Pyoverdine (PvdI) is the major siderophore secreted by Pseudomonas aeruginosa PAOI in order to get access to iron. After being loaded with iron in the extracellular medium, PvdI is transported across the bacterial outer membrane by the transporter, FpvAI. We used the spectral properties of PvdI to show that in addition to Fe(3+), this siderophore also chelates, but with lower efficiencies, all the 16 metals used in our screening. Afterwards, FpvAI at the cell surface binds Ag(+), Al(3+), Cd(2+), Co(2+), Cu(2+), Fe(3+), Ga(3+), Hg(2+), Mn(2+), Ni(2+) or Zn(2+) in complex with PvdI. We used Inductively Coupled Plasma-Atomic Emission Spectrometry to monitor metal uptake in P. aeruginosa: TonB-dependent uptake, in the presence of PvdI, was only efficient for Fe(3+). Cu(2+), Ga(3+), Mn(2+) and Ni(2+) were also transported into the cell but with lower uptake rates. The presence of Al(3+), Cu(2+), Ga(3+), Mn(2+), Ni(2+) and Zn(2+) in the extracellular medium induced PvdI production in P. aeruginosa. All these data allow a better understanding of the behaviour of the PvdI uptake pathway in the presence of metals other than iron: FpvAI at the cell surface has broad metal specificity at the binding stage and it is highly selective for Fe(3+) only during the uptake process.
While diatoms are widely present in terms of diversity and abundance in nature, few species are currently used for biotechnologically applications. Most studies have focussed on intracellularly synthesised eicosapentaenoic acid (EPA), a polyunsaturated fatty acid (PUFA) used for pharmaceutical applications. Applications for other intracellular molecules, such as total lipids for biodiesel, amino acids for cosmetic, antibiotics and antiproliferative agents, are at the early stage of development. In addition, the active principle component must be identified amongst the many compounds of biotechnological interest. Biomass from diatom culture may be applied to: (1). aquaculture diets, due to the lipid- and amino-acid-rich cell contents of these microorganisms, and (2). the treatment of water contaminated by phosphorus and nitrogen in aquaculture effluent, or heavy metal (bioremediation). The most original application of microalgal biomass, and specifically diatoms, is the use of silicon derived from frustules in nanotechnology. The competitiveness of biotechnologically relevant products from diatoms will depend on their cost of production. Apart from EPA, which is less expensive when obtained from Phaeodactylum tricornutum than from cod liver, comparative economic studies of other diatom-derived products as well as optimisation of culture conditions are needed. Extraction of intracellular metabolites should be also optimised to reduce production costs, as has already been shown for EPA. Using cell immobilisation techniques, benthic diatoms can be cultivated more efficiently allowing new, biotechnologically relevant products to be investigated.
Biotechnological applications of diatoms are still in development. Further development at the industrial scale will depend on optimisation of the culture process with the aim of reducing costs. Because of the photoautotrophic status of the majority of diatoms, microalgal cultures suffer from the limitation of light diffusion, which requires the development of suitable photobioreactors. Thus, genetically engineered microalgae that may be cultivated in heterotrophic conditions present a new opportunity. Other limiting factors, such as nutrients (phosphate, nitrogen, silicon), pH, temperature, bioturbation and many more must be taken into account. Most of the time, metabolic stress conditions lead to an overproduction of the products of interest, with a decrease in biomass production as a consequence. Outdoor cultures in open ponds are usually devoted to aquaculture for the feeding of shrimps and bivalve molluscs (commercial production), while closed axenic indoor/outdoor photobioreactors are used for biotechnological compounds of homogeneous composition (still at the laboratory scale). In addition to the optimum culture conditions that have to be taken into account for photobioreactor design, the localisation of produced metabolites (intra- or extracellular) may also be taken into account when choosing the design. Microalgal cell immobilisation may be a suitable technique for application to benthic diatoms, which are usually sensitive to bioturbation and/or metabolites which may be overexpressed.
International audienceContamination caused by pesticides in agriculture is a source of environmental poor water quality in some of the European Union countries. Without treatment or targeted mitigation, this pollution is diffused in the environment. Pesticides and some metabolites are of increasing concern because of their potential impacts on the environment, wildlife and human health. Within the context of the European Union (EU) water framework directive context to promote low pesticide-input farming and best management practices, the EU LIFE project ArtWET assessed the efficiency of ecological bioengineering methods using different artificial wetland (AW) prototypes throughout Europe. We optimized physical and biological processes to mitigate agricultural nonpoint-source pesticide pollution in artificial wetland ecosystems. Mitigation solutions were implemented at full-scale demonstration and experimental sites. We tested various bioremediation methods at seven experimental sites. These sites involved (1) experimental prototypes, such as vegetated ditches, a forest microcosm and 12 wetland mesocosms, and (2) demonstration prototypes: vegetated ditches, three detention ponds enhanced with technology of constructed wetlands, an outdoor bioreactor and a biomassbed. This set up provides a variety of hydrologic conditions, with some systems permanently flooded and others temporarily flooded. It also allowed to study the processes both in field and controlled conditions. In order to compare the efficiency of the wetlands, mass balances at the inlet and outlet of the artificial wetland will be used, taking into account the partition of the studied compound in water, sediments, plants, and suspended solids. The literature background necessary to harmonize the interdisciplinary work is reviewed here and the theoretical framework regarding pesticide removal mechanisms in artificial wetland is discussed. The development and the implementation of innovative approaches concerning various water quality sampling strategies for pesticide load estimates during flood, specific biological endpoints, innovative bioprocess applied to herbicide and copper mitigation to enhance the pesticide retention time within the artificial wetland, fate and transport using a 2D mixed hybrid finite element model are introduced. These future results will be useful to optimize hydraulic functioning, e.g., pesticide resident time, and biogeochemical conditions, e.g., dissipation, inside the artificial wetlands. Hydraulic retention times are generally too low to allow an optimized adsorption on sediment and organic materials accumulated in artificial wetlands. Absorption by plants is not either effective. The control of the hydraulic design and the use of adsorbing materials can be useful to increase the pesticides residence time and the contact between pesticides and biocatalyzers. Pesticide fluxes can be reduced by 50-80% when hydraulic pathways in artificial wetlands are optimized by increasing ten times the retention time, by recirculation of w...
Partly because of the low bioavailability of metals, the soil cleaning-up using phytoremediation is usually time-consuming. In order to enhance the amount of metals at the plant's disposal, the soil bioaugmentation coupled together with phytoextraction is an emerging technology. In this preliminary work, two agricultural soils which mainly differed in their Cr, Hg and Pb contents (LC, low-contaminated soil; HC, high-contaminated soil) were bioaugmented in laboratory conditions by either bacterial (Bacillus subtilis, Pseudomonas aeruginosa, Pseudomonas fluorescens or Ralstonia metallidurans) or fungal inocula (Aspergillus niger or Penicillium simplicissimum) and incubated during three weeks. The LC soil pots bioaugmented with A. niger and P. aeruginosa contained higher concentrations of Cr (0.08 and 0.25 mg.kg −1 dw soil) and Pb (0.25 and 0.3 mg.kg −1 dw soil) in the exchangeable fraction F1 (extraction with MgCl 2 ) by comparison with the non-bioaugmented soil where neither Cr nor Pb was detected. Conversely, immobilization of Cr and Pb in the soil were observed with the other microorganisms. The soil bioaugmentation not only modified the metal speciation for the most easily extractable fractions but also modified the distribution of metals in the other fractions, to a lesser extent nevertheless. The difference in microbial concentrations between the bioaugmented or not HC soils reached up to 1.8 log units. Thus the microorganisms that we chose for the soil bioaugmentation were competitive towards the indigenous microflora. The PCA analysis showed close positive relationships between the microorganisms which potentially produced siderophores in the soil and the amount of Cr and Pb in the fraction F1.
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