A new green fluorescent protein‐based bacterial biosensor for analysing phenanthrene fluxes

Tecon, Robin; Wells, Mona; Meer, Jan Roelof van der

doi:10.1111/j.1462-2920.2005.00948.x

Cited by 49 publications

(38 citation statements)

References 40 publications

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“…It has been reported that the solubility of PHE in water is about 1.1-1.2 mg/l [5,6]. However, in the process of PHE biodegradation, the concentration of PHE is depleted with incubation time while more PHE may be soluble till the substrate is exhausted.…”

mentioning

confidence: 97%

Biodegradation of phenanthrene in an anaerobic batch reactor: Growth kinetics

Nasrollahzadeh¹,

Najafpour

Pazouki

et al. 2010

CI&CEQ

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The purpose of the present research was to demonstrate the ability of mixed consortia of microorganisms to degrade high concentrations of phenanthrene (PHE) as the sole carbon source. Batch experiments were carried out by the induction of mineral salt medium containing PHE to the seed culture and monitoring PHE biodegradation. The microbial propagation was conducted using PHE concentrations in the range of 20 to 100 mg/l. The microbial growth on PHE was defined based on Monod and modified Logistic rate models. The kinetic studies revealed that maximum specific growth rates (μ m ) for PHE concentrations of 20, 50 and 100 mg/l were 0.12, 0.23 and 0.035 h -1 , respectively. The doubling times for microbial population in PHE concentrations of 20, 50 and 100 mg/l were 13, 15 and 17.5 h, respectively. Also, maximum cell dry weight (x m ) of 54.23 mg/l was achieved, while the inhibition coefficient was 0.023 h -1 . It was observed that the experimental data were well represented by the proposed models. It was also found that the biodegradation of PHE was successfully performed by the isolated strains.

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mentioning

confidence: 97%

Biodegradation of phenanthrene in an anaerobic batch reactor: Growth kinetics

Nasrollahzadeh¹,

Najafpour

Pazouki

et al. 2010

CI&CEQ

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“…in the concentration range from 200 nM to 1.2 mM (Wei et al 2010), whereas BMB-PL was weakly induced by naphthalene, salicylate and pyruvate (data not shown). It was assumed that no induction would occur in the presence of other PAHs whose solubility in water is even lower than PHE (0.076 mg L -1 ) (Tecon et al 2006).…”

Section: Resultsmentioning

confidence: 99%

Specific detection of bioavailable phenanthrene and mercury by bacterium reporters in the red soil

Ze-Ling

Le-le

et al. 2013

Int. J. Environ. Sci. Technol.

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Genetically engineered Pseudomonas putida reporters (BMB-PL and BMB-ME), which, respectively, carried phnS-luxCDABE and merR-egfp cassette, were used to determine bioavailable phenanthrene and mercury. Over a spiked range of concentrations and aged for 6 days in red soil samples, the reporters were tested to determine the optimal assay conditions and the bioavailable phenanthrene (0-60 mg kg ) were evaluated by the signal of the relative fluorescent units and relative luminescence units. Single contamination was carried out and good correlations were obtained between signal strength and pollutant concentrations, whereas interference and bioavailability repression were observed in dual-contamination experiments. Other heavy metal ions at nanomolar level did not interfere with BMB-ME measurement while BMB-PL showed some response to other polycyclic aromatic hydrocarbons or their intermediate products during degradation. Comparing high-performance liquid chromatography methods with the bacterial reporters, both BMB-ME and BMB-PL appeared to have a detection limit (mercury \40 lg kg -1 ; phenanthrene \24 mg kg -1 ) similar to the instrumental analysis. Although physical parameters may affect the interaction of pollutants with bioreporter cells, advantages include the inherent biological relevance of the response, rapid response time, and potential for field deployment. Our results strongly suggest that the BMB-ME and BMB-PL bioreporters constitute an adaptable system for easily detecting the bioavailability of mercury and phenanthrene in the red soils.

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“…To run 20 parallel microcosms, obtain 100 ml of modified tryptone-yeast medium (mTY) 14 , about 150 ml of 21C minimal medium 16 (MM), 5 ml of minimal medium agar (MMA; 0.5%, w/v), three potato dextrose agar plates (PDA; 1.5%, w/v) for P. ultimum, four blank plates, one PDA plate containing 2 mg L -1 cycloheximide, four agarose plates (3%, w/v) containing 0.3 g ml -1 activated carbon and 40 mg of FLU in 2 mg portions. 2.…”

Section: Preparation Of Media and Culturesmentioning

confidence: 99%

“…In more detail, three-ring PAH fluorene, the mycelial organism Pythium ultimum and the bioreporter bacterium Burkholderia sartisoli RP037-mChe 13 were applied in the described microcosm setups. The bacterium B. sartisoli RP037-mChe was originally constructed to study phenanthrene fluxes to the cell 14 and expresses enhanced green fluorescent protein (eGFP)…”

Section: Introductionmentioning

confidence: 99%

A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems

Schamfuß

Neu

Harms

et al. 2014

JoVE

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Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is not an appropriate measure for its availability; bioavailability rather depends on the dynamic interplay of potential mass transfer (flux) of a compound to a microbial cell and the capacity of the latter to degrade the compound. In water-unsaturated parts of the soil, mycelia have been shown to overcome bioavailability limitations by actively transporting and mobilizing organic compounds over the range of centimeters. Whereas the extent of mycelia-based transport can be quantified easily by chemical means, verification of the contaminant-bioavailability to bacterial cells requires a biological method. Addressing this constraint, we chose the PAH fluorene (FLU) as a model compound and developed a water unsaturated model microcosm linking a spatially separated FLU point source and the FLU degrading bioreporter bacterium Burkholderia sartisoli RP037-mChe by a mycelial network of Pythium ultimum. Since the bioreporter expresses eGFP in response of the PAH flux to the cell, bacterial FLU exposure and degradation could be monitored directly in the microcosms via confocal laser scanning microscopy (CLSM). CLSM and image analyses revealed a significant increase of the eGFP expression in the presence of P. ultimum compared to controls without mycelia or FLU thus indicating FLU bioavailability to bacteria after mycelia-mediated transport. CLSM results were supported by chemical analyses in identical microcosms. The developed microcosm proved suitable to investigate contaminant bioavailability and to concomitantly visualize the involved bacteria-mycelial interactions. Video LinkThe video component of this article can be found at

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A new green fluorescent protein‐based bacterial biosensor for analysing phenanthrene fluxes

Cited by 49 publications

References 40 publications

Biodegradation of phenanthrene in an anaerobic batch reactor: Growth kinetics

Biodegradation of phenanthrene in an anaerobic batch reactor: Growth kinetics

Specific detection of bioavailable phenanthrene and mercury by bacterium reporters in the red soil

A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems

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