genes were differentially expressed (619 up-regulated and 704 down-regulated) at a two-fold cut off with q value ≤ 0.05. general stress response genes were induced, such as those involved in response to heat, cold, oxida ...201324098533
pseudomonas putida alka and alkb proteins comprise different defense systems for the repair of alkylation damage to dna - in vivo, in vitro, and in silico studies.alkylating agents introduce cytotoxic and/or mutagenic lesions to dna bases leading to induction of adaptive (ada) response, a mechanism protecting cells against deleterious effects of environmental chemicals. in escherichia coli, the ada response involves expression of four genes: ada, alka, alkb, and aidb. in pseudomonas putida, the organization of ada regulon is different, raising questions regarding regulation of ada gene expression. the aim of the presented studies was to analyze the role o ...201324098441
description of a riboflavin biosynthetic gene variant prevalent in the phylum proteobacteria.riboflavin (vitamin b2) is the precursor of flavin mononucleotide and flavin adenine dinucleotide, which are cofactors essential for a host of intracellular redox reactions. microorganisms synthesize flavins de novo to fulfill nutritional requirements, but it is becoming increasingly clear that flavins play a wider role in cellular physiology than was previously appreciated. flavins mediate diverse processes beyond the cytoplasmic membrane, including iron acquisition, extracellular respiration, ...201324097946
diverse broad-host-range plasmids from freshwater carry few accessory genes.broad-host-range self-transferable plasmids are known to facilitate bacterial adaptation by spreading genes between phylogenetically distinct hosts. these plasmids typically have a conserved backbone region and a variable accessory region that encodes host-beneficial traits. we do not know, however, how well plasmids that do not encode accessory functions can survive in nature. the goal of this study was to characterize the backbone and accessory gene content of plasmids that were captured from ...201324096417
complete nucleotide sequence of the self-transmissible tol plasmid pd2rt provides new insight into arrangement of toluene catabolic plasmids.in the present study we report the complete nucleotide sequence of the toluene catabolic plasmid pd2rt of pseudomonas migulae strain d2rt isolated from baltic sea water. the pd2rt is 129,894 base pairs in size with an average g+c content of 53.75%. a total of 135 open reading frames (orfs) were predicted to encode proteins, among them genes for catabolism of toluene, plasmid replication, maintenance and conjugative transfer. orfs encoding proteins with putative functions in stress response, tran ...201324095800
phacos, a functionalized bacterial polyester with bactericidal activity against methicillin-resistant staphylococcus aureus.biomaterial-associated infections represent a significant clinical problem, and treatment of these microbial infections is becoming troublesome due to the increasing number of antibiotic-resistant strains. here, we report a naturally functionalized bacterial polyhydroxyalkanoate (phacos) with antibacterial properties. we demonstrate that phacos selectively and efficiently inhibits the growth of methicillin-resistant staphylococcus aureus (mrsa) both in vitro and in vivo. this ability has been as ...201324094939
phacos, a functionalized bacterial polyester with bactericidal activity against methicillin-resistant staphylococcus aureus.biomaterial-associated infections represent a significant clinical problem, and treatment of these microbial infections is becoming troublesome due to the increasing number of antibiotic-resistant strains. here, we report a naturally functionalized bacterial polyhydroxyalkanoate (phacos) with antibacterial properties. we demonstrate that phacos selectively and efficiently inhibits the growth of methicillin-resistant staphylococcus aureus (mrsa) both in vitro and in vivo. this ability has been as ...201324094939
photocatalytic bacterial inactivation by tio2-coated surfaces.the aim of this study was the evaluation of the photoactivated antibacterial activity of titanium dioxide (tio2)-coated surfaces. bacterial inactivation was evaluated using tio2-coated petri dishes. the experimental conditions optimized with petri dishes were used to test the antibacterial effect of tio2-coated ceramic tiles. the best antibacterial effect with petri dishes was observed at 180, 60, 30 and 20 min of exposure for escherichia coli, staphylococcus aureus, pseudomonas putida and liste ...201324090112
proposal of a quantitative pcr-based protocol for an optimal pseudomonas aeruginosa detection in patients with cystic fibrosis.the lung of patients with cystic fibrosis (cf) is particularly sensitive to pseudomonas aeruginosa. this bacterium plays an important role in the poor outcome of cf patients. during the disease progress, first acquisition of p. aeruginosa is the key-step in the management of cf patients. quantitative pcr (qpcr) offers an opportunity to detect earlier the first acquisition of p. aeruginosa by cf patients. given the lack of a validated protocol, our goal was to find an optimal molecular protocol f ...201324088260
host adaptation is contingent upon the infection route taken by pathogens.evolution of pathogen virulence is affected by the route of infection. also, alternate infection routes trigger different physiological responses on hosts, impinging on host adaptation and on its interaction with pathogens. yet, how route of infection may shape adaptation to pathogens has not received much attention at the experimental level. we addressed this question through the experimental evolution of an outbred drosophila melanogaster population infected by two different routes (oral and s ...201324086131
a bacterial quercetin oxidoreductase quoa-mediated perturbation in the phenylpropanoid metabolic network increases lignification with a concomitant decrease in phenolamides in arabidopsis.metabolic perturbations by a gain-of-function approach provide a means to alter steady states of metabolites and query network properties, while keeping enzyme complexes intact. a combination of genetic and targeted metabolomics approach was used to understand the network properties of phenylpropanoid secondary metabolism pathways. a novel quercetin oxidoreductase, quoa, from pseudomonas putida, which converts quercetin to naringenin, thus effectively reversing the biosynthesis of quercetin thro ...201324085580
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