159 research outputs found
A Novel High-Cell-Density Protein Expression System Based on Ralstonia eutropha
We describe the development of a novel protein expression system based on the industrial fermentation organism Ralstonia eutropha (formerly known as Alcaligenes eutrophus) NCIMB 40124. This new system over- comes some of the shortcomings of traditional Escherichia coli-based protein expression systems, particularly the propensity of such systems to form inclusion bodies during high-level expression. Using a proteomics approach, we identified promoters that can be induced by simple process parameters or medium compositions in high-density cell culture or shake flasks, respectively. By combining newly developed molecular biological tools with a high-cell-density fermentation process, we were able to produce high levels (\u3e1 g/liter) of soluble, active organophosphohydrolase, a model enzyme prone to inclusion body formation in E. coli
Agro-materials : a bibliographic review
Facing the problems of plastic recycling and fossil resources exhaustion, the use of biomass to conceive new materials appears like a reasonable solution. Two axes of research are nowadays developed : on the one hand the synthesis of biodegradable plastics, whichever the methods may be, on the other hand the utilization of raw biopolymers, which is the object of this paper. From this perspective, the “plastic” properties of natural polymers, the caracteristics of the different classes of polymers, the use of charge in vegetable matrix and the possible means of improving the durability of these agro-materials are reviewed
Electrochemical growth of Co nanowires in ultra-high aspect ratio InP membranes: FFT-impedance spectroscopy of the growth process and magnetic properties
Modern insulation materials for warming of walls
Biodiversity hotspots understandably attract considerable conservation attention. However, deserts are rarely viewed as conservation priority areas, due to their relatively low productivity, yet these systems are home to unique species, adapted to harsh and highly variable environments. While global attention has been focused on hotspots, the world's largest tropical desert, the Sahara, has suffered a catastrophic decline in megafauna. Of 14 large vertebrates that have historically occurred in the region, four are now extinct in the wild, including the iconic scimitar-horned oryx (Oryx dammah). The majority has disappeared from more than 90% of their Saharan range, including addax (Addax nasomaculatus), dama gazelle (Nanger dama) and Saharan cheetah (Acinonyx jubatus hecki) - all now on the brink of extinction. Greater conservation support and scientific attention for the region might have helped to avert these catastrophic declines. The Sahara serves as an example of a wider historical neglect of deserts and the human communities who depend on them. The scientific community can make an important contribution to conservation in deserts by establishing baseline information on biodiversity and developing new approaches to sustainable management of desert species and ecosystems. Such approaches must accommodate mobility of both people and wildlife so that they can use resources most efficiently in the face of low and unpredictable rainfall. This is needed to enable governments to deliver on their commitments to halt further degradation of deserts and to improve their status for both biodiversity conservation and human well-being. Only by so-doing will deserts be able to support resilient ecosystems and communities that are best able to adapt to climate change. © 2013 John Wiley & Sons Ltd
Class III Polyhydroxybutyrate Synthase: Involvement in Chain Termination and Reinitiation
Trapping of Intermediates with Substrate Analog HBOCaA in the Polymerizations Catalyzer by Class III Polyhydroxybutyrate (PHB) Synthase from Allochromatium Vinosum
Polyhydroxybutyrate (PHB) synthases (PhaCs) catalyze the formation of biodegradable PHB polymers that are considered as an ideal alternative to petroleum-based plastics. To provide strong evidence for the preferred mechanistic model involving covalent and noncovalent intermediates, a substrate analog HBOCoA was synthesized chemoenzymatically. Substitution of sulfur in the native substrate HBCoA with an oxygen in HBOCoA enabled detection of (HB)nOCoA (n = 2–6) intermediates when the polymerization was catalyzed by wild-type (wt-)PhaECAv at 5.84 hr−1. This extremely slow rate is due to thermodynamically unfavorable steps that involve formation of enzyme-bound PHB species (thioesters) from corresponding CoA oxoesters. Synthesized standards (HB)nOCoA (n = 2–3) were found to undergo both reacylation and hydrolysis catalyzed by the synthase. Distribution of the hydrolysis products highlights the importance of the penultimate ester group as previously suggested. Importantly, the reaction between primed synthase [3H]-sT-PhaECAv and HBOCoA yielded [3H]-sTet-O-CoA at a rate constant faster than 17.4 s−1, which represents the first example that a substrate analog undergoes PHB chain elongation at a rate close to that of the native substrate (65.0 s−1). Therefore, for the first time with a wt-synthase, strong evidence was obtained to support our favored PHB chain elongation model
Properties, production, and applications of camelid single-domain antibody fragments
Camelids produce functional antibodies devoid of light chains of which the single N-terminal domain is fully capable of antigen binding. These single-domain antibody fragments (VHHs or Nanobodies®) have several advantages for biotechnological applications. They are well expressed in microorganisms and have a high stability and solubility. Furthermore, they are well suited for construction of larger molecules and selection systems such as phage, yeast, or ribosome display. This minireview offers an overview of (1) their properties as compared to conventional antibodies, (2) their production in microorganisms, with a focus on yeasts, and (3) their therapeutic applications
Effects of plurilingual teaching on grammatical development in early foreign-language learning
Role for cFMS in maintaining alternative macrophage polarization in SIV infection: implications for HIV neuropathogenesis
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