Habitabilidade em exoplanetas do tipo terrestre

A astrobiologia, ciência que estuda a origem, evolução, distribuição e futuro da vida, tem como escopo a procura por vida em outros ambientes, como por exemplo, exoplanetas, planetas estes localizados fora do sistema solar. Através de observações e modelos teóricos que estimam alguns parâmetros gerais (composição, tipos de compostos presentes) pode-se criar um ambiente possível para o exoplaneta. O ambiente em que o planeta está inserido é um fator que pode ser determinante em relação a sua habitabilidade. A Zona Habitável é um dos parâmetros passíveis de delimitar os planetas que poderiam ser habitáveis. Muitos planetas identificados até agora têm características similares à da Terra. Entretanto, a maioria dos planetas, com possibilidade de ser do tipo terrestre, identificados até hoje são considerados Super Terras Quentes (STQ), pois estão fora da zona habitável uma vez que estão muito próximos de suas estrelas (período orbital da ordem de dias ou menos). Alguns desses estariam presentes na Zona habitável de suas respectivas estrelas, como é o caso de Gl 581 g, um planeta do sistema multi-planetário da estrela Gl 581. A proximidade da estrela sugere que eles apresentam uma rotação afetada por forças de maré. Deste modo, a taxa rotacional do planeta é sincronizada, sendo que uma de suas faces não recebe a energia da estrela, tornando-se gelada e a outra face recebe constantemente a energia da estrela, tornando-se muito quente. Embora em geral descarta-se a habitabilidade em STQ, estudos de resistência de vida em ambientes extremos não são muito explorados. As características analisadas partirão da premissa que o tipo de vida procurada será semelhante ao tipo de vida que encontramos na Terra, isto é, utilizando a água como solvente e apresentando uma química baseada no elemento carbono (Kaltenegger, L. et al., 2008)... (Resumo completo, clicar acesso eletrônico abaixo)

Role of the denitrifying Haloarchaea in the treatment of nitrite-brines

Haloferax mediterranei is a denitrifying halophilic archaeon able to reduce nitrate and nitrite under oxic and anoxic conditions. In the presence of oxygen, nitrate and nitrite are used as nitrogen sources for growth. Under oxygen scarcity,this haloarchaeon uses both ions as electron acceptors via a denitrification pathway. In the present work, the maximal nitriteconcentration tolerated by this organism was determined by studying the growth of H. mediterranei in minimal medium containing30, 40 and 50 mM nitrite as sole nitrogen source and under initial oxic conditions at 42 °C. The results showed theability of H. mediterranei to withstand nitrite concentrations up to 50 mM. At the beginning of the incubation, nitrate wasdetected in the medium, probably due to the spontaneous oxidation of nitrite under the initial oxic conditions. The completeremoval of nitrite and nitrate was accomplished in most of the tested conditions, except in culture medium containing 50 mMnitrite, suggesting that this concentration compromised the denitrification capacity of the cells. Nitrite and nitrate reductases activities were analyzed at different growth stages of H. mediterranei. In all cases, the activities of the respiratory enzymeswere higher than their assimilative counterparts; this was especially the case for NirK. The denitrifying and possibly detoxifyingrole of this enzyme might explain the high nitrite tolerance of H. mediterranei. This archaeon was also able to remove60 % of the nitrate and 75 % of the nitrite initially present in brine samples collected from a wastewater treatment facility.These results suggest that H. mediterranei, and probably other halophilic denitrifying Archaea, are suitable candidates for thebioremediation of brines with high nitrite and nitrate concentrations.

Biodiversity of Archaea and floral of two inland saltern ecosystems in the Alto Vinalopó Valley, Spain

Background. The extraction of salt from seawater by means of coastal solar salterns is a very well-described process. Moreover, the characterization of these environments from ecological, biochemical and microbiological perspectives has become a key focus for many research groups all over the world over the last 20 years. In countries such as Spain, there are several examples of coastal solar salterns (mainly on the Mediterranean coast) and inland solar salterns, from which sodium chloride is obtained for human consumption. However, studies focused on the characterization of inland solar salterns are scarce and both the archaeal diversity and the plant communities inhabiting these environments remain poorly described. Results. Two of the inland solar salterns (termed Redonda and Penalva), located in the Alto Vinalopó Valley (Alicante, Spain), were characterized regarding their geological and physico-chemical characteristics and their archaeal and botanical biodiversity. A preliminary eukaryotic diversity survey was also performed using saline water. The chemical characterization of the brine has revealed that the salted groundwater extracted to fill these inland solar salterns is thalassohaline. The plant communities living in this environment are dominated by Sarcocornia fruticosa (L.) A.J. Scott, Arthrocnemum macrostachyum (Moris) K. Koch, Suaeda vera Forsk. ex Gmelin (Amaranthaceae) and several species of Limonium (Mill) and Tamarix (L). Archaeal diversity was analyzed and compared by polymerase chain reaction (PCR)-based molecular phylogenetic techniques. Most of the sequences recovered from environmental DNA samples are affiliated with haloarchaeal genera such as Haloarcula, Halorubrum, Haloquadratum and Halobacterium, and with an unclassified member of the Halobacteriaceae. The eukaryote Dunaliella was also present in the samples. Conclusions. To our knowledge, this study constitutes the first analysis centered on inland solar salterns located in the southeastern region of Spain. The results obtained revealed that the salt deposits of this region have marine origins. Plant communities typical of salt marshes are present in this ecosystem and members of the Halobacteriaceae family can be easily detected in the microbial populations of these habitats. Possible origins of the haloarchaea detected in this study are discussed.

Nitrogen metabolism in haloarchaea

The nitrogen cycle (N-cycle), principally supported by prokaryotes, involves different redox reactions mainly focused on assimilatory purposes or respiratory processes for energy conservation. As the N-cycle has important environmental implications, this biogeochemical cycle has become a major research topic during the last few years. However, although N-cycle metabolic pathways have been studied extensively in Bacteria or Eukarya, relatively little is known in the Archaea. Halophilic Archaea are the predominant microorganisms in hot and hypersaline environments such as salted lakes, hot springs or salted ponds. Consequently, the denitrifying haloarchaea that sustain the nitrogen cycle under these conditions have emerged as an important target for research aimed at understanding microbial life in these extreme environments. The haloarchaeon Haloferax mediterranei was isolated 20 years ago from Santa Pola salted ponds (Alicante, Spain). It was described as a denitrifier and it is also able to grow using NO3-, NO2- or NH4+ as inorganic nitrogen sources. This review summarizes the advances that have been made in understanding the N-cycle in halophilic archaea using Hfx mediterranei as a haloarchaeal model. The results obtained show that this microorganism could be very attractive for bioremediation applications in those areas where high salt, nitrate and nitrite concentrations are found in ground waters and soils.

Nitrogen regulation of protein–protein interactions and transcript levels of GlnK PII regulator and AmtB ammonium transporter homologs in Archaea

Gene homologs of GlnK PII regulators and AmtB-type ammonium transporters are often paired on prokaryotic genomes, suggesting these proteins share an ancient functional relationship. Here, we demonstrate for the first time in Archaea that GlnK associates with AmtB in membrane fractions after ammonium shock, thus, providing a further insight into GlnK-AmtB as an ancient nitrogen sensor pair. For this work, Haloferax mediterranei was advanced for study through the generation of a pyrE2-based counterselection system that was used for targeted gene deletion and expression of Flag-tagged proteins from their native promoters. AmtB1-Flag was detected in membrane fractions of cells grown on nitrate and was found to coimmunoprecipitate with GlnK after ammonium shock. Thus, in analogy to bacteria, the archaeal GlnK PII may block the AmtB1 ammonium transporter under nitrogen-rich conditions. In addition to this regulated protein–protein interaction, the archaeal amtB-glnK gene pairs were found to be highly regulated by nitrogen availability with transcript levels high under conditions of nitrogen limitation and low during nitrogen excess. While transcript levels of glnK-amtB are similarly regulated by nitrogen availability in bacteria, transcriptional regulators of the bacterial glnK promoter including activation by the two-component signal transduction proteins NtrC (GlnG, NRI) and NtrB (GlnL, NRII) and sigma factor σN (σ54) are not conserved in archaea suggesting a novel mechanism of transcriptional control.

Ferredoxin-dependent glutamate synthase: involvement in ammonium assimilation in Haloferax mediterranei

Glutamate synthase (GOGAT) is one of the two important enzymes involved in the ammonium assimilation pathway glutamine synthetase (GS)/GOGAT, which enables Hfx. mediterranei to thrive in media with low ammonium concentration or containing just nitrate as single nitrogen source. The gene coding for this enzyme, gltS, has been sequenced, analysed and compared with other GOGATs from different organisms from the three domains of life. According to its amino acid sequence, Hfx. mediterranei GOGAT displays high homology with those from other archaeal halophilic organisms and with the bacterial alpha-like subunit. Hfx. mediterranei GOGAT and GS expression was induced under conditions of ammonium restriction. The GOGAT protein was found to be a monomer with a molecular mass of 163.78 kDa, which is consistent with that estimated by gel filtration, 198 ± 30 kDa. The enzyme is highly ferredoxin dependent: activity was only observed with one of the two different 2Fe–2S ferredoxins chromatographically isolated from Hfx. mediterranei. The enzyme also displayed typical halophilic behaviour, being fully stable, and producing maximal activity, at salt concentrations from 3 to 4 M NaCl, pH 7.5 and a temperature of 50 °C.

Effects of nitrogen sources on the nitrate assimilation in Haloferax mediterranei: growth kinetics and transcriptomic analysis

The haloarchaeon Haloferax mediterranei is able to grow in a defined culture media not only in the presence of inorganic nitrogen salt but also with amino acid as the sole nitrogen source. Assimilatory nitrate and nitrite reductases, respectively, catalyze the first and second reactions. The genes involved in this process are nasA, which encodes nitrate reductase and is found within the operon nasABC, and nasD, which encodes nitrite reductase. These genes are subjected to transcriptional regulation, being repressed in the presence of ammonium and induced with either nitrate or nitrite. This type of regulation has also been described when the amino acids are used as nitrogen source in the minimal media. Furthermore, it has been observed that the microorganism growth depends on nitrogen source, obtaining the lowest growth rate in the presence of nitrate and aspartate. In this paper, we present the results of a comparative study of microorganism growth and transcriptomic analysis of the operon nasABC and gene nasD in different nitrogen sources. The results are the first ever produced in relation to amino acids as nitrogen sources within the Halobacteriaceae family.

Cu-NirK from Haloferax mediterranei as an example of metalloprotein maturation and exportation via Tat system

The green Cu-NirK from Haloferax mediterranei (Cu-NirK) has been expressed, refolded and retrieved as a trimeric enzyme using an expression method developed for halophilic Archaea. This method utilizes Haloferax volcanii as a halophilic host and an expression vector with a constitutive and strong promoter. The enzymatic activity of recombinant Cu-NirK was detected in both cellular fractions (cytoplasmic fraction and membranes) and in the culture media. The characterization of the enzyme isolated from the cytoplasmic fraction as well as the culture media revealed important differences in the primary structure of both forms indicating that Hfx. mediterranei could carry out a maturation and exportation process within the cell before the protein is exported to the S-layer. Several conserved signals found in Cu-NirK from Hfx. mediterranei sequence indicate that these processes are closely related to the Tat system. Furthermore, the N-terminal sequence of the two Cu-NirK subunits constituting different isoforms revealed that translation of this protein could begin at two different points, identifying two possible start codons. The hypothesis proposed in this work for halophilic Cu-NirK processing and exportation via the Tat system represents the first approximation of this mechanism in the Halobacteriaceae family and in Prokarya in general.

Regulation of ammonium assimilation in Haloferax mediterranei: Interaction between glutamine synthetase and two GlnK proteins

GlnK proteins belong to the PII superfamily of signal transduction proteins and are involved in the regulation of nitrogen metabolism. These proteins are normally encoded in an operon together with the structural gene for the ammonium transporter AmtB. Haloferax mediterranei possesses two genes encoding for GlnK, specifically, glnK1 and glnK2. The present study marks the first investigation of PII proteins in haloarchaea, and provides evidence for the direct interaction between glutamine synthetase and both GlnK1 and GlnK2. Complex formation between glutamine synthetase and the two GlnK proteins is demonstrated with pure recombinant protein samples using in vitro activity assays, gel filtration chromatography and western blotting. This protein–protein interaction increases glutamine synthetase activity in the presence of 2-oxoglutarate. Separate experiments that were carried out with GlnK1 and GlnK2 produced equivalent results.

Haloferax mediterranei GlnK proteins are post-translationally modified by uridylylation

In this work we report for the first time a post-translational modification of PII homologues from the Archaea Domain. Haloferax mediterranei is the first haloarchaea whose PII proteins have been studied, it possesses two of them (GlnK1 and GlnK2), both encoded adjacent to a gene for the ammonia transporter Amt. An approach based on 2DE, anti-GlnK immunoblot and peptide mass fingerprint (MALDI-TOF-MS) of the reactive spots showed that GlnK proteins in H. mediterranei are post-translationally uridylylated. A third spot with lower pI suggests the existence of a non-descript post-translational modification in this protein family.

Evaluación del trabajo colaborativo en Iniciación a la Investigación en Biología

Tras el trabajo previo de diseño de la asignatura Iniciación a la Investigación en Biología, el equipo multidisciplinar de profesores y profesoras de la Red Docente INVES ha desarrollado una metodología propia de trabajo en equipo, no sólo entre el diferente profesorado que la compone, sino también con el profesorado de la asignatura Estadística, con la que se comparten objetivos de aprendizaje comunes. Se ha optimizado el sistema de evaluación del trabajo colaborativo del alumnado, mediante el uso de rubricas y auto-evaluación. Dicho trabajo consiste en el diseño y desarrollo de un proyecto de investigación bibliométrico de temática biológica realizado por los estudiantes, propiciando la adquisición de competencias transversales mediante una dinámica de trabajo en grupo que culmina en la edición de unas Jornadas Científicas. Por otra parte, se han consensuado criterios comunes de evaluación continua, mejorando en la eficiencia de la evaluación, y determinado un incremento de la capacidad de aprendizaje del alumnado a lo largo de los cursos 2010-11 al 2013-14. La oferta formativa se completa mediante la formación de un grupo de Alto Rendimiento Académico con docencia en lengua inglesa. Esto permite al alumnado implementar el objetivo general de compresión de lengua extranjera inglés en lo relativo al ámbito científico.

English as the medium of instruction: a response to internationalization

The status of English as a lingua franca has led European universities to implement the use of this language as a medium of instruction (EMI). This study presents an analysis of the status quo of EMI at the University of Alicante. It takes into account the institution`s language policy and the programs which offer subjects in English, as well as the challenges, needs and benefits of the professors and students. Qualitative and quantitative data was collected by means of questionnaires and semi-structured interviews. The findings of this needs analysis will help us to create an action plan that will include teacher and student training schemes to foster internationalization.

Seguimiento de la implantación del Grado en Biología. Estrategias para la coordinación docente

Durante el curso académico 2013-2014 ha tenido lugar la implantación del último curso del Grado en Biología. Esto permite hacer un análisis completo de la implantación del título así como un estudio sobre el diseño y desarrollo del Trabajo de fin de Grado y la adaptación del Programa de Prácticas externas. Estrategias de trabajo a través de las comisiones de semestre, comisión de garantía de calidad del Centro, reuniones con el alumnado, grupos de acción tutorial, etc. han permitido detectar anomalías y reforzar aspectos positivos para el correcto funcionamiento del título. Cabe resaltar la alta participación del alumnado en la asignatura optativa de prácticas externas, así como un número elevado de alumnos/as matriculados en las optativas de cuarto curso vinculadas al Itinerario 2: Biotecnología y Biosanitaria. En cuanto a los Trabajos de Fin de Grado en curso, destaca el elevado número de trabajos experimentales, seguidos de los bibliográficos y en menor medida, los vinculados a empresas/instituciones.