216 research outputs found
Novel biocatalysts by identification and design
Enzymes produced from bacteria and eukaryotic organisms are presently being used for a large variety of different biotechnological applications. The rapidly increasing demand for enzymes which are active towards novel and often non-natural substrates has triggered the development of novel molecular biological methods of enzyme isolation and design. The metagenome approach is a cultivation-independent method which allows the direct cloning and expression of environmental DNA thereby providing access to a wealth of so-far unknown biocatalysts. Additionally, newly identified or existing biocatalysts can be further optimized by different methods of directed evolution. Here, the principle of the metagenome approach is outlined and a strategy is presented for the optimization of a bacterial lipase using a combination of rational design and directed evolution.
Der Umzug der Menschheit: Die transformative Kraft der StÀdte
Die Wucht der derzeitigen Urbanisierungsdynamik und ihre Auswirkungen sind so groĂ, dass sich weltweit StĂ€dte, Stadtgesellschaften, Regierungen und Internationale Organisationen diesem Trend stellen mĂŒssen. Ein âWeiter so wie bisherâ, wĂŒrde ohne gestaltende Urbanisierungspolitik zu einer nicht-nachhaltigen Welt-StĂ€dte-Gesellschaft fĂŒhren. Nur wenn StĂ€dte und Stadtgesellschaften ausreichend handlungsfĂ€hig werden, können sie ihre Kraft fĂŒr eine nachhaltige Entwicklung entfalten: In den StĂ€dten wird sich entscheiden, ob die GroĂe Transformation zur Nachhaltigkeit gelingt. In diesem Buch werden die Erfolgsbedingungen dafĂŒr diskutiert
Humanity on the move: Unlocking the transformative power of cities
The momentum of urbanization and its impacts are so massive that we must face up to this trend. In view of the existing cognitive, technical, economic and institutional path dependencies, a policy of business as usual â i.e. an unstructured, quasi-automatic urbanization â would lead to a non-sustainable âworld cities societyâ. Only if cities and urban societies are sufficiently empowered can they make use of the opportunities for sustainability and successfully follow the urban transformation pathways. The success or failure of the Great Transformation will be decided in the cities. The WBGU discusses the relevant conditions for the success of this transformation in this report
Agua Salud alphavirus infection, dissemination and transmission in Aedes aegypti mosquitoes
Mosquitoes are competent vectors for many important arthropod-borne viruses (arboviruses). In addition to arboviruses, insect-specific viruses (ISV) have also been discovered in mosquitoes. ISVs are viruses that replicate in insect hosts but are unable to infect and replicate in vertebrates. They have been shown to interfere with arbovirus replication in some cases. Despite the increase in studies on ISVâarbovirus interactions, ISV interactions with their hosts and how they are maintained in nature are still not well understood. In the present study, we investigated the infection and dissemination of the Agua Salud alphavirus (ASALV) in the important mosquito vector Aedes aegypti through different infection routes (per oral infection, intrathoracic injection) and its transmission. We show here that ASALV infects the female Ae. aegypti and replicates when mosquitoes are infected intrathoracically or orally. ASALV disseminated to different tissues, including the midgut, salivary glands and ovaries. However, we observed a higher virus load in the brain than in the salivary glands and carcasses, suggesting a tropism towards brain tissues. Our results show that ASALV is transmitted horizontally during adult and larval stages, although we did not observe vertical transmission. Understanding ISV infection and dissemination dynamics in Ae. aegypti and their transmission routes could help the use of ISVs as an arbovirus control strategy in the future
Der virtuelle Raum als Double - oder: zur Persistenz hierarchischer Gesellschaftsstruktur im Netz
Der raumsoziologische Beitrag diskutiert, was die mit der Technologie des Internets einhergehende Virtualisierung fĂŒr eine sich Ă€ndernde Gesellschaftsordnung bedeuten kann. Anhand ihres methodologischen RaumZeit-Modells zeigt die Autorin, dass die virtuelle RealitĂ€t des Internets eine doppelte materiale Gestalt hervorbringt. Als Double ergĂ€nzt der virtuelle Raum den realen. Die Ausgangsannahme lautet, dass die bĂŒrgerlich-moderne Gesellschaft Raum als ZweidimensionalitĂ€t und als BehĂ€lter sowie Zeit als messbar und linear hervorgebracht hat. Entgegen vielfach geĂ€uĂerter EinschĂ€tzungen revolutioniert das Internet nicht Vorstellungen und Praxis von Raum und Zeit, sondern es perfektioniert die bĂŒrgerliche Konstruktion des ideal beherrschbaren Lebens. Der Hypothese folgend, dass die neuen virtuellen RealitĂ€ten verĂ€nderte MaterialitĂ€ten etablieren, werden verschiedene Zukunftsszenarien am Beispiel des GeschlechterverhĂ€ltnisses durchgespielt
Molecular Cloning, Characterization and Expression Analysis of Two Members of the Pht1 Family of Phosphate Transporters in Glycine max
BACKGROUND: Phosphorus is one of the macronutrients essential for plant growth and development. The acquisition and translocation of phosphate are pivotal processes of plant growth. In a large number of plants, phosphate uptake by roots and translocation within the plant are presumed to occur via a phosphate/proton cotransport mechanism. PRINCIPAL FINDINGS: We cloned two cDNAs from soybean (Glycine max), GmPT1 and GmPT2, which show homology to the phosphate/proton cotransporter PHO84 from the budding yeast Saccharomyces cerevisiae. The amino acid sequence of the products predicted from GmPT1 and GmPT2 share 61% and 63% identity, respectively, with the PHO84 in amino acid sequence. The deduced structure of the encoded proteins revealed 12 membrane-spanning domains with a central hydrophilic region. The molecular mass values are âŒ58.7 kDa for GmPT1 and âŒ58.6 kDa for GmPT2. Transiently expressed GFP-protein fusions provide direct evidence that the two Pi transporters are located in the plasma membrane. Uptake of radioactive orthophosphate by the yeast mutant MB192 showed that GmPT1 and GmPT2 are dependent on pH and uptake is reduced by the addition of uncouplers of oxidative phosphorylation. The K(m) for phosphate uptake by GmPT1 and GmPT2 is 6.65 mM and 6.63 mM, respectively. A quantitative real time RT-PCR assay indicated that these two genes are expressed in the roots and shoots of seedlings whether they are phosphate-deficient or not. Deficiency of phosphorus caused a slight change of the expression levels of GmPT1 and GmPT2. CONCLUSIONS: The results of our experiments show that the two phosphate transporters have low affinity and the corresponding genes are constitutively expressed. Thereby, the two phosphate transporters can perform translocation of phosphate within the plant
Functional characterization of the PHT1 family transporters of foxtail millet with development of a novel Agrobacterium-mediated transformation procedure
Phosphate is an essential nutrient for plant growth and is acquired from the environment and distributed within the plant in part through the action of phosphate transporters of the PHT1 family. Foxtail millet (Setaria italica) is an orphan crop essential to the food security of many small farmers in Asia and Africa and is a model system for other millets. A novel Agrobacterium-mediated transformation and direct plant regeneration procedure was developed from shoot apex explants and used to downregulate expression of 3 members of the PHT1 phosphate transporter family SiPHT1;2 SiPHT1;3 and SiPHT1;4. Transformants were recovered with close to 10% efficiency. The downregulation of individual transporters was confirmed by RT-PCR. Downregulation of individual transporters significantly reduced the total and inorganic P contents in shoot and root tissues and increased the number of lateral roots and root hairs showing they have non-redundant roles. Downregulation of SiPHT1;2 had the strongest effect on total and inorganic P in shoot and root tissues. Complementation experiments in S. cerevisiae provide evidence for the ability of SiPHT1;1, 1;2, 1;3, 1;7 and 1;8 to function as high affinity Pi transporters. This work will aid development of improved millet varieties for global food security
Development and justice through transformation: The Four Big âIâs. Special Report
2015 saw a historic double success for sustainability and climate policy. The 2030 Agenda for Sustainable Development, with its Sustainable Development Goals (SDGs), and the Paris Agreement on climate Âprotection establish a system of ambitious policy goals for the world. The group of twenty major Âindustrialized and emerging economies (G20) now needs to resolutely advance implementation of both agreements, seizing the opportunity of this âGreat Transformationâ to sustainability as a unique Âmodernization project that could offer substantial economic development opportunities. Complete Âdecarbonization of the world economy, which is necessary to avoid the gravest climate risks, can only be achieved by profoundly Âtransforming energy systems and other high-emissions infrastructures. This transformation could inspire ÂInnovation and channel Investment into sustainability and climate protection, and into the kinds of Âsustainable Infrastructures that need to be Âestablished and expanded. At the same time, the transformation could combat inequality and promote ÂInclusion within societies and globally, thus becoming an equity project
Pressure adaptation is linked to thermal adaptation in salt-saturated marine habitats
The present study provides a deeper view of protein
functionality as a function of temperature, salt and
pressure in deep-sea habitats. A set of eight different
enzymes from five distinct deep-sea (3040â4908 m
depth), moderately warm (14.0â16.5°C) biotopes,
characterized by a wide range of salinities (39â348
practical salinity units), were investigated for this
purpose. An enzyme from a âsuperficialâ marine
hydrothermal habitat (65°C) was isolated and characterized
for comparative purposes. We report here the
first experimental evidence suggesting that in saltsaturated
deep-sea habitats, the adaptation to high
pressure is linked to high thermal resistance (P
value = 0.0036). Salinity might therefore increase the
temperature window for enzyme activity, and possibly
microbial growth, in deep-sea habitats. As an
example, Lake Medee, the largest hypersaline deepsea
anoxic lake of the Eastern Mediterranean Sea,
where the water temperature is never higher than
16°C, was shown to contain halopiezophilic-like
enzymes that are most active at 70°C and with denaturing
temperatures of 71.4°C. The determination of
the crystal structures of five proteins revealed
unknown molecular mechanisms involved in protein
adaptation to poly-extremes as well as distinct active
site architectures and substrate preferences relative
to other structurally characterized enzymes.European Community project MAMBA (FP7-KBBE-2008-226977). This grant BIO2011-25012 from the Spanish Ministry of Economy and Competitiveness (formerly MICINN). European Commission for âMicroB3â grant (FP7-OCEAN.2011-2 (contract Nr
287589)). Government of Canada through Genome Canada
and the Ontario Genomics Institute (grant 2009-OGI-ABC-1405) and
U.S. National Institutes of Health (grants GM074942 and GM094585). Midwest Center for Structural Genomics).http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1462-2920hb2016Biochemistr
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