Influence of the Cu-Te composition and microstructure on the resistive switching of Cu-Te/Al(2)O(3)/Si cells

In this letter, we explore the influence of the Cu(x)Te(1-x) layer composition (0.2 0.7 leads to large reset power, similar to pure-Cu electrodes, x < 0.3 results in volatile forming properties. The intermediate range 0.5< x < 0.7 shows optimum memory properties, featuring improved control of filament programming using <5 mu A as well as state stability at 85 degrees C. The composition-dependent programming control and filament stability are closely associated with the phases in the Cu(x)Te(1-x) layer and are explained as related to the chemical affinity between Cu and Te. (C) 2011 American Institute of Physics. [doi: 10.1063/1.3621835

Simple and efficient expression of Agaricus meleagris pyranose dehydrogenase in Pichia pastoris

Pyranose dehydrogenase (PDH) is a fungal flavin-dependent sugar oxidoreductase that is highly interesting for applications in organic synthesis or electrochemistry. The low expression levels of the filamentous fungus Agaricus meleagris as well as the demand for engineered PDH make heterologous expression necessary. Recently, Aspergillus species were described to efficiently secrete recombinant PDH. Here, we evaluate recombinant protein production with expression hosts more suitable for genetic engineering. Expression in Escherichia coli resulted in no soluble or active PDH. Heterologous expression in the methylotrophic yeast Pichia pastoris was investigated using two different signal sequences as well as a codon-optimized sequence. A 96-well plate activity screening for transformants of all constructs was established and the best expressing clone was used for large-scale production in 50-L scale, which gave a volumetric yield of 223 mg L−1 PDH or 1,330 U L−1 d−1 in space–time yield. Purification yielded 13.4 g of pure enzyme representing 95.8% of the initial activity. The hyperglycosylated recombinant enzyme had a 20% lower specific activity than the native enzyme; however, the kinetic properties were essentially identical. This study demonstrates the successful expression of PDH in the eukaryotic host organism P. pastoris paving the way for protein engineering. Additionally, the feasibility of large-scale production of the enzyme with this expression system together with a simplified purification scheme for easy high-yield purification is shown

Discovery and characterization of a new family of lytic polysaccharide monooxygenases

Lytic polysaccharide monooxygenases (LPMOs) are a recently discovered class of enzymes capable of oxidizing recalcitrant polysaccharides. They are attracting considerable attention owing to their potential use in biomass conversion, notably in the production of biofuels. Previous studies have identified two discrete sequence-based families of these enzymes termed AA9 (formerly GH61) and AA10 (formerly CBM33). Here, we report the discovery of a third family of LPMOs. Using a chitin-degrading exemplar from Aspergillus oryzae, we show that the three-dimensional structure of the enzyme shares some features of the previous two classes of LPMOs, including a copper active center featuring the 'histidine brace' active site, but is distinct in terms of its active site details and its EPR spectroscopy. The newly characterized AA11 family expands the LPMO clan, potentially broadening both the range of potential substrates and the types of reactive copper-oxygen species formed at the active site of LPMOs

Characterization of an Alkali- and Halide-Resistant Laccase Expressed in E. coli: CotA from <i>Bacillus clausii</i>

The limitations of fungal laccases at higher pH and salt concentrations have intensified the search for new extremophilic bacterial laccases. We report the cloning, expression, and characterization of the bacterial cotA from Bacillus clausii, a supposed alkalophilic ortholog of cotA from B. subtilis. Both laccases were expressed in E. coli strain BL21(DE3) and characterized fully in parallel for strict benchmarking. We report activity on ABTS, SGZ, DMP, caffeic acid, promazine, phenyl hydrazine, tannic acid, and bilirubin at variable pH. Whereas ABTS, promazine, and phenyl hydrazine activities vs. pH were similar, the activity of B. clausii cotA was shifted upwards by ~0.5-2 pH units for the simple phenolic substrates DMP, SGZ, and caffeic acid. This shift is not due to substrate affinity (K(M)) but to pH dependence of catalytic turnover: The k(cat) of B. clausii cotA was 1 s⁻¹ at pH 6 and 5 s⁻¹ at pH 8 in contrast to 6 s⁻¹ at pH 6 and 2 s⁻¹ at pH 8 for of B. subtilis cotA. Overall, k(cat)/K(M) was 10-fold higher for B. subtilis cotA at pH(opt). While both proteins were heat activated, activation increased with pH and was larger in cotA from B. clausii. NaCl inhibited activity at acidic pH, but not up to 500-700 mM NaCl in alkaline pH, a further advantage of the alkali regime in laccase applications. The B. clausii cotA had ~20 minutes half-life at 80°C, less than the ~50 minutes at 80°C for cotA from B. subtilis. While cotA from B. subtilis had optimal stability at pH~8, the cotA from B. clausii displayed higher combined salt- and alkali-resistance. This resistance is possibly caused by two substitutions (S427Q and V110E) that could repel anions to reduce anion-copper interactions at the expense of catalytic proficiency, a trade-off of potential relevance to laccase optimization

Study of the sensitisation process of a duplex stainless steel (UNS 1.4462) by means of confocal microscopy and localised electrochemical techniques

When duplex stainless steels are heated, they can become sensitised and intermetallic phases can appear. In this work, samples of duplex stainless steel, UNS 1.4462, have been heated (850 degrees C for 1 and 2 h) in a heating unit that can be accommodated to a confocal microscope in order to study the morphological changes in-situ. The electrochemical behaviour of the samples has been analysed by means of conventional and localised electrochemical techniques. According to the results, there is a general decrease in the steel resistance to localised corrosion; this decrease can be related to defects in the formed passive film.We wish to express our gratitude to MICINN (CTQ2009-07518), to Universitat Politecnica de Valencia (CEI-01-11), to the Generalitat Valenciana for its help in the CLSM acquisition (MY08/ISIRM/S/100), to Professor Alda Simoes and Andreia Marques for the help on LEIS measurements and to Dr. Asuncion Jaime for her translation assistance.Leiva García, R.; Fernandes, JS.; Muñoz-Portero, M.; Garcia-Anton, J. (2015). Study of the sensitisation process of a duplex stainless steel (UNS 1.4462) by means of confocal microscopy and localised electrochemical techniques. Corrosion Science. 94:327-341. https://doi.org/10.1016/j.corsci.2015.02.016S3273419