Disrupting the methionine biosynthetic pathway in Candida guilliermondii: characterization of the MET2 gene as counter‐selectable marker

Candida guilliermondii (teleomorph Meyerozyma guilliermondii) is an ascomycetous species belonging to the fungal CTG clade. This yeast remains actively studied as a result of its moderate clinical importance and most of all for its potential uses in biotechnology. The aim of the present study was to establish a convenient transformation system for C. guilliermondii by developing both a methionine auxotroph recipient strain and a functional MET gene as selection marker. We first disrupted the MET2 and MET15 genes encoding homoserine‐O‐acetyltransferase and O‐acetylserine O‐acetylhomoserine sulphydrylase, respectively. The met2 mutant was shown to be a methionine auxotroph in contrast to met15 which was not. Interestingly, met2 and met15 mutants formed brown colonies when cultured on lead‐containing medium, contrary to the wild‐type strain, which develop as white colonies on this medium. The MET2 wild‐type allele was successfully used to transfer a yellow fluorescent protein (YFP) gene‐expressing vector into the met2 recipient strain. In addition, we showed that the loss of the MET2‐containing YFP‐expressing plasmid can be easily observed on lead‐containing medium. The MET2 wild‐type allele, flanked by two short repeated sequences, was then used to disrupt the LYS2 gene (encoding the α‐aminoadipate reductase) in the C. guilliermondii met2 recipient strain. The resulting lys2 mutants displayed, as expected, auxotrophy for lysine. Unfortunately, all our attempts to pop‐out the MET2 marker (following the recombination of the bordering repeat sequences) from a target lys2 locus were unsuccessful using white/brown colony colour screening. Nevertheless, this MET2 transformation/disruption system represents a new versatile genetic tool for C. guilliermondii. Copyright © 2014 John Wiley & Sons, Ltd.     

Understanding of Void Formation in Cu/Sn-Sn/Cu System During Transient Liquid Phase Bonding Process Through Diffusion Modeling

Metallurgical and Materials Transactions B - Transient Liquid Phase (TPL) bounding of Sn foil sandwiched between two Cu foils involves, in the temperature range above the melting point of Sn...     

Optimization of the formation of vinyldithiins, therapeutic compounds from garlic

Allyl sulfides, ajoenes and vinyldithiins are the three main groups of volatile organosulfur compounds that are formed when garlic is crushed. The manner garlic is processed (nature of the extraction medium, temperature…) has a major influence on their relative proportion and the amounts produced. It has been proven recently that the vinyldithiins are at the origin of garlic’s capacity to prevent adipocytes development. Their incorporation in garlic-based nutraceuticals is thus particularly interesting. In this context, this work aims to optimize the production of vinyldithiins from garlic. After having determined the best garlic origin (Spanish) and the best oil for the extraction (olive or sunflower oil), the extraction conditions were optimized (1/2 (w/w garlic oil), 37 °C, 6 h) and 133 mg of vinyldithiins was obtained from 100 g of fresh garlic. Carrying out the extraction under microwave irradiation allowed increasing the yield 3.6-fold (yield 486 mg of vinyldithiins from 100 g of fresh garlic). This study may also contribute to the development of new garlic derived high value products by enhancing the comprehension of their formation.     

Microstructure and texture of copper/niobium composites processed by ECAE

The structure refinement is a challenge for conductors used for the winding of resistive coils producing non-destructive pulsed magnetic fields over 80T. These nanocomposite conductors composed of a conducting multiscale Cu matrix embedding Nb reinforcing nanofilaments are usually manufactured by using a Severe Plastic Deformation (SPD) process based on hot extrusion, accumulative cold drawing and bundling (ADB) steps [Thilly et al. Philos Mag A 82:925, 2002]. Equal-channel angular extrusion (ECAE) is investigated here as an alternative route since it may provide faster refinement to obtain the ultrafine structure needed for optimized electrical and mechanical properties of the conductors. Therefore, copper-niobium specimens obtained by hot extrusion were processed by ECAE at room temperature. The specific die of the ECAE tool used here is constituted by a round channel with three angles corresponding to a total equivalent strain of about 2.5. Deformed samples were examined by optical microscopy and characterized by hardness profiles and x-ray diffraction (texture pole figures). After one ECAE-pass, the shape of the samples is modified but no trace of damage appeared at the Cu-Nb interfaces. An increase of the hardness values localized in the copper matrix is revealed whereas the hardness of the niobium remains unchanged. Prior to ECAE, the hot extrusion process induced a fibre texture for both copper and niobium. Two fibre texture components were observed for copper: <111> and <200>. A single <110> fibre texture component is evidenced for the niobium. After ECAE a significant variation of the texture is observed in relation with the strong shear induced by this process.     

Circularly polarized luminescence microscopy for the imaging of charge and spin diffusion in semiconductors

Room temperature electronic diffusion is studied in 3?μm thick epitaxial p(+) GaAs lift-off films using a novel circularly polarized photoluminescence microscope. The method is equivalent to using a standard optical microscope and provides a contactless means to measure both the charge (L) and spin (L(s)) diffusion lengths simultaneously. The measured values of L and L(s) are in excellent agreement with the spatially averaged polarization and a sharp reduction in these two quantities (L from 21.3 to 1.2?μm and L(s) from 1.3 to 0.8?μm) is found with increasing surface recombination velocity. Outward diffusion results in a factor of 10 increase in the polarization at the excitation spot. The range of materials to which the technique can be applied, as well as a comparison with other existing methods for the measurement of spin diffusion, is discussed.     

Polycyclic aromatic hydrocarbons in soil and surface marine sediment near Jubany Station (Antarctica). Role of permafrost as a low-permeability barrier

Although Antarctica is still considered as one of the most pristine areas of the world, the growing tourist and fisheries activities as well as scientific operations and their related logistic support are responsible for an increasing level of pollutants in this fragile environment. Soils and coastal sediments are significantly affected near scientific stations particularly by polycyclic aromatic hydrocarbons (PAHs). In this work sediment and soil were sampled in two consecutive summer Antarctic expeditions at Potter Cove and peninsula, in the vicinity of Jubany Station (South Shetland Islands). Two- and 3-ring PAHs (methylnaphthalene, fluorene, phenanthrene and anthracene) were the main compounds found in most sites, although total PAH concentrations showed relatively low levels compared with other human-impacted areas in Antarctica. Pattern distribution of PAHs observed in samples suggested that low-temperature combustion processes such as diesel motor combustion and open-field garbage burning are the main sources of these compounds. An increase in PAH concentrations was observed from surface to depth into the active soil layer except for a unique sampling site where a fuel spill had been recently reported and where an inverted PAH concentration gradient was observed. The highest level was detected in the upper layer of permafrost followed by a sharp decrease in depth, showing this layer is acting as a barrier for downward PAH migration. When PAH levels in soil from both sampling programs were compared a significant decrease (p<0.01) was observed in summer 2005 (range at 75-cm depth: 12+/-1-153+/-22 ng/g) compared to summer 2004 (range at 75-cm depth: 162+/-15-1182+/-113 ng/g) whereas concentrations in surface sediment collected nearby the station PAHs increased drastically in 2005 (range: 36+/-3-1908+/-114 ng/g) compared to 2004 (range: 28+/-3-312+/-24 ng/g). Precipitation regime and water run off suggest that an important wash out of soil-PAHs occurred during the interval time between samplings. Results showed that the present PAH contamination level of Jubany Station is relatively low compared to other reported cases in Antarctica but also suggests that an increase in rain and in thawing processes caused by the global warming could result in an important soil-associated PAH mobilization with unpredictable consequences for the biota of Potter Cove.     

Simple Fabrication and Characterization of Discontinuous Carbon Fiber Reinforced Aluminum Matrix Composite for Lightweight Heat Sink Applications

The constant increase in power and heat flux densities encountered in electronic devices fuels a rising demand for lightweight heat sink materials with suitable thermal properties.In this study,discontinuous pitch-based carbon fiber reinforced aluminum matrix(Al-CF) composites with aluminum–silicon alloy(Al–Si) were fabricated through hot pressing.The small amount of Al–Si contributed to enhance the sintering process in order to achieve fully dense Al–CF composites.A thermal conductivity and CTE of 258 W/(m K) and 7.0 9 10-6/K in the in-plane direction of the carbon fibers were obtained for a(Al95 vol%+ Al–Si5 vol%)-CF50 vol%composite.Carbon fiber provides the reducing of CTE while the conservation of thermal conductivity and weight of Al.The achieved CTEs satisfy the standard requirements for a heat sink material,which furthermore possess a specific thermal conductivity of 109 W cm3/(m K g).This simple process allows the low-cost fabrication of Al–CF composite,which is applicable for a lightweight heat sink material.     

Techno-Economic Analysis for Production of L-Arabitol from L-Arabinose

The process design for synthesis of arabitol by hydrogenation of arabinose on a supported ruthenium catalyst is described. Aspen HYSYS software was used to design an arabitol production plant for subsequent generation of hydrogen through aqueous phase reforming (APR). The process design included hydrogen recycling requiring a flash drum and recompression. The total costs of arabitol were estimated to be substantially dependent on the feedstock costs and are closely related to arabinose feed cost. Feasible production of hydrogen using APR requires efficient extraction of hemicellulose from lignocellulosic biomass and subsequent hydrolysis.