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Putida F1 (putida + f1)
Kinds of Putida F1 Selected AbstractsDynamics of Benzene and Toluene Degradation in Pseudomonas putida F1 in the Presence of the Alternative Substrate SuccinateENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 4 2007I. Rüegg Abstract In batch and continuous culture, the regulation of benzene and toluene degradation by Pseudomonas putida,F1 was investigated in the presence of the alternative carbon and energy source succinate. In batch culture, benzene and toluene were used simultaneously, whereas succinate suppressed benzene consumption under carbon excess conditions resulting in diauxic growth. In carbon-limited continuous culture mixed substrate growth was observed. Since in nature low substrate concentrations and ever changing conditions prevail, this paper focuses on the dynamics of benzene/toluene degradation, biomass synthesis, and the regulation of benzene/toluene-degrading enzymes in cultures growing continuously at a dilution rate of 0.1,h,1, when shifting the supply of the carbon and energy source from succinate to various mixtures of succinate and benzene/toluene, or to benzene only. When the succinate concentration was kept constant (1.25,mM) and the medium was supplemented with benzene (2,mM), growth with benzene began already two hours after the shift. In contrast, replacing succinate with benzene only led to a wash out of biomass for more then ten hours, before biomass production from benzene started. A striking and reproducible transition pattern was observed for all shifts where the succinate concentration was reduced or succinate was omitted. After an initial period of biomass production from benzene, the culture collapsed and a wash-out of biomass was observed. However, this wash-out was not accompanied by an increase in benzene in the cultivation liquid, indicating a benzene uptake without conversion into biomass. Another possibility is that in phases of low biomass concentrations, cells were only able to use the low amounts of benzene/toluene dissolved in the cultivation liquid yielding low biomass concentrations whereas in phases of high biomass concentrations, they were able to rapidly utilize the aromatic solvents so that additional benzene from the gas phase diffused into the cultivation liquid resulting in more biomass production. In most cases, growth resumed again after 10 to 80,h. Currently, the reasons for the decrease in biomass after the first rise are unknown. However, several indications rule out intoxication of the cells by either the solvents benzene or toluene themselves, or by toxic degradation intermediates, or by-products. [source] Evolution of a chlorobenzene degradative pathway among bacteria in a contaminated groundwater mediated by a genomic island in RalstoniaENVIRONMENTAL MICROBIOLOGY, Issue 3 2003Tina Andrea Müller Summary The genetic structure of two Ralstonia spp., strain JS705 and strain JS745, isolated from the same groundwater aquifer, was characterized with respect to the degradation capacities for toluene and chlorobenzene degradation. Cosmid library construction, cloning, DNA sequencing and mating experiments indicated that the genes for chlorobenzene degradation in strain JS705 were a mosaic of the clc genes, previously described for Pseudomonas sp. strain B13, and a 5 kb fragment identical to strain JS745. The 5 kb fragment identical to both JS705 and JS745 was flanked in JS705 by one complete and one incomplete insertion (IS) element. This suggested involvement of the IS element in mobilizing the genes from JS745 to JS705, although insertional activity of the IS element in its present configuration could not be demonstrated. The complete genetic structure for chlorobenzene degradation in strain JS705 resided on a genomic island very similar to the clc element (Ravatn, R., Studer, S., Springael, D., Zehnder, A.J., van der Meer, J.R. 1998. Chromosomal integration, tandem amplification, and deamplification in Pseudomonas putida F1 of a 105-kilobase genetic element containing the chlorocatechol degradative genes from Pseudomonas sp. strain B13. J Bacteriol 180: 4360,4369). The unique reconstruction of formation of a metabolic pathway through the activity of IS elements and a genomic island in the chlorobenzene-degrading strain JS705 demonstrated how pathway evolution can occur under natural conditions in a few ,steps'. [source] Use of a whole-cell biosensor to assess the bioavailability enhancement of aromatic hydrocarbon compounds by nonionic surfactantsBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2008Angela Keane Abstract The whole-cell bioluminescent biosensor Pseudomonas putida F1G4 (PpF1G4), which contains a chromosomally-based sep-lux transcriptional fusion, was used as a tool for direct measurement of the bioavailability of hydrophobic organic compounds (HOCs) partitioned into surfactant micelles. The increased bioluminescent response of PpF1G4 in micellar solutions (up to 10 times the critical micellar concentration) of Triton X-100 and Brij 35 indicated higher intracellular concentrations of the test compounds, toluene, naphthalene, and phenanthrene, compared to control systems with no surfactants present. In contrast, Brij 30 caused a decrease in the bioluminescent response to the test compounds in single-solute systems, without adversely affecting cell growth. The decrease in bioluminescent response in the presence of Brij 30 did not occur in the presence of multiple HOCs extracted into the surfactant solutions from crude oil and creosote. The effect of the micellar solutions on the toluene biodegradation rate was consistent with the bioluminescent response in single-solute systems. None of the surfactants were toxic to PpF1G4 at the doses employed in this study, and PpF1G4 did not produce a bioluminescent response to the surfactants nor utilize them as growth substrates. TEM images suggest that the surfactants did not rupture the cell membranes. The results demonstrate that for Pseudomonas putida F1, nonionic surfactants such as Triton X-100 and Brij 35, at doses between 2 and 10 CMC, may increase the bioavailability and direct uptake of micellar phase HOCs that are common pollutants at contaminated sites. Biotechnol. Bioeng. 2008;99: 86,98. © 2007 Wiley Periodicals, Inc. [source] Purification, crystallization and preliminary X-ray diffraction studies of the three components of the toluene 2,3-dioxygenase enzyme systemACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 7 2005Kyoung Lee Pseudomonas putida F1 can grow with toluene as its sole source of carbon and energy. The initial reaction of the degradation of toluene is catalyzed by a three-component toluene dioxygenase enzyme system consisting of a reductase (ReductaseTOL), a ferredoxin (FerredoxinTOL) and a Rieske non-heme iron dioxygenase (OxygenaseTOL). The three components and the apoenzyme of the dioxygenase (apo-OxygenaseTOL) were overexpressed, purified and crystallized. ReductaseTOL diffracts to 1.8,Å and belongs to space group P41212, with unit-cell parameters a = b = 77.1, c = 156.3,Å. FerredoxinTOL diffracts to 1.2,Å and belongs to space group P21, with unit-cell parameters a = 30.5, b = 52.0, c = 30.95,Å, , = 113.7°. Apo-OxygenaseTOL and OxygenaseTOL diffract to 3.2,Å and belong to space group P4332, with unit-cell parameters a = 235.9,Å and a = 234.5,Å, respectively. [source] |