Departamento de Biología Animal, Biología Vegetal y Ecología
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Ítem Depósito captador de lluvia para riego con efecto protector tipo planta nodriza(OEMP, 2011-11-29) Muñoz-Rodríguez, Juan José; García-Fuentes, AntonioDepósito captador de lluvia para riego con efecto protector tipo nodriza, que comprende una estructura cilíndrica con malla de sombreado y un depósito de agua (D), con una tapadera cóncava (T), con una válvula antirretorno que permite la entrada por gravedad del agua de lluvia, conectado dicho depósito a un tubo de riego por goteo, que puede estar controlado por un temporizador o un dispositivo de descarga retardada, con posibilidad de instalación de un mecanismo fotosensible. El depósito puede tener una prolongación a modo de visera hecha de material microperforado, y puede estar conectado en serie con otros depósitos análogos. El dispositivo planteado se compone de dos componentes. El primer componente es un depósito capaz de almacenar agua, recoger agua procedente de la precipitación evitando su pérdida y suministrarla a una planta. El suministro de agua se realiza paulatinamente, mediante un mecanismo retardado tras las precipitaciones, de tal forma que además de aumentar la cantidad de agua suministrada a la planta, aumenta el periodo de tiempo en el que el agua está disponible para la planta, manteniendo el suelo húmedo y aumentando el balance de número de días de lluvia/días secos. Así la planta recibe un aporte hídrico no sólo en los días de lluvia, sino también en los días posteriores, procedente del agua recogida en el depósito. El segundo componente es una estructura capaz de suministrar sombra y protección mecánica a una planta y, de este modo, reducir el exceso de radiación solar, la acción perjudicial de inclemencias meteorológicas y/o animales herbívoros. Esta estructura está diseñada para no reducir el aporte hídrico natural que recibiría la planta.Ítem A novel TctA citrate transporter from an activated sludge metagenome: Structural and mechanistic predictions for the TTT family(2014) Batista-García, Ramón; Sánchez-Reyes, Ayixon; Millán, César; González-Zuñiga, Víctor Manuel; Folch, Jorge Luis; Juárez-Ramírez, SoledadWe isolated a putative citrate transporter of the tripartite tricarboxylate transporter (TTT) class from a metagenomic library of activated sludge from a sewage treatment plant. The transporter, dubbed TctA_ar, shares ~50% sequence identity with TctA of Comamonas testosteroni (TctA_ct) and other b-Proteobacteria, and contains two 20-amino acid repeat signature sequences, considered a hallmark of this particular transporter class. The structures for both TctA_ar and TctA_ct were modeled with I-TASSER and two possible structures for this transporter family were proposed. Docking assays with citrate resulted in the corresponding sets of proposed critical residues for function. These models suggest functions for the 20-amino acid repeats in the context of the two different architectures. This constitutes the first attempt at structure modeling of the TTT family, to the best of our knowledge, and could aid functional understanding of this little-studied family.Ítem Characterization of Lignocellulolytic Activities from a Moderate Halophile Strain of Aspergillus caesiellus Isolated from a Sugarcane Bagasse Fermentation(PLOS ONE, 2014-08-27) Batista-García, Ramón; Balcázar, Edgar; Miranda, Estefan; Sánchez-Reyes, Ayixon; Cuervo, Laura; Aceves, Denise; Atriztán, Karina; Morales-Herrera, Catalina; Rodríguez-Hernández, Rocío; Folch, Jorge LuisA moderate halophile and thermotolerant fungal strain was isolated from a sugarcane bagasse fermentation in the presence of 2 M NaCl that was set in the laboratory. This strain was identified by polyphasic criteria as Aspergillus caesiellus. The fungus showed an optimal growth rate in media containing 1 M NaCl at 28°C and could grow in media added with up to 2 M NaCl. This strain was able to grow at 37 and 42°C, with or without NaCl. A. caesiellus H1 produced cellulases, xylanases, manganese peroxidase (MnP) and esterases. No laccase activity was detected in the conditions we tested. The cellulase activity was thermostable, halostable, and no differential expression of cellulases was observed in media with different salt concentrations. However, differential band patterns for cellulase and xylanase activities were detected in zymograms when the fungus was grown in different lignocellulosic substrates such as wheat straw, maize stover, agave fibres, sugarcane bagasse and sawdust. Optimal temperature and pH were similar to other cellulases previously described. These results support the potential of this fungus to degrade lignocellulosic materials and its possible use in biotechnological applications.Ítem A Novel Expansin Protein from the White-Rot Fungus Schizophyllum commune(PLOS, 2015) Batista-García, Ramón; Tovar-Herrera, Omar; Sánchez-Carbente, María; Iracheta, María Magdalena; Arévalo-Niño, Katiushka; Folch, Jorge LuisA novel expansin protein (ScExlx1) was found, cloned and expressed from the Basidiomycete fungus Schizophylum commune. This protein showed the canonical features of plant expansins. ScExlx1 showed the ability to form “bubbles” in cotton fibers, reduce the size of avicel particles and enhance reducing sugar liberation from cotton fibers pretreated with the protein and then treated with cellulases. ScExlx1 was able to bind cellulose, birchwood xylan and chitin and this property was not affected by different sodium chloride concentrations. A novel property of ScExlx1 is its capacity to enhance reducing sugars (N-acetyl glucosamine) liberation from pretreated chitin and further added with chitinase, which has not been reported for any expansin or expansin-like protein. To the best of our knowledge, this is the first report of a bona fide fungal expansin found in a basidiomycete and we could express the bioactive protein in Pichia pastoris.Ítem Identification of a novel carbohydrate esterase from Bjerkandera adusta: Structural and function predictions through bioinformatics analysis and molecular modeling(Wiley, 2015-01) Batista-García, Ramón; Valdés, Gilberto; Cuervo, Laura; Sánchez-Carbente, María; Balcázar, Edgar; Lira, Verónica; Folch, Jorge LuisA new gene from Bjerkandera adusta strain UAMH 8258 encoding a carbohydrate esterase (designated as BacesI) was isolated and expressed in Pichia pastoris. The gene had an open reading frame of 1410 bp encoding a polypeptide of 470 amino acid residues, the first 18 serving as a secretion signal peptide. Homology and phylogenetic analyses showed that BaCesI belongs to carbohydrate esterases family 4. Three-dimensional modeling of the protein and normal mode analysis revealed a breathing mode of the active site that could be relevant for esterase activity. Furthermore, the overall negative electrostatic potential of this enzyme suggests that it degrades neutral substrates and will not act on negative substrates such as peptidoglycan or p-nitrophenol derivatives. The enzyme shows a specific activity of 1.118 U mg21 protein on 2-naphthyl acetate. No activity was detected on p-nitrophenol derivatives as proposed from the electrostatic potential data. The deacetylation activity of the recombinant BaCesI was confirmed by measuring the release of acetic acid from several substrates, including oat xylan, shrimp shell chitin, N-acetylglucosamine, and natural substrates such as sugar cane bagasse and grass. This makes the protein very interesting for the biofuels production industry from lignocellulosic materials and for the production of chitosan from chitin.Ítem From lignocellulosic metagenomes to lignocellulolytic genes: trends, challenges and future prospects(Wiley, 2016) Batista-García, Ramón; Sánchez-Carbente, María; Talia, Paola; Jackson, Stephen A.; O'Leary, Niall D.; Dobson, Alan D.W.; Folch, Jorge LuisLignocellulose is the most abundant biomass on Earth with immense potential to act as a primary resource for the production of a range of compounds currently obtained from fossil fuel sources. However, lignocellulosic feedstocks remain largely underexploited due to the complex mixture of recalcitrant polymers present, whose structural features hinder access to the utilizable monosaccharide reservoir within cellulose. Various fungi and bacteria have been identifi ed that can enzymatically decompose lignocellulose to its monomeric compounds for use as carbon sources. The investigation of such lignocellulolytic organisms has proven very useful in gaining primary insights into degradation processes and key microbial enzymes, but the established limitations of culture-based approaches suggest that we have yet to understand the full range of lignocellulolytic mechanisms, likely expressed within natural systems. In this review, we focus on metagenomic approaches to study lignocellulose degradation from structural and functional perspectives, which may provide novel insights into this process in order to rationally design methods for the extraction of compounds from biomass that could enhance biorefinery efficiencies.Ítem Xenobiotic Compounds Degradation by Heterologous Expression of a Trametes sanguineus Laccase in Trichoderma atroviride(PLOS, 2016) Batista-García, Ramón; Balcázar, Edgar; Méndez-Lorenzo, Luz Helena; Esquivel, Ulises; Savary, Oliver; Cabana, Hubert; Herrera-Estrella, Alfredo; Folch, Jorge Luis; Kumar, Vaidyanathan Vinoth; Ayala-Aceves, MarcelaFungal laccases are enzymes that have been studied because of their ability to decolorize and detoxify effluents; they are also used in paper bleaching, synthesis of polymers, bioremediation, etc. In this work we were able to express a laccase from Trametes (Pycnoporus) sanguineus in the filamentous fungus Trichoderma atroviride. For this purpose, a transformation vector was designed to integrate the gene of interest in an intergenic locus near the blu17 terminator region. Although monosporic selection was still necessary, stable integration at the desired locus was achieved. The native signal peptide from T. sanguineus laccase was successful to secrete the recombinant protein into the culture medium. The purified, heterologously expressed laccase maintained similar properties to those observed in the native enzyme (Km and kcat and kcat/km values for ABTS, thermostability, substrate range, pH optimum, etc). To determine the bioremediation potential of this modified strain, the laccase-overexpressing Trichoderma strain was used to remove xenobiotic compounds. Phenolic compounds present in industrial wastewater and bisphenol A (an endocrine disruptor) from the culture medium were more efficiently removed by this modified strain than with the wild type. In addition, the heterologously expressed laccase was able to decolorize different dyes as well as remove benzo[α]pyrene and phenanthrene in vitro, showing its potential for xenobiotic compound degradation.Ítem Simple screening protocol for identification of potential mycoremediation tools for the elimination of polycyclic aromatic hydrocarbons and phenols from hyperalkalophile industrial effluents(Elsevier, 2017) Batista-García, Ramón; Kumar, Vaidyanathan Vinoth; Ariste, Arielle; Tovar-Herrera, Omar; Savary, Olivier; Peidro-Guzmán, Heidy; González-Abradelo, Deborah; Jackson, Stephen A.; Dobson, Alan D.W.; Sánchez-Carbente, María; Cabana, Hubert; Folch, Jorge Luis; Leduc, RolandA number of fungal strains belonging to the ascomycota, basidiomycota and zygomycota genera were subjected to an in vitro screening regime to assess their ligninolytic activity potential, with a view to their potential use in mycoremediation based strategies to remove phenolic compounds and polycyclic aromatic hydrocarbons (PAHs) from industrial wastewaters. All six basidiomycetes completely decolorized remazol brilliant blue R (RBBR), while also testing positive in both the guaiacol and gallic acid tests indicating good levels of lignolytic activity. All the fungi were capable of tolerating phenanthrene, benzo-α- pyrene, phenol and p-chlorophenol in agar medium at levels of 10 ppm. Six of the fungal strains, Pseudogymnoascus sp., Aspergillus caesiellus, Trametes hirsuta IBB 450, Phanerochate chrysosporium ATCC 787, Pleurotus ostreatus MTCC 1804 and Cadophora sp. produced both laccase and Mn peroxidase activity in the ranges of 200–560 U/L and 6–152 U/L, respectively, in liquid media under nitrogen limiting conditions. The levels of adsorption of the phenolic and PAHs were negligible with 99% biodegradation being observed in the case of benzo-α-pyrene, phenol and p chlorophenol. The aforementioned six fungal strains were also found to be able to effectively treat highly alkaline industrial wastewater (pH 12.4). When this wastewater was supplemented with 0.1 mM glucose, all of the tested fungi, apart from A. caesiellus, displayed the capacity to remove both the phenolic and PAH compounds. Based on their biodegradative capacity we found T. hirsuta IBB 450 and Pseudogymnoascus sp., to have the greatest potential for further use in mycoremediation based strategies to treat wastestreams containing phenolics and PAHs.Ítem Safety of a proteoliposome from Neisseria meningitides as adjuvant for a house dust mite allergy vaccine(Taylor & Francis, 2017-07-12) Perez-Llano, YordanisThe proteoliposome (PL) of Neisseria meningitidis serogroup B has been reported as a safe and potent vaccine adjuvant, inducing a TH1-skewed response. The present study describes a pre-clinical safety evaluation of an allergy therapeutic vaccine candidate based on purified allergens from Dermatophagoides siboney house dust mite and PL as adjuvant, both components adsorbed onto aluminum hydroxide gel. Two separate studies of acute toxicity evaluation were performed in mice and rabbits, and two repeatdose studies were conducted in non-sensitized and allergen-sensitized Balb/c mice, respectively. The study in sensitized mice intends to model a therapeutic setting. Aerosolized allergen challenge was used in both settings to model natural respiratory exposure. In the therapeutic setting, mice were administered with three doses containing 2 lg allergen at weekly intervals [subcutaneous route] and subsequently challenged with aerosolized allergen for 6 consecutive days. Parameters of general toxicity effects were assessed via measures of behavior, body weight, food and water consumption, and macroscopic evaluation of organs. Histological examination of organs and the injection site was performed. Potential immunotoxicity effects at the systemic level were assessed by blood eosinophil counting and serum allergen specific IgE by ELISA The vaccine did not produce general or functional toxic effects of significance, at a dose up to 100 lg allergen per kg body weight. An expected local reaction at the injection site was observed, which could be attributed mostly to the immunological effect of aluminum hydroxide. The models implemented here suggest an acceptable safety profile of this vaccine for testing in clinical trials of allergy immunotherapy.Ítem Schizophyllum commune: An unexploited source for lignocellulose degrading enzymes(Wiley, 2018) Perez-Llano, YordanisÍtem Omics Approaches: Impact on Bioremediation Techniques(Springer, 2018) Perez-Llano, YordanisÍtem First demonstration that ascomycetous halophilic fungi (Aspergillus sydowii T and Aspergillus destruens) are useful in xenobiotic mycoremediation under high salinity conditions(Elsevier, 2019) Batista-García, Ramón; González-Abradeloa, Deborah; Pérez-Llano, Yordanis; Peidro-Guzmán, Heidy; Sánchez-Carbente, María; Folch, Jorge Luis; Aranda-Ballesteros, Elizabet; Vaidyanathan, Vinoth Kumar; Cabana, Hubert; Gunde-Cimerman, NinaPolycyclic aromatic hydrocarbons (PAH) and pharmaceutical compounds (PhC) are xenobiotics present in many saline wastewaters. Although fungi are known for their ability to remove xenobiotics, the potential of halophilic fungi to degrade highly persistent pollutants was not yet investigated. The use of two halophilic fungi, Aspergillus sydowii and Aspergillus destruens, for the elimination of PAH and PhC at saline conditions was studied. In saline synthetic medium both fungi used benzo-α-pyrene and phenanthrene as sole carbon source and removed over 90% of both PAH, A. sydowii due to biodegradation and A. destruens to bioadsorption. They removed 100% of a mixture of fifteen PAH in saline biorefinery wastewater. Test using Cucumis sativus demonstrated that wastewater treated with the two fungi lowered considerably the phytotoxicity. This study is the first demonstration that ascomycetous halophilic fungi, in contrast to other fungi (and in particular basidiomycetes) can be used for mycotreatments under salinity conditions.Ítem First demonstration that ascomycetous halophilic fungi (Aspergillus sydowii and Aspergillus destruens) are useful in xenobiotic mycoremediation under high salinity conditions(Elsevier, 2019-02-02) Perez-Llano, YordanisÍtem Laccases from Extremophiles(Springer, 2020) Perez-Llano, YordanisÍtem Stress Reshapes the Physiological Response of Halophile Fungi to Salinity(MDPI, 2020) Batista-García, Ramón; Pérez-Llano, Yordanis; Rodríguez-Pupo, Eya Caridad; Druzhinina, Irina; Chenthamara, Komal; Cai, Feng; Gunde-Cimerman, Nina; Gostinčar, Cene; Kostanjšek, Rok; Folch, Jorge Luis; Sánchez-Carbente, MaríaBackground: Mechanisms of cellular and molecular adaptation of fungi to salinity have been commonly drawn from halotolerant strains and few studies in basidiomycete fungi. These studies have been conducted in settings where cells are subjected to stress, either hypo- or hyperosmotic, which can be a confounding factor in describing physiological mechanisms related to salinity. Methods: We have studied transcriptomic changes in Aspergillus sydowii, a halophilic species, when growing in three different salinity conditions (No NaCl, 0.5 M, and 2.0 M NaCl). Results: In this fungus, major physiological modifications occur under high salinity (2.0 M NaCl) and not when cultured under optimal conditions (0.5 M NaCl), suggesting that most of the mechanisms described for halophilic growth are a consequence of saline stress response and not an adaptation to saline conditions. Cell wall modifications occur exclusively at extreme salinity, with an increase in cell wall thickness and lamellar structure, which seem to involve a decrease in chitin content and an augmented content of alfa and beta-glucans. Additionally, three hydrophobin genes were differentially expressed under hypo- or hyperosmotic stress but not when the fungus grows optimally. Regarding compatible solutes, glycerol is the main compound accumulated in salt stress conditions, whereas trehalose is accumulated in the absence of salt. Conclusions: Physiological responses to salinity vary greatly between optimal and high salt concentrations and are not a simple graded effect as the salt concentration increases. Our results highlight the influence of stress in reshaping the response of extremophiles to environmental challenges.Ítem Haloadaptative Responses of Aspergillus sydowii to Extreme Water Deprivation: Morphology, Compatible Solutes, and Oxidative Stress at NaCl Saturation(MDPI, 2020) Batista-García, Ramón; Jiménez-Gómez, Irina; Valdés-Muñoz, Gisell; Moreno-Perlin, Tonatiuh; Mouriño-Pérez, Rosa Reyna; Sánchez-Carbente, María; Folch, Jorge Luis; Pérez-Llano, Yordanis; Gunde-Cimerman, Nina; Sánchez-Castellanos, NildaWater activity (aw) is critical for microbial growth, as it is severely restricted at aw < 0.90. Saturating NaCl concentrations (~5.0 M) induce extreme water deprivation (aw 0.75) and cellular stress responses. Halophilic fungi have cellular adaptations that enable osmotic balance and ionic/oxidative stress prevention to grow at high salinity. Here we studied the morphology, osmolyte synthesis, and oxidative stress defenses of the halophile Aspergillus sydowii EXF-12860 at 1.0 M and 5.13 M NaCl. Colony growth, pigmentation, exudate, and spore production were inhibited at NaCl-saturated media. Additionally, hyphae showed unpolarized growth, lower diameter, and increased septation, multicellularity and branching compared to optimal NaCl concentration. Trehalose, mannitol, arabitol, erythritol, and glycerol were produced in the presence of both 1.0 M and 5.13 M NaCl. Exposing A. sydowii cells to 5.13 M NaCl resulted in oxidative stress evidenced by an increase in antioxidant enzymes and lipid peroxidation biomarkers. Also, genes involved in cellular antioxidant defense systems were upregulated. This is the most comprehensive study that investigates the micromorphology and the adaptative cellular response of different non-enzymatic and enzymatic oxidative stress biomarkers in halophilic filamentous fungi.Ítem Stress reshapes the physiological response of halophile fungi to salinity(MDPI, 2020-02) Perez-Llano, YordanisÍtem Haloadaptative responses of Aspergillus sydowii to extreme water deprivation: morphology, compatible solutes, and oxidative stress at NaCl saturation(MDPI, 2020-11) Perez-Llano, YordanisÍtem Not Only Toxic but Repellent: What Can Organisms' Responses Tell Us about Contamination and What Are the Ecological Consequences When They Flee from an Environment?(MDPI, 2020-12) Araújo, Cristiano; Laissaoui, Abdelmourhit; Silva, Daniel C.V.R.; Ramos-Rodríguez, Eloísa; González-Ortegón, Enrique; Espíndola, Evaldo L.G.; Baldó, Francisco; Mena, Freylan; Parra-Anguita, Gema; Blasco, Julián; López-Doval, Julio; Sendra, Marta; Banni, Mohammed; Moreno-Garrido, IgnacioThe ability of aquatic organisms to sense the surrounding environment chemically and interpret such signals correctly is crucial for their ecological niche and survival. Although it is an oversimplification of the ecological interactions, we could consider that a significant part of the decisions taken by organisms are, to some extent, chemically driven. Accordingly, chemical contamination might interfere in the way organisms behave and interact with the environment. Just as any environmental factor, contamination can make a habitat less attractive or even unsuitable to accommodate life, conditioning to some degree the decision of organisms to stay in, or move from, an ecosystem. If we consider that contamination is not always spatially homogeneous and that many organisms can avoid it, the ability of contaminants to repel organisms should also be of concern. Thus, in this critical review, we have discussed the dual role of contamination: toxicity (disruption of the physiological and behavioral homeostasis) vs. repellency (contamination-driven changes in spatial distribution/habitat selection). The discussion is centered on methodologies (forced exposure against non-forced multi-compartmented exposure systems) and conceptual improvements (individual stress due to the toxic effects caused by a continuous exposure against contamination-driven spatial distribution). Finally, we propose an approach in which Stress and Landscape Ecology could be integrated with each other to improve our understanding of the threat contaminants represent to aquatic ecosystems.
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