Instituto Universitario Of Biotecnologia Of Asturias

Corvera de Asturias, Spain

Instituto Universitario Of Biotecnologia Of Asturias

Corvera de Asturias, Spain
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Fernandez R.,University of Oviedo | Fernandez R.,Instituto Universitario Of Biotecnologia Of Asturias | Bertrand A.,University of Oviedo | Bertrand A.,Instituto Universitario Of Biotecnologia Of Asturias | And 5 more authors.
Journal of Hazardous Materials | Year: 2013

Two clones of Dittrichia viscosa (L.) Greuter from contrasting populations, DV-A (metallicolous) and DV-W (non-metallicolous), were studied to compare Cd accumulation and tolerance. After 10 days of hydroponic culture with 0, 5, 10, and 15mgCdL-1, metal accumulation and plant growth were measured as well as other stress markers such as decrease in the content of photosynthetic pigments, lipid peroxidation, phenols, H2O2, and free proline. We also analyzed the activity of the antioxidant enzymes guaiacol and ascorbate peroxidases, catalase, superoxide dismutase, and glutathione reductase as well as their isoform patterns. Our results confirmed a high Cd tolerance and accumulation in both clones of D. viscosa, which suggests that these traits are constitutive in this species. However, when the Cd concentration in solution exceeded 10mgCdL-1, DV-A was more tolerant than DV-W. The physiological mechanisms involved in Cd tolerance also differed between them, although phenols and guaiacol peroxidase played an important role in both clones. The effective Cd detoxification of DV-A consisted mainly in a promoted ascorbate peroxidase activity and better efficiency of catalase and glutathione reductase enzymes. © 2012 Elsevier B.V.


Muller C.R.,RWTH Aachen | Muller C.R.,Instituto Universitario Of Biotecnologia Of Asturias | Lavandera I.,Instituto Universitario Of Biotecnologia Of Asturias | Gotor-Fernandez V.,Instituto Universitario Of Biotecnologia Of Asturias | And 2 more authors.
ChemCatChem | Year: 2015

Deep-eutectic solvents (DES) are cost-effective, nonhazardous solvents that may be used in biocatalysis as nonconventional media that enable biotransformations with industrially sound high substrate loadings. Based on promising prognoses, this paper explores successfully the use of whole cells overexpressing oxidoreductases as biocatalysts for the stereoselective reduction of ketones in different DES-aqueous-media solutions. Enzymes like Ralstoniasp. alcohol dehydrogenase (ADH) and horse liver ADH, overexpressed in Escherichiacoli, remain active at different DES proportions (for example, DES:buffer, 80:20 v/v) to enable nonconventional biotransformations. Furthermore, the enantiomeric excesses obtained for a broad range of aromatic substrates increase significantly if DES is added, which provides a useful tool for combining high substrate loadings and improved enzymatic selectivities while working with nonhazardous solvents. Different DES are successfully employed, which gives hints of the potential that these emerging solvents may have. DESirable biocatalysis: Several enzymes, like Ralstoniasp. alcohol dehydrognase (RasADH) overexpressed in Escherichiacoli, remain active for the stereoselective reduction of ketones in various deep-eutectic-solvents (DES)-buffer solutions. Furthermore, the ee values obtained for a broad range of aromatic substrates increase significantly if the DES mixture is added. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Fernandez R.,University of Oviedo | Fernandez R.,Instituto Universitario Of Biotecnologia Of Asturias | Fernandez-Fuego D.,University of Oviedo | Fernandez-Fuego D.,Instituto Universitario Of Biotecnologia Of Asturias | And 6 more authors.
Environmental Science and Pollution Research | Year: 2014

In this paper, we examined Cd accumulation and PC synthesis in two clones of Dittrichia viscosa, one with a metallicolous (DV-A) and the other with a non-metallicolous origin (DV-W). The clones were cultured in vitro with 0 and 10 mg Cd L-1 in both short-term treatments (up to 72 h) and over 10 days. We also examined the influence of the culture medium dilution and the PC-synthesis inhibitor, l-buthionine-sulfoximine (BSO), on these parameters. Similar Cd accumulation values were found in the two clones. No synthesis of new thiolic compounds was observed in Cd-treated plants cultured in vitro in Murashige and Skoog medium up to 72 h when compared to controls. Dilution of the culture medium affected PC production, increasing it in 1/2 MS and especially in 1/4 MS. Cd uptake did not increase in the same way, but still hyperaccumulation levels were exceeded in all Cd treatments. BSO addition increased the sensitivity of D. viscosa to Cd and diminished Cd accumulation. Nevertheless, a poor correlation between PCs and Cd accumulation capacity was observed since the highest Cd content did not correspond to the highest PC levels. All these results obtained suggest that PCs are important in Cd accumulation and detoxification in D. viscosa and also that other mechanisms might be involved in these traits. © 2013 Springer-Verlag Berlin Heidelberg.


Fernandez R.,University of Oviedo | Fernandez R.,Instituto Universitario Of Biotecnologia Of Asturias | Fernandez-Fuego D.,University of Oviedo | Fernandez-Fuego D.,Instituto Universitario Of Biotecnologia Of Asturias | And 4 more authors.
Plant Physiology and Biochemistry | Year: 2014

Dittrichia viscosa (L.) Greuter is plant species commonly found in degraded zones of Asturias (Spain), where it accumulates high levels of Cd, but the mechanisms involved in this response in non-model plants have not been elucidated. In this way, we analysed the fraction of the total Cd bound to the cell walls, the ultrastructural localization of this metal, and non-protein thiol and organic acid concentrations of two clones of D.viscosa: DV-A (from a metal-polluted soil) and DV-W (from a non-polluted area). After 10 days of hydroponic culture with Cd, fractionation and ultrastructural localisation studies showed that most of the Cd accumulated by D.viscosa was kept in the cell wall. The non-protein thiol content rose in D.viscosa with Cd exposure, especially in the non-metallicolous DV-W clone, and in both clones we found with Cd exposure a synthesis de novo of phytochelatins PC2 and PC3 in shoots and roots and also of other phytochelatin-related compounds, particularly in roots. Regarding organic acids, their concentration in both clones decreased in shoots after Cd treatment, but increased in roots, mainly due to changes in the citric acid concentration. Thus, retention of Cd in the cell wall seems to be the first strategy in response to metal entry in D.viscosa and once inside cells non-protein thiols and organic acids might also participate in Cd tolerance. © 2014.

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