199 research outputs found

    Quantum computers based on electron spins controlled by ultra-fast, off-resonant, single optical pulses

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    We describe a fast quantum computer based on optically controlled electron spins in charged quantum dots that are coupled to microcavities. This scheme uses broad-band optical pulses to rotate electron spins and provide the clock signal to the system. Non-local two-qubit gates are performed by phase shifts induced by electron spins on laser pulses propagating along a shared waveguide. Numerical simulations of this scheme demonstrate high-fidelity single-qubit and two-qubit gates with operation times comparable to the inverse Zeeman frequency.Comment: 4 pages, 4 figures, introduction is clarified, the section on two-qubit gates was expanded and much more detail about gate fidelities is given, figures were modified, one figure replaced with a figure showing gate fidelities for relevant parameter

    A new fungal isolates applicated to bovine skin in beamhouse processes

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    Content: Beamhouse area is an important step in leather technology, either in the final quality of the leather or in the contribution to the contamination of effluents. In unhairing process, the use of enzymes as sulphide assistants can reduce sulfhidric gas emissions to reach permitted levels for health. The characterization of enzymatic extracts allows controlling the proteolysis mechanism so that its action does not attack the reticular structure. A new fungal enzyme extracts were applied on submerged bovine skin in the soaking and unhairing steps. Fungal strains were isolated from alkaline soils of coast of Buenos Aires province and enzymatic extracts (EE) were obtained by submerged culture with bovine hair waste from hair-saving unhairing process as inducer in the production of keratinolytic enzymes. This activity was determined by quantitative test and the most productive strains were selected: Clonostachys rosea (CR), Emericellopsis minima (EM), Paecilomyces lilacinus (PL), Penicillium sp (Psp), Fusarium oxysporum (FO), Acremonium sp (A) and an unidentified filamentous fungus strain with sterile yellow mycelium (SYM). EE were tested at laboratory scale on submerged bovine skin with biocide 0,2% w/w wet skin, anionic 0,1% and non-ionic 0,5% tensioactives in soaking and unhairing respectively and incubated 48 h at 37°C in shaking thermostatic bath. Controls were tested in the same conditions without EE. Morphological changes in the skin were observed by scanning electron microscopy (SEM). Pieces of bovine skin were fixed, post-fixed in formaldehyde 4% and dehydrated in alcohol solutions, treated by critical point drying, metalized and analyzed in SEM. It was observed with FO, EM and A extracts empty hair follicles, absence of epidermis, papillary layer of dermis exposed, hairs enclosed by detached hair follicle sheaths and collagen fibers without characteristic patterns. With PL and SYM EE some hairs were detached while Psp and CR EE did not produce changes. In control samples epidermis and hair without modification were observed. In the quantitative test, keratinolytic activities increased in the following order: A<PL<Psp<CR<FO<SYM. Submerged culture with keratin as inductor produced keratinolytic enzymes useful for unhairing step. Fusarium oxysporum extract showed the greatest effect on the skin, thus the relationship between keratinolytic activity and depilatory effect was found. However, it is necessary to find the optimal conditions to avoid the damage of collagen and enable its application as a sustainable technology. Take-Away: A new fungal isolates applicated to bovine skin in beamhouse processe

    Ultrahigh-throughput-directed enzyme evolution by absorbance-activated droplet sorting (AADS)

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    Ultrahigh-throughput screening, in which members of enzyme libraries compartmentalized in water-in-oil emulsion droplets are assayed, has emerged as a powerful format for directed evolution and functional metagenomics but is currently limited to fluorescence readouts. Here we describe a highly efficient microfluidic absorbance-activated droplet sorter (AADS) that extends the range of assays amenable to this approach. Using this module, microdroplets can be sorted based on absorbance readout at rates of up to 300 droplets per second (i.e., >1 million droplets per hour). To validate this device, we implemented a miniaturized coupled assay for NAD(+)-dependent amino acid dehydrogenases. The detection limit (10 μM in a coupled assay producing a formazan dye) enables accurate kinetic readouts sensitive enough to detect a minimum of 1,300 turnovers per enzyme molecule, expressed in a single cell, and released by lysis within a droplet. Sorting experiments showed that the AADS successfully enriched active variants up to 2,800-fold from an overwhelming majority of inactive ones at ∼100 Hz. To demonstrate the utility of this module for protein engineering, two rounds of directed evolution were performed to improve the activity of phenylalanine dehydrogenase toward its native substrate. Fourteen hits showed increased activity (improved >4.5-fold in lysate; kcat increased >2.7-fold), soluble protein expression levels (up 60%), and thermostability (Tm, 12 °C higher). The AADS module makes the most widely used optical detection format amenable to screens of unprecedented size, paving the way for the implementation of chromogenic assays in droplet microfluidics workflows.This research was funded by the Engineering and Physical Sciences Research Council (studentship to RH and an Impact Acceleration Account Partnership Development Award), the Biological and Biotechnological Research Council (BBSRC) and Johnson Matthey. SE and MF were supported by postdoctoral Marie-Curie fellowships

    Purification and characterization of a polygalacturonase produced by Wickerhamomyces anomalus

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    The aim of this work was to study the purification and physicochemical properties of an endo-polygalacturonase (PG) produced by Wickerhamomyces anomalus isolated from the citrus fruit peels. The enzyme was purified to homogeneity from the culture filtrate of W. anomalus grown on the yeast nitrogen base medium with glucose as carbon and energy source and citrus pectin as inductor. After anion-exchange chromatography and gel filtration chromatography, PG activity was eluted as a single peak, yielding 21% of the original activity. After dialysis and cation-exchange chromatography, only one fraction with PG activity was obtained, recovering 56% of initial enzyme activity and 1.3-fold increase in specific activity. The molecular weight of the enzyme was estimated as 43 kDa by the SDS-PAGE. The enzyme exhibited maximal activity at pH 4.2 and was stable over a pH range from 3.5 to 6.0 and up to 49°C for 10 h. The Vmax and Km values with polygalacturonic acid as substrate were 0.26 mmol/L. min and 0.173 mg/mL, respectively. Cations such as Cu+2, Fe+3, Mg+2, Mn+2 and Zn+2 did not show any significant effect on PG activity but K+ and Ca+2 reduced it. The purified PG was able to macerate cassava tissues.Centro de Investigación y Desarrollo en Fermentaciones Industriale

    Ultrahigh-throughput-directed enzyme evolution by absorbance-activated droplet sorting (AADS)

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    This is the final version. Available from National Academy of Sciences via the DOI in this recordUltrahigh-throughput screening, in which members of enzyme libraries compartmentalized in water-in-oil emulsion droplets are assayed, has emerged as a powerful format for directed evolution and functional metagenomics but is currently limited to fluorescence readouts. Here we describe a highly efficient microfluidic absorbance-activated droplet sorter (AADS) that extends the range of assays amenable to this approach. Using this module, microdroplets can be sorted based on absorbance readout at rates of up to 300 droplets per second (i.e., >1 million droplets per hour). To validate this device, we implemented a miniaturized coupled assay for NAD+-dependent amino acid dehydrogenases. The detection limit (10 μM in a coupled assay producing a formazan dye) enables accurate kinetic readouts sensitive enough to detect a minimum of 1,300 turnovers per enzyme molecule, expressed in a single cell, and released by lysis within a droplet. Sorting experiments showed that the AADS successfully enriched active variants up to 2,800-fold from an overwhelming majority of inactive ones at ∼100 Hz. To demonstrate the utility of this module for protein engineering, two rounds of directed evolution were performed to improve the activity of phenylalanine dehydro-genase toward its native substrate. Fourteen hits showed increased activity (improved >4.5-fold in lysate; kcat increased >2.7-fold), soluble protein expression levels (up 60%), and thermostability (Tm, 12°C higher). The AADS module makes the most widely used optical detection format amenable to screens of unprecedented size, paving the way for the implementation of chromogenic assays in droplet microfluidics workflows.Biotechnology and Biological Sciences Research CouncilEuropean Research CouncilEngineering and Physical Sciences Research CouncilEuropean Commissio

    5-Keto-D-fructose production from sugar alcohol by isolated wild strain Gluconobacter frateurii CHM 43

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    Gluconobacter frateurii: CHM 43 have D-mannitol dehydrogenase (quinoprotein glycerol dehydrogenase) and flavoprotein D-fructose dehydrogenase in the membranes. When the two enzymes are functional, D-mannitol is converted to 5-keto-D-fructose with 65% yield when cultivated on D-mannitol. 5-Keto-D-fructose production with almost 100% yield was realized with the resting cells. The method proposed here should give a smart strategy for 5-keto-D-fructose production.Fil: Adachi, Osao. Yamaguchi University; JapónFil: Nguyen, Thuy M.. Yamaguchi University; JapónFil: Hours, Roque Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaFil: Kataoka, Naoya. Yamaguchi University; JapónFil: Matsushita, Kazunobu. Yamaguchi University; JapónFil: Akakabe, Yoshihiko. Yamaguchi University; JapónFil: Yakushi, Toshiharu. Yamaguchi University; Japó

    Bladder cancer and occupational exposures

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    A hospital-based cas-referent study was carried out in Lyon with the purpose of generating hypotheses about the role of occupational exposures to 320 compounds in bladder carcinogenesis. Job histories were obtained by questionnaire for 116 cases and 232 reference patients with diseases other than cancer (one referent from the same hospital ward and one from another ward of the same hospital per case) ; the referents were matched for gender, hospital, age, and nationality. Systematic coding of exposures, with a blind analysis of job histories, was carried out by a team of experts in chemistry and occupational health. Significantly elevated odds ratios were observed for exposure to pyrolysis and combustion products [odds ratio (OR) 2.3, 95 % confidence interval (95 % CI) 1.0-4.0] when the general referents were used and for cutting fluids (OR 2.6, 95 % CI 1.2-5.4) when tobacco consumption was adjusted for. The latter was highest among the category consisting of blue-collar and unskilled workers, supervisors, and agricultural workers (OR 4.6, 95 % CI 2.0-10.6), while the odds ratio for the other category was 0.8 (95 % CI 0.3-2.7). An elevated odds ratio for exposure to inks was observed for the women (OR 14.0, 95 % CI 1.8-106.5) on the basis of 14 exposed cases, but confounding factors could have been responsible for this result. Odds ratios for several other exposures (rubber : OR 5.7, nitrates : OR 8.2, coke dust : OR 3.5, meat additives : OR 3.8) were also elevated, but not significantly so when based on a small number of exposed cases. The observations of this investigation should be tested in future studies, in particular since exposures to agents such as cutting fluids or pyrolysis products are ubiquitous in industrial settings and may present an important public health hazard. (Résumé d'auteur

    Production of polygalacturonase by Pichia anomala isolated in the province of Misiones

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    Las enzimas pécticas microbianas son de interés en la industria de alimentos principalmente por su uso en el procesamiento de frutas y vegetales (elaboración de vinos, extracción y clarificación de jugos de frutas, extracción de aceites vegetales, maceración de vegetales, etc.). Las pectinasas constituyen un complejo sistema enzimático que incluye: pectinesterasas (PE), poligalacturonasas (PG) y pectinliasas (PL). El objetivo del presente trabajo fue estudiar la cinética de producción de PG por Pichia anomala, levadura aislada en la provincia de Misiones, en medios de cultivos con diferentes concentraciones de la fuente de carbono y energía (FCE) y de la fuente de nitrógeno. Los cultivos se realizaron en Erlenmeyers de 500 ml conteniendo 95 ml de un medio compuesto por 5 ó 10 g/l de glucosa; 0, 5, 10 ó 20 g/l de pectina de citrus y 6,7 ó 13,4 g/l de Yeast Nitrogen Base (YNB). Los mismos se incubaron a 30 °C, con agitación (150 rpm). Se tomaron muestras cada 12 h hasta los 3 días. La biomasa se separó por centrifugación y el sobrenadante se mantuvo a -18 °C hasta su utilización para la medida de la actividad PG (por determinación de grupos reductores liberados a partir de ácido poligalacturónico, mediante el método DNS) y glucosa residual (mediante kit enzimático). En los medios conteniendo 5 g/l de glucosa como única FCE, la producción de PG fue despreciable. Al adicionar 5 ó 10 g/l de pectina, la enzima se expresó alcanzando valores de 60 UE/ml y 99 UE/ml, respectivamente, a las 30 h de cultivo, pero no se incrementó con 20 g/l de pectina. El incremento en la concentración de glucosa o de YNB resultó en un aumento de la actividad enzimática. Se obtuvieron valores de actividad PG de 168 UE/ml y 145 UE/ml en medios con 10 g/l de glucosa y 13,4 g/l de YNB, respectivamente. La mayor producción de biomasa se obtuvo entre las 24 y 30 h de cultivo en todas las experiencias, tiempo a partir del cual el microorganismo entró en fase estacionaria. Al aumentar la concentración de pectina no se observó un aumento en la cantidad de biomasa, esto indicaría que el microorganismo no es capaz de utilizar pectina como FCE pero si la necesita como inductor. La máxima producción de PG se obtuvo durante las primeras 24-30 h de cultivo (crecimiento exponencial). El pH disminuyó en el transcurso de las fermentaciones en todos los medios estudiados (probablemente los ácidos orgánicos producidos no fueron metabolizados por el microorganismo). La glucosa fue rápidamente metabolizada ya que luego de 24-30 h se obtuvieron valores despreciables en todos los medios ensayados, coincidiendo con el máximo crecimiento microbiano. Pichia anomala es capaz de producir PG en medios de cultivo con glucosa, pectina e YNB. La producción de PG se realizó durante la fase de crecimiento exponencial, no es reprimida por glucosa y es inducida por pectina.Microbial pectinases are very important in food industries mainly for their use in fruit and vegetable processing (winemaking, extraction and clarification of fruit juices, vegetable oil extraction, maceration of vegetables, etc.). Pectinases constitute a complex pool of enzymes that include: pectinesterases (PE), polygalacturonases (PG) and pectinlyases (PL). The objective of the present work was to study the kinetics of PG production by Pichia anomala, a wild yeast isolated in the province of Misiones, in culture media with different concentrations of carbon and energy source (CES) and nitrogen source. Cultures were grown in 500 ml-Erlenmeyer flasks with 95 ml of a medium containing 5 or 10 g/l glucose; 0, 5, 10 or 20 g/l citrus pectin and 6.7 or 13.4 g/l Yeast Nitrogen Base (YNB), at 30 °C, in a rotary shaker (150 rpm). Samples were taken every 12 h up to 3 days. Biomass was separated by centrifugation and the supernatant was kept at -18 °C until used for PG assay (by measuring the reducing groups released from polygalacturonic acid by DNS method) and residual glucose (by enzymatic kit). In culture media containing 5 g/l glucose as a sole CES, PG production was negligible. The enzyme was expressed by addition of 5 or 10 g/l of pectin, reaching values of 60 EU/ml and 99 EU/ml, respectively, at 30 h of cultivation, but enzyme activity did not increase with 20 g/l of pectin. The increase in the concentration of glucose or YNB resulted in an increase in enzyme activity. PG activity values of 168 EU/ml and 145 EU/ml were found in media with 10 g/l glucose and 13.4 g/l of YNB, respectively. The highest biomass production was obtained between 24 and 30 h of culture in all experiments, time at which the microorganism reached the stationary phase. Higher concentration of pectin did not result in an increase in biomass concentration; this would indicate that the microorganism is not capable of using pectin as CES but it is necessary as enzyme inducer. Maximum PG production was obtained during the first 24-30 h of culture (exponential growth). pH decreased during fermentation in all media studied (probably organic acids produced were not metabolized by the microorganism). Glucose was rapidly metabolized; negligible values were found in all media tested after 24-30 h of culture, coinciding with the maximum of microbial growth. Pichia anomala is capable of producing PG in culture media containing glucose, pectin and YNB. PG production takes place during the exponential growth phase; its production is not repressed by glucose and is induced by pectin.Centro de Investigación y Desarrollo en Criotecnología de Alimento
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