Home About us Contact
|
Home About us Contact | ||
|
|
||
P. Putida (p + putida)
Selected AbstractsGas Diffusion Electrodes for Use in an Amperometric Enzyme BiosensorELECTROANALYSIS, Issue 21 2008Martin Hämmerle Abstract The preparation of gas diffusion electrodes and their use in an amperometric enzyme biosensor for the direct detection of a gaseous analyte is described. The gas diffusion electrodes are prepared by covering a PTFE membrane (thickness 250,,m, pore size 2,,m, porosity 35%) with gold, platinum, or a graphite/PTFE mixture. Gold and platinum are deposited by e-beam sputtering, whereas the graphite/PTFE layer is prepared by vacuum filtration of a respective aqueous suspension. These gas diffusion electrodes are exemplarily implemented as working electrodes in an amperometric biosensor for gaseous formaldehyde containing NAD-dependent formaldehyde dehydrogenase from P. putida [EC. 1.2.1.46] as enzyme and 1,2-naphthoquinone-4-sulfonic acid as electrochemical mediator. The resulting sensors are compared with regard to background current, signal noise, linear range, sensitivity, and detection limit. In this respect, sensors with gold or graphite/PTFE covered membranes outclass ones with platinum for this particular analyte and sensor configuration. [source] DNA sequence-based analysis of the Pseudomonas speciesENVIRONMENTAL MICROBIOLOGY, Issue 6 2010Magdalena Mulet Summary Partial sequences of four core ,housekeeping' genes (16S rRNA, gyrB, rpoB and rpoD) of the type strains of 107 Pseudomonas species were analysed in order to obtain a comprehensive view regarding the phylogenetic relationships within the Pseudomonas genus. Gene trees allowed the discrimination of two lineages or intrageneric groups (IG), called IG P. aeruginosa and IG P. fluorescens. The first IG P. aeruginosa, was divided into three main groups, represented by the species P. aeruginosa, P. stutzeri and P. oleovorans. The second IG was divided into six groups, represented by the species P. fluorescens, P. syringae, P. lutea, P. putida, P. anguilliseptica and P. straminea. The P. fluorescens group was the most complex and included nine subgroups, represented by the species P. fluorescens, P. gessardi, P. fragi, P. mandelii, P. jesseni, P. koreensis, P. corrugata, P. chlororaphis and P. asplenii. Pseudomonas rhizospherae was affiliated with the P. fluorescens IG in the phylogenetic analysis but was independent of any group. Some species were located on phylogenetic branches that were distant from defined clusters, such as those represented by the P. oryzihabitans group and the type strains P. pachastrellae, P. pertucinogena and P. luteola. Additionally, 17 strains of P. aeruginosa, ,P. entomophila', P. fluorescens, P. putida, P. syringae and P. stutzeri, for which genome sequences have been determined, have been included to compare the results obtained in the analysis of four housekeeping genes with those obtained from whole genome analyses. [source] Lack of CbrB in Pseudomonas putida affects not only amino acids metabolism but also different stress responses and biofilm developmentENVIRONMENTAL MICROBIOLOGY, Issue 6 2010Cristina I. Amador Summary The CbrAB two-component system has been described in certain species of Pseudomonads as a global regulatory system required for the assimilation of several amino acids (e.g. histidine, proline or arginine) as carbon or carbon and nitrogen sources. In this work, we used global gene expression and phenotypic analyses to characterize the roles of the CbrAB system in Pseudomonas putida. Our results show that CbrB is involved in coordination with the nitrogen control system activator, NtrC, in the uptake and assimilation of several amino acids. In addition, CbrB affects other carbon utilization pathways and a number of apparently unrelated functions, such as chemotaxis, stress tolerance and biofilm development. Based on these new findings, we propose that CbrB is a high-ranked element in the regulatory hierarchy of P. putida that directly or indirectly controls a variety of metabolic and behavioural traits required for adaptation to changing environmental conditions. [source] Temperature and pyoverdine-mediated iron acquisition control surface motility of Pseudomonas putidaENVIRONMENTAL MICROBIOLOGY, Issue 7 2007Miguel A. Matilla Summary Pseudomonas putida KT2440 is unable to swarm at its common temperature of growth in the laboratory (30°C) but exhibits surface motility similar to swarming patterns in other Pseudomonas between 18°C and 28°C. These motile cells show differentiation, consisting on elongation and the presence of surface appendages. Analysis of a collection of mutants to define the molecular determinants of this type of surface movement in KT2440 shows that while type IV pili and lipopolysaccharide O-antigen are requisites flagella are not. Although surface motility of flagellar mutants was macroscopically undistinguishable from that of the wild type, microscopy analysis revealed that these mutants move using a distinct mechanism to that of the wild-type strain. Mutants either in the siderophore pyoverdine (ppsD) or in the FpvA siderophore receptor were also unable to spread on surfaces. Motility in the ppsD strain was totally restored with pyoverdine and partially with the wild-type ppsD allele. Phenotype of the fpvA strain was not complemented by this siderophore. We discuss that iron influences surface motility and that it can be an environmental cue for swarming-like movement in P. putida. This study constitutes the first report assigning an important role to pyoverdine iron acquisition in en masse bacterial surface movement. [source] The ttgGHI solvent efflux pump operon of Pseudomonas putida DOT-T1E is located on a large self-transmissible plasmidENVIRONMENTAL MICROBIOLOGY, Issue 6 2007José J. Rodríguez-Herva Summary Pseudomonas putida DOT-T1E is a solvent-tolerant strain able to grow in the presence of > 1% (v/v) toluene in the culture medium. A set of multidrug efflux pumps have been found to play a major role in the tolerance of this bacterium to organic solvents (Rojas et al., J Bacteriol 183: 3967,3973). In the course of studies of the mechanisms underlying solvent tolerance in DOT-T1E, we isolated a spontaneous solvent-sensitive mutant derivative which had lost the genes encoding the TtgGHI efflux pump, the most important extrusion element in quantitative terms. Genomic comparisons between the mutant and its parental strain by microarray analysis revealed that in addition to the ttgVW-ttgGHI gene cluster, another group of genes, highly similar to those found in the Tn4653A and ISPpu12 transposable elements of the TOL plasmid pWW0 from P. putida mt-2, were also absent from this strain. Further analysis demonstrated that strain DOT-T1E harboured a large plasmid (named pGRT1) that was lost from the solvent-sensitive mutant. Mapping analysis revealed that the ttgVW-ttgGHI genes and the Tn4653A -like transposon are borne by the pGRT1 plasmid. Plasmid pGRT1 is highly stable and its frequency of loss is below 10,8 per cell per generation under a variety of growth conditions, including nutritional and physical stresses. The pGRT1 plasmid is self-transmissible, and its acquisition by the toluene-sensitive P. putida KT2440 and Pseudomonas aeruginosa PAO1 increased the recipient's tolerance to toluene up to levels similar to those exhibited by P. putida DOT-T1E. We discuss the importance and potential benefits of this plasmid for the development of bacteria with enhanced solvent tolerance, and its potential impact for bioremediation and whole-cell biotransformations. [source] Mutualistic symbiosis between Bursaphelenchus xylophilus and bacteria of the genus PseudomonasFOREST PATHOLOGY, Issue 5 2005B. G. Zhao Summary Interactions between the pine wood nematode (PWN), Bursaphelenchus xylophilus, and bacteria of the genus Pseudomonas were examined by cultivating axenic PWN and bacterial strains using callus of Pinus thunbergii. Ten (Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas cepacia and Pseudomonas spp.) of the 29 bacterial strains tested, significantly increased the reproduction of PWN. The rest of the bacteria (19 strains of 10 species) inhibited the reproduction of PWN completely. The growth of 18 of the 29 bacterial strains tested, including the 10 strains promoting PWN reproduction, was significantly increased by the presence of PWN. It indicated a mutualistic symbiotic relationship between PWN and the 10 bacterial strains in the genus Pseudomonas. The bacterial mutualistic symbionts are organisms, which may have co-evolved with PWN rather than being accidentally associated. The finding provides further evidence for our hypothesis that pine wilt disease is complex, induced by both PWN and associated phytotoxin-producing bacteria. Résumé Les interactions entre le nématode des pins Bursaphelenchus xylophilus (PWN) et des bactéries du genre Pseudomonas ont étéétudiées en cultivant de manière axénique PWN et des souches bactériennes sur des cals de Pinusthunbergii. Dix souches bactériennes (Pseudomonas fluorescens, P. putida, P. cepacia et Pseudomonas spp.) sur les 29 testées ont significativement augmenté la reproduction de PNW. Le reste des bactéries (19 souches de 10 espèces) ont complètement inhibé la reproduction de PNW. La croissance de 18 souches bactériennes sur 29, incluant les 10 favorisant la reproduction de PNW, a été significativement augmentée en présence de PNW. Ceci indique une relation symbiotique mutualiste entre PNW et 10 souches bactériennes du genre Pseudomonas. Les symbiontes bactériens mutualistes pourraient être des organismes ayant coévolué avec PNW plutôt qu'associés de façon fortuite. Ces observations renforcent l'hypothèse selon laquelle le flétrissement des pins est une maladie complexe induite par PWN en association avec des bactéries productrices de phytotoxines. Zusammenfassung Die Interaktionen zwischen dem Kiefernsplintholznematoden (PWN, Bursaphelenchus xylophilus) und Bakterien der Gattung Pseudomonas wurden in axenischen Kulturen des Nematoden mit verschiedenen Bakterienstämmen und Kallus von Pinus thunbergii untersucht. Zehn Bakterienstämme (Pseudomonas fluoreszens, Pseudomonas putida, Pseudomonas cepacia und Pseudomonas spp.) von 29 getesteten Isolaten erhöhten die Reproduktion des PWN signifikant. Die übrigen Isolate (19 Stämme von 10 Arten) hemmten die Vermehrung des Nematoden vollständig. Das Wachstum von 18 der 29 getesteten Bakterienstämme (inkl. der 10 Stämme, welche die Vermehrung des Nematoden förderten), wurde durch die Präsenz des Nematoden signifikant erhöht. Dieser Befund deutet auf eine mutualistische Beziehung zwischen dem PWN und 10 Pseudomonas -Isolaten hin. Die mutualistischen bakteriellen Symbionten dürften sich wahrscheinlich in Coevolution mit dem Nematoden entwickelt haben. Dieser Befund unterstützt die Hypothese, dass die Kiefernwelke eine Komplexkrankheit darstellt, die sowohl durch B. xylophilus als auch die damit assoziierten phytotoxinbildenden Bakterien ausgelöst wird. [source] Effects of some bacteria (Pseudomonas spp. and Erwinia herbicola) on in vitro growth of Piptoporus betulinusFOREST PATHOLOGY, Issue 6 2000K. Przyby Summary Bacteria including Pseudomonas putida, Pseudomonas fluorescens biovar I, Pseudomonas fluorescens biovar V, Pseudomonas aureofaciens and Erwinia herbicola were isolated from discoloured zones in birch trunks. Antagonistic effects of these bacteria to growth of Piptoporus betulinus mycelium were tested in vitro, both in dual culture and using bacterial cell-free culture filtrates. In dual cultures, P. putida was most effective at inhibiting mycelial growth of Piptoporus betulinus. Filtrates of P. putida inhibited growth of P. betulinus mycelium irrespective of filtrate concentration, incubation time of bacteria and timing of recording mycelium growth. The strongest antagonistic effect (inhibition of fungal growth) was observed on a medium containing 80% of sterile filtrate obtained from 15-day-old bacterial cultures. The highest stimulating effect on mycelium growth was noted on medium containing 80% filtrate obtained from 7-day-old E. herbicola cultures. Résumé Des bactéries, Pseudomonas putida, Pseudomonas fluorescens biovar I, Pseudomonas fluorescens biovar V, Pseudomonas aureofaciens et Erwinia herbicola, ont été isolées de zones colorées de troncs de bouleau. Les effets antagonistes de ces bactéries sur la croissance mycélienne de Piptoporus betulinus ont étéévalués in vitro, en cultures doubles et à partir de filtrats bactériens. En cultures doubles, P. putida a été le plus inhibiteur de la croissance du P. betulinus. Les filtrats de P. putida inhibaient la croissance quel que soit la concentration du filtrat, la durée d'incubation de la bactérie, et le délai dans lequel la croissance mycélienne était mesurée. L'effet inhibiteur le plus fort a été observé sur un milieu contenant 80% de filtrat stérile obtenu de cultures bactériennes de 15 jours. L'effet stimulant le plus fort a été noté sur un milieu contenant 80% d'un filtrat obtenu de cultures de 7 jours de E. herbicola. Zusammenfassung Verschiedene Bakterienarten (Pseudomonas putida, Pseudomonas fluorescens Biovar I, Pseudomonas fluorescens Biovar V, Pseudomonas aureofaciens und E. herbicola) wurden aus verfärbtem Holz in Birkenstämmen isoliert. Antagonistische Effekte dieser Bakterien gegenüber Myzel von Piptoporus betulinus wurden in vitroüberprüft (Dualkulturen und bakterienzellfreie Kulturfiltrate). In Dualkulturen zeigte P. putida den stärksten Hemmeffekt auf das Myzelwachstum von P. betulinus. Filtrate von P. putida hemmten das Wachstum von P. betulinus, unabhängig von der Filtratkonzentration, der Inkubationszeit der Bakterien und dem Zeitpunkt der Messung des Myzelwachstums. Der antagonistische Effekt (Hemmung des Myzelwachstums) war am ausgeprägtesten auf einem Medium, das 80% Sterilfiltrat von 15 Tage alten Bakterienkulturen enthielt. Der stärkste Stimulationseffekt auf das Myzelwachstum wurde auf einem Medium beobachtet, welches 80% Filtrat von sieben Tage alten E. herbicola -Kulturen enthielt. [source] Biocontrol of Meloidogyne incognita on tomato using antagonistic fungi, plant-growth-promoting rhizobacteria and cattle manurePEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 9 2009Zaki A Siddiqui Abstract BACKGROUND: Biocontrol achieved by a single biocontrol agent is generally inconsistent under field conditions. The aim of the present study was to increase the competitiveness and efficacy of biocontrol agents by using them together with cattle manure. RESULTS: The effects of antagonistic fungi [Aspergillus niger v. Teigh., Paecilomyces lilacinus (Thom) Samson and Penicillium chrysogenum Thom] and plant-growth-promoting rhizobacteria (PGPR) [Azotobacter chroococcum Beijer., Bacillus subtilis (Ehrenberg) Cohn and Pseudomonas putida (Trev.) Mig.] were assessed with cattle manure on the growth of tomato and on the reproduction of Meloidogyne incognita (Kof. & White) Chitwood. Application of antagonistic fungi and PGPR alone and in combination with cattle manure resulted in a significant increase in the growth of nematode-inoculated plants. The highest increase (79%) in the growth of nematode-inoculated plants was observed when P. putida was used with cattle manure, followed by use of P. lilacinus plus cattle manure. Paecilomyces lilacinus resulted in a high reduction in galling and nematode multiplication, followed by P. putida, B. subtilis, A. niger, A. chroococcum and P. chrysogenum. The combined use of P. lilacinus with cattle manure resulted in a maximum reduction in galling and nematode multiplication. CONCLUSION: Application of P. lilacinus or P. putida with cattle manure was useful to achieve greater biocontrol of M. incognita on tomato. Copyright © 2009 Society of Chemical Industry [source] Structure of 2-keto-3-deoxy-6-phosphogluconate (KDPG) aldolase from Pseudomonas putidaACTA CRYSTALLOGRAPHICA SECTION D, Issue 8 2003B. J. Bell 2-Keto-3-deoxy-6-phosphogluconate (KDPG) aldolase from Pseudomonas putida is a key enzyme in the Entner,Doudoroff pathway which catalyses the cleavage of KDPG via a class I Schiff-base mechanism. The crystal structure of this enzyme has been refined to a crystallographic residual R = 17.1% (Rfree = 21.4%). The N-terminal helix caps one side of the torus of the (,,)8 -barrel and the active site is located on the opposite, carboxylic side of the barrel. The Schiff-base-forming Lys145 is coordinated by a sulfate (or phosphate) ion and two solvent water molecules. The interactions that stabilize the trimer are predominantly hydrophobic, with the exception of the cyclically permuted bonds formed between Glu132,OE1 of one molecule and Thr129,OG1 of a symmetry-equivalent molecule. Except for the N-terminal helix, the structure of KDPG aldolase from P. putida closely resembles the structure of the homologous enzyme from Escherichia coli. [source] Fluorene and phenanthrene uptake by Pseudomonas putida ATCC 17514: Kinetics and physiological aspectsBIOTECHNOLOGY & BIOENGINEERING, Issue 3 2005Ana C. Rodrigues Abstract Pseudomonas putida ATCC 17514 was used as a model strain to investigate the characteristics of bacterial growth in the presence of solid fluorene and phenanthrene. Despite the lower water-solubility of phenanthrene, P. putida degraded this polycyclic aromatic hydrocarbon (PAH) at a maximum observed rate of 1.4 ± 0.1 mg L,1 h,1, higher than the apparent degradation rate of fluorene, 0.8 ± 0.07 mg L,1 h,1. The role of physiological processes on the biodegradation of these PAHs was analyzed and two different uptake strategies were identified. Zeta potential measurements revealed that phenanthrene-grown cells were slightly more negatively charged (,57.5 ± 4.7 mV) than fluorene-grown cells (,51.6 ± 4.9 mV), but much more negatively charged than glucose-grown cells (,26.8 ± 3.3 mV), suggesting that the PAH substrate induced modifications on the physical properties of bacterial surfaces. Furthermore, protein-to-exopolysaccharide ratios detected during bacterial growth on phenanthrene were typical of biofilms developed under physicochemical stress conditions, caused by the presence of sparingly water-soluble chemicals as the sole carbon and energy source for growth, the maximum value for TP/EPS during growth on phenanthrene (1.9) being lower than the one obtained with fluorene (5.5). Finally, confocal laser microscopy observations using a gfp -labeled derivative strain revealed that, in the presence of phenanthrene, P. putida::gfp cells formed a biofilm on accessible crystal surfaces, whereas in the presence of fluorene the strain grew randomly between the crystal clusters. The results showed that P. putida was able to overcome the lower aqueous solubility of phenanthrene by adhering to the solid PAH throughout the production of extracellular polymeric substances, thus promoting the availability and uptake of such a hydrophobic compound. © 2005 Wiley Periodicals, Inc. [source] In silico genome-scale metabolic analysis of Pseudomonas putida KT2440 for polyhydroxyalkanoate synthesis, degradation of aromatics and anaerobic survivalBIOTECHNOLOGY JOURNAL, Issue 7 2010Seung Bum Sohn Abstract Genome-scale metabolic models have been appearing with increasing frequency and have been employed in a wide range of biotechnological applications as well as in biological studies. With the metabolic model as a platform, engineering strategies have become more systematic and focused, unlike the random shotgun approach used in the past. Here we present the genome-scale metabolic model of the versatile Gram-negative bacterium Pseudomonas putida, which has gained widespread interest for various biotechnological applications. With the construction of the genome-scale metabolic model of P. putida KT2440, PpuMBEL1071, we investigated various characteristics of P. putida, such as its capacity for synthesizing polyhydroxyalkanoates (PHA) and degrading aromatics. Although P. putida has been characterized as a strict aerobic bacterium, the physiological characteristics required to achieve anaerobic survival were investigated. Through analysis of PpuMBEL1071, extended survival of P. putida under anaerobic stress was achieved by introducing the ackA gene from Pseudomonas aeruginosa and Escherichia coli. [source] Rationally engineered biotransformation of p -nitrophenolBIOTECHNOLOGY PROGRESS, Issue 3 2010Matthew de la Peña Mattozzi Abstract An operon encoding enzymes responsible for degradation of the EPA priority contaminant para -nitrophenol (PNP) from Pseudomonas sp. ENV2030 contains more genes than would appear to be necessary to mineralize PNP. To determine some necessary genes for PNP degradation, the genes encoding the proposed enzymes in the degradation pathway (pnpADEC) were assembled into a broad-host-range, BioBricks-compatible vector under the control of a constitutive promoter. These were introduced into Escherichia coli DH10b and two Pseudomonas putida strains, one with a knockout of the aromatic transport TtgB and the parent with the native transporter. The engineered strains were assayed for PNP removal. E. coli DH10b harboring several versions of the refactored pathway was able to remove PNP from the medium up to a concentration of 0.2 mM; above which PNP was toxic to E. coli. A strain of P. putida harboring the PNP pathway genes was capable of removing PNP from the medium up to 0.5 mM. When P. putida harboring the native PNP degradation cluster was exposed to PNP, pnpADEC were induced, and the resulting production of ,-ketoadipate from PNP induced expression of its chromosomal degradation pathway (pcaIJF). In contrast, pnpADEC were expressed constitutively from the refactored constructs because none of the regulatory genes found in the native PNP degradation cluster were included. Although P. putida harboring the refactored construct was incapable of growing exclusively on PNP as a carbon source, evidence that the engineered pathway was functional was demonstrated by the induced expression of chromosomal pcaIJF. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [source] Factors Mediating Activity, Selectivity, and Substrate Specificity for the Thiamin Diphosphate-Dependent Enzymes Benzaldehyde Lyase and Benzoylformate DecarboxylaseCHEMBIOCHEM, Issue 12 2006Michael Knoll Abstract Benzaldehyde lyase from Pseudomonas fluorescens and benzoylformate decarboxylase from Pseudomonas putida are homologous thiamin diphosphate-dependent enzymes that catalyze carboligase and carbolyase reactions. Both enzymes catalyze the formation of chiral 2-hydroxy ketones from aldehydes. However, the reverse reaction has only been observed with benzaldehyde lyase. Whereas benzaldehyde lyase is strictly R specific, the stereoselectivity of benzoylformate decarboxylase from P. putida is dependent on the structure and orientation of the substrate aldehydes. In this study, the binding sites of both enzymes were investigated by using molecular modelling studies to explain the experimentally observed differences in the activity, stereo- and enantioselectivity and substrate specificity of both enzymes. We designed a detailed illustration that describes the shape of the binding site of both enzymes and sufficiently explains the experimental effects observed with the wild-type enzymes and different variants. These findings demonstrate that steric reasons are predominantly responsible for the differences observed in the (R)-benzoin cleavage and in the formation of chiral 2-hydroxy ketones. [source] | |||||||||||||