Sample records for azotobacter chroococcum purification

  1. Cultivation of the bacterium Azotobacter chroococcum for ...

    African Journals Online (AJOL)

    ... increase of bioprocess efficiency parameters (yield coefficient and productivity) were observed compared with the batch cultivation. On the basis of the obtained results, repeated batch technique appeared to be the most suitable for the bacterial biomass production at industrial scale. Key words: Azotobacter chroococcum, ...

  2. Nitrogen fixation and nitrogenase activity of Azotobacter chroococcum

    NARCIS (Netherlands)

    Brotonegoro, S.


    The purpose of the present investigation was to study the effect of some chemical, physical and biological factors on growth, efficiency of nitrogen fixation and nitrogenase activity of Azotobacter chroococcum.

    From biochemical studies with cell-free

  3. Cultivation of the bacterium Azotobacter chroococcum for ...

    African Journals Online (AJOL)



    Apr 18, 2011 ... affected by temperature that has to be in the range of 28 to 32°C .... The partial pressure of oxygen (pO2) was maintained at approximately 30% of ..... REFERENCES. Bakulin MK, Grudtsyna AS, Pletneva AY (2007). Biological fixation of nitrogen and growth of bacteria of the genus Azotobacter in liquid.

  4. Investigation on Strain Development of Azotobacter chroococcum through Chemical Mutagenesis for Indole Acetic Acid Production

    International Nuclear Information System (INIS)

    Ye Lin Win; Myo Myint Han; Myo Myint


    Two wild strains of Azotobacter chroococcum were mutagenized by various concentration of NTG (.005% , .0075% and .01%) for 1 hour. The IAA producing activity of 12 effective mutagenized strains were quantitatively measured by UV spectroscopic method. Mutagenized strains M1 and M2 from W1, and M8 and M9 from W2 were selected. Their IAA productivities were 300.498, 216.290, 238.436 and 190.856 ppm, respectively. M1and M9 were higher IAA productivity with greater quality. In germination, M1, M8 and M9 can promote the plant growth compare with commercial IAA.

  5. Melanin from the nitrogen-fixing bacterium Azotobacter chroococcum: a spectroscopic characterization.

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    Aulie Banerjee

    Full Text Available Melanins, the ubiquitous hetero-polymer pigments found widely dispersed among various life forms, are usually dark brown/black in colour. Although melanins have variety of biological functions, including protection against ultraviolet radiation of sunlight and are used in medicine, cosmetics, extraction of melanin from the animal and plant kingdoms is not an easy task. Using complementary physicochemical techniques (i.e. MALDI-TOF, FTIR absorption and cross-polarization magic angle spinning solid-state (13C NMR, we report here the characterization of melanins extracted from the nitrogen-fixing non-virulent bacterium Azotobacter chroococcum, a safe viable source. Moreover, considering dihydroxyindole moiety as the main constituent, an effort is made to propose the putative molecular structure of the melanin hetero-polymer extracted from the bacterium. Characterization of the melanin obtained from Azotobacter chroococcum would provide an inspiration in extending research activities on these hetero-polymers and their use as protective agent against UV radiation.

  6. Study the concentration of macroelements in forage mays (Zea mays L. (SC 704 as effected by inoculation with mycorrhizal fungi and Azotobacter chroococcum under different levels of nitrogen

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    M. Amirabadi


    Full Text Available Nitrogen and phosphorus are two necessary macronutrients for plant growth and yield. These two elements now will be added to soil by chemical fertilizers. This research has been carried out based on randomized completely block design with three replications at Markazi Provience Agricultural Research Station, Iran, during growing season of 2004-2005 to evaluate the effects of Azotobacter chroococcum and Mycorrhiza (Glomus intraradices as biofertilizers and urea as chemical fertilizers on concentrations of N, P, K, Na, Ca and crude protein (% in corn (Zea mays L. shoot tissues and dry matter of corn. Azotobacter chroococcum used as two levels (inoculated and uninoculated, mycorrhiza (Glomus intraradices in two levels (inoculated and uninoculated and urea in four levels (0, 75, 150 and 300 kg.ha-1. Results showed that Azotobacter chroococcum affected significantly all studied criteria except of K shoot concentration, but mycorrhizan (Glomus intraradices only had a increasing significantly effect on N, K, Na and Crude protein. The interaction between Azotobacter chroococcum and Mycorrhiza (Glomus intraradices had the most increasing effect on dry matter, N, Na and Crude protein. Therefore, based on our results it can be concluded that in order to prevent polluting the agricultural soil, environmental and other water supplies from nitrogen chemical fertilizers, application of Azotobacter chroococcum or combined with mycorrhizal fungi with 150 kg.ha-1 Urea is recommended.

  7. The effect of inoculation with Azotobacter chroococcum on microorganisms in rhizosphere and sugar beet yield in organic farming

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    Mrkovački Nastasija B.


    Full Text Available The effect on sugar beet yield parameters and microbiological soil status was studied using two techniques of sugar beet inoculation with strains of Azotobacter chroococcum. Cultivar “Drena” was used in the study, and field trial was set under the conditions of organic farming system in Bački Petrovac. A mixture of three strains of Azotobacter chroococcum was used as microbial fertilizer. Inoculation was performed by: (A incorporation of strains into soil before sowing; and (B repeated incorporation of strains into soil two weeks after sowing. PGP characterization of the strains confirmed the ability of producing indole-3-acetic acid (IAA from 12.63 μg ml-1 to 14.95 μg ml-1, nitrogen fixation, and P-solubilization. Positive effects on the number of azotobacter and free nitrogen fixers in rhizosphere were obtained by inoculation, as well as positive effects on the tested sugar beet yield parameters. The largest increase in root yield, yield of crystal sugar, and yield of polarised sugar compared with the control was obtained by repeated soil inoculation, ranging from 22 to 23%.


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    Soma Pal Saha


    Full Text Available Azotobacter chroococcum MAL-201 (MTCC 3853, a free-living nitrogen-fixing bacterium accumulated intracellular poly(3-hydroxybutyric acid [P(3HB] accounting 69% of cell dry weight (CDW when grown in nitrogrn-free Stockdale medium containing 2% (w/v glucose. It also produced copolymer of poly(3-hydroxybutyrate co-3-hydroxyvalerate [P(3HB-co-3HV] using glucose as primary carbon source and valerate cas cosubstrate. To make the polymer production cost effective four types of waste material of different origin were tested for growth and polymer production. Stockdale medium supplemented with 1% (w/v waste materials failed to yield good growth and polymer accumulation. Two–step cultivation was adopted for better growth and enhanced polymer accumulation. The candy factory waste was most suitable for synthesis of P(3HB accounting 17.8 and 40.58% using single and two-step cultivation conditions respectively. Wastes of domestic and poultry origin produced P(3HB-co-3HV with 3HV content 28.8 and 21.5 mol% respectively in two-step cultivation. Increase concentration of these wastes resulted in further upliftment of 3HV content of polymer with reduced growth and polymer accumulation. However, at optimum incubation the strain MAL-201 cells accumulated P(3HB 48.5% of CDW (at 40h from candy factory waste and P(3HB-co-3HV 24.75 % of CDW with 3HV 34.65 mol % from domestic waste. Intrinsic viscosity, molecular weight and thermal degradation of the polymers accumulated in the cells grown in glucose, glucose with valerate and glucose with waste were compared.

  9. Integrated approach for disease management and growth enhancement of Sesamum indicum L. utilizing Azotobacter chroococcum TRA2 and chemical fertilizer. (United States)

    Maheshwari, D K; Dubey, R C; Aeron, Abhinav; Kumar, Bhavesh; Kumar, Sandeep; Tewari, Sakshi; Arora, Naveen Kumar


    Azotobacter chroococcum TRA2, an isolate of wheat rhizosphere displayed plant growth promoting attributes including indole acetic acid, HCN, siderophore production, solubilization of inorganic phosphate and fixation of atmospheric nitrogen. In addition, it showed strong antagonistic effect against Macrophomina phaseolina and Fusarium oxysporum. It also caused degradation and digestion of cell wall components, resulting in hyphal perforations, empty cell (halo) formation, shrinking and lysis of fungal mycelia along with significant degeneration of conidia. Fertilizer adaptive variant strain of A. chroococcum TRA2 was studied with Tn5 induced streptomycin resistant transconjugants of wild type tetracycline-resistant TRA2 (designated TRA2(tetra+strep+)) after different durations. The strain was significantly competent in rhizosphere, as its population increased by 15.29 % in rhizosphere of Sesamum indicum. Seed bacterization with the strain TRA2 resulted in significant increase in vegetative growth parameters and yield of sesame over the non-bacterized seeds. However, application of TRA2 with half dose of fertilizers showed sesame yield almost similar to that obtained by full dose treatment. Moreover, the oil yield increased by 24.20 %, while protein yield increased by 35.92 % in treatment receiving half dose of fertilizer along with TRA2 bacterized seeds, as compared to untreated control.

  10. Co-inoculation with Mesorhizobium ciceri and Azotobacter chroococcum for improving growth, nodulation and yield of chickpea (Cicer arietinum L.

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    M.A. Qureshi


    Full Text Available Rhizobia have the exceptional ability to form nodules on roots or stems of leguminous plants. Free living diazotrophs promote the rhizobial efficiency by altering root architecture providing more niches for nodulation and thus enhance the N2-fixing ability of legumes. Field experiment was conducted to assess the co-inoculation potential of symbiotic i.e. Mesorhizobium ciceri and non-symbiotic diazotrophs i.e. Azotobacter chroococcum on the yield of chickpea. Chickpea seeds (cv. Bittle-98 were inoculated with peat-based inocula and sown following randomized complete block design with three replications. Two levels of nitrogen i.e. 30 (recommended and 15 kg ha-1 were applied as urea while P was applied at 60 kg ha-1 to all the treatments as single super phosphate. Results revealed that introduction of A. chroococcum had positive impact on chickpea with and without rhizobial inoculation and the effect was more prominent when applied in combination as compared to non-inoculated control at low nitrogen level. It was observed that inoculation with M. ciceri or A. chroococcum produced significant increase in biomass and grain yield but the response was more pronounced with co-inoculation i.e. 3456 and 1772 kg ha-1, respectively, as compared to control (2903 and 1489 kg ha-1, respectively at 15 kg N ha-1. Higher nodule number plant-1 and nodular mass was observed with co-inoculation (42 and 0.252 g plant-1. Percent N and P content in chickpea plant were higher in the co-inoculated treatments (1.683 and 0.283% than that of their respective controls. Similar trend was observed in grains except the rhizobial inoculation alone which produced higher N content (3.62% than coinoculation (3.59%. Percent N and available P in soil were also higher in the inoculated treatments. The results imply that co-inoculation with Mesorhizobium and Azotobacter could be a useful approach for improving growth, nodulation and yield of chickpea by reducing dependence on chemical


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    Full Text Available En la presente investigación, se analizó la influencia in vitro de los plaguicidas peletizados simultáneamente con el fungicida (principios activos: carboxin: 5,6-dihydro-2-methyl-N-phenyl-1,4-oxathiin-3 carboxamide y tiram: tetramethylthioperoxydicarbonic diamide y un insecticida (principio activo: malatión: S-1,2-Bis(ethoxycarbonylethyl-0,0 imethylthiophosphate en las semillas de algodón sobre la viabilidad del biofertilizante Monibac®-Corpoica con base en Azotobacter chroococcum AC1, cuando fueron aplicados conjuntamente en bandejas experimentales. Se analizó la práctica de recubrir las semillas con los agroquímicos como posible causa de descenso poblacional de la bacteria bajo este modelo de aplicación. Los resultados obtenidos por el Test de Duncan (pNo presente trabalho foi analisado a influencia in vitro dos plaguicidas peletizados simultáneamente com os fungicidas carboxin (5,6-dihydro-2-methyl-N-phenyl-1,4-oxathiin-Z carboxamide e tiram (tetramethylthioperoxydicarbonic diamide alem do inseticida malation (S-1,2 Bis(ethoxycarbonylethyl-0,0 imethylthiophosphate nas sementes de algodÃo sobre a viabilidade do biofertilizante Monibac - Corpoica contendo Azotobacter chroococcum, estirpe AC1, quando foram aplicados conjuntamente em bandejas experimentais. Foi analisado a pratica de recobrimento das sementes com os agroquímicos como possível causa da diminuição populacional das bacterias submetidas a este modelo de aplicação. Os resultados obtidos pelo teste Duncan (pIn this study, we analyzed the influence of pesticides in vitro pellet simultaneously with the fungicide (active ingredient: carboxin: 5,6-dihydro-2-methyl-N-phenyl-1,4-oxathiin-Z carboxamide and tiram: tetramethylthioperoxydicarbonic diamide and an insecticide (active ingredient: Malathion: S-1,2-Bis (ethoxycarbonyl ethyl-0, O-dimethylthiophosphate in cotton seeds on the viability of biofertiliser Monibac® -based Corpoica Azotobacter chroococcum ACT, when

  12. Azotobacter chroococcum as a potentially useful bacterial biofertilizer for cotton (Gossypium hirsutum): Effect in reducing N fertilization. (United States)

    Romero-Perdomo, Felipe; Abril, Jorge; Camelo, Mauricio; Moreno-Galván, Andrés; Pastrana, Iván; Rojas-Tapias, Daniel; Bonilla, Ruth

    The aim of this research was to evaluate whether the application of two plant growth-promoting (rhizo)bacteria might reduce nitrogen fertilization doses in cotton. We used strains Azotobacter chroococcum AC1 and AC10 for their proven ability to promote seed germination and cotton growth. These microorganisms were characterized by their plant growth-promoting activities. Then, we conducted a glasshouse study to evaluate the plant growth promoting ability of these strains with reduced doses of urea fertilization in cotton. Results revealed that both strains are capable of fixing nitrogen, solubilizing phosphorus, synthesizing indole compounds and producing hydrolytic enzymes. After 12 weeks, the glasshouse experiment showed that cotton growth was positively influenced due to bacterial inoculation with respect to chemical fertilization. Notably, we observed that microbial inoculation further influenced plant biomass (p<0.05) than nitrogen content. Co-inoculation, interestingly, exhibited a greater beneficial effect on plant growth parameters compared to single inoculation. Moreover, similar results without significant statistical differences were observed among bacterial co-inoculation plus 50% urea and 100% fertilization. These findings suggest that co-inoculation of A. chroococcum strains allow to reduce nitrogen fertilization doses up to 50% on cotton growth. Our results showed that inoculation with AC1 and AC10 represents a viable alternative to improve cotton growth while decreasing the N fertilizer dose and allows to alleviate the environmental deterioration related to N pollution. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  13. Heavy metal induced oxidative damage and root morphology alterations of maize (Zea mays L.) plants and stress mitigation by metal tolerant nitrogen fixing Azotobacter chroococcum. (United States)

    Rizvi, Asfa; Khan, Mohd Saghir


    Heavy metals are one of the major abiotic stresses that adversely affect the quantity and nutritive value of maize. Microbial management involving the use of plant growth promoting rhizobacteria (PGPR) is a promising inexpensive strategy for metal clean up from polluted soils. Considering these, metal tolerant plant growth promoting nitrogen fixing rhizobacterial strain CAZ3 identified by 16SrRNA gene sequence analysis as Azotobacter chroococcum was recovered from metal polluted chilli rhizosphere. When exposed to varying levels of metals, A. chroococcum survived up to 1400 and 2000 µg mL -1 of Cu and Pb, respectively and expressed numerous plant growth promoting activities even under metal stress. Strain CAZ3 secreted 65.5 and 60.8 µg mL -1 IAA at 400 µg mL -1 each of Cu and Pb, respectively and produced siderophores, ammonia and ACC deaminase under metal pressure. The melanin extracted from A. chroococcum revealed metal chelating ability under EDX. Following application, strain CAZ3 enhanced growth and yield of maize grown both in the presence of Cu and Pb. The dry biomass of roots of inoculated plants grown with 2007 mg Cu kg -1 and 585 mg Pb kg -1 was increased by 28% and 20%, respectively. At 585 mg Pb kg -1 , the bioinoculant also increased the kernel attributes. At 2007 mg Cu kg -1 strain CAZ3 enhanced the number, yield and protein of kernels by 10%, 45% and 6%, respectively. Interestingly, strain CAZ3 significantly reduced the levels of proline, malondialdehyde and antioxidant enzymes in foliage. The roots of inoculated plants accumulated greatest amounts of metals compared to other organs. In kernels, the concentration of Pb was more as compared to Cu. The metal concentrations in roots, shoots and kernels, however, declined following CAZ3 inoculation. Copper and lead had substantial distortive impact on root and leaf morphology while cell death were visible under CLSM and SEM. Conclusively, A. chroococcum CAZ3 could be a most

  14. Optimising carbon and nitrogen sources for Azotobacter ...

    African Journals Online (AJOL)



    Apr 11, 2011 ... The present work deals with selecting and optimization of carbon and nitrogen sources for producing biomass from Azotobacter chroococcum. Four carbon sources (glucose, sucrose, manitol and sodium benzoate) and four nitrogen sources (yeast extract, meat extract, NH4Cl and (NH4)2SO4) were ...

  15. Optimising carbon and nitrogen sources for Azotobacter ...

    African Journals Online (AJOL)

    The present work deals with selecting and optimization of carbon and nitrogen sources for producing biomass from Azotobacter chroococcum. Four carbon sources (glucose, sucrose, manitol and sodium benzoate) and four nitrogen sources (yeast extract, meat extract, NH4Cl and (NH4)2SO4) were evaluated during the first ...

  16. Effect of Azotobacter croococcum on productive traits and microorganisms in sugar beet rhizosphere

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    Kuzevski Janja


    Full Text Available The aim of this study was to determine the effects of three different inoculation methods with selected Azotobacter chroococcum strains on productive and technological traits of sugar beet, as well as on the total number of microorganisms and azotobacter in rhizosphere. The results of this two-year study showed that effectiveness of the tested inoculation methods in increasing root yield and sugar content varies greatly, depending on year and azotobacter strains. Effectiveness of inoculation methods was not largely impacted by year on granulated sugar. Achieved granulated sugar yield was significantly higher by using pre-sowing azotobacter application, than by using seed inoculation. A significantly increased number of microorganisms in sugar beet rhizosphere was determined, not only by using pre-sowing azotobacter application but also by using sugar beet seed inoculation. Pre-sowing azotobacter application and inter-row cultivation both caused an equal increase in the number of these bacteria in sugar beet rhizosphere (42.2% and 46.9%. Use of sugar beet seed inoculation caused an increase of 33.7% in the number of azotobacter. In order to achieve higher effectiveness in applying azotobacter on productive and technological traits of sugar beet, and considering determined interaction between a certain year, an inoculation method and a strain, it is necessary for future research to focus on determining efficiency of these strains when they are in a mixture.

  17. Effect of two organophosphorus insecticides on the growth, respiration and (14C)-glucose metabolism of Azobacter chroococcum Beij

    International Nuclear Information System (INIS)

    Balasubramanian, A.; Narayanan, R.


    The two organophosphorus insecticides, commonly applied to soil, viz., disulfoton (0,0-diethyl S-2-ethyl thio ethyl phosphorodithioate) and fensulfothion (0,0-diethyl 0-4-methyl sulphinyl phenyl phosphorothioate) did not affect the in vitro growth of Azotobacter chroococcum Beij., the free-living, nitrogen fixing soil bacterium, at 2 ppm (lower level), while the normal dose (5 ppm) and the higher level (10 ppm) suppressed the growth. Respiration of the organism (glucose oxidation) was adversely affected by the insecticides in the growth medium and the inhibition increased with the concentration of the chemical. Both the insecticides suppressed the assimilation of ( 14 C)-glucose in the cold-TCA soluble, hot-TCA soluble fractions and insoluble residue of the cells whereas the 14 C-incorporation in the alcohol soluble and alcohol-ether soluble fractions was enhanced indicating that the insecticides considerably altered the glucose metabolism of the bacterium. (author)


    NARCIS (Netherlands)



    Partial sequences of the dihydrolipoyl transacetylase component (E2p) of the pyruvate dehydrogenase complex from Azotobacter vinelandii and Escherichia coli, containing the catalytic domain, were cloned in pUC plasmids and over-expressed in E. coli TG2. A high expression of a homogeneous protein was


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    A. Khanafari , A. Akhavan Sepahei, M. Mogharab


    Full Text Available Three strains of Azotobacter chroococcum were studied to produce poly-β hydroxybutyrate as a inclusion body by whey degradation. Optimum degradation whey results were obtained when using whey broth as a fermentation medium without extra salt, temperature at 35 °C and pH 7 (P<0.05. Lambda max for whey broth medium was determined probably about 400 nm. The effect of different nitrogenous rich compounds (NH4NO3, Bactopeptone, Casein, Yeast extract, Meat extract, Protease peptone and Tryptone on whey degradation showed that incorporation of nitrogenous compounds into the medium did not increase whey degradation by Azotobacter chroococcum 1723 (P<0.05. But poly-β hydroxyl-butyrate production was increased in presence Meat extract up to 75% of the cell dry weight after 48h. The addition of nitrogenous sourced (except ammonium nitrate had a positive effect on poly-β hydroxyl-butyrate production as it peaked in the presence of Meat extract and 4.43 g/L was accumulated in comparison to 0.5g at diazotrophically growing cells. Increasing the O2 values resulted by shaking at 122 rpm in decreased poly-β hydroxyl-butyrate yield form 4.43 to 0.04 g/L. The results show that this medium supports the growth of strain 1735 and also that this waste could be utilized as a carbon and nitrogen source. Production of poly-β hydroxyl-butyrate by using whey as a medium looks promising, since the use of inexpensive feed-stocks for poly-β hydroxyl-butyrate is essential if bioplastics are to become competitive products.

  20. Impact of Azotobacter exopolysaccharides on sustainable agriculture. (United States)

    Gauri, Samiran S; Mandal, Santi M; Pati, Bikas R


    Recently, increasing attention have lead to search other avenue of biofertilizers with multipurpose activities as a manner of sustainable soil health to improve the plant productivity. Azotobacter have been universally accepted as a major inoculum used in biofertilizer to restore the nitrogen level into cultivated field. Azotobacter is well characterized for their profuse production of exopolysaccharides (EPS). Several reviews on biogenesis and multifunctional role of Azotobacter EPS have been documented with special emphasis on industrial applications. But the impact of Azotobacter EPS in plant growth promotion has not received adequate attention. This review outlines the evidence that demonstrates not only the contribution of Azotobacter EPS in global nutrient cycle but also help to compete successfully in different adverse ecological and edaphic conditions. This also focuses on new insights and concepts of Azotobacter EPS which have positive effects caused by the biofilm formation on overall plant growth promotion with other PGPRs. In addition, their potentials in agricultural improvement are also discussed. Recent data realized that Azotobacter EPS have an immense agro-economical importance including the survivability and maintenance of microbial community in their habitat. This leads us to confirm that the next generation Azotobacter inoculum with high yielding EPS and high nitrogen fixing ability can be utilized to satisfy the future demand of augmented crop production attributed to increase plant growth promoting agents.

  1. Molecular characterization of Azotobacter spp. nifH gene Isolated ...

    African Journals Online (AJOL)

    The nifH gene sequence of the nitrogen-fixing bacterium Azotobacter spp. was determined with the use of polymerase chain reaction (PCR). The Azotobacter species was isolated from marine source in two different seasons. They were cultivated under laboratory conditions using Nitrogen free Azotobacter specific medium.

  2. Molecular characterization of Azotobacter spp. nifH gene Isolated ...

    African Journals Online (AJOL)



    Dec 15, 2009 ... The nifH gene sequence of the nitrogen-fixing bacterium Azotobacter spp. was determined with the use of polymerase chain reaction (PCR). The Azotobacter species was isolated from marine source in two different seasons. They were cultivated under laboratory conditions using Nitrogen free Azotobacter.

  3. Characterization of free nitrogen fixing bacteria of the genus Azotobacter in organic vegetable-grown Colombian soils. (United States)

    Jiménez, Diego Javier; Montaña, José Salvador; Martínez, María Mercedes


    With the purpose of isolating and characterizing free nitrogen fixing bacteria (FNFB) of the genus Azotobacter, soil samples were collected randomly from different vegetable organic cultures with neutral pH in different zones of Boyacá-Colombia. Isolations were done in selective free nitrogen Ashby-Sucrose agar obtaining a recovery of 40%. Twenty four isolates were evaluated for colony and cellular morphology, pigment production and metabolic activities. Molecular characterization was carried out using amplified ribosomal DNA restriction analysis (ARDRA). After digestion of 16S rDNA Y1-Y3 PCR products (1487pb) with AluI, HpaII and RsaI endonucleases, a polymorphism of 16% was obtained. Cluster analysis showed three main groups based on DNA fingerprints. Comparison between ribotypes generated by isolates and in silico restriction of 16S rDNA partial sequences with same restriction enzymes was done with Gen Workbench v.2.2.4 software. Nevertheless, Y1-Y2 PCR products were analysed using BLASTn. Isolate C5T from tomato (Lycopersicon esculentum) grown soils presented the same in silico restriction patterns with A. chroococcum (AY353708) and 99% of similarity with the same sequence. Isolate C5CO from cauliflower (Brassica oleracea var. botrytis) grown soils showed black pigmentation in Ashby-Benzoate agar and high similarity (91%) with A. nigricans (AB175651) sequence. In this work we demonstrated the utility of molecular techniques and bioinformatics tools as a support to conventional techniques in characterization of the genus Azotobacter from vegetable-grown soils.

  4. Characterization of free nitrogen fixing bacteria of the genus Azotobacter in organic vegetable-grown Colombian soils

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    Diego Javier Jiménez


    Full Text Available With the purpose of isolating and characterizing free nitrogen fixing bacteria (FNFB of the genus Azotobacter, soil samples were collected randomly from different vegetable organic cultures with neutral pH in different zones of Boyacá-Colombia. Isolations were done in selective free nitrogen Ashby-Sucrose agar obtaining a recovery of 40%. Twenty four isolates were evaluated for colony and cellular morphology, pigment production and metabolic activities. Molecular characterization was carried out using amplified ribosomal DNA restriction analysis (ARDRA. After digestion of 16S rDNA Y1-Y3 PCR products (1487pb with AluI, HpaII and RsaI endonucleases, a polymorphism of 16% was obtained. Cluster analysis showed three main groups based on DNA fingerprints. Comparison between ribotypes generated by isolates and in silico restriction of 16S rDNA partial sequences with same restriction enzymes was done with Gen Workbench v.2.2.4 software. Nevertheless, Y1-Y2 PCR products were analysed using BLASTn. Isolate C5T from tomato (Lycopersicon esculentum grown soils presented the same in silico restriction patterns with A. chroococcum (AY353708 and 99% of similarity with the same sequence. Isolate C5CO from cauliflower (Brassica oleracea var. botrytis grown soils showed black pigmentation in Ashby-Benzoate agar and high similarity (91% with A. nigricans (AB175651 sequence. In this work we demonstrated the utility of molecular techniques and bioinformatics tools as a support to conventional techniques in characterization of the genus Azotobacter from vegetable-grown soils.

  5. Effect of Selected Azotobacter Bacterial Strains on the Enrichment of ...

    African Journals Online (AJOL)

    Target Audience: Local farmers, Livestock researchers, microbiologist. The effect of three different strains of Azotobacter bacteria in solid substrate fermentation on cassava waste was evaluated. The substrate was incubated at 300c for 10 days inoculation with the Azotobacter bacteria species. One non-inoculated batch ...

  6. Stimulation of Agrobacterium tumefaciens Growth by Azotobacter vinelandii Ferrisiderophores


    Page, William J.; Dale, Phyllis L.


    Azotobacter vinelandii stimulated the growth of Agrobacterium tumefaciens H2, H23, H24, H27, and ATCC 15955 on media containing insoluble iron sources. The Azotobacter vinelandii siderophores appeared to promote Agrobacterium tumefaciens growth by solubilizing mineral iron, and the ferrisiderophores so formed then acted as iron sources for Agrobacterium tumefaciens. Agrobactin, the Agrobacterium siderophore, appeared to be inefficient in solubilizing mineral iron directly.

  7. Comparative study of impact of Azotobacter and Trichoderma with other fertilizers on maize growth

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    Sanjay Mahato


    Full Text Available Biofertilizers may be a better eco-friendly option to maintain soil fertility. The study was conducted to investigate the effect of Azotobacter and Trichoderma on the vegetative growth of maize (Zea mays L. plants. The experiment was carried out in medium sized pots, at IAAS, Lamjung (Feb 2017 - May 2017 in completely randomized design (CRD, consisting eight treatments and three replications. Treatments were namely T1 (control, T2 (Azotobacter, T3 (Trichoderma, T4 (Azotobacter + Trichoderma, T5 (NPK, T6 (Azotobacter + Trichoderma + FYM, T7 (Azotobacter + Trichoderma + FYM + NPK, T8 (FYM. Azotobacter showed a positive increase in plant height, stem girth, dry shoot weight, root length and width, and root weight while Trichoderma displayed either negative or minimal impact. Effect of FYM was lower than Azotobacter but considerably higher than Trichoderma. Trichoderma seriously inhibited the expression of Azotobacter when used together. Trichoderma even suppressed the outcome (except shoot weight of FYM when used together. Root length was the longest in Azotobacter inoculation. The highest number of leaves was in T7 followed by Azotobacter (T2 and NPK (T5. Unlike leaf width, Azotobacter showed a negligible increase in leaves length while Trichoderma wherever present showed the negative impact. Minimum chlorophyll content was found in Azotobacter or Trichoderma after 73 days. Azotobacter treatment showed early tasseling than Trichoderma. The association of Azotobacter and Trichoderma increased the biomass. Azotobacter has significant effects on growth parameters of maize and can supplement chemical fertilizer, while Trichoderma was found to inhibit most of the growth parameters.

  8. Bioremediation of crude oil waste contaminated soil using petrophilic consortium and Azotobacter sp.

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    M. Fauzi


    Full Text Available This study was aimed to determine the effect Petrophilic and Azotobacter sp. consortium on the rate of degradation of hydrocarbons, Azotobacter growth, and Petrophilic fungi growth in an Inceptisol contaminated with crude oil waste originating from Balongan refinery, one of Pertamina (Indonesia’s largest state-owned oil and gas company units in Indramayu – West Java. This study was conducted from March to April 2014 in the glasshouse of research station of the Faculty of Agriculture, Padjadjaran University at Ciparanje, Jatinangor District, Sumedang Regency of West Java. This study used a factorial completely randomized design with two treatments. The first treatment factor was Petrophilic microbes (A consisting of four levels (without treatment, 2% Petrophilic fungi, 2% Petrophilic bacteria, and the 2% Petrophilic consortium, and Azotobacter sp. The second treatment factor was Azotobacter sp. (B consisting of four levels (without treatment, 0.5%, Azotobacter sp., 1% Azotobacter sp., and 1.5% Azotobacter sp. The results demonstrated interaction between Petrophilic microbes and Azotobacter sp. towards hydrocarbon degradation rate, but no interaction was found towards the growth rate of Azotobacter sp. and Petrophilic fungi. Treatments of a1b3 (2% consortium of Petrophilic fungi with 1.5% Azotobacter sp. and a3b3 (2% Petrophilic consortium and 1.5% Azotobacter sp. had hydrocarbon degradation rate at 0.22 ppm/day for each treatment, showing the highest hydrocarbon degradation rate.

  9. Production of extra-cellular polymer in Azotobacter and biosorption ...

    African Journals Online (AJOL)

    Two Azotobacter strains were isolated from alkaline and acid soils. The production of alginate and exopolymer from these two strains showed that, strain AC2 produced high polymer in 2% beet molasses or 1% sucrose broth and addition of nitrogen sources (yeast extract) reduced production of this polymer. The optimum ...


    Tomlinson, Geraldine A.; Campbell, J. J. R.


    Tomlinson, Geraldine A. (The University of British Columbia, Vancouver, B.C., Canada), and J. J. R. Campbell. Patterns of oxidative assimilation in strains of Acetobacter and Azotobacter. J. Bacteriol. 86:1165–1172. 1963.—Oxidative assimilation of glucose-U-C14 was studied with washed-cell suspensions of Acetobacter aceti, A. xylinum, Azotobacter vinelandii, and A. agilis. The suggestion that oxidative assimilation is largely the incorporation of endogenously produced ammonia is tenable. A. aceti did not exhibit oxidative assimilation and it did not incorporate ammonia in the presence of glucose, α-ketoglutarate, or pyruvate. A. xylinum, A. vinelandii, and A. agilis incorporated C14 into the nitrogenous fractions of the cell. The level of assimilation into A. xylinum was low due to the accumulation of extracellular cellulose, and the level of assimilation into the Azotobacter was low presumably because of the requirement of energy for nitrogen fixation. The Azotobacter were characterized by the presence of a high level of radioactivity in the cold trichloroacetic acid-soluble pool. None of the organisms accumulated compounds in the supernatant fluid that might be considered pacemakers in glucose oxidation, and this could be a contributing factor in the low level of assimilation. PMID:14086085

  11. Pengaruh penambahan kultur azotobacter pada feses kambing terhadap kualitas media dan produktivitas cacing tanah (Lumbricus rubellus

    Directory of Open Access Journals (Sweden)

    Nur Cholis


    Full Text Available The purposes of this research were to determine the effect of addition of Azotobacter bacterial culture into media of goat faeces on medium quality and earthworm productivity; and also to examine the best dose of Azotobacter bacterial cultures addition. The research material was 800 g earthworm aged 3 months old. The research method was experimental with Completely Randomized Design using 4 treatments and 4 replications. The results show that addition of Azotobacter bacterial culture had a significant effect (P<0.01 on the medium quality and earthworm productivity (coccoon production, the number of juvenils per coccoon, coccoon hatching percentage, the numbers and weight of earthworm. The bacterial culture addition of 350 cc/100 kg goat faeces was found the best. We suggest to follow the study with observation about the effect of the length of fermentation to the medium quality and earthworm productivity. Keywords: Azotobacter, goat faeces, earthworm

  12. Aerobic Hydrogen Production via Nitrogenase in Azotobacter vinelandii CA6 (United States)

    Noar, Jesse; Loveless, Telisa; Navarro-Herrero, José Luis; Olson, Jonathan W.


    The diazotroph Azotobacter vinelandii possesses three distinct nitrogenase isoenzymes, all of which produce molecular hydrogen as a by-product. In batch cultures, A. vinelandii strain CA6, a mutant of strain CA, displays multiple phenotypes distinct from its parent: tolerance to tungstate, impaired growth and molybdate transport, and increased hydrogen evolution. Determining and comparing the genomic sequences of strains CA and CA6 revealed a large deletion in CA6's genome, encompassing genes related to molybdate and iron transport and hydrogen reoxidation. A series of iron uptake analyses and chemostat culture experiments confirmed iron transport impairment and showed that the addition of fixed nitrogen (ammonia) resulted in cessation of hydrogen production. Additional chemostat experiments compared the hydrogen-producing parameters of different strains: in iron-sufficient, tungstate-free conditions, strain CA6's yields were identical to those of a strain lacking only a single hydrogenase gene. However, in the presence of tungstate, CA6 produced several times more hydrogen. A. vinelandii may hold promise for developing a novel strategy for production of hydrogen as an energy compound. PMID:25911479

  13. Screening of Azotobacter isolates for PGP properties and antifungal activity

    Directory of Open Access Journals (Sweden)

    Bjelić Dragana Đ.


    Full Text Available Аmong 50 bacterial isolates obtained from maize rhizospherе, 13 isolates belonged to the genus Azotobacter. Isolates were biochemically characterized and estimated for pH and halo tolerance ability and antibiotic resistance. According to characterization, the six representative isolates were selected and further screened in vitro for plant growth promoting properties: production of indole-3-acetic acid (IAA, siderophores, hydrogen cyanide (HCN, exopolysaccharides, phosphate solubilization and antifungal activity (vs. Helminthosporium sp., Macrophomina sp., Fusarium sp.. Beside HCN production, PGP properties were detected for all isolates except Azt7. All isolates produced IAA in the medium without L-tryptophan and the amount of produced IAA increased with concentration of precursor in medium. The highest amount of IAA was produced by isolates Azt4 (37.69 and 45.86 μg ml-1 and Azt5 (29.44 and 50.38 μg ml-1 in the medium with addition of L-tryptophan (2.5 and 5 mM. The isolates showed the highest antifungal activity against Helminthosporium sp. and the smallest antagonistic effect on Macrophomina sp. Radial Growth Inhibition (RGI obtained by the confrontation of isolates with tested phytopathogenic fungi, ranged from 10 to 48%. [Projekat Ministarstva nauke Republike Srbije, br. TR 31073

  14. The gram-negative bacterium Azotobacter chroococcum NCIMB 8003 employs a new glycoside hydrolase family 70 4,6-α-glucanotransferase enzyme (GtfD) to synthesize a reuteran like polymer from maltodextrins and starch

    NARCIS (Netherlands)

    Gangoiti, Joana; van Leeuwen, Sander S; Vafiadi, Christina; Dijkhuizen, Lubbert

    BACKGROUND: Originally the glycoside hydrolase (GH) family 70 only comprised glucansucrases of lactic acid bacteria which synthesize α-glucan polymers from sucrose. Recently we have identified 2 novel subfamilies of GH70 enzymes represented by the Lactobacillus reuteri 121 GtfB and the

  15. Encystment of Azotobacter nigricans grown diazotrophically on kerosene as sole carbon source. (United States)

    García-Esquivel, Gabriela; Calva-Calva, Graciano; Ferrera-Cerrato, Ronald; Fernández-Linares, Luis Carlos; Vázquez, Refugio Rodríguez; Esparza-García, Fernando José


    Encystment of Azotobacter nigricans was induced by its diazotrophic cultivation on kerosene. Its growth and nitrogenase activity were affected by kerosene in comparison to cultures grown on sucrose. Electron microscopy of vegetative cells showed that when nitrogenase activity was higher and the poly-beta-hydroxybutyrate granules were not present to a significant extent, peripheral bodies were abundant. After 8 days of culture on kerosene, the presence of cysts with intracellular bunches of poly-beta-hydroxybutyrate granules was observed. Germination of cysts bears germinating multicelled yet unbroken capsule cysts with up to three cells inside. This is the first report of encystment induction of Azotobacter species grown on kerosene.

  16. Growth of Avicennia marina and Ceriops decandra seedlings inoculated with halophilic azotobacters. (United States)

    Ravikumar, S; Kathiresan, K; Alikhan, S Liakath; Williams, G Prakash; Gracelin, N Anitha Anandha


    Inoculation of azotobacter has significant positive effects on the growth characteristics and pigments in mangrove seedlings of Avicennia marina and Ceriops decandra. The bacterial inoculation significantly increased the root dry biomass at the maximum of 75.8% at 30 gl(-1) salinity in Ceriops decandra. But in Avicennia marina, the shoot dry biomass was increased significantly at the maximum of 56.12% at 30 gl(-1) salinity in general, the Azotobacter beijerinkii improved the growth characteristics better in both species of mangroves preferably at higher salinity levels in A. marina and at a range of salinity in C. decandra. The results recommend this forraising vigorous seedlings under nursery conditions.

  17. Potential of Azotobacter vinelandii Khsr1 as bio-inoculant | Naz ...

    African Journals Online (AJOL)

    The present study deals with the isolation and characterization of Azotobacter vinelandii Khsr1 from roots of the weed, Chrysopogon aucheri, commonly known as golden beard grass indigenous to Khewra salt range, Pakistan and its evaluation as bio-inoculant. The population of the isolate varied from 107 to 108 cfu/g fresh ...

  18. Defining the Pseudomonas Genus: Where Do We Draw the Line with Azotobacter?

    DEFF Research Database (Denmark)

    Özen, Asli Ismihan; Ussery, David


    The genus Pseudomonas has gone through many taxonomic revisions over the past 100 years, going from a very large and diverse group of bacteria to a smaller, more refined and ordered list having specific properties. The relationship of the Pseudomonas genus to Azotobacter vinelandii is examined...

  19. Differential accumulation of nif structural gene mRNA in Azotobacter vinelandii. (United States)

    Hamilton, Trinity L; Jacobson, Marty; Ludwig, Marcus; Boyd, Eric S; Bryant, Donald A; Dean, Dennis R; Peters, John W


    Northern analysis was employed to investigate mRNA produced by mutant strains of Azotobacter vinelandii with defined deletions in the nif structural genes and in the intergenic noncoding regions. The results indicate that intergenic RNA secondary structures effect the differential accumulation of transcripts, supporting the high Fe protein-to-MoFe protein ratio required for optimal diazotrophic growth.

  20. Formation of on- and off-pathway intermediates in the folding kinetics of Azotobacter vinelandii apoflavodoxin.

    NARCIS (Netherlands)

    Bollen, Y.J.M.; Sanchez, I.E.; Mierlo, van C.P.M.


    The folding kinetics of the 179-residue Azotobacter vinelandii apoflavodoxin, which has an alpha-beta parallel topology, have been followed by stopped-flow experiments monitored by fluorescence intensity and anisotropy. Single-jump and interrupted refolding experiments show that the refolding

  1. Formation of on- and off-pathway intermediates in the folding kinetics of azotobacter vinelandii apoflavodoxin.

    NARCIS (Netherlands)

    Bollen, Y.J.M.; Sanchez, I.E.; Van Mierlo, C.P.M.


    The folding kinetics of the 179-residue Azotobacter vinelandii apoflavodoxin, which has an α-β parallel topology, have been followed by stopped-flow experiments monitored by fluorescence intensity and anisotropy. Single-jump and interrupted refolding experiments show that the refolding kinetics

  2. Energy supply for dinitrogen fixation by Azotobacter vinelandii and by bacteroids of Rhizobium leguminosarum

    NARCIS (Netherlands)

    Laane, N.C.M.


    The central issue of this thesis is how obligate aerobes, such as Rhizobium leguminosarum bacteroids and Azotobacter vinelandii, generate and regulate the energy supply (in the form of ATP and reducing equivalents) for nitrogenase.
    In an effective

  3. Catalytic mechanism of hydrogenase from Azotobacter vinelandii. Final technical report, August 1, 1994--July 31, 1997

    Energy Technology Data Exchange (ETDEWEB)

    Arp, D.J.


    This project is focused on investigations of the catalytic mechanism of the hydrogenase found in the aerobic, N{sub 2}-fixing microorganism Azotobacter vinelandii. This report summarizes the progress during the first two years of the current project and include the anticipated course of the research for the remaining year of the current project. Because the current proposal represents a change in direction, the authors also include a brief progress report of prior DOE-sponsored research dealing with hydrogenases.



    Malynovska I. M.


    The aim of the work was to determine the possibility of using the number and activity of Azotobacter cells and melanin-synthesizing micromycetes as indicators of gray forest soils of different types (fallow, extensive and intensive agrosoil) pollution with heavy metal ions. For this purpose, there were used laboratory-analytical, microbiological and statistical methods. As a result of research of increasing doses of heavy metals (zinc + lead) influence on the number of microorganisms in the g...

  5. Metabolism of resorcinylic compounds by bacteria: new pathway for resorcinol catabolism in Azotobacter vinelandii.


    Groseclose, E E; Ribbons, D W


    We present evidence to document a third pathway for the microbial catabolism of resorcinol. Resorcinol is converted to pyrogallol by resorcinol-grown cells of Azotobacter vinelandii. Pyrogallol is the substrate for one of two ring cleavage enzymes induced by growth with resorcinol. Oxalocrotonate, CO2, pyruvate, and acetaldehyde have been identified as products of pyrogallol oxidation catalyzed by extracts of resorcinol-grown cells. The enzymes pyrogallol 1,2-dioxygenase, oxalocrotonate tauto...

  6. Physical and genetic map of the major nif gene cluster from Azotobacter vinelandii.


    Jacobson, M R; Brigle, K E; Bennett, L T; Setterquist, R A; Wilson, M S; Cash, V L; Beynon, J; Newton, W E; Dean, D R


    Determination of a 28,793-base-pair DNA sequence of a region from the Azotobacter vinelandii genome that includes and flanks the nitrogenase structural gene region was completed. This information was used to revise the previously proposed organization of the major nif cluster. The major nif cluster from A. vinelandii encodes 15 nif-specific genes whose products bear significant structural identity to the corresponding nif-specific gene products from Klebsiella pneumoniae. These genes include ...

  7. Effects of macro nutrient concentration on biological N2 fixation by Azotobacter vinelandii ATCC 12837

    International Nuclear Information System (INIS)

    Liew Pauline Woan Ying; Nazalan Najimudin; Jong Bor Chyan; Latiffah Noordin; Khairuddin Abdul Rahim; Amir Hamzah Ahmad Ghazali


    The dynamic changes of biological N 2 fixation by Azotobacter vinelandii ATCC 12837 under the influence of various macro nutrients, specifically phosphorus (P) and potassium (K), was investigated. In this attempt, Oryza sativa L. var. MR 219 was used as the model plant. Results obtained showed changes in the biological N 2 fixation activities with different macro nutrient(s) manipulations. The research activity enables optimisation of macro nutrients concentration for optimal/ enhanced biological N 2 fixation by A. vinelandii ATCC 12837. (author)


    Directory of Open Access Journals (Sweden)

    Malynovska I. M.


    Full Text Available The aim of the work was to determine the possibility of using the number and activity of Azotobacter cells and melanin-synthesizing micromycetes as indicators of gray forest soils of different types (fallow, extensive and intensive agrosoil pollution with heavy metal ions. For this purpose, there were used laboratory-analytical, microbiological and statistical methods. As a result of research of increasing doses of heavy metals (zinc + lead influence on the number of microorganisms in the gray forest soils it was found that the number and activity of Azotobacter and the number and part of melanin-synthesizing micromycetes in their total number may be fit into indicators of pollution with heavy metals. Azotobacter cells activity index may be considered indicative at contamination levels of 5-100 of maximum permissible concentration in the absence of vegetation, at contamination levels of 10–100 – for soils with phytocenosis. The number and proportion of melaninsynthesizing micromycetes in total guantity may serve as diagnostic sign of gray forest soils pollution with high doses of heavy metals, but only for the period of contamination up to 2 years. It was shown that nature of the effect of heavy metals on the number of microorganisms of indicative groups depended on the presence of plants in the monitoring system, on doses of heavy metals, on the term of contamination and on the type of soil usage.

  9. Hamiltonian purification

    Energy Technology Data Exchange (ETDEWEB)

    Orsucci, Davide [Scuola Normale Superiore, I-56126 Pisa (Italy); Burgarth, Daniel [Department of Mathematics, Aberystwyth University, Aberystwyth SY23 3BZ (United Kingdom); Facchi, Paolo; Pascazio, Saverio [Dipartimento di Fisica and MECENAS, Università di Bari, I-70126 Bari (Italy); INFN, Sezione di Bari, I-70126 Bari (Italy); Nakazato, Hiromichi; Yuasa, Kazuya [Department of Physics, Waseda University, Tokyo 169-8555 (Japan); Giovannetti, Vittorio [NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, I-56126 Pisa (Italy)


    The problem of Hamiltonian purification introduced by Burgarth et al. [Nat. Commun. 5, 5173 (2014)] is formalized and discussed. Specifically, given a set of non-commuting Hamiltonians (h{sub 1}, …, h{sub m}) operating on a d-dimensional quantum system ℋ{sub d}, the problem consists in identifying a set of commuting Hamiltonians (H{sub 1}, …, H{sub m}) operating on a larger d{sub E}-dimensional system ℋ{sub d{sub E}} which embeds ℋ{sub d} as a proper subspace, such that h{sub j} = PH{sub j}P with P being the projection which allows one to recover ℋ{sub d} from ℋ{sub d{sub E}}. The notions of spanning-set purification and generator purification of an algebra are also introduced and optimal solutions for u(d) are provided.

  10. Uji Kemampuan Bakteri Azotobacter S8 dan Bacillus subtilis untuk Menyisihkan Trivalent Chromium (Cr3+ pada Limbah Cair

    Directory of Open Access Journals (Sweden)

    Muhammad Fauzul Imron


    Full Text Available Penggunaan kromium dalam berbagai industri telah menyebabkan pencemaran pada lingkungan. Limbah kromium yang sering ditemukan pada badan air dalam bentuk Cr3+ dan Cr6+. Bakteri diketahui mampu menyisihkan logam berat kromium sehingga dapat digunakan sebagai agen bioremediasi. Azotobacter S8 dan Bacillus subtilis merupakan bakteri yang mampu menyisihkan logam berat kromium. Tujuan dari penelitian ini adalah untuk menentukan komposisi optimum dan persentase penyisihan logam berat kromium oleh bakteri Azotobacter S8 dan Bacillus subtilis baik secara tunggal ataupun konsortium. Hasil penelitian ini menunjukkan bahwa persentase penyisihan tertinggi dilakukan oleh bakteri tunggal Azotobacter S8 yaitu 10,53% pada konsentrasi 50 mg/L dengan waktu 4 jam dan pH 8,35 serta jumlah koloni akhir yang terukur adalah 4 x 109 CFU/ml.

  11. [Isolation and characterization of Azotobacter sp. for the production of poly-beta-hydroxyalkanoates]. (United States)

    Quagliano, J C; Alegre, P; Miyazaki, S S


    A microorganism (Azotobacter sp.) was isolated from soil samples from the Agronomy Faculty campus and its ability to accumulate poly-beta-hydroxyalkanoate (PHAs) polymers was analyzed. The isolated strain (named No 8) accumulated 8 micrograms PHA/m of culture media as intracytoplasmic granules. The following properties of the strain were analyzed: utilization of different carbon sources, antibiotic resistance, optimal pH and temperature, pigment production, nitrogen fixation, proteolytic activity, acid and hydrogen sulphide production, optimal growth temperature, cyst formation, growth on phenol and sodium fluoride, catalase, pleomorfism and mobility. The synthesized polymer showed valuable properties respect to organic solvents resistance.

  12. Batch culture of Azotobacter vinelandii under oxygen limitation conditionS

    Energy Technology Data Exchange (ETDEWEB)

    Camacho Rubio, F.; Martinez Nieto, L.; Fernandez Serrano, M.; Jimenez Moleon, M.C. [Departamento de Ingenieria Quimica, Universidad de Granada, Granada (Spain)


    The batch culture of Azotobacter vinealandii on glucose under nitrogen-fixing conditions, seeking oxygen limitation conditions, has been studied in order to use it as a Biological Test System for the experimental study of oxygen transfer enhancement methods in aerobic fermenters. overall kinetic parameters for exponential growth and for linear growth (under oxygen limitation) have been determined. It was noted an appreciable influence of the oxygen transfer rate on glucose and oxygen uptake, which seems to be due to alginate production, excreted as a nitrogenase protection mechanisms. (Author) 12 refs.

  13. Alginate Biosynthesis in Azotobacter vinelandii: Overview of Molecular Mechanisms in Connection with the Oxygen Availability

    Directory of Open Access Journals (Sweden)

    Ivette Pacheco-Leyva


    Full Text Available The Gram-negative bacterium Azotobacter vinelandii can synthetize the biopolymer alginate that has material properties appropriate for plenty of applications in industry as well as in medicine. In order to settle the foundation for improving alginate production without compromising its quality, a better understanding of the polymer biosynthesis and the mechanism of regulation during fermentation processes is necessary. This knowledge is crucial for the development of novel production strategies. Here, we highlight the key aspects of alginate biosynthesis that can lead to producing an alginate with specific material properties with particular focus on the role of oxygen availability linked with the molecular mechanisms involved in the alginate production.

  14. Pengaruh pupuk kandang dan NPK terhadap populasi bakteri Azotobacter dan Azospirillum dalam tanah pada budidaya cabai (Capsicum annum

    Directory of Open Access Journals (Sweden)



    Full Text Available Mujiyati, Supriyadi. 2009. Pengaruh pupuk kandang dan NPK terhadap populasibakteri Azotobacter dan Azospirillum dalam tanah pada budidaya cabai (Capsicumannum. Bioteknologi 6: 63-69. Tujuan penelitian ini adalah mengetahui peningkatan populasi bakteri Azotobacter dan Azospirillum akibat pemberian pupuk kandang. Percobaan menggunakan rancangan acak kelompok dengan perlakuan: (i tanpa pupuk sebagai kontrol, (ii dengan pupuk kandang, (iii dengan pupuk NPK. Data dikumpulkan secara eksperimen dengan menanam cabai pada beberapa petak percobaan dengan perlakuan penggunaan pupuk sebanyak tiga kali ulangan. Data yang dikumpulkan terdiri atas jumlah populasi Azotobacter dan Azospirillum, kandungan nitrogen dalam tanah dan hasil cabai. Percobaan lapangan dilakukan di Desa Gathak, Kecamatan Karangnongko, Kabupaten Klaten, Jawa Tengah. Data primer dari hasil penelitian di analisis dengan uji ANAVA dan dilanjutkan dengan uji BNT dengan tingkat kepercayaan 95%. Hasil penelitian menunjukkan bahwa menggunakan pupuk kandang dapat meningkatkan populasi bakteri Azotobacter (0,02% dan Azospirillum (0,46% apabila dibandingkan kontrol, sehingga dapat meningkatkan kesuburan tanah dalam waktu yang cukup lama yaitu dengan meningkatkan ketersediaan hara dalam tanah. Kandungan nitrogen total tanah setelah diberi pupuk kandang juga meningkat dan sangat bermanfaat untuk bahan penyusun tubuh tumbuhan.

  15. Protein quantity and quality of safflower seed improved by NP fertilizer and rhizobacteria (Azospirillum and Azotobacter spp.

    Directory of Open Access Journals (Sweden)

    Asia eNosheen


    Full Text Available Protein is an essential part of human diet. The aim of present study was to improve the protein quality of safflower seed by the application of plant growth promoting rhizobacteria (PGPR in combination with conventional nitrogen and phosphate (NP fertilizers. The seeds of two safflower cultivar Thori and Saif-32, were inoculated with Azospirillum and Azotobacter and grown under field conditions. Protein content and quality was assessed by crude protein, amino acid analysis and SDS-PAGE. Seed crude protein and amino acids (metheonine, phenylanine and glutamic acid showed significant improvement (55%–1250% by Azotobacter supplemented with quarter dose of fertilizers (BTQ at P≤0.05. Additional protein bands were induced in Thori and Saif-32 by BTQ and BTH (Azotobacter supplemented with half dose of fertilizers respectively. The Azospirillum in combination with half dose of fertilizers (SPH and BTQ enhanced the indole acetic acid (90% and gibberellic acid (23%–27% contents in safflower leaf. Taken together, these data suggest that Azospirillum and Azotobacter along with significantly reduced (up to 75% use of NP fertilizers improved the quality and quantity of safflower seed protein.

  16. Characterization of free nitrogen fixing bacteria of the genus Azotobacter in organic vegetable-grown Colombian soils

    NARCIS (Netherlands)

    Jiménez Avella, Diego; Montaña, José Salvador; Martínez, María Mercedes

    With the purpose of isolating and characterizing free nitrogen fixing bacteria (FNFB) of the genus Azotobacter, soil samples were collected randomly from different vegetable organic cultures with neutral pH in different zones of Boyacá-Colombia. Isolations were done in selective free nitrogen

  17. Purification and cellular localization of wild type and mutated dihydrolipoyltransacetylases from Azotobacter vinelandii and Escherichia coli expressed in E. coli

    NARCIS (Netherlands)

    Schulze, Egbert; Westphal, Adrie H.; Veenhuis, Marten; Kok, Arie de


    Wild type dihydrolipoyltransacetylase(E2p)-components from the pyruvate dehydrogenase complex of A. vinelandii or E. coli, and mutants of A. vinelandii E2p with stepwise deletions of the lipoyl domains or the alanine- and proline-rich region between the binding and the catalytic domain have been

  18. Cloning of the sth gene from Azotobacter vinelandii and construction of chimeric soluble pyridine nucleotide transhydrogenases. (United States)

    Boonstra, B; Björklund, L; French, C E; Wainwright, I; Bruce, N C


    The gene encoding the soluble pyridine nucleotide transhydrogenase (STH) of Azotobacter vinelandii was cloned and sequenced. This is the third sth gene identified and further defines a new subfamily within the flavoprotein disulfide oxidoreductases. The three STHs identified all lack one of the redox active cysteines that are characteristic for this large family of enzymes, and instead they contain a conserved threonine residue at this position. The recombinant A. vinelandii enzyme was purified to homogeneity and shown to form filamentous structures different from those of Pseudomonas fluorescens and Escherichia coli STH. Chimeric STHs were constructed which showed that the C-terminal region is important for polymer formation. The A. vinelandii STH containing the C-terminal region of P. fluorescens or E. coli STH showed structures resembling those of the STH contributing the C-terminal portion of the protein.

  19. Influence of N-15 labelled urea and azotobacter on corn yield and nitrogen budget as affected by organic matter

    International Nuclear Information System (INIS)

    Soliman, S.M.; Abdelmonem, M.A.


    As sandy soils of Egypt are poor in their chemical and physical properties, their fertilization with chemical or biological fertilizer is essential. The reported greenhouse experiment was conducted, using sandy soil of Egypt to evaluate the impact of urea fertilizer, applied alone combined with nitrification inhibitors (DCD) or (N-serve) on (corn yield and N-losses) as compared with inoculation with azotobacter under organic matter treatment as soil amendment. Total dry matter was recorded, while N-uptake by corn, and N-recovery due to inoculation with azotobacter was determined using N-15 dilution technique. Data obtained indicated that, application of DCD or N-serve with urea increased corn dry matter weight as well N-15 recovery. Significant increase in N-recovery was obtained due to nitrification inhibitor application and azotobacter inoculation. N-15 losses were reduced due to application of DCD and N-serve from 45% and respectively. Use of bio fertilizers and nitrification inhibitors could play an important role in corn production in sandy soil, as well as decrease the losses of applied N-fertilizers. fig., 3 tabs

  20. Phenotypic and molecular characterisation of efficient nitrogen-fixing Azotobacter strains from rice fields for crop improvement. (United States)

    Sahoo, Ranjan K; Ansari, Mohammad W; Dangar, Tushar K; Mohanty, Santanu; Tuteja, Narendra


    Biological nitrogen fixation (BNF) is highly effective in the field and potentially useful to reduce adverse effects chemical fertilisers. Here, Azotobacter species were selected via phenotypic, biochemical and molecular characterisations from different rice fields. Acetylene reduction assay of Azotobacter spp. showed that Azotobacter vinelandii (Az3) fixed higher amount of nitrogen (121.09 nmol C2H4 mg(-1) bacteria h(-1)). Likewise, its plant growth functions, viz. siderophore, hydrogen cyanide, salicylic acid, IAA, GA3, zeatin, NH3, phosphorus solubilisation, ACC deaminase and iron tolerance, were also higher. The profile of gDNA, plasmid DNA and cellular protein profile depicted inter-generic and inter-specific diversity among the isolates of A. vinelandii. The PCR-amplified genes nifH, nifD and nifK of 0.87, 1.4 and 1.5 kb , respectively, were ascertained by Southern blot hybridisation in isolates of A. vinelandii. The 16S rRNA sequence from A. vinelandii (Az3) was novel, and its accession number (JQ796077) was received from NCBI data base. Biofertiliser formulation of novel A. vinelandii isolates along with commercial one was evaluated in rice (Oriza sativa L. var. Khandagiri) fields. The present finding revealed that treatment T4 (Az3) (A. vinelandii) are highly efficient to improved growth and yield of rice crop.

  1. Effects of Chemical Components on the Growth of Azotobacter vinelandii Mutant and PHB Production

    International Nuclear Information System (INIS)

    Nur Izzah Mohd Razak; Safiyyah Zainuddin; Ying, P.L.W.; Chyan, J.B.; Elly Ellyna Rashid


    Polyhydroxy butyrate (PHB) is a non-toxic biodegradable polymer produced by some bacteria and can be applied in medical, pharmacology and food industry. Eight types of chemical components acting as supplements were added to culture medium as nutrient pulses. The growth of bacteria was monitored by recording the absorbance value at 600 nm for every 24 hours of cultivation. PHB was extracted using chloroform. Reading at 235 nm was recorded to determine the PHB concentration. By observing the dry cell weight, we observed the addition of sucrose increased the biomass of Azotobacter vinelandii mutant from 1.5 mg/ mL to 5.6 mg/ mL and 8.4 mg/ mL after 3, 4 and 7 days of cultivation, respectively. The highest PHB concentration of 859.27 μg/ mL was obtained after seven days of cultivation in the medium which was supplied with urea sequentially, With sucrose, the PHB concentration increased from 25.45 μg/ ml to 99.59 μg/ ml and 655.56 μg/ ml after 3, 4 and 7 days of cultivation, respectively. The PHB concentrations obtained with sucrose-pulses were the second highest after urea. As a conclusion, sucrose and urea are the two major factors in the growth and PHB production by A. vinelandii mutant. (author)

  2. Production of polyhydroxybutyrate and alginate from glycerol by Azotobacter vinelandii under nitrogen-free conditions. (United States)

    Yoneyama, Fuminori; Yamamoto, Mayumi; Hashimoto, Wataru; Murata, Kousaku


    Glycerol is an interesting feedstock for biomaterials such as biofuels and bioplastics because of its abundance as a by-product during biodiesel production. Here we demonstrate glycerol metabolism in the nitrogen-fixing species Azotobacter vinelandii through metabolomics and nitrogen-free bacterial production of biopolymers, such as poly-d-3-hydroxybutyrate (PHB) and alginate, from glycerol. Glycerol-3-phosphate was accumulated in A. vinelandii cells grown on glycerol to the exponential phase, and its level drastically decreased in the cells grown to the stationary growth phase. A. vinelandii also overexpressed the glycerol-3-phosphate dehydrogenase gene when it was grown on glycerol. These results indicate that glycerol was first converted to glycerol-3-phosphate by glycerol kinase. Other molecules with industrial interests, such as lactic acid and amino acids including γ-aminobutyric acid, have also been accumulated in the bacterial cells grown on glycerol. Transmission electron microscopy revealed that glycerol-grown A. vinelandii stored PHB within the cells. The PHB production level reached 33% per dry cell weight in nitrogen-free glycerol medium. When grown on glycerol, alginate-overproducing mutants generated through chemical mutagenesis produced 2-fold the amount of alginate from glycerol than the parental wild-type strain. To the best of our knowledge, this is the first report on bacterial production of biopolymers from glycerol without addition of any nitrogen source.

  3. Physiology and biochemistry of polysaccharide production by Azotobacter vinelandii and Pseudomonas aeruginosa

    Energy Technology Data Exchange (ETDEWEB)

    Annison, G.


    The extracellular production and the composition of the acetylated alginates of Azotobacter vinelandii and Pseudomonas aeruginosa were investigated. A reverse-phase HPLC method to determine D-mannuronate/L-guluronate (M/G) ratios in alginates was developed. Comparison between the HPLC procedure and a /sup 1/H-NMR method was made. Growth and alginate production by A. vinelandii in batch and continuous culture were studied. Polysaccharide production was favored in batch culture by reduced aeration rates by by intermediate aeration rates in continuous culture. The partitioning of Ca/sup 45/ in solution and associated with alginates during the growth cycle of A. vinelandii was studied. The change in the composition of polysaccharide produced during growth of A. vinelandii was related to the level of free Ca/sup + +/ which fell rapidly during initial stages of incubation. The degree of acetylation of the polyuronate was shown to be variable. Production of alginates with high O-acetate contents was observed in cultures maintained at high growth rates and high Ca/sup + +/ levels. The degree of acetylation of the polyuronate was not related to the level of epimerization in vivo although in vitro studies demonstrated that chemical acetylation of the alginate inhibited epimerization. Alginate production by clinical isolates of Pseudomonas aeruginosa was shown to be unstable and no polyguluronate was isolated from cultures. It was also noted that Ca/sup + +/ played a less important role in the composition of the polysaccharide.

  4. Gene Deletions Resulting in Increased Nitrogen Release by Azotobacter vinelandii: Application of a Novel Nitrogen Biosensor (United States)

    Eberhart, Lauren J.; Ohlert, Janet M.; Knutson, Carolann M.; Plunkett, Mary H.


    Azotobacter vinelandii is a widely studied model diazotrophic (nitrogen-fixing) bacterium and also an obligate aerobe, differentiating it from many other diazotrophs that require environments low in oxygen for the function of the nitrogenase. As a free-living bacterium, A. vinelandii has evolved enzymes and transporters to minimize the loss of fixed nitrogen to the surrounding environment. In this study, we pursued efforts to target specific enzymes and further developed screens to identify individual colonies of A. vinelandii producing elevated levels of extracellular nitrogen. Targeted deletions were done to convert urea into a terminal product by disrupting the urease genes that influence the ability of A. vinelandii to recycle the urea nitrogen within the cell. Construction of a nitrogen biosensor strain was done to rapidly screen several thousand colonies disrupted by transposon insertional mutagenesis to identify strains with increased extracellular nitrogen production. Several disruptions were identified in the ammonium transporter gene amtB that resulted in the production of sufficient levels of extracellular nitrogen to support the growth of the biosensor strain. Further studies substituting the biosensor strain with the green alga Chlorella sorokiniana confirmed that levels of nitrogen produced were sufficient to support the growth of this organism when the medium was supplemented with sufficient sucrose to support the growth of the A. vinelandii in coculture. The nature and quantities of nitrogen released by urease and amtB disruptions were further compared to strains reported in previous efforts that altered the nifLA regulatory system to produce elevated levels of ammonium. These results reveal alternative approaches that can be used in various combinations to yield new strains that might have further application in biofertilizer schemes. PMID:25888177

  5. Structural Basis for Cyclization Specificity of Two Azotobacter Type III Polyketide Synthases (United States)

    Satou, Ryutaro; Miyanaga, Akimasa; Ozawa, Hiroki; Funa, Nobutaka; Katsuyama, Yohei; Miyazono, Ken-ichi; Tanokura, Masaru; Ohnishi, Yasuo; Horinouchi, Sueharu


    Type III polyketide synthases (PKSs) show diverse cyclization specificity. We previously characterized two Azotobacter type III PKSs (ArsB and ArsC) with different cyclization specificity. ArsB and ArsC, which share a high sequence identity (71%), produce alkylresorcinols and alkylpyrones through aldol condensation and lactonization of the same polyketomethylene intermediate, respectively. Here we identified a key amino acid residue for the cyclization specificity of each enzyme by site-directed mutagenesis. Trp-281 of ArsB corresponded to Gly-284 of ArsC in the amino acid sequence alignment. The ArsB W281G mutant synthesized alkylpyrone but not alkylresorcinol. In contrast, the ArsC G284W mutant synthesized alkylresorcinol with a small amount of alkylpyrone. These results indicate that this amino acid residue (Trp-281 of ArsB or Gly-284 of ArsC) should occupy a critical position for the cyclization specificity of each enzyme. We then determined crystal structures of the wild-type and G284W ArsC proteins at resolutions of 1.76 and 1.99 Å, respectively. Comparison of these two ArsC structures indicates that the G284W substitution brings a steric wall to the active site cavity, resulting in a significant reduction of the cavity volume. We postulate that the polyketomethylene intermediate can be folded to a suitable form for aldol condensation only in such a relatively narrow cavity of ArsC G284W (and presumably ArsB). This is the first report on the alteration of cyclization specificity from lactonization to aldol condensation for a type III PKS. The ArsC G284W structure is significant as it is the first reported structure of a microbial resorcinol synthase. PMID:24100027

  6. Molecular and bioengineering strategies to improve alginate and polydydroxyalkanoate production by Azotobacter vinelandii

    Directory of Open Access Journals (Sweden)

    Espín Guadalupe


    Full Text Available Abstract Several aspects of alginate and PHB synthesis in Azotobacter vinelandii at a molecular level have been elucidated in articles published during the last ten years. It is now clear that alginate and PHB synthesis are under a very complex genetic control. Genetic modification of A. vinelandii has produced a number of very interesting mutants which have particular traits for alginate production. One of these mutants has been shown to produce the alginate with the highest mean molecular mass so far reported. Recent work has also shed light on the factors determining molecular mass distribution; the most important of these being identified as; dissolved oxygen tension and specific growth rate. The use of specific mutants has been very useful for the correct analysis and interpretation of the factors affecting polymerization. Recent scale-up/down work on alginate production has shown that oxygen limitation is crucial for producing alginate of high molecular mass, a condition which is optimized in shake flasks and which can now be reproduced in stirred fermenters. It is clear that the phenotypes of mutants grown on plates are not necessarily reproducible when the strains are tested in lab or bench scale fermenters. In the case of PHB, A. vinelandii has shown itself able to produce relatively large amounts of this polymer of high molecular weight on cheap substrates, even allowing for simple extraction processes. The development of fermentation strategies has also shown promising results in terms of improving productivity. The understanding of the regulatory mechanisms involved in the control of PHB synthesis, and of its metabolic relationships, has increased considerably, making way for new potential strategies for the further improvement of PHB production. Overall, the use of a multidisciplinary approach, integrating molecular and bioengineering aspects is a necessity for optimizing alginate and PHB production in A. vinelandii.

  7. The involvement of the fixABCX genes and the respiratory chain in the electron transport to nitrogenase in Azotobacter vinelandii

    NARCIS (Netherlands)

    Wientjens, R.


    The work in this thesis is mainly focused on the electron transport route to nitrogenase in the free-living, obligate aerobic, nitrogen fixing organism Azotobacter vinelandii. For many years now, this topic has been the subject of

  8. Spectroscopic and functional characterization of iron-bound forms of Azotobacter vinelandii (Nif)IscA. (United States)

    Mapolelo, Daphne T; Zhang, Bo; Naik, Sunil G; Huynh, Boi Hanh; Johnson, Michael K


    The ability of Azotobacter vinelandii(Nif)IscA to bind Fe has been investigated to assess the role of Fe-bound forms in NIF-specific Fe-S cluster biogenesis. (Nif)IscA is shown to bind one Fe(III) or one Fe(II) per homodimer and the spectroscopic and redox properties of both the Fe(III)- and Fe(II)-bound forms have been characterized using the UV-visible absorption, circular dichroism, and variable-temperature magnetic circular dichroism, electron paramagnetic resonance, Mössbauer and resonance Raman spectroscopies. The results reveal a rhombic intermediate-spin (S = 3/2) Fe(III) center (E/D = 0.33, D = 3.5 ± 1.5 cm(-1)) that is most likely 5-coordinate with two or three cysteinate ligands and a rhombic high spin (S = 2) Fe(II) center (E/D = 0.28, D = 7.6 cm(-1)) with properties similar to reduced rubredoxins or rubredoxin variants with three cysteinate and one or two oxygenic ligands. Iron-bound (Nif)IscA undergoes reversible redox cycling between the Fe(III)/Fe(II) forms with a midpoint potential of +36 ± 15 mV at pH 7.8 (versus NHE). l-Cysteine is effective in mediating release of free Fe(II) from both the Fe(II)- and Fe(III)-bound forms of (Nif)IscA. Fe(III)-bound (Nif)IscA was also shown to be a competent iron source for in vitro NifS-mediated [2Fe-2S] cluster assembly on the N-terminal domain of NifU, but the reaction occurs via cysteine-mediated release of free Fe(II) rather than direct iron transfer. The proposed roles of A-type proteins in storing Fe under aerobic growth conditions and serving as iron donors for cluster assembly on U-type scaffold proteins or maturation of biological [4Fe-4S] centers are discussed in light of these results.

  9. The Borexino purification system (United States)

    Benziger, Jay


    Purification of 278 tons of liquid scintillator and 889 tons of buffer shielding for the Borexino solar neutrino detector is performed with a system of combined distillation, water extraction, gas stripping and filtration. The purification system removed K, U and Th by distillation of the pseudocumene solvent and the PPO fluor. Noble gases, Rn, Kr and Ar were removed by gas stripping. Distillation was also employed to remove optical impurities and reduce the attenuation of scintillation light. The success of the purification system has facilitated the first time real time detection of low energy solar neutrinos.

  10. Site-directed mutagenesis of Azotobacter vinelandii ferredoxin I: [Fe-S] cluster-driven protein rearrangement

    International Nuclear Information System (INIS)

    Martin, A.E.; Burgess, B.K.; Stout, C.D.; Cash, V.L.; Dean, D.R.; Jensen, G.M.; Stephens, P.J.


    Azotobacter vinelandii ferredoxin I is a small protein that contains one [4Fe-4S] cluster and one [3Fe-4S] cluster. Recently the x-ray crystal structure has been redetermined and the fdxA gene, which encodes the protein, has been cloned and sequenced. Here the authors report the site-directed mutation of Cys-20, which is a ligand of the [4Fe-4S] cluster in the native protein, to alanine and the characterization of the protein product by x-ray crystallographic and spectroscopic methods. The data show that the mutant protein again contains one [4Fe-4S] cluster and one [3Fe-4S] cluster. The new [4Fe-4S] cluster obtains its fourth ligand from Cys-24, a free cysteine in the native structure. The formation of this [4Fe-4S] cluster drives rearrangement of the protein structure

  11. Gel purification of RNA. (United States)

    Nilsen, Timothy W


    For many applications, including size selection of RNAs and purification of in vitro transcription products, it is necessary to purify RNAs on a denaturing gel. This procedure describes how to purify transcripts that have been synthesized in vitro. It is useful for labeled or unlabeled RNAs when sufficient mass is present. It can also be used to isolate small RNAs. In general, RNA purification by denaturing gel electrophoresis is practical only when the size of the desired RNA is 600 nucleotides or less.

  12. Solid State Air Purification System (United States)

    National Aeronautics and Space Administration — The solid state air purification project will explore feasibility of a new air purification system based on a liquid membrane, capable of purifying carbon dioxide...

  13. Purification of lipases. (United States)

    Taipa, M A; Aires-Barros, M R; Cabral, J M


    Interest on lipases from different sources (microorganisms, animals and plants) has markedly increased in the last decade due to the potential applications of lipases in industry and in medicine. Microbial and mammalian lipases have been purified to homogeneity, allowing the successful determination of their primary aminoacid sequence and, more recently, of the three-dimensional structure. The X-ray studies of pure lipases will enable the establishment of the structure-function relationships and contribute for a better understanding of the kinetic mechanisms of lipase action on hydrolysis, synthesis and group exchange of esters. This article reviews the separation and purification techniques that were used in the recovery of microbial, mammalian and plant lipases. Several purification procedures are analysed taking into account the sequence of the methods and the number of times each method is used. Novel purification methods based on liquid-liquid extraction, membrane processes and immunopurification are also reviewed.

  14. Water purification in Borexino

    Energy Technology Data Exchange (ETDEWEB)

    Giammarchi, M. [Infn Milano (Italy); Balata, M.; Ioannucci, L.; Nisi, S. [Laboratori Nazionali del Gran Sasso (Italy); Goretti, A.; Ianni, A. [Princeton University (United States); Miramonti, L. [Dip. di Fisica dell' Università di Milano e Infn (Italy)


    Astroparticle Physics and Underground experiments searching for rare nuclear events, need high purity materials to act as detectors or detector shielding. Water has the advantage of being cheap, dense and easily available. Most of all, water can be purified to the goal of obatining a high level of radiopurity. Water Purification can be achieved by means of a combination of processes, including filtration, reverse osmosis, deionization and gas stripping. The Water Purification System for the Borexino experiment, will be described together with its main performances.

  15. Sodium purification in Rapsodie

    International Nuclear Information System (INIS)

    Giraud, B.


    This report is one of a series of publications presenting the main results of tests carried out during the start-up of the first french fast neutron reactor: Rapsodie. The article presents the sodium purification techniques used in the reactor cooling circuits both from the constructional point of view and with respect to results obtained during the first years working. (author) [fr

  16. Uranium hexafluoride purification

    International Nuclear Information System (INIS)

    Araujo, Eneas F. de


    Uranium hexafluoride might contain a large amount of impurities after manufacturing or handling. Three usual methods of purification of uranium hexafluoride were presented: selective sorption, sublimation, and distillation. Since uranium hexafluoride usually is contaminated with hydrogen fluoride, a theoretical study of the phase equilibrium properties was performed for the binary system UF 6 -HF. A large deviation from the ideal solution behaviour was observed. A purification unity based on a constant reflux batch distillation process was developed. A procedure was established in order to design the re boiler, condenser and packed columns for the UF 6 -HF mixture separation. A bench scale facility for fractional distillation of uranium hexafluoride was described. Basic operations for that facility and results extracted from several batches were discussed. (author)

  17. Mutant Forms of the Azotobacter vinelandii Transcriptional Activator NifA Resistant to Inhibition by the NifL Regulatory Protein


    Reyes-Ramirez, Francisca; Little, Richard; Dixon, Ray


    The Azotobacter vinelandii σ54-dependent transcriptional activator protein NifA is regulated by the NifL protein in response to redox, carbon, and nitrogen status. Under conditions inappropriate for nitrogen fixation, NifL inhibits transcription activation by NifA through the formation of the NifL-NifA protein complex. NifL inhibits the ATPase activity of the central AAA+ domain of NifA required to drive open complex formation by σ54-RNA polymerase and may also inhibit the activator-polymeras...

  18. Modeling and Re-Engineering of Azotobacter vinelandii Alginate Lyase to Enhance Its Catalytic Efficiency for Accelerating Biofilm Degradation.

    Directory of Open Access Journals (Sweden)

    Chul Ho Jang

    Full Text Available Alginate is known to prevent elimination of Pseudomonas aeruginosa biofilms. Alginate lyase (AlgL might therefore facilitate treatment of Pseudomonas aeruginosa-infected cystic fibrosis patients. However, the catalytic activity of wild-type AlgL is not sufficiently high. Therefore, molecular modeling and site-directed mutagenesis of AlgL might assist in enzyme engineering for therapeutic development. AlgL, isolated from Azotobacter vinelandii, catalyzes depolymerization of alginate via a β-elimination reaction. AlgL was modeled based on the crystal structure template of Sphingomonas AlgL species A1-III. Based on this computational analysis, AlgL was subjected to site-directed mutagenesis to improve its catalytic activity. The kcat/Km of the K194E mutant showed a nearly 5-fold increase against the acetylated alginate substrate, as compared to the wild-type. Double and triple mutants (K194E/K245D, K245D/K319A, K194E/K245D/E312D, and K194E/K245D/K319A were also prepared. The most potent mutant was observed to be K194E/K245D/K319A, which has a 10-fold improved kcat value (against acetylated alginate compared to the wild-type enzyme. The antibiofilm effect of both AlgL forms was identified in combination with piperacillin/tazobactam (PT and the disruption effect was significantly higher in mutant AlgL combined with PT than wild-type AlgL. However, for both the wild-type and K194E/K245D/K319A mutant, the use of the AlgL enzyme alone did not show significant antibiofilm effect.

  19. Rapid Biosynthesis of AgNPs Using Soil Bacterium Azotobacter vinelandii With Promising Antioxidant and Antibacterial Activities for Biomedical Applications (United States)

    Karunakaran, Gopalu; Jagathambal, Matheswaran; Gusev, Alexander; Torres, Juan Antonio Lopez; Kolesnikov, Evgeny; Kuznetsov, Denis


    Silver nanoparticles (AgNPs) are applied in various fields from electronics to biomedical applications as a result of their high surface-to-volume ratio. Even though different approaches are available for synthesis of AgNPs, a nontoxic method for the synthesis has not yet been developed. Thus, this study focused on developing an easy and ecofriendly approach to synthesize AgNPs using Azotobacter vinelandii culture extracts. The biosynthesized nanoparticles were further characterized by ultraviolet-visible (UV-Vis) spectroscopy, x-ray diffraction (XRD), Fourier transform infrared (FTIR), energy-dispersive spectrum, particle size distribution (PSD), and transmission electron microscopy (TEM). UV absorption noticed at 435 nm showed formation of AgNPs. The XRD pattern showed a face-centered cubic structure with broad peaks of 28.2°, 32.6°, 46.6°, 55.2°, 57.9°, and 67.8°. The FTIR confirmed the involvement of various functional groups in the biosynthesis of AgNPs. The PSD and TEM analyses showed spherical, well-distributed nanoparticles with an average size of 20-70 nm. The elemental studies confirmed the existence of pure AgNPs. The bacterial extract containing extracellular enzyme nitrate reductase converted silver nitrate into AgNPs. AgNPs significantly inhibited the growth of pathogenic bacteria such as Streptomyces fradiae (National Collection of Industrial Microorganisms (NCIM) 2419), Staphylococcus aureus (NCIM 2127), Escherichia coli (NCIM 2065), and Serratia marcescens (NCIM 2919). In addition, biosynthesized AgNPs were found to possess strong antioxidant activity. Thus, the results of this study revealed that biosynthesized AgNPs could serve as a lead in the development of nanomedicine.

  20. The Effect of Inoculation with Azotobacter and Nitrogen Levels on Grain and Corn Yield Components at Simultaneous Cropping System with Legumes

    Directory of Open Access Journals (Sweden)

    mohammad mirzakhani


    Full Text Available Introduction: Corn has been regarded as one of the important crops from the view point of both human and animal feeding resource. Intercropping defined as cultivation of two or more species together. The advantages of intercropping can be included: efficient use of water and sunlight, exchange of nutrients, weed competition reduction, reduction of pathogens and the increase of soil fertility. Research shows that intercropping combinations of legume–grass will increase forage quality. Because, grasses Grains have a lot of carbohydrates and legumes are rich in protein and vitamins. This study was conducted to evaluate the effect of inoculation with azotobacter and nitrogen levels on grain and corn yield components at simultaneous cropping system with legumes under the weather conditions of Markazi province. Materials and methods: This study was carried out at agricultural research field of Payame Noor University, Arak Branch during 2011. A factorial arrangement of treatment in a randomized complete block design with three replications was used. Methods of plant nutrition (M0= inoculation with azotobacter, M1= inoculation with azotobacter + 37/5 Kg ha-1 of rare nitrogen with foliar application method, M2= inoculation with azotobacter + 150 Kg ha-1 of rare nitrogen mix with soil and simultaneous cropping treatment of legumes, [S1= corn + alfalfa (Medicago sativa L., S2= corn + bitter vetch (Lathyrus sativus L., S3= corn + mung bean (Vigna radiata L., S4= corn + chickpea (Cicer arientinum L., S5= corn + vetch (Vicia ervillia L. ] were assigned in plots. Each sub plot consisted of 4 rows, 6 m long with 60 cm between rows space and 20 cm between plants on the rows and S.C Apex hybrid was used. In this study characteristics such as: plant height, earing height, the number of grains per m-2, the number of rows per ear, the number of grains per row, surface of ear leaf, grain yield of corn, 1000 grain weight, harvest index of corn, nitrogen use

  1. The purification of hypertensin II. (United States)



    The enzymatic conversion of hypertensin I to hypertensin II is described together with the subsequent purification of the product by means of counter-current distribution. Improved methods are also presented for the preparation of renin and its substrate, as well as in methods for the reaction of these materials and the purification of the resulting hypertensin I.

  2. Protein production and purification. (United States)

    Gräslund, Susanne; Nordlund, Pär; Weigelt, Johan; Hallberg, B Martin; Bray, James; Gileadi, Opher; Knapp, Stefan; Oppermann, Udo; Arrowsmith, Cheryl; Hui, Raymond; Ming, Jinrong; dhe-Paganon, Sirano; Park, Hee-won; Savchenko, Alexei; Yee, Adelinda; Edwards, Aled; Vincentelli, Renaud; Cambillau, Christian; Kim, Rosalind; Kim, Sung-Hou; Rao, Zihe; Shi, Yunyu; Terwilliger, Thomas C; Kim, Chang-Yub; Hung, Li-Wei; Waldo, Geoffrey S; Peleg, Yoav; Albeck, Shira; Unger, Tamar; Dym, Orly; Prilusky, Jaime; Sussman, Joel L; Stevens, Ray C; Lesley, Scott A; Wilson, Ian A; Joachimiak, Andrzej; Collart, Frank; Dementieva, Irina; Donnelly, Mark I; Eschenfeldt, William H; Kim, Youngchang; Stols, Lucy; Wu, Ruying; Zhou, Min; Burley, Stephen K; Emtage, J Spencer; Sauder, J Michael; Thompson, Devon; Bain, Kevin; Luz, John; Gheyi, Tarun; Zhang, Fred; Atwell, Shane; Almo, Steven C; Bonanno, Jeffrey B; Fiser, Andras; Swaminathan, Sivasubramanian; Studier, F William; Chance, Mark R; Sali, Andrej; Acton, Thomas B; Xiao, Rong; Zhao, Li; Ma, Li Chung; Hunt, John F; Tong, Liang; Cunningham, Kellie; Inouye, Masayori; Anderson, Stephen; Janjua, Heleema; Shastry, Ritu; Ho, Chi Kent; Wang, Dongyan; Wang, Huang; Jiang, Mei; Montelione, Gaetano T; Stuart, David I; Owens, Raymond J; Daenke, Susan; Schütz, Anja; Heinemann, Udo; Yokoyama, Shigeyuki; Büssow, Konrad; Gunsalus, Kristin C


    In selecting a method to produce a recombinant protein, a researcher is faced with a bewildering array of choices as to where to start. To facilitate decision-making, we describe a consensus 'what to try first' strategy based on our collective analysis of the expression and purification of over 10,000 different proteins. This review presents methods that could be applied at the outset of any project, a prioritized list of alternate strategies and a list of pitfalls that trip many new investigators.

  3. Gas purification project

    International Nuclear Information System (INIS)

    Broothaerts, J.; Claes, J.; Collard, G.; Goossens, W.; Harnie, R.; Heylen, P.; Vaesen, J.; Beukelaer, R. de; Dubois, G.; Glibert, R.; Mestrez, J.; Zahlen, A.


    Conceptual and experimental studies on LMFBR reprocessing and reactor off-gas purification systems are summarized. Iodine sorption on zeolites, low-temperature adsorption of noble gases on charcoal and catalytic oxidation of hydrogen, simulating tritium, are being studied in laboratory set-ups. A pilot loop with 25 m 3 h -1 throughput has been constructed. Results are quoted from the first phase of the iodine removal programme by scrubbing systems. Further extension of the test loop, comprising off-gases conditioning to removal of krypton in a cryodistillation unit, has been prepared. Delay-bed studies on 133 Xe extraction from LWR off-gases are reported. (author)

  4. Air/Water Purification (United States)


    After 18 years of research into air/water pollution at Stennis Space Center, Dr. B. C. Wolverton formed his own company, Wolverton Environmental Services, Inc., to provide technology and consultation in air and water treatment. Common houseplants are used to absorb potentially harmful materials from bathrooms and kitchens. The plants are fertilized, air is purified, and wastewater is converted to clean water. More than 100 U.S. communities have adopted Wolverton's earlier water hyacinth and artificial marsh applications. Catfish farmers are currently evaluating the artificial marsh technology as a purification system.

  5. Water Purification Product (United States)


    Ecomaster, an affiliate of BioServe Space Technologies, this PentaPure technology has been used to purify water for our nation's Space Shuttle missions since 1981. WTC-Ecomaster of Mirneapolis, Minnesota manufactures water purification systems under the brand name PentaPure (TM). BioServe researcher Dr. George Marchin, of Kansas State University, first demonstrated the superiority of this technology and licensed it to WTC. Marchin continues to perform microgravity research in the development of new technologies for the benefit of life on Earth.

  6. Spectroscopic and functional characterization of iron-sulfur cluster-bound forms of Azotobacter vinelandii (Nif)IscA. (United States)

    Mapolelo, Daphne T; Zhang, Bo; Naik, Sunil G; Huynh, Boi Hanh; Johnson, Michael K


    The mechanism of [4Fe-4S] cluster assembly on A-type Fe-S cluster assembly proteins, in general, and the specific role of (Nif)IscA in the maturation of nitrogen fixation proteins are currently unknown. To address these questions, in vitro spectroscopic studies (UV-visible absorption/CD, resonance Raman and Mössbauer) have been used to investigate the mechanism of [4Fe-4S] cluster assembly on Azotobacter vinelandii(Nif)IscA, and the ability of (Nif)IscA to accept clusters from NifU and to donate clusters to the apo form of the nitrogenase Fe-protein. The results show that (Nif)IscA can rapidly and reversibly cycle between forms containing one [2Fe-2S](2+) and one [4Fe-4S](2+) cluster per homodimer via DTT-induced two-electron reductive coupling of two [2Fe-2S](2+) clusters and O(2)-induced [4Fe-4S](2+) oxidative cleavage. This unique type of cluster interconversion in response to cellular redox status and oxygen levels is likely to be important for the specific role of A-type proteins in the maturation of [4Fe-4S] cluster-containing proteins under aerobic growth or oxidative stress conditions. Only the [4Fe-4S](2+)-(Nif)IscA was competent for rapid activation of apo-nitrogenase Fe protein under anaerobic conditions. Apo-(Nif)IscA was shown to accept clusters from [4Fe-4S] cluster-bound NifU via rapid intact cluster transfer, indicating a potential role as a cluster carrier for delivery of clusters assembled on NifU. Overall the results support the proposal that A-type proteins can function as carrier proteins for clusters assembled on U-type proteins and suggest that they are likely to supply [2Fe-2S] clusters rather than [4Fe-4S] for the maturation of [4Fe-4S] cluster-containing proteins under aerobic or oxidative stress growth conditions.

  7. Effect of charcoal on water purification


    Suzuki, Hirotaka; Kawahigashi, Tatsuo


    [Abstract] A natural basin system purifies water through self-purification, but the water pollution load of a river might exceed its self-purification capacity. Charcoal, which is used for other uses aside from heating, such as air purification, was evaluated experimentally for water quality purification. The experiment described herein is based on simple water quality measurements. Some experimentally obtained results are discussed.

  8. Purification technologies for colloidal nanocrystals. (United States)

    Shen, Yi; Gee, Megan Y; Greytak, A B


    Almost all applications of colloidal nanocrystals require some type of purification or surface modification process following nanocrystal growth. Nanocrystal purification - the separation of nanocrystals from undesired solution components - can perturb the surface chemistry and thereby the physical properties of colloidal nanocrystals due to changes in solvent, solute concentrations, and exposure of the nanocrystal surface to oxidation or hydrolysis. For example, nanocrystal quantum dots frequently exhibit decreased photoluminescence brightness after precipitation from the growth solvent and subsequent redissolution. Consequently, purification is an integral part of the synthetic chemistry of colloidal nanocrystals, and the effect of purification methods must be considered in order to accurately compare and predict the behavior of otherwise similar nanocrystal samples. In this Feature Article we examine established and emerging approaches to the purification of colloidal nanoparticles from a nanocrystal surface chemistry viewpoint. Purification is generally achieved by exploiting differences in properties between the impurities and the nanoparticles. Three distinct properties are typically manipulated: polarity (relative solubility), electrophoretic mobility, and size. We discuss precipitation, extraction, electrophoretic methods, and size-based methods including ultracentrifugation, ultrafiltration, diafiltration, and size-exclusion chromatography. The susceptibility of quantum dots to changes in surface chemistry, with changes in photoluminescence decay associated with surface chemical changes, extends even into the case of core/shell structures. Accordingly, the goal of a more complete description of quantum dot surface chemistry has been a driver of innovation in colloidal nanocrystal purification methods. We specifically examine the effect of purification on surface chemistry and photoluminescence in quantum dots as an example of the challenges associated with

  9. Magnetic Purification of Antibodies (United States)

    Dhadge, Vijaykumar Laxman

    This work aimed at the development of magnetic nanoparticles for antibody purification and at the evaluation of their performance in Magnetic fishing and in a newly developed hybrid technology Magnetic Aqueous Two Phase Systems. Magnetic materials were produced by coprecipitation and solvothermal approaches. Natural polymers such as dextran, extracellular polysaccharide and gum Arabic were employed for coating of iron oxide magnetic supports. Polymer coated magnetic supports were then modified with synthetic antibody specific ligands,namely boronic acid, a triazine ligand (named 22/8) and an Ugi ligand (named A2C7I1). To optimize the efficacy of magnetic nanoparticles for antibody magnetic fishing, various solutions of pure and crude antibody solutions along with BSA as a non-specific binding protein were tested. The selectivity of magnetic nanoparticle for antibody, IgG, was found effective with boronic acid and ligand 22/8. Magnetic supports were then studied for their performance in high gradient magnetic separator for effective separation capability as well as higher volume handling capability. The magnetic materials were also supplemented to aqueous two phase systems, devising a new purification technology. For this purpose, magnetic particles modified with boronic acid were more effective. This alternative strategy reduced the time of operation,maximized separation capability (yield and purity), while reducing the amount of salt required. Boronic acid coated magnetic particles bound 170 +/- 10 mg hIgG/g MP and eluted 160 +/- 5 mg hIgG/g MP, while binding only 15 +/- 5 mg BSA/g MP. The affinity constant for the interaction between hIgG and APBA_MP was estimated as 4.9 x 105 M-1 (Ka) with a theoretical maximum capacity of 492 mg hIgG adsorbed/g MP (Qmax). APBA_MPs were also tested for antibody purification directly from CHO cell supernatants. The particles were able to bind 98% of IgG loaded and to recover 95% of pure IgG (purity greater than 98%) at extremely

  10. Effect of Nitrogen and Zinc Sulphate Fertilizers and Azotobacter and Azospirillum Biofertilizer on Yield and Growth Traits of Rapeseed (Brassica napus L.

    Directory of Open Access Journals (Sweden)

    N. Jafari


    Full Text Available In order to study the effects of simultaneous application of nitrogen (N and ZnSO4 fertilizers and biofertilizer (Azotobacter and Azospirillum on grain yield and growth traits of rapeseed, Hyola308 cultivar, a field experiment, with split plot factorial layout based on randomized complete blocks design with three replications, was conducted at Research Field of Faculty of Agriculture, University of Guilan, Rasht, Iran, during 2007-2008 growing season. Nitrogen fertilizer at four levels (0, 50, 100 and 150 kg/ha were the main plot and ZnSO4 fertilizer at two levels (0 and 50 kg/ha and biofertilizer at two levels (with and without biofertilizer were arranged in sub-plots. Results showed that maximum and minimum leaf area indices at flowering stage (average of 1.29 and 0.95, respectively were obtained in 150 kg/ha N+ZnSO4+ biofertilizer and in 50 kg/ha N+ no ZnSO4+ no biofertilizer treatments. Maximum and minimum crop growth rates at flowering stage (average of 5.89 and 3.19 g/m2.GDD, respectively were obtained in 150 kg/ha N+ZnSO4+ biofertilizer and control treatments. Maximum and minimum grain yields (2568, 2468 and 543 kg/ha, respectively were obtained in 150 kg/ha N+ with/without ZnSO4+ biofertilizer and control (no fertilizer treatments. Maximum and minimum oil yields (42.8 and 37.3%, respectively were measured in 0 kg/ha N+ZnSO4+ biofertilizer and 150 kg/ha N+ no ZnSO4+ no biofertilizer treatments. Since there was no significant difference between 150 and 100 kg/ha N+ZnSO4+ biofertilizer treatments in terms of impact on canola grain yield and growth traits, it seems that application of biofertilizer (Azotobacter and Azospirillum, without any reduction in yield, increased grain production and oil content and saved 50 kg/ha of N fertilizer. Biofertilizer (Azotobacter and Azospirillum, along with zinc and sulfur, produced phytohormones, and N fertilizer increased dry matter accumulation and leaf area index (by increasing carbohydrate conversion

  11. Acrylic purification and coatings

    International Nuclear Information System (INIS)

    Kuzniak, Marcin


    Radon (Rn) and its decay daughters are a well-known source of background in direct WIMP detection experiments, as either a Rn decay daughter or an alpha particle emitted from a thin inner surface layer of a detector could produce a WIMP-like signal. Different surface treatment and cleaning techniques have been employed in the past to remove this type of contamination. A new method of dealing with the problem has been proposed and used for a prototype acrylic DEAP-1 detector. Inner surfaces of the detector were coated with a layer of ultra pure acrylic, meant to shield the active volume from alphas and recoiling nuclei. An acrylic purification technique and two coating techniques are described: a solvent-borne (tested on DEAP-1) and solvent-less (being developed for the full scale DEAP-3600 detector).

  12. Caracterización de cepas nativas de Azotobacter spp. y su efecto en el desarrollo de Lycopersicon esculentum Mill. “tomate” en Lambayeque

    Directory of Open Access Journals (Sweden)

    Cynthia Escobar


    Full Text Available El objetivo de la presente investigación fue caracterizar y determinar el efecto de cepas nativas de Azotobacter spp. en el desarrollo vegetativo de Lycopersicon esculentum Mill. “tomate”, como una alternativa al uso indiscriminado de fertilizantes químicos. Se tomaron muestras de raíces y suelo rizosférico de hortalizas con las que se realizaron diluciones (10-4 en caldo Ashby-Sacarosa y se incubaron a 30 ºC hasta observar un color amarillo, turbidez y película superficial. El género Azotobacter se identificó en agar mineral sin nitrógeno y Ashby-Benzoato, obteniéndose 96 cepas con una producción de 7.10 a 57.99 mgL-1 de ácido indolacético, 0.13 a 1.64 mgL-1 de nitrógeno fijado como amonio y hasta 1.61 % de eficiencia en la solubilización de roca fosfórica de Bayóvar. Se obtuvo una suspensión celular (108 de cada una de las cuatro cepas con los mayores valores y se inocularon independientemente y en consorcio, así como una combinación con 50 % de urea-100 % de roca fosfórica, en la rizósfera de tomate cv. Río Grande, en un diseño experimental completamente aleatorio. Todas las cepas nativas incrementaron la altura, volumen radicular, materia seca total, parte aérea y radicular frente al testigo absoluto.

  13. Effects of Azotobacter and Azospirillum and Levels of Manure on Quantitative and Qualitative Traits of Safflower (Carthamus tinctorius L.

    Directory of Open Access Journals (Sweden)

    maryam shahraki


    Full Text Available Introduction The demand for food and agricultural products are increasing in a line of population increasing in the world (Alexandratos, 2003. It is possible to increase the quality and quantity of agricultural products via extending the farms and producing more products (Astaaraei and Koocheki, 1995. Environmental problems caused by synthetic fertilizers and the high levels of producing and introducing such chemicals, have been encouraged the researchers to apply bio-fertilizers for increasing the production in a frame of sustainable agriculture (Rajendran and Devarj, 2004. In this study, the economical yield and agronomy index of Safflower (Carthamus tinctorius L. in manure and bio fertilizers treatments was studied. Materials and method This study was conducted in Agricultural Research Institute, University of Zabol during winter season, 2013. Safflower seeds were planted in sandy loam with pH 8.2. The experimental design was factorial in a frame on randomized completely blocks with three replications. The manure as a first factor had three levels, including no treatment (control, 20 and 30 t.ha-1, while second factor was bio-fertilizer treatment with 4 levels, including no treatment (control, Azosprilium (Azo, Azotobacter (Azt and combined treatment of Azo+Azt. The processed manure and bacteria obtained from local farmers were used in this study. Populations of 108 bacteria were prepared and 24 hours before sowing, seeds were soaked in bacteria. After land preparation, experimental plots were (2.5 × 2 m2 created and treated seeds were planted (40 plants.m-2 manually and plantation was watered immediately. In this study plant height, number of heads in bush, number of seeds per head, seed weight per head, seed weight, grain and biological yield, harvest index, leaf chlorophyll, protein and oil percent were studied. Economical yield and agronomy indices of Safflower were calculated at the end of the season and data were analyzed using SAS

  14. Water Purification Systems (United States)


    Clearwater Pool Technologies employs NASA-developed silver/copper ionization to purify turtle and dolphin tanks, cooling towers, spas, water recycling systems, etc. The pool purifier consists of a microcomputer to monitor water conditions, a pair of metallic electrodes, and a rheostat controller. Ions are generated by passing a low voltage current through the electrodes; the silver ions kill the bacteria, and the copper ions kill algae. This technology has found broad application because it offers an alternative to chemical disinfectants. It was originally developed to purify water on Apollo spacecraft. Caribbean Clear has been using NASA's silver ionization technology for water purification for more than a decade. Two new products incorporate advancements of the basic technology. One is the AquaKing, a system designed for areas with no source of acceptable drinking water. Another is the Caribbean Clear Controller, designed for commercial pool and water park applications where sanitizing is combined with feedback control of pH and an oxidizer, chlorine or bromine. The technology was originally developed to purify water on Apollo spacecraft.

  15. Rapid purification of recombinant histones. (United States)

    Klinker, Henrike; Haas, Caroline; Harrer, Nadine; Becker, Peter B; Mueller-Planitz, Felix


    The development of methods to assemble nucleosomes from recombinant histones decades ago has transformed chromatin research. Nevertheless, nucleosome reconstitution remains time consuming to this day, not least because the four individual histones must be purified first. Here, we present a streamlined purification protocol of recombinant histones from bacteria. We termed this method "rapid histone purification" (RHP) as it circumvents isolation of inclusion bodies and thereby cuts out the most time-consuming step of traditional purification protocols. Instead of inclusion body isolation, whole cell extracts are prepared under strongly denaturing conditions that directly solubilize inclusion bodies. By ion exchange chromatography, the histones are purified from the extracts. The protocol has been successfully applied to all four canonical Drosophila and human histones. RHP histones and histones that were purified from isolated inclusion bodies had similar purities. The different purification strategies also did not impact the quality of octamers reconstituted from these histones. We expect that the RHP protocol can be readily applied to the purification of canonical histones from other species as well as the numerous histone variants.


    Directory of Open Access Journals (Sweden)

    Azam Khodashenas Pelko


    Full Text Available The Jainism emphasizes three major teachings about the purification of the soul (jiva, Ahimsa, Aparigrapha and anekantwad. Jainism, The focus of this religion has been purification of the soul by means of right conduct, right faith and right knowledge. The ultimate goal of Hinduism is Moksha or liberation (total freedom. In Hinduism, purification of the soul is a goal that one must work to attain. The Buddhism is the science of pursuing the aim of making the human mind perfect, and of purifying the human soul. The knowledge of purifying of the soul and softening of the hearts is as essential for human. They having the correct motivations means purifying our souls from hypocrisy, caprice, and heedlessness. The primary goal of Taoism may be described as the mystical intuition of the Tao, which is the way, the undivided unity, and the ultimate Reality. According to the Christianity access to truth cannot be conceived without purity of the soul

  17. Purification of Water by Aquatic Plants


    Morimitsu, Katsuhito; Kawahigashi, Tatsuo


    [Abstract] Water quality purification of many water systems including those occurring in rivers depends to a great degree on water quality purification activities of aquatic plants and microbes. This paper presents a discussion of results, based on laboratory experiments, of purification by aquatic plants.

  18. Partial Purification and Characterization of Extracellular Protease ...

    African Journals Online (AJOL)

    Nigerian Journal of Basic and Applied Sciences ... Purification of the enzyme by gel filtration chromatography on Sephadex G75 following ammonium sulphate precipitation gave 2.26 fold increase in purification with specific activity of 46.13 units/mg protein while purification on Sephadex CM50 resulted in reduced ...

  19. Determinación del potencial promotor del crecimiento vegetal de Azotobacter spp. aislados de la rizósfera de malezas en cultivos de maíz (Zea mays L.

    Directory of Open Access Journals (Sweden)

    Luis H. León


    Full Text Available Los fertilizantes químicos representan entre 20% y 30% de los costos de producción de un cultivo, utilizados correctamente incrementan la productividad y rentabilidad; sin embargo, cada año aumenta la cantidad de fertilizantes por aplicar, debido a la deficiencia de adsorción en el suelo y absorción por la planta. Siendo el maíz el tercer cultivo de importancia en Perú, con un impacto significativo en la actividad económica y social, en el 2014, solo el 40% del maíz ofertado correspondió a la producción nacional. En busca de alternativas para disminuir el uso de fertilizantes químicos se realizan investigaciones con denominadas rizobacterias promotoras del crecimiento vegetal (PGPR, por sus siglas en ingles. Se identificaron 37 malezas en cultivos de maíz procedentes de campos agrícolas de los distritos de Monsefú y Reque, Región Lambayeque, siendo dicotiledóneas predominantes con 68 % con respecto a monocotiledóneas con 32%. Las bacterias se aislaron de la rizósfera de malezas, obteniendo 305 cultivos puros de bacterias, de los cuales 133 cultivos puros (43,7% se identificaron como Azotobacter spp., investigándose su reacción bioquímica en reducción de nitratos, utilización de sacarosa, glucosa, maltosa, manitol, ramnosa, glicerol y sorbitol, identificándose A. vinelandii (58%, A. paspali (13%, A. armeniacus (8%, A. nigricans (8% y en 13 cultivos no se identificó la especie. Con los cultivos Azotobacter spp. se cuantificó hasta 36,03 ppm de nitrógeno fijado como amonio; 60,75 ppm de ácido indol acético y 6,06 ppm de fósforo solubilizado, se determinó actividad antagónica contra Fusarium verticillioides, proteolítica y quitinolítica y. El 20% de Azotobacter spp. no afectó la emergencia de maíz amarillo duro hibrido simple AGRI- 144, el 33 % la afectó positivamente y el 47% la afectó negativamente. A su vez, ningún Azotobacter spp. afectó la sobrevivencia. Demostrándose el potencial promotor del


    African Journals Online (AJOL)

    Prof. Adipala Ekwamu

    phosphatase activity upon phosphate deprivation with much of the enzyme released into the medium during osmotic shock, indicating .... shaking. Protein purification. Three millilitres of overnight culture (400 ml) was harvested by centrifugation at 6000x g for 1 min. The supernatant was decanted and pellet washed twice in ...

  1. Bioinspired Materials for Water Purification

    Directory of Open Access Journals (Sweden)

    Alfredo Gonzalez-Perez


    Full Text Available Water scarcity issues associated with inadequate access to clean water and sanitation is a ubiquitous problem occurring globally. Addressing future challenges will require a combination of new technological development in water purification and environmental remediation technology with suitable conservation policies. In this scenario, new bioinspired materials will play a pivotal role in the development of more efficient and environmentally friendly solutions. The role of amphiphilic self-assembly on the fabrication of new biomimetic membranes for membrane separation like reverse osmosis is emphasized. Mesoporous support materials for semiconductor growth in the photocatalytic degradation of pollutants and new carriers for immobilization of bacteria in bioreactors are used in the removal and processing of different kind of water pollutants like heavy metals. Obstacles to improve and optimize the fabrication as well as a better understanding of their performance in small-scale and pilot purification systems need to be addressed. However, it is expected that these new biomimetic materials will find their way into the current water purification technologies to improve their purification/removal performance in a cost-effective and environmentally friendly way.

  2. [Isolation and study of azobenzene converting soil bacteria]. (United States)

    Vakkerov-Kouzova, N D


    Heterotrophic bacteria were isolated from soil and glass slides and classified as Bacillus cereus SNK12, Paenibacillus polymyxa SNK2, Azotobacter chroococcum ANKII, and Ochrobacterium intermedium ANKI. Their cultures could degrade azobenzene under the conditions of co-metabolism. A rapid test for the ability of bacteria to convert azobenzenes is proposed.

  3. Characterization of the Paenibacillus beijingensis DSM 24997 GtfD and its glucan polymer products representing a new glycoside hydrolase 70 subfamily of 4,6-α-glucanotransferase enzymes

    NARCIS (Netherlands)

    Gangoiti, Joana; Lamothe, Lisa; van Leeuwen, Sander Sebastiaan; Vafiadi, Christina; Dijkhuizen, Lubbert


    Previously we have reported that the Gram-negative bacterium Azotobacter chroococcum NCIMB 8003 uses the 4,6-α-glucanotransferase GtfD to convert maltodextrins and starch into a reuteran-like polymer consisting of (α1→4) glucan chains connected by alternating (α1→4)/(α1→6) linkages and (α1→4,6)

  4. Plant growth promotion rhizobacteria in onion production. (United States)

    Colo, Josip; Hajnal-Jafari, Timea I; Durić, Simonida; Stamenov, Dragana; Hamidović, Saud


    The aim of the research was to examine the effect of rhizospheric bacteria Azotobacter chroococcum, Pseudomonas fluorescens (strains 1 and 2) and Bacillus subtilis on the growth and yield of onion and on the microorganisms in the rhizosphere of onion. The ability of microorganisms to produce indole-acetic acid (IAA), siderophores and to solubilize tricalcium phosphate (TCP) was also assessed. The experiment was conducted in field conditions, in chernozem type of soil. Bacillus subtilis was the best producer of IAA, whereas Pseudomonas fluorescens strains were better at producing siderophores and solubilizing phosphates. The longest seedling was observed with the application of Azotobacter chroococcum. The height of the plants sixty days after sowing was greater in all the inoculated variants than in the control. The highest onion yield was observed in Bacillus subtilis and Azotobacter chroococcum variants. The total number of bacteria and the number of Azotobacter chroococcum were larger in all the inoculated variants then in the control. The number of fungi decreased in most of the inoculated variants, whereas the number of actinomycetes decreased or remained the same.

  5. Iron environment in ferritin with large amounts of phosphate, from Azotobacter vinelandii and horse spleen, analyzed using Extended X-ray Absorption fine Structure (EXAFS)

    International Nuclear Information System (INIS)

    Rohrer, J.S.; Islam, Q.T.; Sayes, D.E.; Theil, E.C.; Watt, G.D.


    The iron core of proteins in the ferritin family displays structural variations that includes phosphate content was well as the number and the degree of ordering of the iron atoms. Earlier studies had shown that ferritin iron cores naturally high in phosphate, e.g., Azotobacter vinelandii (AV) ferritin had decreased long-range order. Here, the influence of phosphate on the local structure around iron in ferritin cores is reported, comparing the EXAFS of AV ferritin, reconstituted ferritin and native horse spleen ferritin. In contrast, when the phosphate content was high in AV ferritin and horse spleen ferritin reconstituted with phosphate, the average iron atom had five to six phosphorus neighbors at 3.17 angstrom. Moreover, the number of detectable iron neighbors was lower when phosphate was high or present during reconstitution and the interatomic distance was longer indicating that some phosphate bridges neighboring iron atoms. However, the decrease in the number of detectable iron-iron neighbors compared to HSF and the higher number of Fe-P interactions relative to Fe-Fe interactions suggest that some phosphate ligands were chain termini, or blocked crystal growth, and/or introduced defects which contributed both to the long-range disorder and to altered redox properties previously observed in AV ferritin

  6. Ligand size is a major determinant of specificity in periplasmic oxyanion-binding proteins: the 1.2 A resolution crystal structure of Azotobacter vinelandii ModA. (United States)

    Lawson, D M; Williams, C E; Mitchenall, L A; Pau, R N


    . Periplasmic receptors constitute a diverse class of binding proteins that differ widely in size, sequence and ligand specificity. Nevertheless, almost all of them display a common beta/alpha folding motif and have similar tertiary structures consisting of two globular domains. The ligand is bound at the bottom of a deep cleft, which lies at the interface between these two domains. The oxyanion-binding proteins are notable in that they can discriminate between very similar ligands. . Azotobacter vinelandii is unusual in that it possesses two periplasmic molybdate-binding proteins. The crystal structure of one of these with bound ligand has been determined at 1.2 A resolution. It superficially resembles the structure of sulphate-binding protein (SBP) from Salmonella typhimurium and uses a similar constellation of hydrogen-bonding interactions to bind its ligand. However, the detailed interactions are distinct from those of SBP and the more closely related molybdate-binding protein of Escherichia coli. . Despite differences in the residues involved in binding, the volumes of the binding pockets in the A. vinelandii and E. coli molybdate-binding proteins are similar and are significantly larger than that of SBP. We conclude that the discrimination between molybdate and sulphate shown by these binding proteins is largely dependent upon small differences in the sizes of these two oxyanions.

  7. Statistical and Judgmental Criteria for Scale Purification

    DEFF Research Database (Denmark)

    Wieland, Andreas; Durach, Christian F.; Kembro, Joakim


    of scale purification, to critically analyze the current state of scale purification in supply chain management (SCM) research and to provide suggestions for advancing the scale-purification process. Design/methodology/approach A framework for making scale-purification decisions is developed and used......Purpose “Scale purification” – the process of eliminating items from multi-item scales – is widespread in empirical research, but studies that critically examine the implications of this process are scarce. The goals of this research are threefold: to discuss the methodological underpinning...... to analyze and critically reflect on the application of scale purification in leading SCM journals. Findings This research highlights the need for rigorous scale-purification decisions based on both statistical and judgmental criteria. By applying the proposed framework to the SCM discipline, a lack...

  8. Water purification using organic salts (United States)

    Currier, Robert P.


    Water purification using organic salts. Feed water is mixed with at least one organic salt at a temperature sufficiently low to form organic salt hydrate crystals and brine. The crystals are separated from the brine, rinsed, and melted to form an aqueous solution of organic salt. Some of the water is removed from the aqueous organic salt solution. The purified water is collected, and the remaining more concentrated aqueous organic salt solution is reused.

  9. Technological assumptions for biogas purification. (United States)

    Makareviciene, Violeta; Sendzikiene, Egle


    Biogas can be used in the engines of transport vehicles and blended into natural gas networks, but it also requires the removal of carbon dioxide, hydrogen sulphide, and moisture. Biogas purification process flow diagrams have been developed for a process enabling the use of a dolomite suspension, as well as for solutions obtained by the filtration of the suspension, to obtain biogas free of hydrogen sulphide and with a carbon dioxide content that does not exceed 2%. The cost of biogas purification was evaluated on the basis of data on biogas production capacity and biogas production cost obtained from local water treatment facilities. It has been found that, with the use of dolomite suspension, the cost of biogas purification is approximately six times lower than that in the case of using a chemical sorbent such as monoethanolamine. The results showed travelling costs using biogas purified by dolomite suspension are nearly 1.5 time lower than travelling costs using gasoline and slightly lower than travelling costs using mineral diesel fuel.

  10. Comparing Russian and Finnish standards of water purification


    Maria, Pupkova


    The subject of this thesis is water purification. The first aim of this thesis is to consider different ways of water purification. The second aim is to compare Finnish and Russian standards of water purification. The third one is to show water purification methods on the pattern of Mikkeli water purification plan. Water purification methods of water intended for human consumption will be described.Combined tables will be done according to the quality requirement of drinking water of both,...

  11. Inactivation of an intracellular poly-3-hydroxybutyrate depolymerase of Azotobacter vinelandii allows to obtain a polymer of uniform high molecular mass. (United States)

    Adaya, Libertad; Millán, Modesto; Peña, Carlos; Jendrossek, Dieter; Espín, Guadalupe; Tinoco-Valencia, Raunel; Guzmán, Josefina; Pfeiffer, Daniel; Segura, Daniel


    A novel poly-3-hydroxybutyrate depolymerase was identified in Azotobacter vinelandii. This enzyme, now designated PhbZ1, is associated to the poly-3-hydroxybutyrate (PHB) granules and when expressed in Escherichia coli, it showed in vitro PHB depolymerizing activity on native or artificial PHB granules, but not on crystalline PHB. Native PHB (nPHB) granules isolated from a PhbZ1 mutant had a diminished endogenous in vitro hydrolysis of the polyester, when compared to the granules of the wild-type strain. This in vitro degradation was also tested in the presence of free coenzyme A. Thiolytic degradation of the polymer was observed in the nPHB granules of the wild type, resulting in the formation of 3-hydroxybutyryl-CoA, but was absent in the granules of the mutant. It was previously reported that cultures of A. vinelandii OP grown in a bioreactor showed a decrease in the weight average molecular weight (Mw) of the PHB after 20 h of culture, with an increase in the fraction of polymers of lower molecular weight. This decrease was correlated with an increase in the PHB depolymerase activity during the culture. Here, we show that in the phbZ1 mutant, neither the decrease in the Mw nor the appearance of a low molecular weight polymers occurred. In addition, a higher PHB accumulation was observed in the cultures of the phbZ1 mutant. These results suggest that PhbZ1 has a role in the degradation of PHB in cultures in bioreactors and its inactivation allows the production of a polymer of a uniform high molecular weight.

  12. Structural basis for cyclization specificity of two Azotobacter type III polyketide synthases: a single amino acid substitution reverses their cyclization specificity. (United States)

    Satou, Ryutaro; Miyanaga, Akimasa; Ozawa, Hiroki; Funa, Nobutaka; Katsuyama, Yohei; Miyazono, Ken-ichi; Tanokura, Masaru; Ohnishi, Yasuo; Horinouchi, Sueharu


    Type III polyketide synthases (PKSs) show diverse cyclization specificity. We previously characterized two Azotobacter type III PKSs (ArsB and ArsC) with different cyclization specificity. ArsB and ArsC, which share a high sequence identity (71%), produce alkylresorcinols and alkylpyrones through aldol condensation and lactonization of the same polyketomethylene intermediate, respectively. Here we identified a key amino acid residue for the cyclization specificity of each enzyme by site-directed mutagenesis. Trp-281 of ArsB corresponded to Gly-284 of ArsC in the amino acid sequence alignment. The ArsB W281G mutant synthesized alkylpyrone but not alkylresorcinol. In contrast, the ArsC G284W mutant synthesized alkylresorcinol with a small amount of alkylpyrone. These results indicate that this amino acid residue (Trp-281 of ArsB or Gly-284 of ArsC) should occupy a critical position for the cyclization specificity of each enzyme. We then determined crystal structures of the wild-type and G284W ArsC proteins at resolutions of 1.76 and 1.99 Å, respectively. Comparison of these two ArsC structures indicates that the G284W substitution brings a steric wall to the active site cavity, resulting in a significant reduction of the cavity volume. We postulate that the polyketomethylene intermediate can be folded to a suitable form for aldol condensation only in such a relatively narrow cavity of ArsC G284W (and presumably ArsB). This is the first report on the alteration of cyclization specificity from lactonization to aldol condensation for a type III PKS. The ArsC G284W structure is significant as it is the first reported structure of a microbial resorcinol synthase.

  13. The GacS/A-RsmA Signal Transduction Pathway Controls the Synthesis of Alkylresorcinol Lipids that Replace Membrane Phospholipids during Encystment of Azotobacter vinelandii SW136.

    Directory of Open Access Journals (Sweden)

    Yanet Romero

    Full Text Available Azotobacter vinelandii is a soil bacterium that undergoes a differentiation process that forms cysts resistant to desiccation. During encystment, a family of alkylresorcinols lipids (ARs are synthesized and become part of the membrane and are also components of the outer layer covering the cyst, where they play a structural role. The synthesis of ARs in A. vinelandii has been shown to occur by the activity of enzymes encoded in the arsABCD operon. The expression of this operon is activated by ArpR, a LysR-type transcriptional regulator whose transcription occurs during encystment and is dependent on the alternative sigma factor RpoS. In this study, we show that the two component response regulator GacA, the small RNA RsmZ1 and the translational repressor protein RsmA, implicated in the control of the synthesis of other cysts components (i.e., alginate and poly-ß-hydroxybutyrate, are also controlling alkylresorcinol synthesis. This control affects the expression of arsABCD and is exerted through the regulation of arpR expression. We show that RsmA negatively regulates arpR expression by binding its mRNA, repressing its translation. GacA in turn, positively regulates arpR expression through the activation of transcription of RsmZ1, that binds RsmA, counteracting its repressor activity. This regulatory cascade is independent of RpoS. We also show evidence suggesting that GacA exerts an additional regulation on arsABCD expression through an ArpR independent route.

  14. The production, molecular weight and viscosifying power of alginate produced by Azotobacter vinelandii is affected by the carbon source in submerged cultures

    Directory of Open Access Journals (Sweden)

    Mauricio A. Trujillo-Roldán


    Full Text Available El alginato es un polímero lineal compuesto por ácidos 1,4 manurónico y su epímero, -L- gulurónico y con frecuencia se extrae de algas marinas, como también de bacterias como Azotobacter y Pseudomonas. En este trabajo, se presenta el impacto de diferentes fuentes de carbono convencionales y no convencionales en el crecimiento de A. vinelandii, producción de alginato, su peso molecular promedio (PMP y su capacidad viscosificante. Todos los experimentos se iniciaron con 20 g/L de azúcares totales, donde la más alta concentración de biomasa se obtuvo utilizando suero de leche hidrolizado y desproteinizado (6.67±0.72 g/L, y jugo de caña de azúcar (6.68±0.45 g/L. Sin embargo, la producción máxima de alginato se logró utilizando sacarosa (5.11±0.37 g/L, así como el más alto rendimiento de alginato y productividad específica. Por otra parte, el mayor PMP de alginato se obtuvo con jugo de caña de azúcar (1203±120 kDa. Además, la capacidad viscosificante más alta se obtuvo utilizando suero de leche desproteinizado e hidrolizado (23.8±2.6 cpsL/galg. Esta información sugiere que es posible manipular la productividad y las características moleculares de alginatos como función de la fuente de carbono utilizada. En conjunto con el conocimiento de los efectos de las condiciones ambientales se lograrían altos rendimientos de biopolímeros de alto valor agregado.

  15. Purification of rhamnolipid using colloidal magnetic nanoparticles ...

    African Journals Online (AJOL)

    Phospholipid-coated colloidal magnetic nanoparticles with mean magnetite core size of 9 nm are shown to be effective ion exchange media for the recovery and purification of Rhaminolipid from culture mixtures. These particles have high adsorption capacity for purification (an order of magnitude larger than the best ...

  16. Purification of trioctylphosphine oxide by liquid extracting

    International Nuclear Information System (INIS)

    Meddour, Laaldja


    Many methods of TOPO synthesis are known in the litterature. Neverthless, the purification methods still unknown or quite known. In this work, we have proposed to develop a new method of purification and we have used the extraction properties of TOPO. This method consist to extracting the molybdene with TOPO in acid medium

  17. Purification and characterization of amidase from acrylamide ...

    African Journals Online (AJOL)

    An amidase from a newly isolated acrylamide-degrading bacterium Burkholderia sp. strain DR.Y27 was purified to homogeneity by a combination of anion exchange and gel filtration chromatography. The purification strategy achieved 11.15 of purification fold and a yield of 1.55%. The purified amidase consisted of four ...

  18. Ion exchange purification of scandium (United States)

    Herchenroeder, L.A.; Burkholder, H.R.


    An improvement in purification of scandium through ion exchange chromatography is disclosed in which the oxidation potential of the eluting solution is altered by the addition of potassium chlorate or ammonium chloride so that removal of contaminants is encouraged. The temperature, pH and concentration of the eluent HEDTA are controlled in order to maintain the scandium in the column while minimizing dilution of the scandium band. Recovery of scandium is improved by pumping dilute scandium over the column prior to stripping the scandium and precipitation. This eliminates the HEDTA ion and other monovalent cations contaminating the scandium band. This method maximizes recovery of scandium while maintaining purity. 2 figs.

  19. Method for purification of gases

    Energy Technology Data Exchange (ETDEWEB)

    Hastrup, N.E.


    The present invention is directed to a method for selective purification of gases. The method comprises feeding an exhaust gas containing a solid impurity and a gaseous impurity to a separator. A substantial portion of the solid impurity is separated. Then, the partially-purified gas is fed to a spray dryer. That gas is sprayed with an absorption agent to separate gaseous impurities. Then, the further, partially-purified gas is fed to another separator to separate remaining solid impurity. An exhaust gas which is substantially free of solid and gaseous impurities and fly ash having a desired quality are recovered.


    Skeggs, Leonard T.; Marsh, Walton H.; Kahn, Joseph R.; Shumway, Norman P.


    The purification of hypertensin I has been described. The final product which is four times as powerful a pressor agent as l-arterenol, is obtained with an over-all recovery of 40 per cent. The product consists of a single component in countercurrent distribution, having a nitrogen content of 15.97 per cent and a specific activity of 7050 Goldblatt units per mg. of N or 1125 units per mg. of solid. Acid hydrolysis and paper chromatography indicate in a preliminary fashion that there are about nine amino acids present in the intact polypeptide. PMID:13201713

  1. Purification

    DEFF Research Database (Denmark)

    Andersen, Astrid Oberborbeck


    In Arequipa, Peru’s second largest city, engineers work hard to control water flows and provide different sectors with clean and sufficient water. In 2011, only 10 percent of the totality of water used daily by Arequipa’s then close to 1 million people—in households, tourism, industry, and mining......—was treated before it was returned to the river where it continues its flow downstream towards cultivated fields and, finally, into the Pacific Ocean. It takes specialized knowledge and manifold technologies to manage water and sustain life in Arequipa, and engineers are central actors for making water flow....... Examining the ecology of water management, this article asks to what extent we can talk of a way of knowing and enacting water that is particular to engineers. Through engineering practices, a technical domain emerges as separate from and superior to political and social domains. This production...

  2. Kinetic Studies of Iron Deposition Catalyzed by Recombinant Human Liver Heavy, and Light Ferritins and Azotobacter Vinelandii Bacterioferritin Using O2 and H2O2 as Oxidants (United States)

    Bunker, Jared; Lowry, Thomas; Davis, Garrett; Zhang, Bo; Brosnahan, David; Lindsay, Stuart; Costen, Robert; Choi, Sang; Arosio, Paolo; Watt, Gerald D.


    The discrepancy between predicted and measured H2O2 formation during iron deposition with recombinant heavy human liver ferritin (rHF) was attributed to reaction with the iron protein complex [Biochemistry 40 (2001) 10832-10838]. This proposal was examined by stopped-flow kinetic studies and analysis for H2O2 production using (1) rHF, and Azotobacter vinelandii bacterial ferritin (AvBF), each containing 24 identical subunits with ferroxidase centers; (2) site-altered rHF mutants with functional and dysfunctional ferroxidase centers; and (3) rccombinant human liver light ferritin (rLF), containing 110 ferroxidase center. For rHF, nearly identical pseudo-first-order rate constants of 0.18 per second at pH 7.5 were measured for Fe(2+) oxidation by both O2 and H2O2, but for rLF, the rate with O2 was 200-fold slower than that for H2O2 (k-0.22 per second). A Fe(2+)/O2 stoichiometry near 2.4 was measured for rHF and its site altered forms, suggesting formation of H2O2. Direct measurements revealed no H2O2 free in solution 0.5-10 min after all Fe(2+) was oxidized at pH 6.5 or 7.5. These results are consistent with initial H2O2 formation, which rapidly reacts in a secondary reaction with unidentified solution components. Using measured rate constants for rHF, simulations showed that steady-state H2O2 concentrations peaked at 14 pM at approx. 600 ms and decreased to zero at 10-30 s. rLF did not produce measurable H2O2 but apparently conducted the secondary reaction with H2O2. Fe(2+)/O2 values of 4.0 were measured for AvBF. Stopped-flow measurements with AvBF showed that both H2O2 and O2 react at the same rate (k=0.34 per second), that is faster than the reactions with rHF. Simulations suggest that AvBF reduces O2 directly to H2O without intermediate H2O2 formation.

  3. Reverse osmosis water purification system (United States)

    Ahlstrom, H. G.; Hames, P. S.; Menninger, F. J.


    A reverse osmosis water purification system, which uses a programmable controller (PC) as the control system, was designed and built to maintain the cleanliness and level of water for various systems of a 64-m antenna. The installation operates with other equipment of the antenna at the Goldstone Deep Space Communication Complex. The reverse osmosis system was designed to be fully automatic; with the PC, many complex sequential and timed logic networks were easily implemented and are modified. The PC monitors water levels, pressures, flows, control panel requests, and set points on analog meters; with this information various processes are initiated, monitored, modified, halted, or eliminated as required by the equipment being supplied pure water.

  4. Ash study for biogas purification

    International Nuclear Information System (INIS)

    Juarez V, R. I.


    This work evaluates the ashes generated from the wood and coal combustion process of the thermoelectric plant in Petacalco, Guerrero (Mexico) in order to determine its viability as a filter in the biogas purification process. The ash is constituted by particles of morphology and different chemical properties, so it required a characterization of the same by different analytical techniques: as was scanning electron microscopy and X-ray diffraction, in order to observe the microstructure and determine the elemental chemical composition of the particles. Prior to the analysis, a set of sieves was selected to classify as a function of particle size. Four different types of ashes were evaluated: one generated by the wood combustion (wood ash) and three more of the Petacalco thermoelectric generated by the coal combustion (wet fly ash, dry fly ash and dry bottom ash). (Author)

  5. Purification Protocols for Extracellular Vesicles. (United States)

    Lane, Rebecca E; Korbie, Darren; Trau, Matt; Hill, Michelle M


    This chapter provides a description of some of the standard methods used for the isolation of extracellular vesicles (EVs) from a variety of biological fluids, including cell culture media, urine, plasma and serum. The methods presented include ultracentrifugation, ultrafiltration, proprietary polymer-based reagents, size exclusion chromatography, density gradient separation, and immunoaffinity capture. Ultracentrifugation methods use high speed centrifugation to pellet vesicles, whilst polymer-based reagents are added to the sample to facilitate vesicle precipitation using lower speeds. Ultrafiltration involves the concentration of vesicles from a large volume of biological fluid using a centrifugal filter unit. Size exclusion chromatography and density gradient separation are both designed to allow the separation of vesicles from other nonvesicular debris. Immunoaffinity capture methods use antibody-coated beads to selectively isolate vesicles displaying a surface marker of interest. Ultimately, the choice of purification method for an individual experiment is influenced by time, cost, and equipment considerations, as well as the sample requirements for any downstream analyses.

  6. Nanomechanical Water Purification Device, Phase II (United States)

    National Aeronautics and Space Administration — Seldon Laboratories, LLC, proposes a lightweight, low-pressure water purification device that harnesses the unique properties of carbon nanotubes and will operate...

  7. Purification and characterization of thermostable glucoamylase from ...

    African Journals Online (AJOL)

    Thermostable glucoamylase from Rhizopus oligosporus SK5 mutant was purified in a 3-step purification using Imarsil, activated charcoal and Sephadex-G-100 to achieve a 40-fold purification. The enzyme was optimally active at pH 5.0 and temperature of 80 °C. It exhibited a half-life of 60 minutes at 70 °C. Its stability was ...

  8. A metodologia de superfície de resposta como ferramenta para a avaliação da produção de alginato e poli-hidroxibutirato pela Azotobacter vinelandii = The response surface methodology as a tool for assessing the production of alginate and polyhydroxybutirate by Azotobacter vinelandii

    Directory of Open Access Journals (Sweden)

    Adriana Navarro da Silva


    Full Text Available O alginato é um polissacarídeo normalmente extraído de paredes celulares de algas marrons e utilizado nas indústrias de alimentos, farmacêuticas e biotecnológicas. A produção é concentrada no cultivo de algas marinhas marrons, mas várias bactérias do gênero Pseudomonas e Azotobacter produzem alginato. A estrutura química dos alginatosproduzidos por algas é similar a dos sintetizados pela A. vinelandii. Esta bactéria também produz polímeros intracelulares como o poli-hidroxibutirato (PHB, conhecido como bioplástico. Neste trabalho, estudou-se a produção simultânea do alginato e PHB pela A.vinelandii, utilizando-se sacarose e diferentes parâmetros de fermentação em agitador orbital rotatório. Os valores ótimos para produção destes compostos foram determinados pela MSR. O 1º experimento foi um planejamento fatorial fracionado 26-2. O 2º foi baseado nas variáveis significativas do 1º experimento, resultando em um planejamento fatorial completo 33-0. Verificou-se, do primeiro para o segundo, aumento na produtividade do PHB de 12 para 45 mg g-1 de célula h-1 e do alginato de 100 para 1.600 mg g-1 de célula h-1. Aprodutividade de ambos os compostos foi máxima na temperatura de incubação de 62ºC, no menor tempo de incubação (18h e na concentração de sacarose de 11 g L-1. Em ambos os experimentos, o PHB extraído apresentou pureza de 94%.Alginate is a polysaccharide extracted from cell walls of brown algae and used in the food, pharmaceuticals and biotech industries. Production is concentrated on the cultivation of brown seaweed, but several bacteria of the genus Pseudomonas and Azotobacter producealginate. The chemical structure of alginates produced by algae is similar to those synthesized by A. vinelandii. The bacteria also produce intracellular polymers such as polyhydroxybutyrate (PHB, known as bioplastic. This work studied the simultaneous alginate and PHB production by A. vinelandii using sucrose and

  9. Assessing the Effect of Organic Compounds, Biofertilizers and Chemical Fertilizers on Morphological Properties,yield and Yield Components of Forage Sorghum (Sorghum bicolor

    Directory of Open Access Journals (Sweden)

    A.H Saeidnejad


    Full Text Available Recently, using the source of organic fertilizers and biofertilizers in sustainable crop production is growing. In order to evaluate the effect of organic compounds, biofertilizers and chemical fertilizer on morphological properties, yield and yield components of forage Sorghum (sorghum bicolor a field experiment was conducted in the Research Farm, College of Agriculture, Ferdowsi University of Mashhad in 2008.The treatments were seed inoculation with the combination of Azotobacter chroococcum and Azospirillum brasilense, Compost (15 t/ha, Vermicompost (10 t/ha, seed inoculation with Azotobacter and Azospirillum and compost (10t/ha, seed inoculation with Azotobacter chroococcum and Azospirillum brasilense and Vermicompost (7t/ha, seed inoculation with Pseudomonas flurescence, seed inoculation with Pseudomonas flurescence and Azotobacter chroococcum and Azospirillum brasilense combination, seed inoculation with Pseudomonas flurescence and compost (15t/ha, chemical fertilizer (80 kg/h urea fertilizer and 50 kg/h super phosphate fertilizer and control. Harvesting was performed in 2 cuts in flowering stage. Plant height, number of tiller per plant and SPAD reading was significantly affected by the treatments. Stem diameter was not affected by any treatments. There was a significant difference among all treatments in terms of fresh and dry forage yield. There were no significant differences among all treatments in terms of stem and leaf dry matter. In general, result of this experiment indicated that organic amendments and biofertilizers could be acceptable alternatives for chemical fertilizers.

  10. Purification processes for coal gasification

    Energy Technology Data Exchange (ETDEWEB)

    Fleming, D.K.; Primack, H.S.


    It is apparent from the discussion that many routes can be taken to achieve acid-gas removal and sulfur recovery from coal gas. The selection of the optimum purification system is a major task. The type of coal, type of gasifier and the upstream processing all strongly influence the selection. Several generalizations can be made: (1) The cost of the purification sections of a high-Btu gas plant is significant--perhaps 10 to 30% of the capital cost of the coal conversion facility. (2) The cost of purifying gas produced from high-sulfur coal feed is more expensive than the cost for purifying gas produced from low-sulfur coal. (3) The choice of an acid-gas removal system will often be a function of system pressure. The economical choice will usually be: (a) amine-based systems at atmospheric pressure; (b) hot-carbonate systems at moderate pressure or (c) physical-solvent systems at higher pressure. (4) For a high-Btu, high-sulfur case: (a) A selective acid-gas removal system with a Claus plant is probably more economical than a non-selective acid-gas system with liquid oxidation of the H/sub 2/S in the regenerator off-gas. (b) Even moderately selective systems can produce an H/sub 2/S-rich gas suitable for a Claus plant. The CO/sub 2/-rich gas may or may not require further sulfur removal, depending on the selectivity. (5) For a high-Btu, low-sulfur case: (a) The hot carbonate and tertiary amine systems may not be sufficiently selective to produce a gas suitable for feed to a Claus process while a physical solvent system may be. Therefore, the physical solvent system may be expected to be more economical. (b) The regenerated gas from the bulk CO/sub 2/ removal system following a selective physical solvent system may require further sulfur removal, depending upon the sulfur level in the initial feedstock and the selectivity of the system selected.

  11. Dialysis membranes for blood purification. (United States)

    Sakai, K


    All of the artificial membranes in industrial use, such as a reverse-osmosis membrane, dialysis membrane, ultrafiltration membrane, microfiltration membrane and gas separation membrane, also have therapeutic applications. The most commonly used artificial organ is the artificial kidney, a machine that performs treatment known as hemodialysis. This process cleanses the body of a patient with renal failure by dialysis and filtration, simple physicochemical processes. Hemodialysis membranes are used to remove accumulated uremic toxins, excess ions and water from the patient via the dialysate, and to supply (deficit) insufficient ions from the dialysate. Dialysis membranes used clinically in the treatment of patients with renal failure account for by far the largest volume of membranes used worldwide; more than 70 million square meters are used a year. Almost all dialyzers now in use are of the hollow-fiber type. A hollow-fiber dialyzer contains a bundle of approximately 10000 hollow fibers, each with an inner diameter of about 200 microm when wet. The membrane thickness is about 20-45 microm, and the length is 160-250 mm. The walls of the hollow fibers function as the dialysis membrane. Various materials, including cellulose-based materials and synthetic polymers, are used for dialysis membranes. This paper reviews blood purification, hemodialysis and dialysis membranes.

  12. Photocatalytic materials and technologies for air purification. (United States)

    Ren, Hangjuan; Koshy, Pramod; Chen, Wen-Fan; Qi, Shaohua; Sorrell, Charles Christopher


    Since there is increasing concern for the impact of air quality on human health, the present work surveys the materials and technologies for air purification using photocatalytic materials. The coverage includes (1) current photocatalytic materials for the decomposition of chemical contaminants and disinfection of pathogens present in air and (2) photocatalytic air purification systems that are used currently and under development. The present work focuses on five main themes. First, the mechanisms of photodegradation and photodisinfection are explained. Second, system designs for photocatalytic air purification are surveyed. Third, the photocatalytic materials used for air purification and their characteristics are considered, including both conventional and more recently developed photocatalysts. Fourth, the methods used to fabricate these materials are discussed. Fifth, the most significant coverage is devoted to materials design strategies aimed at improving the performance of photocatalysts for air purification. The review concludes with a brief consideration of promising future directions for materials research in photocatalysis. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Ionic behavior of treated water at a water purification plant


    Yanagida, Kazumi; Kawahigashi, Tatsuo


    [Abstract] Water at each processing stage in a water purification plant was extracted and analyzed to investigate changes of water quality. Investigations of water at each processing stage at the water purification plant are discussed herein.

  14. Simplified riboprobe purification using translucent straws as gel tubes. (United States)

    Kol, S; Ben-Shlomo, I; Adashi, E Y; Rohan, R M


    Gel purification of radioactive riboprobes enhances the quality of the ribonuclease protection assay. A simple and effective method for riboprobe purification is described. The method uses acrylamide gels in plastic tubes to achieve electrophoretic separation of the RNA polymerase products.

  15. Protein purification protocols [Methods in molecular biology, v. 59

    National Research Council Canada - National Science Library

    Doonan, Shawn


    ... both chemical and molecular methods, and how to dry and store the purified protein. Protein Purification Protocols provides all that is needed to design and carry out a successful purification program...

  16. Purification and characterization of angiotensin-1 converting enzyme

    African Journals Online (AJOL)



    Apr 10, 2013 ... al., 1999), meat (Jang and Lee, 2005) and marine resources (Je et al., ... species that have been blooming on the offshore areas of. Korea, China and ..... Lim et al. 1893. Table 1. Purification yield and ACE-inhibitory activity of each step. Fraction. Purification step. Purification yield (%). IC50 (μg/ml). W/H.

  17. Laboratory technology for hydrogen purification in liquefying installations

    International Nuclear Information System (INIS)

    Avram, I.; David, E.; Dordea, M.; Peculea, M.; Pop, F.; Stanciu, V.; Varzaru, O.; Panu, E.; Curuia, M.; Fron, P.; Balint, I.; Culcer, M.; Roman, T.; Smeureanu, N.


    This paper presents the development of a laboratory installation for purification of the hydrogen to be liquefied. The purification is achieved by the procedure of physical adsorption at low temperature. This procedure implies the use of materials with extensive active surfaces such as activated carbon, molecular sieves and silica gels. The main stages of the purification process are described

  18. Purification of contaminated groundwater by membrane technology

    Energy Technology Data Exchange (ETDEWEB)

    Youn, In Soo; Chung, Chin Ki; Kim, Byoung Gon [Korea Institute of Geology Mining and Materials, Taejon (Korea, Republic of)


    The objective of this study is to apply the membrane separation technology to the purification of contaminated ground water in Korea. Under this scope, the purification was aimed to the drinking water level. The scale of the membrane system was chosen to a small filtration plant for local clean water supplies and/or heavy purifiers for buildings and public uses. The actual conditions of ground water contamination in Korea was surveyed to determine the major components to remove under the drinking water requirements. To set up a hybrid process with membrane methods, conventional purification methods were also investigated for the comparison purpose. The research results are summarized as follows : 1) Contamination of the groundwater in Korea has been found to be widespread across the country. The major contaminant were nitrate, bacteria, and organic chlorides. Some solvents and heavy metals are also supposed to exist in the ground water of industrial complexes, cities, and abandoned mines. 2) The purification methods currently used in public filtration plants appear not to be enough for new contaminants from recent industrial expanding. The advanced purification technologies generally adopted for this problem have been found to be unsuitable due to their very complicated design and operation, and lack of confidence in the purification performance. 3) The reverse osmosis tested with FilmTec FT30 membrane was found to remove nitrate ions in water with over 90 % efficiency. 4) The suitable membrane process for the contaminated groundwater in Korea has been found to be the treatments composed of activated carbon, microfiltration, reverse osmosis or ultrafiltration, and disinfection. The activated carbon treatment could be omitted for the water of low organic contaminants. The microfiltration and the reverse osmosis treatments stand for the conventional methods of filtration plants and the advanced methods for hardly removable components, respectively. It is recommended

  19. Purification and characterization of xylanase from Aspergillus ...

    African Journals Online (AJOL)

    Xylanase was subjected to a three-step purification scheme involving ammonium sulphate precipitation, gel filtration chromatography and anion exchange chromatography. Purity was verified by running the extracted protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and a single band was ...

  20. Expression, purification and characterization of the interferon ...

    Indian Academy of Sciences (India)


    Jan 19, 2012 ... utilizing a single-step affinity purification with an appreciable yield of the highly purified protein. Recombinant RNase L was characterized by SDS-PAGE, immunoblotting and MALDI-TOF analysis. A semi-quantitative agarose-gel-based ribonuclease assay was developed for measuring its 2-5A-dependent ...

  1. Purification and characterization of circulating Onchocerca volvulus ...

    African Journals Online (AJOL)

    ... high antigen titres were separately pooled and subjected to affinity purification using immunosorbent columns prepared using human and rabbit anti-O. volvulus IgG antibodies. Eluates of purified circulating O. volvulus antigens were concentrated, and then the protein contents were determined using the Bradford method.

  2. Purification, characterization of phytase enzyme from Lactobacillus ...

    African Journals Online (AJOL)

    Purification, characterization of phytase enzyme from Lactobacillus plantarum bacteria and determination of its kinetic properties. ... Many of the cereal grains, legumes and oilseeds store phosphorus in phytate form. Phytases can be produced by plants, animals and microorganisms. However, the ones with microbial origin ...

  3. Affinity purification of recombinant human plasminogen activator ...

    African Journals Online (AJOL)

    Purpose: To develop processes for effective isolation and purification of recombinant human plasminogen activator (rhPA) from transgenic rabbit milk. Methods: Immunoaffinity chromatography was selected and improved by a special polyol-responsive monoclonal antibody (PR-mAb). Alteplase was used as immunogen ...

  4. Expression and Purification of Soluble, Biologically Active ...

    African Journals Online (AJOL)

    Expression and Purification of Soluble, Biologically Active Recombinant Dipeptidyl Peptidase 4 (DPP4/CD26/ADAbp) Comprising the Extracellular Domain in the Yeast, Pichia ... Purpose: To investigate Pichia pastoris expression system for producing clinically usable, high-quality dipeptidyl peptidase 4 recombinant protein.

  5. Purification and characterisation of Cyclodextrin glycosyltransferase ...

    African Journals Online (AJOL)



    Jun 5, 2012 ... affinity chromatography were 25.8 and 17.8%, respectively. ... industrial applications according to its characteristic found in the current study. ..... and single step purification of cyclodextrin glycosyltransferase from alkalophilic Bacillus firmus by ion extchange chromatography. Biochem. Eng. J., 39: 510-515.

  6. Partial Purification and Characterization of Extracellular Protease ...

    African Journals Online (AJOL)


    ABSTRACT: Microbial proteases have wide industrial applications and proteases of the lactic acid bacteria (LAB) have received special attention ... Purification of the enzyme by gel filtration chromatography on Sephadex G75 following ammonium .... Gel filtration and ion exchange chromatography. The dialysate was ...

  7. Purification and Characterization of Thermostable Cellulase from ...

    African Journals Online (AJOL)

    CMCase was purified with ion-exchange and gel filtration chromatography. Results: Sequence-based identification of species belonging to the ... industrial application such as biofuel, animal feed industry, paper industry and clarification of fruit juices. Keywords: Thermostable cellulase, Sugarcane bagasse, Purification, ...

  8. Purification And Characterization Of Marine Bacillus Thuringiensis ...

    African Journals Online (AJOL)

    Urease was purified to homogeneity from Bacillus thuringiensis N2 using different purification steps namely, 55% acetone precipitation, DEAE-Sephadex A50 anion exchange column and Sephadex G120-200 gel filtration chromatography. The enzyme was purified 95.27 fold and showed a final specific activity of 10.48 ...

  9. Purification and Characterization of Thermostable Cellulase from ...

    African Journals Online (AJOL)

    ... after incubation at 80 °C for 60 min. Conclusion: Due to its high temperature stability, the purified XM70-CMCase may be useful for industrial application such as biofuel, animal feed industry, paper industry and clarification of fruit juices. Keywords: Thermostable cellulase, Sugarcane bagasse, Purification, Characterization ...

  10. Heparin affinity purification of extracellular vesicles

    NARCIS (Netherlands)

    Balaj, Leonora; Atai, Nadia A.; Chen, Weilin; Mu, Dakai; Tannous, Bakhos A.; Breakefield, Xandra O.; Skog, Johan; Maguire, Casey A.


    Extracellular vesicles (EVs) are lipid membrane vesicles released by cells. They carry active biomolecules including DNA, RNA, and protein which can be transferred to recipient cells. Isolation and purification of EVs from culture cell media and biofluids is still a major challenge. The most widely

  11. Purification and characterization of a thermostable glucoamylase ...

    African Journals Online (AJOL)



    Jun 7, 2010 ... Department of Biology, Faculty of Science and Arts, Adnan Menderes University, 09010, Aydın, Turkey. ... Raw corn starch adsorption of GA was found ...... GP-21 in solid state fermentation. J. Ind. Microbiol. Biotechnol. 22: 622-626. Mamo G, Gessesse A (1999b). Purification and characterization of two.

  12. Isolation, purification and effects of hypoglycemic functional ...

    African Journals Online (AJOL)



    Apr 12, 2012 ... Key words: Hypoglycemic activity, polysaccharides, Inonotus obliquus, isolation, purification. INTRODUCTION. Obesity, high blood glucose and diabetes which are the significant chronic diseases and causes of death in modern society and they are largely caused by diet. Worldwide, scientists have focused ...

  13. Partial purification and characterization of polygalacturonase ...

    African Journals Online (AJOL)

    Partial purification and characterization of polygalacturonase-inhibitor proteins from pearl millet. ... Protein separation of the two peaks by gel electrophoresis showed prominent bands between 29 and 43 kDa, consistent with the molecular weights of the known plant PGIPs. The two PGIP peaks were further studied for their ...

  14. Partial purification and biochemical characterization of acid ...

    African Journals Online (AJOL)

    Mung bean (Vigna radiata) is one of the important crops of the North Eastern Region of India. In the present study, acid phosphatase enzyme was isolated and partially purified from germinated local mung bean seeds. The sequential partial purification process was performed using ammonium sulphate precipitation method.

  15. Purification of functionalized DNA origami nanostructures. (United States)

    Shaw, Alan; Benson, Erik; Högberg, Björn


    The high programmability of DNA origami has provided tools for precise manipulation of matter at the nanoscale. This manipulation of matter opens up the possibility to arrange functional elements for a diverse range of applications that utilize the nanometer precision provided by these structures. However, the realization of functionalized DNA origami still suffers from imperfect production methods, in particular in the purification step, where excess material is separated from the desired functionalized DNA origami. In this article we demonstrate and optimize two purification methods that have not previously been applied to DNA origami. In addition, we provide a systematic study comparing the purification efficacy of these and five other commonly used purification methods. Three types of functionalized DNA origami were used as model systems in this study. DNA origami was patterned with either small molecules, antibodies, or larger proteins. With the results of our work we aim to provide a guideline in quality fabrication of various types of functionalized DNA origami and to provide a route for scalable production of these promising tools.

  16. Purification, Characterization and Antibacterial Mechanism of ...

    African Journals Online (AJOL)

    Purpose: To carry out the extraction, purification and biological characterization, and assess the antibacterial activity of bacteriocin from Lactobacillus acidophilus XH1. Methods: Chloroform extraction method was used for bacteriocin extraction while characterization of bacteriocin was carried out by flat-dug well agar ...

  17. Rapid cloning and purification of proteins: gateway vectors for protein purification by self-cleaving tags. (United States)

    Gillies, Alison R; Hsii, Judy F; Oak, Seachol; Wood, David W


    We have combined Invitrogen's Gateway cloning technology with self-cleaving purification tags to generate a new system for rapid production of recombinant protein products. To accomplish this, we engineered our previously reported DeltaI-CM cleaving intein to include a Gateway cloning recognition sequence, and demonstrated that the resulting Gateway-competent intein is unaffected. This intein can therefore be used in several previously reported purification methods, while at the same time being compatible with Gateway cloning. We have incorporated this intein into a set of Gateway vectors, which include self-cleaving elastin-like polypeptide (ELP), chitin binding domain (CBD), phasin (polyhydroxybutyrate-binding), or maltose binding domain (MBD) tags. These vectors were verified by Gateway cloning of TEM-1 beta-lactamase and Escherichia coli catalase genes, and the expressed target proteins were purified using the four methods encoded on the vectors. The purification methods were unaffected by replacing the DeltaI-CM intein with the Gateway intein. It was observed that some purification methods were more appropriate for each target than others, suggesting utility of this technology for rapid process identification and optimization. The modular design of the Gateway system and intein purification method suggests that any tag and promoter can be trivially added to this system for the development of additional expression vectors. This technology could greatly facilitate process optimization, allowing several targets and methods to be tested in a high-throughput manner.


    DEFF Research Database (Denmark)

    Nielsen, Joan Maj; Dahi, Elian


    Purification of fluoride contaminated magadi is studied using bone char sorption and calcium precipitation. The bone char treatment is found to be workable both in columns and in batches where the magadi is dissolved in water prior to treatment. The concentrations in the solutions were 89 g magadi....../L and 95 and 400 mg F/L respectively in natural and synthetic solutions. The fluoride removal capacities observed were 4.6 mg F/g bone char for the column system and 2.7 mg F/g bone char for the batch system in case of synthetic magadi solution. It is however concluded that the batch system is the best...... treatment method. A procedure for purification of fluoride contaminated magadi at household level is described....

  19. The Viability of Photocatalysis for Air Purification

    Directory of Open Access Journals (Sweden)

    Stephen O. Hay


    Full Text Available Photocatalytic oxidation (PCO air purification technology is reviewed based on the decades of research conducted by the United Technologies Research Center (UTRC and their external colleagues. UTRC conducted basic research on the reaction rates of various volatile organic compounds (VOCs. The knowledge gained allowed validation of 1D and 3D prototype reactor models that guided further purifier development. Colleagues worldwide validated purifier prototypes in simulated realistic indoor environments. Prototype products were deployed in office environments both in the United States and France. As a result of these validation studies, it was discovered that both catalyst lifetime and byproduct formation are barriers to implementing this technology. Research is ongoing at the University of Connecticut that is applicable to extending catalyst lifetime, increasing catalyst efficiency and extending activation wavelength from the ultraviolet to the visible wavelengths. It is critical that catalyst lifetime is extended to realize cost effective implementation of PCO air purification.

  20. Nanotechnology for water treatment and purification

    CERN Document Server

    Apblett, Allen


    This book describes the latest progress in the application of nanotechnology for water treatment and purification. Leaders in the field present both the fundamental science and a comprehensive overview of the diverse range of tools and technologies that have been developed in this critical area. Expert chapters present the unique physicochemical and surface properties of nanoparticles and the advantages that these provide for engineering applications that ensure a supply of safe drinking water for our growing population. Application areas include generating fresh water from seawater, preventing contamination of the environment, and creating effective and efficient methods for remediation of polluted waters. The chapter authors are leading world-wide experts in the field with either academic or industrial experience, ensuring that this comprehensive volume presents the state-of-the-art in the integration of nanotechnology with water treatment and purification. Covers both wastewater and drinking water treatmen...

  1. Nanomaterials and Water Purification: Opportunities and Challenges

    International Nuclear Information System (INIS)

    Savage, Nora; Diallo, Mamadou S.


    Advances in nanoscale science and engineering suggest that many of the current problems involving water quality could be resolved or greatly ameliorated using nanosorbents, nanocatalysts, bioactive nanoparticles, nanostructured catalytic membranes and nanoparticle enhanced filtration among other products and processes resulting from the development of nanotechnology. Innovations in the development of novel technologies to desalinate water are among the most exciting and promising. Additionally, nanotechnology-derived products that reduce the concentrations of toxic compounds to sub-ppb levels can assist in the attainment of water quality standards and health advisories. This article gives an overview of the use of nanomaterials in water purification. We highlight recent advances on the development of novel nanoscale materials and processes for treatment of surface water, groundwater and industrial wastewater contaminated by toxic metal ions, radionuclides, organic and inorganic solutes, bacteria and viruses. In addition, we discuss some challenges associated with the development of cost effective and environmentally acceptable functional nanomaterials for water purification

  2. Nanomaterials and Water Purification: Opportunities and Challenges (United States)

    Savage, Nora; Diallo, Mamadou S.


    Advances in nanoscale science and engineering suggest that many of the current problems involving water quality could be resolved or greatly ameliorated using nanosorbents, nanocatalysts, bioactive nanoparticles, nanostructured catalytic membranes and nanoparticle enhanced filtration among other products and processes resulting from the development of nanotechnology. Innovations in the development of novel technologies to desalinate water are among the most exciting and promising. Additionally, nanotechnology-derived products that reduce the concentrations of toxic compounds to sub-ppb levels can assist in the attainment of water quality standards and health advisories. This article gives an overview of the use of nanomaterials in water purification. We highlight recent advances on the development of novel nanoscale materials and processes for treatment of surface water, groundwater and industrial wastewater contaminated by toxic metal ions, radionuclides, organic and inorganic solutes, bacteria and viruses. In addition, we discuss some challenges associated with the development of cost effective and environmentally acceptable functional nanomaterials for water purification.

  3. Development of partitioning process: purification of DIDPA

    International Nuclear Information System (INIS)

    Watanabe, Masayuki; Morita, Yasuji; Kubota, Masumitsu


    The partitioning process has developed and demonstrated that the solvent extraction with diisodecylphosphoric acid (DIDPA) can successfully separate transuranium elements from a high-level liquid waste. In the solvent extraction, DIDPA is decomposed by radiolysis and hydrolysis. The main degradation product is monoisodecyl phosphoric acid (MIDPA). Ethylene glycol has been used for removing the product by a solvent extraction method. However this method has two drawbacks that two phases separate slowly and the used ethylene glycol is not regeneratable. First it was found that the addition of acetone or methanol with 20 volume % improved the phase separation. Then a new purification method was developed by using an aqueous solution of methanol or acetone. The new purification method is as excellent as the ethylene glycol method for the removal of MIDPA. (author)

  4. Using of Mineral Recourses for Water Purification

    International Nuclear Information System (INIS)

    Tumanova, I.V.; Nazarenko, O.B.; Anna, Yu.


    Pollution of surface waters results in necessity of underground waters using for drinking. Underground waters are characterized by the high quantity of heavy metals salts. This led to development of methods reducing the concentration of the metal salts in water. Wide spread occurrence, cheapness and high sorption properties of nature minerals allow to consider them as perspective sorbents for different impurities extraction, including dissoluble compounds of heavy metals. Reachable purification efficiency with mineral resources use for the moment satisfies sanitary indexes and standards presenting to portable water in Russia. In given material there are presented the results of research of artificial sorbent and certain minerals sorption characteristics, which are typical for West Siberia. For purification quality improvement from Fe and Mn ions there are suggested to use the method of boiling bed.

  5. Entanglement of purification: from spin chains to holography (United States)

    Nguyen, Phuc; Devakul, Trithep; Halbasch, Matthew G.; Zaletel, Michael P.; Swingle, Brian


    Purification is a powerful technique in quantum physics whereby a mixed quantum state is extended to a pure state on a larger system. This process is not unique, and in systems composed of many degrees of freedom, one natural purification is the one with minimal entanglement. Here we study the entropy of the minimally entangled purification, called the entanglement of purification, in three model systems: an Ising spin chain, conformal field theories holographically dual to Einstein gravity, and random stabilizer tensor networks. We conjecture values for the entanglement of purification in all these models, and we support our conjectures with a variety of numerical and analytical results. We find that such minimally entangled purifications have a number of applications, from enhancing entanglement-based tensor network methods for describing mixed states to elucidating novel aspects of the emergence of geometry from entanglement in the AdS/CFT correspondence.

  6. Purification of crude biodiesel using dry washing and membrane technologies

    Directory of Open Access Journals (Sweden)

    I.M. Atadashi


    Full Text Available Purification of crude biodiesel is mandatory for the fuel to meet the strict international standard specifications for biodiesel. Therefore, this paper carefully analyzed recently published literatures which deal with the purification of biodiesel. As such, dry washing technologies and the most recent membrane biodiesel purification process have been thoroughly examined. Although purification of biodiesel using dry washing process involving magnesol and ion exchange resins provides high-quality biodiesel fuel, considerable amount of spent absorbents is recorded, besides the skeletal knowledge on its operating process. Further, recent findings have shown that biodiesel purification using membrane technique could offer high-quality biodiesel fuel with less wastewater discharges. Thus, both researchers and industries are expected to benefit from the development of membrane technique in purifying crude biodiesel. As well biodiesel purification via membranes has been shown to be environmentally friendly. For these reasons, it is important to explore and exploit membrane technology to purify crude biodiesel.

  7. Conductive diamond electrodes for water purification

    Directory of Open Access Journals (Sweden)

    Carlos Alberto Martínez-Huitle


    Full Text Available Nowadays, synthetic diamond has been studied for its application in wastewater treatment, electroanalysis, organic synthesis and sensor areas; however, its use in the water disinfection/purification is its most relevant application. The new electrochemistry applications of diamond electrodes open new perspectives for an easy, effective, and chemical free water treatment. This article highlights and summarizes the results of a selection of papers dealing with electrochemical disinfection using synthetic diamond films.

  8. Propagation and Purification of Baculovirus oryctes Huger

    Directory of Open Access Journals (Sweden)

    Susamto Somowiyarjo


    Full Text Available An isolate of Baculovirus oryctes, a possible biological control agent for coconut beetle (Oryctes rhinoceros Huger from East Java was propagated and purified. The virus could be transmitted by feeding the imago with 10% sucrose containing virus from homogenate of infected beetles. Effectivity of virus to 9 healthy females by sexual copulation. Virus be succesfully purified by a method of Payne. Key words: Baculovirus oryctes, transmission, purification


    African Journals Online (AJOL)


    cristallinité, son complexe avec la lignine et les autres composés qui lui sont associés. (Arunik et al., 1988). Pour pouvoir hydrolyser les polymères naturels, nous avons entrepris l'extraction, la purification, l'homogénéisation et la déter- mination des propriétés physico-chimiques de deux cellulases des ouvriers du termite ...

  10. Overview of the purification of recombinant proteins. (United States)

    Wingfield, Paul T


    When the first version of this unit was written in 1995, protein purification of recombinant proteins was based on a variety of standard chromatographic methods and approaches, many of which were described and mentioned throughout Current Protocols in Protein Science. In the interim, there has been a shift toward an almost universal usage of the affinity or fusion tag. This may not be the case for biotechnology manufacture where affinity tags can complicate producing proteins under regulatory conditions. Regardless of the protein expression system, questions are asked as to which and how many affinity tags to use, where to attach them in the protein, and whether to engineer a self-cleavage system or simply leave them on. We will briefly address some of these issues. Also, although this overview focuses on E.coli, protein expression and purification, other commonly used expression systems are mentioned and, apart from cell-breakage methods, protein purification methods and strategies are essentially the same. Copyright © 2015 John Wiley & Sons, Inc.

  11. Rotating Reverse-Osmosis for Water Purification (United States)

    Lueptow, RIchard M.


    A new design for a water-filtering device combines rotating filtration with reverse osmosis to create a rotating reverse- osmosis system. Rotating filtration has been used for separating plasma from whole blood, while reverse osmosis has been used in purification of water and in some chemical processes. Reverse- osmosis membranes are vulnerable to concentration polarization a type of fouling in which the chemicals meant not to pass through the reverse-osmosis membranes accumulate very near the surfaces of the membranes. The combination of rotating filtration and reverse osmosis is intended to prevent concentration polarization and thereby increase the desired flux of filtered water while decreasing the likelihood of passage of undesired chemical species through the filter. Devices based on this concept could be useful in a variety of commercial applications, including purification and desalination of drinking water, purification of pharmaceutical process water, treatment of household and industrial wastewater, and treatment of industrial process water. A rotating filter consists of a cylindrical porous microfilter rotating within a stationary concentric cylindrical outer shell (see figure). The aqueous suspension enters one end of the annulus between the inner and outer cylinders. Filtrate passes through the rotating cylindrical microfilter and is removed via a hollow shaft. The concentrated suspension is removed at the end of the annulus opposite the end where the suspension entered.

  12. Efficacy of organic matter and some bio-inoculants for the management of root-knot nematode infesting tomato

    Directory of Open Access Journals (Sweden)

    Neha Khan


    Full Text Available Efficiency of an organic matter like Tagetes erecta and bioinoculants Azotobacter chroococcum and Glomus fasciculatum was investigated in tomato cultivar ‘Pusa Ruby’ when inoculated individually as well as concomitantly for the management of the root-knot nematode, Meloidogyne incognita in terms of growth parameters such as plant length, fresh and dry weights, chlorophyll content, per cent pollen fertility and mycorrhization. Greatest reduction in the numbers of second-stage juveniles in soil, number of root-galls, egg-masses and nematode multiplication was recorded with combined application of T. erecta and bio-inoculants A. chroococcum and G. fasciculatum as compared to untreated control and other treatments. Similarly, the greatest improvement in the plant growth and biomass of tomato was noted in the same treatments. However, individual inoculation of these bio-inoculants and organic fertilizers also showed significant enhancement but was less as compared to combined treatment. A. chroococcum was found most effective against disease incidence followed by G. fasciculatum and T. erecta. Parameters like NP and K contents were significantly enhanced in those plants which received combined treatments of organic matter and bio-inoculants. Azotobacter was found more efficacious against nematodes than Glomus fasciculatum. Organic matter also influenced the activity of bio-inoculants, more with the Azotobacter than G. fasciculatum. DOI: International Journal of Environment Vol.4(2 2015: 206-220

  13. Mobile sequences in the pyruvate dehydrogenase complex, the E2 component, the catalytic domain and the 2-oxogluturate dehydrogenase complex of Azotobacter vinelandii, as detected by 600 MHz 1H-NMR spectroscopy

    International Nuclear Information System (INIS)

    Hanemaaijer, R.; Vervoort, J.; Westphal, A.H.; Kok, A. de.; Veeger, C.


    600 MHz 1 H-NMR spectroscopy demonstrates that the pyruvate dehydrogenase complex of Azotobacter vinelandii contains regions of the polypeptide chain with intramolecular mobility. This mobility is located in the E 2 component and can probably be ascribed to alanine-proline-rich regions that link the lipoyl sibdiomains to each other as well as to the E 1 and E 3 binding domain. In the catalytic domain of E 2 which is thought to form a compact, rigid core, also conformational flexibility is observed. It is conceivable that the N-terminal region of the catalytic domain, which contains many alanine residues, is responsible for the observed mobility. In the low-field region of the 1 H-NMR spectrum of E 2 specific resonances are found, which can be ascribed to mobile phenylalanine, histidine and/or tyrosine residues which are located in the E 1 and E 3 binding domain that links the lipoyl domain to the catalytic domain. In the 1 H-NMR spectrum of the intact complex, these resonances cannot be observed, indicating a decreased mobility of the E 1 and E 3 binding domain. (author). 24 refs.; 2 figs

  14. A Scintillator Purification System for the Borexino Solar Neutrino Detector


    Benziger, J.; Cadonati, L.; Calaprice, F.; Chen, M.; Corsi, A.; Dalnoki-Veress, F.; Fernholz, R.; Ford, R.; Galbiati, C.; Goretti, A.; Harding, E.; Ianni, Aldo; Ianni, Andrea; Kidner, S.; Leung, M.


    Purification of the 278 tons of liquid scintillator and 889 tons of buffer shielding for the Borexino solar neutrino detector was performed with a system that combined distillation, water extraction, gas stripping and filtration. The purification of the scintillator achieved unprecedented low backgrounds for the large scale liquid scintillation detector. This paper describes the principles of operation, design, construction and commissioning of the purification system, and reviews the require...

  15. Nanostructured Catalytic Reactors for Air Purification, Phase I (United States)

    National Aeronautics and Space Administration — This SBIR Phase I project proposes the development of lightweight compact nanostructured catalytic reactors for air purification from toxic gaseous organic...

  16. Compressorless Gas Storage and Regenerative Hydrogen Purification, Phase I (United States)

    National Aeronautics and Space Administration — Microwave regenerative sorption media gas storage/delivery techniques are proposed to address both compressed gas management and hydrogen purification requirements...

  17. Nanostructured Catalytic Reactors for Air Purification, Phase II (United States)

    National Aeronautics and Space Administration — This SBIR Phase II project proposes the development of lightweight compact nanostructured catalytic reactors for air purification from toxic gaseous organic...

  18. Dense Medium Plasma Water Purification Reactor (DMP WaPR), Phase I (United States)

    National Aeronautics and Space Administration — The Dense Medium Plasma Water Purification Reactor offers significant improvements over existing water purification technologies used in Advanced Life Support...

  19. Effects of organic and biological fertilizers on fruit yield and essential oil of sweet fennel (Foeniculum vulgare var. dulce)

    Energy Technology Data Exchange (ETDEWEB)

    Moradi, R.; Rezvani Moghaddam, P.; Nasiri Mahallati, M.; Nezhadali, A.


    In order to evaluate the effects of different organic and biological fertilizers on quantity and quality of fennel essential oil, an experiment was conducted in a completely randomized block design with three replications. The experimental treatments included two organic (compost and vermicompost) and two biological (Pseudomonas putida and Azotobacter chroococcum) fertilizers, their all twin combinations (Ps. putida + A. chroococcum, Ps. putida + compost, Ps. putida + vermicompost, A. chroococcum + compost, A. chroococcum + vermicompost and compost + vermicompost) and control (non fertilized). There were significant differences between treatments in terms of seed essential oil percentage, essential oil yield; anethole, fenchone, limonene and straggle content in seed essential oil. Results showed that the highest and the lowest percentages of essential oil were obtained in control (2.9%) and A. chroococcum + vermicompost (2.2%) treatments, respectively. The highest essential oil yield (29.9 L ha{sup -}1) and anethole content of essential oil (69.7%) and the lowest contents of fenchone (6.14%), limonene (4.84%) and estragole (2.78%) in essential oil were obtained in compost + vermicompost treatment. It seems that compost + vermicompost treatment compared to other treatments supplied the highest equilibrium of nutrients and water in the root zone of sweet fennel which is led to increasing the anethole content, there upon, decreasing other compounds. Essential oil yield and percentage of anethole content in essential oil were significantly higher in all organic and biological treatments compared with control. (Author) 43 refs.

  20. A scintillator purification system for the Borexino solar neutrino detector

    Energy Technology Data Exchange (ETDEWEB)

    Benziger, J. [Chemical Engineering Department, Princeton University, Princeton, NJ 08544 (United States)], E-mail:; Cadonati, L.; Calaprice, F.; Chen, M.; Corsi, A.; Dalnoki-Veress, F.; Fernholz, R.; Ford, R.; Galbiati, C. [Physics Department, Princeton University, Princeton, NJ 08544 (United States); Goretti, A. [INFN, Laboratori Nazionale di Gran Sasso (Italy); Harding, E. [Physics Department, Princeton University, Princeton, NJ 08544 (United States); Ianni, Aldo [INFN, Laboratori Nazionale di Gran Sasso (Italy); Ianni, Andrea; Kidner, S.; Leung, M.; Loeser, F.; McCarty, K.; McKinsey, D.; Nelson, A.; Pocar, A. [Physics Department, Princeton University, Princeton, NJ 08544 (United States)] (and others)


    Purification of the 278 tons of liquid scintillator and 889 tons of buffer shielding for the Borexino solar neutrino detector is performed with a system that combines distillation, water extraction, gas stripping, and filtration. This paper describes the principles of operation, design, and construction of that purification system, and reviews the requirements and methods to achieve system cleanliness and leak-tightness.

  1. Purification and Characterization of Lipase from Aspergillus flavus ...

    African Journals Online (AJOL)


    Abstract. Lipase from Aspergillus flavus was purified in a single step purification using MnFeO4 magnetic nano particles to achieve a 20.53- fold purification with specific activity of. 11.29 U/mg and a 59% recovery yield. SDS-PAGE of lipase showed a single pure band with corresponding molecular weight of 35 kDa.

  2. A scintillator purification system for the Borexino solar neutrino detector (United States)

    Benziger, J.; Cadonati, L.; Calaprice, F.; Chen, M.; Corsi, A.; Dalnoki-Veress, F.; Fernholz, R.; Ford, R.; Galbiati, C.; Goretti, A.; Harding, E.; Ianni, Aldo; Ianni, Andrea; Kidner, S.; Leung, M.; Loeser, F.; McCarty, K.; McKinsey, D.; Nelson, A.; Pocar, A.; Salvo, C.; Schimizzi, D.; Shutt, T.; Sonnenschein, A.


    Purification of the 278 tons of liquid scintillator and 889 tons of buffer shielding for the Borexino solar neutrino detector is performed with a system that combines distillation, water extraction, gas stripping, and filtration. This paper describes the principles of operation, design, and construction of that purification system, and reviews the requirements and methods to achieve system cleanliness and leak-tightness.

  3. New Combined Electron-Beam Methods of Wastewater Purification

    International Nuclear Information System (INIS)

    Pikaev, A.K.; Makarov, I.E.; Ponomarev, A.V.; Kartasheva, L.I.; Podzorova, E.A.; Chulkov, V.N.; Han, B.; Kim, D.K.


    The paper is a brief review of the results obtained with the participation of the authors from the study on combined electron-beam methods for purification of some wastewaters. The data on purification of wastewaters containing dyes or hydrogen peroxide and municipal wastewater in the aerosol flow are considered

  4. Surface processes during purification of InP quantum dots

    Directory of Open Access Journals (Sweden)

    Natalia Mordvinova


    Full Text Available Recently, a new simple and fast method for the synthesis of InP quantum dots by using phosphine as phosphorous precursor and myristic acid as surface stabilizer was reported. Purification after synthesis is necessary to obtain samples with good optical properties. Two methods of purification were compared and the surface processes which occur during purification were studied. Traditional precipitation with acetone is accompanied by a small increase in photoluminescence. It occurs that during the purification the hydrolysis of the indium precursor takes place, which leads to a better surface passivation. The electrophoretic purification technique does not increase luminescence efficiency but yields very pure quantum dots in only a few minutes. Additionally, the formation of In(OH3 during the low temperature synthesis was explained. Purification of quantum dots is a very significant part of postsynthetical treatment that determines the properties of the material. But this subject is not sufficiently discussed in the literature. The paper is devoted to the processes that occur at the surface of quantum dots during purification. A new method of purification, electrophoresis, is investigated and described in particular.

  5. Comparative study of peroxidase purification from apple and orange ...

    African Journals Online (AJOL)

    This paper reports the isolation and purification of peroxidase from low cost material; moreover, no significant work has been done on the isolation and purification of peroxidase from such cost effective sources (apple and orange seeds). Peroxidases had attracted considerable interest in recent years because of their ...

  6. Overview of the recombinant proteins purification by affinity tags and ...

    African Journals Online (AJOL)

    From protein within isolation process which the same matter increases labor costs further and prevents application of these tags in industrial scale. Therefore proper replacement is emphasized for enzymatic removal of purification tags. Keywords: protein purification; recombinant proteins; self-cleavable tags; Intein tags; ...

  7. Effect of Purification Procedures on DIF Analysis in IRTPRO (United States)

    Fikis, David R. J.; Oshima, T. C.


    Purification of the test has been a well-accepted procedure in enhancing the performance of tests for differential item functioning (DIF). As defined by Lord, purification requires reestimation of ability parameters after removing DIF items before conducting the final DIF analysis. IRTPRO 3 is a recently updated program for analyses in item…

  8. Purification of cadmium up to 5N+ by vacuum distillation

    Indian Academy of Sciences (India)


    Other areas of applications include telecommunications, electric vehicle and remote area storage systems. The problem connected with profound purification and characterization of the materials that are used for the synthesis of electronic materials remains critical. The advantage of methods providing efficient purification.

  9. Optimization of laboratory scale production and purification of ...

    African Journals Online (AJOL)

    Microcystin content is however highly variable and optimised culture conditions are essential to produce viable yields of microcystin for purification. We describe the optimization of culture conditions and evaluation of various purification methods to enhance the yield of microcystin from laboratory scale culture.

  10. New matrices for the purification of pectinases by affinity chromatography. (United States)

    Lobarzewski, J; Fiedurek, J; Ginalska, G; Wolski, T


    Polygalacturonic acid was used as a ligand in the affinity technique for pectinases purification from the filtrate of Aspergillus niger 71 culture. For this purpose four matrices were examined, namely, alkylamine controlled porous glass (CPG), alkylamine silica gel as well as keratin or polyamide coated silica gel. Good results of pectinase purification was obtained on silanized CPG or keratin coated silica gel supports.

  11. Necessity of purification during bacterial DNA extraction with environmental soils

    Directory of Open Access Journals (Sweden)

    Hyun Jeong Lim


    Full Text Available Complexity and heterogeneity of soil samples have often implied the inclusion of purification steps in conventional DNA extraction for polymerase chain reaction (PCR assays. Unfortunately the purification steps are also time and labor intensive. Therefore the necessity of DNA purification was re-visited and investigated for a variety of environmental soil samples that contained various amounts of PCR inhibitors. Bead beating and centrifugation was used as the baseline (without purification method for DNA extraction. Its performance was compared with that of conventional DNA extraction kit (with purification. The necessity criteria for DNA purification were established with environmental soil samples. Using lysis conditions at 3000 rpm for 3 minutes with 0.1 mm glass beads, centrifugation time of 10 minutes and 1:10 dilution ratio, the baseline method outperformed conventional DNA extraction on cell seeded sand samples. Further investigation with PCR inhibitors (i.e., humic acids, clay, and magnesium [Mg] showed that sand samples containing less than 10 μg/g humic acids and 70% clay may not require purifications. Interestingly, the inhibition pattern of Mg ion was different from other inhibitors due to the complexation interaction of Mg ion with DNA fragments. It was concluded that DNA extraction method without purification is suitable for soil samples that have less than 10 μg/g of humic acids, less than 70% clay content and less than 0.01% Mg ion content.

  12. Purification and characterization of a protease from Thermophilic ...

    African Journals Online (AJOL)

    The purification and characterization of a thermophilic neutral protease from Thermophilic bacillus strain HS08, originally isolated from a soil sample collected from the Tulufan Crater of China, is presented in this paper. The purification steps included ammonium sulfate precipitation, with columns of DEAE-Sepharose anion ...

  13. Purification and characterization of a novel extracellular chitinase ...

    African Journals Online (AJOL)

    The purification and characterization of a new thermophilic chitinase from thermophilic Bacillus sp. HU1, originally isolated from a soil sample collected from hot spring of XinJiang Province, China, is presented in this paper. The purification steps included ammonium sulfate precipitation, with columns of DEAE-Sepharose ...

  14. Purification and fluorescent labeling of the human serotonin transporter

    DEFF Research Database (Denmark)

    Rasmussen, Søren G F; Gether, Ulrik


    To establish a purification procedure for the human serotonin transporter (hSERT) we expressed in Sf9 insect cells an epitope-tagged version of the transporter containing a FLAG epitope at the N-terminus and a polyhistidine tail at the C-terminus (FLAG-hSERT-12H). For purification, the transporter...

  15. One-step purification of E. coli elongation factor Tu

    DEFF Research Database (Denmark)

    Knudsen, Charlotte Rohde; Clark, Brian F. C.; Degn, B


    The tuf A gene, encoding the E. coli elongation factor Tu, was cloned in the pGEX gene fusion system. Upon expression EF-Tu is fused to glutathione-S-transferase serving as a purification handle with affinity for glutathione immobilised on agarose. This allows purification of EF-Tu in a one...

  16. Ribonucleoprotein purification and characterization using RNA Mango. (United States)

    Panchapakesan, Shanker Shyam S; Ferguson, Matthew L; Hayden, Eric J; Chen, Xin; Hoskins, Aaron A; Unrau, Peter J


    The characterization of RNA-protein complexes (RNPs) is a difficult but increasingly important problem in modern biology. By combining the compact RNA Mango aptamer with a fluorogenic thiazole orange desthiobiotin (TO1-Dtb or TO3-Dtb) ligand, we have created an RNA tagging system that simplifies the purification and subsequent characterization of endogenous RNPs. Mango-tagged RNP complexes can be immobilized on a streptavidin solid support and recovered in their native state by the addition of free biotin. Furthermore, Mango-based RNP purification can be adapted to different scales of RNP isolation ranging from pull-down assays to the isolation of large amounts of biochemically defined cellular RNPs. We have incorporated the Mango aptamer into the S. cerevisiae U1 small nuclear RNA (snRNA), shown that the Mango-snRNA is functional in cells, and used the aptamer to pull down a U1 snRNA-associated protein. To demonstrate large-scale isolation of RNPs, we purified and characterized bacterial RNA polymerase holoenzyme (HE) in complex with a Mango-containing 6S RNA. We were able to use the combination of a red-shifted TO3-Dtb ligand and eGFP-tagged HE to follow the binding and release of the 6S RNA by two-color native gel analysis as well as by single-molecule fluorescence cross-correlation spectroscopy. Together these experiments demonstrate how the Mango aptamer in conjunction with simple derivatives of its flurophore ligands enables the purification and characterization of endogenous cellular RNPs in vitro. © 2017 Panchapakesan et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  17. Biopharmaceuticals from microorganisms: from production to purification

    Directory of Open Access Journals (Sweden)

    Angela Faustino Jozala

    Full Text Available ABSTRACT The use of biopharmaceuticals dates from the 19th century and within 5-10 years, up to 50% of all drugs in development will be biopharmaceuticals. In the 1980s, the biopharmaceutical industry experienced a significant growth in the production and approval of recombinant proteins such as interferons (IFN α, β, and γ and growth hormones. The production of biopharmaceuticals, known as bioprocess, involves a wide range of techniques. In this review, we discuss the technology involved in the bioprocess and describe the available strategies and main advances in microbial fermentation and purification process to obtain biopharmaceuticals.

  18. Phase equilibria at the Zirconium metal purification

    International Nuclear Information System (INIS)

    Dwiretnani-Sudjoko; Busron-Masduki; Sunardjo; Budi-Sulistyo


    It was investigated the research in the purification of zirconium metal, which was results from the reduction process, by adding heat in the vacuum environment. The process was done in batch in the stainless steel reactor, equiped with vacuum pump and electric heater. The investigated variable were process temperature and pressure. From this research it was obtained that equilibrium constant for MgCl 2 and Mg were expressed in the equation K M g C l 2 = 0.9011 P 1 .3779 1.06552 T and K M g = 6.0115P + 1.35256T - 6.93912

  19. Purification of Argon from a Diluted Stream

    Energy Technology Data Exchange (ETDEWEB)

    Montanari, D. [Princeton U.; Alton, A.; Back, H. O. [Fermilab; Davis, R. P. [Fermilab; Galbiati, C. [Princeton U.; Hardin, K. L. [Fermilab; Jaskierny, W. F. [Fermilab; Kendziora, C. L. [Fermilab; Loer, B. [Princeton U.; Miner, W. E. [Fermilab; Mosteiro, P. J. [Princeton U.; Pordes, S. H. [Fermilab; Subbarao, P. [Princeton U.


    We report on the design, performance and commissioning of a cryogenic distillation column for low radioactivity underground argon purification that has been constructed at Fermi National Accelerator Laboratory. The plant accepts a mixture of argon, helium, and nitrogen with low argon concentration and is designed to return pure argon with a nitrogen contamination less than 10 ppm. During the commissioning, the distillation column in a continuous mode produced argon 99.9% pure. After running in batch mode, the argon purity was increased to 99.95%, with 500 ppm of nitrogen remaining. The argon production rate was about 1 kg/day.

  20. RNA purification by preparative polyacrylamide gel electrophoresis. (United States)

    Petrov, Alexey; Wu, Tinghe; Puglisi, Elisabetta Viani; Puglisi, Joseph D


    Preparative polyacrylamide gel electrophoresis (PAGE) is a powerful tool for purifying RNA samples. Denaturing PAGE allows separation of nucleic acids that differ by a single nucleotide in length. It is commonly used to separate and purify RNA species after in vitro transcription, to purify naturally occurring RNA variants such as tRNAs, to remove degradation products, and to purify labeled RNA species. To preserve RNA integrity following purification, RNA is usually visualized by UV shadowing or stained with ethidium bromide or SYBR green dyes. © 2013 Elsevier Inc. All rights reserved.

  1. Radionuclide characterization of spent purification filters

    International Nuclear Information System (INIS)

    Sinclair, A.N.; Spooner, P.


    A remote handling facility is described which will aid in the assessment of the radionuclide mix and total activity content of spent purification filters. The facility will also dry filters as part of their preparation for off-site shipment. One possible system design, consisting of a robot with five degrees of freedom, a shielded Ge-Li detector, and a vacuum drying system, is described. Benefits could include reduced operating and waste storage or burial costs, feedback on filter performance, and an improved understanding of environmental protection requirements

  2. CAREM-25. Purification and volume control system

    International Nuclear Information System (INIS)

    Acosta, Eduardo; Carlevaris, Rodolfo; Patrignani, Alberto; Chocron, Mauricio; Goya, Hector E.; Ortega, Daniel A.; Ramilo, Lucia B.


    The purification and volume control system has the following main functions: water level control inside reactor pressure vessel (RPR) in all the reactor operational modes, pressure control when the reactor operates in solid state, and maintenance of radiological, physical and chemical parameters of primary water. In case of Hot Shutdown operational mode and also after Scram the system is capable of extraction of nuclear decay heat. The design of the system is in accordance with the Requirements of ANSI/ ANS 51.1; 58.11 and 56.2 standards. (author)

  3. Stimulation of the natural self-purification of soil

    Energy Technology Data Exchange (ETDEWEB)

    Moeller, F.


    Connected with the determination of the sanitary norm of cadmium in soil, examinations were performed to appoint the self-purification index. This index is the highest permissible concentration of a noxious chemical substance in soil not yet injuring the irreversible process of natural self-purification. As criterions of the self-purification were selected normal and potential respiration, quantity of bacteria, actinomyceta, fungi, proteolytic, nitrifying, and cellulolytic micro-organisms, inhibition of Bacillus subtilis and Phormidium autumnale, survival and increasing ability of Escherichia coli, Streptococcus faecalis and Salmonella give. Among the influence of various concentrations of cadmium, the self-purification index of 500 mg Cd++/kg was determined. Offered are methodical improvements for the determination of self-purification indices.

  4. An environment friendly engineeredAzotobactercan replace substantial amount of urea fertilizer and yet sustain same wheat yield. (United States)

    Bageshwar, Umesh K; Srivastava, Madhulika; Pardha-Saradhi, Pedisetty; Paul, Sangeeta; Gothandapani, Sellamuthu; Jaat, Ranjeet S; Shankar, Prabha; Yadav, Rajbir; Biswas, Dipak R; Kumar, Polumetla A; Padaria, Jasdeep C; Mandal, Pranab K; Annapurna, Kannepalli; Das, Hirendra K


    In our endeavor to improve upon nitrogen fixation efficiency of a soil diazotroph that would be unaffected by synthetic nitrogenous fertilizers, we have deleted a part of the negative regulatory gene nifL and constitutively expressed the positive regulatory gene nifA in the chromosome of Azotobacter chroococcum CBD15, a strain isolated from the local field soil. No antibiotic resistance gene or other foreign gene was there in the chromosome of the engineered strain. Wheat seeds inoculated with this engineered strain, which we have named Azotobacter chroococcum HKD15, were tested for three years in pots and one year in the field. Yield of wheat was enhanced by ∼60% due to inoculation of seeds by A. chroococcum HKD15 in the absence of any urea application. Ammonium only marginally affected acetylene reduction by the engineered Azotobacter strain. When urea was also applied, the same wheat yield could be sustained by using seeds inoculated with A. chroococcum HKD15 and using ∼85kg less urea (∼40kg less nitrogen) than the usual ∼257 kg urea (∼120 kg nitrogen) per hectare. Wheat plants arising from the seeds inoculated with the engineered Azotobacter strain, exhibited far superior overall performance, had much higher dry weight and nitrogen content and assimilated molecular 15 N much better. A nitrogen balance experiment also revealed much higher total nitrogen content. IAA production by the wild type and the engineered strain was about the same. Inoculation of the wheat seeds with A. chroococcum HKD15 did not adversely affect the microbial population in the field rhizosphere soil.IMPORTANCE Application of synthetic nitrogenous fertilizers is a standard agricultural practice to augment crop yield. Plants, however, utilize only a fraction of the applied fertilizers, while the unutilized fertilizers cause grave environmental problems. Wild type soil diazotrophic microrganisms cannot replace synthetic nitrogenous fertilizers, as these reduce atmospheric nitrogen

  5. Installation of the Tritium Purification System

    Energy Technology Data Exchange (ETDEWEB)

    Labik, G.; Golian, T.; Satkofsky, J.; Sichta, P.; Crook, D.; Dudek, L.; Coward, G.; Parsells, R. [Princeton Plasma Physics Lab., NJ (United States)


    This paper describes and details the design, the tasks, and the considerations for the mechanical and electrical installation of the TFTR Tritium Purification System (TPS) at the Princeton Plasma Physics Laboratory (PPPL). Canadian Fusion Fuels Technology Project (CFFTP) designed, fabricated, assembled and tested the Tritium Purification System in Ontario, Canada. After system tests were accepted by Princeton Plasma Physics Laboratory, the assembled components were disassembled into a set of subassemblies and were shipped to PPPL. The subassemblies were reassembled at PPPL and installed primarily in the Decon Facility. The original site selection was within the TFTR tritium processing area and that selection impacted the column design. The Decon Facility was later chosen to permit a better layout of equipment and improved access for installation personnel. Selection of the Decon Facility site resulted in longer line runs for most of the process streams including the tritium product line. The initial review of the proposed installation was conducted during September of 1994 and the System Integrated Test began during April 1995, subsequent to a successful Operational Readiness Assessment conducted during March of 1995.

  6. Online Oxide Contamination Measurement and Purification Demonstration (United States)

    Bradley, D. E.; Godfroy, T. J.; Webster, K. L.; Garber, A. E.; Polzin, K. A.; Childers, D. J.


    Liquid metal sodium-potassium (NaK) has advantageous thermodynamic properties indicating its use as a fission reactor coolant for a surface (lunar, martian) power system. A major area of concern for fission reactor cooling systems is system corrosion due to oxygen contaminants at the high operating temperatures experienced. A small-scale, approximately 4-L capacity, simulated fission reactor cooling system employing NaK as a coolant was fabricated and tested with the goal of demonstrating a noninvasive oxygen detection and purification system. In order to generate prototypical conditions in the simulated cooling system, several system components were designed, fabricated, and tested. These major components were a fully-sealed, magnetically-coupled mechanical NaK pump, a graphite element heated reservoir, a plugging indicator system, and a cold trap. All system components were successfully demonstrated at a maximum system flow rate of approximately 150 cc/s at temperatures up to 550 C. Coolant purification was accomplished using a cold trap before and after plugging operations which showed a relative reduction in oxygen content.

  7. Nylon wool purification alters the activation of T cells. (United States)

    Wohler, Jillian E; Barnum, Scott R


    Purification of lymphocytes, particularly T cells, is commonly performed using nylon wool. This enrichment method selectively retains B cells and some myeloid cells allowing a significantly more pure T cell population to flow through a nylon wool column. T cells purified in this fashion are assumed to be unaltered and functionally naïve, however some studies have suggested aberrant in vitro T cell responses after nylon wool treatment. We found that nylon wool purification significantly altered T cell proliferation, expression of activation markers and production of cytokines. Our results suggest that nylon wool treatment modifies T cell activation responses and that caution should be used when choosing this purification method.

  8. Effect of water purification process in radioactive content: analysis on small scale purification plants

    International Nuclear Information System (INIS)

    Lopez del Rio, H.; Quiroga S, J. C.; Davila R, J. I.; Mireles G, F.


    Water from small scale purification plants is a low cost alternative for consumers in comparison to the bottled commercial presentations. Because of its low cost per liter, the consumption of this product has increased in recent years, stimulating in turn the installation of purification systems for these small businesses. The purpose of this study was to estimate the efficiency of small scale purification systems located in the cities of Zacatecas and Guadalupe, Zacatecas, to reduce the radioactive content of water. It was measured the total alpha and beta activity in water samples of entry and exit to process, through the liquid scintillation technique. In general it was observed that the process is more efficient in removing alpha that beta activity. The fraction of total alpha activity removed varied between 27 and 100%, while between 0 and 77% of the total beta activity was removed by the analyzed plants. In all cases, the total radioactivity level was lower than the maximum permissible value settled by the official mexican standard for drinking water. (Author)

  9. Biomimetic affinity ligands for protein purification. (United States)

    Sousa, Isabel T; Taipa, M Angela


    The development of sophisticated molecular modeling software and new bioinformatic tools, as well as the emergence of data banks containing detailed information about a huge number of proteins, enabled the de novo intelligent design of synthetic affinity ligands. Such synthetic compounds can be tailored to mimic natural biological recognition motifs or to interact with key surface-exposed residues on target proteins and are designated as "biomimetic ligands." A well-established methodology for generating biomimetic or synthetic affinity ligands integrates rational design with combinatorial solid-phase synthesis and screening, using the triazine scaffold and analogues of amino acids side chains to create molecular diversity.Triazine-based synthetic ligands are nontoxic, low-cost, highly stable compounds that can replace advantageously natural biological ligands in the purification of proteins by affinity-based methodologies.

  10. Water purification using solar radiation in Nigeria

    International Nuclear Information System (INIS)

    Udounwa, A.E.; Osuji, R.U.


    In developing countries, lack of safe and reliable drinking water constitutes a major problem. Contaminated water is the major cause of most water borne diseases like diarrhoea. Disinfection of water is accomplished by a number of different physical - chemical treatments including boiling, application of chlorine and filtration techniques. Solar energy, which is universally available, can also be used effectively in this process, that is, to deactivate the micro-organisms present in this contaminated water thereby improving its microbiological quality. This treatment process is called solar water disinfection. This paper therefore appraises the extent to which research work has been done as regards purification of water using solar radiation in Nigeria vis-a-vis outside the country. It is hoped that it will serve as a wake-up-call for Nigerians especially those in remote areas with no treated pipe borne water supply. The problems and prospects of this technology as well as the policy implications are presented. (author)

  11. Radioligand purification prior to routine receptor assays

    International Nuclear Information System (INIS)

    Le Goff, J.-M.; Berthois, Y.; Martin, P.-M.


    The need to repurify the commercially available radioligands [ 3 H]estradiol and [ 3 H]testosterone before use in routine assays was investigated. Storage of these products for 2 months after delivery led to appreciable degradation of [ 3 H]estradiol compared to [ 3 H]testosterone. Unexpectedly, TLC and even HPLC procedures were ineffective in completely restoring the purity of [ 3 H]-estradiol and the unremoved polar products induced important variations in our estrogen receptor assays. An increase in non-specific binding and a concomitant decrease in total binding were observed resulting in an underestimation of specific binding sites and of the affinity constant. In some cases Scatchard analysis was not possible. The authors therefore strongly recommend the repurification of low-stability radioligands and propose an economic time-saving procedure for the purification of [ 3 H]estradiol by solvent differential partition which requires no high-cost investment in apparatus. (author)

  12. Purification of equine Gc-globulin

    DEFF Research Database (Denmark)

    Houen, Gunnar; Pihl, Tina Holberg; Andersen, Pia Haubro

    Objectives With the aim of producing antibodies for an equine Group specific component (Gc)-globulin assay, the protein was purified from normal equine plasma. Methods Equine Gc-globulin was purified from healthy horse plasma using ion exchange chromatography (Q-Sepharose, CM......-Sepharose) and preparative PAGE. Results Equine Gc-globulin has successfully been purified from healthy horse plasma and rabbits and mice are being immunized to produce specific antibodies. Conclusions Purification of equine Gc-globulin and the production of specific antibodies will make it possible to develop an assay...... to be a sensitive marker of acute tissue injury and fatal outcome in humans. Patients with a low plasma concentration of Gc-globulin due to severe tissue injury might potentially benefit from infusions with purified Gc-globulin [1]. With an equine Gc-globulin assay, future studies will investigate the concentration...

  13. Submersible purification system for radioactive water (United States)

    Abbott, Michael L.; Lewis, Donald R.


    A portable, submersible water purification system for use in a pool of water containing radioactive contamination includes a prefilter for filtering particulates from the water. A resin bed is then provided for removal of remaining dissolved, particulate, organic, and colloidal impurities from the prefiltered water. A sterilizer then sterilizes the water. The prefilter and resin bed are suitably contained and are submerged in the pool. The sterilizer is water tight and located at the surface of the pool. The water is circulated from the pool through the prefilter, resin bed, and sterilizer by suitable pump or the like. In the preferred embodiment, the resin bed is contained within a tank which stands on the bottom of the pool and to which a base mounting the prefilter and pump is attached. An inlet for the pump is provided adjacent the bottom of the pool, while the sterilizer and outlet for the system is located adjacent the top of the pool.

  14. Purification of biomaterials by phase partitioning (United States)

    Harris, J. M.


    A technique which is particularly suited to microgravity environments and which is potentially more powerful than electrophoresis is phase partitioning. Phase partitioning is purification by partitioning between the two immiscible aqueous layers formed by solution of the polymers poly(ethylene glycol) and dextran in water. This technique proved to be very useful for separations in one-g but is limited for cells because the cells are more dense than the phase solutions thus tend to sediment to the bottom of the container before reaching equilibrium with the preferred phase. There are three phases to work in this area: synthesis of new polymers for affinity phase partitioning; development of automated apparatus for ground-based separations; and design of apparatus for performing simple phase partitioning space experiments, including examination of mechanisms for separating phases in the absence of gravity.

  15. Nanocellulose-Based Materials for Water Purification. (United States)

    Voisin, Hugo; Bergström, Lennart; Liu, Peng; Mathew, Aji P


    Nanocellulose is a renewable material that combines a high surface area with high strength, chemical inertness, and versatile surface chemistry. In this review, we will briefly describe how nanocellulose is produced, and present-in particular, how nanocellulose and its surface modified versions affects the adsorption behavior of important water pollutants, e.g., heavy metal species, dyes, microbes, and organic molecules. The processing of nanocellulose-based membranes and filters for water purification will be described in detail, and the uptake capacity, selectivity, and removal efficiency will also be discussed. The processing and performance of nanocellulose-based membranes, which combine a high removal efficiency with anti-fouling properties, will be highlighted.

  16. Nanocellulose-Based Materials for Water Purification

    Directory of Open Access Journals (Sweden)

    Hugo Voisin


    Full Text Available Nanocellulose is a renewable material that combines a high surface area with high strength, chemical inertness, and versatile surface chemistry. In this review, we will briefly describe how nanocellulose is produced, and present—in particular, how nanocellulose and its surface modified versions affects the adsorption behavior of important water pollutants, e.g., heavy metal species, dyes, microbes, and organic molecules. The processing of nanocellulose-based membranes and filters for water purification will be described in detail, and the uptake capacity, selectivity, and removal efficiency will also be discussed. The processing and performance of nanocellulose-based membranes, which combine a high removal efficiency with anti-fouling properties, will be highlighted.

  17. Purification of tritium-free water

    International Nuclear Information System (INIS)

    Hussain, S.D.


    Ground water which has been out of contact with the atmosphere for a long time as compared to the half life of tritium (12.43 years) does not contain any measureable amount of tritium. Such water is called tritium-free water. It may contain dissolved and suspended impurities and has to be purified before it can be used for the preparation of blanks and standards required in the routine measurement of low level tritium in water samples. The purification of tritium-free water by distillation in a closed system has been described. The quality of processed tritium-free water was precisely checked at International Atomic Energy Agency (IAEA) Vienna and found satisfactory. (authors)

  18. Purification and characterization of amine oxidase from Vigna ...

    African Journals Online (AJOL)



    Sepharose chromatography to 544 purification folds with .... mungo L. seeds were obtained from local market. Horseradish peroxidase ... volume of 50 mM potassium phosphate buffer (pH 8.0), dialyzed 3 times against the same ...

  19. Production and purification of polyclonal anti-hamster ...

    African Journals Online (AJOL)

    . ... IgG showed high titer and high specificity in the designed ELISA. Purified antibody and its conjugation with HRP are used in research and diagnosis of hamster disease. Key words: Production, purification, hamster immunoglobulins.

  20. Radiation-adsorption purification of effluents containing pesticides

    International Nuclear Information System (INIS)

    Brusentseva, S.A.; Shubin, V.N.; Nikonorova, G.K.; Zorin, D.M.; Sosnovskaya, A.A.; Petryaev, E.P.; Vlasova, V.I.; Edimicheva, I.P.; Subbotina, N.N.; Belorusskij Gosudarstvennyj Univ., Minsk)


    The radiation-adsorption purification is one of the new direction in the radiation purification of natural wastes and effluents containing pesticides. This method combines the conventional adsorption purification with radiation treatment of the sorbent, and the result the protection time of the sorbent increases due to the radiation regeneration of carbon. In present work the method was used for purification of effluents from pesticides, such as 4,4'Dichlorodiphenyltrichloroethane /DDT/, 1,2,3,4,5,6-hexachlorocyclohexane /HCCH/, dimethyl 2,2-dichlorovinylphosphate /DDVF/ and petroleum products (a mixture of kerosene and xylene in ratio 7:1). Such effluents are formed at factories producing an insecticide aerosol 'Prime-71'. Three investigations were carried out on model with a solution similar composition to industrial effluents. (author)

  1. Microbiological and technical aspects of anaerobic waste water purification

    International Nuclear Information System (INIS)

    Aivasidis, A.


    Anaerobic waste water purification is likely to be another example of how innovations can result from the joint use of biological and technical concepts. No matter how far the optimization of oxygen input with aerobic waste water purification advances it will still be the less a real competitor for anaerobic techniques the more polluted the waste water is. The principle of carrier fixation to avoid their washing out, too, has often been observed in nature with sessile microorganisms. With highly polluted water, anaerobic purification does not only work at no expenditure of energy but it can also make excess energy available for use in other processes. Another important argument for anaerobic methods of waste water purification is probably the clearly reduced production of excess sludge. (orig.) [de

  2. Optimized conditions for high-level solubilization and purification of ...

    African Journals Online (AJOL)

    Optimized conditions for high-level solubilization and purification of recombinant camel growth hormone in Escherichia coli. MAN Saqib, M Fatima, IN Awan, MI Shahzad, SKA Rizvi, M Mukhtar, A Khanum ...

  3. Purification and characterization of a peroxidase present in ...

    African Journals Online (AJOL)

    Purification and characterization of a peroxidase present in xilopodium exsudates of umbu plants (Spondias tuberosa A.) M dos Santos Teixeira Pinto, JM Ribeiro, FP de Araujo, NF de Melo, KVS Fernandes ...

  4. Purification and characterization of three laccase isozymes from the ...

    African Journals Online (AJOL)

    Purification and characterization of three laccase isozymes from the white rot fungus Trametes sp. HS-03. Weiyun Guo, Zhaoyang Yao, Chenyan Zhou, Duan Li, Hongli Chen, Qiang Shao, Zongyi Li, Huigen Feng ...

  5. Security proof of quantum cryptography based entirely on entanglement purification

    International Nuclear Information System (INIS)

    Aschauer, Hans; Briegel, Hans J.


    We give a proof that entanglement purification, even with noisy apparatus, is sufficient to disentangle an eavesdropper (Eve) from the communication channel. In the security regime, the purification process factorizes the overall initial state into a tensor-product state of Alice and Bob, on one side, and Eve on the other side, thus establishing a completely private, albeit noisy, quantum communication channel between Alice and Bob. The security regime is found to coincide for all practical purposes with the purification regime of a two-way recurrence protocol. This makes two-way entanglement purification protocols, which constitute an important element in the quantum repeater, an efficient tool for secure long-distance quantum cryptography

  6. Optimized conditions for high-level solubilization and purification of ...

    African Journals Online (AJOL)

    NH Computers


    Oct 10, 2011 ... Key words: Recombinant growth hormone, somatotropin, cloning, expression, inclusion bodies, solubilization, purification, bioactivity. ... Lactating animals growth hormones (GH) have been exploited through biotechnology to increase ..... Kostyo JL, Isaksson O (1977). Growth hormone and the regulation of.

  7. Purification, composition analysis and antioxidant activity of different ...

    African Journals Online (AJOL)

    Purification, composition analysis and antioxidant activity of different polysaccharides from the fruiting bodies of Pholiota adiposa. Yongxin Nie, Hongxia Jiang, Yanyou Su, Changxiang Zhu, Jiarui Li, Fujiang Wen ...

  8. Purification and biochemical characterization of a novel glutathione ...

    African Journals Online (AJOL)



    dinitrobenzene as a substrate. ..... purification of GST enzyme from other insect species, which were from 3 to 26% in Hyphantria .... Glutathione S-transferases from the larval gut of the silkworm. Bombyx mori: cDNA cloning, gene ...

  9. Feasibility Study on Manufacturing Lightweight Aggregates from Water Purification Sludge (United States)

    Peng, Ching-Fang; Chen, How-Ji


    This study mainly discussed the feasibility of manufacturing lightweight aggregates from water purification sludge in Taiwan. They were analysed for the physical and chemical composition before the sintering test for lightweight aggregates in a laboratory. Then the physical and mechanical properties of the synthesized aggregates were assessed. The result showed that the chemical composition of sludge in the water purification plants was within the appropriate range for manufacturing lightweight aggregate as proposed in the literature. The sintering test demonstrated that the particle density of aggregates from the ten types of water purification sludge were mostly less than 1.8 g/cm3. In addition, the dry unit weight, the organic impurity, the ignition loss, and other characteristics of synthesized aggregates met the requirement of CNS standards, while its water absorption and crushing strength also fulfilled the general commercial specifications. Therefore, reclamation of water purification sludge for production of lightweight aggregate is indeed feasible.

  10. Purification and Characterization of Lipase from Aspergillus flavus ...

    African Journals Online (AJOL)

    Lipase from Aspergillus flavus was purified in a single step purification using MnFeO4 magnetic nano particles to achieve a 20.53- fold purification with specific activity of 11.29 U/mg and a 59% recovery yield. SDS-PAGE of lipase showed a single pure band with corresponding molecular weight of 35 kDa. The optimal ...

  11. Specialists' meeting on fast reactor cover gas purification

    International Nuclear Information System (INIS)


    The tentative agenda was adopted by the participants without comment and was followed throughout the meeting. The following topics were discussed at the subsequent sessions of the meeting on 'Fast Reactor Cover Gas Purification': National Position Papers; Impurities: Sources and Measurement; Cover Gas Purification Techniques; Sodium Aerosol Trapping; Radiological Considerations. Based on the papers presented and the discussions following, session summaries and conclusions were prepared and are included in this report

  12. Purification of human platelet-derived growth factor

    International Nuclear Information System (INIS)

    Raines, E.W.; Ross, R.


    The paper describes a method for purification of human platelet-derived growth factor (PDGF) from outdated platelet-rich plasma (PRP) using commonly available laboratory reagents and yielding a mitogen purified 800,000-fold over the starting material. [ 3 H]thymidine incorporation into DNA of cultured cells responsive to PDGF represents the most readily available method to follow its purification and define the biological activity of a purified preparation. Other assays to quantitate PDGF include radioreceptor assay and radioimmunoassay

  13. Recommended methods for purification of solvents and tests for impurities

    CERN Document Server

    Coetzee, J F


    Recommended Methods for Purification of Solvents and Tests for Impurities is a compilation of recommended procedures for purification of solvents and tests for solvent impurities. Ten solvents are covered: acetonitrile, sulfolane, propylene carbonate, dimethyl sulfoxide, dimethylformamide, hexamethylphosphoramide, pyridine, ethylenediamine, N-methylacetamide, and N-methylpropionamide. This book is comprised of 12 chapters and opens with an introduction to general aspects of impurity effects. The rationale for the selection of solvent is explained, and the relative reactivities of solutes in di

  14. A simple method for purification of herpesvirus DNA

    DEFF Research Database (Denmark)

    Christensen, Laurids Siig; Normann, Preben


    A rapid and reliable method for purification of herpesvirus DNA from cell cultures is described. The method is based on the isolation of virus particles and/or nucleocapsids by differential centrifugation and exploits the solubilizing and denaturing capabilities of cesium trifluoroacetate during ...... isopycnic centrifugation, so that phenol/chloroform extractions can be omitted. The method was used for the purification of DNA from several members of the Alfaherpesvirinae subfamily....

  15. Development of fuzzy logic algorithm for water purification plant




    This paper propose the design of FLC algorithm for industrial application such application is water purification plant. In the water purification plant raw water or ground water is promptly purified by injecting chemical at rates related to water quality. The feed of chemical rates judged and determined by the skilled operator. Yagishita et al.[1] structured a system based on fuzzy logic so that the feed rate of the coagulant can be judged automatically without any skilled operator. We perfor...

  16. Purification of cerium, neodymium and gadolinium for low background experiments

    Directory of Open Access Journals (Sweden)

    Boiko R.S.


    Full Text Available Cerium, neodymium and gadolinium contain double beta active isotopes. The most interesting are 150Nd and 160Gd (promising for 0ν2β search, 136Ce (2β+ candidate with one of the highest Q2β. The main problem of compounds containing lanthanide elements is their high radioactive contamination by uranium, radium, actinium and thorium. The new generation 2β experiments require development of methods for a deep purification of lanthanides from the radioactive elements. A combination of physical and chemical methods was applied to purify cerium, neodymium and gadolinium. Liquid-liquid extraction technique was used to remove traces of Th and U from neodymium, gadolinium and for purification of cerium from Th, U, Ra and K. Co-precipitation and recrystallization methods were utilized for further reduction of the impurities. The radioactive contamination of the samples before and after the purification was tested by using ultra-low-background HPGe gamma spectrometry. As a result of the purification procedure the radioactive contamination of gadolinium oxide (a similar purification efficiency was reached also with cerium and neodymium oxides was decreased from 0.12 Bq/kg to 0.007 Bq/kg in 228Th, from 0.04 Bq/kg to <0.006 Bq/kg in 226Ra, and from 0.9 Bq/kg to 0.04 Bq/kg in 40K. The purification methods are much less efficient for chemically very similar radioactive elements like actinium, lanthanum and lutetium.

  17. Membrane Purification Cell for Aluminum Recycling

    Energy Technology Data Exchange (ETDEWEB)

    David DeYoung; James Wiswall; Cong Wang


    Recycling mixed aluminum scrap usually requires adding primary aluminum to the scrap stream as a diluent to reduce the concentration of non-aluminum constituents used in aluminum alloys. Since primary aluminum production requires approximately 10 times more energy than melting scrap, the bulk of the energy and carbon dioxide emissions for recycling are associated with using primary aluminum as a diluent. Eliminating the need for using primary aluminum as a diluent would dramatically reduce energy requirements, decrease carbon dioxide emissions, and increase scrap utilization in recycling. Electrorefining can be used to extract pure aluminum from mixed scrap. Some example applications include producing primary grade aluminum from specific scrap streams such as consumer packaging and mixed alloy saw chips, and recycling multi-alloy products such as brazing sheet. Electrorefining can also be used to extract valuable alloying elements such as Li from Al-Li mixed scrap. This project was aimed at developing an electrorefining process for purifying aluminum to reduce energy consumption and emissions by 75% compared to conventional technology. An electrolytic molten aluminum purification process, utilizing a horizontal membrane cell anode, was designed, constructed, operated and validated. The electrorefining technology could also be used to produce ultra-high purity aluminum for advanced materials applications. The technical objectives for this project were to: - Validate the membrane cell concept with a lab-scale electrorefining cell; - Determine if previously identified voltage increase issue for chloride electrolytes holds for a fluoride-based electrolyte system; - Assess the probability that voltage change issues can be solved; and - Conduct a market and economic analysis to assess commercial feasibility. The process was tested using three different binary alloy compositions (Al-2.0 wt.% Cu, Al-4.7 wt.% Si, Al-0.6 wt.% Fe) and a brazing sheet scrap composition (Al-2

  18. Purification of equine Gc-globulin

    DEFF Research Database (Denmark)

    Houen, Gunnar; Pihl, Tina Holberg; Andersen, Pia Haubro

    Objectives With the aim of producing antibodies for an equine Group specific component (Gc)-globulin assay, the protein was purified from normal equine plasma. Methods Equine Gc-globulin was purified from healthy horse plasma using ion exchange chromatography (Q-Sepharose, CM-Sepharose) and prepa......Objectives With the aim of producing antibodies for an equine Group specific component (Gc)-globulin assay, the protein was purified from normal equine plasma. Methods Equine Gc-globulin was purified from healthy horse plasma using ion exchange chromatography (Q-Sepharose, CM......-Sepharose) and preparative PAGE. Results Equine Gc-globulin has successfully been purified from healthy horse plasma and rabbits and mice are being immunized to produce specific antibodies. Conclusions Purification of equine Gc-globulin and the production of specific antibodies will make it possible to develop an assay...... for measuring Gc-globulin in horses. Studies in rodents and humans have shown that Gc-globulin is a multifunctional acute phase plasma protein, which removes actin from the blood by binding it and facilitating its clearance from the circulation by the liver. As such, Gc-globulin prevents hyper coagulation...

  19. Commercial Charcoal Characterisation For Water Purification

    International Nuclear Information System (INIS)

    Saryati; Sumardjo; Sutisna; Handayani, Ari; Suprapti, Siti


    In order to provide a drinking water purification substance, has been studied the charcoal characterisation that based on a porous profile and an adsorption properties of the charcoal. There were using the commercial charcoal like wood charcoals, coconut shell charcoals and activated charcoals. The porous profile was studied by using an electron microscope SEM-EDX and the adsorption properties was studied by using the water sample simulation that contains several metal ions. The concentration of all ions was ten times greater that the maximum ions concentration that permissible in the drinking water. From the grain surface microscopic analysis was shown that the pore structure of the wood charcoal was more regular than the coconut shell charcoal. Mean while the activated charcoal has pore more than wood and coconut shell charcoal. Grains size was not an adsorption parameter. The absorptivitas charcoal was affected by pH solution, but this effect was not linear proportion. There are no significant deference in the adsorptivitas among the tree charcoals that has been studied for Al 3 + , Cr 3+ , Ag 1 +, and Pb 2+ ions the adsorption was large enough (> 60%), for Mn 2+ , Fe 3+ , Se 4+ , Cd 2+ and Ba 2+ ions was 20%-60% dan for Mg 2+ , Na 1+ , Ca 2+ , and Zn 2+ ions was less than 20 %. Generally the wood and coconut shell charcoal absorptivity in the pH 4 solutions was lower than in the pH 5-7 solutions

  20. Isolation and Purification of Biotechnological Products (United States)

    Hubbuch, Jürgen; Kula, Maria-Regina


    The production of modern pharma proteins is one of the most rapid growing fields in biotechnology. The overall development and production is a complex task ranging from strain development and cultivation to the purification and formulation of the drug. Downstream processing, however, still accounts for the major part of production costs. This is mainly due to the high demands on purity and thus safety of the final product and results in processes with a sequence of typically more than 10 unit operations. Consequently, even if each process step would operate at near optimal yield, a very significant amount of product would be lost. The majority of unit operations applied in downstream processing have a long history in the field of chemical and process engineering; nevertheless, mathematical descriptions of the respective processes and the economical large-scale production of modern pharmaceutical products are hampered by the complexity of the biological feedstock, especially the high molecular weight and limited stability of proteins. In order to develop new operational steps as well as a successful overall process, it is thus a necessary prerequisite to develop a deeper understanding of the thermodynamics and physics behind the applied processes as well as the implications for the product.

  1. The Purification of a Blood Group A Glycoprotein: An Affinity Chromatography Experiment. (United States)

    Estelrich, J.; Pouplana, R.


    Describes a purification process through affinity chromatography necessary to obtain specific blood group glycoproteins from erythrocytic membranes. Discusses the preparation of erythrocytic membranes, extraction of glycoprotein from membranes, affinity chromatography purification, determination of glycoproteins, and results. (CW)


    NARCIS (Netherlands)


    When membrane proteins are solubilized and subjected to purification procedures, the loss of lipids surrounding the protein often results in irreversible inactivation. We describe a procedure for the immunoaffinity purification of the membrane protein UDP-glucuronosyltransferase from human liver.

  3. Argon Collection And Purification For Proliferation Detection

    Energy Technology Data Exchange (ETDEWEB)

    Achey, R. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Hunter, D. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)


    In order to determine whether a seismic event was a declared/undeclared underground nuclear weapon test, environmental samples must be taken and analyzed for signatures that are unique to a nuclear explosion. These signatures are either particles or gases. Particle samples are routinely taken and analyzed under the Comprehensive Nuclear-Test-Ban Treaty Organization (CTBTO) verification regime as well as by individual countries. Gas samples are analyzed for signature gases, especially radioactive xenon. Underground nuclear tests also produce radioactive argon, but that signature is not well monitored. A radioactive argon signature, along with other signatures, can more conclusively determine whether an event was a nuclear test. This project has developed capabilities for collecting and purifying argon samples for ultra-low-background proportional counting. SRNL has developed a continuous gas enrichment system that produces an output stream containing 97% argon from whole air using adsorbent separation technology (the flow diagram for the system is shown in the figure). The vacuum swing adsorption (VSA) enrichment system is easily scalable to produce ten liters or more of 97% argon within twelve hours. A gas chromatographic separation using a column of modified hydrogen mordenite molecular sieve has been developed that can further purify the sample to better than 99% purity after separation from the helium carrier gas. The combination of these concentration and purification systems has the capability of being used for a field-deployable system for collecting argon samples suitable for ultra-low-background proportional counting for detecting nuclear detonations under the On-Site Inspection program of the CTBTO verification regime. The technology also has applications for the bulk argon separation from air for industrial purposes such as the semi-conductor industry.

  4. Purification and characterization of riproximin from Ximenia americana fruit kernels. (United States)

    Bayer, Helene; Ey, Noreen; Wattenberg, Andreas; Voss, Cristina; Berger, Martin R


    Highly pure riproximin was isolated from the fruit kernels of Ximenia americana, a defined, seasonally available and potentially unlimited herbal source. The newly established purification procedure included an initial aqueous extraction, removal of lipids with chloroform and subsequent chromatographic purification steps on a strong anion exchange resin and lactosyl-Sepharose. Consistent purity and stable biological properties were shown over several purification batches. The purified, kernel-derived riproximin was characterized in comparison to the African plant material riproximin and revealed highly similar biochemical and biological properties but differences in the electrophoresis pattern and mass spectrometry peptide profile. Our results suggest that although the purified fruit kernel riproximin consists of a mixture of closely related isoforms, it provides a reliable basis for further research and development of this type II ribosome inactivating protein (RIP). Copyright © 2011 Elsevier Inc. All rights reserved.

  5. The Status of KamLAND After Purification (United States)

    Grant, Christopher


    KamLAND is a 1-kton liquid scintillation detector located in the Kamioka underground laboratory, in Japan. KamLAND has provided a precision measurement of δm^221 using reactor anti-neutrinos, and yielded first observational evidence of geologically produced anti-neutrinos. Since April of 2007, the collaboration has been working on the purification of the detector with the goal of observing 862 keV, ^7Be solar neutrinos. Two purification campaigns have concluded, with a total of 5.4 ktons of scintillator circulated through a distillation and nitrogen purge system. The results of purification and the overall background reduction factors will be presented, along with an update of the ^7Be solar neutrino analysis. )

  6. Recent Advances in Nanoporous Membranes for Water Purification

    Directory of Open Access Journals (Sweden)

    Zhuqing Wang


    Full Text Available Nanoporous materials exhibit wide applications in the fields of electrocatalysis, nanodevice fabrication, energy, and environmental science, as well as analytical science. In this review, we present a summary of recent studies on nanoporous membranes for water purification application. The types and fabrication strategies of various nanoporous membranes are first introduced, and then the fabricated nanoporous membranes for removing various water pollutants, such as salt, metallic ions, anions, nanoparticles, organic chemicals, and biological substrates, are demonstrated and discussed. This work will be valuable for readers to understand the design and fabrication of various nanoporous membranes, and their potential purification mechanisms towards different water pollutants. In addition, it will be helpful for developing new nanoporous materials for quick, economic, and high-performance water purification.

  7. Study of air purification in the production of radioactive compounds

    International Nuclear Information System (INIS)

    Fradin, J.; Desroches, J.


    As the fabrication of radio-elements takes place in almost airtight enclosures in which a frequent air renewal is required, and while taking the purification rate into account (1.000 to 5.000 m 3 /h), the authors report the study of wet purification in conjunction with dry purification through paper filters, in order to capture dusts. An apparatus has been implemented which allows high gas flow rates. A radioactive aerosol has been introduced in this apparatus and its efficiency has been measured by different means. The authors describe the instrumentation (column, aerosol generator), operation, rate adjustment, and losses. Aerosols of manganese and sodium have been used. Their particle granulometry has been determined. The authors report several types of tests [fr

  8. Bromelain: an overview of industrial application and purification strategies. (United States)

    Arshad, Zatul Iffah Mohd; Amid, Azura; Yusof, Faridah; Jaswir, Irwandi; Ahmad, Kausar; Loke, Show Pau


    This review highlights the use of bromelain in various applications with up-to-date literature on the purification of bromelain from pineapple fruit and waste such as peel, core, crown, and leaves. Bromelain, a cysteine protease, has been exploited commercially in many applications in the food, beverage, tenderization, cosmetic, pharmaceutical, and textile industries. Researchers worldwide have been directing their interest to purification strategies by applying conventional and modern approaches, such as manipulating the pH, affinity, hydrophobicity, and temperature conditions in accord with the unique properties of bromelain. The amount of downstream processing will depend on its intended application in industries. The breakthrough of recombinant DNA technology has facilitated the large-scale production and purification of recombinant bromelain for novel applications in the future.

  9. Purification of thyrotropin from human hypophysis: preliminary preparation

    International Nuclear Information System (INIS)

    Borghi, V.C.; Lin, L.H.; Bartolini, P.


    The adequacy of stored crude preparations for isolation of human tyrotropin (TSH) was evaluated according to Ross et al from a side fraction obtained during the purification of growth hormone from frozen pituitaries (SOMATORMON). Six crude TSH preparations were stored at - 20 0 C during several years for further purification. One of these preparations was purified by sucessive chromatographies on Sephadex G-100, hydroxylapatite and SP-Sephadex C50. The TSH content present in the chromatographic fractions and in the pools was assayed by specific radioimmunoassay developed at our laboratory. The protein determination of the fractions and pools was performed by absorbance at 280 nm and by the method of Lowry at al, respectively. The TSH activity increased eight times during the purification and the TSH purified had a radioimmunological potency around half that de scribed by Roos at al. The results suggest the fitness of long time stored preparations in the attainment of pure TSH. (author) [pt

  10. The status of international purification process of uranium

    International Nuclear Information System (INIS)

    Huang Lunguan; Zhuang Haixing; Zhuo Jianwei; Jiang Ping; Niu Yuqing


    The information of international purification process of uranium since 1976 is retrieved by both computer and handwork. The uranium purification processes and parameters in the United States, England, France, Canada, India, Korea, Australia, Pakistan, Brazil, Spain, Philippines, Japan, Argentina and China are summarized. The review show that, in addition to amine extraction in Japan and Argentina, TBP extraction was normally used in the rest countries. In order to reduce the volume of equipment and consumption of reagents, the feed solution with high uranium content (more than 300 g/L uranium) was adopted by almost all countries. During scrubbing of loaded organic phase, the methods using partly returned product liquor were studied in some countries and columns were mainly used as extraction equipment so as to improve the purification efficiency. In some countries, pulp extraction was studied to overcome the difficulties in the process of filtering

  11. Purification of crime scene DNA extracts using centrifugal filter devices. (United States)

    Norén, Lina; Hedell, Ronny; Ansell, Ricky; Hedman, Johannes


    The success of forensic DNA analysis is limited by the size, quality and purity of biological evidence found at crime scenes. Sample impurities can inhibit PCR, resulting in partial or negative DNA profiles. Various DNA purification methods are applied to remove impurities, for example, employing centrifugal filter devices. However, irrespective of method, DNA purification leads to DNA loss. Here we evaluate the filter devices Amicon Ultra 30 K and Microsep 30 K with respect to recovery rate and general performance for various types of PCR-inhibitory crime scene samples. Recovery rates for DNA purification using Amicon Ultra 30 K and Microsep 30 K were gathered using quantitative PCR. Mock crime scene DNA extracts were analyzed using quantitative PCR and short tandem repeat (STR) profiling to test the general performance and inhibitor-removal properties of the two filter devices. Additionally, the outcome of long-term routine casework DNA analysis applying each of the devices was evaluated. Applying Microsep 30 K, 14 to 32% of the input DNA was recovered, whereas Amicon Ultra 30 K retained 62 to 70% of the DNA. The improved purity following filter purification counteracted some of this DNA loss, leading to slightly increased electropherogram peak heights for blood on denim (Amicon Ultra 30 K and Microsep 30 K) and saliva on envelope (Amicon Ultra 30 K). Comparing Amicon Ultra 30 K and Microsep 30 K for purification of DNA extracts from mock crime scene samples, the former generated significantly higher peak heights for rape case samples (P-values crime scene samples and for consistency between different PCR-based analysis systems, such as quantification and STR analysis. In order to maximize the possibility to obtain complete STR DNA profiles and to create an efficient workflow, the level of DNA purification applied should be correlated to the inhibitor-tolerance of the STR analysis system used.

  12. Enhancement of Chlorella vulgaris growth and bioremediation ability of aquarium wastewater using diazotrophs. (United States)

    Ali, Sayeda Mohammed; Nasr, Hoda Shafeek; Abbas, Wafaa Tawfik


    Treatment of aquarium wastewater represents an important process to clean and recycle wastewater to be safely returned to the environment, used for cultivation or to minimize the multiple renewal of water. Chlorella vulgaris was an important freshwater microalgae which used in wastewater treatment, and increasing its potential of treatment can be achieved with existence of N2-fixing bacteria. Co-culturing of Chlorella vulgaris with the diazotrophs, Azospirillum brasilense or Azotobacter chroococcum in three different media; aquarium wastewater (AWW), sterile enriched natural aquarium wastewater (GPM) and synthetic wastewater media (SWW) were studied. Biomass yield of the microalgae was estimated by determination of chlorophylls (a and b), total carotenoid and the dry weight of C. vulgaris. Also determination of ammonia, nitrite, phosphate and nitrate in the culture were done. The presence of diazotrophs significantly increased the biomass of C. vulgaris by increasing its microalgae pigments (chlorophylls a and b, and total carotenoids). The highest pigments percentage was reported due to addition of A. brasilense to C. vulgaris (18.3-133.5%) compared to A. chroococcum (23.9-56.9%). As well as increased dry weight from 12 to 50%. There was also improved removal of nitrate, nitrite, ammonia and phosphate; where, the highest removal percentage was reported due to addition of A. chroococcum to C. vulgaris (0.0-52%) compared to A. brasilense (0.6-16.4%). A. brasilense and A. chroococcum can support C. vulgaris biomass production and bioremediation activity in the aquarium to minimize the periodical water renewal.

  13. High quality protein microarray using in situ protein purification

    Directory of Open Access Journals (Sweden)

    Fleischmann Robert D


    Full Text Available Abstract Background In the postgenomic era, high throughput protein expression and protein microarray technologies have progressed markedly permitting screening of therapeutic reagents and discovery of novel protein functions. Hexa-histidine is one of the most commonly used fusion tags for protein expression due to its small size and convenient purification via immobilized metal ion affinity chromatography (IMAC. This purification process has been adapted to the protein microarray format, but the quality of in situ His-tagged protein purification on slides has not been systematically evaluated. We established methods to determine the level of purification of such proteins on metal chelate-modified slide surfaces. Optimized in situ purification of His-tagged recombinant proteins has the potential to become the new gold standard for cost-effective generation of high-quality and high-density protein microarrays. Results Two slide surfaces were examined, chelated Cu2+ slides suspended on a polyethylene glycol (PEG coating and chelated Ni2+ slides immobilized on a support without PEG coating. Using PEG-coated chelated Cu2+ slides, consistently higher purities of recombinant proteins were measured. An optimized wash buffer (PBST composed of 10 mM phosphate buffer, 2.7 mM KCl, 140 mM NaCl and 0.05% Tween 20, pH 7.4, further improved protein purity levels. Using Escherichia coli cell lysates expressing 90 recombinant Streptococcus pneumoniae proteins, 73 proteins were successfully immobilized, and 66 proteins were in situ purified with greater than 90% purity. We identified several antigens among the in situ-purified proteins via assays with anti-S. pneumoniae rabbit antibodies and a human patient antiserum, as a demonstration project of large scale microarray-based immunoproteomics profiling. The methodology is compatible with higher throughput formats of in vivo protein expression, eliminates the need for resin-based purification and circumvents

  14. Purification of a synthetic pterocarpanquinone by countercurrent chromatography

    International Nuclear Information System (INIS)

    Costa, Fernanda das Neves; Silva, Alcides Jose M. da; Domingos, Jorge L. de Oliveira; Costa, Paulo Roberto R.; Leitao, Gilda G.; Daher Netto, Chaquip


    Countercurrent chromatography (CCC) was employed as a useful, fast and economic alternative to conventional chromatography techniques for the purification of a synthetic pterocarpanquinone, LQB-118. The separation was performed in a two-step CCC with the solvent system hexanechloroform- methanol-water 2:1.5:5:2 in both steps. Traditional purification of these reaction products by silica gel column chromatography demanded a large amount of solvent and time, besides allowing the irreversible adsorption of the compound in the column. The use of 1 H NMR for the calculation of KD of target compound is proposed as an alternative for HPLC measurements. (author)

  15. Sandia Sodium Purification Loop (SNAPL) description and operations manual

    International Nuclear Information System (INIS)

    Acton, R.U.; Weatherbee, R.L.; Smith, L.A.; Mastin, F.L.; Nowotny, K.E.


    Sandia's Sodium Purification Loop was constructed to purify sodium for fast reactor safety experiments. An oxide impurity of less than 10 parts per million is required by these in-pile experiments. Commercial, reactor grade sodium is purchased in 180 kg drums. The sodium is melted and transferred into the unit. The unit is of a loop design and purification is accomplished by ''cold trapping.'' Sodium purified in this loop has been chemically analysed at one part per million oxygen by weight. 5 refs., 22 figs., 7 tabs

  16. Extraction and purification of plutonium by a tertiary amine

    International Nuclear Information System (INIS)

    Trentinian, M. de; Chesne, A.; Commissariat a l'Energie Atomique, Saclay


    Trilaurylamine diluted with a paraffinic solvent (dodecane) was studied as part of the research dealing with the separation and purification of plutonium. The physical properties (solubility of nitrates in the amine as a function of temperature) and the resistance to radiations of this substance were examined. The extraction characteristics of nitric solutions of plutonium, uranium and certain fission products are given as a function of the following factors: concentration of the various ions in solution, valency states. A method of plutonium purification based on these results is presented. (author) [fr

  17. Kinetics of nitrogenase from Azotobacter vinelandii

    NARCIS (Netherlands)

    Duyvis, M.G.


    Nitrogenase has been the subject of many investigations since the early 1960's. The catalytic mechanism of nitrogenase is unique because it couples the transfer of electrons with the hydrolysis of MgATP. The details of the mechanism are still to be revealed. The work described in this

  18. Purification of highly chlorinated dioxins degrading enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Ishii, K.; Furuichi, T.; Koike, K.; Kuboshima, M. [Hokkaido Univ. (Japan). Division of Environment Resource Engineering, Graduate School of Engineering


    Soil contamination caused by dioxins in and around sites of incinerators for municipal solid waste (MSW) is a concern in Japan. For example, scattering wastewater from a wet gas scrubber at an MSW incinerator facility in Nose, Osaka caused soil and surface water contamination. The concentration of dioxins in the soil was about 8,000 pg-TEQ/g. Other contamination sites include soils on which fly ash has been placed directly or improperly stored and landfill sites that have received bottom and fly ash over a long period. Some countermeasures are required immediately at these dioxins-contaminated sites. We have previously developed bioreactor systems for dioxin-contaminated water and soil. We have shown that a fungus, Pseudallescheria boydii (P. boydii), isolated from activated sludge treating wastewater that contained dioxins, has the ability to degrade highly chlorinated dioxins. A reaction product of octachlorinated dibenzo-p-dioxin (OCDD) was identified as heptachlorinated dibenzo-p-dioxin. Therefore, one of the pathways for degradation of OCDD by this fungus was predicted to be as follows: OCDD is transformed by dechlorination and then one of the remaining aromatic rings is oxidized. To apply P. boydii to on-site technologies (e.g., bioreactor systems), as well as in situ technologies, enzyme treatment using a dioxin-degrading enzyme from P. boydii needs to be developed because P. boydii is a weak pathogenic fungus, known to cause opportunistic infection. As a result, we have studied enzyme purification of nonchlorinated dioxin, namely, dibenzo-pdioxin (DD). However, we did not try to identify enzymes capable of degrading highly chlorinated dioxins. This study has elucidated a method of enzyme assay for measuring OCDD-degrading activity, and has attempted to purify OCDD-degrading enzymes from P. boydii using enzyme assay. In addition, as first step toward purifying 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), 2,3,7,8-TCDD degradation tests were carried out

  19. Purification and deodorization of structured lipids by short path distillation

    DEFF Research Database (Denmark)

    Xu, Xuebing; Jacobsen, Charlotte; Nielsen, Nina Skall


    Purification of structured lipids (SL), produced from lipase- catalyzed acidolysis of rapeseed oil and capric acid, and deodorization of randomized SL, produced from chemical randomization of fish oil and tricaprin, were studied in a bench-scale short path distillation (SPD). SL obtained from...

  20. Purification and characterization of protease from Bacillus cereus ...

    African Journals Online (AJOL)



    Sep 16, 2013 ... Purification and characterization of protease from. Bacillus cereus SU12 isolated from oyster. Saccostrea cucullata. S. Umayaparvathi*, S. Meenakshi, M. Arumugam and T. Balasubramanian. Centre of Advanced Study in Marine Biology, Faculty of Marine Sciences, Annamalai University, Parangipettai - 608.

  1. Kinetic modelling and thermodynamic studies on purification of ...

    African Journals Online (AJOL)


    toxicological and aesthetic reasons, polymer should in principle be as free as possible of monomer. This applies in particular to polymers for cosmetics and especially for pharmaceutical applications (Denzinger, et al., 1977). There are several previous research works aiming at PVP purification by adsorption (Nuber, et al., ...

  2. Membrane concepts for blood purification : towards improved artificial kidney devices

    NARCIS (Netherlands)

    Tijink, M.S.L.


    The research presented in this thesis is about the fabrication and characterization of new membranes for blood purification. A novel membrane concept is proposed to combine diffusion and adsorption in one step to remove uremic retention solutes. A membrane with embedded functionalized particles, a

  3. Methods in elastic tissue biology: elastin isolation and purification. (United States)

    Mecham, Robert P


    Elastin provides recoil to tissues subjected to repeated stretch, such as blood vessels and the lung. It is encoded by a single gene in mammals and is secreted as a 60-70 kDa monomer called tropoelastin. The functional form of the protein is that of a large, highly crosslinked polymer that organizes as sheets or fibers in the extracellular matrix. Purification of mature, crosslinked elastin is problematic because its insolubility precludes its isolation using standard wet-chemistry techniques. Instead, relatively harsh experimental approaches designed to remove non-elastin 'contaminates' are employed to generate an insoluble product that has the amino acid composition expected of elastin. Although soluble, tropoelastin also presents problems for isolation and purification. The protein's extreme stickiness and susceptibility to proteolysis requires careful attention during purification and in tropoelastin-based assays. This article describes the most common approaches for purification of insoluble elastin and tropoelastin. It also addresses key aspects of studying tropoelastin production in cultured cells, where elastin expression is highly dependent upon cell type, culture conditions, and passage number.

  4. New purification process of fungal immunomodulatory protein, FIP ...

    African Journals Online (AJOL)

    cy l

    parameters can be amplified linearly. Concentrated supernatant from ultrafiltration was further separated with two-step chromatographic purification of strong cation-exchange and size-exclusion, purified product shows as single band at 13 kDa in SDS-PAGE gel, the analysis result of reversed phase HPLC C4 column also ...

  5. Purification, characterization and ELISA detection of mink immunoglobulins

    DEFF Research Database (Denmark)

    Martel, Cyril Jean-Marie; Aasted, Bent


    This study describes easy purification methods for mink IgG, IgA and IgM immunoglobulins. IgG and IgM were purified from normal mink serum, while IgA was purified from mink bile from healthy animals. By SDS-polyacrylamid-gel-electrophoresis  (SDS-PAGE) and immunoblotting under reducing conditions...

  6. Potential for sustainable energy with biogas from sewage purification

    International Nuclear Information System (INIS)

    Coenen, J.; Van Gastel, M.; De Jong, K.


    Insight is given into the possibility to produce biogas from sewage purification plants in the Netherlands. Attention is paid to the estimated potential of sustainable energy from biogas, the economic effectiveness of several scenarios, the critical success factors and bottlenecks [nl

  7. Production and purification of polyclonal antibody against bovine ...

    African Journals Online (AJOL)

    Antibodies are important tools in medical researches which have led to many advances in this field. Anti-bovine immunoglobulins and its conjugate with horse radish peroxidase (HRP) is used to diagnose cows' disease by ELISA or western blotting tests. In this study, the production, purification and horse radish peroxidase ...

  8. Purification and characterization of laccase from Trametes hirsuta ...

    African Journals Online (AJOL)

    Purification and characterization of laccase from Trametes hirsuta Bm-2 and its contribution to dye and effluent decolorization. P Zapata-Castillo, MdeL Villalonga-Santana, J Tamayo-Cortés, G Rivera-Muñoz, S Solís-Pereira ...

  9. Comparative study of peroxidase purification from apple and orange ...

    African Journals Online (AJOL)



    Jul 6, 2011 ... It was observed that after partial purification, the enzyme activity was ... present in higher plants like turnip, sweet potato, tomato, ... enzyme extracted. Thermal treatment. To selectively inactivate the contaminating traces of the catalase moieties, crude enzyme extract was heated at 65°C for 3 min in a.

  10. Testing of the influenza virus purification by CIEF

    Czech Academy of Sciences Publication Activity Database

    Horká, Marie; Kubíček, O.; Kubesová, Anna; Kubíčková, Z.; Rosenbergová, K.; Šlais, Karel


    Roč. 31, č. 2 (2010), s. 331-338 ISSN 0173-0835 R&D Projects: GA AV ČR IAAX00310701 Institutional research plan: CEZ:AV0Z40310501 Keywords : capillary isoelectric focusing * PCR * purification efficiency Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 3.569, year: 2010

  11. Purification of cadmium by selective volatilization in vacuum in ...

    Indian Academy of Sciences (India)

    As the degree of refinement increases to higher levels, purification methods are predominately based on physical processes like vacuum distillation, crystallization, diffusion, electro-migration, etc, (Singh et al 1968). The distillation at batch scale requires considerable control of experimen- tal parameters, which can play an ...

  12. Isolation and Purification of Sesquiterpene Lactones from Ixeris ...

    African Journals Online (AJOL)

    phase deactivation, and contamination, and has made great progress in preparative isolation and purification ... During HSCCC separation, a two-phase solvent system of ethyl acetate n-butanol methanol water (4 : 6 : 1 ... a flow rate of 2.0 mL/min, while the apparatus was rotated at 900 rpm. After hydrodynamic equilibrium ...

  13. Purification of cadmium up to 5N+ by vacuum distillation

    Indian Academy of Sciences (India)


    defence, military and aero-space (Lejbov et al 1989). Other areas of applications include telecommunications, electric vehicle and remote area storage systems. The problem connected with profound purification and characterization of the materials that are used for the synthesis of electronic materials remains critical. The.

  14. Purification and characterization of β-glucosidase from ...

    African Journals Online (AJOL)



    Dec 17, 2008 ... thermostable β-glucosidase activity from thermophilic fungus. Humicola grisea var. thermoidea: purifcation and biochemical characterization. FEMS Microbiol. Lett. 146: 291-295. Pontoh J, Low NH (2002). Purification and characterization of β- glucosidase from honey bees (Apis mellifera). Insect Biochem.

  15. Expression and Purification of Coat Protein of Citrus Tristeza Virus ...

    African Journals Online (AJOL)

    transformed to BL21™ star (DE3) of E. coli expression competent cell were also compared using discontinues SDS-PAGE. Large scale recombinant protein production and purification. Large scale recombinant protein production was conducted using four one liter flask containing 250 ml 2xYT broth media consists of 100 ...

  16. Purification of heat labile toxin from Bordetella pertussis vaccine ...

    African Journals Online (AJOL)

    K.C. Shivanandappa


    Jun 23, 2015 ... ... °C until further use. The condition of the run must be +4 °C because the pertussis pro- teins are heat labile. Three consecutive runs were conducted for each G50 experiment. 2.10. Purification of HLT. G50 purified fractions were analyzed for its qualities through optical absorbance, and total protein SDS.

  17. Expression and purification of recombinant Shiga toxin 2B from ...

    African Journals Online (AJOL)

    sunny t

    (SDS-PAGE) and StxB2 yield was 450 µg ml-1 confirmed by Bradford assay. Recombinant Stx2B protein was produced in highly pure yield using HaloTag technology. Key words: Escherichia coli O157:H7, StxB gene, expression, HaloTag technology, purification. INTRODUCTION. Enterohemorrhagic Escherichia coli ...

  18. Comparative study of methods for extraction and purification of ...

    African Journals Online (AJOL)



    Aug 2, 2010 ... (1997), wastewater sludge sample was prepared by low speed centrifugation and homogenization in buffer (EDTA) and cell walls were lysed by bead-beating and chemical lysis (SDS, 65°C, 1h). Proteins were removed by phenol : chloroform: isoamylalcohol (25:24:1). DNA purification was conducted.

  19. Extraction and Purification of Flavonoids from Radix Puerariae | Li ...

    African Journals Online (AJOL)

    Purpose: To develop an efficient method for the purification of flavonoids from Radix puerariae. Methods: Optimal extraction technology was obtained using orthogonal test. Through adsorption and desorption tests, 8 resins with different polarity, diameter, and surface area were studied. Finally, a novel macroporous resin, ...

  20. Fungal laccase: copper induction, semi-purification, immobilization ...

    African Journals Online (AJOL)

    Fungal laccase: copper induction, semi-purification, immobilization, phenolic effluent treatment and electrochemical measurement. ... In order to apply in an effluent treatment, laccase was immobilized on different vitroceramics supports, pyrolytic graphite and also on a carbon fiber electrode as biosensor. The maximum ...

  1. Purification of recombinant C-terminus polyhistidine tagged human ...

    African Journals Online (AJOL)



    . ... E. coli TOP 10F' and P. pastoris KM71H (arg4 aox1::ARG4) strains. E. coli TOP 10F' and P. pastoris KM71H ..... I-guided inquiry-purification and characterization of a fusion protein: Histidine tag, malate dehydrogenase, and ...

  2. Purification and characterization of a soluble calnexin from human placenta

    DEFF Research Database (Denmark)

    Olsen, Dorthe T; Peng, Li; Træholt, Sofie D


    Calreticulin (Crt) and calnexin (Cnx) are homologous endoplasmic reticulum (ER) chaperones involved in protein folding and quality control. Crt is a soluble ER luminal Mr 46 kDa protein and Cnx is a Mr 67kDa ER membrane protein. During purification of Crt from human placenta a soluble form of Cnx...

  3. Purification of HGI2 for nuclear detector fabrication

    International Nuclear Information System (INIS)

    Schieber, M.M.


    A process for purification of mercuric iodide (HgI 2 ) to be used as a source material for the growth of detector quality crystals. The high purity HgI 2 raw material is produced by a combination of three stages: synthesis of HgI 2 from Hg and I 2 , repeated sublimation, and zone refining

  4. 21 CFR 876.5665 - Water purification system for hemodialysis. (United States)


    ... dialysate. This generic type of device may include a water softener, sediment filter, carbon filter, and... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Water purification system for hemodialysis. 876... SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5665 Water...

  5. The technology of large-scale pharmaceutical plasmid purification ...

    African Journals Online (AJOL)

    Further test demonstrated that the pcDNAlacZ purified with CTAB and authoritative endotoxin-free plasmid Kit had the similar transfection efficiency in vivo and in vitro. CTAB can be used for plasmid purification; the main advantages of the DNAs purified with CTAB include the avoidance of animal-derived enzymes, toxic ...

  6. Purification of kappa (k)-carrageenase from locally isolated ...

    African Journals Online (AJOL)

    Partial purification of the crude kappa (k)-carrageenase present in the culture filtrates of Cellulosimicrobium cellulans was carried out by fractional precipitation, using ammonium sulphate, acetone and ethanol individually. The highest recovered protein (37.08%) combined with enzyme activity was obtained with ammonium ...

  7. A study on the extraction and purification process of lily ...

    African Journals Online (AJOL)

    The objective of this paper was to extract and purify lily polysaccharide and to study its anti-H22 hepatoma effect in mice. Orthogonal experimental method was used to analyze the factors influencing the extraction and purification of lily polysaccharide, and the anti-tumor effect of lily polysaccharide was studied by acting it on ...

  8. Purification and characterization of angiotensin-1 converting enzyme

    African Journals Online (AJOL)

    Purification yield of the active peptide was estimated to be 0.2 ± 0.1%, starting from the lyophilized jellyfish. The infrared (IR), proton nuclear magnetic resonance spectroscopy (1H NMR), carbon nuclear magnetic resonance (13C NMR) and mass spectrometry (MS) spectrometer analyses elucidated that the structure of the ...

  9. Melting and purification methods in electron beam furnaces. Pt. 1

    International Nuclear Information System (INIS)

    Stephan, H.


    After a general review on the market of electron beam furnaces, the melting methods and their variations are discussed in detail. The purification effects shown in a table allow a view on the main application. The losses of volatile alloy elements and the chances of compensation are specially discussed. (orig.) [de

  10. High-level expression, purification, polyclonal antibody preparation ...

    African Journals Online (AJOL)



    Feb 14, 2011 ... OprD is a specific porin which can binds imipenem and carbapenems in Pseudomonas aeruginosa. OprD loss plays a central role in mediating carbapenem resistance. Therefore, purification of oprD protein lays a pavement for the study in vivo and in vitro. In our study, the oprD gene was cloned into.

  11. Effective Purification of Biogas by Condensing-Liquid Membrane

    Czech Academy of Sciences Publication Activity Database

    Poloncarzová, Magda; Vejražka, Jiří; Veselý, Václav; Izák, Pavel


    Roč. 50, č. 3 (2010), s. 669-671 ISSN 1433-7851 R&D Projects: GA MPO FR-TI1/245 Institutional research plan: CEZ:AV0Z40720504 Keywords : biogas purification * condensing liquid * gas permeation Subject RIV: CI - Industrial Chemistry, Chemical Engineering Impact factor: 12.730, year: 2010

  12. Production and purification of polyclonal antibody against bovine ...

    African Journals Online (AJOL)



    Jun 18, 2007 ... that ion-exchange chromatography could be an appropriate method for purification of IgG antibodies. Key words: Anti bovine immunoglobulins, horse radish peroxidase conjugation, ion-exchange chromatography, polyclonal ... diagnostic and therapeutic applications (Gallacher, 1993;. Gathumbi et al.

  13. High-level expression, purification, polyclonal antibody preparation ...

    African Journals Online (AJOL)

    OprD is a specific porin which can binds imipenem and carbapenems in Pseudomonas aeruginosa. OprD loss plays a central role in mediating carbapenem resistance. Therefore, purification of oprD protein lays a pavement for the study in vivo and in vitro. In our study, the oprD gene was cloned into pQE30 expression ...

  14. Partial purification and characterization of alkaline proteases from ...

    African Journals Online (AJOL)

    Alkaline proteases from the digestive tract of anchovy were partially purified by ammonium sulfate fractionation, dialysis and Sephadex G-75 gel filtration. The purification fold and yield were 6.23 and 4.49%, respectively. The optimum activities of partially purified alkaline proteases were observed at 60°C and at pH 11.0.

  15. Production and purification of polyclonal anti-hamster ...

    African Journals Online (AJOL)



    Jan 17, 2011 ... Purified antibody and its conjugation with HRP are used in research and diagnosis of hamster disease. Key words: Production, purification, hamster immunoglobulins. INTRODUCTION. Polyclonal antibodies represent a group or mixture of antibodies produced by different B-lymphocytes in res- ponse to the ...

  16. Cloning, high-level expression, purification and characterization of a ...

    African Journals Online (AJOL)

    The staphylokinase (Sak) is emerging as an important thrombolytic agent for the treatment of patients suffering from cardiovascular disease. Hence in this study, we reported the cloning, high-level expression, purification and characterization of the Sak variant SakøC from Staphylococcus aureus QT08 in Escherichia coli ...

  17. Purification and characterization of camel liver L-asparaginase ...

    African Journals Online (AJOL)

    Purification and characterization of camel liver L-asparaginase. TM Maharem, GM Sabry, MR Mohamed, MA Emam. Abstract. L-asparaginase from camel liver was isolated and purified by heat denaturation followed by QAE-Sephadex A-50 column chromatography and SP-Sepharose column chromatography. The purified ...

  18. Purification performances of common reed beds based on the ...

    African Journals Online (AJOL)



    Nov 30, 2013 ... Objective: This study aims to determine the purification performances of common reed beds in order to facilitate ... water crops. According to the INSAE (2002), Benin had a population of 6 million people in 2000; in 2012, this population would reach 9 million. ..... Physics and Chemistry of the Earth, vol.

  19. Purification the surface of detail from biological contaminations

    International Nuclear Information System (INIS)

    Gabdrakhmanov, Az T; Israphilov, I H; Galiakbarov, A T; Gabdrakhmanov, Al T


    More than 70% of biodegradation occur due to the corrosion processes. A biological corrosion causes the greatest damage to the oil and gas-production industry, the Navy and pipelines, constructions of water supply, means of communication. This paper proposes an effective method of purification various surfaces from biological contaminations by using of cold plasma. (paper)

  20. Purification of adenoviral vectors using expanded bed chromatography. (United States)

    Peixoto, C; Ferreira, T B; Carrondo, M J T; Cruz, P E; Alves, P M


    The increasing numbers of pre-clinical and clinical trials where recombinant adenoviral vectors are used for gene therapy and vaccination require the development of cost-effective and reproducible large scale purification strategies of the biologically active particles. Alternatives to the traditional laboratory scale CsCl density gradient ultracentrifugation method, such as fixed bed chromatography strategies, have been developed, but the yields of final recovery remain too low due mainly to the capture and concentration steps taking place before and between the chromatographic stages. In this study, a rapid and efficient scale-able purification protocol allowing to obtain concentrated, pure and bioactive adenoviral vectors was developed. This allows efficient levels of binding to the column media and vector purification without centrifugation or filtration steps. Expanded bed chromatography followed by hollow fiber concentration allows the capture of viral particles directly from cellular extracts with high efficiency and vector purification is achieved in less than one working day with a minimal amount of sample handling, thus presenting an improvement over existing processes. The overall process yield reached 32%, representing an eight-fold improvement over results reported previously, while the purity is comparable to that obtained with the CsCl method.

  1. Purification and properties of cowpea mosaic virus RNA replicase

    NARCIS (Netherlands)

    Zabel, P.


    This thesis concerns the partial purification and properties of an RNA-dependent RNA polymerase (RNA replicase) produced upon infection of Vigna unguiculata plants with Cowpea Mosaic Virus (CPMV). The enzyme is believed to be coded, at least in part, by the virus genome and to

  2. Extraction and purification of formonometin from Trifolium pratense L ...

    African Journals Online (AJOL)

    Extraction and purification of formonometin from Trifolium pratense L: Physicochemical properties of its complex with lecithin. ... that there was no significant difference (p < 0.05) between the peak intensity of the physical mixture and the complex, while FT-IR analysis indicated interaction between formononetin and lecithin.

  3. Purification of bacteriocins using size-exclusion chromatography

    Directory of Open Access Journals (Sweden)

    Vivek K. Bajpai


    Full Text Available The bacteriocin purification involves following main steps. a. Extraction of cell-free-supernatant of bacteria. b. Ammonium sulfate precipitation. c. Dialysis. d. Diafiltration using PVP and e. Size-exclusion chromatography. However, depending on the nature of work, the compound could be further analyzed by reverse-phase HPLC, NMR, mass spectrometry and sequencing.

  4. Purification and Some Properties of a Thermostable α-Amylase ...

    African Journals Online (AJOL)

    This work reports the isolation, purification and some properties of a thermostable α-amylase producing Bacillus subtilis isolated from the soil. Soil samples were collected and screened for thermophilic bacterial strains with amylase activity and to examine the amylase heat tolerance potentiality. The isolate was Gram ...

  5. Rapid purification of high activity Taq DNA polymerase expressed in ...

    African Journals Online (AJOL)

    A simplified method is described here for the preparation of a thermostable Taq DNA polymerase enzyme from Escherichia coli (E. coli) strain DH5a carrying the pTTQ18 expression vector transformed with the Taq polymerase gene. Standard purifications were done with 1 litre batch cultures of E. coli cells and produced ...

  6. Affinity purification of polysaccharide degrading enzymes with crosslinked substrates

    NARCIS (Netherlands)

    Rozie, H.J.


    The aim of this work was to find economically favourable, affinity based, purification methods for several polysaccharide splitting bulk enzymes. The framework in which this study is done is described in Chapter 1.

    Chapter 2 describes the adsorption of endo-polygalacturonase

  7. Purification and characterization of the chitosanase from Aeromonas ...

    African Journals Online (AJOL)

    Purification and characterization of the chitosanase from Aeromonas sp. HG08. Y Sun, J Zhang, S Wang. Abstract. In this work, the characterization of a chitosanase-producing bacterium isolated from soil was reported and this strain was grouped under the genus Aeromonas by virtue of its morphological, physiological ...

  8. Purification of cadmium up to 5N+ by vacuum distillation

    Indian Academy of Sciences (India)

    Cadmium was refined by vacuum distillation, a technique suitable for low boiling and melting point materials, to remove the heavy and low vapour pressure impurities at ppm level. The detailed analysis of the purified Cd as well as raw Cd was done by ICP–OES techniques for 27 impurity elements. Purification was carried ...

  9. Purification of cadmium by selective volatilization in vacuum in ...

    Indian Academy of Sciences (India)

    Purification of cadmium in the presence of a thin (∼0.001 mm) oxide layer on the surface of the molten metal was carried out using a simple system designed and fabricated locally. The analytical results revealed that the distillation through oxide layer gave better separation for Co, Ni, Cu, Zn, Ag, Sn, Hg and Tl, when ...

  10. Fast and efficient protein purification using membrane adsorber systems. (United States)

    Suck, Kirstin; Walter, Johanna; Menzel, Frauke; Tappe, Alexander; Kasper, Cornelia; Naumann, Claudia; Zeidler, Robert; Scheper, Thomas


    The purification of proteins from complex cell culture samples is an essential step in proteomic research. Traditional chromatographic methods often require several steps resulting in time consuming and costly procedures. In contrast, protein purification via membrane adsorbers offers the advantage of fast and gentle but still effective isolation. In this work, we present a new method for purification of proteins from crude cell extracts via membrane adsorber based devices. This isolation procedure utilises the membranes favourable pore structure allowing high flow rates without causing high back pressure. Therefore, shear stress to fragile structures is avoided. In addition, mass transfer takes place through convection rather than diffusion, thus allowing very rapid separation processes. Based on this membrane adsorber technology the separation of two model proteins, human serum albumin (HSA) and immungluboline G (IgG) is shown. The isolation of human growth hormone (hGH) from chinese hamster ovary (CHO) cell culture supernatant was performed using a cation exchange membrane. The isolation of the enzyme penicillin acylase from the crude Escherichia coli supernatant was achieved using an anion exchange spin column within one step at a considerable purity. In summary, the membrane adsorber devices have proven to be suitable tools for the purification of proteins from different complex cell culture samples.

  11. Purification and properties of Rhizobial DehL expressed in ...

    African Journals Online (AJOL)



    Jun 17, 2008 ... Full Length Research Paper. Purification and properties of Rhizobial DehL expressed in Escherichia coli. Fahrul Huyop1*, Nooraini Abdul Rashid1, Roswanira A. B. Wahab2 and Ronald A. Cooper3. 1Industrial Biotechnology Department, University Technology Malaysia, 81310 Skudai, Johor, Malaysia.

  12. Synthesis Gas Purification Purification des gaz de synthèse

    Directory of Open Access Journals (Sweden)

    Chiche D.


    Full Text Available Fischer-Tropsch (FT based B-XTL processes are attractive alternatives for future energy production. These processes aim at converting lignocellulosic biomass possibly in co-processing with petcoke, coal, or vacuum residues into synthetic biofuels. A gasification step converts the feed into a synthesis gas (CO and H2 mixture , which undergoes the Fischer-Tropsch reaction after H2/CO ratio adjustment and CO2 removal. However synthesis gas also contains various impurities that must be removed in order to prevent Fischer-Tropsch catalyst poisoning. Due to the large feedstocks variety that can be processed, significant variations of the composition of the synthesis gas are expected. Especially, this affects the nature of the impurities that are present (element, speciation, as well as their relative contents. Moreover, due to high FT catalyst sensitivity, severe syngas specifications regarding its purity are required. For these reasons, synthesis gas purification constitutes a major challenge for the development of B-XTL processes. In this article, we focus on these major hurdles that have to be overcome. The different kinds of syngas impurities are presented. The influence of the nature of feedstocks, gasification technology and operating conditions on the type and content of impurities is discussed. Highlight is given on the fate of sulfur compounds, nitrogen compounds, halides, transition and heavy metals. Main synthesis gas purification technologies (based on adsorption, absorption, catalytic reactions, etc. are finally described, as well as the related challenges. Les procédés de synthèse de biocarburants par voie Fischer-Tropsch (FT, voies B-XTL, représentent des alternatives prometteuses pour la production d’énergie. Ces procédés permettent la conversion en carburants de synthèse de biomasse lignocellulosique, éventuellement mise en oeuvre en mélange avec des charges fossiles telles que petcoke, charbons ou résidus sous vide. Pour


    Directory of Open Access Journals (Sweden)

    Yu. P. Sedlukho


    Full Text Available The paper considers problems and features of biochemical removal of hydrogen sulfide from ground water. The analysis of existing methods for purification of ground water from hydrogen sulfide has been given in the paper. The paper has established shortcomings of physical and chemical purification of ground water. While using aeration methods for removal of hydrogen sulfide formation of colloidal sulfur that gives muddiness and opalescence to water occurs due to partial chemical air oxidation. In addition to this violation of sulfide-carbonate equilibrium taking place in the process of aeration due to desorption of H2S and CO2, often leads to clogging of degasifier nozzles with formed CaCO3 that causes serious operational problems. Chemical methods require relatively large flow of complex reagent facilities, storage facilities and transportation costs.In terms of hydrogen sulfide ground water purification the greatest interest is given to the biochemical method. Factors deterring widespread application of the biochemical method is its insufficient previous investigation and necessity to execute special research in order to determine optimal process parameters while purifying groundwater of a particular water supply source. Biochemical methods for oxidation of sulfur compounds are based on natural biological processes that ensure natural sulfur cycle. S. Vinogradsky has established a two-stage mechanism for oxidation of hydrogen sulfide with sulfur bacteria (Beggiatoa. The first stage presupposes oxidation of hydrogen sulphide to elemental sulfur which is accumulating in the cytoplasm in the form of globules. During the second stage sulfur bacteria begin to oxidize intracellular sulfur to sulfuric acid due to shortage of hydrogen sulfide.The paper provides the results of technological tests of large-scale pilot plants for biochemical purification of groundwater from hydrogen sulfide in semi-industrial conditions. Dependences of water quality

  14. Replacement of fine particle purification filter of the PHT purification system - 15083

    International Nuclear Information System (INIS)

    Lee, D.S.


    The increase of chalk river unidentified deposit (CRUD), a particulate corrosion product in PHT (primary heat transport) system with increased operating years of a nuclear power plant causes: -) the problems of increased heavy water decomposition and deuterium formation reaction due to catalytic reaction with CRUD, -) damage to PHT pump seal due to a corrosion product, -) damage to fuel channel closure seal, and increased radiation exposure of workers due to elevated dose rate in steam generator water chamber. Wolsung unit 3 and 4 have replaced fine filter media in PHT purification system in phases reducing the pore size of the filter media (5 μm → 2 μm → 1 μm → 0.45 μm) to solve this problem. The phased replacement of fine filter media by the one with a smaller pore size reduced CRUD in PHT system significantly and also radiation dose rate in steam generator water chamber. Accordingly, many problems related to the aging of a plant (including increased radiation exposure of workers during outage, damage to mechanical seal of PHT pump) have been solved. (author)

  15. Uranium hexafluoride purification; Purificacao de hexafluoreto de uranio

    Energy Technology Data Exchange (ETDEWEB)

    Araujo, Eneas F. de


    Uranium hexafluoride might contain a large amount of impurities after manufacturing or handling. Three usual methods of purification of uranium hexafluoride were presented: selective sorption, sublimation, and distillation. Since uranium hexafluoride usually is contaminated with hydrogen fluoride, a theoretical study of the phase equilibrium properties was performed for the binary system UF{sub 6}-HF. A large deviation from the ideal solution behaviour was observed. A purification unity based on a constant reflux batch distillation process was developed. A procedure was established in order to design the re boiler, condenser and packed columns for the UF{sub 6}-HF mixture separation. A bench scale facility for fractional distillation of uranium hexafluoride was described. Basic operations for that facility and results extracted from several batches were discussed. (author)

  16. Chemical purification of lanthanides for low-background experiments (United States)

    Boiko, R. S.


    There are many potentially active isotopes among the lanthanide elements which are possible to use for low-background experiments to search for double β decay, dark matter, to investigate rare α and β decays. These kind of experiments require very low level of radioactive contamination, but commercially available compounds of lanthanides are always contamined by uranium, thorium, radium, potassium, etc. A simple chemical method based on liquid-liquid extraction has been applied for the purification of CeO2, Nd2O3 and Gd˙2O˙3 from radioactive traces. Detailed schemes of purification procedure are described. Measurements by using HPGe spectrometry demonstrate high efficiency in K, Ra, Th, U contaminations reduction on at least one order of magnitude.

  17. Technical project for a new water purification solution

    Directory of Open Access Journals (Sweden)

    Toma Adina


    Full Text Available This research is part of the RO-BG Cross-Border Cooperation Program, project “CLEANDANUBE”, MIS-ETC 653, which has finalised by providing a common strategy to prevent the Danube’s pollution technological risks with oil and oil products. This paper presents a new sustainable water purification solution. A short introduction will be offered and an overview regarding the research and new methods to greening the waste is provided. The theoretical aspects of the centrifugal separation phenomenon are studied and the preliminary project bases were established. The paper conveys the possible constructive variations and the technological implications of those. Ultimately, the technical project for a new water purification solution and conclusions with critical points encountered during the designing phase are presented.

  18. Purification process of recombinant monoclonal antibodies with mixed mode chromatography. (United States)

    Maria, Sophie; Joucla, Gilles; Garbay, Bertrand; Dieryck, Wilfrid; Lomenech, Anne-Marie; Santarelli, Xavier; Cabanne, Charlotte


    An innovative process to purify mAb from CHO cell culture supernatant was developed. This three-step process involved two mixed mode resins and an anion exchange membrane. We used a human IgG mixture to determine the optimal conditions for each purification step. Thereafter, the whole process was evaluated and improved for the purification of a recombinant mAb produced in the supernatant of CHO cells. Once optimized, yield and purity of 88% and 99.9%, respectively were comparable to those obtained in a conventional process based on a capture step using protein A. In addition, aggregates, HCPs and DNA levels in the purified fraction were below regulatory specifications. Then we used mass spectrometry to identify contaminating proteins in the antibody fraction in order to highlight the behavior of HCPs. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Study on a new water purification equipment with spiral lamellas (United States)

    Feng, X. R.


    A new water purification equipment was introduced, especially the section of spiral lamellas. Utilization of spiral lamellas made the sedimentation space reach to 100%, not only improving sedimentation efficiency and reducing the cover space, but also saving investment. Production test results showed that the new water purification equipment with spiral lamellas had characteristics of excellent treatment efficiency and high shock resistant capacity. As the treatment water volume was 240 m3/d, when the turbidity, CODMn and UV254 were 203 NTU, 1.90 mg/L and 0.030 cm-1 in raw water, they were 0.32 NTU, 0.72mg/L and 0.011 cm-1 respectively in effluent water, which could fully meet the drinking water hygiene requirement.

  20. Separation process design for isolation and purification of natural products

    DEFF Research Database (Denmark)

    Malwade, Chandrakant R.

    selection of separation techniques and operating conditions. The key factor in designing separation processes with multiple unit operations is to determine the synergy between them which in turn demands molecular level understanding of process streams. Therefore, the methodology is fortified with process......, thereby providing process information crucial for determining synergistic effects between different unit operations. In this work, the formulated methodology has been used to isolate and purify artemisinin, an antimalarial drug, from dried leaves of the plant Artemisia annua. A process flow sheet...... is generated consisting of maceration, flash column chromatography and crystallization unit operations for extraction, partial purification and final purification of artemisinin, respectively. PAT framework is used extensively to characterize the process streams at molecular level and the generated process...

  1. Antibody Fragments and Their Purification by Protein L Affinity Chromatography

    Directory of Open Access Journals (Sweden)

    Gustav Rodrigo


    Full Text Available Antibodies and related proteins comprise one of the largest and fastest-growing classes of protein pharmaceuticals. A majority of such molecules are monoclonal antibodies; however, many new entities are antibody fragments. Due to their structural, physiological, and pharmacological properties, antibody fragments offer new biopharmaceutical opportunities. In the case of recombinant full-length antibodies with suitable Fc regions, two or three column purification processes centered around Protein A affinity chromatography have proven to be fast, efficient, robust, cost-effective, and scalable. Most antibody fragments lack Fc and suitable affinity for Protein A. Adapting proven antibody purification processes to antibody fragments demands different affinity chromatography. Such technology must offer the unit operation advantages noted above, and be suitable for most of the many different types of antibody fragments. Protein L affinity chromatography appears to fulfill these criteria—suggesting its consideration as a key unit operation in antibody fragment processing.

  2. Drinking water purification in the Czech Republic and worldwide

    International Nuclear Information System (INIS)

    Krmela, Jan; Beckova, Vera; Vlcek, Jaroslav; Marhol, Milan


    The report is structured as follows: (i) Legislative (hygienic) requirements for technologies applied to drinking water purification with focus on uranium elimination; (ii) Technological drinking water treatment processes (settling, filtration, precipitation, acidification, iron and manganese removal) ; (iii) State Office for Nuclear Safety requirements for the operation of facilities to separate uranium from drinking water and for the handling of saturated ionexes from such facilities; (iv) Material requirements for the operation of ionex filters serving to separate uranium from drinking water; (v) Effect of enhanced uranium concentrations in drinking waters on human body; (vi) Uranium speciation in ground waters; (vii) Brief description of technologies which are used worldwide for uranium removal; (viii) Technologies which are usable and are used in the Czech Republic for drinking water purification from uranium; (ix) Inorganic and organic ion exchangers and sorbents. (P.A.)

  3. Experimental nested purification for a linear optical quantum repeater (United States)

    Chen, Luo-Kan; Yong, Hai-Lin; Xu, Ping; Yao, Xing-Can; Xiang, Tong; Li, Zheng-Da; Liu, Chang; Lu, He; Liu, Nai-Le; Li, Li; Yang, Tao; Peng, Cheng-Zhi; Zhao, Bo; Chen, Yu-Ao; Pan, Jian-Wei


    Quantum repeaters1-4 are essential elements for demonstrating global-scale quantum communication. Over the past few decades, tremendous efforts have been dedicated to implementing a practical quantum repeater5-10. However, nested purification1, the backbone of a quantum repeater, remains a challenge because the capacity for successive entanglement manipulation is still absent. Here, we propose and demonstrate an architecture of nested purification using spontaneous parametric downconversion sources11. A heralded entangled photon pair with higher fidelity is successfully purified from two copies of low-fidelity pairs that experience entanglement swapping and noisy channels. By delicately designing the optical circuits, double-pair emission noise is eliminated automatically and the purified state can be used for scalable entanglement connections to extend the communication distance. Combined with a quantum memory, our approach can be applied immediately in the implemention of a practical quantum repeater.

  4. Effect of biological and chemical preparations on peroxidase activity in leaves of tomato plants

    Directory of Open Access Journals (Sweden)

    Yulia Kolomiets


    Full Text Available In terms of treating tomato variety Chaika with chemical preparations with active substances if aluminum phosphate, 570 g/l + phosphorous acid 80 g/,l and mankotseb in concentration of 640 g/kg, the maximum increase in peroxidase activity in leaves of plants was observed in12 hours. In terms of use of biological preparations based on living cells Bacillus subtilis and Azotobacter chroococcum its activity was maximum in 24 hours and ranged from 77.7 to 112.7•s-1

  5. The Blood Compatibilities of Blood Purification Membranes and Other Materials Developed in Japan

    Directory of Open Access Journals (Sweden)

    Takaya Abe


    Full Text Available The biocompatibilities in blood purification therapy are defined as “a concept to stipulate safety of blood purification therapy by an index based on interaction in the body arising from blood purification therapy itself.” The biocompatibilities are associated with not only materials to be used but also many factors such as sterilization method and eluted substance. It is often evaluated based on impacts on cellular pathways and on humoral pathways. Since the biocompatibilities of blood purification therapy in particular hemodialysis are not just a prognostic factor for dialysis patients but a contributory factor for long-term complications, it should be considered with adequate attention. It is important that blood purification therapy should be performed by consistently evaluating not only risks associated with these biocompatibilities but also the other advantages obtained from treatments. In this paper, the biocompatibilities of membrane and adsorption material based on Japanese original which are used for blood purification therapy are described.

  6. Secretory immunoglobulin purification from whey by chromatographic techniques. (United States)

    Matlschweiger, Alexander; Engelmaier, Hannah; Himmler, Gottfried; Hahn, Rainer


    Secretory immunoglobulins (SIg) are a major fraction of the mucosal immune system and represent potential drug candidates. So far, platform technologies for their purification do not exist. SIg from animal whey was used as a model to develop a simple, efficient and potentially generic chromatographic purification process. Several chromatographic stationary phases were tested. A combination of two anion-exchange steps resulted in the highest purity. The key step was the use of a small-porous anion exchanger operated in flow-through mode. Diffusion of SIg into the resin particles was significantly hindered, while the main impurities, IgG and serum albumin, were bound. In this step, initial purity was increased from 66% to 89% with a step yield of 88%. In a second anion-exchange step using giga-porous material, SIg was captured and purified by step or linear gradient elution to obtain fractions with purities >95%. For the step gradient elution step yield of highly pure SIg was 54%. Elution of SIgA and SIgM with a linear gradient resulted in a step yield of 56% and 35%, respectively. Overall yields for both anion exchange steps were 43% for the combination of flow-through and step elution mode. Combination of flow-through and linear gradient elution mode resulted in a yield of 44% for SIgA and 39% for SIgM. The proposed process allows the purification of biologically active SIg from animal whey in preparative scale. For future applications, the process can easily be adopted for purification of recombinant secretory immunoglobulin species. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Purification and characterization of nitrilase from Fusarium solani IMI196840

    Czech Academy of Sciences Publication Activity Database

    Vejvoda, Vojtěch; Kubáč, David; Davidová, A.; Kaplan, Ondřej; Šulc, Miroslav; Šveda, Ondřej; Chaloupková, R.; Martínková, Ludmila


    Roč. 45, č. 7 (2010), s. 1115-1120 ISSN 1359-5113 R&D Projects: GA AV ČR IAA500200708; GA MŠk(CZ) LC06010; GA MŠk OC09046; GA ČR GD305/09/H008; GA MPO FT-TA5/043 Institutional research plan: CEZ:AV0Z50200510 Keywords : fusarium solani * nitrilase * purification Subject RIV: EE - Microbiology, Virology Impact factor: 2.648, year: 2010

  8. Two-Step Vapor/Liquid/Solid Purification (United States)

    Holland, L. R.


    Vertical distillation system combines in single operation advantages of multiple zone refining with those of distillation. Developed specifically to load Bridgman-Stockbarger (vertical-solidification) growth ampoules with ultrapure tellurium and cadmium, system, with suitable modifications, serves as material refiner. In first phase of purification process, ampoule heated to drive off absorbed volatiles. Second phase, evaporator heated to drive off volatiles in charge. Third phase, slowly descending heater causes distillation from evaporator to growing crystal in ampoule.

  9. Purification of radiolabeled RNA products using denaturing gel electrophoresis


    Adachi, Hironori; Yu, Yi-Tao


    This unit discusses a basic method for purification of radiolabeled RNAs using denaturing polyacrylamide gel electrophoresis. The method consists of a number of experimental procedures, including total RNA preparation from yeast cells, isolation of a specific RNA from total yeast RNA, RNA 3' terminal labeling using nucleotide (5’[32P]pCp) addition (via ligation), denaturing (8 M urea) polyacrylamide gel electrophoresis, and RNA extraction from the gel slice. Key points for achieving good elec...

  10. Effect of matrices on affinity purification of protein C. (United States)

    Kang, K; Ryu, D; Drohan, W N; Orthner, C L


    One of the critical problems in scale-up of affinity chromatography is the mechanical strength of the support matrix against pressure. Because the costs of both the gel matrix and the ligand for the affinity chromatography are very high, the reusability of gel matrices is directly related to the total production cost. In certain cases, where the source material is viscous (e.g., blood plasma), irreversible deformation of gel matrices can readily occur, necessitating severe constraints in the flow rate. Consequently, productivity is low.We have characterized the system parameters and investigated the performance of various matrices that are commercially available. The experimental system used for this study was the immunoaffinity purification of protein C (an anticoagulant protein) from human blood plasma. The support matrices studied were cross-linked agarose, polymethyl acrylic, cellulose, and polyvinyl alcohol polymers. The major system parameters studied were pressure tolerance, coupling efficiency, adsorption efficiency, and batch adsorption/desorption kinetics of protein C to/from the monoclonal antibody (MAb)-Matrix complex. In addition, the apparent equilibrium constant and bandwidth of the product concentration profile in the eluate were characterized by performing pulse tests.A methodology was developed for evaluating the immunoaffinity column performance for the separation of protein C. By utilizing the experimentally measured parameters, the flow rate limitation for each purification step was computed. Then, the purification performance of the matrices were evaluated in terms of productivity per unit time. Among the matrices tested, cellulose was superior in overall performance for the immunoaffinity purification of protein C using a 10 cm x 10 cm column.

  11. Isolation and purification of lysozyme from the hen egg white




    The aim of the research was the development of the method of lysozyme isolation from hen egg proteins. Lysozyme was isolated by differential heat denaturation of proteins with changing of the medium pH value, followed by neutralization, dialysis and additional purification by gel chromatography on Sephadex G-50. Activity was determined by bacteriolytic method (with Micrococcus lysodeikticus 4698 as a substrate). The enzyme purity and molecular mass were determined using SDS-electrophoresis an...

  12. Reverse osmosis membrane of high urea rejection properties. [water purification (United States)

    Johnson, C. C.; Wydeven, T. J. (Inventor)


    Polymeric membranes suitable for use in reverse osmosis water purification because of their high urea and salt rejection properties are prepared by generating a plasma of an unsaturated hydrocarbon monomer and nitrogen gas from an electrical source. A polymeric membrane is formed by depositing a polymer of the unsaturated monomer from the plasma onto a substrate, so that nitrogen from the nitrogen gas is incorporated within the polymer in a chemically combined form.

  13. Efficiency of different methods of extraction and purification of cytokinins

    Czech Academy of Sciences Publication Activity Database

    Hoyerová, Klára; Gaudinová, Alena; Malbeck, Jiří; Dobrev, Petre; Kocábek, Tomáš; Šolcová, Blanka; Trávníčková, Alena; Kamínek, Miroslav


    Roč. 67, č. 11 (2006), s. 1151-1159 ISSN 0031-9422 R&D Projects: GA AV ČR(CZ) IAA600380507; GA ČR GA206/02/0967 Institutional research plan: CEZ:AV0Z50510513; CEZ:AV0Z50380511 Keywords : Arabidopsis thaliana * Cytokinin extraction * Cytokinin purification Subject RIV: EF - Botanics Impact factor: 2.417, year: 2006

  14. Aqueous Chloride Operations Overview: Plutonium and Americium Purification/Recovery

    Energy Technology Data Exchange (ETDEWEB)

    Gardner, Kyle Shelton [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Kimball, David Bryan [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Skidmore, Bradley Evan [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)


    These are a set of slides intended for an information session as part of recruiting activities at Brigham Young University. It gives an overview of aqueous chloride operations, specifically on plutonium and americium purification/recovery. This presentation details the steps taken perform these processes, from plutonium size reduction, dissolution, solvent extraction, oxalate precipitation, to calcination. For americium recovery, it details the CLEAR (chloride extraction and actinide recovery) Line, oxalate precipitation and calcination.

  15. The Effects of Water Plants on the Creek Water Purification


    山本, 史子; 中野, 芳輔; 舟越, 保; 弓削, こずえ; Yamamoto, Fumiko; Nakano, Yoshisuke; Funakoshi, Tamotsu; Yuge, Kozue


    The activity of some creek water plants on water purification was studied. Five water plants collected for the experiment are locally called, hoteiaoi (Eichhornia crassipes), yoshi (Phragmites cornmunis), hishi (Trapa natans L.var. bispinosa), ukikusa (Spirodela polyrhi2a), suzumenohie (Paspalum Thunbergii). Additionally, anacalis originated outside of Japan was studied for comparison. The items of measured water quality were, water temperature, DO, EC, pH and NO3. Hoteiaoi was most effective...

  16. Influence of the drying method in chitosans purification step

    International Nuclear Information System (INIS)

    Fonseca, Ana C.M.; Batista, Jorge G.S.; Bettega, Antonio; Lima, Nelson B. de


    Currently, the study of extracellular biopolymers properties has received prominence for being easy extraction and purification. Chitosan has been an attractive proposition for applications in various fields such as engineering, biotechnology, medicine and pharmacology. For such applications, it is necessary purification of chitosan to obtain a product more concentrated and free of undesirable impurities. However, at this stage of the process of obtaining the biopolymer may occur morphological and physicochemical changes. This study evaluated the influence of the drying process after purification of a commercial chitosan sample and the importance of this step and its cost/benefit in applications requiring a high degree of purity. The method of drying influenced in the organoleptic properties and in the main characteristics of material. Analysis of the crystal structure by X-ray diffraction showed that the degree of crystallinity, X (%), in the purified chitosan samples was lower when compared with the unpurified sample. The degree of acetylation, DA (%), was analyzed by spectroscopy infrared with no significant changes on the three drying methods assessed, unlike the viscosimetric molecular weight, M v , determined by capillary viscometry. (author)

  17. [Extraction and purification technologies of total flavonoids from Aconitum tanguticum]. (United States)

    Li, Yan-Rong; Yan, Li-Xin; Feng, Wei-Hong; Li, Chun; Wang, Zhi-Min


    To optimize the extraction and purification technologies of total flavonoids from Aconitum tanguticum whole plant. With the content of total flavonoids as index, the optimum extraction conditions for the concentration, volume of alcohol, extracting time and times were selected by orthogonal optimized; Comparing the adsorption quantity (mg/g) and resolution (%), four kinds of macroporous adsorption resins including D101, AB-8, X-5 and XAD-16 were investigated for the enrichment ability of total flavonoids from Aconitum tanguticum; Concentration and pH value of sample, sampling amount, elution solvent and loading and elution velocity for the optimum adsorption resin were determined. The content of total flavonoids in Aconitum tanguticum was about 4.39%; The optimum extraction technique was 70% alcohol reflux extraction for three times,each time for one hour, the ratio of material and liquid was 1:10 (w/v); The optimum purification technology was: using XAD-16 macroporous resin, the initial concentration of total flavonoids of Aconitum tanguticum was 8 mg/mL, the sampling amount was 112 mg/g dry resin, the pH value was 5, the loading velocity was 3 mL/min, the elution solvent was 70% ethanol and the elution velocity was 5 mL/min. Under the optimum conditions, the average content of total flavonoids was raised from 4.39% to 46.19%. The optimum extraction and purification technologies for total flavonoids of Aconitum tanguticum were suitable for industrial production for its simplicity and responsibility.

  18. Nonlinear electrophoresis for purification of soil DNA for metagenomics. (United States)

    Engel, Katja; Pinnell, Lee; Cheng, Jiujun; Charles, Trevor C; Neufeld, Josh D


    Purification of microbial DNA from soil is challenging due to the co-extraction of humic acids and associated phenolic compounds that inhibit subsequent cloning, amplification or sequencing. Removal of these contaminants is critical for the success of metagenomic library construction and high-throughput sequencing of extracted DNA. Using three different composite soil samples, we compared a novel DNA purification technique using nonlinear electrophoresis on the synchronous coefficient of drag alteration (SCODA) instrument with alternate purification methods such as direct current (DC) agarose gel electrophoresis followed by gel filtration or anion exchange chromatography, Wizard DNA Clean-Up System, and the PowerSoil DNA Isolation kit. Both nonlinear and DC electrophoresis were effective at retrieving high-molecular weight DNA with high purity, suitable for construction of large-insert libraries. The PowerSoil DNA Isolation kit and the nonlinear electrophoresis had high recovery of high purity DNA suitable for sequencing purposes. All methods demonstrated high consistency in the bacterial community profiles generated from the DNA extracts. Nonlinear electrophoresis using the SCODA instrument was the ideal methodology for the preparation of soil DNA samples suitable for both high-throughput sequencing and large-insert cloning applications. Copyright © 2011 Elsevier B.V. All rights reserved.

  19. Composting Guilt: An Ecological Critique of Purification of Past Wrongdoing

    Directory of Open Access Journals (Sweden)

    Katharina von Kellenbach


    Full Text Available A review of the fast growing body of literature on transitional justice (Hayner 2011, political reconciliation (Philpott 2006, Verdejo 2009, forgiveness (Amstutz 2005, apology (Celermajer 2009, guilt (Barkan 2000 repentance (Schimmel 2002, evil (Meister 2011, moral repair (Walker 2006, and cultural memory (Borneman 2011, shows that the concept of purification has so far received little attention.  But the language of purification operates in the background of diverse practices such as the exhumation and ceremonial reburial of the dead (Desbois 2009, the call for truth commissions to document wrongdoing, and reparation campaigns that apologize and offer restitution (Diner 2007. At their best, truth and reconciliation commissions create cathartic moments (Greek: katharos = pure and facilitate “performative transformations” that cleanse relations between perpetrators and victims (Cole 2010, 15. My new project, still in its early stages, asks whether the concept and ritual practice of purification can be used to enhance moral repair in individuals and to serve the restoration of social order in the aftermath of atrocity and systemic human rights abuses.

  20. Different purification methods and quality of sunflower biodiesel

    Energy Technology Data Exchange (ETDEWEB)

    Pighinelli, A.L.M.T.; Park, K.J. [Campinas State Univ., Sao Paulo (Brazil). School of Agricultural Engineering; Ferrari, R.A.; Miguel, A.M.R.O. [Food Technology Inst., Sao Paulo (Brazil)


    Biodiesel is derived from triacylglycerides and is produced primarily through transesterification, a chemical reaction of vegetable oils with alcohol, methanol or ethanol. The cost of raw material should be considered since 85 per cent of production cost is related to vegetable oil. The purpose of this study was to evaluate oil expression of sunflower seed. It also examined the sunflower crude oil as a raw material for biodiesel by transesterification in both laboratory and pilot scale studies. Three different biodiesel purification methods were examined. The best result for oil expelling (68.4 per cent) at the experimental stage was obtained for seeds with a moisture content of 6.9 per cent at 25 degrees C and at a screw speed of 114 rpm. For biodiesel production at the laboratory scale, the best result for oil expelling was 87.5 per cent. It was obtained with an ethanol:oil molar ratio of 4.7:1 and with a 4.42 per cent catalyst concentration related to the quantity of oil that had to be transesterified. The experimental condition was applied at a bigger scale with a batch stirred tank reactor. For purification with washing, the biodiesel yield was 84.2 per cent. Purification with silica resulted in a yield of 84.6 per cent. A better quality biofuel was obtained through distillation of biodiesel.

  1. Countercurrent tangential chromatography for large-scale protein purification. (United States)

    Shinkazh, Oleg; Kanani, Dharmesh; Barth, Morgan; Long, Matthew; Hussain, Daniar; Zydney, Andrew L


    Recent advances in cell culture technology have created significant pressure on the downstream purification process, leading to a "downstream bottleneck" in the production of recombinant therapeutic proteins for the treatment of cancer, genetic disorders, and cardiovascular disease. Countercurrent tangential chromatography overcomes many of the limitations of conventional column chromatography by having the resin (in the form of a slurry) flow through a series of static mixers and hollow fiber membrane modules. The buffers used in the binding, washing, and elution steps flow countercurrent to the resin, enabling high-resolution separations while reducing the amount of buffer needed for protein purification. The results obtained in this study provide the first experimental demonstration of the feasibility of using countercurrent tangential chromatography for the separation of a model protein mixture containing bovine serum albumin and myoglobin using a commercially available anion exchange resin. Batch uptake/desorption experiments were used in combination with critical flux data for the hollow fiber filters to design the countercurrent tangential chromatography system. A two-stage batch separation yielded the purified target protein at >99% purity with 94% recovery. The results clearly demonstrate the potential of using countercurrent tangential chromatography for the large-scale purification of therapeutic proteins. Copyright © 2010 Wiley Periodicals, Inc.

  2. Purification of Sardine Fish Oil Through Degumming and Neutralization

    Directory of Open Access Journals (Sweden)

    Stephanie Bija


    Full Text Available The quality of sardine fish oil can be improved by purification method through the step of degumming and neutralization. The aimed of this this study was analysis characteristic of crude sardin fish oil and determined the best method of purification. Degumming was carried out using 30% water and salt at concentration 5%, 8%, 10% b/v. Neutralization process using  NaOH with 16°Be and bleaching using 5% Magnesol XL. All step of refining was done at 50°C, 60°C, 70°C, and 80°C. The result of analysis showed that sardine crude fish oil had 24.86% of palmitic acid as the highest fatty acid, heavy metal was not detected,dencity was 0.92 g/cm3 and viscocity was 51 cPs. The best treatment of purification method was degumming using 5% NaCl at 50°C with rendement 65.37±0.72%; free fatty acid (FFA 0.38±0.03%; peroxide (PV 1.07±0.12 mEq/kg; anisidine (p-AnV 15.18±0.16 mEq/kg; total oxidation value (TOTOX 17.31±0.39 mEq/kg; and clarity was 75.09± 1.20%.

  3. Antimicrobial biosurfactants from marine Bacillus circulans: extracellular synthesis and purification. (United States)

    Mukherjee, S; Das, P; Sivapathasekaran, C; Sen, R


    To purify the biosurfactant produced by a marine Bacillus circulans strain and evaluate the improvement in surface and antimicrobial activities. The study of biosurfactant production by B. circulans was carried out in glucose mineral salts (GMS) medium using high performance thin layer chromatography (HPTLC) for quantitative estimation. The biosurfactant production by this strain was found to be growth-associated showing maximum biosurfactant accumulation at 26 h of fermentation. The crude biosurfactants were purified using gel filtration chromatography with Sephadex G-50 matrix. The purification attained by employing this technique was evident from UV-visible spectroscopy and TLC analysis of crude and purified biosurfactants. The purified biosurfactants showed an increase in surface activity and a decrease in critical micelle concentration values. The antimicrobial action of the biosurfactants was also enhanced after purification. The marine B. circulans used in this study produced biosurfactants in a growth-associated manner. High degree of purification could be obtained by using gel filtration chromatography. The purified biosurfactants showed enhanced surface and antimicrobial activities. The antimicrobial biosurfactant produced by B. circulans could be effectively purified using gel filtration and can serve as new potential drugs in antimicrobial chemotherapy.

  4. Purification of a Multidrug Resistance Transporter for Crystallization Studies

    Directory of Open Access Journals (Sweden)

    Kamela O. Alegre


    Full Text Available Crystallization of integral membrane proteins is a challenging field and much effort has been invested in optimizing the overexpression and purification steps needed to obtain milligram amounts of pure, stable, monodisperse protein sample for crystallography studies. Our current work involves the structural and functional characterization of the Escherichia coli multidrug resistance transporter MdtM, a member of the major facilitator superfamily (MFS. Here we present a protocol for isolation of MdtM to increase yields of recombinant protein to the milligram quantities necessary for pursuit of structural studies using X-ray crystallography. Purification of MdtM was enhanced by introduction of an elongated His-tag, followed by identification and subsequent removal of chaperonin contamination. For crystallization trials of MdtM, detergent screening using size exclusion chromatography determined that decylmaltoside (DM was the shortest-chain detergent that maintained the protein in a stable, monodispersed state. Crystallization trials of MdtM performed using the hanging-drop diffusion method with commercially available crystallization screens yielded 3D protein crystals under several different conditions. We contend that the purification protocol described here may be employed for production of high-quality protein of other multidrug efflux members of the MFS, a ubiquitous, physiologically and clinically important class of membrane transporters.

  5. Large-scale functional purification of recombinant HIV-1 capsid.

    Directory of Open Access Journals (Sweden)

    Magdeleine Hung

    Full Text Available During human immunodeficiency virus type-1 (HIV-1 virion maturation, capsid proteins undergo a major rearrangement to form a conical core that protects the viral nucleoprotein complexes. Mutations in the capsid sequence that alter the stability of the capsid core are deleterious to viral infectivity and replication. Recently, capsid assembly has become an attractive target for the development of a new generation of anti-retroviral agents. Drug screening efforts and subsequent structural and mechanistic studies require gram quantities of active, homogeneous and pure protein. Conventional means of laboratory purification of Escherichia coli expressed recombinant capsid protein rely on column chromatography steps that are not amenable to large-scale production. Here we present a function-based purification of wild-type and quadruple mutant capsid proteins, which relies on the inherent propensity of capsid protein to polymerize and depolymerize. This method does not require the packing of sizable chromatography columns and can generate double-digit gram quantities of functionally and biochemically well-behaved proteins with greater than 98% purity. We have used the purified capsid protein to characterize two known assembly inhibitors in our in-house developed polymerization assay and to measure their binding affinities. Our capsid purification procedure provides a robust method for purifying large quantities of a key protein in the HIV-1 life cycle, facilitating identification of the next generation anti-HIV agents.

  6. Preparative Purification of Liriodendrin from Sargentodoxa cuneata by Macroporous Resin (United States)

    Li, Di-Hua; Wang, Yan; Lv, Yuan-Shan; Liu, Jun-Hong; Yang, Lei; Zhang, Shu-Kun; Zhuo, Yu-Zhen


    The preparative purification of liriodendrin from Sargentodoxa cuneata using macroporous resin combined with crystallization process was evaluated. The properties of adsorption/desorption of liriodendrin on eight macroporous resins were investigated systematically. X-5 resin was selected as the most suitable medium for liriodendrin purification. The adsorption of liriodendrin on X-5 resin fitted well with the pseudo-second-order kinetic model and Langmuir isotherm model. Dynamic adsorption/desorption tests were performed using a glass column packed with X-5 resin to optimize the separation process of liriodendrin. After one treatment with X-5 resin, the content of liriodendrin in the product was increased 48.73-fold, from 0.85% to 41.42%, with a recovery yield of 88.9%. 97.48% liriodendrin was obtained by further crystallization and determined by HPLC. The purified product possessed strong antioxidant activity. In conclusion, purification of liriodendrin might expend its further pharmacological researches and further applications in pharmacy. PMID:26236742

  7. Biomimetic small peptide functionalized affinity monoliths for monoclonal antibody purification. (United States)

    Wang, Xiangyu; Xia, Donghai; Han, Hai; Peng, Kun; Zhu, Peijie; Crommen, Jacques; Wang, Qiqin; Jiang, Zhengjin


    The rapid development of monoclonal antibodies (mAbs) in therapeutic and diagnostic applications has necessitated the advancement of mAbs purification technologies. In this study, a biomimetic small peptide ligand 3,5-di-tert-butyl-4-hydroxybenzoic acid-Arg-Arg-Gly (DAAG) functionalized monolith was fabricated through a metal ion chelation-based multi-step approach. The resulting monolith showed good chromatographic performance. Compared with the Ni 2+ based IMAC monolith, the DAAG functionalized monolith exhibited not only excellent specificity but also higher dynamic binding capacity (DBC). The 10% DBC and 50% DBC for hIgG reached as high values as 26.0 and 34.6 mg/mL, respectively, at a ligand density of 8.8 μmol/mL, due to the high porosity and accessibility of the monolithic matrix. Moreover, the stability of the DAAG functionalized monolith in successive breakthrough experiments indicates that it has a promising potential for long-term use in mAbs purification. Finally, the DAAG functionalized monolith was successfully applied to the purification of trastuzumab or human immunoglobulin G (hIgG) from biological samples. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. A comprehensive review on biodiesel purification and upgrading

    Directory of Open Access Journals (Sweden)

    Hamed Bateni


    Full Text Available Serious environmental concerns regarding the use of fossil-based fuels have raised awareness regarding the necessity of alternative clean fuels and energy carriers. Biodiesel is considered a clean, biodegradable, and non-toxic diesel substitute produced via the transesterification of triglycerides with an alcohol in the presence of a proper catalyst. After initial separation of the by-product (glycerol, the crude biodiesel needs to be purified to meet the standard specifications prior to marketing. The presence of impurities in the biodiesel not only significantly affects its engine performance but also complicates its handling and storage. Therefore, biodiesel purification is an essential step prior to marketing. Biodiesel purification methods can be classified based on the nature of the process into equilibrium-based, affinity-based, membrane-based, reaction-based, and solid-liquid separation processes. The main adverse properties of biodiesel – namely moisture absorption, corrosiveness, and high viscosity – primarily arise from the presence of oxygen. To address these issues, several upgrading techniques have been proposed, among which catalytic (hydrodeoxygenation using conventional hydrotreating catalysts, supported metallic materials, and most recently transition metals in various forms appear promising. Nevertheless, catalyst deactivation (via coking and/or inadequacy of product yields necessitate further research. This paper provides a comprehensive overview on the techniques and methods used for biodiesel purification and upgrading.

  9. Design and function of biomimetic multilayer water purification membranes. (United States)

    Ling, Shengjie; Qin, Zhao; Huang, Wenwen; Cao, Sufeng; Kaplan, David L; Buehler, Markus J


    Multilayer architectures in water purification membranes enable increased water throughput, high filter efficiency, and high molecular loading capacity. However, the preparation of membranes with well-organized multilayer structures, starting from the nanoscale to maximize filtration efficiency, remains a challenge. We report a complete strategy to fully realize a novel biomaterial-based multilayer nanoporous membrane via the integration of computational simulation and experimental fabrication. Our comparative computational simulations, based on coarse-grained models of protein nanofibrils and mineral plates, reveal that the multilayer structure can only form with weak interactions between nanofibrils and mineral plates. We demonstrate experimentally that silk nanofibril (SNF) and hydroxyapatite (HAP) can be used to fabricate highly ordered multilayer membranes with nanoporous features by combining protein self-assembly and in situ biomineralization. The production is optimized to be a simple and highly repeatable process that does not require sophisticated equipment and is suitable for scaled production of low-cost water purification membranes. These membranes not only show ultrafast water penetration but also exhibit broad utility and high efficiency of removal and even reuse (in some cases) of contaminants, including heavy metal ions, dyes, proteins, and other nanoparticles in water. Our biomimetic design and synthesis of these functional SNF/HAP materials have established a paradigm that could lead to the large-scale, low-cost production of multilayer materials with broad spectrum and efficiency for water purification, with applications in wastewater treatment, biomedicine, food industry, and the life sciences.

  10. Emulsion Liquid Membrane Technology in Organic Acid Purification

    International Nuclear Information System (INIS)

    Norela Jusoh; Norasikin Othman; Nur Alina Nasruddin


    Emulsion Liquid Membrane (ELM) process have shown a great potential in wide application of industrial separations such as in removal of many chemicals, organic compounds, metal ions, pollutants and biomolecules. This system promote many advantages including simple operation, high selectivity, low energy requirement, and single stage extraction and stripping process. One potential application of ELM is in the purification of succinic acid from fermentation broth. This study outline steps for developing emulsion liquid membrane process in purification of succinic acid. The steps include liquid membrane formulation, ELM stability and ELM extraction of succinic acid. Several carrier, diluent and stripping agent was screened to find appropriate membrane formulation. After that, ELM stability was investigated to enhance the recovery of succinic acid. Finally, the performance of ELM was evaluated in the extraction process. Results show that formulated liquid membrane using Amberlite LA2 as carrier, palm oil as diluent and sodium carbonate, Na 2 CO 3 as stripping agent provide good performance in purification. On the other hand, the prepared emulsion was observed to be stable up to 1 hour and sufficient for extraction process. In conclusion, ELM has high potential to purify succinic acid from fermentation broth. (author)

  11. Identification and Purification of Montmorillonite Mineral of Ghaen Mine Bentonite

    Directory of Open Access Journals (Sweden)

    P. Mirhoseini Moosavi


    Full Text Available Montmorillonite is the major mineral of Bentonite with many applications in industrial fields but some impurities decreases the quality of the bentonite. The main objective of this study was to investigate the suitable method for purification of Ghaen mine bentonite. A combination of methods was considered including wet sieving and sedimentation, centrifuge and ultrasound. The efficiency of purification methods was determined based on X-ray, particle size, cation exchange capacity (CEC and ratio peak of the Quartz/Montmorillonite analysis before and after experiments. The results showed that such methods were efficient for preparing of the materials having high quantity of montmorillonite with less than 2 microns particle sizes. Cristobalite was the only mineral remained in samples, however many of particles were exempted from the samples. Cristobalite was the main impurity remained with montmorillonite. Chemical treatment is the only way for its complete removal. The results of this study revealed that by using easy, cheap and fast methods, it is possible for acceptable purification of bentonite.



    Vovk, Oksana; Gay, Angela; Yakovleva, Anna


    Abstract. This article is devoted to the problem of municipal waste waters purification. The present daysituation with waste water treatment facilities in Ukraine, existed methods of waste waters purification andsearch for new ones are described. Much attention is paid to such kind of pollutants as microbiological andbacterial. A comparatively new method of sewage waters purification from biological contaminants andpossibilities to apply this method in Ukraine is presented in the article.Keyw...

  13. Operating experiences of gas purification system of Heavy Water Plant Talcher (Paper No. 1.11)

    International Nuclear Information System (INIS)

    Bhattacharya, R.; Mohanty, P.R.; Pandey, B.L.


    The operating experiences with the purification system installed at Heavy Water Plant, Talcher for purification of feed synthesis gas from fertilizer plant is described. The purification system has performed satisfactorily even with levels of impurities as much as 15 to 20 ppm of oxygen and carbon monoxide. The system could not however be tested at designed gas throughput and on a sustained basis. However, increase in gas throughput upto the design value is not expected to pose any problem on the performance of the purification system. (author). 5 figs

  14. Expression, purification, crystallization and preliminary X-ray analysis of Pseudomonas aeruginosa AlgX

    Energy Technology Data Exchange (ETDEWEB)

    Weadge, J.T.; Robinson, H.; Yip, P. P.; Arnett, K.; Tipton, P. A.; Howell, P. L.


    AlgX is a periplasmic protein required for the production of the exopolysaccharide alginate in Pseudomonas sp. and Azotobacter vinelandii. AlgX has been overexpressed and purified and diffraction-quality crystals have been grown using iterative seeding and the hanging-drop vapor-diffusion method. The crystals grew as flat plates with unit-cell parameters a = 46.4, b = 120.6, c = 86.9 {angstrom}, {beta} = 95.7{sup o}. The crystals exhibited the symmetry of space group P2{sub 1} and diffracted to a minimum d-spacing of 2.1 {angstrom}. On the basis of the Matthews coefficient (V{sub M} = 2.25 {angstrom}{sup 3} Da{sup -1}), two molecules were estimated to be present in the asymmetric unit.

  15. Purification of heat labile toxin from Bordetella pertussis vaccine strain 134 employed indigenous technology

    Directory of Open Access Journals (Sweden)

    K.C. Shivanandappa


    Conclusion: The B. pertussis HLT could be purified through two phase with G50 and DEAE, cost effective techniques, the G50 purification has reduced the bioburden problems during DEAE purification and at the same time the quality of the product was high.

  16. 24 CFR 203.52 - Acceptance of individual residential water purification equipment. (United States)


    ... have access to a continuing supply of safe and potable water without the use of a water purification... uninterrupted supply of safe and potable water adequate to meet household needs. (3) The water supply, when..., maintenance, repair and replacement of the water purification equipment. A copy of the signed service contract...

  17. Application of inorganic sorbents for waste waters purification in paper production

    International Nuclear Information System (INIS)

    Present work is devoted to application of inorganic sorbents for waste waters purification in paper production. Therefore as sorbents for waste waters purification of paper production inorganic materials (hydroxides of different elements, bentonite, chalk) are tested. The optimal conditions of sorption are defined.

  18. 21 CFR 884.6170 - Assisted reproduction water and water purification systems. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Assisted reproduction water and water purification... HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OBSTETRICAL AND GYNECOLOGICAL DEVICES Assisted Reproduction Devices § 884.6170 Assisted reproduction water and water purification systems. (a) Identification...

  19. Sodium tests on an integrated purification prototype for a sodium-cooled fast breeder reactor

    International Nuclear Information System (INIS)

    Abramson, R.


    This paper describes sodium tests performed on the integrated primary sodium purification prototype of the Creys Malville Super Phenix 1 fast breeder reactor. These tests comprised: - hydrostatic test, - thermal tests, - handling tests. They enabled a number of new technological arrangements to be qualified and provided the necessary information for the design and construction of the Super Phenix 1 purification units

  20. Item Purification Does Not Always Improve DIF Detection: A Counterexample with Angoff's Delta Plot (United States)

    Magis, David; Facon, Bruno


    Item purification is an iterative process that is often advocated as improving the identification of items affected by differential item functioning (DIF). With test-score-based DIF detection methods, item purification iteratively removes the items currently flagged as DIF from the test scores to get purified sets of items, unaffected by DIF. The…

  1. Technology of thorium concentrates purification and their transformation in pure nuclear products

    International Nuclear Information System (INIS)

    Ikuta, A.


    An experimental study for the purification of thorium concentrates by solvent extraction is presented. The product of purification is appropriate for utilization in the fabrication of nuclear reactor fuel elements. The experiments are carried out in a laboratory scale and the following operations are studied: dissolution, extraction-scrubbing, stripping-scrubbing, thorium oxalate precipitation, and thorium nitrate coagulation [pt

  2. Item Purification in Differential Item Functioning Using Generalized Linear Mixed Models (United States)

    Liu, Qian


    For this dissertation, four item purification procedures were implemented onto the generalized linear mixed model for differential item functioning (DIF) analysis, and the performance of these item purification procedures was investigated through a series of simulations. Among the four procedures, forward and generalized linear mixed model (GLMM)…

  3. Purification and Concentration of Nanoparticles Using Diafiltration: Scientific Operating Procedure Series: SOP-P-1 (United States)


    ER D C/ EL S R- 15 -4 Environmental Consequences of Nanotechnologies Purification and Concentration of Nanoparticles Using... Environmental Consequences of Nanotechnologies ERDC/EL SR-15-4 July 2015 Purification and Concentration of Nanoparticles Using Diafiltration...Methods of the ERDC/EL Environmental Consequences of Nanotechnologies research program. The primary goal of this Task was to develop robust SOPs for

  4. Purification of a Recombinant Glutathione Transferase from the Causative Agent of Hydatidosis, "Echinococcus granulosus" (United States)

    Fleitas, Andrea L.; Randall, Lía M.; Möller, Matías N.; Denicola, Ana


    This practical class activity was designed to introduce students to recombinant protein expression and purification. The principal goal is to shed light on basic aspects concerning recombinant protein production, in particular protein expression, chromatography methods for protein purification, and enzyme activity as a tool to evaluate purity and…

  5. Designing optimal bioethanol networks with purification for integrated biorefineries

    International Nuclear Information System (INIS)

    Shenoy, Akshay U.; Shenoy, Uday V.


    Highlights: • An analytical method is devised for bioethanol network integration with purification. • Minimum fresh bioethanol flow and pinch are found by the Unified Targeting Algorithm. • Optimal bioethanol networks are then synthesized by the Nearest Neighbors Algorithm. • Continuous targets and networks are developed over the purifier inlet flowrate range. • Case study of a biorefinery producing bioethanol from wheat shows large savings. - Abstract: Bioethanol networks with purification for processing pathways in integrated biorefineries are targeted and designed in this work by an analytical approach not requiring graphical constructions. The approach is based on six fundamental equations involving eight variables: two balance equations for the stream flowrate and the bioethanol load over the total network system; one equation for the above-pinch bioethanol load being picked up by the minimum fresh resource and the purified stream; and three equations for the purification unit. A solution strategy is devised by specifying the two variables associated with the purifier inlet stream. Importantly, continuous targeting is then possible over the entire purifier inlet flowrate range on deriving elegant formulae for the remaining six variables. The Unified Targeting Algorithm (UTA) is utilized to establish the minimum fresh bioethanol resource flowrate and identify the pinch purity. The fresh bioethanol resource flowrate target is shown to decrease linearly with purifier inlet flowrate provided the pinch is held by the same point. The Nearest Neighbors Algorithm (NNA) is used to methodically synthesize optimal networks matching bioethanol demands and sources. A case study of a biorefinery producing bioethanol from wheat with arabinoxylan (AX) coproduction is presented. It illustrates the versatility of the approach in generating superior practical designs with up to nearly 94% savings for integrated bioethanol networks, both with and without process

  6. Fractionation for Biodiesel Purification Using Supercritical Carbon Dioxide

    Directory of Open Access Journals (Sweden)

    Chao-Yi Wei


    Full Text Available In recent years, biodegradable and alternative biodiesel has attracted increased attention worldwide. Producing biodiesel from biomass involves critical separation and purification technology. Conventional technologies such as gravitational settling, decantation, filtration, water washing, acid washing, organic solvent washing and absorbent applications are inefficient, less cost effective and environmentally less friendly. In this study supercritical carbon dioxide (SC-CO2 with few steps and a low environmental impact, was used for biodiesel fractionation from impure fatty acid methyl ester (FAME solution mixes. The method is suitable for application in a variety of biodiesel production processes requiring subsequent stages of purification. The fractionation and purification was carried out using continuous SC-CO2 fractionation equipment, consisting of three columns filled with stainless steel fragments. A 41.85% FAME content solution mix was used as the raw material in this study. Variables were a temperature range of 40–70 °C, pressure range of 10–30 MPa, SC-CO2 flow rate range of 7–21 mL/min and a retention time range of 30–90 min. The Taguchi method was used to identify optimal operating conditions. The results show that a separated FAME content of 99.94% was verified by GC-FID under optimal fractionation conditions, which are a temperature of 40 °C of, a pressure level of 30MPa and a flow rate of 7 mL/min of SC-CO2 for a retention time of 90 min.

  7. Purification and characterization of alkaline proteases from aspergillus terreus

    International Nuclear Information System (INIS)

    Hussain, A.; Mannan, A.; Zubair, H.; Mirza, B.


    Proteases belong to an important class of enzymes known as hydrolases and catalyze hydrolysis of proteins. They act primarily to degrade proteins that are used for energy production and as biosynthetic precursors. In the following study, protease produced from Aspergillus terreus was found to be thermo stable and included in the category of alkaline serine and metallo protease. During partial purification, presence of enzyme in 60% (NH/sub 4/)/sub 2/SO/sub 4/ indicated small molecular weight polypeptide; later purification with Sephadex G-75 fractionation yielded a single proteolytic active molecule. At final purification step, the increase in specific activity of the enzyme was 7.5 fold with 23% yield. SDS-PAGE analysis revealed that alkaline protease of Aspergillus terreus is a monomer with approximate molecular weight of 35 kDa. Optimum pH for protease activity was found in the range of 7.5-11.0 (maximum at pH 8.5), thus apparently classified as an alkaline protease. The enzyme was thermo stable towards high temperature (60 deg. C), however it denatured irreversibly at 70 deg. C showing 80% loss of activity. The maximum proteolytic activity was found at 40 deg. C. The enzyme was effectively inhibited by PMSF, EDTA and urea whereas iodoacetamide and thiourea did not result in any loss in activity while cysteine was found to be activator molecule. The study with metal ions Mg/sup +2/, Mn/sup +2/ and Fe/sup +3/ (1 mM each) showed minute stimulatory effects on enzyme activity. Co/sup +2/ and Ca/sup +2/ (1 mM) had neither excitatory nor inhibitory effect while Hg/sup +2/ and Cu/sup +2/ (1 mM) slightly reduced the enzyme activity. (author)

  8. A Simple Slow-Sand Filter for Drinking Water Purification

    Directory of Open Access Journals (Sweden)

    K. O. Yusuf


    Full Text Available Water-borne diseases are commonly encountered when pathogen-contaminated water is consumed. In rural areas, water is usually obtained from ponds, open shallow wells, streams and rain water during rainy season. Rain water is often contaminated by pathogens due to unhygienic of physical and chemical conditions of the roofs thereby making it unsafe for consumption. A simple slow sand filter mechanism was designed and fabricated for purification of water in rural areas where electricity is not available to power water purification devices. Rain water samples were collected from aluminum roof, galvanized roof and thatched roof. The waters samples were allowed to flow through the slow sand filter. The values of turbidity, total dissolved solids, calcium, nitrite, faecal coliform and total coliform from unfiltered water through thatched roof were 0.92 NTU, 27.23 mg/l, 6 mg/l, 0.16 mg/l, 5cfu/100ml and 6.0 cfu/100ml, respectively while the corresponding values for slow sand filter from thatched roof were 0.01 NTU, 0.23 mg/l, 2.5 mg/l, 0.1 mg/l, 0 cfu/100ml and 0 cfu/100ml, respectively. The values of turbidity, total dissolved solid, nitrite, calcium, faecal coliform and total coliform from unfiltered water for aluminum roof were 0.82 NTU, 23.68 mg/l, 2.70 mg/l, 1.0 mg/l, 4 cfu/100ml and 4cfu/100ml, respectively while the corresponding values for slow sand filter were 0.01 NTU, 0.16 mg/l, 0.57 mg/l, 0.2 mg/l, 0 cfu/100ml and 0 cfu/100ml, respectively. The values obtained for galvanized roof were also satisfactory. The slow sand filter is recommended for used in rural areas for water purification to prevent risk of water-borne diseases.

  9. Anion exchange separation and purification of neodymium from fission products

    International Nuclear Information System (INIS)

    Ramkumar, K.L.; Raman, V.A.; Khodade, P.S.; Jain, H.C.


    Neodymium-148, the stable fission product has been proved to be one of the best monitors for the determination of nuclear fuel burn-up using triple spike isotope dilution mass spectrometry. For the precise and accurate determination of neodymium it is essential to separate it from bulk of other materials and purify from cerium and samarium which would otherwise cause isobaric interferences. A single stage anion exchange procedure for the separation and purification of neodymium from fission products has been developed. This method supercedes the lengthy and time consuming two stage anion exchange procedure normally used and ensures good chemical yield. (author)

  10. Recovering of thorium contained in wastes from Thorium Purification Plant

    International Nuclear Information System (INIS)

    Brandao Filho, D.; Hespanhol, E.C.B.; Baba, S.; Miranda, L.E.T.; Araujo, J.A. de.


    A study has been developed in order to establish a chemical process for recovering thorium from wastes produced at the Thorium Purification Plant of the Instituto de Pesquisas Energeticas e Nucleares. The recovery of thorium in this process will be made by means of solvent extraction technique. Solutions of TBP/Varsol were employed as extracting agent during the runs. The influence of thorium concentration in the solution, aqueous phase acidity, volume ratio of the phases, percentage of TBP/Varsol and the contact time of the phases on the extraction of thorium and lanthanides was determined. (author)

  11. Exploiting interfacial water properties for desalination and purification applications.

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Hongwu (Los Alamos National Laboratory, Los Alamos, NM); Varma, Sameer; Nyman, May Devan; Alam, Todd Michael; Thuermer, Konrad; Holland, Gregory P.; Leung, Kevin; Liu, Nanguo (University of New Mexico Albuquerque, NM); Xomeritakis, George K. (University of New Mexico Albuquerque, NM); Frankamp, Benjamin L.; Siepmann, J. Ilja (University of Minnesota, Minneapolis, MN); Cygan, Randall Timothy; Hartl, Monika A. (Los Alamos National Laboratory, Los Alamos, NM); Travesset, Alex (Iowa State University, Ames, IA); Anderson, Joshua A. (Iowa State University, Ames, IA); Huber, Dale L.; Kissel, David J. (University of New Mexico Albuquerque, NM); Bunker, Bruce Conrad; Lorenz, Christian Douglas; Major, Ryan C. (University of Minnesota, Minneapolis, MN); McGrath, Matthew J. (University of Minnesota, Minneapolis, MN); Farrow, Darcie; Cecchi, Joseph L. (University of New Mexico Albuquerque, NM); van Swol, Frank B.; Singh, Seema; Rempe, Susan B.; Brinker, C. Jeffrey; Clawson, Jacalyn S.; Feibelman, Peter Julian; Houston, Jack E.; Crozier, Paul Stewart; Criscenti, Louise Jacqueline; Chen, Zhu (University of New Mexico Albuquerque, NM); Zhu, Xiaoyang (University of Minnesota, Minneapolis, MN); Dunphy, Darren Robert (University of New Mexico Albuquerque, NM); Orendorff, Christopher J.; Pless, Jason D.; Daemen, Luke L. (Los Alamos National Laboratory, Los Alamos, NM); Gerung, Henry (University of New Mexico Albuquerque, NM); Ockwig, Nathan W.; Nenoff, Tina Maria; Jiang, Ying-Bing; Stevens, Mark Jackson


    A molecular-scale interpretation of interfacial processes is often downplayed in the analysis of traditional water treatment methods. However, such an approach is critical for the development of enhanced performance in traditional desalination and water treatments. Water confined between surfaces, within channels, or in pores is ubiquitous in technology and nature. Its physical and chemical properties in such environments are unpredictably different from bulk water. As a result, advances in water desalination and purification methods may be accomplished through an improved analysis of water behavior in these challenging environments using state-of-the-art microscopy, spectroscopy, experimental, and computational methods.

  12. Preparation and Characterization of Zeolite Membrane for Bioethanol Purification

    Directory of Open Access Journals (Sweden)

    Aprilina Purbasari


    Full Text Available The use of bioethanol as an alternative fuel with a purity of more than 99.5% wt has prompted research on bioethanol purification. One of the promising methods used for bioethanol purification is pervaporation membrane. This research is aimed to prepare and characterize zeolite membranes for pervaporation membrane. The membrane preparation consisted of two stages, namely support preparation and zeolite deposition on the support. In support preparation, α- alumina and kaolin with specific composition (50:30; 40:40; 50:30 was mixed with additives and water. After pugging and aging process, the mixture became paste and extruded into tubular shape. The tube was then calcined at temperature of 1250 °C for 3 hours. After that, zeolite 4A was deposited on the tubes using clear solution made of 10 %wt zeolite and 90 %wt water and heated at temperature of 80 °C for 3 hours. Furthermore, the resulting zeolite membranes was washed with deionized water for 5 minutes and dried in oven at temperature of 100 °C for 24 hours. Characterization of zeolite membranes included mechanical strength test, XRD, and SEM. In the mechanical strength test, the membrane sample with α- alumina:kaolin = 50:30 (membrane A has the highest mechanical strength of 46.65 N/mm2. Result of XRD analysis for the membrane A indicated that mullite and corundum phases were formed, which mullite phase was more dominant. Meanwhile the result of SEM analysis shows that zeolite crystals have been formed and covered the pores support, but the deposition of zeolite has not been optimal yet. The performance examination for bioethanol purification showed that the membrane could increase the purity of bioethanol from 95% to 98.5% wt. © 2013 BCREC UNDIP. All rights reservedReceived: 23rd October 2012; Revised: 15th February 2013; Accepted: 16th February 2013[How to Cite: Purbasari, A., Istirokhatun, T., Devi, A.M., Mahsunnah, L. , Susanto, H. (2013. Preparation and Characterization of Zeolite

  13. Systems and methods for separation and purification of products

    Energy Technology Data Exchange (ETDEWEB)

    Weiss, Michael Joseph; Gilliam, Ryan J.; Self, Kyle; Mariansky, Gal; Leclerc, Margarete K.; Shipchandler, Riyaz Mohammed; Nagar, Jacob


    There are provided methods and systems for an electrochemical cell including an anode and a cathode where the anode is contacted with a metal ion that converts the metal ion from a lower oxidation state to a higher oxidation state. The metal ion in the higher oxidation state is reacted with an unsaturated hydrocarbon and/or a saturated hydrocarbon to form products. Separation and/or purification of the products as well as of the metal ions in the lower oxidation state and the higher oxidation state, is provided herein.

  14. Fractionation and purification of the thiol proteinases from papaya latex. (United States)

    Dekeyser, P M; De Smedt, S; Demeester, J; Lauwers, A


    Three cysteine proteinases, i.e. chymopapain, papaya proteinase IV and proteinase III, were purified to homogeneity from papaya latex using a combination of ion-exchange chromatography and hydrophobic interaction chromatography. During the purification procedure, the thiol-groups of the active center were reversibly blocked as mixed disulfides with 2-thiopyridone. Homogeneity was proved electrophoretically by native polyacrylamide gel electrophoresis (PAGE), sodium dodecyl sulfate (SDS)-PAGE and rechromatography on a Mono S 5/5 column at pH 5.0.

  15. Novel Hydrogen Purification Device Integrated with PEM Fuel Cells

    Energy Technology Data Exchange (ETDEWEB)

    Joseph Schwartz; Hankwon Lim; Raymond Drnevich


    A prototype device containing twelve membrane tubes was designed, built, and demonstrated. The device produced almost 300 scfh of purified hydrogen at 200 psig feed pressure. The extent of purification met the program target of selectively removing enough impurities to enable industrial-grade hydrogen to meet purity specifications for PEM fuel cells. An extrusion process was developed to produce substrate tubes. Membranes met several test objectives, including completing 20 thermal cycles, exceeding 250 hours of operating life, and demonstrating a flux of 965 scfh/ft2 at 200 psid and 400 C.

  16. Chromatographic purification of tritiated steroids prior to use in radioimmunoassay

    International Nuclear Information System (INIS)

    Manlimos, F.S.; Abraham, G.E.


    The purity of tritiated steroids used as reagents in radioimmunoassay plays an important role in the reliability of the assay. These radioactive reagents should be assessed for purity upon receipt and the purity should be checked periodically afterward. For such purposes, we have used chromatographic purification on Celite microcolumns. By changing the polarity of the stationary and mobile phases, 20 different tritiated steroids with a wide range of polarity could be purified on these microcolumns. This approach is easy, rapid, economical, and reliable. (U.S.)

  17. Process for the biological purification of waste water

    DEFF Research Database (Denmark)


    Process for the biological purification of waste water by the activated sludge method, the waste water being mixed with recirculated sludge and being subjected to an anaerobic treatment, before the waste water thus treated is alternately subjected to anoxic and aerobic treatments and the waste...... water thus treated is led into a clarification zone for settling sludge, which sludge is recirculated in order to be mixed with the crude waste water. As a result, a simultaneous reduction of the content both of nitrogen and phosphorus of the waste water is achieved....

  18. Molecular Techniques for Dicistrovirus Detection without RNA Extraction or Purification

    Directory of Open Access Journals (Sweden)

    Jailson F. B. Querido


    Full Text Available Dicistroviridae is a new family of small, nonenveloped, and +ssRNA viruses pathogenic to both beneficial arthropods and insect pests as well. Triatoma virus (TrV, a dicistrovirus, is a pathogen of Triatoma infestans (Hemiptera: Reduviidae, one of the main vectors of Chagas disease. In this work, we report a single-step method to identify TrV, a dicistrovirus, isolated from fecal samples of triatomines. The identification method proved to be quite sensitive, even without the extraction and purification of RNA virus.

  19. Polyacrylamide Gel Electrophoresis for Purification of Large Amounts of RNA. (United States)

    Meyer, Mélanie; Masquida, Benoît


    Polyacrylamide gel electrophoresis (PAGE) constitutes a powerful technique for the efficient purification of RNA molecules dedicated to applications that require high purity levels. PAGE allows for the fractionation of RNA obtained from cell extracts, chemical or enzymatic synthesis, or modification experiments. Native or denaturing conditions can be chosen for analytical or preparative-scale separations and the nucleotide resolution can be tuned by changing the percentage and reticulation of the gel material. In this protocol, we focus on the preparation of milligram-scale amounts of ~200 nucleotides (nt) RNA molecules that were used in subsequent crystallization experiments.

  20. Purification of radiolabeled RNA products using denaturing gel electrophoresis. (United States)

    Adachi, Hironori; Yu, Yi-Tao


    This unit discusses a basic method for purification of radiolabeled RNAs using denaturing polyacrylamide gel electrophoresis. The method consists of a number of experimental procedures, including total RNA preparation from yeast cells, isolation of a specific RNA from total yeast RNA, RNA 3'-terminal labeling using nucleotide (5' [(32) P]pCp) addition (via ligation), denaturing (8 M urea) polyacrylamide gel electrophoresis, and RNA extraction from the gel slice. Key points for achieving good electrophoretic separation of RNA are also discussed. Copyright © 2014 John Wiley & Sons, Inc.

  1. Semiconductor Grade, Solar Silicon Purification Project. [photovoltaic solar energy conversion (United States)

    Ingle, W. M.; Rosler, R. S.; Thompson, S. W.; Chaney, R. E.


    A low cost by-product, SiF4, is reacted with mg silicon to form SiF2 gas which is polymerized. The (SiF2)x polymer is heated forming volatile SixFy homologues which disproportionate on a silicon particle bed forming silicon and SiF4. The silicon analysis procedure relied heavily on mass spectroscopic and emission spectroscopic analysis. These analyses demonstrated that major purification had occured and some samples were indistinguishable from semiconductor grade silicon (except possibly for phosphorus). However, electrical analysis via crystal growth reveal that the product contains compensated phosphorus and boron.

  2. Interim report on the TMI-2 purification filter examination

    International Nuclear Information System (INIS)

    Mason, R.E.; Hobbins, R.R.; Cook, B.A.; MacDonald, P.E.


    Filters from the purification/makeup system of the Three Mile Island Unit 2 Reactor were examined after the March 28, 1979, accident to determine the character of the debris transported to the filters. The general condition of the filters is presented. Material was removed from the filters and examined. The elemental and radionuclide makeup of the debris is discussed. Distribution of particle size and shape is presentd for some of the material examined. This is an interim report. When the investigation is completed, another report summarizing all of the data will be issued

  3. Expression, purification and characterization of human Dopamine ß-monooxygenase

    DEFF Research Database (Denmark)

    Vendelboe, Trine Vammen

    This thesis deals with expression, purification and characterization of the copper containing enzyme dopamine ß-monooxygenase (DBM). DBM is an ascorbate dependent protein that requires Cu in the active site in order to be functional. DBM is made of four domains; An Nterminal DOMON domain, the two...... others, one of the reasons why these proteins are considered to follow the same mechanism. DBM converts dopamine (DA) into Norepinphrine (NE). Both substrate and product functions as neurotransmitters and the levels of these are involved in many different disorders such as depression and hypertension...

  4. Purification and cloning of DNA fragments fractionated on agarose gels. (United States)

    Griffin, H G; Gasson, M J


    Purification of DNA fragments from acrylamide or agarose gels is a commonly used technique in the molecular biology laboratory. This article describes a rapid, efficient, and inexpensive method of purifying DNA fractions from an agarose gel. The purified DNA is suitable for use in a wide range of applications including ligation using DNA ligase. The procedure uses standard high-melting-temperature agarose and normal TBE electrophoresis buffer. In addition, the protocol does not involve the use of highly toxic organic solvents such as phenol.

  5. Preparation and purification of 7-Iodoclonazepam for use in radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Goddard, C.P.; Law, B.; Mason, P.A.; Stead, A.H.


    A method is described for the preparation and purification of 7-(/sup 125/I)-Iodoclonazepam (5-(o-Chlorophenyl)-2,3-Dihydro-7-(/sup 125/I)-Iodo-1H-1,4-Benzodiazepin-2-one). The structure was confirmed by mass spectrometry using 7-(/sup 127/I)iodoclonazepam prepared by the same method. 7-(/sup 125/I)-Iodoclonazepam binds well to a benzodiazepine antiserum. Although readily displaced by all the benzodiazepines commercially available in the UK, it is not displaced by structurally related nonbenzodiazepines except at very high concentrations. 7-(/sup 125/I) Iodoclonazepam should therefore be useful for the development of a screening radioimmunoassay (RIA) for benzodiazepines.

  6. Expression, purification and preliminary diffraction studies of PhnP

    DEFF Research Database (Denmark)

    Podzelinska, Kateryna; He, Shumei; Soares, Alexei


    PhnP belongs to a 14-gene operon that supports the growth of Escherichia coli on alkylphosphonates as a sole source of phosphorus; however, the exact biochemistry of phosphonate degradation by this pathway is poorly understood. The protein was recombinantly expressed in Escherichia coli and purif......PhnP belongs to a 14-gene operon that supports the growth of Escherichia coli on alkylphosphonates as a sole source of phosphorus; however, the exact biochemistry of phosphonate degradation by this pathway is poorly understood. The protein was recombinantly expressed in Escherichia coli...

  7. Magnetic purification of curcumin from Curcuma longa rhizome by novel naked maghemite nanoparticles. (United States)

    Magro, Massimiliano; Campos, Rene; Baratella, Davide; Ferreira, Maria Izabela; Bonaiuto, Emanuela; Corraducci, Vittorino; Uliana, Maíra Rodrigues; Lima, Giuseppina Pace Pereira; Santagata, Silvia; Sambo, Paolo; Vianello, Fabio


    Naked maghemite nanoparticles, namely, surface active maghemite nanoparticles (SAMNs), characterized by a diameter of about 10 nm, possessing peculiar colloidal stability, surface chemistry, and superparamagnetism, present fundamental requisites for the development of effective magnetic purification processes for biomolecules in complex matrices. Polyphenolic molecules presenting functionalities with different proclivities toward iron chelation were studied as probes for testing SAMN suitability for magnetic purification. Thus, the binding efficiency and reversibility on SAMNs of phenolic compounds of interest in the pharmaceutical and food industries, namely, catechin, tyrosine, hydroxytyrosine, ferulic acid, coumaric acid, rosmarinic acid, naringenin, curcumin, and cyanidin-3-glucoside, were evaluated. Curcumin emerged as an elective compound, suitable for magnetic purification by SAMNs from complex matrices. A combination of curcumin, demethoxycurcumin, and bis-demethoxycurcumin was recovered by a single magnetic purification step from extracts of Curcuma longa rhizomes, with a purity >98% and a purification yield of 45%, curcumin being >80% of the total purified curcuminoids.

  8. Small scale extraction and purification of human prolactin for the preparation of radioimmunoassay reagents

    International Nuclear Information System (INIS)

    Dias, L.E.M.F.


    Purification of human prolactin from pituitaries was carried out in our laboratory to obtain a pure reagent for use in RIA. The extraction and purification procedure was adapted from the method of Mc. Lean et al., and it involves the following steps: 1. Extraction of frozen pituitaries in buffers 0.14M phosphate/citrate pH 4.0 and 0.05M ammonium acetate pH 10.0. 2. Purification by hydrophobic interaction chromatography on Phenyl-Sepharose CL-4B in the presence of acetonitrile. 3. Purification by anion exchange chromatography on DEAE-Sepharose Cl-68. The purification method is considered effective for obtaining a hPrl of the purity needed for radioassay purposes, having the advantage of rapidity and relative simplicity. (author) [pt

  9. Determination of the self purification of streams using tracers

    International Nuclear Information System (INIS)

    Salviano, J.S.


    A methodology for the 'in situ' evaluation of the self purification of streams is discussed. It consists of the simultaneous injection of two tracers into the stream. One of the tracers is oxidized by biochemical processes. It can be either artificially supplied to the stream or a naturally present component can be used. This tracer is used for the determination of the self purification parameters. The other tracer is conservative and allows for the hydrodynamic effects. Tests have been carried out in two streams with quite different hydrodynamic and physicochemical conditions. In the first stream, with a flow-rate of about 0.9 m 3 /s, urea was used as the nonconservative tracer. In the other stream, which had a flow-rate of about 5 m 3 /s, only a radioactive tracer has been used, and the rate of biochemical oxidation has been determined from BOD measurements. Calculations have been implemented on a digital computer. In both cases it was found that the reoxygenation rate is more conveniently determined by empirical formulas. Results from both tests have been deemed realistic by comparison with similar experiments. (Author) [pt

  10. Cloning, purification and crystallization of Thermus thermophilus proline dehydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    White, Tommi A.; Tanner, John J., E-mail: [Departments of Chemistry and Biochemistry, University of Missouri-Columbia, Columbia, Missouri 65211 (United States)


    Cloning, purification and crystallization of T. thermophilus proline dehydrogenase is reported. The detergent n-octyl β-d-glucopyranoside was used to reduce polydispersity, which enabled crystallization. Nature recycles l-proline by converting it to l-glutamate. This four-electron oxidation process is catalyzed by the two enzymes: proline dehydrogenase (PRODH) and Δ{sup 1}-pyrroline-5-carboxylate dehydrogenase. This note reports the cloning, purification and crystallization of Thermus thermophilus PRODH, which is the prototype of a newly discovered superfamily of bacterial monofunctional PRODHs. The results presented here include production of a monodisperse protein solution through use of the detergent n-octyl β-d-glucopyranoside and the growth of native crystals that diffracted to 2.3 Å resolution at Advanced Light Source beamline 4.2.2. The space group is P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 82.2, b = 89.6, c = 94.3 Å. The asymmetric unit is predicted to contain two protein molecules and 46% solvent. Molecular-replacement trials using a fragment of the PRODH domain of the multifunctional Escherichia coli PutA protein as the search model (24% amino-acid sequence identity) did not produce a satisfactory solution. Therefore, the structure of T. thermophilus PRODH will be determined by multiwavelength anomalous dispersion phasing using a selenomethionyl derivative.

  11. Expression and affinity purification of recombinant proteins from plants (United States)

    Desai, Urvee A.; Sur, Gargi; Daunert, Sylvia; Babbitt, Ruth; Li, Qingshun


    With recent advances in plant biotechnology, transgenic plants have been targeted as an inexpensive means for the mass production of proteins for biopharmaceutical and industrial uses. However, the current plant purification techniques lack a generally applicable, economic, large-scale strategy. In this study, we demonstrate the purification of a model protein, beta-glucuronidase (GUS), by employing the protein calmodulin (CaM) as an affinity tag. In the proposed system, CaM is fused to GUS. In the presence of calcium, the calmodulin fusion protein binds specifically to a phenothiazine-modified surface of an affinity column. When calcium is removed with a complexing agent, e.g., EDTA, calmodulin undergoes a conformational change allowing the dissociation of the calmodulin-phenothiazine complex and, therefore, permitting the elution of the GUS-CaM fusion protein. The advantages of this approach are the fast, efficient, and economical isolation of the target protein under mild elution conditions, thus preserving the activity of the target protein. Two types of transformation methods were used in this study, namely, the Agrobacterium-mediated system and the viral-vector-mediated transformation system. Copyright 2002 Elsevier Science (USA).

  12. Simple optimization method for partitioning purification of hydrogen networks

    Directory of Open Access Journals (Sweden)

    W.M. Shehata


    Full Text Available The Egyptian petroleum fuel market is increasing rapidly nowadays. These fuels must be in the standard specifications of the Egyptian General Petroleum Corporation (EGPC, which required lower sulfur gasoline and diesel fuels. So the fuels must be deep hydrotreated which resulted in increasing hydrogen (H2 consumption for deeper hydrotreating. Along with increased H2 consumption for deeper hydrotreating, additional H2 is needed for processing heavier and higher sulfur crude slates especially in hydrocracking process, in addition to hydrotreating unit, isomerization units and lubricant plants. Purification technology is used to increase the amount of recycled hydrogen. If the amount of recycled hydrogen is increased, the amount of hydrogen that is sent to the furnaces with the off gas will decrease. In this work, El Halwagi et al. (2003 and El Halwagi (2012 optimization methods which are used for recycle/reuse integration systems have been extended to be used in the partitioning purification of hydrogen networks to minimize the hydrogen consumption and the hydrogen discharge. An actual case study and two case studies from the literature are solved to illustrate the proposed method.

  13. Travel of pollution, and purification en route, in sandy soils (United States)

    Baars, J. K.


    The travel of pollution in sandy soils, and the extent to which purification takes place en route, are discussed, with special reference to the possible contamination of ground water—a problem which is of particular importance in the Netherlands, where the water-supply for many of the large towns is drawn from the water underneath the dunes. Specifically, two types of soil pollution are considered: (a) severe pollution of the surface layers with matter concentrated in a small volume of water (e.g., faecal matter from pit privies at camping-sites); and (b) moderate pollution of the surface layers with matter contained in large quantities of water (e.g., organic matter and bacteria in river water used for the artificial recharge of ground water). It is shown that in both these types of pollution the self-purification is sufficient to prevent contamination of the ground water, provided that the soil is very fine and—in the case of the first type—dry and well aerated, and provided that the ground-water level is not too high or the rate of infiltration too great. PMID:13472428

  14. Preparation and purification of DNA from insects for AFLP analysis. (United States)

    Reineke, A; Karlovsky, P; Zebitz, C P


    Analysis of amplified fragment length polymorphism (AFLP) has the potential to become a powerful new DNA fingerprinting technique for studying genetic relationships and genetic diversity in arthropods. Since DNA of high quality is a crucial prerequisite for AFLP analysis we evaluated the applicability of six protocols (one fast and four complex methods with phenol-chloroform treatments as well as one CTAB-based method) for extracting DNA from insect material and three additional DNA purification steps. The most rapid DNA isolation method did not produce DNA suitable for AFLP analysis. Among four complex methods tested, two protocols resulted in comparatively low yields of DNA that was therefore not used as template for AFLP analysis. The other two complex methods with phenol treatments and a CTAB-based DNA extraction protocol provided DNA suitable for AFLP assay. An additional purification of the DNA using spermine precipitation revealed a few extra bands in an AFLP gel that were masked in unpurified DNA. Therefore spermine precipitation is recommended for AFLP templates.

  15. Bismuth Oxysulfide and Its Polymer Nanocomposites for Efficient Purification

    Directory of Open Access Journals (Sweden)

    Yidong Luo


    Full Text Available The danger of toxic organic pollutants in both aquatic and air environments calls for high-efficiency purification material. Herein, layered bismuth copper oxychalcogenides, BiCuSO, nanosheets of high photocatalytic activity were introduced to the PVDF (Polyvinylidene Fluoride. The fibrous membranes provide an easy, efficient, and recyclable way to purify organic pollutant. The physical and photophysical properties of the BiCuSO and its polymer composite were characterized by scanning electron microscopy (SEM, X-ray diffraction (XRD, ultraviolet-visible diffuse reflection spectroscopy (DRS, X-ray photoelectron spectroscopy (XPS, electron spin resonance (EPR. Photocatalysis of Congo Red reveals that the BiCuSO/PVDF shows a superior photocatalytic activity of a 55% degradation rate in 70 min at visible light. The high photocatalytic activity is attributed to the exposed active {101} facets and the triple vacant associates V B i ‴ V O • • V B i ‴ . By engineering the intrinsic defects on the surface of bismuth oxysulfide, high solar-driven photocatalytic activity can be approached. The successful fabrication of the bismuth oxysulfide and its polymer nanocomposites provides an easy and general approach for high-performance purification materials for various applications.

  16. Purification of Methyl Ricinoleate on Producing of Cetane Improver (United States)

    Abdullah; Triyono; Trisunaryanti, W.; Haryadi, W.


    Purification of methyl ricinoleate on a mixture of methyl ester that was derived from castor oil has been conducted. Methyl ricinoleate with high purity is required in the manufacture of additives for improving the cetane number of diesel oil. Purification of methyl ricinoleate was performed by fractional distillation under reduced pressure at 4.5 mmHg. In order to produce of cetane improver, methyl ricinoleate was nitrated with a mixture of HNO3 and acetic anhydride at 11-13°C for 4 hours. Based on the analysis using gas chromatography (GC) was known that purity of methyl ricinoleate can be increased from 87.2% to 99.5%, with a yield of 30.6% (v/v). FTIR analysis showedthe absorption spectrum at 3440 (1 /cm), which is derived from the vibrations of the -OH (hydroxyl) on the ricinoleate methyl molecules. The results of characterization showed an increase in density of 0.9161 (mg/L) to 0.9171 (mg /L) and the refractive index of 1.4619 to 1.4627. FTIR spectra of nitrated methyl ricinoleateshowedan absorption spectrum at 1627 (1/cm), in which indicates a new compound with nitrate group (-NO3).

  17. The Expanding Toolkit of Translating Ribosome Affinity Purification. (United States)

    Dougherty, Joseph D


    Translating ribosome affinity purification is a method initially developed for profiling mRNA from genetically defined cell types in complex tissues. It has been applied both to identify target molecules in cell types that are important for controlling a variety of behaviors in the brain, and to understand the molecular consequences on those cells due to experimental manipulations, ranging from drugs of abuse to disease-causing mutations. Since its inception, a variety of methodological advances are opening new avenues of investigation. These advances include a variety of new methods for targeting cells for translating ribosome affinity purification by features such as their projections or activity, additional tags and mouse reagents increasing the flexibility of the system, and new modifications of the method specifically focused on studying the regulation of translation. The latter includes methods to assess cell type-specific regulation of translation in specific subcellular compartments. Here, I provide a summary of these recent advances and resources, highlighting both new experimental opportunities and areas for future technical development. Copyright © 2017 the authors 0270-6474/17/3712079-09$15.00/0.

  18. Production and purification of polyclonal antibody against melatonin hormone

    Directory of Open Access Journals (Sweden)

    Fooladsaz K


    Full Text Available Nowadays immunochemical techniques have played a very important and valuable role in quantitative and qualitative assays of liquid compounds of the body. Producing antibody against immunogenes is the first step to make immunochemical kits. In this study production and purification of polyclonal antibody against melatonin has been considered. This hormone which has several important functions in physiological conditions such as migraine, cirrhosis, mammary gland cancer and other diseases, is the most important pineal gland secretion. This gland is a circumventricular organ of brain and according to histological and anatomical studies, it is a high secretory organ, that secretes active biological substances like melatonin, oxytocin, serotonin and ect. In this study, melatonin has been considered as hapten and has become an immunogen by being linked to the bovine serum Albumin. Then, by the immunization of three white New Zeland rabbits that had the booster injections in regular intervals, the antibody titer was detected to be 1/2000, by using checkboard curves, and with the use of melatonin linked to penicillinase as a labeled antigen, the titer was detected 1/200. Finally an antibody with high purification rate has been obtained, which can be used in immunochemical assays like RIA, ELISA, and EIA.

  19. Design of laboratory cyclone separator for biogas purification

    Directory of Open Access Journals (Sweden)

    Marián Fodora


    Full Text Available This article deals with calculation of a cyclone separator for biogas purification using physical and chemical methods. There is presented a methodology for determination of operating dimensions of the cyclone separator and description of principal features of the cyclone separator model. Calculations have been performed for the diameter of the cylindrical part of cyclone separator 175 mm and for the biogas volume flow rate 6.9∙10−5 m3∙s−1. The calculations can be used in practice for the design of cyclone separator depending on the flow rate of biogas, size of the biogas plants respectively. The developed cyclone separator has been used for the cleaning of biogas in operating conditions at the biogas plant in Kolinany (Slovakia. The presented method of biogas purification has been used for the removing of hydrogen sulphide, particulate matter and carbon dioxide from the raw biogas at the biogas plant. Removal of these undesirable impurities from the biogas is an important step in the production of a fully valued fuel, biomethane.

  20. Purification of novel peptide antibiotics from human milk. (United States)

    Liepke, C; Zucht, H D; Forssmann, W G; Ständker, L


    A strategy was established for the identification of novel antimicrobial peptides from human milk. For the generation of bioactive peptides human milk was acidified and proteolyzed with pepsin simulating the digest in infants stomachs. Separation of proteins and resulting fragments was performed by means of reversed-phase chromatography detecting the antimicrobial activity of each fraction using a sensitive radial diffusion assay. In order to avoid the purification of the known abundant antimicrobial milk protein lysozyme, it was identified in HPLC fractions by its enzymatic activity and by matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS). On condition that lysozyme was not detectable and antibacterial activity of HPLC fractions was caused by a peptide, which was confirmed by proteolytic cleavage leading to a loss of activity, further purification was performed by consecutive chromatographic steps guided by the antibacterial assay. Using this strategy, an as yet unknown casein fragment exhibiting antimicrobial activity was purified in addition to antimicrobial lactoferrin fragments. The new antimicrobial peptide resembles a proteolytic fragment of human casein-K (residues 63-117) and inhibits the growth of gram-positive, gram-negative bacteria, and yeasts. Our results confirm that antimicrobially-active peptides are liberated from human milk proteins during proteolytic hydrolysis and may play an important role in the host defense system of the newborn.

  1. Purification of sodium phosphate yielded from Bangka's monazite base decomposition

    International Nuclear Information System (INIS)

    Walujo, Sugeng; Susilaningtyas; Mukhlis; Tukardi


    The aim of this experiment is to get purification condition of sodium phosphate from the filtration result of mixing mother liquor and filtrate of washing residue from Bangka monazite decomposition by alkaline. The method of purification which has been used was: dissolved the precipitation of sodium phosphate into water with the agitation time constant at 5 minutes and then the solution is settled for 12 hours until the sodium phosphate crystals appear. The variable of experiment included of dissolution time and the ratio of the amount precipitate sodium phosphate, which dissolved against the volume of water as solvent. Experiment data shown that the good temperature of dissolution at 70 oC with the ratio of precipitate sodium phosphate which dissolved is 60 gram/100 ml of water. The recovery of sodium phosphate crystallization is 65.18 % with Na 3 PO 4 purity is about 65.608 %, and it impurities content of U is 0.007% and NaOH and the others are 34.383%

  2. Specificities of micro-reactors for hydrogen production and purification

    Energy Technology Data Exchange (ETDEWEB)

    Mirodatos, C.; Dupont, N.; Germani, G.; Veen, A. C. ven; Schuurman, Y.


    Sustainable chemistry and exploitation of energy sources for the next decades requires considerable progress in process intensification. A development of new tools and equipments meeting the objectives of high efficiency, improved safety, compactness and low implementation costs is therefore subject of intensive research effort. Among the various scenarios tested in R and D, micro-structured reactors appear as a highly promising technology 1 and perspectives of mass production are already announced by technology providers 2. These reactors are based on assembly/stacking of micro structured plates or fibres. Due to their high heat and/or mass transfer, low pressure drop and good phase contacting, they sound particularly adapted to the large domain of hydrogen production by fuel reforming and purification. This presentation aims at outlining the state of the art, the advantages and drawbacks of using micro-structured reactors to intensify hydrogen production and purification. Two case studies will illustrate this approach: i) comparison between fixed bed and micro-structured reactor for the reforming of methanol into hydrogen and carbon oxides and ii) use of those devices in kinetic studies on the WGS reaction. (Author)

  3. Purification and characterization of a thylakoid protein kinase

    International Nuclear Information System (INIS)

    Coughlan, S.J.; Hind, G.


    Control of state transitions in the thylakoid by reversible phosphorylation of the light-harvesting chlorophyll a/b protein complex of photosystem II (LHC-II) is modulated by a kinase. The kinase catalyzing this phosphorylation is associated with the thylakoid membrane, and is regulated by the redox state of the plastoquinone pool. The isolation and partial purification from spinach thylakoids of two protein kinases (CPK1, CPK2) of apparent molecular masses 25 kDa and 38 kDa has been reported. Neither enzyme utilizes isolated LHC-II as a substrate. The partial purification of a third protein kinase (LHCK) which can utilize both lysine-rich histones (IIIs and Vs) and isolated LHC-II as substrate has now been purified to homogeneity and characterized by SDS-polyacrylamide gel electrophoresis as a 64 kDa peptide. From a comparison of the two isolation procedures we have concluded that CPK1 is indeed a protein kinase, but has a lower specific activity than that of LHCK. 8 refs., 4 figs

  4. Cloning, purification and crystallization of Thermus thermophilus proline dehydrogenase

    International Nuclear Information System (INIS)

    White, Tommi A.; Tanner, John J.


    Cloning, purification and crystallization of T. thermophilus proline dehydrogenase is reported. The detergent n-octyl β-d-glucopyranoside was used to reduce polydispersity, which enabled crystallization. Nature recycles l-proline by converting it to l-glutamate. This four-electron oxidation process is catalyzed by the two enzymes: proline dehydrogenase (PRODH) and Δ 1 -pyrroline-5-carboxylate dehydrogenase. This note reports the cloning, purification and crystallization of Thermus thermophilus PRODH, which is the prototype of a newly discovered superfamily of bacterial monofunctional PRODHs. The results presented here include production of a monodisperse protein solution through use of the detergent n-octyl β-d-glucopyranoside and the growth of native crystals that diffracted to 2.3 Å resolution at Advanced Light Source beamline 4.2.2. The space group is P2 1 2 1 2 1 , with unit-cell parameters a = 82.2, b = 89.6, c = 94.3 Å. The asymmetric unit is predicted to contain two protein molecules and 46% solvent. Molecular-replacement trials using a fragment of the PRODH domain of the multifunctional Escherichia coli PutA protein as the search model (24% amino-acid sequence identity) did not produce a satisfactory solution. Therefore, the structure of T. thermophilus PRODH will be determined by multiwavelength anomalous dispersion phasing using a selenomethionyl derivative


    Directory of Open Access Journals (Sweden)

    R. Satheeskumar


    Full Text Available Partial purification and characterization of alkalophilic protease production from Pseudomonas aeruginosa was isolated from the gut of marine and coastal waters shrimp Penaeus monodon. The protease production was assayed in submerged fermentation to produce maximum protease activity (423 ± 0.09 U/ml. The enzyme was precipitated with ammonium sulphate and partially purified by ion exchange chromatography through DEAE Sephadex A-50 column. In 10th fraction showed maximum protease activity (734 ± 0.18 U/ml with increase in purification fold. The molecular weight of protease from Pseudomonas aeruginosa was recorded as 60 kDa. The stability of protease was tested at various pH and temperature; it showed maximum protease activity at pH-9 and temperature 50ºC. Among the various surfactants tested for enzyme stability, maximum activity was retained in poly ethylene glycol. The compatibility of protease enzyme with various commercial detergents; the enzyme retained maximum protease activity in tide. The results are indicated that all these properties make the bacterial proteases are most suitable for wide industrial applications.

  6. Concentration-Purification of Uranium from an Acid Leaching Solution (United States)

    Guettaf, H.; Becis, A.; Ferhat, K.; Hanou, K.; Bouchiha, D.; Yakoubi, K.; Ferrad, F.


    Chemical processes for the elaboration of uranium concentrate from uranium ore have been studied. This process is composed of successive units operations: crushing, milling, acid conventional leaching, filtration-washing, purification-concentration by ion exchange resins and uranium precipitation. The acid leaching operating conditions allow us to obtain a recovery uranium rate of 93%. The uranium concentration of the pregnant solution is approximately of 1.2 g/l. This value justifies the use of ion exchange resins to the concentration-purification of our pregnant solution. We have noticed that the pregnant solution contains a relatively high phosphate concentration which causes a premature uranium precipitation at pH=1.8. This pH value is in general, considered optimal to obtain the highest amount of fixed uranium by the anionic resin. To avoid the precipitation of uranium, the pH=1.5 has been fixed. We have obtained at this condition a good adsorption capacity. A 75% uranium concentrate have been elaborated, but the filtration of this concentrate has been very difficult. We have also noticed an excessive sulphate concentration. In order to improve this process, we have tested nitrates as eluant at different operating conditions.

  7. [Improved protein-A chromatography for monoclonal antibody purification]. (United States)

    Chen, Quan; Toh, Phyllicia; Hoi, Aina; Xian, Mo; Peng, Xinying; Yang, Yuansheng; Zhang, Haibo; Nian, Rui; Zhang, Wei


    Therapeutic monoclonal antibodies become the major product class within the biopharmaceutical market. Protein A as the first capture step is still dominant in current platforms for purification of monoclonal antibodies. In this study, we developed a new antibody harvest process that incorporates acidic treatment of cell harvest, demonstrating high process yield, improved clearance of host cell associated contaminants, like non-histone host cell protein, histone, DNA and heteroaggregates. Host protein contamination was reduced about 10-fold compared to protein A loaded with harvest clarified by centrifugation and microfiltration. Turbidity increase of eluted IgG upon pH neutralization was nearly eliminated. Residual levels of impurities in the protein A eluate were achieved that potentially meet requirements of drug substance and thus alleviate the burden for further impurities removal in subsequent chromatography steps. The mechanism of host cell associated contaminants removal during acidic treatment was also explored. After a polishing step by Capto adhere, host cell protein was reduced to less than 5 ppm, DNA less than 1 ppb, histone to undetectable level, heteroaggregates less than 0.01% with total IgG recovery around 87%. This efficient process can be easily integrated into current IgG purification platforms, and may overcome downstream processing challenges.

  8. Expression, purification and crystallization of a lyssavirus matrix (M) protein

    Energy Technology Data Exchange (ETDEWEB)

    Assenberg, René [Division of Structural Biology and Oxford Protein Production Facility, The Henry Wellcome Building for Genomic Medicine, Oxford University, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Delmas, Olivier [UPRE Lyssavirus Dynamics and Host Adaptation, WHO Collaborating Centre for Reference and Research on Rabies, Institut Pasteur, 28 Rue du Docteur Roux, 75724 Paris CEDEX 15 (France); Graham, Stephen C.; Verma, Anil; Berrow, Nick; Stuart, David I.; Owens, Raymond J. [Division of Structural Biology and Oxford Protein Production Facility, The Henry Wellcome Building for Genomic Medicine, Oxford University, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Bourhy, Hervé [UPRE Lyssavirus Dynamics and Host Adaptation, WHO Collaborating Centre for Reference and Research on Rabies, Institut Pasteur, 28 Rue du Docteur Roux, 75724 Paris CEDEX 15 (France); Grimes, Jonathan M., E-mail: [Division of Structural Biology and Oxford Protein Production Facility, The Henry Wellcome Building for Genomic Medicine, Oxford University, Roosevelt Drive, Oxford OX3 7BN (United Kingdom)


    The expression, purification and crystallization of the full-length matrix protein from three lyssaviruses is described. The matrix (M) proteins of lyssaviruses (family Rhabdoviridae) are crucial to viral morphogenesis as well as in modulating replication and transcription of the viral genome. To date, no high-resolution structural information has been obtained for full-length rhabdovirus M. Here, the cloning, expression and purification of the matrix proteins from three lyssaviruses, Lagos bat virus (LAG), Mokola virus and Thailand dog virus, are described. Crystals have been obtained for the full-length M protein from Lagos bat virus (LAG M). Successful crystallization depended on a number of factors, in particular the addition of an N-terminal SUMO fusion tag to increase protein solubility. Diffraction data have been recorded from crystals of native and selenomethionine-labelled LAG M to 2.75 and 3.0 Å resolution, respectively. Preliminary analysis indicates that these crystals belong to space group P6{sub 1}22 or P6{sub 5}22, with unit-cell parameters a = b = 56.9–57.2, c = 187.9–188.6 Å, consistent with the presence of one molecule per asymmetric unit, and structure determination is currently in progress.

  9. Purification of bacteriophage M13 by anion exchange chromatography. (United States)

    Monjezi, Razieh; Tey, Beng Ti; Sieo, Chin Chin; Tan, Wen Siang


    M13 is a non-lytic filamentous bacteriophage (phage). It has been used widely in phage display technology for displaying foreign peptides, and also for studying macromolecule structures and interactions. Traditionally, this phage has been purified by cesium chloride (CsCl) density gradient ultracentrifugation which is highly laborious and time consuming. In the present study, a simple, rapid and efficient method for the purification of M13 based on anion exchange chromatography was established. A pre-packed SepFast Super Q column connected to a fast protein liquid chromatography (FPLC) system was employed to capture released phages in clarified Escherichia coli fermented broth. An average yield of 74% was obtained from a packed bed mode elution using citrate buffer (pH 4), containing 1.5 M NaCl at 1 ml/min flow rate. The purification process was shortened substantially to less than 2 h from 18 h in the conventional ultracentrifugation method. SDS-PAGE revealed that the purity of particles was comparable to that of CsCl gradient density ultracentrifugation method. Plaque forming assay showed that the purified phages were still infectious. Copyright 2010 Elsevier B.V. All rights reserved.

  10. New research on bioregenerative air/water purification systems (United States)

    Johnson, Anne H.; Ellender, R. D.; Watkins, Paul J.


    For the past several years, air and water purification systems have been developed and used. This technology is based on the combined activities of plants and microorganisms as they function in a natural environment. More recently, researchers have begun to address the problems associated with indoor air pollution. Various common houseplants are currently being evaluated for their abilities to reduce concentrations of volatile organic compounds (VOCS) such as formaldehyde and benzene. With development of the Space Exploration Initiative, missions will increase in duration, and problems with resupply necessitates implementation of regenerative technology. Aspects of bioregenerative technology have been included in a habitat known as the BioHome. The ultimate goal is to use this technology in conjunction with physicochemical systems for air and water purification within closed systems. This study continued the risk assessment of bioregenerative technology with emphasis on biological hazards. In an effort to evaluate the risk for human infection, analyses were directed at enumeration of fecal streptococci and enteric viruses with the BioHome waste water treatment system.

  11. Multimodal charge-induction chromatography for antibody purification. (United States)

    Tong, Hong-Fei; Lin, Dong-Qiang; Chu, Wen-Ning; Zhang, Qi-Lei; Gao, Dong; Wang, Rong-Zhu; Yao, Shan-Jing


    Hydrophobic charge-induction chromatography (HCIC) has advantages of high capacity, salt-tolerance and convenient pH-controlled elution. However, the binding specificity might be improved with multimodal molecular interactions. New ligand W-ABI that combining tryptophan and 5-amino-benzimidazole was designed with the concept of mutimodal charge-induction chromatography (MCIC). The indole and benzimidazole groups of the ligand could provide orientated mutimodal binding to target IgG under neutral pH, while the imidazole groups could induce the electrostatic repulsion forces for efficient elution under acidic pH. W-ABI ligand was coupled successfully onto agarose gel, and IgG adsorption behaviors were investigated. High affinity to IgG was found with the saturated adsorption capacity of 70.4 mg/ml at pH 7, and the flow rate of mobile phase showed little impact on the dynamic binding capacity. In addition, efficient elution could be achieved at mild acidic pH with high recovery. Two separation cases (IgG separation from albumin containing feedstock and monoclonal antibody purification from cell culture supernatant) were verified with high purity and recovery. In general, MCIC with the specially-designed ligand is an expanding of HCIC with improved adsorption selectivity, which would be a potential alternative to Protein A-based capture for the cost-effective purification of antibodies. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Purification of biodiesel by choline chloride based deep eutectic solvent (United States)

    Niawanti, Helda; Zullaikah, Siti; Rachimoellah, M.


    Purification is a crucial step in biodiesel production to meet the biodiesel standard. This study purified biodiesel using choline chloride based deep eutectic solvent (DES). DES was used to reduce unreacted oil and unsaponifiable matter in rice bran oil based biodiesel. The objective of this work was to study the effect of extraction time using DES on the content and yield of fatty acid methyl ester (FAME). Rice bran used in this work contains 16.49 % of oil with initial free fatty acids (FFA) of 44.75 %. Acid catalyzed methanolysis was employed to convert rice bran oil (RBO) into biodiesel under following operation conditions: T = 60 °C, t = 8 h, molar ratio of oil to methanol = 1/10, H2SO4 = 1% w/w of oil. Rice bran oil based biodiesel obtained contain 89.05 % of FAME with very low FFA content (0.05 %). DES was made from a mixture of choline chloride and ethylene glycol with molar ratio of 1/2. Molar ratio of crude biodiesel to DES were 1/2 and 1/4. Extraction time was varied from 15 minutes to 240 minutes at 30 °C. The highest FAME content was obtained after purification for 240 min. at molar ratio crude biodiesel to DES 1/4 was 96.60 %. This work shows that DES has potential to purify biodiesel from non-edible raw material, such as RBO.

  13. Production and Purification of Peroxidase from Aspergillus niger.

    Directory of Open Access Journals (Sweden)

    Mohammed A. Jebor


    Full Text Available This study was conducted in the laboratories of Biology Department, College of Science, which deals with isolation and purification of peroxidase and optimization of process parameters to achieve maximum yield of peroxidase by Aspergillus niger. Solid-state fermentation of Aspergillus niger was carried out for enhanced production of peroxidase using hydrogen peroxide as the substrate of enzyme maximum activity of the enzyme was achieved under optimum growth conditions. The optimum conditions were the isolated of Aspergillus niger from soil and growth in synthetic medium, it gave high titer of peroxidase activity, the fructose as carbon source, peptone as nitrogen source, after 12 days of incubation, incubation temperature 25 °C and pH = 6.5. Peroxidase purified in four purification steps; precipitation with 70% saturation of ammonium sulfate, step of dialysis, the third by ion exchange chromatography using DEAE-Cellulose and fourth by gel filtration throughout Sephadex G-100. The specific activity of the purified enzyme was 150U/mg with 7.75 folds. The peroxidase was shown to have molecular weight of 40kDa in SDS-PAGA and about 40kDa in gel filtration.The optimum pH and temperature for peroxidase activity 7 and 35 C0 respectively.

  14. Isolation and Purification of Jacalin from Artocarpus Heterophyllus Lam

    Directory of Open Access Journals (Sweden)

    M.R. Othman


    Full Text Available This paper presents investigation results of saturation conditions needed for purification of jacalin lectin from the extract seeds of Artocarpus heterophyllus by ammonium precipitation and affinity chromatography on Galactose-Affi gel Hz. Three different aspects of parameters encompassing the percentage of saturation of ammonium sulfate precipitation, the presence of ammonium sulfate on Lowry method and the suitable galactose concentration for optimum elution of the protein from Galactose-Affi gel Hz were investigated. With three different sets of fractional saturation of jacalin purification using ammonium sulfate precipitation, the maximum yield of 0.463 g/g was achieved at 0-90% saturation range in the absence of dialysis. Maximum yield of 0.425 g/g was obtained at 30-60% and 0-90% saturation range in the presence of dialysis. The result from this work also indicates that excessive quantity of NH4SO4 interferes with Lowry method for protein determination substantially. The 0-90% saturation range was found to be more potentially appropriate for large scale application than 30-60% saturation, since the former involves only 1 step NH4SO4 addition. From the affinity chromatography, elution of 0.2 M galactose (in 0.15 M NaCl from Galactose-Affi gel Hz produced the maximum peak profile and jacalin concentration. A reduction or increase in galactose concentration of more than 0.2 M did not increase concentration of purified jacalin purified using this method.

  15. Genotypic Characterization of Azotobacteria Isolated from Argentinean Soils and Plant-Growth-Promoting Traits of Selected Strains with Prospects for Biofertilizer Production

    Directory of Open Access Journals (Sweden)

    Esteban Julián Rubio


    Full Text Available The genetic diversity among 31 putative Azotobacter isolates obtained from agricultural and non-agricultural soils was assessed using rep-PCR genomic fingerprinting and identified to species level by ARDRA and partial 16S rRNA gene sequence analysis. High diversity was found among the isolates, identified as A. chroococcum, A. salinestris, and A. armeniacus. Selected isolates were characterized on the basis of phytohormone biosynthesis, nitrogenase activity, siderophore production, and phosphate solubilization. Indole-3 acetic-acid (IAA, gibberellin (GA3 and zeatin (Z biosynthesis, nitrogenase activity, and siderophore production were found in all evaluated strains, with variation among them, but no phosphate solubilization was detected. Phytohormones excreted to the culture medium ranged in the following concentrations: 2.2–18.2 μg IAA mL−1, 0.3–0.7 μg GA3 mL−1, and 0.5–1.2 μg Z mL−1. Seed inoculations with further selected Azotobacter strains and treatments with their cell-free cultures increased the number of seminal roots and root hairs in wheat seedlings. This latter effect was mimicked by treatments with IAA-pure solutions, but it was not related to bacterial root colonization. Our survey constitutes a first approach to the knowledge of Azotobacter species inhabiting Argentinean soils in three contrasting geographical regions. Moreover, this phenotypic characterization constitutes an important contribution to the selection of Azotobacter strains for biofertilizer formulations.

  16. Genotypic characterization of Azotobacteria isolated from Argentinean soils and plant-growth-promoting traits of selected strains with prospects for biofertilizer production. (United States)

    Rubio, Esteban Julián; Montecchia, Marcela Susana; Tosi, Micaela; Cassán, Fabricio Darío; Perticari, Alejandro; Correa, Olga Susana


    The genetic diversity among 31 putative Azotobacter isolates obtained from agricultural and non-agricultural soils was assessed using rep-PCR genomic fingerprinting and identified to species level by ARDRA and partial 16S rRNA gene sequence analysis. High diversity was found among the isolates, identified as A. chroococcum, A. salinestris, and A. armeniacus. Selected isolates were characterized on the basis of phytohormone biosynthesis, nitrogenase activity, siderophore production, and phosphate solubilization. Indole-3 acetic-acid (IAA), gibberellin (GA3) and zeatin (Z) biosynthesis, nitrogenase activity, and siderophore production were found in all evaluated strains, with variation among them, but no phosphate solubilization was detected. Phytohormones excreted to the culture medium ranged in the following concentrations: 2.2-18.2 μ g IAA mL(-1), 0.3-0.7 μ g GA3 mL(-1), and 0.5-1.2 μ g Z mL(-1). Seed inoculations with further selected Azotobacter strains and treatments with their cell-free cultures increased the number of seminal roots and root hairs in wheat seedlings. This latter effect was mimicked by treatments with IAA-pure solutions, but it was not related to bacterial root colonization. Our survey constitutes a first approach to the knowledge of Azotobacter species inhabiting Argentinean soils in three contrasting geographical regions. Moreover, this phenotypic characterization constitutes an important contribution to the selection of Azotobacter strains for biofertilizer formulations.

  17. Plant growth promoting characterization of indigenous Azotobacteria isolated from soils in Iran. (United States)

    Farajzadeh, Davoud; Yakhchali, Bagher; Aliasgharzad, Naser; Sokhandan-Bashir, Nemat; Farajzadeh, Malak


    It has been well known that the bacteria of the genus Azotobacter, in addition to the beneficial N(2)-fixing activity, are able to improve plant growth by a number of direct and indirect mechanisms. To identify this potential in indigenous azotobacteria, the efficiency of 17 isolates of Azotobacter from the rhizosphere of wheat and barley plants cultivated in salt- and/or drought-affected soils in Iran were evaluated for their ability to dissolve inorganic and organic phosphates, siderophore secretion, indole acetic acid (IAA) production; and protease, chitinase, and ACC deaminase (ACCD) activities. First, they were biochemically characterized and one isolate (strain) was identified by 16S rDNA sequencing. Eight isolates were designated as Azotobacter vinelandii and the remaining isolates were identified as A. chroococcum. All isolates hydrolyzed the organic and inorganic phosphate compounds and effectively produced IAA. Fifteen isolates produced siderophore, but only one isolate showed protease activity which is being reported for the first time in relation to Azotobacter. None of the 17 isolates was capable of producing ACCD or chitinase. However, polymerase chain reaction amplification of the ACCD coding genes, by the use of the gene-specific primers, indicated that not all contain the ACCD gene. The standard screening methods with slight modifications, especially in the case of ACCD assay, were applied. The results showed that the use of specific screening methods, modified according to bacterial nutritional requirements, are the efficient methods for precise evaluation of the plant growth promoting rhizobacteria activity.

  18. Purification of crude glycerol from transesterification reaction of palm oil using direct method and multistep method (United States)

    Nasir, N. F.; Mirus, M. F.; Ismail, M.


    Crude glycerol which produced from transesterification reaction has limited usage if it does not undergo purification process. It also contains excess methanol, catalyst and soap. Conventionally, purification method of the crude glycerol involves high cost and complex processes. This study aimed to determine the effects of using different purification methods which are direct method (comprises of ion exchange and methanol removal steps) and multistep method (comprises of neutralization, filtration, ion exchange and methanol removal steps). Two crude glycerol samples were investigated; the self-produced sample through the transesterification process of palm oil and the sample obtained from biodiesel plant. Samples were analysed using Fourier Transform Infrared Spectroscopy, Gas Chromatography and High Performance Liquid Chromatography. The results of this study for both samples after purification have showed that the pure glycerol was successfully produced and fatty acid salts were eliminated. Also, the results indicated the absence of methanol in both samples after purification process. In short, the combination of 4 purification steps has contributed to a higher quality of glycerol. Multistep purification method gave a better result compared to the direct method as neutralization and filtration steps helped in removing most excess salt, fatty acid and catalyst.

  19. Waste water biological purification plants of dairy products industry and energy management (United States)

    Stepanov, Sergey; Solkina, Olga; Stepanov, Alexander; Zhukova, Maria


    The paper presents results of engineering and economical comparison of waste water biological purification plants of dairy products industry. Three methods of purification are compared: traditional biological purification with the use of secondary clarifiers and afterpurification through granular-bed filters, biomembrane technology and physical-and-chemical treatment together with biomembrane technology for new construction conditions. The improvement of the biological purification technology using nitro-denitrification and membrane un-mixing of sludge mixture is a promising trend in this area. In these calculations, an energy management which is widely applied abroad was used. The descriptions of the three methods are illustrated with structural schemes. Costs of equipment and production areas are taken from manufacturers’ data. The research is aimed at an engineering and economical comparison of new constructions of waste water purification of dairy products industry. The experiment demonstrates advantages of biomembrane technology in waste water purification. This technology offers prospects of 122 million rubles cost saving during 25 years of operation when compared with of the technology of preparatory reagent flotation and of 13.7 million rubles cost saving compared to the option of traditional biological purification.

  20. The Protein Maker: an automated system for high-throughput parallel purification

    International Nuclear Information System (INIS)

    Smith, Eric R.; Begley, Darren W.; Anderson, Vanessa; Raymond, Amy C.; Haffner, Taryn E.; Robinson, John I.; Edwards, Thomas E.; Duncan, Natalie; Gerdts, Cory J.; Mixon, Mark B.; Nollert, Peter; Staker, Bart L.; Stewart, Lance J.


    The Protein Maker instrument addresses a critical bottleneck in structural genomics by allowing automated purification and buffer testing of multiple protein targets in parallel with a single instrument. Here, the use of this instrument to (i) purify multiple influenza-virus proteins in parallel for crystallization trials and (ii) identify optimal lysis-buffer conditions prior to large-scale protein purification is described. The Protein Maker is an automated purification system developed by Emerald BioSystems for high-throughput parallel purification of proteins and antibodies. This instrument allows multiple load, wash and elution buffers to be used in parallel along independent lines for up to 24 individual samples. To demonstrate its utility, its use in the purification of five recombinant PB2 C-terminal domains from various subtypes of the influenza A virus is described. Three of these constructs crystallized and one diffracted X-rays to sufficient resolution for structure determination and deposition in the Protein Data Bank. Methods for screening lysis buffers for a cytochrome P450 from a pathogenic fungus prior to upscaling expression and purification are also described. The Protein Maker has become a valuable asset within the Seattle Structural Genomics Center for Infectious Disease (SSGCID) and hence is a potentially valuable tool for a variety of high-throughput protein-purification applications

  1. Preparation and purification of organic samples for selenium isotope studies. (United States)

    Banning, Helena; Stelling, Monika; König, Stephan; Schoenberg, Ronny; Neumann, Thomas


    Selenium (Se) is an important micronutrient but also a strong toxin with a narrow tolerance range for many organisms. As such, a globally heterogeneous Se distribution in soils is responsible for various disease patterns (i.e. Se excess and deficiency) and environmental problems, whereby plants play a key role for the Se entrance into the biosphere. Selenium isotope variations were proved to be a powerful tracer for redox processes and are therefore promising for the exploration of the species dependent Se metabolism in plants and the Se cycling within the Critical Zone. Plant cultivation setups enable systematic controlled investigations, but samples derived from them-plant tissue and phytoagar-are particularly challenging and require specific preparation and purification steps to ensure precise and valid Se isotope analytics performed with HG-MC-ICP-MS. In this study, different methods for the entire process from solid tissue preparation to Se isotope measurements were tested, optimized and validated. A particular microwave digestion procedure for plant tissue and a vacuum filtration method for phytoagar led to full Se recoveries, whereby unfavorable organic residues were reduced to a minimum. Three purification methods predominantly described in the literature were systematically tested with pure Se solution, high concentrated multi-element standard solution as well as plant and phytoagar as target matrices. All these methods efficiently remove critical matrix elements, but differ in Se recovery and organic residues. Validation tests doping Se-free plant material and phytoagar with a reference material of known Se isotope composition revealed the high impact of organic residues on the accuracy of MC-ICP-MS measurements. Only the purification method with no detectable organic residues, hydride generation and trapping, results in valid mass bias correction for plant samples with an average deviation to true δ82/76Se values of 0.2 ‰ and a reproducibility (2 SD

  2. 123I and 13I purification for biomolecules labelling

    International Nuclear Information System (INIS)

    Catanoso, Marcela Forli


    The 123 I and 131 I are iodine radioisotopes widely used in Nuclear Medicine. The radioisotope 123 I is used in diagnosis through the SPECT technique and is routinely produced at IPEN in cyclotron through the reaction: '1 24 Xe (p, 2n) '1 23 Cs -> 123 Xe -> 123 I. The radioisotope 131 I is used both in diagnosis and therapy due to its physical characteristics of decay by β - and its γ-ray emissions that are softened with the use of specific collimators for diagnosis. It is routinely produced at IPEN using the nuclear reactor through the indirect reaction: 130 Te (n, γ) -> 131 Te -> 131 I, irradiating compounds containing Te. The radiopharmaceuticals prepared with these radioisotopes go through rigorous quality control tests and the chemical purity of the primary radioisotopes 123 I and 131 I are within the permissible limits currently defined. However, the presence of some chemical contaminants can prejudice the biomolecules labeling (monoclonal antibodies and peptides), that will produce radiopharmaceuticals of first generation to the oncology area. The aim of this work was to obtain a new purification method of these radioisotopes, allowing the labeling of biomolecules and also to established a process control on those radioisotopes. The methodology was separated on 3 steps: Evaluation of '1 23 I e 131 I radionuclidic purity using a hyper pure germanium detector, chemical purity using ICP-OES and the retention and elution study of 131 I in several absorbers to choose the most appropriate for the purification tests analyzing the behavior of the possible contaminants. The radionuclidic analyses showed the presence of Te and Co on 131 I samples and Te, Tc e Co on 123 I samples. The chemical purity analyses showed the presence of Al and Mo in 123 I, coming from the window material of the target holder and the presence of Al and Te in 131 I samples, coming from the target holder and the target, respectively. The retention and elution study selected the most

  3. Total chemical synthesis of proteins without HPLC purification. (United States)

    Loibl, S F; Harpaz, Z; Zitterbart, R; Seitz, O


    The total chemical synthesis of proteins is a tedious and time-consuming endeavour. The typical steps involve solid phase synthesis of peptide thioesters and cysteinyl peptides, native chemical ligation (NCL) in solution, desulfurization or removal of ligation auxiliaries in the case of extended NCL as well as many intermediary and final HPLC purification steps. With an aim to facilitate and improve the throughput of protein synthesis we developed the first method for the rapid chemical total on-resin synthesis of proteins that proceeds without a single HPLC-purification step. The method relies on the combination of three orthogonal protein tags that allow sequential immobilization ( via the N-terminal and C-terminal ends), extended native chemical ligation and release reactions. The peptide fragments to be ligated are prepared by conventional solid phase synthesis and used as crude materials in the subsequent steps. An N-terminal His 6 unit permits selective immobilization of the full length peptide thioester onto Ni-NTA agarose beads. The C-terminal peptide fragment carries a C-terminal peptide hydrazide and an N-terminal 2-mercapto-2-phenyl-ethyl ligation auxiliary, which serves as a reactivity tag for the full length peptide. As a result, only full length peptides, not truncation products, react in the subsequent on-bead extended NCL. After auxiliary removal the ligation product is liberated into solution upon treatment with mild acid, and is concomitantly captured by an aldehyde-modified resin. This step allows the removal of the most frequently observed by-product in NCL chemistry, i.e. the hydrolysed peptide thioester (which does not contain a C-terminal peptide hydrazide). Finally, the target protein is released with diluted hydrazine or acid. We applied the method in the synthesis of 46 to 126 amino acid long MUC1 proteins comprising 2-6 copies of a 20mer tandem repeat sequence. Only three days were required for the parallel synthesis of 9 MUC1 proteins

  4. A metodologia de superfície de resposta como ferramenta para a avaliação da produção de alginato e poli-hidroxibutirato pela Azotobacter vinelandii - doi: 10.4025/actascitechnol.v32i2.1792

    Directory of Open Access Journals (Sweden)

    Adriana Navarro da Silva


    Full Text Available O alginato é um polissacarídeo normalmente extraído de paredes celulares de algas marrons e utilizado nas indústrias de alimentos, farmacêuticas e biotecnológicas. A produção é concentrada no cultivo de algas marinhas marrons, mas várias bactérias do gênero Pseudomonas e Azotobacter produzem alginato. A estrutura química dos alginatos produzidos por algas é similar a dos sintetizados pela A. vinelandii. Esta bactéria também produz polímeros intracelulares como o poli-hidroxibutirato (PHB, conhecido como bioplástico. Neste trabalho, estudou-se a produção simultânea do alginato e PHB pela A. vinelandii, utilizando-se sacarose e diferentes parâmetros de fermentação em agitador orbital rotatório. Os valores ótimos para produção destes compostos foram determinados pela MSR. O 1º experimento foi um planejamento fatorial fracionado 26-2. O 2º foi baseado nas variáveis significativas do 1º experimento, resultando em um planejamento fatorial completo 33-0. Verificou-se, do primeiro para o segundo, aumento na produtividade do PHB de 12 para 45 mg g-1 de célula h-1 e do alginato de 100 para 1.600 mg g-1 de célula h-1. A produtividade de ambos os compostos foi máxima na temperatura de incubação de 62ºC, no menor tempo de incubação (18h e na concentração de sacarose de 11 g L-1. Em ambos os experimentos, o PHB extraído apresentou pureza de 94%.

  5. [Effectiveness of the biological purification of sewage for removing carcinogenic polycyclic aromatic hydrocarbons]. (United States)

    Dikun, P P; Baranova, L N; Lomova, M A


    The correlation between biological purification of sewage and concentrations of carcinogens in it is studied. The balance between the income and outcome of benza(a)pyrene (BP) with water and surplus active mud of paper in aero-tanks, performed under both industrial and laboratory conditions is examined. It is demonstrated that BP is not destroyed by biological purification but is accumulated by the active sludge cells. Thus, the biologic purification of sewage prevents from pollution by carcinogenic polycyclic hydrocarbons, but does not remove these agents from the environment.

  6. Morphological, biochemical and molecular characterization of twelve nitrogen-fixing bacteria and their response to various zinc concentration. (United States)

    Dadook, Mohammad; Mehrabian, Sedigheh; Salehi, Mitra; Irian, Saeed


    Zinc is an essential micronutrient used in the form of zinc sulfate in fertilizers in the agriculture production system. Nitrogen-fixing microorganisms are also of considerable value in promoting soil fertility. This study aimed to investigate the degree of sensitivity to varying concentrations of zinc, in the form of ZnSO4, in different strains of Azotobacter chroococcum in a laboratory environment. To isolate A. chroococcum strains, soil samples were collected from wheat, corn and asparagus rhizospheres and cultured in media lacking nitrogen at 30˚C for 48 hours. Strains were identified based on morphological and biochemical characteristics. The presence of the nitrogenase enzyme system was confirmed by testing for the presence of the nifH gene using PCR analysis. The minimum inhibitory concentration (MIC) and optimal zinc concentration for the growth of each strain was determined. A total of 12 bacterial strains were isolated from six different soil samples. A. chroococcum strains were morphologically and biochemically characterized. The presence of the nifH gene was confirmed in all the strains. MIC and the optimal zinc concentration for bacterial growth were 50 ppm and 20 ppm, respectively. It was concluded that increasing the concentration of zinc in the agricultural soil is harmful to beneficial microorganisms and reduces the soil fertility. A 20-ppm zinc concentration in soil is suggested to be optimal.

  7. Salinity tolerance of Dodonaea viscosa L. inoculated with plant growth-promoting rhizobacteria: assessed based on seed germination and seedling growth characteristics

    Directory of Open Access Journals (Sweden)

    Yousefi Sonia


    Full Text Available The study was conducted to evaluate the potential of different strains of plant growth-promoting rhizobacteria (PGPR to reduce the effects of salinity stress on the medicinal hopbush plant. The bacterium factor was applied at five levels (non-inoculated, inoculated by Pseudomonas putida, Azospirillum lipoferum + Pseudomonas putida, Azotobacter chroococcum + Pseudomonas putida, and Azospirillum lipoferum + Azotobacter chroococcum + Pseudomonas putida, and the salinity stress at six levels: 0, 5, 10, 15, 20, and 50 dS m-1. The results revealed that Pseudomonas putida showed maximal germination percentage and rate at 20 dS m-1 (18.33% and 0.35 seed per day, respectively. The strongest effect among the treatments was obtained with the treatment combining the given 3 bacteria at 15 dS m-1 salinity stress. This treatment increased the root fresh and dry weights by 31% and 87.5%, respectively (compared to the control. Our results indicate that these bacteria applied on hopbush affected positively both its germination and root growth. The plant compatibility with the three bacteria was found good, and the treatments combining Pseudomonas putida with the other one or two bacteria discussed in this study can be applied in nurseries in order to restore and extend the area of hopbush forests and akin dry stands.

  8. [Study on extraction and purification technology of Hubei ophiopogon saponins]. (United States)

    Lin, Yun-Han; Li, Chong-Ming; Li, Xiao-Dong; Xiang, Yang; Zhang, Ya-Qin; Zhang, Xiao-Cun; Liu, Xia


    To explore the extraction and purification technology of total saponins from the effective parts of Liriope spicata. Orthogonal design was used. Macroporous resin was selected to separate and purify total saponin from the effective parts of Liriope spicata. The process validation was conducted. The total saponins was determined by Ultraviolet Spectrophotometry. The optimal extraction conditions were as follows: 10 times the amount of ethanol (70%) for each occasion and hot reflux (3 x 2 h). Total saponins was purified by D101 macroporous resin. The concentration of eluting ethanol was 50% - 70%. Ethanol (70%) was selected as the best eluent. The result of process validation was consistent with the study. The process is simple and stable enough to significantly improve the extraction rate of the effective parts. The study can provide reference for the research and production of effective parts of traditional Chinese medicine such as Liriope spicata.

  9. [Separation and purification of proanthocyanidins from rose by macroporous resins]. (United States)

    Yao, Yu-cui; Li, Xiang-rong


    To study the separation and purification technology for proanthocyanidins from rose and establish the best operating conditions. Evaluated by static adsorption capacity and elution ratio, five types of macroporous resins including D101, D1300, NKA, AB-8, NKA-II were tested to separate and purify proanthocyanidins. And evaluated by product purity, the concentration of extract sample, pH of extract sample, concentration of eluant and flow rate of elution had been investigated. D101 type macroporous resin showed the best property and was suitable for purifying proanthocyanidins from rose. The best operating conditions were as follows: 1.25 mg/ml as the concentration of extract sample, 2 as the pH value of extract sample, 70% ethanol equeous solution as the eluant and 2 ml/min as the flow rate of elution. This study can supply a method to separate and purify proanthocyanidins from rose.

  10. Purification Of Water From Nsukka Water Pond Using Solar Still.

    Directory of Open Access Journals (Sweden)

    Ugwuoke E.C


    Full Text Available Abstract This work presents the analysis of a solar water distillation system. There is important need for good drinking water in the world today due to harmful effect of water borne diseases. Most water from rivers ponds seas are either salty or brackish and require purification before drinking. The water used in this work is collected from pond at Nsukka Urban and the experiment was performed at University of Nigeria Nsukka. Twenty litres of water was used for the experiment and 4 litres was obtained as the maximum volume after 10 days .The average temperature recorded during the experiment was 29C. The chemical and physical properties of the distillate correspond to world Health Organization Standard.

  11. Mechanical damage and corrosion in the primary system purification cooler

    International Nuclear Information System (INIS)

    Sainz, R.A.; Fiorini, R.H.; Semino, C.J.


    Through the routine measurements of tritium activity and isotopic content in the exchanger's cooling water, a loss of heavy water was detected. During the decommissioning programmed for October 1986, the equipment was dismantled and the tubes losses were identified through helium fugue tests; subsequently, a 100% inspection of the tubes by atmospheric currents were performed, verifying reductions exceeding 50% of the original width in 70 tubes of the first section at the top plate level. These indications were verified through the study of the two extracted tubes, one of them observing a passing failure where marks appeared at all levels of the support and corrosion plates through splits at the top plates level. The corrosion causes were due to the low cooling flow which results from the primary system's reduction regarding the purification flow design, thus permitting the deposits accumulation. (Author)

  12. CAREM 25: Suppression pool cooling and purification system

    International Nuclear Information System (INIS)

    Carlevaris, Rodolfo; Patrignani, Alberto; Vindrola, Carlos; Palmerio, Hector D.; Quiroz, Horacio; Ramilo, Lucia B.


    The suppression pool cooling and purification system has the following main functions: purify and cool water from the suppression pool, cool and send water to the residual heat extraction system, and transfer water to the fuel element transference channel. In case of Loss of Coolant Accident (LOCA), the system sends water from the suppression pool to the spray network, thus cooling and reducing pressure in the primary containment. The system has been designed in accordance with the requirements of the following standards: ANSI/ANS 52.1; ANSI/ANS 57.2; ANSI/ANS 56.2; ANSI/ANS 59.1; ANSI/ANS 58.3; ANSI/ANS 58.9; and ANSI/ANS 56.5. The design of the system fulfils all the assigned functions. (author)

  13. CAREM-25. Suppression Pool Cooling and Purification System

    International Nuclear Information System (INIS)

    Carlevaris, Rodolfo; Palmerio, D.; Patrignani, A.; Quiroz, H.; Ramilo, L.; Vindrola, C.


    The Suppression Pool Cooling and Purification System has the following main functions: purify and cool water from the Suppression Pool, cool and send water to the Residual Heat Extraction System, and transfer water to the Fuel Element Transference Channel. In case of Loss of Coolant Accident (LOCA), the system sends water from the Suppression Pool to the spray network, thus cooling and reducing pressure in the primary containment.The system has been designed in accordance with the requirements of the following standards ANSI/ANS 52.1 [1], ANSI/ANS 57.2 [2], ANSI/ANS 56.2 [3], ANSI/ANS 59.1 [4] ANSI/ANS 58.3 [5], ANSI/ANS 58.9 [6], and ANSI/ANS 56.5 [7]. The design of the system fulfils all the assigned functions

  14. Extraction and Purification of Phlorotannins from Brown Algae. (United States)

    Gall, Erwan Ar; Lelchat, Florian; Hupel, Mélanie; Jégou, Camille; Stiger-Pouvreau, Valérie


    The interest in the physiological roles and bioactivities of plant phenols has increased over the past decades. In seaweeds, many investigations have dealt with phenolic compounds of Phaeophyceae (phlorotannins), even though little is known so far about the ecophysiological variations of their pool or their biosynthetic pathways. We describe here a simple procedure based on the use of water-organic solvent mixtures for the extraction of phlorotannins. Crude extracts are semi-purified and fractionated by separating methods based on both the polarity and the molecular size of compounds. Phenols are then quantified by the Folin-Ciocalteu method and their radical-scavenging activity is characterized using the DPPH test. All along the purification process of phenolic compounds, the efficiency of separation is assessed by (1)H-NMR.

  15. Novel peptide ligand with high binding capacity for antibody purification

    DEFF Research Database (Denmark)

    Lund, L. N.; Gustavsson, P. E.; Michael, R.


    commonly used capture step in mAb down stream processing; however, the use of Protein A chromatography is less attractive due to toxic ligand leakage as well as high cost. Whether used as an alternative to the Protein A chromatographic media or as a subsequent polishing step, small synthetic peptide...... ligand for purification of human IgG. Immobilized on WorkBeads, an agarose-based base matrix from Bio-Works, the ligand has a dynamic binding capacity of up to 48 mg/mL and purifies IgG from harvest cell culture fluid with purities and recovery of >93%. The binding affinity is similar to 10(5) M-1...

  16. Purification and characterization of amine oxidase from soybean seedlings. (United States)

    Vianello, F; Di Paolo, M L; Stevanato, R; Gasparini, R; Rigo, A


    A simple and rapid procedure for purification of soybean seedling amine oxidase is reported. The crude enzyme, obtained by ammonium sulfate fractionation was purified by ion-exchange chromatography on a cellulose phosphate column and batch affinity chromatography on 6-aminohexyl-Sepharose. Cyclohexylamine, a competitive inhibitor, was utilized to elute the enzyme. A homogeneous enzyme was obtained with a yield higher than 25%, the content of minor components being lauryl sulfate-polyacrylamide gel electrophoresis. The enzyme is a dimer and contains two Cu2+ ion per molecule. Its EPR spectrum is typical of Cu2+ in a tetragonal symmetry. The enzyme oxidizes cadaverine at high rate, the specific activity being 4.3 mukat/mg. Molecular, spectroscopic, and kinetic properties of this enzyme are reported.

  17. Simplified Method for Rapid Purification of Soluble Histones

    Directory of Open Access Journals (Sweden)

    Nives Ivić


    Full Text Available Functional and structural studies of histone-chaperone complexes, nucleosome modifications, their interactions with remodelers and regulatory proteins rely on obtaining recombinant histones from bacteria. In the present study, we show that co-expression of Xenopus laevis histone pairs leads to production of soluble H2AH2B heterodimer and (H3H42 heterotetramer. The soluble histone complexes are purified by simple chromatographic techniques. Obtained H2AH2B dimer and H3H4 tetramer are proficient in histone chaperone binding and histone octamer and nucleosome formation. Our optimized protocol enables rapid purification of multiple soluble histone variants with a remarkable high yield and simplifies histone octamer preparation. We expect that this simple approach will contribute to the histone chaperone and chromatin research. This work is licensed under a Creative Commons Attribution 4.0 International License.

  18. A purification process for an inert gas system

    International Nuclear Information System (INIS)

    Raj, S.S.; Samanta, S.K.; Jain, N.G.; Deshingkar, D.S.; Ramaswamy, M.


    Special inert atmosphere is desired inside hot cells used for handling radioactive materials. In this report, details of experiments conducted to generate data required for the design of a system for maintaining very low levels of organic and acid vapours, oxygen and moisture in a nitrogen gas inert atmosphere, are described. Several grades of activated charcoals impregnated with 1% KOH were studied for the adsorption of acidic and organic vapours. A Pd/Al 2 O 3 catalyst was developed to remove oxygen with greater than 90% efficiency. For the removal of moisture, a regenerable molecular sieve 4A dual-bed was provided. Based on the performance data thus generated, an integrated purification system for nitrogen gas is proposed. (author)

  19. Design and performance tests of OGL-1 purification system

    International Nuclear Information System (INIS)

    Tanaka, Toshiyuki; Takahashi, Hidetake; Tone, Hirohito; Matsunaga, Sakuro


    The in-pile helium gas loop, OGL-1, was installed in JMTR at Oarai site of JAERI in 1976. Two types of the test fuel have been irradiated successfully. Low levels of both radioactivity and chemical contaminants show the integrity of VHTR test fuels and also no deterioration of graphite and other structural materials of high temperature components. The purification system consisting of a series of adsorbent beds works well to maintain the concentration of impurities in the coolant in the ranges of less than 10 vpm and to remove radioactive fission products from the main loop (10 -5 mu ci/ml at loop conditions). Plateout activities are low enough to assure the capability of access to the system for maintenance after reactor shutdown. (author)

  20. Surfactin – A Review on Biosynthesis, Fermentation, Purification and Applications

    Directory of Open Access Journals (Sweden)

    Nikhil S. Shaligram


    Full Text Available Surfactin, a bacterial cyclic lipopeptide, is produced by various strains of Bacillus subtilis and is primarily recognized as one of the most effective biosurfactants. It has the ability to reduce surface tension of water from 72 to 27 mN/m at a concentration as low as 0.005 %. The structure of surfactin consists of seven amino acids bonded to the carboxyl and hydroxyl groups of a 14-carbon fatty acid. Surfactin possesses a number of biological activities such as the ability to lyse erythrocytes, inhibit clot formation, lyse bacterial spheroplasts and protoplasts, and inhibit cyclic 3',5-monophosphate diesterase. The high cost of production and low yields have limited its use in various commercial applications. Both submerged and solid-state fermentation have been investigated with the mutational approach to improve the productivity. In this review, current state of knowledge on biosynthesis of surfactin, its fermentative production, purification, analytical methods and biomedical applications is presented.

  1. Partial purification and characterization of the glucagon receptor

    International Nuclear Information System (INIS)

    Horuk, R.; Wright, D.E.


    Specific labeling of liver plasma membrane glucagon receptors has been achieved by the photoincorporation of a 125 I-labeled photoderivative of glucagon, Nsup(E)-4-azidophenylamidinoglucagon. Identification of glucagon receptors was facilitated by irradiating membranes in the presence of excess unlabeled glucagon. Isoelectric focusing of radioiodinated membrane proteins revealed one major band of glucagon displaceable material which had an isoelectric point of 5.85. When this material was isolated and run on SDS-polyacrylamide gels a major labeled band of Msub(r) 55000 was obtained which had properties consistent with those of the glucagon receptor. These studies indicate that a purification of the glucagon receptor of >700-fold can be attained through the use of isoelectric focusing and SDS-polyacrylamide electrophoresis. (Auth.)

  2. Purification and deodorization of structured lipids by short path distillation

    DEFF Research Database (Denmark)

    Xu, Xuebing; Jacobsen, Charlotte; Nielsen, Nina Skall


    Purification of structured lipids (SL), produced from lipase- catalyzed acidolysis of rapeseed oil and capric acid, and deodorization of randomized SL, produced from chemical randomization of fish oil and tricaprin, were studied in a bench-scale short path distillation (SPD). SL obtained from....... An applicable parameter zone was created to obtain a certain FFA (0.5% for example) content. In general, conditions that result in a lower FFA content will lead to a higher loss of tocopherols. In most parts of the parameter zone, 50% loss of tocopherols will be expected. The deodorization study of randomized...... SL from fish oils and tricaprin indicated that SPD in comparison with batch deodorization gave a product of a poorer sensoric quality...

  3. Pressure regulation in the dry-boxes. Argon purification

    International Nuclear Information System (INIS)

    Pascard, R.; Fabre, R.


    Each dry-box is equipped with an autonomous installation for circulation and purification of argon and for pressure regulation. This installation consists essentially of a ballast tank, a compressor and two valves electromagnetically controlled by a contact manometer. The compressor and the valves are enclosed in the tank to form a system as compact as possible. The argon is purified by passing it over a furnace filled with titanium-zirconium turnings brought to about 800 deg. C, branching off the main system. With this set-up as well as the automatic maintenance of a constant depression in the box, a quality of argon is obtained whose oxygen contact is undetectable by the manganous hydroxide method. (author) [fr

  4. Membranes with Surface-Enhanced Antifouling Properties for Water Purification (United States)

    Shahkaramipour, Nima; Tran, Thien N.; Ramanan, Sankara; Lin, Haiqing


    Membrane technology has emerged as an attractive approach for water purification, while mitigation of fouling is key to lower membrane operating costs. This article reviews various materials with antifouling properties that can be coated or grafted onto the membrane surface to improve the antifouling properties of the membranes and thus, retain high water permeance. These materials can be separated into three categories, hydrophilic materials, such as poly(ethylene glycol), polydopamine and zwitterions, hydrophobic materials, such as fluoropolymers, and amphiphilic materials. The states of water in these materials and the mechanisms for the antifouling properties are discussed. The corresponding approaches to coat or graft these materials on the membrane surface are reviewed, and the materials with promising performance are highlighted. PMID:28273869

  5. Continuous chemical cold traps for reprocessing off-gas purification

    International Nuclear Information System (INIS)

    Henrich, E.; Bauder, U.; Steinhardt, H.J.; Bumiller, W.


    Absorption of nitrogen oxides and iodine from simulated reprocessing plant off-gas streams has been studied using nitric acid and nitric acid/hydrogen peroxide mixtures at low temperatures. The experiments were carried out at the laboratory and on the engineering scale. The pilot plant scale column has 0.8 m diameter and 16 absorption plates at 0.2 m spacing. Cooling coils on the plates allow operating temperatures down to -60 0 C. The NO concentration in the feed gas usually has been 1% by volume and the flow rate 4-32 m 3 (STP) per hour. The iodine behavior has been studied using I-123 tracer. Results of the study are presented. The chemistry of the processes and the advantages and disadvantages in correlation to the various applications for an off-gas purification in a reprocessing plant are compared and discussed. The processes are compatible with the PUREX process and do not produce additional waste

  6. Purification of free arginine from chickpea (Cicer arietinum) seeds. (United States)

    Cortés-Giraldo, Isabel; Megías, Cristina; Alaiz, Manuel; Girón-Calle, Julio; Vioque, Javier


    Chickpea is a grain legume widely consumed in the Mediterranean region and other parts of the world. Chickpea seeds are rich in proteins but they also contain a substantial amount of free amino acids, especially arginine. Hence chickpea may represent a useful source of free amino acids for nutritional or pharmaceutical purposes. Arginine is receiving great attention in recent years because it is the substrate for the synthesis of nitric oxide, an important signaling molecule involved in numerous physiological and pathological processes in mammals. In this work we describe a simple procedure for the purification of arginine from chickpea seeds, using nanofiltration technology and an ion-exchange resin, Amberlite IR-120. Arginine was finally purified by precipitation or crystallization, yielding preparations with purities of 91% and 100%, respectively. Chickpea may represent an affordable green source of arginine, and a useful alternative to production by fermentation or protein hydrolysis. Copyright © 2015 Elsevier Ltd. All rights reserved.


    Energy Technology Data Exchange (ETDEWEB)

    Korinko, P; T. Adams


    Development of advanced hydrogen separation membranes in support of hydrogen production processes such as coal gasification and as front end gas purifiers for fuel cell based system is paramount to the successful implementation of a national hydrogen economy. Current generation metallic hydrogen separation membranes are based on Pd-alloys. Although the technology has proven successful, at issue is the high cost of palladium. Evaluation of non-noble metal based dense metallic separation membranes is currently receiving national and international attention. The focal point of the reported work was to evaluate two different classes of materials for potential replacement of conventional Pd-alloy purification/diffuser membranes. Crystalline V-Ni-Ti and Amorphous Fe- and Co-based metallic glass alloys have been evaluated using gaseous hydrogen permeation testing techniques.

  8. Analytical methods used in plutonium purification cycles by trilaurylamine

    International Nuclear Information System (INIS)

    Perez, J.J.


    The utilisation of trilaurylamine as a solvent extractant for the purification of plutonium has entailed to perfect a set of analytical methods which involves, various techniques. The organic impurities of the solvent can be titrated by gas-liquid chromatography. The titration of the main degradation product, the di-laurylamine, can be accomplished also by spectro-colorimetry. Potentiometry is used for the analysis of the different salts of amine-nitrate-sulfate-bisulfate as also the extracted nitric acid. The determination of the nitrate in aqueous phase is carried out by constant current potentiometry. The range of application, the accuracy and the procedure of these analysis are related in the present report. (author) [fr

  9. Influence of Water Salinity on Air Purification from Hydrogen Sulfide

    Directory of Open Access Journals (Sweden)

    Leybovych L.I.


    Full Text Available Mathematical modeling of «sliding» water drop motion in the air flow was performed in software package FlowVision. The result of mathematical modeling of water motion in a droplet with diameter 100 microns at the «sliding» velocity of 15 m/s is shown. It is established that hydrogen sulfide oxidation occurs at the surface of phases contact. The schematic diagram of the experimental setup for studying air purification from hydrogen sulfide is shown. The results of the experimental research of hydrogen sulfide oxidation by tap and distilled water are presented. The dependence determining the share of hydrogen sulfide oxidized at the surface of phases contact from the dimensionless initial concentration of hydrogen sulfide in the air has been obtained.

  10. Antibodies and genetically engineered related molecules: production and purification. (United States)

    Roque, A Cecília A; Lowe, Christopher R; Taipa, M Angela


    Antibodies and antibody derivatives constitute 20 % of biopharmaceutical products currently in development, and despite early failures of murine products, chimeric and humanized monoclonal antibodies are now viable therapeutics. A number of genetically engineered antibody constructions have emerged, including molecular hybrids or chimeras that can deliver a powerful toxin to a target such as a tumor cell. However, the general use in clinical practice of antibody therapeutics is dependent not only on the availability of products with required efficacy but also on the costs of therapy. As a rule, a significant percentage (50-80%) of the total manufacturing cost of a therapeutic antibody is incurred during downstream processing. The critical challenges posed by the production of novel antibody therapeutics include improving process economics and efficiency, to reduce costs, and fulfilling increasingly demanding quality criteria for Food and Drug Administration (FDA) approval. It is anticipated that novel affinity-based separations will emerge from the development of synthetic ligands tailored to specific biotechnological needs. These synthetic affinity ligands include peptides obtained by synthesis and screening of peptide combinatorial libraries and artificial non-peptidic ligands generated by a de novo process design and synthesis. The exceptional stability, improved selectivity, and low cost of these ligands can lead to more efficient, less expensive, and safer procedures for antibody purification at manufacturing scales. This review aims to highlight the current trends in the design and construction of genetically engineered antibodies and related molecules, the recombinant systems used for their production, and the development of novel affinity-based strategies for antibody recovery and purification.

  11. Nicotinamide riboside phosphorylase from beef liver: purification and characterization. (United States)

    Imai, T; Anderson, B M


    Nicotinamide riboside phosphorylase (NR phosphorylase) from beef liver has been purified to apparent homogeneity at 300-fold purification with a 35% yield. Kinetic constants for the enzyme-catalyzed phosphorolysis were as follows Knicotinamide riboside, 2.5 +/- 0.4 mM; Kinorganic phosphate, 0.50 +/- 0.12 mM; Vmax, 410 +/- 30 X 10(-6) mol min-1 mg protein-1, respectively. The molecular weights of the native enzyme and subunit structure were determined to be 131,000 and 32,000, respectively, suggesting the beef liver NR phosphorylase to be tetrameric in structure and consistent with the presence of identical subunits. The amino acid composition was shown to be very similar to that reported for human erythrocyte purine-nucleoside phosphorylase but differing considerably from that found for rat liver purine-nucleoside phosphorylase. In addition to catalytic activity with nicotinamide riboside, the beef liver enzyme catalyzed a phosphorolytic reaction with inosine and guanosine exhibiting activity ratios, nicotinamide riboside:inosine: guanosine of 1.00:0.35:0.29, respectively. These ratios of activity remained constant throughout purification of the beef liver enzyme and no separation of these activities was detected. Phosphorolysis of nicotinamide riboside was inhibited competitively by inosine (Ki = 75 microM) and guanosine (Ki = 75 microM). Identical rates of thermal denaturation of the beef liver enzyme were observed when determined for the phosphorolysis of either nicotinamide riboside or inosine. These observations coupled with studies of pH and specific buffer effects indicate the phosphorolysis of nicotinamide riboside, inosine, and guanosine to be catalyzed by the same enzyme.

  12. Direct osmosis method of purification and desalination of drinking water

    International Nuclear Information System (INIS)

    Khaydarov, R.A.; Khaydarov, R.R.


    Full text: Drinking water quality is one of the general factors influencing people's health. The human activity in industry and agriculture has led to pollution of the environment: soil, air, both surface and ground waters that are polluted with chemical substances. It has a disastrous effect on the health of the population, especially of children. At present, the known equipment, based on ion exchange, electrodialysis and reverse osmosis, require great expense, energy expenditures, and highly qualified personnel that are inaccessible to the population especially living in remote regions. Methods, which are usually used in water supplying plants, cannot remove spore forms of bacteria and many types of chemical substances. The purpose of this Project is to create an absolutely new method for purification of drinking water from chemical and biological agents. The method is based on using direct osmosis process that removes all contaminants except one and removing last contaminant. This method will be used for making new low energy-consuming and cheap mini-systems for individual and collective use for desalination of drinking water and purification from bacteria, radionuclides, heavy metal ions, and organic contaminants. Preliminary experiments and calculations conducted in Uzbekistan show that the energy consumption is 0.8 MW per 1 m 3 of water. Advantage of the method is low energy consumption, potentially purifying water without pretreatment and removing different types of bacteria including spore forms, radionuclides, heavy metal ions, organic contaminants. Devices can be powered by solar units in remote locations. The purpose of this work is further elaboration of this technology creation of new method and its accommodation to conditions of different countries. Test models will be made and tested in laboratories of interested countries

  13. Research of Biogas Purification Using Microalgae Monoraphidium Griffithii Suspension

    Directory of Open Access Journals (Sweden)

    Živilė Bingelytė


    Full Text Available Using biogas instead of fossil fuels decreases pollutants such as solid particles, sulphur dioxide, nitrogen oxides concentrations in the environment. Green energy and development of relevant infrastructure improves air quality considerably. Chemical, physical, biological methods are used for biogas purification. The main difficulties using biological methods are selection of suitable microorganisms’ suspensions and making optimal conditions in photobioreactor. Different origin and structure microalgae suspensions are used applying biological treatment methods. Monoraphidium griffithi, which is widespread in fresh water, has relatively high potential. Microalgae’ cultures absorb the main components of biogas – carbon dioxide (CO2 and hydrogen sulphide (H2S. Absorbtion processes are based on photosynthesis. Microalgae absorb specific components of biogas when there are suitable light source and nutrient solvent. The main purposes of the research are to asses emission of biogas using different substrates (chicken manure and wastewater sludge. Also, it was measured main physical and chemical characteristics of both substrates: acidicy, temperature, redox potential, conductivity, biohemical oxygen demand. According results of the research, emission from wastewater sludge is greater than from chicken manure so sludge was chosen in teh next stage of the research. The next stage – asssessment of purification efficienty using Monoraphidium grifftihii suspension. Raw biogas was supplied to photobioreacor (with microalgae suspension. Alterations of methane, carbon dioxide, oxygen, hydrogen sulphide concentrations were measured precisely. According to results concentration of methane in the beginning of the researc was 62%, after 35 days – 69%. Meanwhile carbon dioxide – 37% and 31% by analogy. Experimental research alows to assess Monoraphidium griffithi absorption capacity of ballast components. Results were compared to different scientists

  14. Preliminary studies on the purification of a monoclonal antibody by affinity precipitation with Eudragit S-100. (United States)

    Taipa, M A; Kaul, R; Mattiasson, B; Cabral, J M


    A simple procedure for the purification of an IgG-type monoclonal antibody by affinity precipitation using Eudragit S-100 is presented. The ligand, a microbial lipase previously used as antigen, was coupled to the polymer at a concentration of 40 mg lipase/g Eudragit. This macroligand was reversibly precipitated by manipulating the pH at values higher and lower than 4.8. The effects of polymer concentration and dilution of hybridoma culture supernatant on the overall precipitation process were evaluated. The best purification factor was achieved with a polymer concentration of 0.1% (w/v) and a supernatant dilution of 1:3. The preliminary studies reported here enabled the purification of a monoclonal antibody in one step with an activity yield (by ELISA) of 50%-55% and a purification factor of ca 6.

  15. Comparative analysis of environmental DNA extraction and purification methods from different humic acid rich soils

    CSIR Research Space (South Africa)

    Lakay, FM


    Full Text Available Three different soil DNA isolation and four purification strategies were compared on different soil samples with variable rates of success. Bead beating extraction gave significantly higher DNA yields than microwave-based and liquid nitrogen...

  16. Experimental research on photocatalytic oxidation air purification technology applied to aircraft cabins

    DEFF Research Database (Denmark)

    Sun, Yuexia; Fang, Lei; Wyon, David Peter


    The experiment presented in this report was performed in a simulated aircraft cabin to evaluate the air cleaning effects of two air purification devices that used photocatalytic oxidation (PCO) technology. Objective physical, chemical and physiological measurements and subjective human assessments...

  17. Sodium tetrathionate effect on papain purification from different Carica papaya latex crude extracts. (United States)

    Llerena-Suster, Carlos R; Priolo, Nora S; Morcelle, Susana R


    Papain from latex of Carica papaya was purified up to matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry homogeneity by salt precipitation from two different crude extract sources: a refined preparation obtained in our laboratory and a commercial one. Sodium tetrathionate was tested in the purification process to preserve the enzymatic activity of the peptidase. Purification was checked by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and cation exchange chromatography, using commercial pure papain as standard for a rapid comparison. The best purification yields (3.4%) were obtained in presence of 30 mM sodium tetrathionate for the crude extract prepared in our laboratory. The described purification method proved to be robust and reliable to obtain pure papain on a preparative scale.

  18. Tensile deformation of polytetrafluoroethylene hollow fiber membranes used for water purification. (United States)

    Yonezu, Akio; Iio, Shouichi; Itonaga, Takehiro; Yamamura, Hiroshi; Chen, Xi


    The tensile deformation behavior of polytetrafluoroethylene (PTFE) hollow fiber membranes is studied. PTFE membranes at present have sub-micron pores with an open cell structure, which plays a critical role in water purification. One of the main challenges in water purification is that the pore structure becomes covered with biofouling, leading to blocked pores. To maintain the capacity for water purification, physical cleaning along with mechanical deformation is usually conducted. Thus, it is crucial to understand the mechanical properties, in particular the deformation behavior, of the membrane fibers. Using uniaxial tension experiments, we established a fundamental discrete model to describe the deformation behavior of a porous structure using a finite element method. The present model enables the prediction of the macroscopic deformation behavior of the membrane, by taking into account the changes of pore structure. The insight may be useful for porous membrane fabrication and provide insights for the reliable operation of water purification.

  19. Virus purification by CsCl density gradient using general centrifugation. (United States)

    Nasukawa, Tadahiro; Uchiyama, Jumpei; Taharaguchi, Satoshi; Ota, Sumire; Ujihara, Takako; Matsuzaki, Shigenobu; Murakami, Hironobu; Mizukami, Keijirou; Sakaguchi, Masahiro


    Virus purification by cesium chloride (CsCl) density gradient, which generally requires an expensive ultracentrifuge, is an essential technique in virology. Here, we optimized virus purification by CsCl density gradient using general centrifugation (40,000 × g, 2 h, 4 °C), which showed almost the same purification ability as conventional CsCl density gradient ultracentrifugation (100,000 × g, 1 h, 4 °C) using phages S13' and φEF24C. Moreover, adenovirus strain JM1/1 was also successfully purified by this method. We suggest that general centrifugation can become a less costly alternative to ultracentrifugation for virus purification by CsCl densiy gradient and will thus encourage research in virology.

  20. Undulative induction electron accelerator for the waste and natural water purification systems

    CERN Document Server

    Kulish, Victor V; Gubanov, I V


    The project analysis of Undulative Induction Accelerator (EH - accelerator) for the waste and natural water purification systems is accomplished. It is shown that the use of the four-channel design of induction block and the standard set of auxiliary equipment (developed earlier for the Linear Induction Accelerators - LINACs) allow to construct commercially promising purification systems. A quality analysis of the accelerator is done and the optimal parameters are chosen taking into account the specific sphere of its usage.

  1. Relationship Between Growth of AIgae and Water Purification in a Slow Sand Filter in Summe


    中本, 信忠; 池田, 大介; 田口, 香代; 山本, 満寿夫; 松田, 卓也


    The effects of water depth on the growth of algae and on the purification capacity of water in slow sand filters in summer were studied. Filamentous algae grew well in a shallow filter pond. The higher removal rates of available nutrients and dissolved organic carbon in a raw water were observed in the filteration of a shallow filter pond. Importance of algae as a nutrient assimilator and as an oxygen producer in the purification process was discussed.

  2. An improved strategy for tandem affinity purification-tagging of Schizosaccharomyces pombe genes


    Cipak, Lubos; Spirek, Mario; Novatchkova, Maria; Chen, Zhiming; Rumpf, Cornelia; Lugmayr, Wolfgang; Mechtler, Karl; Ammerer, Gustav; Csaszar, Edina; Gregan, Juraj


    Tandem affinity purification (TAP) is a method that allows rapid purification of native protein complexes. We developed an improved technique to fuse the fission yeast genes with a TAP tag. Our technique is based on tagging constructs that contain regions homologous to the target gene cloned into vectors carrying a TAP tag. We used this technique to design strategies for TAP-tagging of predicted Schizosaccharomyces pombe genes ( To validate the approach...

  3. Small scale purification of human pituitary lutropin (hLH) for use in radioligand assays

    International Nuclear Information System (INIS)

    Schwarz, I.; Morgante, L.; Bartolini, P.


    Human lutropin (hLH) is a relatively unstable protein, which even in lyophilised form tends to dissociate into subunits during long storage periods. Considering the limited disposibilty of human pituitaries, a small-scale extraction method is proposed for radioassays. Starting from 10 and 20 hypophyses after Sephadex G 100 purification, 10 μg/gland with approximate 10% purity was obtained. After the last purification, hLH recovery was of 1.5 μg/gland. (author) [pt

  4. Experience with sodium purification on large-scale operational test loops in France

    International Nuclear Information System (INIS)

    Benoist, E.; Cambillard, M.; Quinet, J.L.


    The major characteristics of four large test loops operational in France are discussed and the sodium purification equipment used in conjunction with these test facilities and the purification monitoring devices and operational procedures are reviewed. Operating experience is also described with particular attention to three types of impurities: calcium from ''industrial grade'' sodium, oil from the mechanical pumps and hydrogen from the steam generators [fr

  5. Purification of indium, gallium, and antimony by melting with synthetic slug

    International Nuclear Information System (INIS)

    Walis, L.; Rowinska, L.; Nowicki, A.


    The tracer technique has been used for metal purification process control. The indium, gallium and antimony have been purified up to semiconductor purity in the melting with synthetic slug process. The 115m Cd, 59 Fe, 204 Tl. 76 As, 210 Bi have been used as tracers. The high effectiveness of proposed purification method have been found in the cases of indium and gallium. 6 refs, 2 figs, 2 tabs

  6. Partial Purification of Antimicrobial Compounds Isolated from Mycelia of Tropical Lentinus cladopus LC4




    Lentinus cladopus LC4 produced at least eight antimicrobial compounds (ACs) which are active against plant and human pathogens. Three ACs in its crude mycelial were extracted with methanol and partial purification was carried out with silicic acid column chromatography and by thin layer chromatography (PTLC). The antimicrobial activity was tested by paper disc method and antibiographic method. The chromatography purification eluted with dichloromethane containing 5% methanol gave one active f...

  7. Rapid purification of radiodinated glucagon with Sep-PakR reversed phase cartridges

    International Nuclear Information System (INIS)

    Borghi, V.C.; Nascimento, M.; Wajchenberg, B.L.


    A simple, rapid method is described for the purification of radioiodinated glucagon for use in radioimmunoassay. Samples of the radioiodinated hormone, before and after purification are analysed on polyacrylamide gel electrophoresis. The 125 I incorporation in these samples is also checked through thrichloroacetic acid precipitation. The usefulness of this labeled glucagon in radioimmunoassay is demonstrated by its biding to specific antibodies. (M.A.C.) [pt

  8. A chimeric affinity tag for efficient expression and chromatographic purification of heterologous proteins from plants

    Directory of Open Access Journals (Sweden)

    Frank eSainsbury


    Full Text Available The use of plants as expression hosts for recombinant proteins is an increasingly attractive option for the production of complex and challenging biopharmaceuticals. Tools are needed at present to marry recent developments in high-yielding gene vectors for heterologous expression with routine protein purification techniques. In this study we designed the Cysta-tag, a new purification tag for immobilized metal affinity chromatography (IMAC of plant-made proteins based on the protein-stabilizing fusion partner SlCYS8. We show that the Cysta-tag may be used to rapidly purify proteins under native conditions, and then be removed enzymatically to isolate the protein of interest. We also show that commonly used protease recognition sites for linking purification tags are differentially stable in leaves of the commonly used expression host Nicotiana benthamiana, with those linkers susceptible to cysteine proteases being less stable then serine protease-cleavable linkers. As an example we describe a Cysta-tag experimental scheme for the one-step purification of a clinically useful protein, human α1-antitrypsin, transiently expressed in N. benthamiana. With potential applicability to the variety of chromatography formats commercially available for IMAC-based protein purification, the Cysta-tag provides a convenient means for the efficient and cost-effective purification of recombinant proteins from plant tissues.

  9. Automated small‐scale protein purification and analysis for accelerated development of protein therapeutics (United States)

    LeSaout, Xavier; Costioli, Matteo; Jordan, Lynn; Lambert, Jeremy; Beighley, Ross; Provencher, Laurel; McGuire, Kevin; Verlinden, Nico; Barry, Andrew


    Small‐scale protein purification presents opportunities for accelerated process development of biotherapeutic molecules. Miniaturization of purification conditions reduces time and allows for parallel processing of samples, thus offering increased statistical significance and greater breadth of variables. The ability of the miniaturized platform to be predictive of larger scale purification schemes is of critical importance. The PerkinElmer JANUS BioTx Pro and Pro‐Plus workstations were developed as intuitive, flexible, and automated devices capable of performing parallel small‐scale analytical protein purification. Preprogrammed methods automate a variety of commercially available ion exchange and affinity chromatography solutions, including miniaturized chromatography columns, resin‐packed pipette tips, and resin‐filled microtiter vacuum filtration plates. Here, we present a comparison of microscale chromatography versus standard fast protein LC (FPLC) methods for process optimization. In this study, we evaluated the capabilities of the JANUS BioTx Pro‐Plus robotic platform for miniaturized chromatographic purification of proteins with the GE ӒKTA Express system. We were able to demonstrate predictive analysis similar to that of larger scale purification platforms, while offering advantages in speed and number of samples processed. This approach is predictive of scale‐up conditions, resulting in shorter biotherapeutic development cycles and less consumed material than traditional FPLC methods, thus reducing time‐to‐market from discovery to manufacturing. PMID:27774045

  10. Plasmid vectors for proteomic analyses in Giardia: purification of virulence factors and analysis of the proteasome. (United States)

    Jerlström-Hultqvist, Jon; Stadelmann, Britta; Birkestedt, Sandra; Hellman, Ulf; Svärd, Staffan G


    In recent years, proteomics has come of age with the development of efficient tools for purification, identification, and characterization of gene products predicted by genome projects. The intestinal protozoan Giardia intestinalis can be transfected, but there is only a limited set of vectors available, and most of them are not user friendly. This work delineates the construction of a suite of cassette-based expression vectors for use in Giardia. Expression is provided by the strong constitutive ornithine carbamoyltransferase (OCT) promoter, and tagging is possible in both N- and C-terminal configurations. Taken together, the vectors are capable of providing protein localization and production of recombinant proteins, followed by efficient purification by a novel affinity tag combination, streptavidin binding peptide-glutathione S-transferase (SBP-GST). The option of removing the tags from purified proteins was provided by the inclusion of a PreScission protease site. The efficiency and feasibility of producing and purifying endogenous recombinant Giardia proteins with the developed vectors was demonstrated by the purification of active recombinant arginine deiminase (ADI) and OCT from stably transfected trophozoites. Moreover, we describe the tagging, purification by StrepTactin affinity chromatography, and compositional analysis by mass spectrometry of the G. intestinalis 26S proteasome by employing the Strep II-FLAG-tandem affinity purification (SF-TAP) tag. This is the first report of efficient production and purification of recombinant proteins in and from Giardia, which will allow the study of specific parasite proteins and protein complexes.

  11. The effect of water purification systems on fluoride content of drinking water

    Directory of Open Access Journals (Sweden)

    Prabhakar A


    Full Text Available Objective: The purpose of the present study was to determine the effect of different water purification systems on the fluoride content of drinking water and to compare the efficacy of these water purification systems in reducing the fluoride content. Materials and Methods: Five different water purification systems were tested in this study. They were reverse osmosis, distillation, activated carbon, Reviva ® , and candle filter. The water samples in the study were of two types, viz, borewell water and tap water, these being commonly used by the people of Davangere City, Karnataka. The samples were collected before and after purification, and fluoride analysis was done using fluoride ion-specific electrode. Results: The results showed that the systems based on reverse osmosis, viz, reverse osmosis system and Reviva ® showed maximum reduction in fluoride levels, the former proving to be more effective than the latter; followed by distillation and the activated carbon system, with the least reduction being brought about by candle filter. The amount of fluoride removed by the purification system varied between the system and from one source of water to the other. Interpretation and Conclusion: Considering the beneficial effects of fluoride on caries prevention; when drinking water is subjected to water purification systems that reduce fluoride significantly below the optimal level, fluoride supplementation may be necessary. The efficacy of systems based on reverse osmosis in reducing the fluoride content of water indicates their potential for use as defluoridation devices.

  12. Continuous Purification of Colloidal Quantum Dots in Large-Scale Using Porous Electrodes in Flow Channel (United States)

    Lim, Hosub; Woo, Ju Young; Lee, Doh Chang; Lee, Jinkee; Jeong, Sohee; Kim, Duckjong


    Colloidal Quantum dots (QDs) afford huge potential in numerous applications owing to their excellent optical and electronic properties. After the synthesis of QDs, separating QDs from unreacted impurities in large scale is one of the biggest issues to achieve scalable and high performance optoelectronic applications. Thus far, however, continuous purification method, which is essential for mass production, has rarely been reported. In this study, we developed a new continuous purification process that is suitable to the mass production of high-quality QDs. As-synthesized QDs are driven by electrophoresis in a flow channel and captured by porous electrodes and finally separated from the unreacted impurities. Nuclear magnetic resonance and ultraviolet/visible/near-infrared absorption spectroscopic data clearly showed that the impurities were efficiently removed from QDs with the purification yield, defined as the ratio of the mass of purified QDs to that of QDs in the crude solution, up to 87%. Also, we could successfully predict the purification yield depending on purification conditions with a simple theoretical model. The proposed large-scale purification process could be an important cornerstone for the mass production and industrial use of high-quality QDs.

  13. Engineering a recyclable elastin-like polypeptide capturing scaffold for non-chromatographic protein purification. (United States)

    Liu, Fang; Chen, Wilfred


    Previously, we reported a non-chromatographic protein purification method exploiting the highly specific interaction between the dockerin and cohesin domains from Clostridium thermocellum and the reversible aggregation property of elastin-like polypeptide (ELP) to provide fast and cost-effective protein purification. However, the bound dockerin-intein tag cannot be completely dissociated from the ELP-cohesin capturing scaffold due to the high binding affinity, resulting in a single-use approach. In order to further reduce the purification cost by recycling the ELP capturing scaffold, a truncated dockerin domain with the calcium-coordinating function partially impaired was employed. We demonstrated that the truncated dockerin domain was sufficient to function as an effective affinity tag, and the target protein was purified directly from cell extracts in a single binding step followed by intein cleavage. The efficient EDTA-mediated dissociation of the bound dockerin-intein tag from the ELP-cohesin capturing scaffold was realized, and the regenerated ELP capturing scaffold was reused in another purification cycle without any decrease in the purification efficiency. This recyclable non-chromatographic based affinity method provides an attractive approach for efficient and cost-effective protein purification. © 2013 American Institute of Chemical Engineers.

  14. A versatile multidimensional protein purification system with full internet remote control based on a standard HPLC system. (United States)

    Riek, Uwe; Ramirez, Sacnicte; Wallimann, Theo; Schlattner, Uwe


    The standard Akta Explorer high-performance liquid chromatography (HPLC) system has limitations for the automation of multidimensional protein purification. Here, we describe simple modifications that allow for automated multidimensional purification protocols to extend the possibilities of the Akta three-dimensional purification kit in terms of column number, flexibility of volumes stocked for re-injection of samples, and available choice of buffers. These modifications do not preclude the use of standard one-dimensional purification protocols. Additionally, we demonstrate a technology for encrypted full remote control of the machine over the Internet by cost-effective use of standard asymmetric digital subscriber line (ADSL) that enables direct remote interaction with the machine without preventing local control. A 4-column purification scheme, including equilibration and cleaning in place (CIP) procedures, was implemented on such a system. It significantly increased reproducibility and shortened processing time by 85%, as compared with manual operation, thus allowing for automated protein purification overnight.

  15. Over-Expression, Purification and Crystallization of Human Dihydrolipoamide Dehydrogenase (United States)

    Hong, Y. S.; Ciszak, Ewa; Patel, Mulchand


    Dehydrolipoamide dehydrogenase (E3; dihydrolipoan-tide:NAD+ oxidoreductase, EC is a common catalytic component found in pyruvate dehydrogenase complex, alpha-ketoglutarate dehydrogenase complex, and branched-chain cc-keto acid dehydrogenase complex. E3 is also a component (referred to as L protein) of the glycine cleavage system in bacterial metabolism (2). Active E3 forms a homodimer with four distinctive subdomain structures (FAD binding, NAD+ binding, central and interface domains) with non-covalently but tightly bound FAD in the holoenzyme. Deduced amino acids from cloned full-length human E3 gene showed a total of 509 amino acids with a leader sequence (N-terminal 35 amino acids) that is excised (mature form) during transportation of expressed E3 into mitochondria membrane. So far, three-dimensional structure of human E3 has not been reported. Our effort to achieve the elucidation of the X-ray crystal structure of human E3 will be presented. Recombinant pPROEX-1 expression vector (from GIBCO BRL Life Technologies) having the human E3 gene without leader sequence was constructed by Polymerase Chain Reaction (PCR) and subsequent ligation, and cloned in E.coli XL1-Blue by transformation. Since pPROEX-1 vector has an internal His-tag (six histidine peptide) located at the upstream region of a multicloning site, one-step affinity purification of E3 using nickelnitriloacetic acid (Ni-NTA) agarose resin, which has a strong affinity to His-tag, was feasible. Also a seven-amino-acid spacer peptide and a recombinant tobacco etch virus protease recognition site (seven amino acids peptide) found between His-tag and first amino acid of expressed E3 facilitated the cleavage of His-tag from E3 after the affinity purification. By IPTG induction, ca. 15 mg of human E3 (mature form) was obtained from 1L LB culture with overnight incubation at 25C. Over 98% of purity of E3 from one-step Ni-NTA agarose affinity purification was confirmed by SDS-PAGE analysis. For

  16. Expression, Purification, Crystallization and Preliminary X-ray Analysis of Pseudomonas fluorescens AlgK

    Energy Technology Data Exchange (ETDEWEB)

    Keiski,C.; Yip, P.; Robinson, H.; Burrows, L.; Howell, P.


    AlgK is an outer-membrane lipoprotein involved in the biosynthesis of alginate in Pseudomonads and Azotobacter vinelandii. A recombinant form of Pseudomonas fluorescens AlgK with a C-terminal polyhistidine affinity tag has been expressed and purified from the periplasm of Escherichia coli cells and diffraction-quality crystals of AlgK have been grown using the hanging-drop vapour-diffusion method. The crystals grow as flat plates with unit-cell parameters a = 79.09, b = 107.85, c = 119.15 {angstrom}, = 96.97{sup o}. The crystals exhibit the symmetry of space group P2{sub 1} and diffract to a minimum d-spacing of 2.5 {angstrom} at Station X29 of the National Synchrotron Light Source, Brookhaven National Laboratory. On the basis of the Matthews coefficient (V{sub M} = 2.53 {angstrom}{sup 3} Da{sup -1}), four protein molecules are estimated to be present in the asymmetric unit.

  17. Pyridine nucleotide transhydrogenase

    NARCIS (Netherlands)

    Broek, van den H.W.J.


    A method for the isolation and purification of a reversible transhydrogenase from Azotobacter vinelandii is described (CHAPTER 3). The purification of the enzyme is hampered by association-dissociation phenomena, resulting in large losses of transhydrogenase activity. The relation

  18. Organic hydrogels as potential sorbent materials for water purification (United States)

    Linardatos, George; Bekiari, Vlasoula; Bokias, George


    Hydrogels are three-dimensional, hydrophilic, polymeric networks capable to adsorb large amounts of water or biological fluids. The networks are composed of homopolymers or copolymers and are insoluble due to the presence of chemical or physical cross-links. Depending on the nature of the structural units, swelling or shrinking of these gels can be activated by several external stimuli, such as solvent, heat, pH, electric stimuli. As a consequence, these materials are attractive for several applications in a variety of fields: drug delivery, muscle mimetic soft linear actuators, hosts of nanoparticles and semiconductors, regenerative medicine etc. Of special interest is the application of hydrogels for water purification, since they can effectively adsorb several water soluble pollutants such as metal ions, inorganic or organic anions, organic dyestaff, etc. In the present work, anionic hydrogels bearing negatively charged -COO- groups were prepared and investigated. These are based on the anionic monomer sodium acrylate (ANa) and the nonionic one N,N-dimethylacrylamide (DMAM). A series of copolymeric hydrogels (P(DMAM-co-ANax) were synthesized. The molar content x of ANa units (expressing the molar charged content of the hydrogel) varies from 0 (nonionic poly(N,N-dimethylacrylamide), PDMAM, hydrogel) up to 1 (fully charged poly(sodium acrylate), PANa, hydrogel). The hydrogels were used to extract organic or inorganic solutes from water. Cationic and anionic model dyes, as well as multivalent inorganic ions, have been studied. It is found that cationic dyes are strongly adsorbed and retained by the hydrogels, while adsorbance of anionic dyes was negligible. Both maximum adsorption and equilibrium binding constant depend on the chemical structure of the dye, the presence of functional chemical groups and the hydrophobic-hydrophilic balance. In the case of metal cations, adsorption depends mostly on the charge of the cation. In addition, crucial factors controlling

  19. Purification and partial characterization of canine S100A12. (United States)

    Heilmann, Romy M; Suchodolski, Jan S; Steiner, Jörg M


    Canine S100A12 (cS100A12) is a calcium-binding protein of the S100 superfamily of EF-hand proteins, and its expression is restricted to neutrophils and monocytes. Interaction of S100A12 with the receptor for advanced glycation end products (RAGE) has been suggested to play a central role in inflammation. Moreover, S100A12 has been shown to represent a sensitive and specific marker for gastrointestinal inflammation in humans. Only human, porcine, bovine, and rabbit S100A12 have been purified to date, and an immunoassay for the quantification of S100A12 is available only for humans. Therefore, the aim of this study was to develop a protocol for the purification of S100A12 and to partially characterize this protein in the dog (Canis lupus familiaris) as a prelude to the development of an immunologic method for its detection and quantification in canine serum and fecal specimens. Leukocytes were isolated from canine whole blood by dextran sedimentation, and canine S100A12 was extracted from the cytosol fraction of these cells. Further purification of cS100A12 comprised of ammonium sulfate precipitation, hydrophobic interaction chromatography, and strong cation- and anion-exchange column chromatography. Canine S100A12 was successfully purified from canine whole blood. The relative molecular mass of the protein was estimated at 10,379.5 and isoelectric focusing revealed an isoelectric point of 6.0. The approximate specific absorbance of cS100A12 at 280 nm was determined to be 1.78 for a 1 mg/ml solution. The N-terminal AA sequence of the first 15 residues of cS100A12 was Thr-Lys-Leu-Glu-Asp-His-X-Glu-Gly-Ile-Val-Asp-Val-Phe-His, and revealed 100% identity with the predicted protein sequence available through the canine genome project. Sequence homology for the 14 N-terminal residues identified for cS100A12 with those of feline, bovine, porcine, and human S100A12 was 78.6%. We conclude that canine S100A12 can be successfully purified from canine whole blood using the

  20. Exploration of overloaded cation exchange chromatography for monoclonal antibody purification. (United States)

    Liu, Hui F; McCooey, Beth; Duarte, Tiago; Myers, Deanna E; Hudson, Terry; Amanullah, Ashraf; van Reis, Robert; Kelley, Brian D


    Cation exchange chromatography using conventional resins, having either diffusive or perfusive flow paths, operated in bind-elute mode has been commonly employed in monoclonal antibody (MAb) purification processes. In this study, the performance of diffusive and perfusive cation exchange resins (SP-Sepharose FF (SPSFF) and Poros 50HS) and a convective cation exchange membrane (Mustang S) and monolith (SO(3) Monolith) were compared. All matrices were utilized in an isocratic state under typical binding conditions with an antibody load of up to 1000 g/L of chromatographic matrix. The dynamic binding capacity of the cation exchange resins is typically below 100 g/L resin, so they were loaded beyond the point of anticipated MAb break through. All of the matrices performed similarly in that they effectively retained host cell protein and DNA during the loading and wash steps, while antibody flowed through each matrix after its dynamic binding capacity was reached. The matrices differed, though, in that conventional diffusive and perfusive chromatographic resins (SPSFF and Poros 50HS) demonstrated a higher binding capacity for high molecular weight species (HMW) than convective flow matrices (membrane and monolith); Poros 50HS displayed the highest HMW binding capacity. Further exploration of the conventional chromatographic resins in an isocratic overloaded mode demonstrated that the impurity binding capacity was well maintained on Poros 50HS, but not on SPSFF, when the operating flow rate was as high as 36 column volumes per hour. Host cell protein and HMW removal by Poros 50HS was affected by altering the loading conductivity. A higher percentage of host cell protein removal was achieved at a low conductivity of 3 mS/cm. HMW binding capacity was optimized at 5 mS/cm. Our data from runs on Poros 50HS resin also showed that leached protein A and cell culture additive such as gentamicin were able to be removed under the isocratic overloaded condition. Lastly, a MAb

  1. Tetanus toxoid purification: chromatographic procedures as an alternative to ammonium-sulphate precipitation. (United States)

    Stojićević, Ivana; Dimitrijević, Ljiljana; Dovezenski, Nebojša; Živković, Irena; Petrušić, Vladimir; Marinković, Emilija; Inić-Kanada, Aleksandra; Stojanović, Marijana


    Given an existing demand to establish a process of tetanus vaccine production in a way that allows its complete validation and standardization, this paper focuses on tetanus toxoid purification step. More precisely, we were looking at a possibility to replace the widely used ammonium-sulphate precipitation by a chromatographic method. Based on the tetanus toxin's biochemical characteristics, we have decided to examine the possibility of tetanus toxoid purification by hydrophobic chromatography, and by chromatographic techniques based on interaction with immobilized metal ions, i.e. chelating chromatography and immobilized metal affinity chromatography. We used samples obtained from differently fragmented crude tetanus toxins by formaldehyde treatment (assigned as TTd-A and TTd-B) as starting material for tetanus toxoid purification. Obtained results imply that purification of tetanus toxoid by hydrophobic chromatography represents a good alternative to ammonium-sulphate precipitation. Tetanus toxoid preparations obtained by hydrophobic chromatography were similar to those obtained by ammonium-sulphate precipitation in respect to yield, purity and immunogenicity. In addition, their immunogenicity was similar to standard tetanus toxoid preparation (NIBSC, Potters Bar, UK). Furthermore, the characteristics of crude tetanus toxin preparations had the lowest impact on the final purification product when hydrophobic chromatography was the applied method of tetanus toxoid purification. On the other hand, purifications of tetanus toxoid by chelating chromatography or immobilized metal affinity chromatography generally resulted in a very low yield due to not satisfactory tetanus toxoid binding to the column, and immunogenicity of the obtained tetanus toxoid-containing preparations was poor. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. Optimized Expression and Purification for High-Activity Preparations of Algal [FeFe]-Hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Yacoby, I.; Tegler, L. T.; Pochekailov, S.; Zhang, S.; King, P. W.


    Recombinant expression and purification of metallo-enzymes, including hydrogenases, at high-yields is challenging due to complex, and enzyme specific, post-translational maturation processes. Low fidelities of maturation result in preparations containing a significant fraction of inactive, apo-protein that are not suitable for biophysical or crystallographic studies. We describe the construction, overexpression and high-yield purification of a fusion protein consisting of the algal [2Fe2S]-ferredoxin PetF (Fd) and [FeFe]-hydrogenase HydA1. The maturation of Fd-HydA1 was optimized through improvements in culture conditions and media components used for expression. We also demonstrated that fusion of Fd to the N-terminus of HydA1, in comparison to the C-terminus, led to increased expression levels that were 4-fold higher. Together, these improvements led to enhanced HydA1 activity and improved yield after purification. The strong binding-affinity of Fd for DEAE allowed for two-step purification by ion exchange and StrepTactin affinity chromatography. In addition, the incorporation of a TEV protease site in the Fd-HydA1 linker allowed for the proteolytic removal of Fd after DEAE step, and purification of HydA1 alone by StrepTactin. In combination, this process resulted in HydA1 purification yields of 5 mg L{sup -1} of culture from E. coli with specific activities of 1000 U (U = 1 {micro}mol hydrogen evolved mg{sup -1} min{sup -1}). The [FeFe]-hydrogenases are highly efficient enzymes and their catalytic sites provide model structures for synthetic efforts to develop robust hydrogen activation catalysts. In order to characterize their structure-function properties in greater detail, and to use hydrogenases for biotechnological applications, reliable methods for rapid, high-yield expression and purification are required.

  3. Prediction of purification of biopharmeceuticals with molecular dynamics (United States)

    Ustach, Vincent; Faller, Roland

    Purification of biopharmeceuticals remains the most expensive part of protein-based drug production. In ion exchange chromatography (IEX), prediction of the elution ionic strength of host cell and target proteins has the potential to reduce the parameter space for scale-up of protein production. The complex shape and charge distribution of proteins and pores complicates predictions of the interactions in these systems. All-atom molecular dynamics methods are beyond the scope of computational limits for mass transport regimes. We present a coarse-grained model for proteins for prediction of elution pH and ionic strength. By extending the raspberry model for colloid particles to surface shapes and charge distributions of proteins, we can reproduce the behavior of proteins in IEX. The average charge states of titratatable amino acid residues at relevant pH values are determined by extrapolation from all-atom molecular dynamics at pH 7. The pH specific all-atom electrostatic field is then mapped onto the coarse-grained surface beads of the raspberry particle. The hydrodynamics are reproduced with the lattice-Boltzmann scheme. This combination of methods allows very long simulation times. The model is being validated for known elution procedures by comparing the data with experiments. Defense Threat Reduction Agency (Grant Number HDTRA1-15-1-0054).

  4. Extraction purification and characterization of trypsin inhibitors from Andean seeds

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    Patricio Castillo


    Full Text Available This work established the conditions of covalent immobilization of trypsin on a Sepharose matrix, which could be applied for the purification of trypsin inhibitors. The higher values of retention of enzymatic activity and immobilized enzymatic activity were obtained with a Sepharose 6B-CL matrix, at room temperature, a pH value of 10.5, an enzymatic load of 25 mg/mL, and a minimum immobilization time of 12 hours, in order to obtain a stable immobilization. The most active trypsin inhibitors were selected through the comparison of, extracts obtained from the seeds of amaranth (Amaranthus caudatus L., pea (Pisum sativum, lupine or “chocho” (Lupinus mutabilis, bean (Phaseolus vulgaris and “sangorache” (Amaranthus hybridus L.. The inhibitors were partially purified using centrifugal ultrafiltration, heat treatment, and TCA precipitation. The permeated and retained fractions of “sangorache” were selected as the most active trypsin inhibitors, and they were selectively purified using affinity chromatography in a Trypsin - Glyoxyl - Sepharose 6B-CL matrix. The kinetic characterization showed the presence of two inhibitors; the first one corresponded to a competitive inhibitor, while the second one behaved as a mixed inhibitor.

  5. Purification and proteomics of pathogen-modified vacuoles and membranes. (United States)

    Herweg, Jo-Ana; Hansmeier, Nicole; Otto, Andreas; Geffken, Anna C; Subbarayal, Prema; Prusty, Bhupesh K; Becher, Dörte; Hensel, Michael; Schaible, Ulrich E; Rudel, Thomas; Hilbi, Hubert


    Certain pathogenic bacteria adopt an intracellular lifestyle and proliferate in eukaryotic host cells. The intracellular niche protects the bacteria from cellular and humoral components of the mammalian immune system, and at the same time, allows the bacteria to gain access to otherwise restricted nutrient sources. Yet, intracellular protection and access to nutrients comes with a price, i.e., the bacteria need to overcome cell-autonomous defense mechanisms, such as the bactericidal endocytic pathway. While a few bacteria rupture the early phagosome and escape into the host cytoplasm, most intracellular pathogens form a distinct, degradation-resistant and replication-permissive membranous compartment. Intracellular bacteria that form unique pathogen vacuoles include Legionella, Mycobacterium, Chlamydia, Simkania, and Salmonella species. In order to understand the formation of these pathogen niches on a global scale and in a comprehensive and quantitative manner, an inventory of compartment-associated host factors is required. To this end, the intact pathogen compartments need to be isolated, purified and biochemically characterized. Here, we review recent progress on the isolation and purification of pathogen-modified vacuoles and membranes, as well as their proteomic characterization by mass spectrometry and different validation approaches. These studies provide the basis for further investigations on the specific mechanisms of pathogen-driven compartment formation.

  6. Recombinant allergen Lol p II: expression, purification and characterization. (United States)

    Tamborini, E; Brandazza, A; De Lalla, C; Musco, G; Siccardi, A G; Arosio, P; Sidoli, A


    Pollen from perennial rye grass (Lolium perenne) is a major cause of type I allergies worldwide. It contains complex mixtures of proteins, among which Lol p II is a major allergen. Previously, we have reported the cloning and sequencing of Lol p II and its expression in fusion with the heavy chain of human ferritin as carrier polypeptide (Sidoli et al., 1993, J. biol. Chem. 268, 21819-21825). Here, we describe the expression, purification and characterization of a recombinant Lol p II overproduced as a non-fusion protein in the periplasm of E. coli. The recombinant allergen was expressed in high yields and was easily purified in milligram amounts. It competed with the natural Lol p II for binding to specific IgE, and it induced allergic responses in skin prick tests, indicating to be immunologically analogous to the natural protein. Biochemical analyses indicate that recombinant Lol p II is a highly stable and soluble monomeric molecule which behaves like a small globular protein.

  7. Simplified, enhanced protein purification using an inducible, autoprocessing enzyme tag.

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    Aimee Shen


    Full Text Available We introduce a new method for purifying recombinant proteins expressed in bacteria using a highly specific, inducible, self-cleaving protease tag. This tag is comprised of the Vibrio cholerae MARTX toxin cysteine protease domain (CPD, an autoprocessing enzyme that cleaves exclusively after a leucine residue within the target protein-CPD junction. Importantly, V. cholerae CPD is specifically activated by inositol hexakisphosphate (InsP(6, a eukaryotic-specific small molecule that is absent from the bacterial cytosol. As a result, when His(6-tagged CPD is fused to the C-terminus of target proteins and expressed in Escherichia coli, the full-length fusion protein can be purified from bacterial lysates using metal ion affinity chromatography. Subsequent addition of InsP(6 to the immobilized fusion protein induces CPD-mediated cleavage at the target protein-CPD junction, releasing untagged target protein into the supernatant. This method condenses affinity chromatography and fusion tag cleavage into a single step, obviating the need for exogenous protease addition to remove the fusion tag(s and increasing the efficiency of tag separation. Furthermore, in addition to being timesaving, versatile, and inexpensive, our results indicate that the CPD purification system can enhance the expression, integrity, and solubility of intractable proteins from diverse organisms.

  8. EB technology for the purification of flue gases

    International Nuclear Information System (INIS)

    Kojima, Takuji


    Sulfur oxides and nitrogen oxides in flue gas from coal-combustion boilers in power plants, dioxins in flue gas from municipal waste incineration facilities and toxic volatile organic compounds (VOCs) in off-gas from painting or cleaning factories are among air pollutants for which emission is regulated by a law in Japan. Electron beam is the effective and easy controllable radiation source for treatment of these flue gases. This report describes outline of the results so far obtained at JAERI on electron beam treatment of flue gas. The removal performance higher than 90% at 10 kGy for flue gas containing 800 ppm SOx and 225 ppm NOx were achieved and being applied to real-scale power plants in Poland and China with expectation of cost reduction of 20% compared to conventional plants. Decomposition of dioxins in flue gas from solid waste incinerators is another project. Using an accelerator of 300 keV and 40 mA for treatment of real incineration gas at 200degC, we obtain 90% decomposition of dioxins at 15 kGy irradiation. Expansion of these flue gas purification technologies combined with low-energy electron accelerators is expected. (S. Ohno)

  9. Advanced Water Purification System for In Situ Resource Utilization Project (United States)

    Anthony, Stephen M.


    A main goal in the field of In Situ Resource Utilization is to develop technologies that produce oxygen from regolith to provide consumables to an extratrrestrial outpost. The processes developed reduce metal oxides in the regolith to produce water, which is then electrolyzed to produce oxygen. Hydrochloric and hydrofluoric acids are byproducts of the reduction processes, which must be removed to meet electrolysis purity standards. We previously characterized Nation, a highly water selective polymeric proton-exchange membrane, as a filtrtion material to recover pure water from the contaminated solution. While the membranes successfully removed both acid contaminants, the removal efficiency of and water flow rate through the membranes were not sufficient to produce large volumes of electrolysis-grade water. In the present study, we investigated electrodialysis as a potential acid removable technique. Our studies have show a rapid and significant reduction in chloride and fluoride concentrations in the feed solution, while generating a relatively small volume of concentrated waste water. Electrodialysis has shown significant promise as the primary separation technique in ISRU water purification processes.

  10. Extraction, Purification, and NMR Analysis of Terpenes from Brown Algae. (United States)

    Gaysinski, Marc; Ortalo-Magné, Annick; Thomas, Olivier P; Culioli, Gérald


    Algal terpenes constitute a wide and well-documented group of marine natural products with structures differing from their terrestrial plant biosynthetic analogues. Amongst macroalgae, brown seaweeds are considered as one of the richest source of biologically and ecologically relevant terpenoids. These metabolites, mostly encountered in algae of the class Phaeophyceae, are mainly diterpenes and meroditerpenes (metabolites of mixed biogenesis characterized by a toluquinol or a toluquinone nucleus linked to a diterpene moiety).In this chapter, we describe analytical processes commonly employed for the isolation and structural characterization of the main terpenoid constituents obtained from organic extracts of brown algae. The successive steps include (1) extraction of lipidic content from algal samples; (2) purification of terpenes by column chromatography and semi-preparative high-performance liquid chromatography; and (3) structure elucidation of the isolated terpenes by means of 1D and 2D nuclear magnetic resonance (NMR). More precisely, we propose a representative methodology which allows the isolation and structural determination of the monocyclic meroditerpene methoxybifurcarenone (MBFC) from the Mediterranean brown alga Cystoseira amentacea var. stricta. This methodology has a large field of applications and can then be extended to terpenes isolated from other species of the family Sargassaceae.

  11. Carbonic anhydrase from Apis mellifera: purification and inhibition by pesticides. (United States)

    Soydan, Ercan; Güler, Ahmet; Bıyık, Selim; Şentürk, Murat; Supuran, Claudiu T; Ekinci, Deniz


    Carbonic anhydrase (CA) enzymes have been shown to play an important role in ion transport and in pH regulation in several organisms. Despite this information and the wealth of knowledge regarding the significance of CA enzymes, few studies have been reported about bee CA enzymes and the hazardous effects of chemicals. Using Apis mellifera as a model, this study aimed to determine the risk of pesticides on Apis mellifera Carbonic anhydrase enzyme (Am CA). CA was initially purified from Apis mellifera spermatheca for the first time in the literature. The enzyme was purified with an overall purification of ∼35-fold with a molecular weight of ∼32 kDa. The enzyme was then exposed to pesticides, including tebuconazole, propoxur, carbaryl, carbofuran, simazine and atrazine. The six pesticides dose-dependently inhibited in vitro AmCA activity at low micromolar concentrations. IC 50 values for the pesticides were 0.0030, 0.0321, 0.0031, 0.0087, 0.0273 and 0.0165 μM, respectively. The AmCA inhibition mechanism of these compounds is unknown at this moment.

  12. Cloning, expression, purification and characterization of tryptophan hydroxylase variants

    DEFF Research Database (Denmark)

    Boesen, Jane

    in the anion exchange, indicating that the protein still exists in different oligomer forms. This was also observed in the gel filtration. Variants of both hTPH1 and hTPH2 containing the regulatory domain or parts of it were constructed and tested for expression in Escherichia coli as well as solubility....... It was observed that changes in the amino acid sequence of the regulatory domain by point mutations or truncations in the N-terminal had a huge impact on the solubility of the protein and caused the protein to be insoluble. The regulatory domain of human TPH1 (rhTPH1), and two fusion proteins of rhTPH1 fused...... to the green fluorescent protein (GFP) in the C-terminal and the glutathione S-transferase (GST) in the N-terminal, respectively, were expressed in a soluble form. The purification trials of the variants containing the regulatory domain showed that a high salt concentration was necessary to stabilize...

  13. Expression, Purification, and Characterisation of Dehydroquinate Synthase from Pyrococcus furiosus

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    Leonardo Negron


    Full Text Available Dehydroquinate synthase (DHQS catalyses the second step of the shikimate pathway to aromatic compounds. DHQS from the archaeal hyperthermophile Pyrococcus furiosus was insoluble when expressed in Escherichia coli but was partially solubilised when KCl was included in the cell lysis buffer. A purification procedure was developed, involving lysis by sonication at 30∘C followed by a heat treatment at 70∘C and anion exchange chromatography. Purified recombinant P. furiosus DHQS is a dimer with a subunit Mr of 37,397 (determined by electrospray ionisation mass spectrometry and is active over broad pH and temperature ranges. The kinetic parameters are KM (3-deoxy-D-arabino-heptulosonate 7-phosphate 3.7 μM and kcat 3.0 sec-1 at 60∘C and pH 6.8. EDTA inactivates the enzyme, and enzyme activity is restored by several divalent metal ions including (in order of decreasing effectiveness Cd2+, Co2+, Zn2+, and Mn2+. High activity of a DHQS in the presence of Cd2+ has not been reported for enzymes from other sources, and may be related to the bioavailability of Cd2+ for P. furiosus. This study is the first biochemical characterisation of a DHQS from a thermophilic source. Furthermore, the characterisation of this hyperthermophilic enzyme was carried out at elevated temperatures using an enzyme-coupled assay.

  14. Expression and purification of the Sgm protein from E. coli

    Directory of Open Access Journals (Sweden)



    Full Text Available The sgm gene from Micromonospora zionensis, the producer of the aminoglycoside antibiotic G-52, encodes for Sgm methylasewhich modifies the target site on 16S rRNA and thus protects the producer against its own toxic product. The sgm gene wasmodified by polymerase chain reaction (PCR and cloned in the QIAexpress pQE-30 vector in order to make a construct that places the (His6 tag at the N-terminus of the protein. The resulting expression construct was transformed in the E. coli strain NM522 and the functional activity of the Sgm-His fusion protein was confirmed in vivo. Purification of the (His6-tagged Sgm protein by Ni-NTA affinity chromatography was performed under native conditions and the protein was detected on a sodium dodecyl sulfate polyacrylamide gel. Sgm methylase was purified to homogeneity > 95 %. Polyclonal antibodies raised to purified (His6-tagged Sgm protein were used to identify this protein byWestern blot analysis.

  15. [Purification and properties of NAG A from human kidney]. (United States)

    Yoshida, K


    In the present paper, we have reported the purification procedures of N-acetyl-beta, D-glucosaminidase (NAG) A from human renal tissue as well as the enzymatic properties of NAG A. NAG A was purified to homogeneity by gel filtration methods using Sephacry S-400 and S-200, followed by affinity chromatography with TSK DEAE 5-PW. The final activity of the enzyme was 1001 U/ml protein which was 506.6-fold that of the crude extract (supernatant of 20,000 x G of the homogenate). The molecular weight of NAG A was 140 kDa, consisting of two subunits of 30 kDa and 57 kDa. The isoelectric point of the enzyme was 5.60. The optimal pH of the enzyme was between 4.7 and 4.9. The Km value of the enzyme for sodio-m-cresol sulfophtaleinyl-N-acetyl-beta, D-glucosaminide was found 0.177 x 10(-3) mol/l. Lectin affinity chromatographies using concanavalin A and wheat germagglutinin have demonstrated that major sugar-chains of the enzyme were the high mannose type and hybrid type with a fucose residue, and that a small amount of the complex type was contained.

  16. Purification and characterization of a prolyl aminopeptidase from Debaryomyces hansenii. (United States)

    Bolumar, Tomás; Sanz, Yolanda; Aristoy, M-Concepción; Toldrá, Fidel


    A prolyl aminopeptidase (PAP) (EC was isolated from the cell extract of Debaryomyces hansenii CECT12487. The enzyme was purified by selective fractionation with protamine and ammonium sulfate, followed by two chromatography steps, which included gel filtration and anion-exchange chromatography. The PAP was purified 248-fold, with a recovery yield of 1.4%. The enzyme was active in a broad pH range (from 5 to 9.5), with pH and temperature optima at 7.5 and 45 degrees C. The molecular mass was estimated to be around 370 kDa. The presence of inhibitors of serine and aspartic proteases, bestatin, puromycin, reducing agents, chelating agents, and different cations did not have any effect on the enzyme activity. Only iodoacetate, p-chloromercuribenzoic acid, and Hg(2+), which are inhibitors of cysteine proteases, markedly reduced the enzyme activity. The K(m) for proline-7-amido-4-methylcoumarin was 40 micro M. The enzyme exclusively hydrolyzed N-terminal-proline-containing substrates. This is the first report on the identification and purification of this type of aminopeptidase in yeast, which may contribute to the scarce knowledge about D. hansenii proteases and their possible roles in meat fermentation.

  17. Synthesis, isolation and purification of [11C]-choline

    Directory of Open Access Journals (Sweden)

    Marcin Szydło


    Full Text Available [11C]-choline is an effective PET tracer used for imaging of neoplastic lesions and metastases of the prostate cancer. However, its production can be a challenge for manufacturers, as it has not yet been described in Polish or European pharmacopoeia. In this study the technical aspects of [11C]-choline production are described and detailed process parameters are provided. The quality control procedures for releasing [11C]-choline as solutio iniectabilis are also presented. The purity and quality of the radiopharmaceutical obtained according to the proposed method were find to be high enough to safely administrate the radiopharmaceutical to patients. Application of an automated synthesizer makes it possible to carry out the entire process of [11C]-choline production, isolation and purification within 20 minutes. It is crucial to maintain all aspects of the process as short as possible, since the decay half-time of carbon-11 is 20.4 minutes. The resulting radiopharmaceutical is sterile and pyrogen-free and of a high chemical, radiochemical, and radionuclide purity proved by chromatographic techniques. The yield of the process is up to 20%. [11C]-choline PET scanning can be used as accurate and effective diagnostic tool in all centers equipped with [11C]-target containing cyclotron.

  18. Purification and characterization of tyrosinase from walnut leaves (Juglans regia). (United States)

    Zekiri, Florime; Molitor, Christian; Mauracher, Stephan G; Michael, Claudia; Mayer, Rupert L; Gerner, Christopher; Rompel, Annette


    Polyphenol oxidase (PPO) is a type-3 copper enzyme catalyzing the oxidation of phenolic compounds to their quinone derivates, which are further converted to melanin, a ubiquitous pigment in living organisms. In this study a plant originated tyrosinase was isolated from walnut leaves (Juglans regia) and biochemically characterized. It was possible to isolate and purify the enzyme by means of an aqueous two-phase extraction method followed by chromatographic purification and identification. Interestingly, the enzyme showed a rather high monophenolase activity considering that the main part of plant PPOs with some exceptions solely possess diphenolase activity. The average molecular mass of 39,047 Da (Asp(101)→Arg(445)) was determined very accurately by high resolution mass spectrometry. This proteolytically activated tyrosinase species was identified as a polyphenol oxidase corresponding to the known jrPPO1 sequence by peptide sequencing applying nanoUHPLC-ESI-MS/MS. The polypeptide backbone with sequence coverage of 96% was determined to start from Asp(101) and not to exceed Arg(445). Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  19. Purification and Partial Characterization of Trypanosoma cruzi Triosephosphate Isomerase

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    Bourguignon SC


    Full Text Available The enzyme triosephosphate isomerase (TPI, EC was purified from extracts of epimastigote forms of Trypanosoma cruzi. The purification steps included: hydrophobic interaction chromatography on phenyl-Sepharose, CM-Sepharose, and high performance liquid gel filtration chromatography. The CM-Sepharose material contained two bands (27 and 25 kDa with similar isoelectric points (pI 9.3-9.5 which could be separated by gel filtration in high performance liquid chromatography. Polyclonal antibodies raised against the porcine TPI detected one single polypeptide on western blot with a molecular weight (27 kDa identical to that purified from T. cruzi. These antibodies also recognized only one band of identical molecular weight in western blots of several other trypanosomatids (Blastocrithidia culicis, Crithidia desouzai, Phytomonas serpens, Herpertomonas samuelpessoai. The presence of only one enzymatic form of TPI in T. cruzi epimastigotes was confirmed by agarose gel activity assay and its localization was established by immunocytochemical analysis. The T. cruzi purified TPI (as well as other trypanosomatid' TPIs is a dimeric protein, composed of two identical subunits with an approximate mw of 27,000 and it is resolved on two dimensional gel electrophoresis with a pI of 9.3. Sequence analysis of the N-terminal portion of the 27 kDa protein revealed a high homology to Leishmania mexicana and T. brucei proteins

  20. Development of composite metallic membranes for hydrogen purification

    International Nuclear Information System (INIS)

    Gaillard, F.


    Fuel cells are able to convert chemical energy into electric power. There are different types of cells; the best for automotive applications are Proton Exchange Membrane Fuel Cells. But, these systems need hydrogen of high purity. However, fuel reforming generates a mixture of gases, from which hydrogen has to be extracted before supplying the electrochemical cell. The best way for the purification of hydrogen is the membrane separation technology. Palladium is selectively permeable to hydrogen and this is the reason why this metal is largely used for the membrane development. This work deals with the development of hydrogen-selective membranes by deposition of a thin film of palladium onto a porous mechanical support. For this, we have used the electroless plating technique: a palladium salt and a reducing agent are mixed and the deposition takes place onto the catalytic surface of the substrate. After bibliographic investigations, experimental studies have been performed first with a dense metallic substrate in order to better understand the different parameters controlling the deposition. First of all, potentiometric measurements have been carried out to follow the electrochemical reactions in the bath. Then, kinetic measurements of the coating thickness have been recorded to understand the effect of the bath conditions on the yield and the adhesion of the film. Finally, the electroless plating method has been applied to deposit palladium membranes onto porous stainless steel substrates. After optimisation, the resulting membranes were tested for their hydrogen permeation properties. (author)