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molecular analysis of the subgingival microbiota in health and disease.this investigation provides molecular analyses of the periodontal microbiota in health and disease. subgingival samples from 47 volunteers with healthy gingivae or clinically diagnosed chronic periodontitis were characterized by pcr-denaturing gradient gel electrophoresis (dgge) with primers specific for the v2-v3 region of the eubacterial 16s rrna gene. a hierarchical dendrogram was constructed from band patterns. all unique pcr amplicons (dgge bands) were sequenced for identity. samples were a ...200717085691
neutrophil apoptosis: a marker of disease severity in sepsis and sepsis-induced acute respiratory distress syndrome.apoptosis of neutrophils (polymorphonuclear neutrophils [pmns]) may limit inflammatory injury in sepsis and acute respiratory distress syndrome (ards), but the relationship between the severity of sepsis and extent of pmn apoptosis and the effect of superimposed ards is unknown. the objective of this study was to correlate neutrophil apoptosis with the severity of sepsis and sepsis-induced ards.200617092345
novel macrolide resistance module carried by the incp-1beta resistance plasmid prsb111, isolated from a wastewater treatment plant.the macrolide resistance plasmid prsb111 was isolated from bacteria residing in the final effluents of a wastewater treatment plant. the 47-kb plasmid confers resistance to azithromycin, clarithromycin, erythromycin, roxithromycin, and tylosin when it is carried by pseudomonas sp. strain b13 and is very similar to prototype incp-1beta plasmid pb3, which was previously isolated from an activated-sludge bacterial community of a wastewater treatment plant. the two plasmids differ in their accessory ...200717101677
a gene cluster involved in degradation of substituted salicylates via ortho cleavage in pseudomonas sp. strain mt1 encodes enzymes specifically adapted for transformation of 4-methylcatechol and 3-methylmuconate.pseudomonas sp. strain mt1 has recently been reported to degrade 4- and 5-chlorosalicylate by a pathway assumed to consist of a patchwork of reactions comprising enzymes of the 3-oxoadipate pathway. genes encoding the initial steps in the degradation of salicylate and substituted derivatives were now localized and sequenced. one of the gene clusters characterized (sal) showed a novel gene arrangement, with sala, encoding a salicylate 1-hydroxylase, being clustered with salcd genes, encoding muco ...200717172348
x-ray structure of a hydroxylase-regulatory protein complex from a hydrocarbon-oxidizing multicomponent monooxygenase, pseudomonas sp. ox1 phenol hydroxylase.phenol hydroxylase (ph) belongs to a family of bacterial multicomponent monooxygenases (bmms) with carboxylate-bridged diiron active sites. included are toluene/o-xylene (tomo) and soluble methane (smmo) monooxygenase. ph hydroxylates aromatic compounds, but unlike smmo, it cannot oxidize alkanes despite having a similar dinuclear iron active site. important for activity is formation of a complex between the hydroxylase and a regulatory protein component. to address how structural features of bm ...200617176061
mechanistic studies of the flavoenzyme tryptophan 2-monooxygenase: deuterium and 15n kinetic isotope effects on alanine oxidation by an l-amino acid oxidase.tryptophan 2-monooxygenase (tmo) from pseudomonas savastanoi catalyzes the oxidative decarboxylation of l-tryptophan during the biosynthesis of indoleacetic acid. structurally and mechanistically, the enzyme is a member of the family of l-amino acid oxidases. deuterium and 15n kinetic isotope effects were used to probe the chemical mechanism of l-alanine oxidation by tmo. the primary deuterium kinetic isotope effect was ph independent over the ph range 6.5-10, with an average value of 6.0 +/- 0. ...200617176107
identification of potential cepr regulated genes using a cep box motif-based search of the burkholderia cenocepacia genome.the burkholderia cenocepacia cepir quorum sensing system has been shown to positively and negatively regulate genes involved in siderophore production, protease expression, motility, biofilm formation and virulence. in this study, two approaches were used to identify genes regulated by the cepir quorum sensing system. transposon mutagenesis was used to create lacz promoter fusions in a cepi mutant that were screened for differential expression in the presence of n-acylhomoserine lactones. a bioi ...200617187664
adaptation of pseudomonas sp. strain 7-6 to quaternary ammonium compounds and their degradation via dual pathways.pseudomonas sp. strain 7-6, isolated from active sludge obtained from a wastewater facility, utilized a quaternary ammonium surfactant, n-dodecyltrimethylammonium chloride (dtac), as its sole carbon, nitrogen, and energy source. when initially grown in the presence of 10 mm dtac medium, the isolate was unable to degrade dtac. the strain was cultivated in gradually increasing concentrations of the surfactant until continuous exposure led to high tolerance and biodegradation of the compound. based ...200717261523
the role of laterally transferred genes in adaptive evolution.bacterial genomes develop new mechanisms to tide them over the imposing conditions they encounter during the course of their evolution. acquisition of new genes by lateral gene transfer may be one of the dominant ways of adaptation in bacterial genome evolution. lateral gene transfer provides the bacterial genome with a new set of genes that help it to explore and adapt to new ecological niches.200717288581
proteoglycan synthesis by human corneal explants submitted to laser in situ keratomileusis (lasik).to evaluate the acute effects of laser in situ keratomileusis (lasik) upon the synthesis of proteoglycans (pgs) and collagen fibril organization in human corneal explants.200717293778
generation by a widely applicable approach of a hybrid dioxygenase showing improved oxidation of polychlorobiphenyls.recently, a sequence-based approach has been developed for the fast isolation and characterization of class ii aryl-hydroxylating dioxygenase activities (s. kahl and b. hofer, microbiology 149:1475-1481, 2003). it comprises the pcr amplification of segments of alpha subunit genes of unknown sequence that encode the catalytic center and their fusion with sequences of the bpha gene cluster of burkholderia xenovorans lb400. one of the resulting chimeric enzymes, harboring the core segment of a diox ...200717322323
structural investigations of the ferredoxin and terminal oxygenase components of the biphenyl 2,3-dioxygenase from sphingobium yanoikuyae b1.the initial step involved in oxidative hydroxylation of monoaromatic and polyaromatic compounds by the microorganism sphingobium yanoikuyae strain b1 (b1), previously known as sphingomonas yanoikuyae strain b1 and beijerinckia sp. strain b1, is performed by a set of multiple terminal rieske non-heme iron oxygenases. these enzymes share a single electron donor system consisting of a reductase and a ferredoxin (bpdo-fb1). one of the terminal rieske oxygenases, biphenyl 2,3-dioxygenase (bpdo-ob1), ...200717349044
outcome of life-threatening malaria in african children requiring endotracheal intubation.little is known about children undergoing critical care for malaria. the purpose of this survey was to evaluate the outcome in african children requiring endotracheal intubation for life-threatening malaria.200717470294
substrate specificity of a long-chain alkylamine-degrading pseudomonas sp isolated from activated sludge.a bacterium strain bert, which utilizes primary long-chain alkylamines as nitrogen, carbon and energy source, was isolated from activated sludge. this rod-shaped motile, gram-negative strain was identified as a pseudomonas sp. the substrate spectrum of this pseudomonas strain bert includes primary alkylamines with alkyl chains ranging from c(3) to c(18), and dodecyl-1,3-diaminopropane. amines with alkyl chains ranging from 8 to 14 carbons were the preferred substrates. growth on dodecanal, dodec ...200817492358
biochemical and crystallographic analysis of substrate binding and conformational changes in acetyl-coa synthetase.the adenylate-forming enzymes, including acyl-coa synthetases, the adenylation domains of non-ribosomal peptide synthetases (nrps), and firefly luciferase, perform two half-reactions in a ping-pong mechanism. we have proposed a domain alternation mechanism for these enzymes whereby, upon completion of the initial adenylation reaction, the c-terminal domain of these enzymes undergoes a 140 degrees rotation to perform the second thioester-forming half-reaction. structural and kinetic data of mutan ...200717497934
metabolomic and genetic analysis of biomarkers for peroxisome proliferator-activated receptor alpha expression and activation.peroxisome proliferator-activated receptor alpha (pparalpha) is a nuclear receptor with manifold effects on intermediary metabolism. to define a set of urinary biomarkers that could be used to determine the efficacy of pparalpha agonists, a metabolomic investigation was undertaken in wild-type and pparalpha-null mice fed for 2 wk either a regular diet or a diet containing the pparalpha ligand wy-14,643 ([4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio] acetic acid), and their urine was analyzed by u ...200717550978
finding intermediates in the o2 activation pathways of non-heme iron oxygenases.intermediates in the reaction cycle of an oxygenase are usually very informative with respect to the chemical mechanism of o 2 activation and insertion. however, detection of these intermediates is often complicated by their short lifetime and the regulatory mechanism of the enzyme designed to ensure specificity. here, the methods used to detect the intermediates in an extradiol dioxygenase, a rieske cis-dihydrodiol dioxygenase, and soluble methane monooxygenase are discussed. the methods includ ...200717567087
a survey of orphan enzyme activities.using computational database searches, we have demonstrated previously that no gene sequences could be found for at least 36% of enzyme activities that have been assigned an enzyme commission number. here we present a follow-up literature-based survey involving a statistically significant sample of such "orphan" activities. the survey was intended to determine whether sequences for these enzyme activities are truly unknown, or whether these sequences are absent from the public sequence databases ...200717623104
dissimilatory iodate reduction by marine pseudomonas sp. strain sct.bacterial iodate (io(3)(-)) reduction is poorly understood largely due to the limited number of available isolates as well as the paucity of information about key enzymes involved in the reaction. in this study, an iodate-reducing bacterium, designated strain sct, was newly isolated from marine sediment slurry. sct is phylogenetically closely related to the denitrifying bacterium pseudomonas stutzeri and reduced 200 microm iodate to iodide (i(-)) within 12 h in an anaerobic culture containing 10 ...200717644635
hydroxyatrazine n-ethylaminohydrolase (atzb): an amidohydrolase superfamily enzyme catalyzing deamination and dechlorination.hydroxyatrazine [2-(n-ethylamino)-4-hydroxy-6-(n-isopropylamino)-1,3,5-triazine] n-ethylaminohydrolase (atzb) is the sole enzyme known to catalyze the hydrolytic conversion of hydroxyatrazine to n-isopropylammelide. atzb, therefore, serves as the point of intersection of multiple s-triazine biodegradative pathways and is completely essential for microbial growth on s-triazine herbicides. here, atzb was cloned from pseudomonas sp. strain adp and its product was purified to homogeneity and charact ...200717660279
a novel sampling method for the investigation of gut mirobiota.in order to characterize the qualitative and quantitative microorganisms in different sites of the lower digestive tract (ldt) in healthy volunteers, a specific technique was developed for collecting mucous of the distal ileum, colon and rectum.200717663516
biochemical and genetic investigation of initial reactions in aerobic degradation of the bile acid cholate in pseudomonas sp. strain chol1.bile acids are surface-active steroid compounds with toxic effects for bacteria. recently, the isolation and characterization of a bacterium, pseudomonas sp. strain chol1, growing with bile acids as the carbon and energy source was reported. in this study, initial reactions of the aerobic degradation pathway for the bile acid cholate were investigated on the biochemical and genetic level in strain chol1. these reactions comprised a-ring oxidation, activation with coenzyme a (coa), and beta-oxida ...200717693490
similar enzymes, different structures: phthalate dioxygenase is an alpha3alpha3 stacked hexamer, not an alpha3beta3 trimer like "normal" rieske oxygenases.phthalate dioxygenase (pdo) is a member of a class of bacterial oxygenases that contain both rieske [2fe-2s] and fe(ii) mononuclear centers. recent crystal structures of several rieske dioxygenases showed that they exist as alpha(3)beta(3) multimers with subunits arranged head-to-tail in alpha and beta stacked planar rings. the structure of pdo, which consists of only alpha-subunits, remains to be solved. although similar to other rieske dioxygenases in many aspects, pdo was shown to differ in t ...200717764654
c-myc interacts with trf1/pin2 and regulates telomere length.telomere, the end of linear chromosome, is protected by dna-protein complexes. these complexes cap the linear chromosome and play an important role in the maintenance of genomic stability. trf1/pin2, a double-stranded dna-binding protein is known to regulate telomere length by not only protecting telomere but also blocking the access of telomerase to telomere in cis. to better understand the mechanism through which trf1/pin2 regulates telomere length, we performed the yeast two-hybrid screening ...200717765874
conformationally constrained analogues of 2-arachidonoylglycerol.novel monocyclic analogues of 2-arachidonoylglycerol (2-ag) were designed in order to explore the pharmacophoric conformations of this endocannabinoid ligand at the key cannabinergic proteins. all 2-arachidonoyl esters of 1,2,3-cyclohexanetriol [meso-7 (am5504), (+/-)-8 (am5503), and meso-9 (am5505)] were synthesized by regioselective acylation of 2,3-dihydroxycyclohexanone followed by selective reductions. the optically active isomers (+)-8 (am4434) and (-)-8 (am4435) were synthesized from (2s, ...200717826996
plp catabolism: identification of the 4-pyridoxic acid dehydrogenase gene in mesorhizobium loti maff303099.the function of the mlr6793 gene from mesorhizobium loti maff303099 has been identified. this gene encodes 4-pyridoxic acid dehydrogenase, an enzyme involved in the catabolism of plp (vitamin b6). this enzyme was overexpressed in escherichia coli and characterized. 4-pyridoxic acid dehydrogenase is a 33kda protein that catalyzes the four electron oxidation of 4-pyridoxic acid to 3-hydroxy-2-methylpyridine-4,5-dicarboxylate, using nicotinamide adenine dinucleotide as a cofactor. the k cat for nad ...200717868768
production of l-carnitine by secondary metabolism of bacteria.abstract: the increasing commercial demand for l-carnitine has led to a multiplication of efforts to improve its production with bacteria. the use of different cell environments, such as growing, resting, permeabilized, dried, osmotically stressed, freely suspended and immobilized cells, to maintain enzymes sufficiently active for l-carnitine production is discussed in the text. the different cell states of enterobacteria, such as escherichia coli and proteus sp., which can be used to produce l- ...200717910757
a comprehensive set of plasmids for vanillate- and xylose-inducible gene expression in caulobacter crescentus.caulobacter crescentus is widely used as a powerful model system for the study of prokaryotic cell biology and development. analysis of this organism is complicated by a limited selection of tools for genetic manipulation and inducible gene expression. this study reports the identification and functional characterization of a vanillate-regulated promoter (p(van)) which meets all requirements for application as a multi-purpose expression system in caulobacter, thus complementing the established x ...200717959646
gene identification and structural characterization of the pyridoxal 5'-phosphate degradative protein 3-hydroxy-2-methylpyridine-4,5-dicarboxylate decarboxylase from mesorhizobium loti maff303099.the function of the mlr6791 gene from mesorhizobium loti maff303099 has been identified. this gene encodes 3-hydroxy-2-methylpyridine-4,5-dicarboxylate decarboxylase (hmpddc), an enzyme involved in the catabolism of pyridoxal 5'-phosphate (vitamin b6). this enzyme was overexpressed in escherichia coli and characterized. hmpddc is a 26 kda protein that catalyzes the decarboxylation of 3-hydroxy-2-methylpyridine-4,5-dicarboxylate to 3-hydroxy-2-methylpyridine-5-carboxylate. the km and kcat were fo ...200717973403
a novel caffeine dehydrogenase in pseudomonas sp. strain cbb1 oxidizes caffeine to trimethyluric acid.a unique heterotrimeric caffeine dehydrogenase was purified from pseudomonas sp. strain cbb1. this enzyme oxidized caffeine to trimethyluric acid stoichiometrically and hydrolytically, without producing hydrogen peroxide. the enzyme was not nad(p)(+) dependent; coenzyme q(0) was the preferred electron acceptor. the enzyme was specific for caffeine and theobromine and showed no activity with xanthine.200817981969
nadh availability limits asymmetric biocatalytic epoxidation in a growing recombinant escherichia coli strain.styrene can efficiently be oxidized to (s)-styrene oxide by recombinant escherichia coli expressing the styrene monooxygenase genes styab from pseudomonas sp. strain vlb120. targeting microbial physiology during whole-cell redox biocatalysis, we investigated the interdependency of styrene epoxidation, growth, and carbon metabolism on the basis of mass balances obtained from continuous two-liquid-phase cultures. full induction of styab expression led to growth inhibition, which could be attenuate ...200818192422
sub-grouping and sub-functionalization of the rifin multi-copy protein family.parasitic protozoans possess many multicopy gene families which have central roles in parasite survival and virulence. the number and variability of members of these gene families often make it difficult to predict possible functions of the encoded proteins. the families of extra-cellular proteins that are exposed to a host immune response have been driven via immune selection to become antigenically variant, and thereby avoid immune recognition while maintaining protein function to establish a ...200818197962
efficient display of active lipase lipb52 with a pichia pastoris cell surface display system and comparison with the lipb52 displayed on saccharomyces cerevisiae cell surface.for industrial bioconversion processes, the utilization of surface-displayed lipase in the form of whole-cell biocatalysts is more advantageous, because the enzymes are displayed on the cell surface spontaneously, regarded as immobilized enzymes.200818221563
functional role of the additional domains in inulosucrase (isla) from leuconostoc citreum cw28.inulosucrase (isla) from leuconostoc citreum cw28 belongs to a new subfamily of multidomain fructosyltransferases (ftfs), containing additional domains from glucosyltransferases. it is not known what the function of the additional domains in this subfamily is.200818237396
versatility of biological non-heme fe(ii) centers in oxygen activation reactions.oxidase and oxygenase enzymes allow the use of relatively unreactive o2 in biochemical reactions. many of the mechanistic strategies used in nature for this key reaction are represented within the 2-histidine-1-carboxylate facial triad family of non-heme fe(ii)-containing enzymes. the open face of the metal coordination sphere opposite the three endogenous ligands participates directly in the reaction chemistry. here, data from several studies are presented showing that reductive o2 activation w ...200818277980
"hot cores" in proteins: comparative analysis of the apolar contact area in structures from hyper/thermophilic and mesophilic organisms.a wide variety of stabilizing factors have been invoked so far to elucidate the structural basis of protein thermostability. these include, amongst the others, a higher number of ion-pairs interactions and hydrogen bonds, together with a better packing of hydrophobic residues. it has been frequently observed that packing of hydrophobic side chains is improved in hyperthermophilic proteins, when compared to their mesophilic counterparts. in this work, protein crystal structures from hyper/thermop ...200818312638
survey of bacterial diversity in chronic wounds using pyrosequencing, dgge, and full ribosome shotgun sequencing.chronic wound pathogenic biofilms are host-pathogen environments that colonize and exist as a cohabitation of many bacterial species. these bacterial populations cooperate to promote their own survival and the chronic nature of the infection. few studies have performed extensive surveys of the bacterial populations that occur within different types of chronic wound biofilms. the use of 3 separate16s-based molecular amplifications followed by pyrosequencing, shotgun sanger sequencing, and denatur ...200818325110
novel aromatic ring-hydroxylating dioxygenase genes from coastal marine sediments of patagonia.polycyclic aromatic hydrocarbons (pahs), widespread pollutants in the marine environment, can produce adverse effects in marine organisms and can be transferred to humans through seafood. our knowledge of pah-degrading bacterial populations in the marine environment is still very limited, and mainly originates from studies of cultured bacteria. in this work, genes coding catabolic enzymes from pah-biodegradation pathways were characterized in coastal sediments of patagonia with different levels ...200818366740
a new classification system for bacterial rieske non-heme iron aromatic ring-hydroxylating oxygenases.rieske non-heme iron aromatic ring-hydroxylating oxygenases (rhos) are multi-component enzyme systems that are remarkably diverse in bacteria isolated from diverse habitats. since the first classification in 1990, there has been a need to devise a new classification scheme for these enzymes because many rhos have been discovered, which do not belong to any group in the previous classification. here, we present a scheme for classification of rhos reflecting new sequence information and interactio ...200818387195
biphenyl dioxygenase from an arctic isolate is not cold adapted.biphenyl dioxygenase from the psychrotolerant bacterium pseudomonas sp. strain cam-1 (bpdo(cam-1)) was purified and found to have an apparent k(cat) for biphenyl of 1.1 +/- 0.1 s(-1) (mean +/- standard deviation) at 4 degrees c. in contrast, bpdo(lb400) from the mesophile burkholderia xenovorans lb400 had no detectable activity at this temperature. at 57 degrees c, the half-life of the bpdo(cam-1) oxygenase was less than half that of the bpdo(lb400) oxygenase. nevertheless, bpdo(cam-1) appears t ...200818424535
action for child survival: elimination of haemophilus influenzae type b meningitis in uganda.to guide immunization policy, we determined the public health benefit of introducing haemophilus influenzae type b (hib) vaccine in uganda and estimated the vaccine effectiveness.200818438518
the majority of in vitro macrophage activation exhibited by extracts of some immune enhancing botanicals is due to bacterial lipoproteins and lipopolysaccharides.we have identified potent monocyte/macrophage activating bacterial lipoproteins within commonly used immune enhancing botanicals such as echinacea, american ginseng and alfalfa sprouts. these bacterial lipoproteins, along with lipopolysaccharides, were substantially more potent than other bacterially derived components when tested in in vitro monocyte/macrophage activation systems. in experiments using raw 264.7 and mouse peritoneal macrophages the majority (85-98%) of the activity within extrac ...200818486914
genes and enzymes of azetidine-2-carboxylate metabolism: detoxification and assimilation of an antibiotic.l-(-)-azetidine-2-carboxylate (ac) is a toxic, natural product analog of l-proline. this study revealed the genes and biochemical strategy employed by pseudomonas sp. strain a2c to detoxify and assimilate ac as its sole nitrogen source. the gene region from pseudomonas sp. strain a2c required for detoxification was cloned into escherichia coli and sequenced. the 7.0-kb region contained eight identifiable genes. four encoded putative transporters or permeases for gamma-amino acids or drugs. anoth ...200818487339
the binary protein interactome of treponema pallidum--the syphilis spirochete.protein interaction networks shed light on the global organization of proteomes but can also place individual proteins into a functional context. if we know the function of bacterial proteins we will be able to understand how these species have adapted to diverse environments including many extreme habitats. here we present the protein interaction network for the syphilis spirochete treponema pallidum which encodes 1,039 proteins, 726 (or 70%) of which interact via 3,649 interactions as revealed ...200818509523
x-ray structure of ill2, an auxin-conjugate amidohydrolase from arabidopsis thaliana.the plant hormone indole-3-acetic acid (iaa) is the most abundant natural auxin involved in many aspects of plant development and growth. the iaa levels in plants are modulated by a specific group of amidohydrolases from the peptidase m20d family that release the active hormone from its conjugated storage forms. here, we describe the x-ray crystal structure of iaa-amino acid hydrolase iaa-leucine resistantlike gene 2 (ill2) from arabidopsis thaliana at 2.0 a resolution. ill2 preferentially hydro ...200918543330
crystal structure of a carbonyl reductase from candida parapsilosis with anti-prelog stereospecificity.a novel short-chain (s)-1-phenyl-1,2-ethanediol dehydrogenase (scr) from candida parapsilosis exhibits coenzyme specificity for nadph over nadh. it catalyzes an anti-prelog type reaction to reduce 2-hydroxyacetophenone into (s)-1-phenyl-1,2-ethanediol. the coding gene was overexpressed in escherichia coli and the purified protein was crystallized. the crystal structure of the apo-form was solved to 2.7 a resolution. this protein forms a homo-tetramer with a broken 2-2-2 symmetry. the overall fol ...200818566346
a novel transposon, tn6009, composed of a tn916 element linked with a staphylococcus aureus mer operon.the aim of this study was to characterize a novel conjugative transposon tn6009 composed of a tn916 linked to a staphylococcus aureus mer operon in representative gram-positive and gram-negative bacteria isolated in nigeria and portugal.200818583328
the plant-associated bacterium stenotrophomonas rhizophila expresses a new enzyme for the synthesis of the compatible solute glucosylglycerol.the rhizobacterium stenotrophomonas rhizophila accumulates the compatible solutes glucosylglycerol (gg) and trehalose under salt stress conditions. the complete gene for the gg synthesis enzyme was cloned and sequenced. this enzyme from s. rhizophila represented a novel fusion protein composed of a putative c-terminal gg-phosphate synthase domain and an n-terminal putative gg-phosphate phosphatase domain, which was named ggpps. a similar gene was cloned from pseudomonas sp. strain oa146. the ggp ...200818586931
structural characterization of a 140 degrees domain movement in the two-step reaction catalyzed by 4-chlorobenzoate:coa ligase.members of the adenylate-forming family of enzymes play a role in the metabolism of halogenated aromatics and of short, medium, and long chain fatty acids, as well as in the biosynthesis of menaquinone, peptide antibiotics, and peptide siderophores. this family includes a subfamily of acyl- and aryl-coa ligases that catalyze thioester synthesis through two half-reactions. a carboxylate substrate first reacts with atp to form an acyl-adenylate. subsequent to the release of the product pp i, the e ...200818620418
poly-3-hydroxybutyrate synthase from the periplasm of escherichia coli.this is the first report of a poly-3-hydroxybutyrate (phb) synthase in escherichia coli. the enzyme was isolated from the periplasm using ammonium sulfate fractionation, hydrophobic, and size-exclusion chromatography and identified by lc/ms/ms as ydcs, a component of a putative abc transporter. green fluorescent protein-tagged ydcs, purified by 2d native gel electrophoresis, also exhibited phb synthase activity. optimal conditions for enzyme activity were 37 degrees c, ph 6.8-7.5, 100 mm kcl. k( ...200818640095
identification and characterization of bacterial cutinase.cutinase, which exists in both fungi and bacteria, catalyzes the cleavage of the ester bonds of cutin. fungal cutinases have been extensively studied, however, reports on bacterial cutinases have been limited due to the lack of knowledge concerning the identity of their open reading frames. in the present study, the cutinase from thermobifida fusca was induced by cutin and purified to homogeneity by following p-nitrophenyl butyrate hydrolyzing activity. peptide mass fingerprinting analysis of th ...200818658138
the dape-encoded n-succinyl-l,l-diaminopimelic acid desuccinylase from haemophilus influenzae contains two active-site histidine residues.the catalytic and structural properties of the h67a and h349a dape-encoded n-succinyl-l,l-diaminopimelic acid desuccinylase (dape) from haemophilus influenzae were investigated. on the basis of sequence alignment with the carboxypeptidase from pseudomonas sp. strain rs-16, both h67 and h349 were predicted to be zn(ii) ligands. the h67a dape enzyme exhibited a decreased catalytic efficiency (180-fold) compared with wild-type (wt) dape towards n-succinyldiaminopimelic acid. no catalytic activity w ...200918712420
a genome-scale metabolic reconstruction of pseudomonas putida kt2440: ijn746 as a cell factory.pseudomonas putida is the best studied pollutant degradative bacteria and is harnessed by industrial biotechnology to synthesize fine chemicals. since the publication of p. putida kt2440's genome, some in silico analyses of its metabolic and biotechnology capacities have been published. however, global understanding of the capabilities of p. putida kt2440 requires the construction of a metabolic model that enables the integration of classical experimental data along with genomic and high-through ...200818793442
unconventional serine proteases: variations on the catalytic ser/his/asp triad configuration.serine proteases comprise nearly one-third of all known proteases identified to date and play crucial roles in a wide variety of cellular as well as extracellular functions, including the process of blood clotting, protein digestion, cell signaling, inflammation, and protein processing. their hallmark is that they contain the so-called "classical" catalytic ser/his/asp triad. although the classical serine proteases are the most widespread in nature, there exist a variety of "nonclassical" serine ...200818824507
intermediate in the o-o bond cleavage reaction of an extradiol dioxygenase.the reactive oxy intermediate of the catalytic cycle of extradiol aromatic ring-cleaving dioxygenases is formed by binding the catecholic substrate and o2 in adjacent ligand positions of the active site metal [usually fe(ii)]. this intermediate and the following fe(ii)-alkylperoxo intermediate resulting from oxygen attack on the substrate have been previously characterized in a crystal of homoprotocatechuate 2,3-dioxygenase (hpcd). here a subsequent intermediate in which the o-o bond is broken t ...200818826259
structure of a sigma28-regulated nonflagellar virulence protein from campylobacter jejuni.campylobacter jejuni, a gram-negative motile bacterium, is a leading cause of human gastrointestinal infections. although the mechanism of c.jejuni-mediated enteritis appears to be multifactorial, flagella play complex roles in the virulence of this human pathogen. cj0977 is a recently identified virulence factor in c. jejuni and is expressed by a sigma(28) promoter that controls late genes in the flagellar regulon. a cj0977 mutant strain is fully motile but significantly reduced in the invasion ...200818835274
fast quantitative determination of microbial rhamnolipids from cultivation broths by atr-ftir spectroscopy.vibrational spectroscopic techniques are becoming increasingly important and popular because they have the potential to provide rapid and convenient solutions to routine analytical problems. using these techniques, a variety of substances can be characterized, identified and also quantified rapidly.200818840269
variability in in vitro macrophage activation by commercially diverse bulk echinacea plant material is predominantly due to bacterial lipoproteins and lipopolysaccharides.we previously reported that the majority of in vitro monocyte/macrophage activation exhibited by extracts of echinacea and other botanicals depends upon bacterial lipopolysaccharides and braun-type bacterial lipoproteins. we determined the contribution made by these bacterial components to the overall immune-enhancing activity detected in e. purpurea and e. angustifolia bulk root and aerial material obtained from six major growers/suppliers in north america. substantial variation in activity (up ...200818942831
pruning nature: biodiversity-derived discovery of novel sodium channel blocking conotoxins from conus bullatus.described herein is a general approach to identify novel compounds using the biodiversity of a megadiverse group of animals; specifically, the phylogenetic lineage of the venomous gastropods that belong to the genus conus ("cone snails"). cone snail biodiversity was exploited to identify three new mu-conotoxins, buiiia, buiiib and buiiic, encoded by the fish-hunting species conus bullatus. buiiia, buiiib and buiiic are strikingly divergent in their amino acid composition compared to previous mu- ...200818950653
pruning nature: biodiversity-derived discovery of novel sodium channel blocking conotoxins from conus bullatus.described herein is a general approach to identify novel compounds using the biodiversity of a megadiverse group of animals; specifically, the phylogenetic lineage of the venomous gastropods that belong to the genus conus ("cone snails"). cone snail biodiversity was exploited to identify three new mu-conotoxins, buiiia, buiiib and buiiic, encoded by the fish-hunting species conus bullatus. buiiia, buiiib and buiiic are strikingly divergent in their amino acid composition compared to previous mu- ...200818950653
the transcriptional repressor protein nsrr senses nitric oxide directly via a [2fe-2s] cluster.the regulatory protein nsrr, a member of the rrf2 family of transcription repressors, is specifically dedicated to sensing nitric oxide (no) in a variety of pathogenic and non-pathogenic bacteria. it has been proposed that no directly modulates nsrr activity by interacting with a predicted [fe-s] cluster in the nsrr protein, but no experimental evidence has been published to support this hypothesis. here we report the purification of nsrr from the obligate aerobe streptomyces coelicolor. we demo ...200818989365
resolving genetic functions within microbial populations: in situ analyses using rrna and mrna stable isotope probing coupled with single-cell raman-fluorescence in situ hybridization.prokaryotes represent one-half of the living biomass on earth, with the vast majority remaining elusive to culture and study within the laboratory. as a result, we lack a basic understanding of the functions that many species perform in the natural world. to address this issue, we developed complementary population and single-cell stable isotope ((13)c)-linked analyses to determine microbial identity and function in situ. we demonstrated that the use of rrna/mrna stable isotope probing (sip) rec ...200918997025
mechanism of extradiol aromatic ring-cleaving dioxygenases.the extradiol aromatic ring-cleaving dioxygenases activate molecular oxygen by binding both o(2) and the catecholic substrate to a reduced active site metal, generally fe(ii). progress has been made in understanding the mechanism of this reaction through the combined use of kinetic, computational, biomimetic, structural, and diagnostic chemical approaches. it appears that o(2) is activated by accepting an electron transferred from the substrate through the metal, thereby simultaneously activatin ...200819007887
the 1.6 a crystal structure of mycobacterium smegmatis mshc: the penultimate enzyme in the mycothiol biosynthetic pathway.mycobacterium smegmatis mshc catalyzes the atp-dependent condensation of glcn-ins and l-cysteine to form l-cys-glcn-ins, the penultimate step in mycothiol biosynthesis. attempts to crystallize the native, full-length mshc have been unsuccessful. however, incubation of the enzyme with the cysteinyl adenylate analogue, 5'-o-[n-(l-cysteinyl)-sulfamonyl]adenosine (csa), followed by a 24-h limited trypsin proteolysis yielded an enzyme preparation that readily crystallized. the three-dimensional struc ...200819053270
molecular determinants of the regioselectivity of toluene/o-xylene monooxygenase from pseudomonas sp. strain ox1.bacterial multicomponent monooxygenases (bmms) are a heterogeneous family of di-iron monooxygenases which share the very interesting ability to hydroxylate aliphatic and/or aromatic hydrocarbons. each bmm possesses defined substrate specificity and regioselectivity which match the metabolic requirements of the strain from which it has been isolated. pseudomonas sp. strain ox1, a strain able to metabolize o-, m-, and p-cresols, produces the bmm toluene/o-xylene monooxygenase (tomo), which convert ...200919074607
anenpi: identification and annotation of analogous enzymes.enzymes are responsible for the catalysis of the biochemical reactions in metabolic pathways. analogous enzymes are able to catalyze the same reactions, but they present no significant sequence similarity at the primary level, and possibly different tertiary structures as well. they are thought to have arisen as the result of independent evolutionary events. a detailed study of analogous enzymes may reveal new catalytic mechanisms, add information about the origin and evolution of biochemical pa ...200819091081
a modeling study by response surface methodology and artificial neural network on culture parameters optimization for thermostable lipase production from a newly isolated thermophilic geobacillus sp. strain arm.thermostable bacterial lipases occupy a place of prominence among biocatalysts owing to their novel, multifold applications and resistance to high temperature and other operational conditions. the capability of lipases to catalyze a variety of novel reactions in both aqueous and nonaqueous media presents a fascinating field for research, creating interest to isolate novel lipase producers and optimize lipase production. the most important stages in a biological process are modeling and optimizat ...200819105837
lumbar periradicular abscess mimicking a fragmented lumbar disc herniation : an unusual case.we herein describe the case of a focal spontaneous spinal epidural abscess who was initially diagnosed to have a free fragment of a lumbar disc. a 71-year-old woman presented with history of low back and right leg pain. magnetic resonance imaging suggested a peripherally enhancing free fragment extending down from s1 nerve root axilla. preoperative laboratory investigation showed elevation of c-reactive protein (crp), erythrocyte sedimentation rate (esr) levels. she was taken for surgery and a f ...200819137084
the mechanisms of human hotdog-fold thioesterase 2 (hthem2) substrate recognition and catalysis illuminated by a structure and function based analysis.the focus of this paper is the hotdog-fold thioesterase them2 from human (hthem2; swiss-prot entry q9npj3 ). in an earlier communication (cheng, z., song, f., shan, x., wei, z., wang, y., dunaway-mariano, d., and gong, w. (2006) crystal structure of human thioesterase superfamily member 2, biochem. biophys. res. commun. 349, 172-177) we reported the apo crystal structure of hthem2. herein, we report the results of an extensive hthem2 substrate screen, the structure determination of hthem2 comple ...200919170545
export of recombinant proteins in escherichia coli using abc transporter with an attached lipase abc transporter recognition domain (lard).abstract:200919178697
structural analysis reveals dna binding properties of rv2827c, a hypothetical protein from mycobacterium tuberculosis.tuberculosis (tb) is a major global health threat caused by mycobacterium tuberculosis (mtb). it is further fueled by the hiv pandemic and by increasing incidences of multidrug resistant mtb-strains. rv2827c, a hypothetical protein from mtb, has been implicated in the survival of mtb in the macrophages of the host. the three-dimensional structure of rv2827c has been determined by the three-wavelength anomalous diffraction technique using bromide-derivatized crystals and refined to a resolution o ...200919184528
identification and characterization of catabolic para-nitrophenol 4-monooxygenase and para-benzoquinone reductase from pseudomonas sp. strain wbc-3.pseudomonas sp. strain wbc-3 utilizes para-nitrophenol (pnp) as a sole source of carbon, nitrogen, and energy. in order to identify the genes involved in this utilization, we cloned and sequenced a 12.7-kb fragment containing a conserved region of nad(p)h:quinone oxidoreductase genes. of the products of the 13 open reading frames deduced from this fragment, pnpa shares 24% identity to the large component of a 3-hydroxyphenylacetate hydroxylase from pseudomonas putida u and pnpb is 58% identical ...200919218392
investigation of atomic level patterns in protein--small ligand interactions.shape complementarity and non-covalent interactions are believed to drive protein-ligand interaction. to date protein-protein, protein-dna, and protein-rna interactions were systematically investigated, which is in contrast to interactions with small ligands. we investigate the role of covalent and non-covalent bonds in protein-small ligand interactions using a comprehensive dataset of 2,320 complexes.200919221587
a rapid crosstalk of human gammadelta t cells and monocytes drives the acute inflammation in bacterial infections.vgamma9/vdelta2 t cells are a minor subset of t cells in human blood and differ from other t cells by their immediate responsiveness to microbes. we previously demonstrated that the primary target for vgamma9/vdelta2 t cells is (e)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (hmb-pp), an essential metabolite produced by a large range of pathogens. here we wished to study the consequence of this unique responsiveness in microbial infection. the majority of peripheral vgamma9/vdelta2 t cells share ...200919229322
the sensor kinase tods operates by a multiple step phosphorelay mechanism involving two autokinase domains.expression of the pseudomonas putida tod operon, which encodes enzymes for toluene metabolism, takes place from the p(todx) promoter and is mediated by the tods/todt two component system. the sensor kinase tods has a complex domain arrangement containing two functional modules, each harboring a sensor- and an autokinase domain and separated by a receiver domain. based on site-directed mutagenesis of phosphoaccepting his-190, asp-500, and his-760 and in vitro transphosphorylation experiments with ...200919240030
a tn5051-like mer-containing transposon identified in a heavy metal tolerant strain achromobacter sp. ao22.achromobacter sp. ao22 (formerly alcaligenes sp. ao22), a bacterial strain isolated from a lead-contaminated industrial site in australia, was previously found to be resistant to moderate to high levels of mercury, copper and other heavy metals. however, the nature and location of the genetic basis for mercuric ion resistance in this strain, had not been previously identified.200919284535
synthesis of rhamnolipid biosurfactant and mode of hexadecane uptake by pseudomonas species.abstract:200919284586
the pha depolymerase engineering database: a systematic analysis tool for the diverse family of polyhydroxyalkanoate (pha) depolymerases.polyhydroxyalkanoates (phas) can be degraded by many microorganisms using intra- or extracellular pha depolymerases. pha depolymerases are very diverse in sequence and substrate specificity, but share a common alpha/beta-hydrolase fold and a catalytic triad, which is also found in other alpha/beta-hydrolases.200919296857
studies of a ring-cleaving dioxygenase illuminate the role of cholesterol metabolism in the pathogenesis of mycobacterium tuberculosis.mycobacterium tuberculosis, the etiological agent of tb, possesses a cholesterol catabolic pathway implicated in pathogenesis. this pathway includes an iron-dependent extradiol dioxygenase, hsac, that cleaves catechols. immuno-compromised mice infected with a deltahsac mutant of m. tuberculosis h37rv survived 50% longer than mice infected with the wild-type strain. in guinea pigs, the mutant disseminated more slowly to the spleen, persisted less successfully in the lung, and caused little pathol ...200919300498
structure of the plp degradative enzyme 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase from mesorhizobium loti maff303099 and its mechanistic implications.a vitamin b(6) degradative pathway has recently been identified and characterized in mesorhizobium loti maff303099. one of the enzymes on this pathway, 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase (mhpco), is a flavin-dependent enzyme and catalyzes the oxidative ring-opening of 2-methyl-3-hydroxypyridine-5-carboxylic acid to form e-2-(acetamino-methylene)succinate. the gene for this enzyme has been cloned, and the corresponding protein has been overexpressed in escherichia coli and pur ...200919317437
increased diversity of predacious bdellovibrio-like organisms (blos) as a function of eutrophication in kumaon lakes of india.predation by bdellovibrio-like organisms (blos) results in bacterial community succession in aquatic ecosystems. the effects of nutrient loading on the distribution and phylogeny of blos remain largely unknown. to this end, we present our findings on blo diversity from four north-indian lakes that are variable in their trophic status; nainital is eutrophic, both, bhimtal and naukuchiatal are mesotrophic and sattal remains oligotrophic, respectively. initially, total heterotrophic bacteria and bl ...200919319600
the mechanism of domain alternation in the acyl-adenylate forming ligase superfamily member 4-chlorobenzoate: coenzyme a ligase.4-chlorobenzoate:coa ligase (cbl) belongs to the adenylate-forming family of enzymes that catalyze a two-step reaction to first activate a carboxylate substrate as an adenylate and then transfer the carboxylate to the pantetheine group of either coenzyme a or an acyl-carrier protein. the active site is located at the interface of a large n-terminal domain and a smaller c-terminal domain. crystallographic structures have been determined at multiple steps along the reaction pathway and form the ba ...200919320426
constitutive and regulated expression vectors to construct polyphosphate deficient bacteria.inorganic polyphosphate (polyp), a polymer of tens or hundreds of phosphate residues linked by atp-like bonds, is found in all organisms and performs a wide variety of functions. polyp is synthesized in bacterial cells by the actions of polyphosphate kinases (ppk1 and ppk2) and degraded by an exopolyphosphatase (ppx). bacterial cells with polyp deficiencies are impaired in many structural and important cellular functions such as motility, quorum sensing, biofilm formation and virulence. knockout ...200919323822
performance of a yeast-mediated biological fuel cell.saccharomyces cerevisiae present in common baker's yeast was used in a microbial fuel cell in which glucose was the carbon source. methylene blue was used as the electronophore in the anode compartment, while potassium ferricyanide and methylene blue were tested as electron acceptors in the cathode compartment. microbes in a mediator-free environment were used as the control. the experiment was performed in both open and closed circuit configurations under different loads ranging from 100 komega ...200819325724
co2 production as an indicator of biofilm metabolism.biofilms are important in aquatic nutrient cycling and microbial proliferation. in these structures, nutrients like carbon are channeled into the production of extracellular polymeric substances or cell division; both are vital for microbial survival and propagation. the aim of this study was to assess carbon channeling into cellular or noncellular fractions in biofilms. growing in tubular reactors, biofilms of our model strain pseudomonas sp. strain ct07 produced cells to the planktonic phase f ...200919346353
medical bioremediation of age-related diseases.abstract: catabolic insufficiency in humans leads to the gradual accumulation of a number of pathogenic compounds associated with age-related diseases, including atherosclerosis, alzheimer's disease, and macular degeneration. removal of these compounds is a widely researched therapeutic option, but the use of antibodies and endogenous human enzymes has failed to produce effective treatments, and may pose risks to cellular homeostasis. another alternative is "medical bioremediation," the use of m ...200919358742
massive comparative genomic analysis reveals convergent evolution of specialized bacteria.genome size and gene content in bacteria are associated with their lifestyles. obligate intracellular bacteria (i.e., mutualists and parasites) have small genomes that derived from larger free-living bacterial ancestors; however, the different steps of bacterial specialization from free-living to intracellular lifestyle have not been studied comprehensively. the growing number of available sequenced genomes makes it possible to perform a statistical comparative analysis of 317 genomes from bacte ...200919361336
16s rrna gene-based analysis of fecal microbiota from preterm infants with and without necrotizing enterocolitis.neonatal necrotizing enterocolitis (nec) is an inflammatory intestinal disorder affecting preterm infants. intestinal bacteria have an important function; however no causative pathogen has been identified. the purpose of this study was to determine if there are differences in microbial patterns that may be critical to the development of this disease. fecal samples from 20 preterm infants, 10 with nec and 10 matched controls (including 4 twin pairs) were obtained from patients in a single site le ...200919369970
two small ncrnas jointly govern virulence and transmission in legionella pneumophila.to transit from intra- to extracellular environments, legionella pneumophila differentiates from a replicative/non-virulent to a transmissive/virulent form using the two-component system leta/lets and the global repressor protein csra. while investigating how both regulators act co-ordinately we characterized two ncrnas, rsmy and rsmz, that link the leta/lets and csra regulatory networks. we demonstrate that leta directly regulates their expression and show that rsmy and rsmz are functional in e ...200919400772
biodegradation of the high explosive hexanitrohexaazaiso-wurtzitane (cl-20).the aerobic biodegradability of the high explosive cl-20 by activated sludge and the white rot fungus phanerochaete chrysosporium has been investigated. although activated sludge is not effective in degrading cl-20 directly, it can mineralize the alkaline hydrolysis products. phanerochaete chrysosporium degrades cl-20 in the presence of supplementary carbon and nitrogen sources. biodegradation studies were conducted using various nutrient media under diverse conditions. variables included the cl ...200919440524
redox-linked conformational dynamics in apoptosis-inducing factor.apoptosis-inducing factor (aif) is a bifunctional mitochondrial flavoprotein critical for energy metabolism and induction of caspase-independent apoptosis, whose exact role in normal mitochondria remains unknown. upon reduction with nadh, aif undergoes dimerization and forms tight, long-lived fadh(2)-nad charge-transfer complexes (ctc) that are proposed to be functionally important. to obtain a deeper insight into structure/function relations and redox mechanism of this vitally important protein ...200919447115
the stat pathway mediates late-phase immunity against plasmodium in the mosquito anopheles gambiae.the stat family of transcription factors activates expression of immune system genes in vertebrates. the ancestral stat gene (agstat-a) appears to have duplicated in the mosquito anopheles gambiae, giving rise to a second intronless stat gene (agstat-b), which we show regulates agstat-a expression in adult females. agstat-a participates in the transcriptional activation of nitric oxide synthase (nos) in response to bacterial and plasmodial infection. activation of this pathway, however, is not e ...200919454353
distal end of 105-125 loop--a putative reductase binding domain of phthalate dioxygenase.the phthalate dioxygenase system consists of the dioxygenase, pdo, which contains a rieske [2fe-2s] center and a fe(ii)-mononuclear center, and the reductase, pdr. involvement of the distal end of the 105-125 loop of pdo in its interaction with pdr was tested by substituting charged residues in the loop with alanines and by replacing the conserved tryptophan-94. compared to wild-type pdo, all variants had lower catalytic activity and the rieske centers were reduced more slowly by reduced pdr. th ...200919464996
analysis of proteins with the 'hot dog' fold: prediction of function and identification of catalytic residues of hypothetical proteins.the hot dog fold has been found in more than sixty proteins since the first report of its existence about a decade ago. the fold appears to have a strong association with fatty acid biosynthesis, its regulation and metabolism, as the proteins with this fold are predominantly coenzyme a-binding enzymes with a variety of substrates located at their active sites.200919473548
modeling of solvent-dependent conformational transitions in burkholderia cepacia lipase.the characteristic of most lipases is the interfacial activation at a lipid interface or in non-polar solvents. interfacial activation is linked to a large conformational change of a lid, from a closed to an open conformation which makes the active site accessible for substrates. while for many lipases crystal structures of the closed and open conformation have been determined, the pathway of the conformational transition and possible bottlenecks are unknown. therefore, molecular dynamics simula ...200919476626
identification of a novel self-sufficient styrene monooxygenase from rhodococcus opacus 1cp.sequence analysis of a 9-kb genomic fragment of the actinobacterium rhodococcus opacus 1cp led to identification of an open reading frame encoding a novel fusion protein, stya2b, with a putative function in styrene metabolism via styrene oxide and phenylacetic acid. gene cluster analysis indicated that the highly related fusion proteins of nocardia farcinica ifm10152 and arthrobacter aurescens tc1 are involved in a similar physiological process. whereas 413 amino acids of the n terminus of stya2 ...200919482928
caenorhabditis elegans genomic response to soil bacteria predicts environment-specific genetic effects on life history traits.with the post-genomic era came a dramatic increase in high-throughput technologies, of which transcriptional profiling by microarrays was one of the most popular. one application of this technology is to identify genes that are differentially expressed in response to different environmental conditions. these experiments are constructed under the assumption that the differentially expressed genes are functionally important in the environment where they are induced. however, whether differential e ...200919503598
inhibition of human folylpolyglutamate synthetase by diastereomeric phosphinic acid mimics of the tetrahedral intermediate.phosphorus-containing pseudopeptides, racemic at the c-terminal alpha-carbon, are potent mechanism-based inhibitors of folylpolyglutamate synthetase (fpgs). they are mimics of the tetrahedral intermediate postulated to form during fpgs-catalyzed biosynthesis of poly(gamma-l-glutamates). in the present paper, the fpgs inhibitory activity of each diastereomer coupled to three heterocycles is reported. the high r(f) pseudopeptide containing the 5,10-dideazatetrahydropteroyl (ddah(4)pte) heterocycle ...200919563773
arsenic accumulation by pseudomonas stutzeri and its response to some thiol chelators.the aim of this study is to examine arsenic accumulation by pseudomonas stutzeri and its response to some thiol chelators, dmps and miadmsa.200819568912
metagenomics reveals diversity and abundance of meta-cleavage pathways in microbial communities from soil highly contaminated with jet fuel under air-sparging bioremediation.the extradiol dioxygenase diversity of a site highly contaminated with aliphatic and aromatic hydrocarbons under air-sparging treatment was assessed by functional screening of a fosmid library in escherichia coli with catechol as substrate. the 235 positive clones from inserts of dna extracted from contaminated soil were equivalent to one extradiol dioxygenase-encoding gene per 3.6 mb of dna screened, indicating a strong selection for genes encoding this function. three subfamilies were identifi ...200919575758
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