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Sample records for thermophilum xylanase 11-a

  1. Utilization of deoiled Jatropha curcas seed cake for production of xylanase from thermophilic Scytalidium thermophilum.

    Science.gov (United States)

    Joshi, Chetna; Khare, S K

    2011-01-01

    Jatropha curcas is a major biodiesel crop. Large amount of deoiled cake is generated as by-product during biodiesel production from its seeds. Deoiled J. curcas seed cake was assessed as substrate for the production of xylanase from thermophilic fungus Scytalidium thermophilum by solid-state fermentation. The seed cake was efficiently utilized by S. thermophilum for its growth during which it produced good amount of heat stable extracellular xylanase. The solid-state fermentation conditions were optimized for maximum xylanase production. Under the optimized conditions viz. deoiled seed cake supplemented with 1% oat-spelt xylan, adjusted to pH 9.0, moisture content 1:3 w/v, inoculated with 1×10(6) spores per 5 g cake and incubated at 45 °C, 1455 U xylanase/g deoiled seed cake was obtained. The xylanase was useful in biobleaching of paper pulp. Solid-state fermentation of deoiled cake appears a potentially viable approach for its effective utilization. Copyright © 2010 Elsevier Ltd. All rights reserved.

  2. Cloning and expression of chaetomium thermophilum xylanase 11-A

    International Nuclear Information System (INIS)

    Andleeb, S.; Latif, F.; Afzal, S.; Mukhtar, Z.; Mansoor, S.; Rajoka, I.

    2008-01-01

    The various thermophilic fungi like Chaetomium thermophile has potential to secrete xylanase and cellulase enzymes. In the present study eukaryotic expression system of Pichia pastoris (yeast) was used to express xylanase gene. The xylanase (Xyn 11-A) gene was isolated from C. thermophile strain NIBGE-1. Primers were designed to amplify the gene, ligated into P. pastoris pPIC3.5K vector, the resultant recombinant clone pSSZ810 was transformed into the genome of P. pastoris GS115 strain through electroporation. Transformants were selected on yeast peptone dextrose medium (YPD) plates containing antibiotic geneticin (100 mg/ml) upto final concentration of 0.75 mg/ml. The maximum activity of xylanase 2.04 U/ml after incubation of 2 hours at 50 degree C was observed in the presence of 100% methanol inducer upto final concentration of 30 macro L (0.5%) as compared to control. HPLC analysis represented high peak of xylose as compared to control. SDS-PAGE indicated approx. 28 kDa protein of expressed xylanase gene. (author)

  3. Heterologous expression of chaetomium thermophilum xylanase 11-a (ctx 11-a) gene

    International Nuclear Information System (INIS)

    Wajid, S.; Shahid, S.; Mukhtar, Z.; Mansoor, S.

    2009-01-01

    Chaetomium has a potential source of xylanase and cellulase enzymes, both of which are required in the treatment of fibre in the poultry feed. The titre of the enzymes needs to be enhanced by using recombinant DNA technology for fulfilling the requirement of the industries. Efforts are made to construct prokaryotic and eukaryotic expression cassettes that can be cloned under specific strong promoters i.e., T7 and AOX1, respectively, and the enhancer elements to get the maximum gene expression. In the present study BL21 E. coli and GS115 Pichia pastoris strains are used as model organisms to express the CtX 11-A gene in the presence of 1 mM IPTG and 100% methanol upto final concentration of 0.5. In case of BL21 expression, the maximum xylanase activity was observed after 1.5 h in the presence of 1% xylose, which was 2.302 U/ml and after 7 h in the presence of 0.5% lactose, was 1.708 U/ml. However, in Pichia pastoris the maximum production of xylanase was 2.904 and 0.006 U/ml as compared to control 0.484 and 0.06 U/ml, respectively. (author)

  4. Cloning and expression of chaetomium thermophilum xylanase 11-A gene in prokaryote

    International Nuclear Information System (INIS)

    Wajid, S.; Latif, F.; Afzal, S.; Rajoka, I.

    2008-01-01

    The xylanase gene was cloned into pET32a(+) and expressed in E. coli BL21 under T7 promotor alongwith fusion protein. The SDS-PAGE and western blot analysis showed a protein of 42 kDa. The best expression of xylanase enzyme was found by using xylose as carbon source and lactose as an inducer. The maximum activity of xylanase expressed in E. coli was 6.02 U/mL in the presence of 2% xylose in DS medium. The activity of recombinant xylanase was observed on 1% xylan LB agar plates, showed halos of xylan clearance when lactose was used as an inducer. (author)

  5. Thermophilic and alkalophilic xylanases from several Dictyoglomus isolates

    Energy Technology Data Exchange (ETDEWEB)

    Mathrani, I M; Ahring, B K [Technical Univ. of Denmark, Lyngby (Denmark). Anaerobic Microbiology/Biotechnology Group

    1992-10-01

    Supernatant xylanases from three thermophilic and strictly anaerobic Dictyoglomus strains isolated from very different environments were examined: The type species, D. thermophilum[sup T], from a hot-spring in Japan; strain B1, a recently described strictly xylanutilizing Dictyoglomus from a paper-pulp factory in Finland; and strain B4a, isolated from a thermal pool on Iceland. The highest enzymatic activity observed from batch-culture supernatant with 4 g l[sup -1] of beech xylan as growth substrate was 3.8x10[sup -6] kat l[sup -1]. The K[sub m] for the xylanases of strain B1 was 4.7 g beech xylan l[sup -1]. The xylanases of all the isolates had a broad range of activity with respect to pH, showing good activity from pH 5.5 to near 9.0. The xylanases from the three isolates had a very high temperature optimum of 80deg C, maximum temperature for extended activity between 80 and 90deg C, and a thermal half-life of over 1 h at 90deg C for strain B1. The application of thermophilic alkalophilic xylanases to paper pulping was discussed. (orig.).

  6. Autohydrolysis of plant xylans by apoplastic expression of thermophilic bacterial endo-xylanases

    DEFF Research Database (Denmark)

    Borkhardt, Bernhard; Harholt, Jesper; Ulvskov, Peter Bjarne

    2010-01-01

    The genes encoding the two endo-xylanases XynA and XynB from the thermophilic bacterium Dictyoglomus thermophilum were codon optimized for expression in plants. Both xylanases were designed to be constitutively expressed under the control of the CaMV 35S promoter and targeted to the apoplast....... Transient expression in tobacco and stable expression in transgenic Arabidopsis showed that both enzymes were expressed in an active form with temperature optima at 85 °C. Transgenic Arabidopsis accumulating heterologous endo-xylanases appeared phenotypically normal and were fully fertile. The highest...... xylanase activity in Arabidopsis was found in dry stems indicating that the enzymes were not degraded during stem senescence. High levels of enzyme activity were maintained in cell-free extracts from dry transgenic stems during incubation at 85 °C for 24 h. Analysis of cell wall polysaccharides after heat...

  7. The Botrytis cinerea xylanase Xyn11A contributes to virulence with its necrotizing activity, not with its catalytic activity

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    González Celedonio

    2010-02-01

    Full Text Available Abstract Background The Botrytis cinerea xylanase Xyn11A has been previously shown to be required for full virulence of this organism despite its poor contribution to the secreted xylanase activity and the low xylan content of B. cinerea hosts. Intriguingly, xylanases from other fungi have been shown to have the property, independent of the xylan degrading activity, to induce necrosis when applied to plant tissues, so we decided to test the hypothesis that secreted Xyn11A contributes to virulence by promoting the necrosis of the plant tissue surrounding the infection, therefore facilitating the growth of this necrotroph. Results We show here that Xyn11A has necrotizing activity on plants and that this capacity is conserved in site-directed mutants of the protein lacking the catalytic activity. Besides, Xyn11A contributes to the infection process with the necrotizing and not with the xylan hydrolyzing activity, as the catalytically-impaired Xyn11A variants were able to complement the lower virulence of the xyn11A mutant. The necrotizing activity was mapped to a 30-amino acids peptide in the protein surface, and this region was also shown to mediate binding to tobacco spheroplasts by itself. Conclusions The main contribution of the xylanase Xyn11A to the infection process of B. cinerea is to induce necrosis of the infected plant tissue. A conserved 30-amino acids region on the enzyme surface, away from the xylanase active site, is responsible for this effect and mediates binding to plant cells.

  8. High genetic diversity and different distributions of glycosyl hydrolase family 10 and 11 xylanases in the goat rumen.

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    Guozeng Wang

    Full Text Available BACKGROUND: The rumen harbors a complex microbial ecosystem for efficient hydrolysis of plant polysaccharides which are the main constituent of the diet. Xylanase is crucial for hemicellulose hydrolysis and plays an important role in the plant cell wall degradation. Xylanases of ruminal strains were widely studied, but few studies have focused on their diversity in rumen microenvironment. METHODOLOGY/PRINCIPAL FINDINGS: We explored the genetic diversity of xylanases belonging to two major glycosyl hydrolase families (GH 10 and 11 in goat rumen contents by analyzing the amplicons generated with two degenerate primer sets. Fifty-two distinct GH 10 and 35 GH 11 xylanase gene fragments (similarity <95% were retrieved, and most had low identities with known sequences. Based on phylogenetic analysis, all GH 10 xylanase sequences fell into seven clusters, and 88.5% of them were related to xylanases from Bacteroidetes. Five clusters of GH 11 xylanase sequences were identified. Of these, 85.7% were related to xylanases from Firmicutes, and 14.3% were related to those of rumen fungi. Two full-length xylanase genes (one for each family were directly cloned and expressed in Escherichia coli. Both the recombinant enzymes showed substantial xylanase activity, and were purified and characterized. Combined with the results of sheep rumen, Bacteroidetes and Firmicutes are the two major phyla of xylan-degrading microorganisms in rumen, which is distinct from the representatives of other environments such as soil and termite hindgut, suggesting that xylan-degrading microorganisms are environment specific. CONCLUSION/SIGNIFICANCE: The numerous new xylanase genes suggested the functional diversity of xylanase in the rumen microenvironment which may have great potential applications in industry and agriculture. The phylogenetic diversity and different distributions of xylanase genes will help us understand their roles in plant cell wall degradation in the rumen

  9. Kinetics and substrate selectivity of a Triticum aestivum xylanase inhibitor (TAXI) resistant D11F/R122D variant of Bacillus subtilis XynA xylanase

    DEFF Research Database (Denmark)

    Rasmussen, Louise Enggaard; Sørensen, Jens F.; Meyer, Anne S.

    2010-01-01

    This study examined the kinetics and substrate selectivity of a GH11 Bacillus subtilis XynA xylanase (BsX) sensitive to inhibition by TAXI and an engineered variant, which is much less inhibited by TAXI (BsX(mut)). The main purpose of the work was to elucidate any influence of the structural point...

  10. Classification of a moderately oxygen-tolerant isolate from baby faeces as Bifidobacterium thermophilum

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    Fliss Ismaïl

    2007-08-01

    Full Text Available Abstract Background Bifidobacteria are found at varying prevalence in human microbiota and seem to play an important role in the human gastrointestinal tract (GIT. Bifidobacteria are highly adapted to the human GIT which is reflected in the genome sequence of a Bifidobacterim longum isolate. The competitiveness against other bacteria is not fully understood yet but may be related to the production of antimicrobial compounds such as bacteriocins. In a previous study, 34 Bifidobacterium isolates have been isolated from baby faeces among which six showed proteinaceous antilisterial activity against Listeria monocytogenes. In this study, one of these isolates, RBL67, was further identified and characterized. Results Bifidobacterium isolate RBL67 was classified and characterized using a polyphasic approach. RBL67 was classified as Bifidobacterium thermophilum based on phenotypic and DNA-DNA hybridization characteristics, although 16S rDNA analyses and partial groEL sequences showed higher homology with B. thermacidophilum subsp. porcinum and B. thermacidophilum subsp. thermacidophilum, respectively. RBL67 was moderately oxygen-tolerant and was able to grow at pH 4 and at a temperature of 47°C. Conclusion In order to assign RBL67 to a species, a polyphasic approach was used. This resulted in the classification of RBL67 as a Bifidobacterium thermophilum strain. To our knowledge, this is the first report about B. thermophilum isolated from baby faeces since the B. thermophilum strains were related to ruminants and swine faeces before. B. thermophilum was previously only isolated from animal sources and was therefore suggested to be used as differential species between animal and human contamination. Our findings may disapprove this suggestion and further studies are now conducted to determine whether B. thermophilum is distributed broader in human faeces. Furthermore, the postulated differentiation between human and animal strains by growth above 45

  11. Engineering improved thermostability of the GH11 xylanase from Neocallimastix patriciarum via computational library design.

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    Bu, Yifan; Cui, Yinglu; Peng, Ying; Hu, Meirong; Tian, Yu'e; Tao, Yong; Wu, Bian

    2018-04-01

    Xylanases, which cleave the β-1,4-glycosidic bond between xylose residues to release xylooligosaccharides (XOS), are widely used as food additives, animal feeds, and pulp bleaching agents. However, the thermally unstable nature of xylanases would hamper their industrial application. In this study, we used in silico design in a glycoside hydrolase family (GH) 11 xylanase to stabilize the enzyme. A combination of the best mutations increased the apparent melting temperature by 14 °C and significantly enhanced thermostability and thermoactivation. The variant also showed an upward-shifted optimal temperature for catalysis without compromising its activity at low temperatures. Moreover, a 10-fold higher XOS production yield was obtained at 70 °C, which compensated the low yield obtained with the wild-type enzyme. Collectively, the variant constructed by the computational strategy can be used as an efficient biocatalyst for XOS production at industrially viable conditions.

  12. Hydrolysis of Wheat Arabinoxylan by Two Acetyl Xylan Esterases from Chaetomium thermophilum

    DEFF Research Database (Denmark)

    Tong, Xiaoxue; Lange, Lene; Grell, Morten Nedergaard

    2015-01-01

    The thermophilic filamentous ascomycete Chaetomium thermophilum produces functionally diverse hemicellulases when grown on hemicellulose as carbon source. Acetyl xylan esterase (EC 3.1.1.72) is an important accessory enzyme in hemicellulose biodegradation. Although the genome of C. thermophilum has...

  13. Heliotropium thermophilum (Boraginaceae), a new taxon from SW Anatolia, Turkey

    DEFF Research Database (Denmark)

    Tan, Kit; Celik, Ali; Gemici, Yusuf

    2008-01-01

    Heliotropium thermophilum Kit Tan, A. Çelik & Y. Gemici (Boraginaceae), is described as a species new to science and illustrated. Its diploid chromosome number of 2n = 16 is a first report. It is restricted to the province of Aydin bordering on Denizli in SW Anatolia and is of interest on account...

  14. Microbial xylanases: engineering, production and industrial applications.

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    Juturu, Veeresh; Wu, Jin Chuan

    2012-01-01

    Enzymatic depolymerization of hemicellulose to monomer sugars needs the synergistic action of multiple enzymes, among them endo-xylanases (EC 3.2.1.8) and β-xylosidases (EC 3.2.1.37) (collectively xylanases) play a vital role in depolymerizing xylan, the major component of hemicellulose. Recent developments in recombinant protein engineering have paved the way for engineering and expressing xylanases in both heterologous and homologous hosts. Functional expression of endo-xylanases has been successful in many hosts including bacteria, yeasts, fungi and plants with yeasts being the most promising expression systems. Functional expression of β-xylosidases is more challenging possibly due to their more complicated structures. The structures of endo-xylanases of glycoside hydrolase families 10 and 11 have been well elucidated. Family F/10 endo-xylanases are composed of a cellulose-binding domain and a catalytic domain connected by a linker peptide with a (β/α)8 fold TIM barrel. Family G/11 endo-xylanases have a β-jelly roll structure and are thought to be able to pass through the pores of hemicellulose network owing to their smaller molecular sizes. The structure of a β-D-xylosidase belonging to family 39 glycoside hydrolase has been elucidated as a tetramer with each monomer being composed of three distinct regions: a catalytic domain of the canonical (β/α)8--TIM barrel fold, a β-sandwich domain and a small α-helical domain with the enzyme active site that binds to D-xylooligomers being present on the upper side of the barrel. Glycosylation is generally considered as one of the most important post-translational modifications of xylanases, but a few examples showed functional expression of eukaryotic xylanases in bacteria. The optimal ratio of these synergistic enzymes is very important in improving hydrolysis efficiency and reducing enzyme dosage but has hardly been addressed in literature. Xylanases have been used in traditional fields such as food, feed

  15. Mycelial glucoamylases produced by the thermophilic fungus Scytalidium thermophilum strains 15.1 and 15.8: purification and biochemical characterization Glucoamilases miceliais produzidas pelas linhagens 15.1 e 15.8 do fungo termofílico Scytalidium thermophilum: purificação e caracterização bioquímica

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    M.S. Ferreira-Nozawa

    2008-06-01

    Full Text Available Two strains (15.1 and 15.8 of the thermophilic fungus Scytalidium thermophilum produced high levels of intracellular glucoamylases, with potential for industrial applications. The isoform I of the glucoamylase produced by 15.1 strain was sequentially submitted to DEAE-Cellulose and CM-Cellulose chromatography, and purified 141-fold, with 5.45% recovery. The glucoamylase of strain 15.8 was purified 71-fold by CM-Cellulose and Concanavalin A-Sepharose chromatography, with 7.38% recovery. Temperature and pH optima were in the range of 50-60ºC and 5.0-6.0, respectively, using starch and maltose as substrates. The glucoamylase of S. thermophilum 15.8 was more stable (t50 > 60 min than that of S. thermophilum 15.1 (t50= 11-15 min, at 60ºC. The glucoamylase activities were enhanced by several ions (e.g. Mn2+ and Ca2+ and inhibited by β-mercaptoethanol. The glucoamylase from 15.1 strain showed a Km of 0.094 mg/ml and 0.029 mg/ml and Vmax of 202 U/mg prot and 109 U/mg prot, for starch and maltose, respectively. The hydrolysis products of starch and maltose, analyzed by TLC, demonstrated glucose as end product and confirming the character of the enzyme as glucoamylase. Differences were observed in relation to the products formed with maltose as substrate between the two strains studied. S. thermophilum 15.8 formed maltotriose in contrast with S. thermophilum 15.1.Duas linhagens (15.1 e 15.8 do fungo termofílico Scytalidium thermophilum se mostraram produtoras de grandes quantidades de glucoamilases, com potencial aplicação industrial. A isoforma I de glucoamilase produzida pela linhagem 15.1 foi submetida seqüencialmente a cromatografia em colunas de DEAE-celulose e CM-celulose, sendo purificada 141 vezes com porcentagem de recuperação de 5,45%. A glucoamilase da linhagem 15.8 foi purificada 71 vezes através do uso de colunas de cromatografia de CM-celulose e Concanavalina A-sepharose com porcentagem de recuperação de 7,38%. Temperatura e pH

  16. Accumulation of trehalose in the thermophilic fungus Chaetomium thermophilum var. coprophilum in response to heat or salt stress

    DEFF Research Database (Denmark)

    Jepsen, Helene Friborg; Jensen, B.

    2004-01-01

    The disaccharide trehalose, known to be an effective protectant against various kinds of stress, was observed to accumulate in the cytosol of Chaetomium thermophilum var. coprophilum during heat stress. Trehalose was apparently neither involved in the defence of C. thermophilum var. coprophilum...... against high concentrations of sodium chloride nor directly linked to thermophily. In C. thermophilum var. coprophilum three different trehalose hydrolyzing activities were eluted from a mono Q anion exchange column by sodium chloride concentrations of 0.10, 0.15 and 0.24 M, respectively....

  17. Simultaneous Silencing of Xylanase Genes in Botrytis cinerea

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    Néstor García

    2017-12-01

    Full Text Available The endo-β-1,4-xylanase BcXyn11A is one of several plant cell-wall degrading enzymes that the phytopathogenic fungus Botrytis cinerea secretes during interaction with its hosts. In addition to its enzymatic activity, this protein also acts as an elicitor of the defense response in plants and has been identified as a virulence factor. In the present work, other four endoxylanase coding genes (Bcxyn11B, Bcxyn11C, Bcxyn10A, and Bcxyn10B were identified in the B. cinerea genome and the expression of all five genes was analyzed by Q-RT- PCR in vitro and in planta. A cross-regulation between xylanase genes was identified analyzing their expression pattern in the ΔBcxyn11A mutant strain and a putative BcXyn11A-dependt induction of Bcxyn10B gene was found. In addition, multiple knockdown strains were obtained for the five endoxylanase genes by transformation of B. cinerea with a chimeric DNA construct composed of 50-nt sequences from the target genes. The silencing of each xylanase gene was analyzed in axenic cultures and during infection and the results showed that the efficiency of the multiple silencing depends on the growth conditions and on the cross-regulation between them. Although the simultaneous silencing of the five genes was observed by Q-RT-PCR when the silenced strains were grown on medium supplemented with tomato extract, the endoxylanase activity measured in the supernatants was reduced only by 40%. Unexpectedly, the silenced strains overexpressed the Bcxyn11A and Bcxyn11C genes during the infection of tomato leaves, making difficult the analysis of the role of the endo-β-1,4-xylanases in the virulence of the fungus.

  18. Cloning, sequencing, and sequence analysis of two novel plasmids from the thermophilic anaerobic bacterium Anaerocellum thermophilum

    DEFF Research Database (Denmark)

    Clausen, Anders; Mikkelsen, Marie Just; Schrøder, I.

    2004-01-01

    The nucleotide sequence of two novel plasmids isolated from the extreme thermophilic anaerobic bacterium Anaerocellum thermophilum DSM6725 (A. thermophilum), growing optimally at 70degreesC, has been determined. pBAS2 was found to be a 3653 bp plasmid with a GC content of 43%, and the sequence re...... with highest similarity to DNA repair protein from Campylobacter jejuni (25% aa). Orf34 showed similarity to sigma factors with highest similarity (28% aa) to the sporulation specific Sigma factor, Sigma 28(K) from Bacillus thuringiensis....

  19. Engineering better biomass-degrading ability into a GH11 xylanase using a directed evolution strategy

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    Song Letian

    2012-01-01

    Full Text Available Abstract Background Improving the hydrolytic performance of hemicellulases on lignocellulosic biomass is of considerable importance for second-generation biorefining. To address this problem, and also to gain greater understanding of structure-function relationships, especially related to xylanase action on complex biomass, we have implemented a combinatorial strategy to engineer the GH11 xylanase from Thermobacillus xylanilyticus (Tx-Xyn. Results Following in vitro enzyme evolution and screening on wheat straw, nine best-performing clones were identified, which display mutations at positions 3, 6, 27 and 111. All of these mutants showed increased hydrolytic activity on wheat straw, and solubilized arabinoxylans that were not modified by the parental enzyme. The most active mutants, S27T and Y111T, increased the solubilization of arabinoxylans from depleted wheat straw 2.3-fold and 2.1-fold, respectively, in comparison to the wild-type enzyme. In addition, five mutants, S27T, Y111H, Y111S, Y111T and S27T-Y111H increased total hemicellulose conversion of intact wheat straw from 16.7%tot. xyl (wild-type Tx-Xyn to 18.6% to 20.4%tot. xyl. Also, all five mutant enzymes exhibited a better ability to act in synergy with a cellulase cocktail (Accellerase 1500, thus procuring increases in overall wheat straw hydrolysis. Conclusions Analysis of the results allows us to hypothesize that the increased hydrolytic ability of the mutants is linked to (i improved ligand binding in a putative secondary binding site, (ii the diminution of surface hydrophobicity, and/or (iii the modification of thumb flexibility, induced by mutations at position 111. Nevertheless, the relatively modest improvements that were observed also underline the fact that enzyme engineering alone cannot overcome the limits imposed by the complex organization of the plant cell wall and the lignin barrier.

  20. Crystallization and preliminary X-ray analysis of a bifunctional catalase-phenol oxidase from Scytalidium thermophilum

    International Nuclear Information System (INIS)

    Sutay Kocabas, Didem; Pearson, Arwen R.; Phillips, Simon E. V.; Bakir, Ufuk; Ogel, Zumrut B.; McPherson, Michael J.; Trinh, Chi H.

    2009-01-01

    The bifunctional enzyme catalase-phenol oxidase from S. thermophilum was crystallized by the hanging-drop vapour-diffusion method in space group P2 1 and diffraction data were collected to 2.8 Å resolution. Catalase-phenol oxidase from Scytalidium thermophilum is a bifunctional enzyme: its major activity is the catalase-mediated decomposition of hydrogen peroxide, but it also catalyzes phenol oxidation. To understand the structural basis of this dual functionality, the enzyme, which has been shown to be a tetramer in solution, has been purified by anion-exchange and gel-filtration chromatography and has been crystallized using the hanging-drop vapour-diffusion technique. Streak-seeding was used to obtain larger crystals suitable for X-ray analysis. Diffraction data were collected to 2.8 Å resolution at the Daresbury Synchrotron Radiation Source. The crystals belonged to space group P2 1 and contained one tetramer per asymmetric unit

  1. A multipurpose immobilized biocatalyst with pectinase, xylanase and cellulase activities

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    Gupta Munishwar

    2007-06-01

    Full Text Available Abstract Background The use of immobilized enzymes for catalyzing various biotransformations is now a widely used approach. In recent years, cross-linked enzyme aggregates (CLEAs have emerged as a novel and versatile biocatalyst design. The present work deals with the preparation of a CLEA from a commercial preparation, Pectinex™ Ultra SP-L, which contains pectinase, xylanase and cellulase activities. The CLEA obtained could be used for any of the enzyme activities. The CLEA was characterized in terms of kinetic parameters, thermal stability and reusability in the context of all the three enzyme activities. Results Complete precipitation of the three enzyme activities was obtained with n-propanol. When resulting precipitates were subjected to cross-linking with 5 mM glutaraldehyde, the three activities initially present (pectinase, xylanase and cellulase were completely retained after cross-linking. The Vmax/Km values were increased from 11, 75 and 16 to 14, 80 and 19 in case of pectinase, xylanase and cellulase activities respectively. The thermal stability was studied at 50°C, 60°C and 70°C for pectinase, xylanase and cellulase respectively. Half-lives were improved from 17, 22 and 32 minutes to 180, 82 and 91 minutes for pectinase, xylanase and cellulase respectively. All three of the enzymes in CLEA could be reused three times without any loss of activity. Conclusion A single multipurpose biocatalyst has been designed which can be used for carrying out three different and independent reactions; 1 hydrolysis of pectin, 2 hydrolysis of xylan and 3 hydrolysis of cellulose. The preparation is more stable at higher temperatures as compared to the free enzymes.

  2. Xylanase production by a local fungal isolate, Aspergillus niger USM AI 1 via solid state

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    Ibrahim Che Omar

    2005-03-01

    Full Text Available Isolate USM A1 I which was identified to be Aspergillus niger was selected as a potential producer of xylanase via a solid state fermentation system (SSF using palm kernel cake (PKC as substrate. The modification of the physical conditions of the SSF system indicated that the xylanase activity was 23.97 U/g PKC at the moisture ratio of 1:0.75 of PKC: moistening agent with the inoculum size of 1¥104 spores/ml and cultivated at the ambient temperature (28±3ºC. The supplementation of additional carbon and nitrogen sources in the PKC medium could enhance enzyme productivity. The maximum production of xylanase and growth obtained with the supplementation of xylose at 0.75% (w/w were 25.40 U/g and 1.69 mg glucosamine/ g PKC. Moreover, the presence of NaNO3 at 0.075% (w/w as additional nitrogen source further enhanced xylanase production to 33.99 U/g PKC although the growth remained unchanged at about 1.67 mg glucosa- mine/g PKC. The optimized conditions showed an increased xylanase production by 157% compared to before the optimization of the SSF system. The xylanase productivity was 23.12 U/mg glucosamine after optimization and 11.72 U/mg glucosamine before optimization.

  3. The xylanase inhibitor TAXI-III counteracts the necrotic activity of a Fusarium graminearum xylanase in vitro and in durum wheat transgenic plants.

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    Moscetti, Ilaria; Faoro, Franco; Moro, Stefano; Sabbadin, Davide; Sella, Luca; Favaron, Francesco; D'Ovidio, Renato

    2015-08-01

    The xylanase inhibitor TAXI-III has been proven to delay Fusarium head blight (FHB) symptoms caused by Fusarium graminearum in transgenic durum wheat plants. To elucidate the molecular mechanism underlying the capacity of the TAXI-III transgenic plants to limit FHB symptoms, we treated wheat tissues with the xylanase FGSG_03624, hitherto shown to induce cell death and hydrogen peroxide accumulation. Experiments performed on lemmas of flowering wheat spikes and wheat cell suspension cultures demonstrated that pre-incubation of xylanase FGSG_03624 with TAXI-III significantly decreased cell death. Most interestingly, a reduced cell death relative to control non-transgenic plants was also obtained by treating, with the same xylanase, lemmas of TAXI-III transgenic plants. Molecular modelling studies predicted an interaction between the TAXI-III residue H395 and residues E122 and E214 belonging to the active site of xylanase FGSG_03624. These results provide, for the first time, clear indications in vitro and in planta that a xylanase inhibitor can prevent the necrotic activity of a xylanase, and suggest that the reduced FHB symptoms on transgenic TAXI-III plants may be a result not only of the direct inhibition of xylanase activity secreted by the pathogen, but also of the capacity of TAXI-III to avoid host cell death. © 2014 BSPP AND JOHN WILEY & SONS LTD.

  4. Cloning, expression, and characterization of a novel alkali-tolerant xylanase from alkaliphilic Bacillus sp. SN5.

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    Bai, Wenqin; Xue, Yanfen; Zhou, Cheng; Ma, Yanhe

    2015-01-01

    A xylanase gene (xyn11A) was cloned from the genomic library of alkalophilic Bacillus sp. SN5. It encoded a polypeptide of 366 amino acids, consisting of a family 11 glycoside hydrolase, a short linker region, and a family 36 carbohydrate-binding module (CBM). The intact xylanase Xyn11A and the CBM-linker-truncated Xyn11A-LC were expressed in Escherichia coli BL21 (DE3). Both purified recombinant proteins exhibited the highest activity at 55 °C. The optimal pH for Xyn11A activity was 7.5, whereas Xyn11A-LC showed a broad pH profile (>80% activity at pH 5.5-8.5) with optimal activity at pH 5.5 and 7.5-8.0. They had high alkali tolerance, retaining over 80% residual activity after preincubation at pH 8.5-11.0 at 37 °C for 1 H. Xyn11A-LC showed better thermal stability, lower affinity, and lower catalytic activity to insoluble xylan than Xyn11A, whereas its specific activity for soluble beechwood xylan (4,511.9 U/mg) was greater than that of Xyn11A (3,136.4 U/mg). These results implied that the CBM of Xyn11A could change the enzymatic properties and play a role in degrading insoluble xylan. Xyn11A-LC is a family 11 alkali-tolerant cellulase-free xylanase with high specific activity, which qualifies it as a potential candidate for industrial applications, especially in the paper industry. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

  5. Engineering Thermostable Microbial Xylanases Toward its Industrial Applications.

    Science.gov (United States)

    Kumar, Vishal; Dangi, Arun Kumar; Shukla, Pratyoosh

    2018-03-01

    Xylanases are one of the important hydrolytic enzymes which hydrolyze the β-1, 4 xylosidic linkage of the backbone of the xylan polymeric chain which consists of xylose subunits. Xylanases are mainly found in plant cell walls and are produced by several kinds of microorganisms such as fungi, bacteria, yeast, and some protozoans. The fungi are considered as most potent xylanase producers than that of yeast and bacteria. There is a broad series of industrial applications for the thermostable xylanase as an industrial enzyme. Thermostable xylanases have been used in a number of industries such as paper and pulp industry, biofuel industry, food and feed industry, textile industry, etc. The present review explores xylanase-substrate interactions using gene-editing tools toward the comprehension in improvement in industrial stability of xylanases. The various protein-engineering and metabolic-engineering methods have also been explored to improve operational stability of xylanase. Thermostable xylanases have also been used for improvement in animal feed nutritional value. Furthermore, they have been used directly in bakery and breweries, including a major use in paper and pulp industry as a biobleaching agent. This present review envisages some of such applications of thermostable xylanases for their bioengineering.

  6. C-Terminal carbohydrate-binding module 9_2 fused to the N-terminus of GH11 xylanase from Aspergillus niger.

    Science.gov (United States)

    Xu, Wenxuan; Liu, Yajuan; Ye, Yanxin; Liu, Meng; Han, Laichuang; Song, Andong; Liu, Liangwei

    2016-10-01

    The 9_2 carbohydrate-binding module (C2) locates natively at the C-terminus of the GH10 thermophilic xylanase from Thermotoga marimita. When fused to the C-terminus, C2 improved thermostability of a GH11 xylanase (Xyn) from Aspergillus niger. However, a question is whether the C-terminal C2 would have a thermostabilizing effect when fused to the N-terminus of a catalytic module. A chimeric enzyme, C2-Xyn, was created by step-extension PCR, cloned in pET21a(+), and expressed in E. coli BL21(DE3). The C2-Xyn exhibited a 2 °C higher optimal temperature, a 2.8-fold longer thermostability, and a 4.5-fold higher catalytic efficiency on beechwood xylan than the Xyn. The C2-Xyn exhibited a similar affinity for binding to beechwood xylan and a higher affinity for oat-spelt xylan than Xyn. C2 is a thermostabilizing carbohydrate-binding module and provides a model of fusion at an enzymatic terminus inconsistent with the modular natural terminal location.

  7. Structural characterization of the principal mRNA-export factor Mex67–Mtr2 from Chaetomium thermophilum

    Energy Technology Data Exchange (ETDEWEB)

    Aibara, Shintaro; Valkov, Eugene; Lamers, Meindert H. [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH (United Kingdom); Dimitrova, Lyudmila; Hurt, Ed [Biochemie-Zentrum der Universität Heidelberg, Im Neuenheimer Feld 328, 69120 Heidelberg (Germany); Stewart, Murray, E-mail: ms@mrc-lmb.cam.ac.uk [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH (United Kingdom)

    2015-06-27

    The crystal structures of the individual domains of the Mex67–Mtr2 complex from C. thermophilum have been determined and their arrangement in solution has been studied by SAXS. Members of the Mex67–Mtr2/NXF–NXT1 family are the principal mediators of the nuclear export of mRNA. Mex67/NXF1 has a modular structure based on four domains (RRM, LRR, NTF2-like and UBA) that are thought to be present across species, although the level of sequence conservation between organisms, especially in lower eukaryotes, is low. Here, the crystal structures of these domains from the thermophilic fungus Chaetomium thermophilum are presented together with small-angle X-ray scattering (SAXS) and in vitro RNA-binding data that indicate that, not withstanding the limited sequence conservation between different NXF family members, the molecules retain similar structural and RNA-binding properties. Moreover, the resolution of crystal structures obtained with the C. thermophilum domains was often higher than that obtained previously and, when combined with solution and biochemical studies, provided insight into the structural organization, self-association and RNA-binding properties of Mex67–Mtr2 that facilitate mRNA nuclear export.

  8. Structural characterization of the principal mRNA-export factor Mex67–Mtr2 from Chaetomium thermophilum

    International Nuclear Information System (INIS)

    Aibara, Shintaro; Valkov, Eugene; Lamers, Meindert H.; Dimitrova, Lyudmila; Hurt, Ed; Stewart, Murray

    2015-01-01

    The crystal structures of the individual domains of the Mex67–Mtr2 complex from C. thermophilum have been determined and their arrangement in solution has been studied by SAXS. Members of the Mex67–Mtr2/NXF–NXT1 family are the principal mediators of the nuclear export of mRNA. Mex67/NXF1 has a modular structure based on four domains (RRM, LRR, NTF2-like and UBA) that are thought to be present across species, although the level of sequence conservation between organisms, especially in lower eukaryotes, is low. Here, the crystal structures of these domains from the thermophilic fungus Chaetomium thermophilum are presented together with small-angle X-ray scattering (SAXS) and in vitro RNA-binding data that indicate that, not withstanding the limited sequence conservation between different NXF family members, the molecules retain similar structural and RNA-binding properties. Moreover, the resolution of crystal structures obtained with the C. thermophilum domains was often higher than that obtained previously and, when combined with solution and biochemical studies, provided insight into the structural organization, self-association and RNA-binding properties of Mex67–Mtr2 that facilitate mRNA nuclear export

  9. Extractive fermentation of xylanase from Aspergillus tamarii URM 4634 in a bioreactor.

    Science.gov (United States)

    da Silva, Anna Carolina; Soares de França Queiroz, Alana Emília; Evaristo dos Santos Nascimento, Talita Camila; Rodrigues, Cristine; Gomes, José Erick Galindo; Souza-Motta, Cristina Maria; Porto de Souza Vandenberghe, Luciana; Valente de Medeiros, Erika; Moreira, Keila Aparecida; Herculano, Polyanna Nunes

    2014-08-01

    Of the many reported applications for xylanase, its use as a food supplement has played an important role for monogastric animals, because it can improve the utilisation of nutrients. The aim of this work was to produce xylanase by extractive fermentation in an aqueous two-phase system using Aspergillus tamarii URM 4634, increasing the scale of production in a bioreactor, partially characterising the xylanase and evaluating its influence on monogastric digestion in vitro. Through extractive fermentation in a bioreactor, xylanase was obtained with an activity of 331.4 U mL(-1) and 72% yield. The xylanase was stable under variable pH and temperature conditions, and it was optimally active at pH 3.6 and 90 °C. Xylanase activity potentiated the simulation of complete monogastric digestion by 6%, and only Mg2+ inhibited its activity. This process provides a system for efficient xylanase production by A. tamarii URM 4634 that has great potential for industrial use.

  10. Cloning of a novel xylanase gene from a newly isolated Fusarium sp. Q7-31 and its expression in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Zhan-Ling Xie

    2012-03-01

    Full Text Available A strain of Q7-31 was isolated from Qinghai-Tibet Plateau and was identified as Fusarium sp. based on its morphological characteristics and ITS rDNA gene sequence analysis. It has the highest capacity of degrading cell wall activity compared with other 11 strains. To do research on its xylanase activity of Fusarium sp. Q7-31 while the degrading the rice cell walls, the complete gene xyn8 that encodes endo-1, 4-β-xylanase secreted by Fusarium sp. Q7-31 was cloned and sequenced. The coding region of the gene is separated by two introns of 56bp and 55bp. It encodes 230 amino acid residues of a protein with a calculated molecular weight of 25.7 kDa. The animo acids sequence of xyn8 gene has higher similarity with those of family 11 of glycosyl hydrolases reported from other microorganisms. The nature peptide encodeing cDNA was subcloned into pGEX5x-1 expression vector. The recombinant plasmid was expressed in Escherichia coli BL21-CodonPlus (DE3-RIL, and xylanase activity was measured. The expression fusion protein was identified by SDS-PAGE and Western blotting, a new specific band of about 52kDa was identified when induced by IPTG. Enzyme activity assay verified the recombinants proteins as a xylanase. A maxium activity of 2.34U/ mg, the xylanase had optimal activity at pH 6.0 and temperature 40ºC .

  11. Gene cloning, overexpression, and characterization of a xylanase from Penicillium sp. CGMCC 1669.

    Science.gov (United States)

    Liu, Wanli; Shi, Pengjun; Chen, Qiang; Yang, Peilong; Wang, Guozeng; Wang, Yaru; Luo, Huiying; Yao, Bin

    2010-09-01

    A xylanase-encoding gene, xyn11F63, was isolated from Penicillium sp. F63 CGMCC1669 using degenerated polymerase chain reaction (PCR) and thermal asymmetric interlaced (TAIL)-PCR techniques. The full-length chromosomal gene consists of 724 bp, including a 73-bp intron, and encodes a 217 amino acid polypeptide. The deduced amino acid sequence of xyn11F63 shows the highest identity of 70% to the xylanase from Penicillium sp. strain 40, which belongs to glycosyl hydrolases family 11. The gene was overexpressed in Pichia pastoris, and its activity in the culture medium reached 516 U ml(-1). After purification to electrophoretic homogeneity, the enzyme showed maximal activity at pH 4.5 and 40 degrees C, was stable at acidic buffers of pH 4.5-9.0, and was resistant to proteases (proteinase K, trypsin, subtilisin A, and alpha-chymotrypsin). The specific activity, K (m), and V (max) for oat spelt xylan substrate was 7,988 U mg(-1), 22.2 mg ml(-1), and 15,105.7 micromol min(-1) mg(-1), respectively. These properties make XYN11F63 a potential economical candidate for use in feed and food industrial applications.

  12. Xylanase production by a newly isolated Aspergillus niger SS7 in submerged culture.

    Science.gov (United States)

    Bakri, Yasser; Al-Jazairi, Manal; Al-Kayat, Ghassan

    2008-01-01

    Xylanase production by a newly isolated Aspergillus niger SS7 was studied in submerged culture. The optimum initial pH for xylanase production was found to be 7.0. Different agricultural and industrial wastes were evaluated for their ability to induce xylanase production by this isolate. The best xylanase production (293.82 IU/ml) was recorded at 3% (w/v) corn cob hulls after 120 h of incubation. The Aspergillus niger SS7 isolate grown in a simple medium, proved to be a promising microorganism for xylanase production.

  13. Xylanase production by Trichoderma harzianum E58

    Energy Technology Data Exchange (ETDEWEB)

    Senior, D.J.; Mayers, P.R.; Saddler, J.N. (Fortintek Canada Corp., Ottawa, ON (Canada). Dept. of Biotechnology and Chemistry)

    1989-12-01

    Growth of Trichoderma harzianum E58 on hemicellulose-rich media, both in batch and fermentor cultures, resulted in independent profiles of the production of xylanase and endoglucanase enzymes. Dramatic differences in the ratio of xylanase to endoglucanase activities were observed among cultures grown on cellulose-rich Solka Floc and xylan. These results indicated that the induction of xylanases and cellulases was likely to be under separate regulatory control. The specific activity and amount of xylanases produced were found to be dependent on the concentration of xylan in the growth media. Growth on oat spelts xylan or the hemicellulose-rich, watersoluble fraction from steam-treated aspenwood (SEA-WS) greatly enhanced the production of xylanases and xylosidase in the culture filtrates. Constitutive levels of xylanase and endoglucanase enzymes were detected during growth of the fungus on glucose. (orig.).

  14. Xylanases of thermophilic bacteria from Icelandic hot springs

    Energy Technology Data Exchange (ETDEWEB)

    Pertulla, M; Raettoe, M; Viikari, L [VTT, Biotechnical Lab., Espoo (Finland); Kondradsdottir, M [Dept. of Biotechnology, Technological Inst. of Iceland, Reykjavik (Iceland); Kristjansson, J K [Dept. of Biotechnology, Technological Inst. of Iceland, Reykjavik (Iceland) Inst. of Biotechnology, Iceland Univ., Reykjavik (Iceland)

    1993-02-01

    Thermophilic, aerobic bacteria isolated from Icelandic hot springs were screened for xylanase activity. Of 97 strains tested, 14 were found to be xylanase positive. Xylanase activities up to 12 nkat/ml were produced by these strains in shake flasks on xylan medium. The xylanases of the two strains producing the highest activities (ITI 36 and ITI 283) were similar with respect to temperature and pH optima (80deg C and pH 8.0). Xylanase production of strain ITI 36 was found to be induced by xylan and xylose. Xylanase activity of 24 nkat/ml was obtained with this strain in a laboratory-scale-fermentor cultivation on xylose medium. [beta]-Xylosidase activity was also detected in the culture filtrate. The thermal half-life of ITI 36 xylanase was 24 h at 70deg C. The highest production of sugars from hydrolysis of beech xylan was obtained at 70deg C, although xylan depolymerization was detected even up to 90deg C. (orig.).

  15. Molecular characterization of a Xylanase-producing fungus isolated from fouled soil

    Directory of Open Access Journals (Sweden)

    Punniavan Sakthiselvan

    2014-12-01

    Full Text Available Xylanase (EC 3. 2. 1. 8, hydrolyzes xylo-oligosaccharides into D-xylose and required for complete hydrolysis of native cellulose and biomass conversion. It has broad range of applications in the pulp and paper, pharmaceutical and Agri-food industries. Fifty fungal species were isolated from the fouled soil around an oil refinery and screened for the production of xylanase enzyme by enrichment culture techniques. The isolated fungal strain was identified as Hypocrea lixii SS1 based on the results of biochemical tests and 18s rRNA sequencing. The phylogenetic tree was constructed using the MEGA 5 software. Further, Hypocrea lixii SS1 was tested for the ability to utilize the sunflower oil sludge (waste from the oil industry as the sole carbon source for xylanase production. The growth characteristics of Hypocrea lixii SS1 were also studied and maximum growth was found on the 7th day of incubation. The fungus showed a remarkable xylanase production of 38.9 U/mL. Xylanase was purified using a combination of 0-50% NH4SO2 precipitation, DEAE-sepharose and Sephacryl S-200 chromatography. Single peak obtained in RP-HPLC confirms the purity of xylanase. Further the enzyme produced was affirmed as xylanase with its molecular weight (29 kDa using SDS-PAGE.

  16. Thermophilic xylanases: from bench to bottle.

    Science.gov (United States)

    Basit, Abdul; Liu, Junquan; Rahim, Kashif; Jiang, Wei; Lou, Huiqiang

    2018-01-17

    Lignocellulosic biomass is a valuable raw material. As technology has evolved, industrial interest in new ways to take advantage of this raw material has grown. Biomass is treated with different microbial cells or enzymes under ideal industrial conditions to produce the desired products. Xylanases are the key enzymes that degrade the xylosidic linkages in the xylan backbone of the biomass, and commercial enzymes are categorized into different glycoside hydrolase families. Thermophilic microorganisms are excellent sources of industrially relevant thermostable enzymes that can withstand the harsh conditions of industrial processing. Thermostable xylanases display high-specific activity at elevated temperatures and distinguish themselves in biochemical properties, structures, and modes of action from their mesophilic counterparts. Natural xylanases can be further improved through genetic engineering. Rapid progress with genome editing, writing, and synthetic biological techniques have provided unlimited potential to produce thermophilic xylanases in their natural hosts or cell factories including bacteria, yeasts, and filamentous fungi. This review will discuss the biotechnological potential of xylanases from thermophilic microorganisms and the ways they are being optimized and produced for various industrial applications.

  17. Growth characteristics of the thermophilic fungus Scytalidium thermophilum in relation to production of mushroom compost.

    NARCIS (Netherlands)

    Wiegant, W.M.

    1992-01-01

    Scytalidium thermophilum is an important thermophilic fungus in the production of mushroom compost. I investigated the characteristics of this organism and present a simple model with which fungal growth in compost can be described. The model is used to predict better circumstances for rapid indoor

  18. Xylanases and Their Applications in Baking Industry

    Directory of Open Access Journals (Sweden)

    Masood Sadiq Butt

    2008-01-01

    Full Text Available Xylan is the second most abundant polysaccharide and a major component of plant cell wall. Cereal xylans contain large quantities of L-arabinose and are therefore, often referred to as arabinoxylans. Xylanases are hydrolytic enzymes, which randomly cleave the β-1,4 backbone of this complex plant cell wall polysaccharide. Different species of Aspergillus and Trichoderma produce these enzymes. Xylanases are of great value in baking as they have been found to improve the bread volume, crumb structure and reduce stickiness. When xylanases are used at optimum levels, they play a significant role in increasing shelf life of bread and reduce bread staling. There is an increasing trend in baking industry towards the application of xylanases in bread production. This review discusses the application of xylanase in the bakery industry, alone and in combination with other enzymes when it shows synergism in the action with them.

  19. Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanase

    Directory of Open Access Journals (Sweden)

    Liu Liangwei

    2012-06-01

    Full Text Available Abstract Background Domain fusion is limited at enzyme one terminus. The issue was explored by swapping a mesophilic Aspergillus niger GH11 xylanase (Xyn with a hyper-thermophilic Thermotoga maritima glucanase (Glu to construct two chimeras, Xyn-Glu and Glu-Xyn, with an intention to create thermostable xylanase containing glucanase activity. Results When expressed in E. coli BL21(DE3, the two chimeras exhibited bi-functional activities of xylanase and glucanase. The Xyn-Glu Xyn moiety had optimal reaction temperature (Topt at 50 °C and thermal in-activation half-life (t1/2 at 50 °C for 47.6 min, compared to 47 °C and 17.6 min for the Xyn. The Glu-Xyn Xyn moiety had equivalent Topt to and shorter t1/2 (5.2 min than the Xyn. Both chimera Glu moieties were more thermostable than the Glu, and the three enzyme Topt values were higher than 96 °C. The Glu-Xyn Glu moiety optimal pH was 5.8, compared to 3.8 for the Xyn-Glu Glu moiety and the Glu. Both chimera two moieties cooperated with each other in degrading substrates. Conclusions Domain-swapping created different effects on each moiety properties. Fusing the Glu domain at C-terminus increased the xylanase thermostability, but fusing the Glu domain at N-terminus decreased the xylanase thermostability. Fusing the Xyn domain at either terminus increased the glucanase thermostability, and fusing the Xyn domain at C-terminus shifted the glucanase pH property 2 units higher towards alkaline environments. Fusing a domain at C-terminus contributes more to enzyme catalytic activity; whereas, fusing a bigger domain at N-terminus disturbs enzyme substrate binding affinity.

  20. Production and characterization of xylanase from Thielaviopsis basicola

    Energy Technology Data Exchange (ETDEWEB)

    Ghosh, V K; Deb, J K

    1988-08-01

    The black rot fungus Thielaviopsis basicola has the ability to grow on cellulosic biomass, producing xylanase. Of the four cellulosic substrates tested, rice straw was found to be the best for production of xylanase. A xylanase activity of 34 U/ml was obtained with rice straw which was more than three times that obtained with larchwood xylan. The ..beta..-xylosidase activities obtained with these two substrates were 0.05 U/ml and 0.016 U/ml respectively. Both enzymes are active at pH 5 but the temperature optima of xylanase and ..beta..-xylosidase activities are 60/sup 0/C and 40/sup 0/C respectively. The xylanase activity is stable over a pH range of 4-8 but the stability towards temperature falls sharply above 50/sup 0/C.

  1. A comparison of two xylanases from the thermophilic fungi Thielavia terrestris and Thermoascus crustaceus

    Energy Technology Data Exchange (ETDEWEB)

    Gilbert, M [Ottawa Univ., Dept. of Biology, ON (Canada); Yaguchi, M [Inst. for Biological Sciences, National Research Council of Canada, Ottawa, ON (Canada); Watson, D C [Inst. for Biological Sciences, National Research Council of Canada, Ottawa, ON (Canada); Wong, K K.Y. [Chair of Forest Products Biotechnology, Faculty of Forestry, British Columbia Univ., Vancouver, BC (Canada); Breuil, C [Chair of Forest Products Biotechnology, Faculty of Forestry, British Columbia Univ., Vancouver, BC (Canada); Saddler, J N [Chair of Forest Products Biotechnology, Faculty of Forestry, British Columbia Univ., Vancouver, BC (Canada)

    1993-12-01

    Two thermophilic xylanases (xylanase II from Thielavia terrestris 255B and the 32-kDa xylanase from Thermoascus crustaceus 235E) were studied to determine if they had different and complementary modes of action when they hydrolysed various types of xylans. Partial amino acid sequencing showed that these two enzymes belonged to different families of [beta]-1,4-glycanases. Xylanase II achieved faster solubilization of insoluble xylan whereas the 32-kDa xylanase was more effective in producing xylose and short xylooligomers. An assessment of the combined hydrolytic action of the two xylanases did not reveal any co-operative action. The sugars released when the two thermophilic xylanases were used together were almost identical to those released when the 32-kDa xylanase acted alone. The two xyalanses were able to remove about 12% of the xylan remaining in an aspen kraft pulp. This indicated that either one of these thermophilic enzymes may be useful for enhancing the bleaching of kraft pulps. (orig.)

  2. A xylanase-aided enzymatic pretreatment facilitates cellulose nanofibrillation.

    Science.gov (United States)

    Long, Lingfeng; Tian, Dong; Hu, Jinguang; Wang, Fei; Saddler, Jack

    2017-11-01

    Although biological pretreatment of cellulosic fiber based on endoglucanases has shown some promise to facilitate cellulose nanofibrillation, its efficacy is still limited. In this study, a xylanase-aided endoglucanase pretreatment was assessed on the bleached hardwood and softwood Kraft pulps to facilitate the downstream cellulose nanofibrillation. Four commercial xylanase preparations were compared and the changes of major fiber physicochemical characteristics such as cellulose/hemicellulose content, gross fiber properties, fiber morphologies, cellulose accessibility/degree of polymerization (DP)/crystallinity were systematically evaluated before and after enzymatic pretreatment. It showed that the synergistic cooperation between endoglucanase and certain xylanase (Biobrite) could efficiently "open up" the hardwood Kraft pulp with limited carbohydrates degradation (cellulose nanofibrillation during mild sonication process (90Wh) with more uniform disintegrated nanofibril products (50-150nm, as assessed by scanning electron microscopy and UV-vis spectroscopy). Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Characterization of Xylanase Streptomyces spp. SKK1-8

    Directory of Open Access Journals (Sweden)

    ANJA MERYANDINI

    2006-12-01

    Full Text Available Streptomyces spp. SKK1-8 producing xylanase was isolated from soil sample from Sukabumi West Java. The xylanase have an optimum condition at pH 6 and 50 °C. Addition of 5 mM Cu2+ decreased the xylanase activity up to about 77%, whereas not by other cations. The xylanase was stable at 3 °C for 48 hours, and the enzyme half lifetime was 1 hour 45 minute at 50 °C. This xylanase showed the highest activity on oatspelt xylan, and their molecular masses were estimated approximately 16.80, 15.21, and 13.86 kDa. HPLC analysis showed that xylosa and arabinosa were the main hydrolytic product of birchwood xylan.

  4. Analysis of functional xylanases in xylan degradation by Aspergillus niger E-1 and characterization of the GH family 10 xylanase XynVII.

    Science.gov (United States)

    Takahashi, Yui; Kawabata, Hiroaki; Murakami, Shuichiro

    2013-01-01

    Xylanases produced by Aspergillus niger are industrially important and many types of xylanases have been reported. Individual xylanases have been well studied for their enzymatic properties, gene cloning, and heterologous expression. However, less attention has been paid to the relationship between xylanase genes carried on the A. niger genome and xylanases produced by A. niger strains. Therefore, we examined xylanase genes encoded on the genome of A. niger E-1 and xylanases produced in culture. Seven putative xylanase genes, xynI-VII (named in ascending order of the molecular masses of the deduced amino acid sequences), were amplified from the strain E-1 genome using primers designed from the genome sequence of A. niger CBS 513.88 by PCR and phylogenetically classified into three clusters. Additionally, culture supernatant analysis by DE52 anion-exchange column chromatography revealed that this strain produced three xylanases, XynII, XynIII, and XynVII, which were identified by N-terminal amino acid sequencing and MALDI-TOF-MS analyses, in culture when gown in 0.5% xylan medium supplemented with 50 mM succinate. Furthermore, XynVII, the only GH family 10 xylanase in A. niger E-1, was purified and characterized. The purified enzyme showed a single band with a molecular mass of 35 kDa by SDS-PAGE. The highest activity of purified XynVII was observed at 55°C and pH 5.5. The enzyme was stable in the broad pH range of 3-10 and up to 60°C and was resistant to most metal ions and modifying regents. XynVII showed high specificity against beechwood xylan with K m and V max values of 2.8 mg mL(-1) and 127 μmol min(-1)mg(-1), respectively. TLC and MALDI-TOF-MS analyses showed that the final hydrolyzed products of the enzyme from beechwood xylan were xylose, xylobiose, and xylotriose substituted with a 4-o-metylglucuronic acid residue.

  5. Improving the temperature characteristics and catalytic efficiency of a mesophilic xylanase from Aspergillus oryzae, AoXyn11A, by iterative mutagenesis based on in silico design.

    Science.gov (United States)

    Li, Xue-Qing; Wu, Qin; Hu, Die; Wang, Rui; Liu, Yan; Wu, Min-Chen; Li, Jian-Fang

    2017-12-01

    To improve the temperature characteristics and catalytic efficiency of a glycoside hydrolase family (GHF) 11 xylanase from Aspergillus oryzae (AoXyn11A), its variants were predicted based on in silico design. Firstly, Gly 21 with the maximum B-factor value, which was confirmed by molecular dynamics (MD) simulation on the three-dimensional structure of AoXyn11A, was subjected to site-saturation mutagenesis. Thus, one variant with the highest thermostability, AoXyn11A G21I , was selected from the mutagenesis library, E. coli/Aoxyn11A G21X (X: any one of 20 amino acids). Secondly, based on the primary structure multiple alignment of AoXyn11A with seven thermophilic GHF11 xylanases, AoXyn11A Y13F or AoXyn11A G21I-Y13F , was designed by replacing Tyr 13 in AoXyn11A or AoXyn11A G21I with Phe. Finally, three variant-encoding genes, Aoxyn11A G21I , Aoxyn11A Y13F and Aoxyn11A G21I-Y13F , were constructed by two-stage whole-plasmid PCR method, and expressed in Pichia pastoris GS115, respectively. The temperature optimum (T opt ) of recombinant (re) AoXyn11A G21I-Y13F was 60 °C, being 5 °C higher than that of reAoXyn11A G21I or reAoXyn11A Y13F , and 10 °C higher than that of reAoXyn11A. The thermal inactivation half-life (t 1/2 ) of reAoXyn11A G21I-Y13F at 50 °C was 240 min, being 40-, 3.4- and 2.5-fold longer than those of reAoXyn11A, reAoXyn11A G21I and reAoXyn11A Y13F . The melting temperature (T m ) values of reAoXyn11A, reAoXyn11A G21I , reAoXyn11A Y13F and reAoXyn11A G21I-Y13F were 52.3, 56.5, 58.6 and 61.3 °C, respectively. These findings indicated that the iterative mutagenesis of both Gly21Ile and Tyr13Phe improved the temperature characteristics of AoXyn11A in a synergistic mode. Besides those, the catalytic efficiency (k cat /K m ) of reAoXyn11A G21I-Y13F was 473.1 mL mg -1 s -1 , which was 1.65-fold higher than that of reAoXyn11A.

  6. A thermostable Gloeophyllum trabeum xylanase with potential for the brewing industry.

    Science.gov (United States)

    Wang, Xiaoyu; Luo, Huiying; Yu, Wangning; Ma, Rui; You, Shuai; Liu, Weina; Hou, Lingyu; Zheng, Fei; Xie, Xiangming; Yao, Bin

    2016-05-15

    A xylanase gene of glycoside hydrolase family 10, GtXyn10, was cloned from Gloeophyllum trabeum CBS 900.73 and expressed in Pichia pastoris GS115. Purified recombinant GtXyn10 exhibited significant activities to xylan (100.0%), lichenan (11.2%), glucan (15.2%) and p-nitrophenol-β-cellobiose (18.6%), demonstrated the maximum xylanase and glucanase activities at pH 4.5-5.0 and 75°C, retained stability over the pH range of 2.0-7.5 and at 70°C, and was resistant to pepsin and trypsin, most metal ions and SDS. Multiple sequence alignment and modeled-structure analysis identified a unique Gly48 in GtXyn10, and site-directed mutagenesis of Gly48 to Lys improved the temperature optimum up to 80°C. Under simulated mashing conditions, GtXyn10 (80U) reduced the mash viscosity by 12.8% and improved the filtration rate by 31.3%. All these properties above make GtXyn10 attractive for potential applications in the feed and brewing industries. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Cloning, sequencing and expression of a novel xylanase cDNA from ...

    African Journals Online (AJOL)

    A strain SH 2016, capable of producing xylanase, was isolated and identified as Aspergillus awamori, based on its physiological and biochemical characteristics as well as its ITS rDNA gene sequence analysis. A xylanase gene of 591 bp was cloned from this newly isolated A. awamori and the ORF sequence predicted a ...

  8. Highly thermostable xylanase production from a thermophilic Geobacillus sp. strain WSUCF1 utilizing lignocellulosic biomass

    Directory of Open Access Journals (Sweden)

    Aditya eBhalla

    2015-06-01

    Full Text Available AbstractEfficient enzymatic hydrolysis of lignocellulose to fermentable sugars requires a complete repertoire of biomass deconstruction enzymes. Hemicellulases play an important role in hydrolyzing hemicellulose component of lignocellulose to xylo-oligosaccharides and xylose. Thermostable xylanases have been a focus of attention as industrially important enzymes due to their long shelf life at high temperatures. Geobacillus sp. strain WSUCF1 produced thermostable xylanase activity (crude xylanase cocktail when grown on xylan or various inexpensive untreated and pretreated lignocellulosic biomasses such as prairie cord grass and corn stover. The optimum pH and temperature for the crude xylanase cocktail were 6.5 and 70ºC, respectively. The WSUCF1 crude xylanase was found to be highly thermostable with half-lives of 18 and 12 days at 60 and 70ºC, respectively. At 70ºC, rates of xylan hydrolysis were also found to be better with the WSUCF1 secretome than those with commercial enzymes, i.e., for WSUCF1 crude xylanase, CellicHTec2, and AccelleraseXY, the percent xylan conversions were 68.9, 49.4, and 28.92, respectively. To the best of our knowledge, WSUCF1 crude xylanase cocktail is among the most thermostable xylanases produced by thermophilic Geobacillus spp. and other thermophilic microbes (optimum growth temperature ≤70ºC. High thermostability, activity over wide range of temperatures, and better xylan hydrolysis than commercial enzymes make WSUCF1 crude xylanase suitable for thermophilic lignocellulose bioconversion processes.

  9. Comparison of kinetic characteristics of xylanases from Aspergillus ...

    African Journals Online (AJOL)

    LAB

    2013-05-08

    May 8, 2013 ... convert the substrate into products. The xylanase from A. ... be evidence that this enzyme is less active than the xylanase from A. niger. Moreover, the ..... lignocellulosic biomass structural polysaccharides. Biotechnol. Agron.

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

    African Journals Online (AJOL)

    MIET

    2013-05-15

    May 15, 2013 ... processing (Collins et al., 2005). Frequent ... xylanases (Stricker et al., 2008). The use of cheaper lignocellulosic residues viz. wheat bran, wheat straw, corn cob and sugarcane bagasse can be used as growth ..... Table 3. Effect of different temperature on xylanase activity from A. fumigatus (enzyme reaction.

  11. Production of Sporotrichum thermophile xylanase by solid state fermentation utilizing deoiled Jatropha curcas seed cake and its application in xylooligosachharide synthesis.

    Science.gov (United States)

    Sadaf, Ayesha; Khare, S K

    2014-02-01

    De-oiled Jatropha curcas seed cake, a plentiful by-product of biodiesel industry was used as substrate for the production of a useful xylanase from Sporotrichum thermophile in solid state fermentation. Under the optimized conditions, 1025U xylanase/g (deoiled seed cake) was produced. The xylanase exhibited half life of 4h at 45°C and 71.44min at 50°C respectively. It was stable in a broad pH range of 7.0-11.0. Km and Vmax were 12.54mg/ml and 454.5U/ml/min respectively. S. thermophile xylanase is an endoxylanase free of exoxylanase activity, hence advantageous for xylan hydrolysis to produce xylooligosachharides. Hydrolysis of oat spelt xylan by S. thermophile xylanase yielded 73% xylotetraose, 15.4% xylotriose and 10% xylobiose. The S. thermophile endoxylanase thus seem potentially useful in the food industries. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Improvement of xylanase production by a parasexual cross between Aspergillus niger strains

    Directory of Open Access Journals (Sweden)

    Octavio Loera

    2003-03-01

    Full Text Available A diploid strain (D4 isolated via parasexual recombination between two Aspergillus niger xylanase overproducing mutants was characterised in terms of enzyme production and catabolite repression by glucose. This strain increased xylanase production (607 nkat/ml, which was nearly 100% higher than titers achieved by the wild type strain (305 nkat/ml and 28% higher than the best mutant used to induce parasexual cycle. Diploid D4 was also less sensitive to carbon catabolite repression by glucose, since xylanolytic activity was detected under conditions normally repressing production by the wild type strain. No decrease in maximal xylanase levels was observed in the presence of glucose for diploid D4.Um cepa diplóide (D4 isolada por combinação parasexual entre dois Aspergillus niger, mutantes superprodutores de xylanase foi caracterizado através da produção de (607 nkat/ml e repressão catabólica por glicose. Essa cepa aumenta a produção de xylanase em mais de 100% em comparação com uma cepa selvagem (305 nkat/ml e 28% superior do que o melhor mutante usado para induzir o ciclo parasexual. A cepa diplóide D4 foi também menos sensível a repressão catabólica pela glicose, sendo que a atividade xylanolitica foi detectada sob condições normalmente de produção repressiva pela cepa selvagem. Não foi observado um decréscimo na produção máxima de xylanase em presença de glicose para o diplóide D4.

  13. A novel cellulase free alkaliphilic xylanase from alkali tolerant Penicillium citrinum: production, purification and characterization.

    Science.gov (United States)

    Dutta, T; Sengupta, R; Sahoo, R; Sinha Ray, S; Bhattacharjee, A; Ghosh, S

    2007-02-01

    The enzymatic hydrolysis of xylan has potential economic and environment-friendly applications. Therefore, attention is focused here on the discovery of new extremophilic xylanase in order to meet the requirements of industry. An extracellular xylanase was purified from the culture filtrate of P. citrinum grown on wheat bran bed in solid substrate fermentation. Single step purification was achieved using hydrophobic interaction chromatography. The purified enzyme showed a single band on SDS-PAGE with an apparent molecular weight of c. 25 kDa and pI of 3.6. Stimulation of the activity by beta mercaptoethanol, dithiotheritol (DTT) and cysteine was observed. Moderately thermostable xylanase showed optimum activity at 50 degrees C at pH 8.5. Xylanase purified from P. citrinum was alkaliphilic and moderately thermostable in nature. The present work reports for the first time the purification and characterization of a novel endoglucanase free alkaliphilic xylanase from the alkali tolerant fungus Penicillium citrinum. The alkaliphilicity and moderate thermostability of this xylanase may have potential implications in paper and pulp industries.

  14. Purification and Characterization of Streptomyces sp. SKK1-8 xylanase

    Directory of Open Access Journals (Sweden)

    Nunuk Widhyastuti

    2008-11-01

    Full Text Available Streptomyces sp. SKK1-8 is a xylanase produced bacteria. Purified xylanase has an optimum condition at pH 4.5 and 50oC. The molecular mass of purified xylanase were determined to be 14.4 kDa and 13.4 kDa. The xylanase was capable of hydrolysing p-NP-α-L-arabinofuranoside, p-NP-β-D-xylanopiranoside, p-NP-β-D-glucopiranoside, p-NP-α-D-galactopiranoside. The Km and Vmax values at 50oC measured on Birchwood xylan were 0.101 mg/ml and 0.1796 μmoles/minute/ml.

  15. Screening and production study of microbial xylanase producers from Brazilian Cerrado.

    Science.gov (United States)

    Alves-Prado, Heloiza Ferreira; Pavezzi, Fabiana Carina; Leite, Rodrigo Simões Ribeiro; de Oliveira, Valéria Maia; Sette, Lara Durães; Dasilva, Roberto

    2010-05-01

    Hemicelluloses are polysaccharides of low molecular weight containing 100 to 200 glycosidic residues. In plants, the xylans or the hemicelluloses are situated between the lignin and the collection of cellulose fibers underneath. The xylan is the most common hemicellulosic polysaccharide in cell walls of land plants, comprising a backbone of xylose residues linked by beta-1,4-glycosidic bonds. So, xylanolytic enzymes from microorganism have attracted a great deal of attention in the last decade, particularly because of their biotechnological characteristics in various industrial processes, related to food, feed, ethanol, pulp, and paper industries. A microbial screening of xylanase producer was carried out in Brazilian Cerrado area in Selviria city, Mato Grosso do Sul State, Brazil. About 50 bacterial strains and 15 fungal strains were isolated from soil sample at 35 degrees C. Between these isolated microorganisms, a bacterium Lysinibacillus sp. and a fungus Neosartorya spinosa as good xylanase producers were identified. Based on identification processes, Lysinibacillus sp. is a new species and the xylanase production by this bacterial genus was not reported yet. Similarly, it has not reported about xylanase production from N. spinosa. The bacterial strain P5B1 identified as Lysinibacillus sp. was cultivated on submerged fermentation using as substrate xylan, wheat bran, corn straw, corncob, and sugar cane bagasse. Corn straw and wheat bran show a good xylanase activity after 72 h of fermentation. A fungus identified as N. spinosa (strain P2D16) was cultivated on solid-state fermentation using as substrate source wheat bran, wheat bran plus sawdust, corn straw, corncob, cassava bran, and sugar cane bagasse. Wheat bran and corncobs show the better xylanase production after 72 h of fermentation. Both crude xylanases were characterized and a bacterial xylanase shows optimum pH for enzyme activity at 6.0, whereas a fungal xylanase has optimum pH at 5.0-5.5. They were

  16. Purification and characterization of a thermostable hypothetical xylanase from Aspergillus oryzae HML366.

    Science.gov (United States)

    He, Haiyan; Qin, Yongling; Li, Nan; Chen, Guiguang; Liang, Zhiqun

    2015-03-01

    In the current study, fermentation broth of Aspergillus oryzae HML366 in sugar cane bagasse was subjected to ultrafiltration and ion exchange chromatography, and two xylanases, XynH1 and XynH2, were purified. Time-of-flight mass spectrometry coupled with SDS-PAGE analysis revealed that XynH1 is identical to the hypothetical A. oryzae RIB40 protein XP_001826985.1, with a molecular weight of 33.671 kDa. Likewise, XynH2 was identified as xylanase XynF1 with a molecular weight of 35.402 kDa. Sequence analysis indicated that XynH1 belongs to glycosyl hydrolases family 10. The specific activity of XynH1 was measured at 476.9 U/mg. Optimal xylanase activity was observed at pH 6.0, and enzyme remained active within pH 4.0-10.0 and at a temperature below 70 °C. Mg(2+), Mn(2+), Ca(2+), and K(+) enhanced the XynH1 xylanase activity to 146, 122, 114, and 108%, respectively. XynH1 hydrolyzed Birchwood xylan and Larchwood xylan effectively. The K m and V max of XynH1 values determined were 1.16 mM and 336 μmol/min/mg with Birchwood xylan as the substrate. A. oryzae HML366 xylanase XynH1 showed superior heat and pH tolerance, therefore may have significant applications in paper and biofuel industries. These studies constitute the first investigation of the xylanase activities of the hypothetical protein XP_001826985.1 form A. oryzae.

  17. Improvement of Xylanase Production by Cochliobolus sativus in Submerged Culture

    Directory of Open Access Journals (Sweden)

    Yasser Bakri

    2008-01-01

    Full Text Available The xylanase production by a new Cochliobolus sativus Cs5 strain was improved under submerged fermentation. The xylanase was induced by xylan and repressed by glucose, sucrose, maltose, xylose, starch and cellulose. Highest enzyme production (98.25 IU/mL was recorded when wheat straw (4 % by mass per volume was used as a carbon source after 120 h of incubation. NaNO3 increased xylanase production 5.4-fold as compared to the control. Optimum initial pH was found to be 4.5 to 5. The C. sativus Cs5 strain grown under submerged culture in a simple medium proved to be a promising microorganism for xylanase production.

  18. Biocontrol Potential of Steinernema thermophilum and Its Symbiont Xenorhabdus indica Against Lepidopteran Pests: Virulence to Egg and Larval Stages.

    Science.gov (United States)

    Kalia, Vinay; Sharma, Garima; Shapiro-Ilan, David I; Ganguly, Sudershan

    2014-03-01

    Under laboratory conditions, the biocontrol potential of Steinernema thermophilum was tested against eggs and larval stages of two important lepidopteran insect pests, Helicoverpa armigera and Spodoptera litura (polyphagous pests), as well as Galleria mellonella (used as a model host). In terms of host susceptibility of lepidopteran larvae to S. thermophilum, based on the LC50 36 hr after treatment, G. mellonella (LC50 = 16.28 IJ/larva) was found to be more susceptible than S. litura (LC50 = 85 IJ/larva), whereas neither host was found to be significantly different from H. armigera (LC50 = 54.68 IJ/larva). In addition to virulence to the larval stages, ovicidal activity up to 84% was observed at 200 IJ/50 and 100 eggs of H. armigera and S. litura, respectively. To our knowledge this is the first report of entomopathogenic nematode pathogenicity to lepidopteran eggs. Production of infective juvenile (IJ) nematodes/insect larva was also measured and found to be positively correlated with rate of IJ for H. armigera (r = 0.990), S. litura (r = 0.892), as well as G. mellonella (r = 0.834). Both Phase I and Phase II of symbiotic bacteria Xenorhabdus indica were tested separately against neonates of H. armigera and S. litura by feeding assays and found to be virulent to the target pests; phase variation did not affect the level of virulence. Thus S. thermophilum as well as the nematode's symbiotic bacteria applied separately have the potential to be developed as biocontrol agents for key lepidopteran pests.

  19. A highly thermostable alkaline cellulase-free xylanase from thermoalkalophilic Bacillus sp. JB 99 suitable for paper and pulp industry: purification and characterization.

    Science.gov (United States)

    Shrinivas, Dengeti; Savitha, Gunashekaran; Raviranjan, Kumar; Naik, Gajanan Ramchandra

    2010-11-01

    A highly thermostable alkaline xylanase was purified to homogeneity from culture supernatant of Bacillus sp. JB 99 using DEAE-Sepharose and Sephadex G-100 gel filtration with 25.7-fold increase in activity and 43.5% recovery. The molecular weight of the purified xylanase was found to be 20 kDA by SDS-PAGE and zymogram analysis. The enzyme was optimally active at 70 °C, pH 8.0 and stable over pH range of 6.0-10.0.The relative activity at 9.0 and 10.0 were 90% and 85% of that of pH 8.0, respectively. The enzyme showed high thermal stability at 60 °C with 95% of its activity after 5 h. The K (m) and V (max) of enzyme for oat spelt xylan were 4.8 mg/ml and 218.6 µM min(-1) mg(-1), respectively. Analysis of N-terminal amino acid sequence revealed that the xylanase belongs to glycosyl hydrolase family 11 from thermoalkalophilic Bacillus sp. with basic pI. Substrate specificity showed a high activity on xylan-containing substrate and cellulase-free nature. The hydrolyzed product pattern of oat spelt xylan on thin-layer chromatography suggested xylanase as an endoxylanase. Due to these properties, xylanase from Bacillus sp. JB 99 was found to be highly compatible for paper and pulp industry.

  20. Trichoderma reesei xylanase 5 is defective in the reference strain QM6a but functional alleles are present in other wild-type strains.

    Science.gov (United States)

    Ramoni, Jonas; Marchetti-Deschmann, Martina; Seidl-Seiboth, Verena; Seiboth, Bernhard

    2017-05-01

    Trichoderma reesei is a paradigm for the regulation and industrial production of plant cell wall-degrading enzymes. Among these, five xylanases, including the glycoside hydrolase (GH) family 11 XYN1 and XYN2, the GH10 XYN3, and the GH30 XYN4 and XYN6, were described. By genome mining and transcriptome analysis, a further putative xylanase, encoded by xyn5, was identified. Analysis of xyn5 from the genome-sequenced reference strain T. reesei QM6a shows that it encodes a non-functional, truncated form of XYN5. However, non-truncated orthologues are present in other genome sequenced Trichoderma spp., and sequencing of xyn5 in other T. reesei wild-type isolates shows that they harbor a putative functional xyn5 allele. In silico analysis and 3D modeling revealed that the encoded XYN5 has significant structural similarities to xylanases of the GH11 family, including a GH-typical substrate binding groove and a carboxylate pair in the active site. The xyn5 of wild-type strain TUCIM1282 was recombinantly expressed in a T. reesei strain with a (hemi)cellulase-free background and the corresponding protein purified to apparent homogeneity. The pH and temperature optima and the kinetic parameters of the purified XYN5 were pH 4, 50 °C, and V max  = 2646 nkat/mg with a K m of 9.68 mg/ml. This functional xyn5 allele was used to replace the mutated version which led to an overall increase of the xylanolytic activity. These findings are of particular importance as GH11 xylanases are of high biotechnological relevance, and T. reesei is one of the main industrial producers of such lignocellulose-degrading enzymes.

  1. Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate

    Science.gov (United States)

    Beitel, Susan Michelz; Fortkamp, Diana; Terrasan, César Rafael Fanchini; de Almeida, Alex Fernando

    2013-01-01

    In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn2+ and the reducing agents β-mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg2+, Zn2+, and Cu2+ as well as the detergent SDS were strong inhibitors of the enzyme. The use of brewer's spent grain as substrate for xylanase production cannot only add value and decrease the amount of this waste but also reduce the xylanase production cost. PMID:23762855

  2. Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate

    Directory of Open Access Journals (Sweden)

    Adriana Knob

    2013-01-01

    Full Text Available In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn2+ and the reducing agents β-mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg2+, Zn2+, and Cu2+ as well as the detergent SDS were strong inhibitors of the enzyme. The use of brewer's spent grain as substrate for xylanase production cannot only add value and decrease the amount of this waste but also reduce the xylanase production cost.

  3. Fine-Scale Biogeographical Boundary Delineation and Sub-population Resolution in the Symbiodinium thermophilum Coral Symbiont Group From the Persian/Arabian Gulf and Gulf of Oman

    KAUST Repository

    Hume, Benjamin C. C.

    2018-04-24

    The adaptation of tropical coral communities to the world\\'s hottest sea, the Persian/Arabian Gulf (PAG), has recently been associated with ecological selection acting on a group of coral-associated algal symbionts, the Symbiodinium thermophilum group. Previous studies have shown that considerable genetic diversity exists within the group and that group members found within the PAG are significantly differentiated from those found externally, in the Gulf of Oman and wider waters. However, little is known about this genetic diversity. As an initial step towards understanding whether this diversity could represent niche adapted, selectable populations within the S. thermophilum group that may act as natural sources of stress tolerant associations to Indo-Pacific reefs, we investigate whether the diversity is structured between populations and where the location of the internal-external genetic partition lies. We use regions of the nuclear ribosomal DNA (ITS1-5.8S-ITS2) and chloroplastic psbA gene (non-coding region) from >100 S. thermophilum group-harbouring Porites spp. (P. lobata, P. lutea, and P. harrisoni) sampled across steep temperature and salinity gradients to conduct analyses of variance and create maximum parsimony networks to assess genetic structure and (dis)similarity within and between populations of S. thermophilum found within the PAG and externally in the Gulf of Oman. Our analyses resolve a sharp genetic boundary between Symbiodinium populations in the western Strait of Hormuz and identify significant genetic structure between populations with as little as 20 km between them demonstrating that differentiation between populations is likely due to factors other than limited connectivity. Further, we hypothesize that genotypes identified outside of the PAG in the Gulf of Oman existing in near-oceanic salinities, yet thermally challenging waters, putatively represent candidates for stress-tolerant symbionts that could act as natural seed populations of

  4. Biotechnology of microbial xylanases: enzymology, molecular biology, and application.

    Science.gov (United States)

    Subramaniyan, S; Prema, P

    2002-01-01

    Xylanases are hydrolases depolymerizing the plant cell wall component xylan, the second most abundant polysaccharide. The molecular structure and hydrolytic pattern of xylanases have been reported extensively and the mechanism of hydrolysis has also been proposed. There are several models for the gene regulation of which this article could add to the wealth of knowledge. Future work on the application of these enzymes in the paper and pulp, food industry, in environmental science, that is, bio-fueling, effluent treatment, and agro-waste treatment, etc. require a complete understanding of the functional and genetic significance of the xylanases. However, the thrust area has been identified as the paper and pulp industry. The major problem in the field of paper bleaching is the removal of lignin and its derivatives, which are linked to cellulose and xylan. Xylanases are more suitable in the paper and pulp industry than lignin-degrading systems.

  5. Production of xylanases by an Aspergillus niger strain in wastes grain

    Directory of Open Access Journals (Sweden)

    Simone Cristine Izidoro

    2014-08-01

    Full Text Available Many fungi are used in order to extract products from their metabolism through bioprocesses capable of minimizing adverse effects caused by agro-industrial wastes in the environment. The aim of this study was to evaluate the xylanase production by an Aspergillus niger strain, using agro-industrial wastes as substrate. Brewer’s spent grain was the best inducer of xylanase activity. Higher levels of xylanase were obtained when the fungus was grown in liquid Vogel medium, pH 5.0, at 30ºC, during 5 days. The temperature for optimum activity was 50ºC and optimum pH 5.0. The enzyme was stable at 50ºC, with a half-life of 240 min. High pH stability was verified from pH 4.5 to 7.0. These characteristics exhibited by A. niger xylanase turn this enzyme attractive for some industrial applications, such as in feed and food industries. Additionally, the use of brewer’s spent grain, an abundantly available and low-cost residue, as substrate for xylanase production can not only add value and decrease the amount of this waste, but also reduce xylanase production cost.

  6. Xylanases of marine fungi of potential use for biobleaching of paper pulp

    Digital Repository Service at National Institute of Oceanography (India)

    Raghukumar, C.; Muraleedharan, U.; Gaud, V.R.; Mishra, R.

    isolates obtained from marine habitat showed alkaline xylanase activity. The crude enzyme from NIOCC isolate # 3 (Aspergillus niger) with high xylanase activity, cellulase-free and unique properties containing 580 U L-1 of xylanase, could bring about...

  7. Developing a xylanase XYNZG from Plectosphaerella cucumerina for baking by heterologously expressed in Kluyveromyces lactis.

    Science.gov (United States)

    Zhan, Fei Xiang; Wang, Qin Hong; Jiang, Si Jing; Zhou, Yu Ling; Zhang, Gui Min; Ma, Yan He

    2014-12-16

    Xylanase can replace chemical additives to improve the volume and sensory properties of bread in the baking. Suitable baking xylanase with improved yield will promote the application of xylanase in baking industry. The xylanase XYNZG from the Plectosphaerella cucumerina has been previously characterized by heterologous expression in Pichia pastoris. However, P. pastoris is not a suitable host for xylanase to be used in the baking process since P. pastoris does not have GRAS (Generally Regarded As Safe) status and requires large methanol supplement during the fermentation in most conditions, which is not allowed to be used in the food industry. Kluyveromyces lactis, as another yeast expression host, has a GRAS status, which has been successfully used in food and feed applications. No previous work has been reported concerning the heterologous expression of xylanase gene xynZG in K. lactis with an aim for application in baking. The xylanase gene xynZG from the P. cucumerina was heterologously expressed in K. lactis. The recombinant protein XYNZG in K. lactis presented an approximately 19 kDa band on SDS-PAGE and zymograms analysis. Transformant with the highest halo on the plate containing the RBB-xylan (Remazol Brilliant Blue-xylan) was selected for the flask fermentation in different media. The results indicated that the highest activity of 115 U/ml at 72 h was obtained with the YLPU medium. The mass spectrometry analysis suggested that the hydrolytic products of xylan by XYNZG were mainly xylobiose and xylotriose. The results of baking trials indicated that the addition of XYNZG could reduce the kneading time of dough, increase the volume of bread, improve the texture, and have more positive effects on the sensory properties of bread. Xylanase XYNZG is successfully expressed in K. lactis, which exhibits the highest activity among the published reports of the xylanase expression in K. lactis. The recombinant XYNZG can be used to improve the volume and sensory

  8. Thermo-mechanical and micro-structural properties of xylanase containing whole wheat bread

    Directory of Open Access Journals (Sweden)

    G. Ghoshal

    2016-12-01

    Full Text Available Xylanase is a hemicellulase that can hydrolyses the complex polysaccharides. Hemicelluloses are main components of cell walls of cereal grains. Moreover, hemicelluloses are considered as potential sources of mono- and oligosaccharides. In this study, influence of xylanase on the physicochemical properties and sensory qualities of the whole wheat bread during storage was investigated. Studies of whole wheat bread on microstructure, texture, thermotics, Scanning Electron Microscopic (SEM, X-Ray Diffraction (XRD were conducted at ambient temperature of 25 and 4 °C respectively. During storage at different temperatures, bread containing xylanase exhibited less firmness but larger volume with whiter crumb color and longer shelf life as compared to control bread. Results of firmness, enthalpy, Fourier Transformation Infra Red (FTIR and X-Ray Diffraction (XRD studies suggested a lower staling rate of bread containing xylanase as compared to control one. Bread containing xylanase showed a smoother surface and more uniform pore size than the control. Significant differences in microstructure of control and bread containing xylanase were observed which might be attributed due to the change in water starch gluten interaction. These differences were also found to be interrelated to the textural properties of bread. Better sensory features were achieved in bread containing xylanase.

  9. A novel halotolerant xylanase from marine isolate Bacillus subtilis cho40: gene cloning and sequencing

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Verma, P.; Deobagkar, D.

    A novel halotolerant xylanase from marine bacterium Bacillus subtilis cho40 isolated from Chorao island of Mandovi estuary Goa, India has been reported. Extracellular xylanase was produced by using agricultural residue such as wheat bran as carbon...

  10. The expression of a xylanase targeted to ER-protein bodies provides a simple strategy to produce active insoluble enzyme polymers in tobacco plants.

    Directory of Open Access Journals (Sweden)

    Immaculada Llop-Tous

    Full Text Available BACKGROUND: Xylanases deserve particular attention due to their potential application in the feed, pulp bleaching and paper industries. We have developed here an efficient system for the production of an active xylanase in tobacco plants fused to a proline-rich domain (Zera of the maize storage protein γ-zein. Zera is a self-assembling domain able to form protein aggregates in vivo packed in newly formed endoplasmic reticulum-derived organelles known as protein bodies (PBs. METHODOLOGY/PRINCIPAL FINDINGS: Tobacco leaves were transiently transformed with a binary vector containing the Zera-xylanase coding region, which was optimized for plant expression, under the control of the 35S CaMV promoter. The fusion protein was efficiently expressed and stored in dense PBs, resulting in yields of up to 9% of total protein. Zera-xylanase was post-translationally modified with high-mannose-type glycans. Xylanase fused to Zera was biologically active not only when solubilized from PBs but also in its insoluble form. The resistance of insoluble Zera-xylanase to trypsin digestion demonstrated that the correct folding of xylanase in PBs was not impaired by Zera oligomerization. The activity of insoluble Zera-xylanase was enhanced when substrate accessibility was facilitated by physical treatments such as ultrasound. Moreover, we found that the thermostability of the enzyme was improved when Zera was fused to the C-terminus of xylanase. CONCLUSION/SIGNIFICANCE: In the present work we have successfully produced an active insoluble aggregate of xylanase fused to Zera in plants. Zera-xylanase chimeric protein accumulates within ER-derived protein bodies as active aggregates that can easily be recovered by a simple density-based downstream process. The production of insoluble active Zera-xylanase protein in tobacco outlines the potential of Zera as a fusion partner for producing enzymes of biotechnological relevance. Zera-PBs could thus become efficient and low

  11. Molecular Cloning and Sequencing of AlkalophilicCellulosimicrobium cellulans CKMX1 Xylanase Gene Isolated from Mushroom Compost and Characterization of the Gene Product

    Directory of Open Access Journals (Sweden)

    Abhishek Walia

    2015-12-01

    Full Text Available ABSTRACT A xylanolytic bacterium was isolated from mushroom compost by using enrichment technique. Results from the metabolic fingerprinting, whole-cell fatty acids methyl ester analysis and 16S rDNA sequencing suggested the bacterium to be Cellulosimicrobium cellulans CKMX1. Due to the xylanolytic activity of this bacterium, isolation and characterization of the xylanase gene were attempted. A distinct fragment of about 1671 bp was successfully amplified using PCR and cloned into Escherichia coli DH5α. A BLAST search confirmed that the DNA sequence from the amplified fragment was endo-1, 4-beta-xylanase, which was a member of glycoside hydrolase family 11. It showed 98% homology withCellulosimicrobium sp. xylanase gene (Accession no. FJ859907.1 reported from the gut of Eisenia fetida in Korea. In silicophysico-chemical characterization of amino acid sequence of xylanase showed an open reading frame encoding a 556 amino acid sequence with a molecular weight of 58 kDa and theoretical isolectric point (pI of 4.46 was computed using Expasy's ProtParam server. Secondary and homology based 3D structure of xylanase was analysed using SOPMA and Swiss-Prot software.

  12. Statistical optimization of activity and stability of β-xylanase ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-10-20

    Oct 20, 2008 ... A factorial design was performed to find the best conditions of pH and temperature for β-xylanase activity and to maintain its activity for prolonged periods of time of pure xylanase produced by newly isolated Thermomyces lanuginosus THKU-49. The central composite design (CCD) used for the analysis of ...

  13. Optimization of biosynthesis conditions and catalitic behavior evaluation of cellulase-free xylanase produced by a new Streptomyces sp. strain

    Directory of Open Access Journals (Sweden)

    GABRIELA BAHRIM

    2011-07-01

    Full Text Available Cellulase-free xylanase by Streptomyces sp.P12-137 was obtained bycultivation on the wheat bran as the sole carbon source. The effect of carbon and nitrogen sources and a ratio of them on the cellulase-free xylanase production was investigated. The new isolate Streptomyces sp. strain was able to grow in submerged system and to produce an increased level of xylanase. Wheat bran induced xylanase biosynthesis yield at a high level (9.27 UA/ml. For economical reasons cultivation was achieved on a cheap fermentative medium represented by agro-industrial wastes. The optima of the pH and temperature of the crude xylanase activity were 5.5 and 70°C,respectively.

  14. Xylanase XYN IV from Trichoderma reesei showing exo- and endo-xylanase activity

    Science.gov (United States)

    A novel xylanase from Trichoderma reesei Rut C30, named XYN IV, was purified from the cellulolytic system of the fungus. The enzyme was discovered on its ability to attack aldotetraohexenuronic acid (HexA-2Xyl-4Xyl-4Xyl, HexA3Xyl3), releasing the reducing-end xylose residue. XYN IV exhibited catalyt...

  15. Characterization of Two Endo-β-1, 4-Xylanases from Myceliophthora thermophila and Their Saccharification Efficiencies, Synergistic with Commercial Cellulase

    Directory of Open Access Journals (Sweden)

    Abdul Basit

    2018-02-01

    Full Text Available The xylanases with high specific activity and resistance to harsh conditions are of high practical value for biomass utilization. In the present study, two new GH11 xylanase genes, MYCTH_56237 and MYCTH_49824, have been cloned from thermophilic fungus Myceliophthora thermophila and expressed in Pichia pastoris. The specific activities of purified xylanases reach approximately 1,533.7 and 1,412.5 U/mg, respectively. Based on multiple template-based homology modeling, the structures of their catalytic domains are predicted. Enzyme activity was more effective in 7.5 L fermentor, yielding 2,010.4 and 2,004.2 U/mL, respectively. Both enzymes exhibit optimal activity at 60°C with pH of 6.0 and 7.0, respectively. Their activities are not affected by EDTA and an array of metal ions. The kinetic constants have been determined for MYCTH_56237 (Km = 8.80 mg/mL, Vmax = 2,380 U/mg and MYCTH_49824 (Km = 5.67 mg/mL, Vmax = 1,750 U/mg. More importantly, both xylanases significantly cooperate with the commercial cellulase Celluclast 1.5 L in terms of the saccharification efficiency. All these biochemical properties of the xylanases offer practical potential for future applications.

  16. Effect of corn cobs concentration on xylanase biosynthesis by ...

    African Journals Online (AJOL)

    Corn cobs, an indigenous carbon source, were tested as substrate by Aspergillus niger for optimum synthesis of xylanase using the submerged fermentation technique. The trials for xylanase production were conducted at three concentration levels (2.5, 3.0 and 3.5%) of corn cobs, four different fermentation temperatures ...

  17. Production of cellulase-free xylanase by Aspergillus flavus: Effect of ...

    African Journals Online (AJOL)

    Nelciele

    Aspergillus flavus produced high levels of xylanase on agricultural residues with ... addition of 5% glycerol, mannitol or xylitol protected the xylanase from thermal inactivation at 50°C. The .... most often included in nutrient media for microbial.

  18. Synergistic effect of cellulase and xylanase during hydrolysis of natural lignocellulosic substrates.

    Science.gov (United States)

    Song, Hui-Ting; Gao, Yuan; Yang, Yi-Min; Xiao, Wen-Jing; Liu, Shi-Hui; Xia, Wu-Cheng; Liu, Zi-Lu; Yi, Li; Jiang, Zheng-Bing

    2016-11-01

    Synergistic combination of cellulase and xylanase has been performed on pre-treated substrates in many previous studies, while few on natural substrates. In this study, three unpretreated lignocellulosic substrates were studied, including corncob, corn stover, and rice straw. The results indicated that when the mixed cellulase and xylanase were applied, reducing sugar concentrations were calculated as 19.53, 15.56, and 17.35mg/ml, respectively, based on the 3,5 dinitrosalicylic acid (DNS) method. Compared to the treatment with only cellulose, the hydrolysis yields caused by mixed cellulase and xylanase were improved by 133%, 164%, and 545%, respectively. In addition, the conversion yield of corncob, corn stover, and rice straw by cellulase-xylanase co-treatment reached 43.9%, 48.5%, and 40.2%, respectively, based on HPLC analysis, which confirmed the synergistic effect of cellulase-xylanase that was much higher than either of the single enzyme treatment. The substrate morphology was also evaluated to explore the synergistic mechanism of cellulase-xylanase. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Solid-state Fermentation of Xylanase from Penicillium canescens 10-10c in a Multi-layer-packed Bed Reactor

    Science.gov (United States)

    Assamoi, Antoine A.; Destain, Jacqueline; Delvigne, Frank; Lognay, Georges; Thonart, Philippe

    Xylanase is produced by Penicillium canescens 10-10c from soya oil cake in static conditions using solid-state fermentation. The impact of several parameters such as the nature and the size of inoculum, bed-loading, and aeration is evaluated during the fermentation process. Mycelial inoculum gives more production than conidial inoculum. Increasing the quantity of inoculum enhances slightly xylanase production. Forced aeration induces more sporulation of strain and reduces xylanase production. However, forced moistened air improves the production compared to production obtained with forced dry air. In addition, increasing bed-loading reduces the specific xylanase production likely due to the incapacity of the Penicillium strain to grow deeply in the fermented soya oil cake mass. Thus, the best cultivation conditions involve mycelial inoculum form, a bed loading of 1-cm height and passive aeration. The maximum xylanase activity is obtained after 7 days of fermentation and attains 10,200 U/g of soya oil cake. These levels are higher than those presented in the literature and, therefore, show all the potentialities of this stock and this technique for the production of xylanase.

  20. Fine-Scale Biogeographical Boundary Delineation and Sub-population Resolution in the Symbiodinium thermophilum Coral Symbiont Group From the Persian/Arabian Gulf and Gulf of Oman

    KAUST Repository

    Hume, Benjamin C. C.; D'Angelo, Cecilia; Burt, John A.; Wiedenmann, Jö rg

    2018-01-01

    The adaptation of tropical coral communities to the world's hottest sea, the Persian/Arabian Gulf (PAG), has recently been associated with ecological selection acting on a group of coral-associated algal symbionts, the Symbiodinium thermophilum

  1. Xylanase-Aided Chlorine Dioxide Bleaching of Bagasse Pulp to Reduce AOX Formation

    Directory of Open Access Journals (Sweden)

    Yi Dai

    2016-02-01

    Full Text Available Xylanase pretreatment was used to improve the chlorine dioxide bleaching of bagasse pulp. The pulp was pretreated with xylanase, which was followed by a chlorine dioxide bleaching stage. The HexA content of the pulp and the AOX content of the bleaching effluent were measured using UV-Vis and GC-MS methods, respectively. The results showed that a good correlation occurred between HexA and kappa number. HexA content of the pulp decreased significantly after the xylanase pretreatment. The AOX content of the bleaching effluent decreased as HexA was removed from the pulp. It was found that AOX could be reduced by up to 29.8%, comparing XD0 with a D0 stage. Fourier transform infrared spectroscopy (FTIR was employed to determine the breakage of chemical bonds in the pulp. It revealed that some lignin and hemicellulose were removed after xylanase treatment. The GC-MS results showed that some toxic chloride such as 2,4,6-trichlorophenol could be completely removed after xylanase pretreatment.

  2. Development of a bifunctional xylanase-cellulase chimera with enhanced activity on rice and barley straws using a modular xylanase and an endoglucanase procured from camel rumen metagenome.

    Science.gov (United States)

    Khalili Ghadikolaei, Kamran; Akbari Noghabi, Kambiz; Shahbani Zahiri, Hossein

    2017-09-01

    The camel rumen metagenome is an untapped source of glycoside hydrolases. In this study, novel genes encoding for a modular xylanase (XylC) and a cellulase (CelC) were isolated from a camel rumen metagenome and expressed in Escherichia coli BL21 (DE3). XylC with xylanase (Xyn), CBM, and carbohydrate esterase (CE) domains was characterized as a β-1,4-endoxylanase with remarkable catalytic activity on oat-spelt xylan (K cat  = 2919 ± 57 s -1 ). The implication of XylC's modular structure in its high catalytic activity was analyzed by truncation and fusion construction with CelC. The resulting fusions including Cel-CBM, Cel-CBM-CE, and Xyn-CBM-Cel showed remarkable enhancement in CMCase activity with K cat values of 742 ± 12, 1289 ± 34.5, and 2799 ± 51 s -1 compared to CelC with a K cat of 422 ± 3.5 s -1 . It was also shown that the bifunctional Xyn-CBM-Cel with synergistic xylanase/cellulase activities was more efficient than XylC and CelC in hydrolysis of rice and barley straws.

  3. Production of high level of cellulase-free xylanase by the thermophilic fungus Thermomyces lanuginosus in laboratory and pilot scales using lignocellulosic materials

    Energy Technology Data Exchange (ETDEWEB)

    Gomes, J [Institute of Biotechnology, Technical Univ. of Graz (Austria); Purkarthofer, H [Institute of Biotechnology, Technical Univ. of Graz (Austria); Hayn, M [Institute of Biochemistry, Univ. of Graz (Austria); Kapplmueller, J [Voest-Alpine Industrieanlagen GmbH, Zellstofftechnik und Biomasseverwertung, Linz (Austria); Sinner, M [Voest-Alpine Industrieanlagen GmbH, Zellstofftechnik und Biomasseverwertung, Linz (Austria); Steiner, W [Institute of Biotechnology, Technical Univ. of Graz (Austria)

    1993-08-01

    Thermomyces lanuginosus, isolated from self-heated jute stacks in Bangladesh, was able to produce a very high level of cellulase-free xylanase in shake cultures using inexpensive lignocellulosic biomass. Of the nine lignocellulosic substrates tested, corn cobs were found to be the best inducer of xylanase activity. The laboratory results of xylanase production have been successfully scaled up to VABIO (Voest-Alpine Biomass Technology Center) scale using a 15-m[sup 3] fermentor for industrial production and application of xylanase. In addition, some properties of the enzyme in crude culture filtrate produced on corn cobs are presented. The enzyme exhibited very satisfactory storage stability at 4-30 C either as crude culture filtrate or as spray- or freeze-dried powder. The crude enzyme was active over a broad range of pH and had activity optima at pH 6.5 and 70-75 C. The enzyme was almost thermostable (91-92%) at pH 6.5 and 9.0 after 41 h preincubation at 55 C and lost only 20-33% activity after 188 h. In contrast, it was much less thermostable at pH 5.0 and 11.0 Xylanases produced on different lignocellulosic substrates exhibited differences in thermostability at 55 C and pH 6.5. (orig.)

  4. Enhancement of Cellulase and Xylanase Production Using pH-Shift and Dissolved Oxygen Control Strategy with Streptomyces griseorubens JSD-1.

    Science.gov (United States)

    Zhang, Dan; Luo, Yanqing; Chu, Shaohua; Zhi, Yuee; Wang, Bin; Zhou, Pei

    2016-01-01

    In this study, the production of cellulase and xylanase by Streptomyces griseorubens JSD-1 was improved by integrating the pH-shift and dissolved oxygen (DO)-constant control strategies. The pH-shift control strategy was carried out by analyzing the specific cell growth rate (μ) and specific enzyme formation rate (Q p) of S. griseorubens JSD-1. The pH was controlled at 8.0 during the first 48 h to maintain high cell growth, which then shifted to 7.5 after 48 h to improve the production of cellulase and xylanase. Using this method, the maximum activities of cellulase, xylanase, and filter paper enzyme (FPase) increased by 47.9, 29.5, and 113.6 %, respectively, compared to that obtained without pH control. On the basis of pH-shift control, the influence of DO concentrations on biomass and enzyme production was further investigated. The maximum production of cellulase, xylanase, and FPase reached 114.38 ± 0.96 U mL(-1), 330.57 ± 2.54 U mL(-1), and 40.11 ± 0.38 U mL(-1), which were about 1.6-fold, 0.6-fold, and 3.2-fold higher than that of neutral pH without DO control conditions. These results supplied a functional approach for improving cellulase and xylanase production.

  5. Agro-residues as Alternative for Xylanase Production by Filamentous Fungi

    Directory of Open Access Journals (Sweden)

    Adriana Knob

    2014-07-01

    Full Text Available Agro-industrial wastes are the most abundant renewable resource on earth and are available in large quantities. However, the disposal of these wastes presents an increasing environmental problem. Recently, there has been a great interest in the exploitation of these wastes as low-cost raw materials for the production of value-added compounds as microbial enzymes by submerged or solid-state fermentation systems. This review focuses on alternatives for xylanase production using agro-residues as substrates. In recent years, the interest in xylanase, which plays an important role in the breakdown of xylan, has markedly increased due to its wide variety of biotechnological applications. Among several agro-industrial residues that have been intensively investigated, many, such as wheat bran, wheat straw, and sugarcane bagasse, are suitable and result in high yields of xylanase, leading to low production costs. In addition, many relatively unexplored residues, such as oil palm wastes, sorghum straw, and coffee by-products, are some of the most promising substrates for xylanase production, requiring further assessment.

  6. Alkalistable endo-β-1,4-xylanase production from a newly isolated alkalitolerant Penicillium sp. SS1 using agro-residues.

    Science.gov (United States)

    Bajaj, Bijender Kumar; Sharma, Mukul; Sharma, Sunny

    2011-09-01

    Thermostable and alkalitolerant xylanases have got intense research focus due to their vast applications in various industries including pulp and paper, food, feed, textile, biofuel, etc. In the present investigation, a Penicillum sp. SS1 isolated from degrading woody material was found to produce moderately thermoactive and alkalistable endo-β-1,4-xylanase (xylanase). Maximum xylanase production was observed after fourth day of fermentation (43.84 IU/ml). The organism produced substantial quantities of xylanase using agricultural residues like wheat bran (20.6 IU/ml), rice bran (21.8 IU/ml) and sawdust (10.7 IU/ml) as carbon sources. The enzyme preparation was totally free of filter paper activity (FPase) and possessed negligible carboxymethyl cellulase (CMCase) activity; this could be an important feature of enzyme if the intended application of enzyme is in pulp and paper industries. Among nitrogen sources examined, yeast extract supported maximum xylanase production (45.74 IU/ml), and was followed by soybean meal (22.2 IU/ml) and ammonium sulphate (20 IU/ml). Maximum xylanase production was observed at initial medium pH 9 (25.6 IU/ml); however, at pH 8 and 10 also significantly high enzyme titre was observed (24 and 21.2 IU/ml, respectively). Thus, Penicillium sp. SS1 displayed capability of growing and producing xylanase at high alkaline pH (8-10). Maximum xylanase activity was reported at 50 °C, however, significantly high activity was observed at 60 °C (65.4%), however, at 70-80 °C activity was lost considerably. At 50-60 °C the enzyme retained very high activity up to 30-60 min (91-100%), however, prolonged incubation (90 min) caused considerable activity reduction (residual activity 63-68%).

  7. Alteration of white-rot basidiomycetes cellulase and xylanase activities in the submerged co-cultivation and optimization of enzyme production by Irpex lacteus and Schizophyllum commune.

    Science.gov (United States)

    Metreveli, Eka; Kachlishvili, Eva; Singer, Steven W; Elisashvili, Vladimir

    2017-10-01

    Mono and dual cultures of four white-rot basidiomycete species were evaluated for cellulase and xylanase activity under submerged fermentation conditions. Co-cultivation of Pycnoporus coccineus or Trametes hirsuta with Schizophyllum commune displayed antagonistic interactions resulting in the decrease of endoglucanase and total cellulase activities. In contrast, increases in cellulase and xylanase activity were revealed through the compatible interactions of Irpex lacteus with S. commune. Co-cultivation conditions were optimized for maximum enzyme production by I. lacteus and S. commune, the best producers of cellulase/xylanase and β-glucosidase, respectively. An optimized medium for the target enzyme production by the mixed culture was established in a laboratory fermenter yielding 7U/mL total cellulase, 142U/mL endoglucanase, 104U/mL xylanase, and 5.2U/mL β-glucosidase. The dual culture approach resulted in an enzymatic mixture with 11% improved lignocellulose saccharification potential compared to enzymes from a monoculture of I. lacteus. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Production of beta-xylanase and beta-xylosidase by the extremely halophilic archaeon Halorhabdus utahensis

    DEFF Research Database (Denmark)

    Wainø, M.; Ingvorsen, K.

    2003-01-01

    -xylosidase stabilities, approximately 55% and 83% of the initial beta-xylanase and beta-xylosidase activities, respectively, remained after 24 h incubation at 20% NaCl. The enzymes were also shown to be slightly thermophilic: P-xylanase activity exhibiting two optima at 55degrees and 70degreesC, while beta......The extremely halophilic archaeon, Halorhabdus utahensis, isolated from the Great Salt Lake, Utah, produced beta-xylanase and beta-xylosidase activities. Both enzymes were active over a broad NaCl range from near zero to 30% NaCl when tested with culture broth. A broad NaCl optimum was observed...... for beta-xylanase activity between 5% and 15% NaCl, while beta-xylosidase activity was highest at 5% NaCl. Almost half of the maximum activities remained at 27%-30% NaCl for both enzyme activities. When dialyzed culture supernatant and culture broth were employed for determination of beta-xylanase and beta...

  9. Induction and catabolite repression of cellulase and xylanase synthesis in the selected white-rot basidiomycetes

    Directory of Open Access Journals (Sweden)

    Aza Kobakhidze

    2016-09-01

    Full Text Available This paper reports regulation of endoglucanase (EC 3.2.1.4 and xylanase (EC 3.2.1.8 production in submerged cultivation of four white-rot basidiomycetes. Among carbon sources tested, the Avicel-based medium provided the highest levels of both hydrolases activities in all fungal cultures. However, the maximum endoglucanase and xylanase activities of the tested basidiomycetes varied from 3.9 U/ml and 7.4 U/ml in Fomes fomentarius to 34.2 U/ml and 29.5 U/ml in Pseudotrametes gibbosa, respectively (P. gibbosa specific cellulase and xylanase activities achieved 8.55 and 7.38 U/mg, respectively. Replacement of Avicel in the medium with carboxymethyl cellulose or xylan significantly lowered the enzyme yield of the tested fungi. Moreover, xylan did not ensure high xylanase activity of these fungi. Lignocellulosic substrates used as a carbon source provided poorer productivity (the specific CMCase activity was 1.12–3.62 U/mg and the specific xylanase activity was 1.95–3.32 U/mg. Expression of endoglucanase and xylanase synthesis in Panus lecometei and P. gibbosa was inducible; supplementation of the glycerol-containing medium with Avicel accompanied with a sharp increase of the fungal specific CMCase and xylanase activities from 0.02–0.04 U/mg to 1.30–8.55 U/mg. Supplementation of the Avicel-induced cultures with glucose or glycerol caused a catabolite repression of the cellulase and xylanase formation by P. gibbosa and P. lecometei. The enzyme synthesis resumed only after depletion of easily metabolizable carbon source, glucose or glycerol, from the medium. The data received suggest that in the tested fungi endoglucanase and xylanase synthesis is under control by a common regulatory mechanism.

  10. Production and properties of two types of xylanases from alkalophilic thermophilic Bacillus spp

    Energy Technology Data Exchange (ETDEWEB)

    Okazaki, W; Akahoshi, R; Akiba, T; Horikoshi, K

    1984-05-01

    Four strains (W1, W2, W3, and W4) of alkalophilic thermophilic bacteria which produced xylanase were isolated from soils. They were aerobic, spore-forming. Gram-positive, and rod-shaped bacteria and hence identified as the genus Bacillus. The optimal temperatures for growth of the four strains were between 45/sup 0/C and 50/sup 0/C and pH optima were between 9.0 and 10.0. No growth occurred below pH 7.0 or above 55/sup 0/C. The four strains produced xylanases in medium containing xylan or xylose under these conditions. The optimal pH and temperature for activities of the four xylanases ranged from 6.0 to 7.0 and from 65/sup 0/C to 70/sup 0/C, respectively. The four xylanases were stable in the wide pH range from 4.5 to 10.5 at 45/sup 0/C for 1 h. All xylanases split xylan to yield xylose and xylobiose.

  11. Isolation, Purification, and Characterization of Xylanase Produced by a New Species of Bacillus in Solid State Fermentation

    Directory of Open Access Journals (Sweden)

    Rajashri D. Kamble

    2012-01-01

    Full Text Available A thermoalkalophilic new species of Bacillus, similar to Bacillus arseniciselenatis DSM 15340, produced extracellular xylanase under solid state fermentation when wheat bran is used as carbon source. The extracellular xylanase was isolated by ammonium sulfate (80% precipitation and purified using ion exchange chromatography. The molecular weight of xylanase was ~29.8 ;kDa. The optimum temperature and pH for the enzyme activity were 50°C and pH 8.0. The enzyme was active on birchwood xylan and little active on p-nitrophenyl xylopyranoside but not on Avicel, CMC, cellobiose, and starch, showing its absolute substrate specificity. For birchwood xylan, the enzyme gave a Km 5.26 ;mg/mL and Vmax 277.7 ;μmol/min/mg, respectively. In addition, the xylanase was also capable of producing high-quality xylo-oligosaccharides, which indicated its application potential not only in pulp biobleaching processes but also in the nutraceutical industry.

  12. Production of xylanases and cellulases by aspergillus fumigatus ms16 using crude lignocellulosic substrates

    International Nuclear Information System (INIS)

    Naseeb, S.; Sohai, M.; Ahmad, A.; Khan, S.A.

    2015-01-01

    Xylanolytic and cellulolytic potential of a soil isolate, Aspergillus fumigatus (MS16) was studied by growing it on a variety of lignocellulosics, purified cellulose and xylan supplemented media. It was noted that carboxymethyl cellulose, salicin and xylan induce the -glucosidase and xylanase, respectively production of endoglucanase. The study revealed that Aspergillus fumigatus (MS16) co-secretes xylanase and cellulase in the presence of xylan; the ratio of the two enzymes was influenced by the initial pH of the medium. The maximum titers of xylanase and cellulase were noted at initial pH of 5.0. Relatively higher titers of both the enzymes were obtained when the fungus was cultivated at 35 degree C. Whereas, cellulase production was not detected when the fungus was cultivated at 40 degree C. The volumetric productivity (Qp) of xylanase was much higher than cellulases. The organism produced 2-3 folds higher titers of xylanase when grown on lignocellulosic materials in submerged cultivation than under solid-state cultivation, suggesting a different pattern of enzyme production in presence and in absence of free water. The partial characterization of enzymes showed that xylanase from this organism has -glucosidase. The higher melting temperature than endoglucanase and optimum temperature for activity was higher for xylanases than cellulases, whereas the optimum pH differed slightly i.e. in the range of 4.0-5.0. Enzyme preparation from this organism was loaded on some crude substrates and it showed that the enzyme preparation can be used to hydrolyze a variety of vegetable and agricultural waste materials. (author)

  13. Production of D-xylanases by thermophilic fungi using different methods of culture

    Energy Technology Data Exchange (ETDEWEB)

    Grajek, W

    1986-01-01

    Seven thermophilic strains of fungi were examined for their ability to produce D-xylanase in liquid and solid-state fermentations. It was confirmed that the best producers of xylanase, among microorganisms used, were H. lanuginosa and S. thermophile in liquid fermentation, and T. aurantiacus and H. lanuginosa in solid-state fermentations. The higher productivity of xylanase, namely 18,72 IU/ml, was obtained in liquid culture of H. lanuginosa. The pH and temperature optima of enzymes from liquid and solid-state cultures of fungi used were also presented.

  14. Characterization of the arabinoxylan-degrading machinery of the thermophilic bacterium Herbinix hemicellulosilytica-Six new xylanases, three arabinofuranosidases and one xylosidase.

    Science.gov (United States)

    Mechelke, M; Koeck, D E; Broeker, J; Roessler, B; Krabichler, F; Schwarz, W H; Zverlov, V V; Liebl, W

    2017-09-10

    Herbinix hemicellulosilytica is a newly isolated, gram-positive, anaerobic bacterium with extensive hemicellulose-degrading capabilities obtained from a thermophilic biogas reactor. In order to exploit its potential as a source for new industrial arabinoxylan-degrading enzymes, six new thermophilic xylanases, four from glycoside hydrolase family 10 (GH10) and two from GH11, three arabinofuranosidases (1x GH43, 2x GH51) and one β-xylosidase (GH43) were selected. The recombinantly produced enzymes were purified and characterized. All enzymes were active on different xylan-based polysaccharides and most of them showed temperature-vs-activity profiles with maxima around 55-65°C. HPAEC-PAD analysis of the hydrolysates of wheat arabinoxylan and of various purified xylooligosaccharides (XOS) and arabinoxylooligosaccharides (AXOS) was used to investigate their substrate and product specificities: among the GH10 xylanases, XynB showed a different product pattern when hydrolysing AXOS compared to XynA, XynC, and XynD. None of the GH11 xylanases was able to degrade any of the tested AXOS. All three arabinofuranosidases, ArfA, ArfB and ArfC, were classified as type AXH-m,d enzymes. None of the arabinofuranosidases was able to degrade the double-arabinosylated xylooligosaccharides XA 2+3 XX. β-Xylosidase XylA (GH43) was able to degrade unsubstituted XOS, but showed limited activity to degrade AXOS. Copyright © 2017. Published by Elsevier B.V.

  15. A xylanase gene directly cloned from the genomic DNA of alkaline wastewater sludge showing application potential in the paper industry.

    Science.gov (United States)

    Zhao, Yanyu; Luo, Huiying; Meng, Kun; Shi, Pengjun; Wang, Guozeng; Yang, Peilong; Yuan, Tiezheng; Yao, Bin

    2011-09-01

    A xylanase gene, aws-2x, was directly cloned from the genomic DNA of the alkaline wastewater sludge using degenerated PCR and modified TAIL-PCR. The deduced amino acid sequence of AWS-2x shared the highest identity (60%) with the xylanase from Chryseobacterium gleum belonging to the glycosyl hydrolase GH family 10. Recombinant AWS-2x was expressed in Escherichia coli BL21 (DE3) and purified to electrophoretic homogeneity. The enzyme showed maximal activity at pH 7.5 and 55 °C, maintained more than 50% of maximal activity when assayed at pH 9.0, and was stable over a wide pH range from 4.0 to 11.0. The specific activity of AWS-2x towards hardwood xylan (beechwood and birchwood xylan) was significantly higher than that to cereal xylan (oat spelt xylan and wheat arabinoxylan). These properties make AWS-2x a potential candidate for application in the pulp and paper industry.

  16. Production and characterization of thermostable xylanase from ...

    African Journals Online (AJOL)

    ajl2

    2013-02-20

    Feb 20, 2013 ... produced from Trichoderma (Huitron et al., 2008). Only a ... around their colonies against a red background, were selected and .... Resistance to antibiotic ..... Xylanases of marine fungi potential use for biobleaching of paper.

  17. Purification and Characterization of Haloalkaline, Organic Solvent Stable Xylanase from Newly Isolated Halophilic Bacterium-OKH

    Science.gov (United States)

    Sanghvi, Gaurav; Jivrajani, Mehul; Patel, Nirav; Jivrajani, Heta; Bhaskara, Govinal Badiger; Patel, Shivani

    2014-01-01

    A novel, alkali-tolerant halophilic bacterium-OKH with an ability to produce extracellular halophilic, alkali-tolerant, organic solvent stable, and moderately thermostable xylanase was isolated from salt salterns of Mithapur region, Gujarat, India. Identification of the bacterium was done based upon biochemical tests and 16S rRNA sequence. Maximum xylanase production was achieved at pH 9.0 and 37°C temperature in the medium containing 15% NaCl and 1% (w/v) corn cobs. Sugarcane bagasse and wheat straw also induce xylanase production when used as carbon source. The enzyme was active over a range of 0–25% sodium chloride examined in culture broth. The optimum xylanase activity was observed at 5% sodium chloride. Xylanase was purified with 25.81%-fold purification and 17.1% yield. Kinetic properties such as Km and Vmax were 4.2 mg/mL and 0.31 μmol/min/mL, respectively. The enzyme was stable at pH 6.0 and 50°C with 60% activity after 8 hours of incubation. Enzyme activity was enhanced by Ca2+, Mn2+, and Mg2+ but strongly inhibited by heavy metals such as Hg2+, Fe3+, Ni2+, and Zn2+. Xylanase was found to be stable in organic solvents like glutaraldehyde and isopropanol. The purified enzyme hydrolysed lignocellulosic substrates. Xylanase, purified from the halophilic bacterium-OKH, has potential biotechnological applications. PMID:27350996

  18. Benefits from additives and xylanase during enzymatic hydrolysis of bamboo shoot and mature bamboo.

    Science.gov (United States)

    Li, Kena; Wang, Xiao; Wang, Jingfeng; Zhang, Junhua

    2015-09-01

    Effects of additives (BSA, PEG 6000, and Tween 80) on enzymatic hydrolysis of bamboo shoot and mature bamboo fractions (bamboo green, bamboo timber, bamboo yellow, bamboo node, and bamboo branches) by cellulases and/or xylanase were evaluated. The addition of additives was comparable to the increase of cellulase loadings in the conversion of cellulose and xylan in bamboo fractions. Supplementation of xylanase (1 mg/g DM) with cellulases (10 FPU/g DM) in the hydrolysis of bamboo fractions was more efficient than addition of additives in the production of glucose and xylose. Moreover, addition of additives could further increase the glucose release from different bamboo fractions by cellulases and xylanase. Bamboo green exhibited the lowest hydrolyzability. Almost all of the polysaccharides in pretreated bamboo shoot fractions were hydrolyzed by cellulases with the addition of additives or xylanase. Additives and xylanase showed great potential for reducing cellulase requirement in the hydrolysis of bamboo. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Secretome analysis of the thermophilic xylanase hyper-producer Thermomyces lanuginosus SSBP cultivated on corn cobs.

    Science.gov (United States)

    Winger, A M; Heazlewood, J L; Chan, L J G; Petzold, C J; Permaul, K; Singh, S

    2014-11-01

    Thermomyces lanuginosus is a thermophilic fungus known for its ability to produce industrially important enzymes including large amounts of xylanase, the key enzyme in hemicellulose hydrolysis. The secretome of T. lanuginosus SSBP was profiled by shotgun proteomics to elucidate important enzymes involved in hemicellulose saccharification and to characterise the presence of other industrially interesting enzymes. This study reproducibly identified a total of 74 proteins in the supernatant following growth on corn cobs. An analysis of proteins revealed nine glycoside hydrolase (GH) enzymes including xylanase GH11, β-xylosidase GH43, β-glucosidase GH3, α-galactosidase GH36 and trehalose hydrolase GH65. Two commercially produced Thermomyces enzymes, lipase and amylase, were also identified. In addition, other industrially relevant enzymes not currently explored in Thermomyces were identified including glutaminase, fructose-bisphosphate aldolase and cyanate hydratase. Overall, these data provide insight into the novel ability of a cellulase-free fungus to utilise lignocellulosic material, ultimately producing a number of enzymes important to various industrial processes.

  20. Improvement of the catalytic efficiency of a hyperthermophilic xylanase from Bispora sp. MEY-1.

    Directory of Open Access Journals (Sweden)

    Xiaoyu Wang

    Full Text Available Extremophilic xylanases have attracted great scientific and industrial interest. In this study, a GH10 xylanase-encoding gene, Xyl10E, was cloned from Bispora sp. MEY-1 and expressed in Pichia pastoris GS115. Deduced Xyl10E shares the highest identities of 62% and 57% with characterized family GH10 xylanases from Talaromyces leycettanus and Penicillium canescens (structure 4F8X, respectively. Xyl10E was most active at 93 to 95°C and pH 4.0, retained more than 75% or 48% of the initial activity when heated at 80°C or 90°C for 30 min, respectively, and hardly lost activity at pH 1.0 to 7.0, but was completely inhibited by SDS. Two residues, A160 and A161, located on loop 4, were identified to play roles in catalysis. Mutants A160D/E demonstrated higher affinity to substrate with lower Km values, while mutants A161D/E mainly displayed elevated Vmax values. All of these mutants had significantly improved catalytic efficiency. According to the molecular dynamics simulation, the mutation of A160E was able to affect the important substrate binding site Y204 and then improve the substrate affinity, and the mutation of A161D was capable of forming a hydrogen bond with the substrate to promote the substrate binding or accelerate the product release. This study introduces a highly thermophilic fungal xylanase and reveals the importance of loop 4 for catalytic efficiency.

  1. Substituting Both the N-Terminal and "Cord" Regions of a Xylanase from Aspergillus oryzae to Improve Its Temperature Characteristics.

    Science.gov (United States)

    Li, Chuang; Li, Jianfang; Wang, Rui; Li, Xueqing; Li, Jinping; Deng, Chao; Wu, Minchen

    2018-02-06

    To improve the temperature characteristics of AoXyn11A, a mesophilic glycoside hydrolase family (GHF) 11 xylanase from Aspergillus oryzae CICC40186, its N-terminal and "cord" regions were selected to be substituted by means of the computer-aided analysis and calculation. In brief, one mutant, named ATX11A 41 , possessing the lowest root-mean-square deviation (RMSD) value was designed based on the molecular dynamics (MD) simulation by substituting the N-terminal 41 amino acids of AoXyn11A with the corresponding 42 ones of pXYL11, a thermophilic GHF11 xylanase from Thermobifida fusca. On the basis of the primary structure alignment of pXYL11 with ATX11A 41 (or AoXyn11A), another mutant, named ATX11A 41/cord , was designed by substituting the cord region ( 93 GTYNPGSGG 101 ) of ATX11A 41 with the corresponding one ( 93 GTYRPTG 99 ) of pXYL11. Both mutant-encoding genes, ATx11A 41 and ATx11A 41/cord , were constructed as designed theoretically by a megaprimer PCR technique and were expressed in Pichia pastoris GS115. The specific activities of recombinant (re) AoXyn11A, ATX11A 41 , and ATX11A 41/cord were 2916.7, 2667.6, and 2457.0 U/mg, respectively. The analysis of temperature characteristics displayed that the temperature optimum (T opt ) of reATX11A 41 or reATX11A 41/cord was 65 °C, which was 15 °C higher than that of reAoXyn11A. The thermal inactivation half-life (t 1/2 ) values of reATX11A 41 and reATX11A 41/cord at 60 °C were 55 and 83 min, respectively, whereas that of reAoXyn11A was only 18 min at 50 °C. The melting temperature (T m ) values of reAoXyn11A, reATX11A 41 , and reATX11A 41/cord were 54.2, 66.7, and 71.9 °C, respectively. In conclusion, the above findings indicated that the substitution of both the N-terminal and cord regions of a mesophilic AoXyn11A greatly contributed to its improved temperature characteristics.

  2. Reuse of wastewater from pulp industry for the optimization of fungal xylanase production

    Directory of Open Access Journals (Sweden)

    Geisiany Maria de Queiroz-Fernandes

    2017-05-01

    Full Text Available The production of enzymes using agro-industrial waste is a low cost alternative for the reuse of byproducts, with the subsequent impact decrease on the environment. Current analysis produced xylanase using fungus Aspergillus niger, with two types of wastewater generated during the pulp chemical bleaching phase as inducers. Xylanase was produced by submerged liquid fermentation and factorial design optimized parameters that influence production (concentration of wastewater and production period. Initial culture conditions (pH, temperature and agitation were optimized independently. Alkaline wastewater was more effective than acidic wastewater for the induction of xylanase in optimized conditions: 50% of culture medium, 7-day production, 30°C, pH 6.0 and agitation at 160 rpm. Despite different results, acidic and alkaline wastewaters induced xylanase production by A. niger when employed in concentrations lower than or equal to 50% of culture medium and in the most optimal conditions described above. Alkaline wastewater is highlighted as the most efficient for such production.

  3. Cellulase and Xylanase Production from Three Isolates of Indigenous Endophytic Fungi

    Science.gov (United States)

    Yopi; Tasia, W.; Melliawati, R.

    2017-12-01

    Cellulases and hemicellulases have good potential to be used in energy production, in pulp, paper, textile industries, as well as in animal feed industries. Moreover, its utilization in food industries also cannot be ignored, among others, cellulase and xylanase roles in bakery, wine, and fruit and vegetables juice production. One of the potential enzyme source is endophytic fungi. Object of this study is to explore the potency of endophytic fungi isolated from medicinal plants as source of cellulolytic and xylanolytic enzymes. HL.47F.216 is endophytic fungi isolated from traditional medicinal plants ironwood tree was determined as xylanase producer. HL.51F.235 from pin-flower tree is cellulase producer, while CBN.6F.29 which produces both xylanase and cellulase is originated from Madagascar periwinkle. HL.47F.216 showed 2.5 cm in clear zone diameter and its xylanase activity was 0.262 U/mL with optimum condition pH 7 at 50°C. HL.51F.235 showed 2.4 cm clear zone diameter and 0.239 U/mL of cellulase activity at pH 5 and 70°C. CBN.6F.29 showed 2.8 cm and 0.394 U/mL (pH 5, 40°C) for its cellulase activity, while 2.3 cm and 0.439 U/mL (pH 8, 70°C) for its xylanase activity. Xylanase from HL.47F.216 and CBN.6F.29 showed low molecular masses of 20 kDa and 37-50 kDa, respectively. Molecular masses for cellulases from HL.51F.235 and CBN.6F.29 were 25 and 50 kDa for HL.51F.235 and 100 kDa for CBN.6F.29. Based on macroscopic and microscopic identification, fungal isolate CBN.6F.29 is a member of Class Coelomycetes, while HL.47F.216 was Acremonium sp. and HL.51F.235 was Aspergillus nigri.

  4. Xylanase production from marine derived Trichoderma pleuroticola 08ÇK001 strain isolated from Mediterranean coastal sediments.

    Science.gov (United States)

    Korkmaz, Melih N; Ozdemir, Sennur C; Uzel, Ataç

    2017-10-01

    Xylanases constitutes one the most important enzymes with diverse applications in different industries such as bioethanol production, animal feedstock production, production of xylo-oligosaccharides, baking industry, paper and pulp industry, xylitol production, fruit juice, and beer finishing, degumming, and agriculture. Currently, industrial xylanases are mainly produced by Aspergillus and Trichoderma members. Since the marine environments are less studied compared to terrestrial environments and harbors great microbial diversity we aimed to investigate the xylanase production of 88 marine-derived filamentous fungal strains. These strains are semi-quantitatively screened for their extracellular xylanase production and Trichoderma pleuroticola 08ÇK001 xylanase activity was further characterized. Optimum pH and temperature was determined as 5.0 and 50 °C, respectively. The enzyme preparation retained 53% of its activity at pH 5.0 after 1 h and have found resistant against several ions and compounds such as K + , Ba 2+ , Na + , β-mercaptoethanol, Triton X-100 and toluene. This study demonstrates that marine-derived fungal strains are prolific sources for xylanase production and presents the first report about the production and characterization of xylanase from a marine derived T. pleuroticola strain. The characteristics of T. pleuroticola 08ÇK001 xylanase activity indicate possible employment in some industrial processes such as animal feed, juice and wine industries or paper pulping applications. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Effects of structure and xylanase treatment of brewers' spent grain on performance and nutrient availability in broiler chickens.

    Science.gov (United States)

    Denstadli, V; Westereng, B; Biniyam, H G; Ballance, S; Knutsen, S H; Svihus, B

    2010-06-01

    1. A factorial (2 x 3) feeding trial was set up to investigate the effects of coarse or finely ground brewers' spent grain (BSG) and xylanase treatment, either with no xylanase, top-dressed with xylanase or pre-treated with xylanase. 2. The experimental diets shared the same basal formulation and were fed to male broiler chickens (Ross 308) housed in individual cages from 12 to 29 d of age. 3. Xylanase pre-treatment reduced the dietary concentration of arabinoxylan by 15-30%. Pellet durability increased when BSG was ground. 4. Feed utilisation was significantly higher (6%) when the birds were given coarse BSG rather than ground BSG, whereas there was no significant effect of enzyme treatment. Apparent metabolisable energy was unaffected by the dietary treatments. 5. The overall starch digestibility was high (99%), with no dietary differences, whereas ileal protein digestibility was low (57%). Xylanase top-dressing tended to improve ileal protein digestibility but, in general, xylanase treatment had no major effect on overall performance in male broilers given diets with BSG.

  6. Enzymic hydrolysis of xylans. I. A high xylanase and beta-xylosidase producing strain of Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Conrad, D.

    1981-01-01

    Aspergillus niger, strain 110.42 (CBS) was selected as a producer of high xylanolytic activities. The time course of xylanase and beta-xylosidase production as well as the effect of pH and temperature on the activity of these enzymes were studied. High-performance liquid chromatography analysis of the enzymic degradation of arabinoxylan showed a nearly complete conversion to pentose sugars. Aspects of using crude xylanase preparations for enzymic saccharification of xylans are discussed.

  7. The Use of Xylanases from Different Microbial Origin in Bread Baking and Their Effects on Bread Qualities

    Science.gov (United States)

    Al-Widyan, Omar; Khataibeh, Moayad H.; Abu-Alruz, Khaled

    Effects of xylanases on bread quality were examined. Enzymes used were endo-xylanase (EC 3.2.1.8) from different sources of microorganisms. Baked loaves were assessed for Loaves volume, colour and staling rate. Xylanases produced from rumen microorganisms M6 had clearly positive effects on loaf volume of bread as well as anti-firming potential. M3 (produced from Trichoderma longibrachiatum) improved crumb softness. The use of xylanase for breadmaking lowered firmness of bread crumb effectively compared with control loaf. It can be summarized that xylanases had significant positive effects on bread characteristics. In particular, they had advantage in retarding the staling rate of bread. It is recommended that the optimum dosage of enzymes, method of application in industrial scale especially with xylanase should be studied further in order to gain the great advantages of enzyme addition in breadmaking.

  8. Xylanases, nucleic acids encoding them and methods for making and using them

    Energy Technology Data Exchange (ETDEWEB)

    Gray, Kevin A.; Dirmeier, Richard

    2018-02-27

    The invention relates to enzymes having xylanase, mannanase and/or glucanase activity, e.g., catalyzing hydrolysis of internal .beta.-1,4-xylosidic linkages or endo-.beta.-1,4-glucanase linkages; and/or degrading a linear polysaccharide beta-1,4-xylan into xylose. Thus, the invention provides methods and processes for breaking down hemicellulose, which is a major component of the cell wall of plants, including methods and processes for hydrolyzing hemicelluloses in any plant or wood or wood product, wood waste, paper pulp, paper product or paper waste or byproduct. In addition, methods of designing new xylanases, mannanases and/or glucanases and methods of use thereof are also provided. The xylanases, mannanases and/or glucanases have increased activity and stability at increased pH and temperature.

  9. Xylanases, nucleic acids encoding them and methods for making and using them

    Science.gov (United States)

    Gray, Kevin A; Dirmeier, Reinhard

    2013-07-16

    The invention relates to enzymes having xylanase, mannanase and/or glucanase activity, e.g., catalyzing hydrolysis of internal .beta.-1,4-xylosidic linkages or endo-.beta.-1,4-glucanase linkages; and/or degrading a linear polysaccharide beta-1,4-xylan into xylose. Thus, the invention provides methods and processes for breaking down hemicellulose, which is a major component of the cell wall of plants, including methods and processes for hydrolyzing hemicelluloses in any plant or wood or wood product, wood waste, paper pulp, paper product or paper waste or byproduct. In addition, methods of designing new xylanases, mannanases and/or glucanases and methods of use thereof are also provided. The xylanases, mannanases and/or glucanases have increased activity and stability at increased pH and temperature.

  10. EFFECT OF XYLANASE ADDED TO A RYE-BASED DIET ON NUTRIENT UTILIZATION IN PIGS

    Directory of Open Access Journals (Sweden)

    Jaroslav Heger

    2012-02-01

    Full Text Available The effect of enzyme xylanase derived from Trichoderma longibrachiatum supplemented to a rye-based diet on apparent ileal digestibility of amino acids and non-starch polysaccharides constituting sugars was studied. Enzymes supplementation at 200 mg.kg−1 increased (P˂0.05 the digestibility of total amino acids from 67.1 to 70.8. When the dietary concentration of enzyme increased from 0 to 100 mg.kg-1, the ileal digestibility of the NSP constituents gradually increased as well. No further increase was observed with the supplementation level of 200 mg.kg-1. The improvement in the digestibility of arabinose and xylose (685%, P˂0.05 was much higher in comparison with remaining sugars (110%, P˂0.05. The apparent ileal digestibility of galactose was positively influenced by xylanase but it remained negative in all dietary treatments, presumably due to the high concentration of galactose in endogenous secretions. It is concluded that xylanase effectively degrades non-starch polysaccharides in upper digestive tract and marginally improves amino acid availability in young pigs.

  11. Xylanase production by a thermo-tolerant Bacillus species under solid-state and submerged fermentation

    Directory of Open Access Journals (Sweden)

    Uma Gupta

    2009-12-01

    Full Text Available Effects of xylose on xylanase production by a thermophilic Bacillus sp showed diverse patterns on corn cob (CC and wheat bran (WB as sole carbon sources in solid- state fermentation (SSF and submerged fermentation (SmF. Supplementation of these media with either mineral salt solution (MSS or yeast extract peptone (YEP also exerted variable effects. While under SSF, xylose stimulated xylanase synthesis by 44.01%, on wheat bran supplemented with MSS, it decreased the enzyme activity by 12.89% with YEP supplementation. In SmF, however the enzyme synthesis was stimulated by xylose on supplementation with both MSS and YEP by 41.38% and 27.47%, respectively. On corn cob under SSF, xylose repression was significant both with MSS (26.92% and YEP (23.90% supplementation. Repression by xylose also took place on corn cob and YEP (19.69% under SmF, while significant stimulation (28.55% was observed by MSS supplementation. The possible role of media composition and fermentation conditions in the regulation of xylanase synthesis by xylose is discussed.

  12. Phylogenetic diversity and environment-specific distributions of glycosyl hydrolase family 10 xylanases in geographically distant soils.

    Directory of Open Access Journals (Sweden)

    Guozeng Wang

    Full Text Available BACKGROUND: Xylan is one of the most abundant biopolymers on Earth. Its degradation is mediated primarily by microbial xylanase in nature. To explore the diversity and distribution patterns of xylanase genes in soils, samples of five soil types with different physicochemical characters were analyzed. METHODOLOGY/PRINCIPAL FINDINGS: Partial xylanase genes of glycoside hydrolase (GH family 10 were recovered following direct DNA extraction from soil, PCR amplification and cloning. Combined with our previous study, a total of 1084 gene fragments were obtained, representing 366 OTUs. More than half of the OTUs were novel (identities of <65% with known xylanases and had no close relatives based on phylogenetic analyses. Xylanase genes from all the soil environments were mainly distributed in Bacteroidetes, Proteobacteria, Acidobacteria, Firmicutes, Actinobacteria, Dictyoglomi and some fungi. Although identical sequences were found in several sites, habitat-specific patterns appeared to be important, and geochemical factors such as pH and oxygen content significantly influenced the compositions of xylan-degrading microbial communities. CONCLUSION/SIGNIFICANCE: These results provide insight into the GH 10 xylanases in various soil environments and reveal that xylan-degrading microbial communities are environment specific with diverse and abundant populations.

  13. Cloning and expression of an endo-1,4-β-xylanase from the coffee berry borer, Hypothenemus hampei

    Directory of Open Access Journals (Sweden)

    Padilla-Hurtado Beatriz

    2012-01-01

    Full Text Available Abstract Background The coffee berry borer, Hypothenemus hampei, reproduces and feeds exclusively on the mature endosperm of the coffee seed, which has a cell wall composed mainly of a heterogeneous mixture of hemicellulose polysaccharides, including arabinoxylans. Xylanases are digestive enzymes responsible for the degradation of xylan based polymers, hydrolyzing them into smaller molecules that are easier to assimilate by insects. We report the cloning, expression and enzymatic characterization of a xylanase gene that was identified in the digestive tract of the coffee berry borer. Methods The complete DNA sequence encoding a H. hampei xylanase (HhXyl was obtained using a genome walking technique in a cDNA library derived from the borer digestive tract. The XIP-I gene was amplified from wheat (Triticum aestivum variety Soisson. A Pichia pastoris expression system was used to express the recombinant form of these enzymes. The xylanase activity and XIP-I inhibitory activity was quantified by the 3,5-dinitrosalicylic (DNS. The biological effects of XIP-I on borer individuals were evaluated by providing an artificial diet enriched with the recombinant XIP-I protein to the insects. Results The borer xylanase sequence contains a 951 bp open reading frame that is predicted to encode a 317-amino acid protein, with an estimated molecular weight of 34.92 kDa and a pI of 4.84. Bioinformatic analysis revealed that HhXyl exhibits high sequence homology with endo-β-D-xylanases of Streptomyces bingchenggensis from glycosyl hydrolase 10 (GH10. The recombinant xylanase showed maximal activity at pH 5.5 and 37°C. XIP-I expressed as a recombinant protein inhibited HhXyl activity in vitro and caused individual H. hampei mortality in bioassays when included as a supplement in artificial diets. Conclusion A xylanase from the digestive tract of the coffee berry borer was identified and functionally characterized. A xylanase inhibitor protein, XIP-I, from wheat was

  14. 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 ...

  15. Homologue expression of a fungal endo-1,4-β-D- xylanase using ...

    African Journals Online (AJOL)

    Jane

    2011-03-07

    Mar 7, 2011 ... Hemicellulose is the second source of renewable organic carbon on earth, with a high potential for the recovery of ... SSF, solid-state fermentation; XE, xylanase extract. .... Active fractions were pooled, concentrated and.

  16. Efficient utilization of xylanase and lipase producing thermophilic ...

    African Journals Online (AJOL)

    Efficient utilization of xylanase and lipase producing thermophilic marine actinomycetes ( Streptomyces albus and Streptomyces hygroscopicus ) in the production of ecofriendly alternative energy from waste.

  17. Statistical optimization of thermo-alkali stable xylanase production from Bacillus tequilensis strain ARMATI

    Directory of Open Access Journals (Sweden)

    Ameer Khusro

    2016-07-01

    Conclusions: The cellulase-free xylanase showed an alkali-tolerant and thermo-stable property with potentially applicable nature at industrial scale. This statistical approach established a major contribution in enzyme production from the isolate by optimizing independent factors and represents a first reference on the enhanced production of thermo-alkali stable cellulase-free xylanase from B. tequilensis.

  18. Production and characterization of xylanase from Bacillus thermoalkalophilus grown on agricultural wastes

    Energy Technology Data Exchange (ETDEWEB)

    Rajaram, S.; Varma, A. (Jawaharlal Nehru Univ., New Delhi (India). School of Life Sciences)

    1990-10-01

    Bacillus thermoalkalophilus isolated from termite-infested mound soils of the semi-arid zones of India had the ability to produce good amounts of xylanase(s) from cheap agricultural wastes. Of the two hemicellulosic substrates tested, bagasse was found to be the better inducer for xylanase production. Alkali treatment of bagasse and rice husk had varied effects on enzyme production. The enzyme preparation had activity optima at 60deg C and 70deg C and a half-life of 60 min at 65deg C. The enzyme was stable for 24 h over a pH range of 4.0-6.0, while maximum activity was observed at pH 6.0-7.0. Enzyme production and activity were inhibited by the end-product of xylan hydrolysis, xylose. (orig.).

  19. Characterization of a novel xylanase gene from rumen content of Hu sheep.

    Science.gov (United States)

    Wang, Qian; Luo, Yang; He, Bo; Jiang, Lin-Shu; Liu, Jian-Xin; Wang, Jia-Kun

    2015-12-01

    A novel xylanase gene, xyn-lxy, was cloned from a metagenomic fosmid library, which was previously constructed from the rumen contents of Hu sheep and was functionally characterized in Escherichia coli. The open reading frame was composed of 1923 bp and encoded for 640 amino acids, including a catalytic domain of glycosyl hydrolase family 10 and carbohydrate-binding module 9. The gene showed 97 % identity with uncultured bacterium Contig1552 but low similarity with xylanases from known cellulolytic-degrading microorganisms in the rumen. The recombinant XYN-LXY showed a specific activity of 664.7 U mg(-1). The optimal temperature and pH of the enzyme were 50 °C and 6.0, respectively. Specifically, XYN-LXY was exclusively activated by Mn(2+) among all of the cations and reducing agents tested in this study. An enzymatic hydrolysis assay revealed that XYN-LXY degraded birchwood xylan into xylooligosaccharide with a low degree of polymerization. After incubation for 4 h, the concentration of the dominant product, xylobiose, was 2.297 ± 0.175 mg ml(-1) (74.07 % of total product) followed by xylose with a concentration of 0.656 ± 0.010 mg ml(-1) (21.14 % of total product). The XYN-LXY exhibited deep degradation effects on the xylan substrate, which were rarely observed with endo-xylanase, making it a promising candidate for industrial application, especially in biofuel production.

  20. Xylanase supplementation on enzymatic saccharification of dilute acid pretreated poplars at different severities

    Science.gov (United States)

    Chao Zhang; Xinshu Zhuang; Zhao Jiang Wang; Fred Matt; Franz St. John; J.Y. Zhu

    2013-01-01

    Three pairs of solid substrates from dilute acid pretreatment of two poplar wood samples were enzymatically hydrolyzed by cellulase preparations supplemented with xylanase. Supplementation of xylanase improved cellulose saccharification perhaps due to improved cellulose accessibility by xylan hydrolysis. Total xylan removal directly affected enzymatic cellulose...

  1. Role in pathogenesis of two endo-beta-1,4-xylanase genes from the vascular wilt fungus Fusarium oxysporum.

    Science.gov (United States)

    Gómez-Gómez, E; Ruíz-Roldán, M C; Di Pietro, A; Roncero, M I G; Hera, C

    2002-04-01

    A gene, xyl4, whose predicted amino acid sequence shows significant homology with family 11 xylanases, was identified from the tomato vascular wilt fungus Fusarium oxysporum f. sp. lycopersici. Expression of xyl4 is induced on oat spelt xylan as the carbon source, subject to carbon catabolite repression and preferentially expressed at alkaline ambient pH. Transcript levels of xyl4 on an inducing carbon source are differentially regulated by the nature and concentration of the nitrogen source. As shown by RT-PCR, xyl4 is expressed by F. oxysporum during the entire cycle of infection on tomato plants. Targeted inactivation of xyl4 and of xyl3, a previously identified gene of F. oxysporum f. sp. lycopersici encoding a family 10 xylanase, had no detectable effect on virulence on tomato plants, demonstrating that both genes are not essential for pathogenicity.

  2. Two new xylanases with different substrate specificities from the human gut bacterium Bacteroides intestinalis DSM 17393.

    Science.gov (United States)

    Hong, Pei-Ying; Iakiviak, Michael; Dodd, Dylan; Zhang, Meiling; Mackie, Roderick I; Cann, Isaac

    2014-04-01

    Xylan is an abundant plant cell wall polysaccharide and is a dominant component of dietary fiber. Bacteria in the distal human gastrointestinal tract produce xylanase enzymes to initiate the degradation of this complex heteropolymer. These xylanases typically derive from glycoside hydrolase (GH) families 10 and 11; however, analysis of the genome sequence of the xylan-degrading human gut bacterium Bacteroides intestinalis DSM 17393 revealed the presence of two putative GH8 xylanases. In the current study, we demonstrate that the two genes encode enzymes that differ in activity. The xyn8A gene encodes an endoxylanase (Xyn8A), and rex8A encodes a reducing-end xylose-releasing exo-oligoxylanase (Rex8A). Xyn8A hydrolyzed both xylopentaose (X5) and xylohexaose (X6) to a mixture of xylobiose (X2) and xylotriose (X3), while Rex8A hydrolyzed X3 through X6 to a mixture of xylose (X1) and X2. Moreover, rex8A is located downstream of a GH3 gene (xyl3A) that was demonstrated to exhibit β-xylosidase activity and would be able to further hydrolyze X2 to X1. Mutational analyses of putative active site residues of both Xyn8A and Rex8A confirm their importance in catalysis by these enzymes. Recent genome sequences of gut bacteria reveal an increase in GH8 Rex enzymes, especially among the Bacteroidetes, indicating that these genes contribute to xylan utilization in the human gut.

  3. Two New Xylanases with Different Substrate Specificities from the Human Gut Bacterium Bacteroides intestinalis DSM 17393

    KAUST Repository

    Hong, Pei-Ying

    2014-01-24

    Xylan is an abundant plant cell wall polysaccharide and is a dominant component of dietary fiber. Bacteria in the distal human gastrointestinal tract produce xylanase enzymes to initiate the degradation of this complex heteropolymer. These xylanases typically derive from glycoside hydrolase (GH) families 10 and 11; however, analysis of the genome sequence of the xylan-degrading human gut bacterium Bacteroides intestinalis DSM 17393 revealed the presence of two putative GH8 xylanases. In the current study, we demonstrate that the two genes encode enzymes that differ in activity. The xyn8A gene encodes an endoxylanase (Xyn8A), and rex8A encodes a reducing-end xylose-releasing exo-oligoxylanase (Rex8A). Xyn8A hydrolyzed both xylopentaose (X5) and xylohexaose (X6) to a mixture of xylobiose (X2) and xylotriose (X3), while Rex8A hydrolyzed X3 through X6 to a mixture of xylose (X1) and X2. Moreover, rex8A is located downstream of a GH3 gene (xyl3A) that was demonstrated to exhibit β-xylosidase activity and would be able to further hydrolyze X2 to X1. Mutational analyses of putative active site residues of both Xyn8A and Rex8A confirm their importance in catalysis by these enzymes. Recent genome sequences of gut bacteria reveal an increase in GH8 Rex enzymes, especially among the Bacteroidetes, indicating that these genes contribute to xylan utilization in the human gut.

  4. Two New Xylanases with Different Substrate Specificities from the Human Gut Bacterium Bacteroides intestinalis DSM 17393

    KAUST Repository

    Hong, Pei-Ying; Iakiviak, M.; Dodd, D.; Zhang, M.; Mackie, R. I.; Cann, I.

    2014-01-01

    Xylan is an abundant plant cell wall polysaccharide and is a dominant component of dietary fiber. Bacteria in the distal human gastrointestinal tract produce xylanase enzymes to initiate the degradation of this complex heteropolymer. These xylanases typically derive from glycoside hydrolase (GH) families 10 and 11; however, analysis of the genome sequence of the xylan-degrading human gut bacterium Bacteroides intestinalis DSM 17393 revealed the presence of two putative GH8 xylanases. In the current study, we demonstrate that the two genes encode enzymes that differ in activity. The xyn8A gene encodes an endoxylanase (Xyn8A), and rex8A encodes a reducing-end xylose-releasing exo-oligoxylanase (Rex8A). Xyn8A hydrolyzed both xylopentaose (X5) and xylohexaose (X6) to a mixture of xylobiose (X2) and xylotriose (X3), while Rex8A hydrolyzed X3 through X6 to a mixture of xylose (X1) and X2. Moreover, rex8A is located downstream of a GH3 gene (xyl3A) that was demonstrated to exhibit β-xylosidase activity and would be able to further hydrolyze X2 to X1. Mutational analyses of putative active site residues of both Xyn8A and Rex8A confirm their importance in catalysis by these enzymes. Recent genome sequences of gut bacteria reveal an increase in GH8 Rex enzymes, especially among the Bacteroidetes, indicating that these genes contribute to xylan utilization in the human gut.

  5. Optimization of Xylanase production from Penicillium sp.WX-Z1 by a two-step statistical strategy: Plackett-Burman and Box-Behnken experimental design.

    Science.gov (United States)

    Cui, Fengjie; Zhao, Liming

    2012-01-01

    The objective of the study was to optimize the nutrition sources in a culture medium for the production of xylanase from Penicillium sp.WX-Z1 using Plackett-Burman design and Box-Behnken design. The Plackett-Burman multifactorial design was first employed to screen the important nutrient sources in the medium for xylanase production by Penicillium sp.WX-Z1 and subsequent use of the response surface methodology (RSM) was further optimized for xylanase production by Box-Behnken design. The important nutrient sources in the culture medium, identified by the initial screening method of Placket-Burman, were wheat bran, yeast extract, NaNO(3), MgSO(4), and CaCl(2). The optimal amounts (in g/L) for maximum production of xylanase were: wheat bran, 32.8; yeast extract, 1.02; NaNO(3), 12.71; MgSO(4), 0.96; and CaCl(2), 1.04. Using this statistical experimental design, the xylanase production under optimal condition reached 46.50 U/mL and an increase in xylanase activity of 1.34-fold was obtained compared with the original medium for fermentation carried out in a 30-L bioreactor.

  6. Thermoactive cellulase-free xylanase production from alkaliphilic ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-01-04

    Jan 4, 2010 ... The enzymatic hydrolysis of xylan, a major hemicellulose ... agar (glucose replaced by 1.0% w/v Birch-Wood Xylan (BW-X, ..... Thermal stability was determined by preincubating the xylanase at pH 9.0; at 60.0°C for 1 h.

  7. Heterologous Expression of Xylanase II from Aspergillus usamii in Pichia pastoris

    OpenAIRE

    Zhou, Chenyan; Wang, Yongtao; Wu, Minchen; Wang, Wu; Li, Dongfeng

    2009-01-01

    To efficiently produce xylanase for food processing industry, a gene encoding xylanase II (XynII) from Aspergillus usamii has been cloned into the vector pPIC9K and integrated into the genome of Pichia pastoris KM71 by electroporation. By means of minimal dextrose (MD) plates and PCR, the recombinant P. pastoris strains (His+Muts) have been obtained. Activity assay and SDS-PAGE demonstrate that XynII was extracellularly expressed in P. pastoris with the induction of methanol. In shake flask c...

  8. Catalytic performance of corn stover hydrolysis by a new isolate Penicillium sp. ECU0913 producing both cellulase and xylanase.

    Science.gov (United States)

    Shi, Qian-Qian; Sun, Jie; Yu, Hui-Lei; Li, Chun-Xiu; Bao, Jie; Xu, Jian-He

    2011-07-01

    A fungal strain, marked as ECU0913, producing high activities of both cellulase and xylanase was newly isolated from soil sample collected near decaying straw and identified as Penicillium sp. based on internal transcribed spacer sequence homology. The cultivation of this fungus produced both cellulase (2.40 FPU/ml) and xylanase (241 IU/ml) on a stepwisely optimized medium at 30 °C for 144 h. The cellulase and xylanase from Penicillium sp. ECU0913 was stable at an ambient temperature with half-lives of 28 and 12 days, respectively. Addition of 3 M sorbitol greatly improved the thermostability of the two enzymes, with half-lives increased by 2.3 and 188-folds, respectively. Catalytic performance of the Penicillium cellulase and xylanase was evaluated by the hydrolysis of corn stover pretreated by steam explosion. With an enzyme dosage of 50 FPU/g dry substrate, the conversions of cellulose and hemicellulose reached 77.2% and 47.5%, respectively, without adding any accessory enzyme.

  9. Xylanase, protease and superdosing phytase interactions in broiler performance, carcass yield and digesta transit time

    Directory of Open Access Journals (Sweden)

    Tiago T. dos Santos

    2017-06-01

    Full Text Available The interaction of xylanase, protease and superdosing (1,500 FTU/kg phytase in a 2 × 2 × 2 factorial arrangement was studied in broilers fed sorghum-based diets. A total of 2,800 one-day-old unsexed Ross 308 chicks were housed in 56 pens with 50 birds per pen, with or without inclusion of xylanase, protease and phytase, totaling 8 treatments and 7 replicates per treatment. Body weight (BW and feed intake (FI were measured at 21 and 42 days of age, and mortality corrected feed conversion ratio (FCR was calculated for each period and cumulatively. Tibia ash and carcass yield were determined in 2 birds per replicate at 21 and 42 days of age, respectively. Digesta transit time was determined at 21, 28, 35 and 42 days of age using 5 birds per replicate. Results showed that superdosing phytase increased BW and FI at 42 days of age (P < 0.05 and xylanase improved FCR (P < 0.05. Xylanase and phytase also positively influenced carcass yield and breast weight, respectively. Overall, inclusion of superdosing phytase increased transit time when included in a diet containing xylanase, and no change with protease inclusion. In conclusion, the beneficial effects of xylanase, protease and superdosing phytase in broiler performance were not additive. This limitation is likely not related to the lack of efficacy of any one of the individual enzymes but to a limitation of the bird to respond additively to successive additions of enzymes.

  10. Emerging role of N- and C-terminal interactions in stabilizing (β/α8 fold with special emphasis on Family 10 xylanases

    Directory of Open Access Journals (Sweden)

    Amit Bhardwaj

    2012-09-01

    Full Text Available Xylanases belong to an important class of industrial enzymes. Various xylanases have been purified and characterized from a plethora of organisms including bacteria, marine algae, plants, protozoans, insects, snails and crustaceans. Depending on the source, the enzymatic activity of xylanases varies considerably under various physico-chemical conditions such as temperature, pH, high salt and in the presence of proteases. Family 10 or glycosyl hydrolase 10 (GH10 xylanases are one of the well characterized and thoroughly studied classes of industrial enzymes. The TIM-barrel fold structure which is ubiquitous in nature is one of the characteristics of family 10 xylanases. Family 10 xylanases have been used as a “model system” due to their TIM-barrel fold to dissect and understand protein stability under various conditions. A better understanding of structure-stability-function relationships of family 10 xylanases allows one to apply these governing molecular rules to engineer other TIM-barrel fold proteins to improve their stability and retain function(s under adverse conditions. In this review, we discuss the implications of N-and C-terminal interactions, observed in family 10 xylanases on protein stability under extreme conditions. The role of metal binding and aromatic clusters in protein stability is also discussed. Studying and understanding family 10 xylanase structure and function, can contribute to our protein engineering knowledge.

  11. EMERGING ROLE OF N- AND C-TERMINAL INTERACTIONS IN STABILIZING (β;/α8 FOLD WITH SPECIAL EMPHASIS ON FAMILY 10 XYLANASES

    Directory of Open Access Journals (Sweden)

    Amit Bhardwaj

    2012-09-01

    Full Text Available Xylanases belong to an important class of industrial enzymes. Various xylanases have been purified and characterized from a plethora of organisms including bacteria, marine algae, plants, protozoans, insects, snails and crustaceans. Depending on the source, the enzymatic activity of xylanases varies considerably under various physico-chemical conditions such as temperature, pH, high salt and in the presence of proteases. Family 10 or glycosyl hydrolase 10 (GH10 xylanases are one of the well characterized and thoroughly studied classes of industrial enzymes. The TIM-barrel fold structure which is ubiquitous in nature is one of the characteristics of family 10 xylanases. Family 10 xylanases have been used as a “model system” due to their TIM-barrel fold to dissect and understand protein stability under various conditions. A better understanding of structure-stability-function relationships of family 10 xylanases allows one to apply these governing molecular rules to engineer other TIM-barrel fold proteins to improve their stability and retain function(s under adverse conditions. In this review, we discuss the implications of N-and C-terminal interactions, observed in family 10 xylanases on protein stability under extreme conditions. The role of metal binding and aromatic clusters in protein stability is also discussed. Studying and understanding family 10 xylanase structure and function, can contribute to our protein engineering knowledge.

  12. Sequential and simultaneous strategies for biorefining of wheat straw using room temperature ionic liquids, xylanases and cellulases.

    Science.gov (United States)

    Husson, Eric; Auxenfans, Thomas; Herbaut, Mickael; Baralle, Manon; Lambertyn, Virginie; Rakotoarivonina, Harivoni; Rémond, Caroline; Sarazin, Catherine

    2018-03-01

    Sequential and simultaneous strategies for fractioning wheat straw were developed in combining 1-ethyl-3-methyl imidazolium acetate [C2mim][OAc], endo-xylanases from Thermobacillus xylanilyticus and commercial cellulases. After [C2mim][OAc]-pretreatment, hydrolysis catalyzed by endo-xylanases of wheat straw led to efficient xylose production with very competitive yield (97.6 ± 1.3%). Subsequent enzymatic saccharification allowed achieving a total degradation of cellulosic fraction (>99%). These high performances revealed an interesting complementarity of [C2mim][OAc]- and xylanase-pretreatments for increasing enzymatic digestibility of cellulosic fraction in agreement with the structural and morphological changes of wheat straw induced by each of these pretreatment steps. In addition a higher tolerance of endo-xylanases from T. xylaniliticus to [C2mim][AcO] until 30% v/v than cellulases from T. reesei was observed. Based on this property, a simultaneous strategy combining [C2mim][OAc]- and endo-xylanases as pretreatment in a one-batch produced xylose with similar yield than those obtained by the sequential strategy. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Ecofriendly application of cellulase and xylanase producing marine ...

    African Journals Online (AJOL)

    windows

    2012-06-05

    Jun 5, 2012 ... producing marine Streptomyces clavuligerus as enhancer in ... pretreatment of cellulase, xylanase and the combination of enzymes. ... Energy from biomass holds a promising scope under ... investment, simplification of the fermentation media, ... biodegradation of lignocellulosic residues and enhanced ...

  14. Inexpensive, rapid procedure for bulk purification of cellulase-free. beta. -1,4-D-xylanase of high specific activity

    Energy Technology Data Exchange (ETDEWEB)

    Tan, L.U.L.; Yu, E.K.C.; Louis-Seize, G.W.; Saddler, J.N.

    1987-01-01

    A process has been developed for the bulk purification of cellulase-free ..beta..-14-D-xylanase from the fungus Tirchoderma harzianum E58. The process involved the primary step of ultrafiltering the culture filtrate via a 10,000-molecular-weight cut-off membrane to separate the cellulase (retentate) and xylanase (permeate) fractions. The cellulase component was concentrated by 40- to 60-fold, resulting in an enzyme complex that could effectively hydrolyze high concentrations of cellulose and xylan to glucose and xylose. The xylanase was concentrated and solvent exchanged by adsorption to a cationic exchanger, SP-ZetaPrep 250, followed by elution with a pH change in the buffer to give a purified and concentrated xylanase complex dissolved in a low-salt buffer. The resultant xylanase system was pure by the criteria of sodium dodecyl sulfate polyacrylamide electrophoresis, had a very high specific activity of 2400 IU/mg protein, was virtually free of filter paper activity, and had a ratio of contaminating filter paper activity of 2 x 10/sup -6/. Approximately 3.3 g protein, which contained in excess of 7 x 10/sup 6/ IU xylanase activity was obtained from 17 L original culture filtrate. The process scheme was designed to facilitate scale-up to an industrial level of production.

  15. Cellulase-poor xylanases produced by Trichoderma reesei RUT C-30 on hemicellulose substrates

    Energy Technology Data Exchange (ETDEWEB)

    Gamerith, G.; Groicher, R. (Lenzing AG (Austria). Dept. of Research and Development); Zeilinger, S.; Herzog, P.; Kubicek, C.P. (Technische Univ., Vienna (Austria). Abt. fuer Mikrobielle Biochemie)

    1992-12-01

    Hemicellulose components from industrial viscose fibre production are characterized by a lower cellulose content than commerical xylan and the pressence of a carboxylic acid fraction originating from the alkaline degradation of carbohydrates during the process. This substrate, after neutralization, can be used by Trichoderma reesei RUT C-30 for the production of cellulase-poor xylanases, useful for the pulp and paper industry. The yields of xylanase ranged up to almost 400 units/ml, with a ratio of carboxymethylcellulase/xylanase of less than 0.015. This crude xylanase enzyme mixture was shown to be superior to that obtained on beech-wood xylan when used for bleaching and, particularly, upgrading of hard-wood chemical pulp by selective removal of the xylan components. Biochemical studies indicate that the low cellulase production by T. reesei grown on these waste hemicelluloses is the result of a combination of at least three factors: (a) The comparatively low content of cellulose in these hemicellulosic wastes, (b) the inhibitory action of the carboxylic acid fraction present in the hemicellulosic wastes on growth and sporulation of T. reesei, and (c) the use of a mycelial inoculum that is unable to initiate the atack on the cellulose components within the carbon source. (orig.).

  16. Xylanase, CM-cellulase and avicelase production by the thermophilic fungus Sporotrichum thermophile

    Energy Technology Data Exchange (ETDEWEB)

    Margaritis, A; Merchant, R; Yaguchi, M

    1983-01-01

    When wheat straw was used as C source, S. thermophile produced large amounts of xylanase extracellularly in addition to CM-cellulase and Avicelase. These enzymes were isolated by alcohol precipitation, desalting, and column chromatography. The molecular weights were estimated to be 25,0065,000 and 84,000 for xylanase, CM-cellulase, and Avicelase, respectively. Serine and threonine were the most abundant amino acids and these enzymes are very acidic proteins.

  17. Dissecting electrostatic interactions in Bacillus circulans xylanase through NMR-monitored pH titrations

    Energy Technology Data Exchange (ETDEWEB)

    McIntosh, Lawrence P., E-mail: mcintosh@chem.ubc.ca; Naito, Daigo; Baturin, Simon J.; Okon, Mark; Joshi, Manish D. [University of British Columbia, Department of Biochemistry and Molecular Biology, Department of Chemistry, and Michael Smith Laboratories, Life Sciences Centre (Canada); Nielsen, Jens E. [University College Dublin, School of Biomolecular and Biomedical Science, Centre for Synthesis and Chemical Biology, UCD Conway Institute (Ireland)

    2011-09-15

    NMR-monitored pH titration curves of proteins provide a rich source of structural and electrostatic information. Although relatively straightforward to measure, interpreting pH-dependent chemical shift changes to obtain site-specific acid dissociation constants (pK{sub A} values) is challenging. In order to analyze the biphasic titrations exhibited by the side chain {sup 13}C{sup {gamma}} nuclei of the nucleophilic Glu78 and general acid/base Glu172 in Bacillus circulans xylanase, we have revisited the formalism for the ionization equilibria of two coupled acidic residues. In general, fitting NMR-monitored pH titration curves for such a system will only yield the two macroscopic pK{sub A} values that reflect the combined effects of both deprotonation reactions. However, through the use of mutations complemented with ionic strength-dependent measurements, we are able to extract the four microscopic pK{sub Ai} values governing the branched acid/base equilibria of Glu78 and Glu172 in BcX. These data, confirmed through theoretical calculations, help explain the pH-dependent mechanism of this model GH11 xylanase by demonstrating that the kinetically determined pK{sub A} values and hence catalytic roles of these two residues result from their electrostatic coupling.

  18. Dissecting electrostatic interactions in Bacillus circulans xylanase through NMR-monitored pH titrations

    International Nuclear Information System (INIS)

    McIntosh, Lawrence P.; Naito, Daigo; Baturin, Simon J.; Okon, Mark; Joshi, Manish D.; Nielsen, Jens E.

    2011-01-01

    NMR-monitored pH titration curves of proteins provide a rich source of structural and electrostatic information. Although relatively straightforward to measure, interpreting pH-dependent chemical shift changes to obtain site-specific acid dissociation constants (pK A values) is challenging. In order to analyze the biphasic titrations exhibited by the side chain 13 C γ nuclei of the nucleophilic Glu78 and general acid/base Glu172 in Bacillus circulans xylanase, we have revisited the formalism for the ionization equilibria of two coupled acidic residues. In general, fitting NMR-monitored pH titration curves for such a system will only yield the two macroscopic pK A values that reflect the combined effects of both deprotonation reactions. However, through the use of mutations complemented with ionic strength-dependent measurements, we are able to extract the four microscopic pK Ai values governing the branched acid/base equilibria of Glu78 and Glu172 in BcX. These data, confirmed through theoretical calculations, help explain the pH-dependent mechanism of this model GH11 xylanase by demonstrating that the kinetically determined pK A values and hence catalytic roles of these two residues result from their electrostatic coupling.

  19. Influence of agitation speeds and aeration rates on the Xylanase activity of Aspergillus niger SS7

    Directory of Open Access Journals (Sweden)

    Yasser Bakri

    2011-08-01

    Full Text Available In this study, the effect of agitation and aeration rates on xylanase activity of Aspergillus niger SS7 in 3-litre stirred tank bioreactor was investigated. The agitation rates tested were 100, 200 and 300 rpm at each airflow rates of 0.5, 1.0 and 1.5 vvm. The maximum xylanase activity in mono- agitator system was at the agitation speed of 200 rpm and aeration rate of 1.0 vvm. In bi-agitator system, at low agitation speed (100 rpm, the xylanase activity was enhanced by 13% compared to mono- agitator system for an aeration rate of 1.0 vvm. Xylanase productivity in continuous culture was higher by approximately 3.5 times than in batch culture.

  20. GH10 xylanase D from Penicillium funiculosum: biochemical studies and xylooligosaccharide production

    Directory of Open Access Journals (Sweden)

    Giardina Thierry

    2011-04-01

    Full Text Available Abstract Background The filamentous fungus Penicillium funiculosum produces a range of glycoside hydrolases (GH. The XynD gene, encoding the sole P. funiculosum GH10 xylanase described so far, was cloned into the pPICZαA vector and expressed in methylotrophe yeast Pichia pastoris, in order to compare the results obtained with the P. funiculosum GH11 xylanases data. Results High level expression of recombinant XynD was obtained with a secretion of around 60 mg.L-1. The protein was purified to homogeneity using one purification step. The apparent size on SDS-PAGE was around 64 kDa and was 46 kDa by mass spectrometry thus higher than the expected molecular mass of 41 kDa. The recombinant protein was N- and O-glycosylated, as demonstrated using glycoprotein staining and deglycosylation reactions, which explained the discrepancy in molecular mass. Enzyme-catalysed hydrolysis of low viscosity arabinoxylan (LVAX was maximal at pH 5.0 with Km(app and kcat/Km(app of 3.7 ± 0.2 (mg.mL-1 and 132 (s-1mg-1.mL, respectively. The activity of XynD was optimal at 80°C and the recombinant enzyme has shown an interesting high thermal stability at 70°C for at least 180 min without loss of activity. The enzyme had an endo-mode of action on xylan forming mainly xylobiose and short-chain xylooligosaccharides (XOS. The initial rate data from the hydrolysis of short XOS indicated that the catalytic efficiency increased slightly with increasing their chain length with a small difference of the XynD catalytic efficiency against the different XOS. Conclusion Because of its attractive properties XynD might be considered for biotechnological applications. Moreover, XOS hydrolysis suggested that XynD possess four catalytic subsites with a high energy of interaction with the substrate and a fifth subsite with a small energy of interaction, according to the GH10 xylanase literature data.

  1. Modeling process for bioproduction of xylanase by Streptomyces spp. p12-137 on lignocelluloses agro-wastes

    Directory of Open Access Journals (Sweden)

    Gigi COMAN

    2012-12-01

    Full Text Available The production of xylanase without cellulase is required for the prebleaching of pulps, paper and food industry. The strain Streptomyces spp.P12-137 developed from the spores of the wild type organism was used in this work. Cultures in Erlenmeyer flasks, under shaking condition (150 rpm at temperature and pH values (28°C, 5.0 respectively revealed a xylanase activity of 27.77 IU·mL-1 after 120 h fermentation. This study demonstrates that Streptomyces spp. P12-137 is able to produce xylanase when wheat bran is used as a substrate. Fermentation was performed in a glass bioreactor withforced aeration. Data obtained have been compared to data from mathematical model obtained by numerical simulation using Matlab 7.9.0.529 (MathWorks, Inc. USA. The numerical simulation of the bioprocess could be a useful tool for adopting a control strategy to achieve increased xylanases yields under pilot or industrial conditions.

  2. Effect of pH on production of xylanase by Trichoderma reesei on xylan- and cellulose-based media

    Energy Technology Data Exchange (ETDEWEB)

    Bailey, M.J. (VTT, Biotechnical Lab., Espoo (Finland)); Buchert, J. (VTT, Biotechnical Lab., Espoo (Finland)); Viikari, L. (VTT, Biotechnical Lab., Espoo (Finland))

    1993-11-01

    Trichoderma reesei VTT-D-86271 (Rut C-30) was cultivated on media based on cellulose and xylan as the main carbon source in fermentors with different pH minimum controls. Production of xylanase was favoured by a rather high pH minimum control between 6.0 and 7.0 on both cellulose- and xylan-based media. Although xylanase was produced efficiently on cellulose as well as on xylan as the carbon source, significant production of cellulase was observed only on the cellulose-based medium and best production was at lower pH (4.0 minimum). Production of xylanase at pH 7.0 was shown to be dependent on the nature of the xylan in the cultivation medium but was independent of other organic components. Best production of xylanase was observed on insoluble, unsubstituted beech xylan at pH 7.0. Similar results were obtained in laboratory and pilot (200-1) fermentors. Downstream processing of the xylanase-rich, low-cellulase culture filtrate presented no technical problems despite apparent autolysis of the fungus at the high pH. Enzyme produced in the 200-1 pilot fermentor was shown to be suitable for use in enzyme-aided bleaching of kraft pulp. Due to the high xylanase/cellulase ratio of enzyme activities in the culture filtrate, pretreatment for removal of cellulase activity prior to pulp bleaching was unnecessary. (orig.)

  3. Novel structural features of xylanase A1 from Paenibacillus sp. JDR-2

    Science.gov (United States)

    Franz J. St John; James F. Preston; Edwin Pozharski

    2012-01-01

    The Gram-positive bacterium Paenibacillus sp. JDR-2 (PbJDR2) has been shown to have novel properties in the utilization of the abundant but chemically complex hemicellulosic sugar glucuronoxylan. Xylanase A1 of PbJDR2 (PbXynA1) has been implicated in an efficient process in which extracellular...

  4. Partial purification and characterization of xylanase produced from aspergillus niger using wheat bran

    International Nuclear Information System (INIS)

    Ahmad, Z.; Butt, M.S.

    2013-01-01

    In present exploration, purification and characterization of xylanase was carried out to find its optimum conditions for maximum functionality. The xylanase (EC 3.2.1.8) synthesized by Aspergillus niger in submerged fermentation was partially purified and characterized for different parameters like temperature, pH and heat stability. The molecular mass determined through SDS-PAGE was found 30 kDa. The specific activity of the enzyme was raised from 41.85 to 613.13 with 48.63% yield just in a two step partial purification comprising ammonium sulphate precipitation and Sephadex gel filteration column chromatography. The partially purified enzyme was found to be optimally active at 60 degree C and 7.5 pH. Conclusively, for the application of xylanase in food, feed or paper manufacturing processes, it is necessary to consider its optimum pH and temperature. (author)

  5. Production of xylanases by mangrove fungi from the Philippines and their application in enzymatic pretreatment of recycled paper pulps.

    Science.gov (United States)

    Torres, Jeremy Martin O; Dela Cruz, Thomas Edison E

    2013-04-01

    Mangrove fungi are vastly unexplored for enzymes with industrial application. This study aimed to assess the biocatalytic activity of mangrove fungal xylanases on recycled paper pulp. Forty-four mangrove fungal (MF) isolates were initially screened for xylanolytic activity in minimal medium with corn cob xylan as the sole carbon source. Eight MF were further cultivated under submerged fermentation for the production of crude xylanases. These crude enzymes were then characterized and tested for the pretreatment of recycled paper pulps. Results showed that 93 % of the tested MF isolates exhibited xylanolytic activity in solid medium. In submerged fermentation, salinity improved the growth of the fungal isolates but did not influence xylanase production. The crude xylanases were mostly optimally active at 50 °C and pH 7. Changes in pH had a greater effect on xylanase stability than temperature. More than half of the activity was lost at pH 9 for majority of the crude enzymes. However, two thermophilic xylanases from Fusarium sp. KAWIT-A and Aureobasidium sp. 2LIPA-M and one alkaliphilic xylanase from Phomopsis sp. MACA-J were also produced. All crude enzymes exhibited cellulase activities ranging from 4 to 21 U/ml. Enzymatic pretreatment of recycled paper pulps with 5 % consistency produced 70-650 mg of reducing sugars per gram of pulp at 50 °C after 60 min. The release of high amounts of reducing sugars showed the potential of mangrove fungal crude xylanases in the local paper and pulp industry. The diverse properties shown by the tested crude enzymes also indicate its potential applications to other enzyme-requiring industries.

  6. Effect of Cellulases and Xylanases on Refining Process and Kraft Pulp Properties.

    Directory of Open Access Journals (Sweden)

    Kamila Przybysz Buzała

    Full Text Available Samples of bleached kraft pine cellulosic pulp, either treated with an enzyme preparation (a Thermomyces lanuginosus xylanase, an Aspergillus sp. cellulase, and a multienzyme preparation NS-22086 containing both these activities or untreated, were refined in a laboratory PFI mill. The treatment with cellulases contained in the last two preparations significantly improved the pulp's susceptibility to refining (the target freeness value of 30°SR was achieved in a significantly shorter time, increased water retention value (WRV and fines contents while the weighted average fiber length was significantly reduced. These changes of pulp parameters caused deterioration of paper strength properties. The treatment with the xylanase, which partially hydrolyzed xylan, small amounts of which are associated with cellulose fibers, only slightly loosened the structure of fibers. These subtle changes positively affected the susceptibility of the pulp to refining (refining energy was significantly reduced and improved the static strength properties of paper. Thus, the treatment of kraft pulps with xylanases may lead to substantial savings of refining energy without negative effects on paper characteristics.

  7. Thermal-induced conformational changes in the product release area drive the enzymatic activity of xylanases 10B: Crystal structure, conformational stability and functional characterization of the xylanase 10B from Thermotoga petrophila RKU-1

    International Nuclear Information System (INIS)

    Santos, Camila Ramos; Meza, Andreia Navarro; Hoffmam, Zaira Bruna; Silva, Junio Cota; Alvarez, Thabata Maria; Ruller, Roberto; Giesel, Guilherme Menegon; Verli, Hugo; Squina, Fabio Marcio; Prade, Rolf Alexander; Murakami, Mario Tyago

    2010-01-01

    Research highlights: → The hyperthermostable xylanase 10B from Thermotoga petrophila RKU-1 produces exclusively xylobiose at the optimum temperature. → Circular dichroism spectroscopy suggests a coupling effect of temperature-induced structural changes with its enzymatic behavior. → Crystallographic and molecular dynamics studies indicate that conformational changes in the product release area modulate the enzyme action mode. -- Abstract: Endo-xylanases play a key role in the depolymerization of xylan and recently, they have attracted much attention owing to their potential applications on biofuels and paper industries. In this work, we have investigated the molecular basis for the action mode of xylanases 10B at high temperatures using biochemical, biophysical and crystallographic methods. The crystal structure of xylanase 10B from hyperthermophilic bacterium Thermotoga petrophila RKU-1 (TpXyl10B) has been solved in the native state and in complex with xylobiose. The complex crystal structure showed a classical binding mode shared among other xylanases, which encompasses the -1 and -2 subsites. Interestingly, TpXyl10B displayed a temperature-dependent action mode producing xylobiose and xylotriose at 20 o C, and exclusively xylobiose at 90 o C as assessed by capillary zone electrophoresis. Moreover, circular dichroism spectroscopy suggested a coupling effect of temperature-induced structural changes with this particular enzymatic behavior. Molecular dynamics simulations supported the CD analysis suggesting that an open conformational state adopted by the catalytic loop (Trp297-Lys326) provokes significant modifications in the product release area (+1,+2 and +3 subsites), which drives the enzymatic activity to the specific release of xylobiose at high temperatures.

  8. Thermal-induced conformational changes in the product release area drive the enzymatic activity of xylanases 10B: Crystal structure, conformational stability and functional characterization of the xylanase 10B from Thermotoga petrophila RKU-1

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Camila Ramos; Meza, Andreia Navarro [Laboratorio Nacional de Biociencias (LNBio), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Hoffmam, Zaira Bruna; Silva, Junio Cota; Alvarez, Thabata Maria; Ruller, Roberto [Laboratorio Nacional de Ciencia e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Giesel, Guilherme Menegon; Verli, Hugo [Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS (Brazil); Squina, Fabio Marcio [Laboratorio Nacional de Ciencia e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Prade, Rolf Alexander [Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK (United States); Murakami, Mario Tyago, E-mail: mario.murakami@lnbio.org.br [Laboratorio Nacional de Biociencias (LNBio), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil)

    2010-12-10

    Research highlights: {yields} The hyperthermostable xylanase 10B from Thermotoga petrophila RKU-1 produces exclusively xylobiose at the optimum temperature. {yields} Circular dichroism spectroscopy suggests a coupling effect of temperature-induced structural changes with its enzymatic behavior. {yields} Crystallographic and molecular dynamics studies indicate that conformational changes in the product release area modulate the enzyme action mode. -- Abstract: Endo-xylanases play a key role in the depolymerization of xylan and recently, they have attracted much attention owing to their potential applications on biofuels and paper industries. In this work, we have investigated the molecular basis for the action mode of xylanases 10B at high temperatures using biochemical, biophysical and crystallographic methods. The crystal structure of xylanase 10B from hyperthermophilic bacterium Thermotoga petrophila RKU-1 (TpXyl10B) has been solved in the native state and in complex with xylobiose. The complex crystal structure showed a classical binding mode shared among other xylanases, which encompasses the -1 and -2 subsites. Interestingly, TpXyl10B displayed a temperature-dependent action mode producing xylobiose and xylotriose at 20 {sup o}C, and exclusively xylobiose at 90 {sup o}C as assessed by capillary zone electrophoresis. Moreover, circular dichroism spectroscopy suggested a coupling effect of temperature-induced structural changes with this particular enzymatic behavior. Molecular dynamics simulations supported the CD analysis suggesting that an open conformational state adopted by the catalytic loop (Trp297-Lys326) provokes significant modifications in the product release area (+1,+2 and +3 subsites), which drives the enzymatic activity to the specific release of xylobiose at high temperatures.

  9. Polypeptides having xylanase activity and polynucleotides encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Spodsberg, Nikolaj

    2018-02-06

    The present invention relates to isolated polypeptides having xylanase activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

  10. Preparation of arabinoxylobiose from rye xylan using family 10 Aspergillus aculeatus endo-1,4-ß-d-xylanase

    NARCIS (Netherlands)

    Rantanen, H.; Virkki, L.; Tuomainen, P.; Kabel, M.A.; Schols, H.A.; Tenkanen, M.

    2007-01-01

    Commercial xylanase preparation Shearzyme®, which contains the glycoside hydrolase family 10 endo-1,4-ß-d-xylanase from Aspergillus aculeatus, was used to prepare short-chain arabinoxylo-oligosaccharides (AXOS) from rye arabinoxylan (AX). A major AXOS was formed as a hydrolysis product. Longer AXOS

  11. Influence of a direct-fed microbial and xylanase enzyme on the dietary energy uptake efficiency and performance of broiler chickens.

    Science.gov (United States)

    Murugesan, Ganapathi Raj; Persia, Michael E

    2015-09-01

    Efficacy of a multi-strain direct-fed microbial product (PoultryStar(®) ME; PS) and a xylanase enzyme product on the dietary energy utilization efficiency and resulting performance in broiler chickens was evaluated. Apart from performance parameters, cecal and serum metabolites and activities of hepatic enzymes involved in energy metabolism were also determined. Ross 308 chicks were fed one of four experimental diets [control (CON), CON + PS, CON + xylanase and CON + PS + xylanase] using a 2 × 2 factorial arrangement from 1-21 days of age. Cecal proportions of propionate and butyrate, as well as total short-chain fatty acid concentration were increased (P energy uptake and hepatic energy retention. The combination additively increased the FCR, suggesting involvement of synergistic modes of actions. © 2014 Society of Chemical Industry.

  12. Bifunctional xylanases and their potential use in biotechnology

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Numan, M.Th.

    . J Chromatography 919:389–394 33. Hong SY, Lee JS, Cho KM, Math RK, Kim YH, Hong SJ, Cho YU, Kim H, Yun HD (2006) Assembling a novel bifunctional cel- lulase–xylanase from Thermotoga maritima by end-to-end fusion. Biotechnol Lett 28:1857–1862 34...

  13. Screening of Thermophilic Bacteria Produce Xylanase from Sapan Sungai Aro Hot Spring South Solok

    Science.gov (United States)

    Irdawati, I.; Syamsuardi, S.; Agustien, A.; Rilda, Y.

    2018-04-01

    xylanase is one of the enzymes with great prospects as hemicellulose hydrolyzing enzyme. Global annual market demand for this enzyme reach US 200 million. This enzyme catalyzes the xylan (hemicellulose) reactions breaking into xilooligosakarida and xylose. Xylanase can be applied to various industrial sectors such as bread, sugar xylose, biofuels, especially in bleaching paper (bleaching) pulp. Xylanase Isable to replace conventional chemical bleaching using chlorine that is not friendly for the environment. Currently xylanase production is extracted from the thermophilic bacteria for enzyme stability at high temperatures that are suitable for industrial applications. Thermophilic bacteria can be isolated from a hot spring, one of the which is a source of Sapan Sungai Aro Hot Spring, located in the district South Solok. The aim of this study was to select and identification of thermophilic bacteria can produce xylanase.This roomates is a descriptive study, which was Carried out in the Laboratory of Microbiology, Mathematic and Science Faculty of Padang State University, and Laboratory of Bacteriology, BasoVeterinary Research Center. The research procedure consisted of the preparation and sterilization of materials and tools, medium manufacturing, regeneration, selection and identification. Selection is performed by using a semiquantitative screening plate that contains xylan substrate. Identification is based on microscopic and biochemical characteristics until the genus level.Selection results Showed 12 out of 16 isolates had xilanolitik activity, with the highest activity is SSA2 with xilanolitik index of 0.74. The top five index producehigestxilanolitik isolates that are SSA2, SSA3 and SSA4 identified as Bacillus sp. 1., and SSAS6 and SSA7 is Bacillus sp. 2.

  14. Screening of culture condition for xylanase production by ...

    African Journals Online (AJOL)

    The study demonstrated not only the importance of the nature of the substrate in obtaining a system resistant to catabolic repression, but also the importance of the culture conditions for biosynthesis of this enzyme. T. viride showed a high potential for xylanase production under the conditions presented in these assays.

  15. High xylanase production by Trichoderma viride using pineapple ...

    African Journals Online (AJOL)

    Xylanases are hydrolases which depolymerize the xylan components present in plants cell wall. Commercial applications for these enzymes include its use in the pulp bleaching, food and animal feed industries, among others. Recently, there is a great interest on the exploitation of agro-industrial wastes as low-cost raw ...

  16. Variation in levels of non-starch polysaccharides and endogenous endo-1,4-β-xylanases affects the nutritive value of wheat for poultry.

    Science.gov (United States)

    Cardoso, V; Fernandes, E A; Santos, H M M; Maçãs, B; Lordelo, M M; Telo da Gama, Luis; Ferreira, L M A; Fontes, C M G A; Ribeiro, T

    2018-04-01

    1. Endo-1,4-β-xylanase is known to improve the nutritive value of wheat-based diets for poultry by degrading dietary arabinoxylans. However, broilers' response to supplementation of wheat-based diets with exogenous endo-1,4-β-xylanase is not always observed. 2. In this study, 108 different wheat lots were analysed for levels of extract viscosity as well as for endogenous endo-1,4-β-xylanase activity, and the impact of these two variables in animal performance was tested. 3. Results revealed that endogenous endo-1,4-β-xylanase activity and extract viscosity content varied widely among different wheat lots. Thus, a trial was conducted to evaluate the efficacy of exogenous enzyme supplementation in broiler diets using wheats with different levels of extract viscosity and endogenous endo-1,4-β-xylanase activity. 4. The data revealed that exogenous enzyme supplementation was only effective when the wheat present in the diet had high levels of extract viscosity (14.8 cP) with low endogenous endo-1,4-β-xylanase activity (347.0 U/kg). Nevertheless, it is apparent that exogenous microbial xylanases reduce digesta extract viscosity and feed conversion ratio independently of the endogenous properties presented by different wheat lots. 5. The data suggest that extract viscosity and/or endogenous endo-1,4-β-xylanase activity affect the response to enzyme supplementation by poultry fed on wheat-based diets.

  17. Enhanced production of xylanase from locally isolated fungal strain using agro-industrial residues under solid-state fermentation.

    Science.gov (United States)

    Abdullah, Roheena; Nisar, Kinza; Aslam, Aafia; Iqtedar, Mehwish; Naz, Shagufta

    2015-01-01

    This study is related to the isolation of fungal strain for xylanase production using agro-industrial residues. Forty fungal strains with xylanolytic potential were isolated by using xylan agar plates and quantitatively screened in solid-state fermentation. Of all the tested isolates, the strain showing highest ability to produce xylanase was assigned the code Aspergillus niger LCBT-14. For the enhanced production of the enzyme, five different fermentation media were evaluated. Out of all media, M4 containing wheat bran gave maximum enzyme production. Effect of different variables including incubation time, temperature, pH, carbon and nitrogen sources has been investigated. The optimum enzyme production was obtained after 72 h at 30°C and pH 4. Glucose as a carbon source while ammonium sulphate and yeast extract as nitrogen sources gave maximum xylanase production (946 U/mL/min). This study was successful in producing xylanase by A. niger LCBT-14 economically by utilising cheap indigenous substrate.

  18. Disruption of the L-arabitol dehydrogenase encoding gene in Aspergillus tubingensis results in increased xylanase production

    NARCIS (Netherlands)

    Nikolaev, I.; Hansen, S.; Madrid, S.; de Vries, R.P.

    2013-01-01

    Fungal xylanases are of major importance to many industrial sectors, such as food and feed, paper and pulp, and biofuels. Improving their production is therefore highly relevant. We determined the molecular basis of an improved xylanase-producing strain of Aspergillus tubingensis that was generated

  19. Acidic-alkaline ferulic acid esterase from Chaetomium thermophilum var. dissitum: Molecular cloning and characterization of recombinant enzyme expressed in Pichia pastoris

    DEFF Research Database (Denmark)

    Dotsenko, Gleb; Tong, Xiaoxue; Pilgaard, Bo

    2016-01-01

    A novel ferulic acid esterase encoding gene CtFae, was successfully cloned from a highly esterase active strain of the thermophile ascomycetous fungus Chaetomium thermophilum var. dissitum; the gene was heterologously expressed in Pichia pastoris KM71H. The recombinant enzyme (CtFae) was purified...... to homogeneity and subsequently characterized. CtFae was active towards synthetic esters of ferulic, p-coumaric, and caffeic acids, as well as towards wide range of p-nitrophenyl substrates. Its temperature and pH optima were 55 °C and pH 6.0, respectively. Enzyme rare features were broad pH optimum, high...

  20. Optimization of xylanase production by Mucor indicus, Mucor hiemalis, and Rhizopus oryzae through solid state fermentation

    Directory of Open Access Journals (Sweden)

    Sanaz Behnam

    2016-03-01

    Full Text Available Introduction: Xylan is the main hemicellulosic polymer in a number of lignocelluloses which can be hydrolyzed by xylanolytic enzymes. One of the main ways for enzymes production is solid state fermentation (SSF. The ability of three fungal strains (Mucor indicus, Mucor hiemalis, and Rhizopus oryzae for xylanase production on wheat bran by SSF was investigated. Materials and methods: The effects of cultivation temperature, medium moisture content, and cultivation time on the enzyme production were investigated. Experiments were designed with an orthogonal central composite design on three variables using response surface methodology (RSM. Analysis of variance was applied and the enzyme production was expressed with a mathematical equation as a function of the three factors. The optimum operating conditions for the enzyme production was obtained. Results: For xylanase production by M. indicus, M. hiemalis and R. oryzae the optimum temperatures were 40.0, 43.4 and 43.4ºC respectively. These values were 49.8, 54.2 and 71.8% for moisture percent and 51.3, 53.2 and 53.5 h for cultivation time. The highest enzyme activities per g of dry substrate (gds were 43.1, 43.8 and 25.9 U/gds for M. indicus, M. hiemalis and R. oryzae respectively. Discussion and conclusion: All the fungi were able to produce xylanase. Maximum xylanase production was predicted by M. indicus and M. hiemalis at similar optimum conditions, while R. oryzae produced relatively lower xylanase activity even at the best condition. 

  1. Paddy Husk as Support for Solid State Fermentation to Produce Xylanase from Bacillus pumilus

    Directory of Open Access Journals (Sweden)

    Ranganathan KAPILAN

    2011-03-01

    Full Text Available To optimize culture conditions for xylanase production by solid state fermentation (SSF using Bacillus pumilus, with paddy husk as support, solid medium contained 200 g of paddy husk with 800 mL of liquid fermentation medium [xylan, 20.0 g/L; peptone, 2.0 g/L; yeast extract, 2.5 g/L; K2HPO4, 2.5 g/L; KH2PO4, 1.0 g/L; NaCl, 0.1 g/L; (NH42SO4, 2.0 g/L, CaCl2·2H2O, 0.005 g/L; MgCl2·6H2O, 0.005 g/L; and FeCl3, 0.005 g/L] at pH 9.0 was applied. The highest xylanase activity (142.0 ±0.47 U/g DM] was obtained on the 6th day at 30°C. The optimized paddy husk to liquid fermentation medium ratio was 2:9, and the optimized culture temperature was 40°C. When commercial Birchwood xylan was replaced with different concentrations of corncob, xylanase production was maximized (224.2 U/g DM in the medium with 150 g/L corncob. Xylanase production was increased by sucrose, fructose and arabinose, whereas reduced by glucose, galactose, lactose and amylose. When organic nitrogen sources were replaced with locally available nitrogen sources such as groundnut powder or sesame seedcake powder or coconut seedcake powder or soy meal powder, the highest xylanase production (290.7 U/g DM was obtained in the medium with soy meal powder and 16.0 g/L of soy meal powder was the optimum (326.5±0.34 U/g DM. Based on the optimization studies, B. pumilus produced 2.3 times higher xylanase activity. The medium cost was reduced from 2 458.3 to 178.3 SLR/kg and the total activity which could be obtained from 1 kg of the medium was increased from 48 624 to 220 253 Units.

  2. Production of Xylanase by Recombinant Bacillus subtilis DB104 Cultivated in Agroindustrial Waste Medium

    Directory of Open Access Journals (Sweden)

    Is Helianti

    2016-07-01

    Full Text Available A recombinant Bacillus subtilis DB104 strain harbouring recombinant plasmid pSKE194 containing an Open Reading Frame (ORF of endoxylanase and its indigenous promoter from the wild-type B. subtilis AQ1 strain was constructed. This recombinant B. subtilis DB104 strain had higher endoxylanase activity than the nonrecombinant B. subtilis DB104 strain in standard media, such as Luria Bertani (LB and LB with xylan. The agroindustrial wastes corncobs and tofu liquid waste were chosen as cost-effective carbon and nitrogen sources, respectively, to test the economics of xylanase production using the recombinant B. subtilis DB104 at a larger scale. Submerged fermentation using a 4.5 L working volume fermentor with tofu liquid waste and 4% corncobs produced maximum xylanase activity of 1296 ± 1.2 U/mg (601.7 ± 0.6 U/mL after 48-hour fermentation at 37°C with 150 rpm agitation; this is more than twofold higher than the activity produced in an Erlenmeyer flask. This is the first report of high xylanase activity produced from recombinant B. subtilis using inexpensive medium. During fermentation, the xylanase degrades corncobs into xylooligosaccharides, showing its potential as an enzyme feed additive or in xylooligosaccharide production.

  3. Cloning, sequencing and expression of a xylanase gene from the maize pathogen Helminthosporium turcicum

    DEFF Research Database (Denmark)

    Degefu, Y.; Paulin, L.; Lübeck, Peter Stephensen

    2001-01-01

    A gene encoding an endoxylanase from the phytopathogenic fungus Helminthosporium turcicum Pass. was cloned and sequenced. The entire nucleotide sequence of a 1991 bp genomic fragment containing an endoxylanase gene was determined. The xylanase gene of 795 bp, interrupted by two introns of 52 and ...

  4. Molecular Dynamics Approach in Designing Thermostable Aspergillus niger Xylanase

    Science.gov (United States)

    Malau, N. D.; Sianturi, M.

    2017-03-01

    Molecular dynamics methods we have applied as a tool in designing thermostable Aspergillus niger Xylanase, by examining Root Mean Square Deviation (RMSD) and The Stability of the Secondary Structure of enzymes structure at its optimum temperature and compare with its high temperature behavior. As RMSD represents structural fluctuation at a particular temperature, a better understanding of this factor will suggest approaches to bioengineer these enzymes to enhance their thermostability. In this work molecular dynamic simulations of Aspergillus niger xylanase (ANX) have been carried at 400K (optimum catalytic temperature) for 2.5 ns and 500K (ANX reported inactive temperature) for 2.5 ns. Analysis have shown that the Root Mean Square Deviation (RMSD) significant increase at higher temperatures compared at optimum temperature and some of the secondary structures of ANX that have been damaged at high temperature. Structural analysis revealed that the fluctuations of the α-helix and β-sheet regions are larger at higher temperatures compared to the fluctuations at optimum temperature.

  5. Evaluation of the effect of different wheats and xylanase supplementation on performance, nutrient and energy utilisation in broiler chicks

    Directory of Open Access Journals (Sweden)

    Gemma González-Ortiz

    2016-09-01

    Full Text Available The aim of this study was to evaluate the performance, nutrient utilisation and energy metabolism of broiler chicks fed 8 different wheat samples, supplemented or not with xylanase. Seven-hundred sixty eight male broilers (1-day-old were distributed to 16 experimental treatments (6 replicates per treatment. The treatments were in a factorial arrangement with 8 different wheats and 2 levels of xylanase (0 or 16,000 BXU/kg. The predicted apparent metabolisable energy (AME of the wheat samples ranged from 13.0 to 13.9 MJ/kg and all diets were formulated to contain the same amount of wheat. Body weight gain (BWG and feed intake (FI were measured at 21 d, as was jejunal digesta viscosity, and feed conversion ratio (FCR calculated. On day 24, one representative bird per pen was selected to calculate whole body energetics. At 21 d, 3 chicks per replicate were randomly allocated to metabolism cages for energy and nutrient utilisation determinations, and were continued on the experimental diets until 24-d-old. No interactions were observed for any performance response variables, ileal nutrient utilisation or digesta viscosity. Xylanase improved BWG and reduced FCR and digesta viscosity (P < 0.05. Wheat influenced dry matter (DM utilisation and xylanase increased ileal digestible energy (P = 0.04. Xylanase also improved (P < 0.05 DM and nitrogen retention. Apparent metabolisable energy and AME corrected for nitrogen (AMEn were subject to an interaction whereby wheats 2 and 6, which returned the lowest AME and AMEn values, responded to xylanase supplementation and the remainder did not. Net energy for production and the efficiency of energy use for production were not influenced by xylanase, but were affected by wheat (P < 0.05. Despite the significant differences between wheats with regards to their nutrient utilisation and energy metabolism in birds, xylanase removed this variance and resulted in more homogeneous performance.

  6. Enzymatic saccharification of seaweeds into fermentable sugars by xylanase from marine Bacillus sp. strain BT21.

    Science.gov (United States)

    Parab, Pankaj; Khandeparker, Rakhee; Amberkar, Ujwala; Khodse, Vishwas

    2017-10-01

    Enzymatic hydrolysis of seaweed biomass was studied using xylanase produced from marine bacteria Bacillus sp. strain BT21 through solid-state fermentation of wheat bran. Three types of seaweeds, Ahnfeltia plicata , Padina tetrastromatica and Ulva lactuca , were selected as representatives of red, brown, and green seaweeds, respectively. Seaweed biomass was pretreated with hot water. The efficiency of pretreated biomass to release reducing sugar by the action of xylanase as well as the type of monosaccharide released during enzyme saccharification of seaweed biomass was studied. It was seen that pretreated biomass of seaweed A. plicata, U. lactuca , and P. tetrastroma , at 121 °C for 45 min, followed by incubation with 50 IU xylanase released reducing sugars of 233 ± 5.3, 100 ± 6.1 and 73.3 ± 4.1 µg/mg of seaweed biomass, respectively. Gas chromatography analysis illustrated the release of xylose, glucose, and mannose during the treatment process. Hot water pre-treatment process enhanced enzymatic conversion of biomass into sugars. This study revealed the important role of xylanase in saccharification of seaweed, a promising feedstock for third-generation bioethanol production.

  7. Optimization of moistening solution concentration on xylanase activity in solid state fermentation from oil palm empty fruit bunches

    Science.gov (United States)

    Mardawati, Efri; Parlan; Rialita, Tita; Nurhadi, Bambang

    2018-03-01

    Xylanase is an enzyme used in the industrial world, including food industry. Xylanase can be utilized as a 1,4-β-xylosidic endo-hydrolysis catalyst of xylanase, a hemicellulose component for obtaining a xylose monomer. This study aims to determine the optimum concentration of the fermentation medium using Response Surface Method (RSM) in the production of xylanase enzyme from oil palm empty fruit bunches (OPEFB) through solid state fermentation process. The variables varied in this study used factor A (ammonium sulphate concentration 1.0-2.0 g/L), B (concentration of potassium dihydrogen phosphate 1.5-2.5 g/L) and C (urea concentration 0.2 – 0.5 g/L). The data was analysed by using Design Expert version 10.0.1.0 especially CCD with total 17 running including 3 times replicated of canter point. Trichoderma viride was used for the process production of xylanase enzyme. The ratio between substrate and moistening solution used was 0.63 g / mL with temperature of 32.80C, 60 h incubation time. The analysis of enzyme activity was done by DNS method with 1% xylan as substrate. Analysis of protein content in enzyme was done by Bradford method. The optimum of moistening solution concentration in this fermentation was obtained. They are, the ammonium sulphate concentration of 1.5 g/L, potassium dihydrogen phosphate 2.0 g/L and urea 0.35 g/L with activity of 684.70 U/mL, specific activity enzyme xylanase 6261.58 U/mg, protein content 0.1093 U/mg, the model was validated using experiment design with perfect reliability value 0.96.

  8. (Trametes sp.) in the production of cellulase and xylanase

    African Journals Online (AJOL)

    Nanda

    2016-05-18

    May 18, 2016 ... in solid-sate fermentation (SSF), in this work, the production of cellulase and xylanase by the fungus ... sugars can be converted to ethanol, lactic acid and ... substances; clarification of juices and wines; improving ..... SSF processes has a marked effect on growth kinetics, ..... Overview of applied solid-state.

  9. Potential of Laceyella sacchari strain B42 crude xylanase in ...

    African Journals Online (AJOL)

    rajmac

    2013-02-06

    Feb 6, 2013 ... sacchari strain B42. Maximal xylanase production was achieved at the incubation period of 48 h with ... chemicals for pulp processing and bleaching. The major ... industrial enzyme with great biotechnological application.

  10. Ultrasounds pretreatment of olive pomace to improve xylanase and cellulase production by solid-state fermentation.

    Science.gov (United States)

    Leite, Paulina; Salgado, José Manuel; Venâncio, Armando; Domínguez, José Manuel; Belo, Isabel

    2016-08-01

    Olive mills generate a large amount of waste that can be revaluated. This work aim to improve the production lignocellulolytic enzymes by solid-state fermentation using ultrasounds pretreated olive mill wastes. The composition of olive mill wastes (crude and exhausted olive pomace) was compared and several physicochemical characteristics were significantly different. The use of both wastes in SSF was evaluated and a screening of fungi for xylanase and cellulase production was carried out. After screening, the use of exhausted olive pomace and Aspergillus niger led to the highest enzyme activities, so that they were used in the study of ultrasounds pre-treatment. The results showed that the sonication led to a 3-fold increase of xylanase activity and a decrease of cellulase activity. Moreover, the liquid fraction obtained from ultrasounds treatment was used to adjust the moisture of solid and a positive effect on xylanase (3.6-fold increase) and cellulase (1.2-fold increase) production was obtained. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Xylanase from Fusarium heterosporum: Properties and influence of ...

    African Journals Online (AJOL)

    Unioeste

    2014-02-26

    Feb 26, 2014 ... influence of thiol compounds on xylanase activity. Paulo Ricardo ... and the extraction of plant oil, coffee and starch (Ahmed ... of F. heterosporum on 5 g of various carbon sources ... brewing residue under solid-state fermentation (fungal strain and .... active within the acidic pH range of 4.5 to 5.5 and.

  12. Comparison of kinetic characteristics of xylanases from Aspergillus ...

    African Journals Online (AJOL)

    Arabinoxylans are the predominant non-starch polysaccharides of the cell walls of wheat grain, and can contribute up to 3% of the total polysaccharide content of the flour. Endo-(1-4)-β-xylanase is able to hydrolyze the glycosidic bonds between two xylose units in the xylan backbone during baking process. The use of ...

  13. High xylanase production by Trichoderma viride using pineapple ...

    African Journals Online (AJOL)

    SAM

    2014-05-28

    May 28, 2014 ... T. viride xylanase was stable at 40°C, showing the half-life (T1/2) value of ... Many studies have demonstrated that the pulp ... disposed in the environment without an adequate .... one-way analysis of variance and compared through the Tukey test, .... production by T. harzianum with wheat straw increases.

  14. Polypeptides having xylanase activity and polynucleotides encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Spodsberg, Nikolaj; Shaghasi, Tarana

    2017-06-20

    The present invention relates to polypeptides having xylanase activity, catalytic domains, and carbohydrate binding domains, and polynucleotides encoding the polypeptides, catalytic domains, and carbohydrate binding domains. The present invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides, catalytic domains, and carbohydrate binding domains.

  15. Evaluation of xylanases from Aspergillus niger and Trichoderma sp ...

    African Journals Online (AJOL)

    Despite being present in relatively low amounts, pentosans and hemicelluloses play an important role in dough rheology and bread properties. The aim of this work is to understand how the xylanases from Aspergillus niger and Trichoderma sp. influence dough rheology, such as elasticity, extensibility, strength and stability.

  16. Optimisation of amylase and xylanase addition in dependance of white flour amylase activity

    Directory of Open Access Journals (Sweden)

    Lončar Davor M.

    2016-01-01

    Full Text Available In this study the effect of different quantities of added amylase to white wheat flours characterized with different activities of naturally existing amylases is tested. Response surface methodology is chosen to test the effects of main applied technological parameters on bread quality responses. Independent variables are chosen to be: quantity of added amylase and bulk fermentation time, while analysed responses are: specific volume, grain structure, bulk fermentation. Bread quality responses are statistically significant, while predicted and observed responses correspond very well, which allows good prediction of bread quality parameters based on applied technological parameters and flour characteristics. Score analysis shows that optimum quantity of amylase addition regarding bread quality depends on the activity of naturally existing amylases. Optimal quantity of added xylanase in bread samples made from both flour types is 0.004%. Xylanase improved properties of white wheat bread and higher effect is experienced with flour that has more active naturally existing amylases. Addition of amylase has statistically significantly increased a* values of crust. Addition of xylanase has statistically significantly decreased values of b* in comparison to the respective bread sample with only added amylase.

  17. One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash

    Directory of Open Access Journals (Sweden)

    Ashwani Sanghi

    2010-06-01

    Full Text Available The present study describes the one-step purification and characterization of an extracellular cellulase-free xylanase from a newly isolated alkalophilic and moderately thermophilic strain of Bacillus subtilis ASH. Xylanase was purified to homogeneity by 10.5-fold with ~43% recovery using ion-exchange chromatography through CM-Sephadex C-50. The purified enzyme revealed a single band on SDS-PAGE gel with a molecular mass of 23 kDa. It showed an optimum pH at 7.0 and was stable over the pH range 6.0-9.0. The optimum temperature for enzyme activity was 55 ºC. The purified xylanase did not lose any activity up to 45 ºC, however, it retained 80% and 51% of its activity after pre-incubation at 55 ºC and 60 ºC, respectively. The enzyme obeyed Michaelis-Menton kinetics towards birch wood xylan with apparent Km 3.33 mg/ml and Vmax 100 IU/ml. The enzyme was strongly inhibited by Hg2+ and Cu2+ while enhanced by Co2+ and Mn2+. The purified enzyme could be stored at 4 ºC for six weeks without any loss of catalytic activity. The faster and economical purification of the cellulase-free xylanase from B. subtilis ASH by one-step procedure together with its appreciable stability at high temperature and alkaline pH makes it potentially effective for industrial applications.

  18. Constitutive expression of the xylanase inhibitor TAXI-III delays Fusarium head blight symptoms in durum wheat transgenic plants.

    Science.gov (United States)

    Moscetti, Ilaria; Tundo, Silvio; Janni, Michela; Sella, Luca; Gazzetti, Katia; Tauzin, Alexandra; Giardina, Thierry; Masci, Stefania; Favaron, Francesco; D'Ovidio, Renato

    2013-12-01

    Cereals contain xylanase inhibitor (XI) proteins which inhibit microbial xylanases and are considered part of the defense mechanisms to counteract microbial pathogens. Nevertheless, in planta evidence for this role has not been reported yet. Therefore, we produced a number of transgenic plants constitutively overexpressing TAXI-III, a member of the TAXI type XI that is induced by pathogen infection. Results showed that TAXI-III endows the transgenic wheat with new inhibition capacities. We also showed that TAXI-III is correctly secreted into the apoplast and possesses the expected inhibition parameters against microbial xylanases. The new inhibition properties of the transgenic plants correlate with a significant delay of Fusarium head blight disease symptoms caused by Fusarium graminearum but do not significantly influence leaf spot symptoms caused by Bipolaris sorokiniana. We showed that this contrasting result can be due to the different capacity of TAXI-III to inhibit the xylanase activity of these two fungal pathogens. These results provide, for the first time, clear evidence in planta that XI are involved in plant defense against fungal pathogens and show the potential to manipulate TAXI-III accumulation to improve wheat resistance against F. graminearum.

  19. Improvement for enhanced xylanase production by Cellulosimicrobium cellulans CKMX1 using central composite design of response surface methodology.

    Science.gov (United States)

    Walia, Abhishek; Mehta, Preeti; Guleria, Shiwani; Shirkot, Chand Karan

    2015-12-01

    The effects of yeast extract (X 1 ), NH 4 NO 3 (X 2 ), peptone (X 3 ), urea (X 4 ), CMC (X 5 ), Tween 20 (X 6 ), MgSO 4 (X 7 ), and CaCO 3 (X 8 ) on production of xylanase from Cellulosimicrobium cellulans CKMX1 were optimized by statistical analysis using response surface methodology (RSM). The RSM was used to optimize xylanase production by implementing the Central composite design. Statistical analysis of the results showed that the linear, interaction and quadric terms of these variables had significant effects. However, only the linear effect of X 4 , X 5 , interaction effect of X 1 X 7 , X 1 X 8 , X 2 X 3 , X 2 X 8 , X 3 X 6 , X 3 X 8 , X 4 X 6 , X 4 X 7 , X 5 X 7 , X 5 X 8 and quadratic effect of X 3 2 , X 5 2 and X 7 2 found to be insignificant terms in the quadratic model and had no response at significant level. The minimum and maximum xylanase production obtained was 331.50 U/g DBP and 1027.65 U/g DBP, respectively. The highest xylanase activity was obtained from Run No. 30, which consisted of yeast extract (X 1 ), 1.00 g (%); NH 4 NO 3 (X 2 ), 0.20 g (%); peptone (X 3 ), 1.00 g (%); urea (X 4 ), 10 mg (%); CMC (X 5 ), 1.00 g (%); Tween 20 (X 6 ), 0.02 mL (%); CaCO 3 (X 7 ), 0.50 g (%) and MgSO 4 (X 8 ), 9.0 g (%). The optimization resulted in 3.1-fold increase of xylanase production, compared with the lowest xylanase production of 331.50 U/g DBP after 72 h of incubation in stationary flask experiment. Application of cellulase-free xylanase in pulp biobleaching from C. cellulans CKMX1 under C-E P -D sequence has been shown to bring about a 12.5 % reduction of chlorine, decrease of 0.8 kappa points (40 %), and gain in brightness was 1.42 % ISO points in 0.5 % enzyme treated pulp as compared to control.

  20. Comparison of several ethanol productions using xylanase, inorganic salts, surfactant

    Science.gov (United States)

    Wu, Yan; Lu, Jie; Yang, Rui-feng; Song, Wen-jing; Li, Hai-ming; Wang, Hai-song; Zhou, Jing-hui

    2017-03-01

    Liquid hot water (LHW) pretreatment is an effective and environmentally friendly method to produce bioethanol with lignocellulosic materials. Corn stover was pretreated with liquid hot water (LHW) and then subjected to semi-simultaneous saccharification and fermentation (S-SSF) to obtain high ethanol concentration and yield. The present study aimed to confirm the effect of several additives on the fermentation digestibility of unwashed WIS of corn stover pretreated with LHW. So we also investigated the process, such as enzyme addition, inorganic salts, surfactant and different loading Triton. Results show that high ethanol concentration is necessary to add xylanase in the stage of saccharification. The ethanol concentration increased mainly with magnesium ion on fermentation. Comparing with Tween 80, Span 80 and Polyethylene glycol, Triton is the best surfactant. In contrast to using xylanase and Triton respectively, optimization can make up the lack of stamina and improve effect of single inorganic salts.

  1. Continuous xylanase production with Aspergillus nidulans under pyridoxine limitation using a trickle bed reactor.

    Science.gov (United States)

    Müller, Michael; Prade, Rolf A; Segato, Fernando; Atiyeh, Hasan K; Wilkins, Mark R

    2015-01-01

    A trickle bed reactor (TBR) with recycle was designed and tested using Aspergillus nidulans with a pyridoxine marker and over-expressing/secreting recombinant client xylanase B (XynB). The pyridoxine marker prevented the fungus from synthesizing its own pyridoxine and fungus was unable to grow when no pyridoxine was present in the medium; however, enzyme production was unaffected. Uncontrolled mycelia growth that led to clogging of the TBR was observed when fungus without a pyridoxine marker was used for XynB production. Using the fungus with pyridoxine marker, the TBR was operated continuously for 18 days and achieved a XynB output of 41 U/ml with an influent and effluent flow rate of 0.5 ml/min and a recycle flow rate of 56 ml/min. Production yields in the TBR were 1.4 times greater than a static tray culture and between 1.1 and 67 times greater than yields for SSF enzyme production stated in the literature. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Suitable conditions for xylanases activities from Bacillus sp. GA2(1 and Bacillus sp. GA1(6 and their properties for agricultural residues hydrolysis

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    Sudathip Chantorn

    2016-04-01

    Full Text Available Bacillus sp. GA2(1 and Bacillus sp. GA1(6 were isolated from soybean field in Khon Kaen province, Thailand. Crude enzymes from both isolates showed the activities of cellulase, xylanase, and mannanase at 37°C for 24 h. The highest xylanase activities of Bacillus sp. GA2(1 and Bacillus sp. GA1(6 were 1.58±0.25 and 0.82±0.16 U/ml, respectively. The relative xylanase activities from both strains were more than 60% at pH 5.0 to 8.0. The optimum temperature of xylanases was 50°C in both strains. The residual xylanase activities from both strains were more than 70% at 60°C for 60 min. Five agricultural wastes (AWs, namely coffee residue, soybean meal, potato peel, sugarcane bagasse, and corn cobs, were used as substrates for hydrolysis properties. The highest reducing sugar content of 101±1.32 µg/ml was obtained from soybean meal hydrolysate produced by Bacillus sp. GA2(1 xylanase.

  3. Effect of combined xylanase and phytase on growth performance, apparent total tract digestibility, and carcass characteristics in growing pigs fed corn-based diets containing high-fiber coproducts.

    Science.gov (United States)

    Jang, Y D; Wilcock, P; Boyd, R D; Lindemann, M D

    2017-09-01

    Phytate has been shown to be an antinutrient, and the feeding of high levels of phytase can break down phytate to improve nutrient utilization and pig performance. Dietary xylanase targets arabinoxylan breakdown, thereby improving energy utilization in pigs. However, the effects of simultaneous supplementation have not been clearly determined. Crossbred pigs ( = 45; mean initial weight, 26.4 ± 0.2 kg) were allotted to 1 of 9 treatments to evaluate the effects of both xylanase (endo-1,4-β xylanase [EC 3.2.1.8]) and phytase (6-phytase [EC 3.1.3.26]) supplementation as follows: 1) positive control (PC), a corn-soybean meal-based diet with 15% corn distillers dried grains with solubles, 15% wheat middlings, and 13% corn germ meal; 2) negative control (NC), ME was reduced by 103 kcal/kg from the PC diet by replacement of fat with corn starch; 3) NC + phytase (500 phytase units (FTU)/kg diet); 4) NC + phytase (1,000 FTU/kg diet); 5) NC + phytase (2,000 FTU/kg diet); 6) NC + xylanase (24,000 xylanase units [BXU]/kg diet); 7) NC + phytase (500 FTU/kg diet) + xylanase (24,000 BXU/kg diet); 8) NC + phytase (1,000 FTU/kg diet) + xylanase (24,000 BXU/kg diet); and 9) NC + phytase (2,000 FTU/kg diet) + xylanase (24,000 BXU/kg diet). All diets were formulated to meet nutrient requirements before phytase and xylanase addition to the diets. There were no significant interactions between xylanase and phytase supplementation on growth performance, carcass characteristics, and apparent total tract digestibility (ATTD). The ADG ( phytase level increased. The ATTD of P increased as phytase supplementation level increased ( phytase level increased. Estimated carcass lean percentage and lean gain increased ( phytase level increased. Xylanase supplementation had no effect on growth performance, ATTD, and carcass characteristics. The results demonstrated an improved nutrient digestibility, performance, and carcass response to phytase supplementation beyond P provision because all diets

  4. Concurrent production of cellulase and xylanase from Trichoderma reesei NCIM 1186: enhancement of production by desirability-based multi-objective method.

    Science.gov (United States)

    Jampala, Preethi; Tadikamalla, Satish; Preethi, M; Ramanujam, Swathy; Uppuluri, Kiran Babu

    2017-05-01

    Application of multiple response optimizations using desirability function in the production of microbial metabolites improves economy and efficiency. Concurrent production of cellulase and xylanase in Trichoderma reesei NCIM 1186 using an agricultural weed, Prosopis juliflora pods, was studied. The main aim of the study was to optimize significant medium nutrient parameters for maximization of cellulase and xylanase by multi-objective optimization strategy using biomass. Process parameters such as the nutrient concentrations (pods, sucrose, and yeast extract) and pH were investigated to improve cellulase and xylanase activities by one factor at a time approach, single response optimization and multi-objective optimization. At the corresponding optimized process parameters in single response optimization, the maximum cellulase activity observed was 3055.65 U/L where xylanase highest activity was 422.16 U/L. Similarly, the maximum xylanase activity, 444.94 U/L, was observed with the highest cellulase activity of 2804.40 U/L. The multi-objective optimization finds a tradeoff between the two objectives and optimal activity values in between the single-objective optima were achieved, 3033.74 and 439.13 U/L for cellulase and xylanase, respectively.

  5. Xylanase production by Trichoderma longibrachiatum

    Energy Technology Data Exchange (ETDEWEB)

    Royer, J C; Nakas, J P [State Univ. of New York, Syracuse, NY (USA). Coll. of Environmental Science and Forestry

    1989-07-01

    Xylan comprises up to 25% of hardwood biomass and is found in a variety of agricultural residues. It therefore represents a significant renewable resource which should be utilized to improve the economics of bioconversion of plant biomass to useful products. Before fermentation to fuels or solvents, xylan must be hydrolysed to xylose or short-chain oligomers of xylose. The effects of carbon source, substrate concentration, culture pH, and spore inoculum concentration on production of extracellular xylanase and cellulase were examined. Very low enzyme activities were obtained with growth on glucose, xylose, and cellobiose, while significantly higher levels were produced from lactose and arabinose. Higher levels of both enzymes were generated from alpha cellulose, wood pulp, and fibrous paper waste than from purified xylan. (author).

  6. Optimization of Xylanase Production through Response Surface Methodology by Fusarium sp. BVKT R2 Isolated from forest soil and its applications in saccharification

    Directory of Open Access Journals (Sweden)

    Ramanjaneyulu Golla

    2016-09-01

    Full Text Available AbstractXylanses are hydrolytic enzymes with wide applications in several industries like biofuels, paper and pulp, deinking, food and feed. The present study was aimed at hitting at high yield xylanase producing fungi from natural resources. Two highest xylanase producing fungal isolates - Q12 and L1were picked from collection of 450 fungal cultures for the utilization of xylan. These fungal isolates - Q12 and L1 were identified basing on ITS gene sequencing analysis as Fusarium sp. BVKT R2 (KT119615 and Fusarium strain BRR R6 (KT119619, respectively with construction of phylogenetic trees. Fusarium sp. BVKT R2 was further optimized for maximum xylanase production and the interaction effects between variables on production of xylanase were studied through response surface methodology. The optimal conditions for maximal production of xylanase were sorbitol 1.5%, yeast extract 1.5%, pH of 5.0, Temperature of 32.5ºC, and agitation of 175 rpm. Under optimal conditions, the yields of xylanase production by Fusarium sp. BVKT R2 was as high as 4560 U/ml in SmF. Incubation of different lignocellulosic biomasses with crude enzyme of Fusarium sp. BVKT R2 at 37°C for 72 h could achieve about 45% saccharification. The results suggest that Fusarium sp. BVKT R2 has potential applications in saccharification process of biomass.Key words: Fusarium sp., Optimization, Response Surface Methodology, Saccharification, Submerged fermentation, Xylanase

  7. CLONING, PURIFICATION AND CHARACTERIZATION OF HALOTOLERANT XYLANASE FROM Geobacillus Thermodenitrificans C5

    Directory of Open Access Journals (Sweden)

    Muhammad Irfan

    2016-06-01

    Full Text Available High levels of extracellular xylanase activity (994.50 IU/ml produced by Geobacillus thermodenitrificans C5 originated gene was detected when it was expressed in E. coli BL21 host. Thermostable xylanase (GthC5Xyl was purified to homogeneity and showed a molecular mass of approximately 44 kDa according to SDS-PAGE. The specific activity of the purified GthC5Xyl was up to 1243.125IU/mg with a 9.89-fold purification. The activity of GthC5Xyl was stimulated by CoCl2, MnSO4, CuSO4, MnCl2 but was inhibited by FeSO4, Hg, CaSO4. GthC5Xyl showed resistant to SDS, Tween 20, Triton X-100, β- Mercaptoethanol, PMSF, DTT, NEM and DEPC, SDS, and EDTA. A greater affinity for oat spelt xylan was exhibited by GthC5Xyl with maximum enzymatic activity at 60°C and 6.0 pH. The activity portrayed by GthC5Xyl was found to be acellulytic with stability at high temperature (70°C-80°C and low pH (4.0 to 8.0. Xylobiose and xylopentose were the end products of proficient oat spelt xylanase hydrolysis by GthC5Xyl. High SDS resistance and broader stability of GthC5Xyl proves it to be worthy of impending application in numerous industrial processes especially textile, detergents and animal feed industry.

  8. Structural Insights into the Thermophilic Adaption Mechanism of Endo-1,4-β-Xylanase from Caldicellulosiruptor owensensis.

    Science.gov (United States)

    Liu, Xin; Liu, Tengfei; Zhang, Yuebin; Xin, Fengjiao; Mi, Shuofu; Wen, Boting; Gu, Tianyi; Shi, Xinyuan; Wang, Fengzhong; Sun, Lichao

    2018-01-10

    Xylanases (EC 3.2.1.8) are a kind of enzymes degrading xylan to xylooligosaccharides (XOS) and have been widely used in a variety of industrial applications. Among them, xylanases from thermophilic microorganisms have distinct advantages in industries that require high temperature conditions. The CoXynA gene, encoding a glycoside hydrolase (GH) family 10 xylanase, was identified from thermophilic Caldicellulosiruptor owensensis and was overexpressed in Escherichia coli. Recombinant CoXynA showed optimal activity at 90 °C with a half-life of about 1 h at 80 °C and exhibited highest activity at pH 7.0. The activity of CoXynA activity was affected by a variety of cations. CoXynA showed distinct substrate specificities for beechwood xylan and birchwood xylan. The crystal structure of CoXynA was solved and a molecular dynamics simulation of CoXynA was performed. The relatively high thermostability of CoXynA was proposed to be due to the increased overall protein rigidity resulting from the reduced length and fluctuation of Loop 7.

  9. Enhanced sugar production from pretreated barley straw by additive xylanase and surfactants in enzymatic hydrolysis for acetone-butanol-ethanol fermentation.

    Science.gov (United States)

    Yang, Ming; Zhang, Junhua; Kuittinen, Suvi; Vepsäläinen, Jouko; Soininen, Pasi; Keinänen, Markku; Pappinen, Ari

    2015-01-01

    This study aims to improve enzymatic sugar production from dilute sulfuric acid-pretreated barley straw for acetone-butanol-ethanol (ABE) fermentation. The effects of additive xylanase and surfactants (polyethylene glycol [PEG] and Tween) in an enzymatic reaction system on straw hydrolysis yields were investigated. By combined application of 2g/100g dry-matter (DM) xylanase and PEG 4000, the glucose yield was increased from 53.2% to 86.9% and the xylose yield was increased from 36.2% to 70.2%, which were considerably higher than results obtained with xylanase or surfactant alone. The ABE fermentation of enzymatic hydrolysate produced 10.8 g/L ABE, in which 7.9 g/L was butanol. The enhanced sugar production increased the ABE yield from 93.8 to 135.0 g/kg pretreated straw. The combined application of xylanase and surfactants has a large potential to improve sugar production from barley straw pretreated with a mild acid and that the hydrolysate showed good fermentability in ABE production. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Effect of Aspergillus niger xylanase on dough characteristics and bread quality attributes.

    Science.gov (United States)

    Ahmad, Zulfiqar; Butt, Masood Sadiq; Ahmed, Anwaar; Riaz, Muhammad; Sabir, Syed Mubashar; Farooq, Umar; Rehman, Fazal Ur

    2014-10-01

    The present study was conducted to investigate the impact of various treatments of xylanase produced by Aspergillus niger applied in bread making processes like during tempering of wheat kernels and dough mixing on the dough quality characteristics i.e. dryness, stiffness, elasticity, extensibility, coherency and bread quality parameters i.e. volume, specific volume, density, moisture retention and sensory attributes. Different doses (200, 400, 600, 800 and 1,000 IU) of purified enzyme were applied to 1 kg of wheat grains during tempering and 1 kg of flour (straight grade flour) during mixing of dough in parallel. The samples of wheat kernels were agitated at different intervals for uniformity in tempering. After milling and dough making of both types of flour (having enzyme treatment during tempering and flour mixing) showed improved dough characteristics but the improvement was more prominent in the samples receiving enzyme treatment during tempering. Moreover, xylanase decreased dryness and stiffness of the dough whereas, resulted in increased elasticity, extensibility and coherency and increase in volume & decrease in bread density. Xylanase treatments also resulted in higher moisture retention and improvement of sensory attributes of bread. From the results, it is concluded that dough characteristics and bread quality improved significantly in response to enzyme treatments during tempering as compared to application during mixing.

  11. Optimization of Cellulase and Xylanase Production by Micrococcus Species under Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Ziyanda Mmango-Kaseke

    2016-11-01

    Full Text Available This paper reports on the optimization of culture conditions for cellulase and xylanase production by bacterial isolate from lignocellulosic biomass. The bacterial isolate was screened for cellulase and xylanase production on carboxyl methyl cellulose (CMC and birch wood xylan as substrates, respectively. One bacterial isolate showing the highest halo zone diameter (isolate PLY1 was selected for detailed studies. The analysis of the 16S ribosomal ribonucleic acid (rRNA gene nucleotide sequence of PLY1 revealed it to have 98% similarity to Micrococcus luteus strain Fse9 and the sequence was deposited in the GenBank as Micrococcus luteus strain SAMRC-UFH3 with accession number KU171371. Cellulase production was achieved in the presence of CMC (1% w/v under an incubation temperature of 25 °C (198 U/mL, pH 5 (173 U/mL, agitation speed 50 rpm (173 U/mL and incubation period of 96 h (102 U/mL. Xylanase was produced maximally when birch wood xylan (1% w/v was used as the substrate at 25 °C (1007 U/mL, pH 10 (2487 U/mL, 200 rpm (1814 U/mL, and under an incubation period of 84 h (1296 U/mL. Our findings showed that Micrococcus sp. SAMRC-UFH3 appears to be a potentially important candidate for lignocellulosic waste degradation and other relevant industrial applications.

  12. Influence of xylanase addition on the characteristics of loaf bread prepared with white flour or whole grain wheat flour

    Directory of Open Access Journals (Sweden)

    Leandra Zafalon Jaekel

    2012-12-01

    Full Text Available The aim of this study was to verify the influence of the addition of the enzyme xylanase (four concentrations: 0, 4, 8, and 12 g.100 kg-1 flour on the characteristics of loaf bread made with white wheat flour or whole grain wheat flour. Breads made from white flour and added with xylanase had higher specific volumes than those of the control sample (no enzyme; however, the specific volume did not differ significantly (p < 0.05 among the breads with different enzyme concentrations. All formulations made from whole grain wheat flour and added with xylanase also had specific volumes significantly higher than those of the control sample, and the highest value was found for the 8 g xylanase.100 kg-1 flour formulation. With respect to moisture content, the formulations with different enzyme concentrations showed small significant differences when compared to the control samples. In general, breads made with the addition of 8 g enzyme.100 kg-1 flour had the lowest firmness values, thus presenting the best technological characteristics.

  13. Cloning, expression, purification, and characterisation of the HEAT-repeat domain of TOR from the thermophilic eukaryote Chaetomium thermophilum.

    Science.gov (United States)

    Robinson, Graham C; Vegunta, Yogesh; Gabus, Caroline; Gaubitz, Christl; Thore, Stéphane

    2017-05-01

    The Target of Rapamycin Complex is a central controller of cell growth and differentiation in eukaryotes. Its global architecture has been described by cryoelectron microscopy, and regions of its central TOR protein have been described by X-ray crystallography. However, the N-terminal region of this protein, which consists of a series of HEAT repeats, remains uncharacterised at high resolution, most likely due to the absence of a suitable purification procedure. Here, we present a robust method for the preparation of the HEAT-repeat domain, utilizing the thermophilic fungus Chaetomium thermophilum as a source organism. We describe construct design and stable expression in insect cells. An efficient two-step purification procedure is presented, and the purified product is characterised by SEC and MALDI-TOF MS. The methods described pave the way for a complete high-resolution characterisation of this elusive region of the TOR protein. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Aspects microbiologiques de la production par fermentation solide des endo-beta-1,4-xylanases de moisissures : le cas de Penicillium canescens

    Directory of Open Access Journals (Sweden)

    Assamoi AA.

    2009-01-01

    Full Text Available Microbial aspects of endo-β-1,4-xylanase production in solid-state fermentation by Penicillia: the case of Penicillium canescens. Production of xylanases by Penicillium canescens 10-10c is the research object in Walloon Center of Industrial Biology. Previous works used submerged or liquid fermentation. The actual works are oriented more and more towards solid fermentation from agricultural or agro-alimentary residues. In addition to the valorization of these residues, solid-state fermentation reaches an increasingly significant interest in various other fields like the biological breakdown of the solid residues, the bioremediation of the organic pollutants in the grounds and the reduction of the air pollution by the biofiltration. Xylanase is an industrial enzyme used in general in extraction and clarification processes. P. canescens can produce an activity of it, particularly in its balanced forms of xylanases, beta-xylosidase and arabinosidase, and not contaminated by cellulolytic and amylolytic activities. It is a hyper producing strain of xylanase. The production rate is one of the highest in literature (535 U.ml-1 and 9,632 U.g-1 in Erlenmeyer flasks, in submerged and solid state fermentation, respectively. The biobleaching activity of the cellulose pulp by the purified enzyme is higher than a commercial preparation of xylanases from Trichoderma longibrachiatum used industrially. It has a complete hydrolysis degree of 40% (on glucuronoxylan and 35% (on arabinoxylan at 55°C and at pH of 5.9. These characteristics lead to many industrial applications of this enzyme. That is why the optimization of its production by the solid-state fermentation at the laboratory scale in order to define a policy for the industrial transposition later is carried out. This article presents a summary of the scientific literature on this subject.

  15. The Influence of Xylanase Supplementation on Dough Rheology Concerning its Consistograph Parameters

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    Rodica Chereji

    2010-05-01

    Full Text Available In this study we determined the influence of xylanase supplementation on dough rheology concerning its consistograph parameters: maximum pressure (Pr max, (mb and water absorption (Wa, %. The consistograph analyses were conducted at constant hydration and consistency of 500UF. Determinations were made on 4 types of flour and optimal enzyme dosages were determined. Then we added the optimal enzyme dose for each type of flour as follows: F1, F2, F3, F4: P1-8100U.FXU/100kg flour, P2-16200U.FXU/100kg flour, P3-24300U.FXU/100kg flour. Fungal xylanase used in these concentrations led to the improvement of bread quality properties: finer texture of the crumb, extending freshness of bread, improving the colour and flavour, improving the slicing ability.

  16. Xylanase production from Thermomyces lanuginosus VAPS-24 using low cost agro-industrial residues via hybrid optimization tools and its potential use for saccharification.

    Science.gov (United States)

    Kumar, Vishal; Chhabra, Deepak; Shukla, Pratyoosh

    2017-11-01

    The xylanase production from Thermomyces lanuginosus VAPS-24 has been optimized using OFAT (One factor at a time) approach using agro-industrial substrates. Further, central composite design (CCD) has been employed to optimize various process parameters such as temperature (45-55°C), carbon source concentration (1.5-2.5%), fermentation time (72-120h) and production medium pH (6-8). Maximum xylanase yield after RSM optimization was approximately double (119.91±2.53UmL -1 ) than un-optimized conditions (61.09±0.91UmL -1 ). Several hybrid statistical tools such as Genetic Algorithm-Response Surface Methodology (GA-RSM), Artificial Neural Network (ANN), Genetic Algorithm-Artificial Neural Network (GA-ANN) were employed to obtain more optimized process parameters to maximize the xylanase production and observed an increase of 10.50% xylanase production (132.51±3.27UmL -1 ) as compared to RSM response (119.91±2.53UmL -1 ). The various pretreated and untreated agricultural residues were subjected to saccharification by using crude xylanase in which the pretreated rice straw yielded maximum fermentable sugars 126.89mgg -1 . Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. The influence of sorbitol on the production of cellulases and xylanases in an airlift bioreactor.

    Science.gov (United States)

    Ritter, Carla Eliana Todero; Fontana, Roselei Claudete; Camassola, Marli; da Silveira, Maurício Moura; Dillon, Aldo José Pinheiro

    2013-11-01

    The production of cellulases and xylanases by Penicillium echinulatum in an airlift bioreactor was evaluated. In batch production, we tested media with isolated or associated cellulose and sorbitol. In fed-batch production, we tested cellulose addition at two different times, 30 h and 48 h. Higher liquid circulation velocities in the downcomer were observed in sorbitol 10 g L(-1) medium. In batch production, higher FPA (filter paper activity) and endoglucanase activities were obtained with cellulose (7.5 g L(-1)) and sorbitol (2.5 g L(-1)), 1.0 U mL(-1) (120 h) and 6.4 U m L(-1) (100 h), respectively. For xylanases, the best production condition was cellulose 10 g L(-1), which achieved 5.5 U mL(-1) in 64 h. The fed-batch process was favorable for obtaining xylanases, but not for FPA and endoglucanases, suggesting that in the case of cellulases, the inducer must be added early in the process. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. Heterologous expression of xylanase enzymes in lipogenic yeast Yarrowia lipolytica.

    Directory of Open Access Journals (Sweden)

    Wei Wang

    Full Text Available To develop a direct microbial sugar conversion platform for the production of lipids, drop-in fuels and chemicals from cellulosic biomass substrate, we chose Yarrowia lipolytica as a viable demonstration strain. Y. lipolytica is known to accumulate lipids intracellularly and is capable of metabolizing sugars to produce lipids; however, it lacks the lignocellulose-degrading enzymes needed to break down biomass directly. While research is continuing on the development of a Y. lipolytica strain able to degrade cellulose, in this study, we present successful expression of several xylanases in Y. lipolytica. The XynII and XlnD expressing Yarrowia strains exhibited an ability to grow on xylan mineral plates. This was shown by Congo Red staining of halo zones on xylan mineral plates. Enzymatic activity tests further demonstrated active expression of XynII and XlnD in Y. lipolytica. Furthermore, synergistic action in converting xylan to xylose was observed when XlnD acted in concert with XynII. The successful expression of these xylanases in Yarrowia further advances us toward our goal to develop a direct microbial conversion process using this organism.

  19. The enhancement of enzymatic hydrolysis of lignocellulosic substrates by the addition of accessory enzymes such as xylanase: is it an additive or synergistic effect?

    Directory of Open Access Journals (Sweden)

    Saddler Jack N

    2011-10-01

    Full Text Available Abstract Background We and other workers have shown that accessory enzymes, such as β-glucosidase, xylanase, and cellulase cofactors, such as GH61, can considerably enhance the hydrolysis effectiveness of cellulase cocktails when added to pretreated lignocellulosic substrates. It is generally acknowledged that, among the several factors that hamper our current ability to attain efficient lignocellulosic biomass conversion yields at low enzyme loadings, a major problem lies in our incomplete understanding of the cooperative action of the different enzymes acting on pretreated lignocellulosic substrates. Results The reported work assessed the interaction between cellulase and xylanase enzymes and their potential to improve the hydrolysis efficiency of various pretreated lignocellulosic substrates when added at low protein loadings. When xylanases were added to the minimum amount of cellulase enzymes required to achieve 70% cellulose hydrolysis of steam pretreated corn stover (SPCS, or used to partially replace the equivalent cellulase dose, both approaches resulted in enhanced enzymatic hydrolysis. However, the xylanase supplementation approach increased the total protein loading required to achieve significant improvements in hydrolysis (an additive effect, whereas the partial replacement of cellulases with xylanase resulted in similar improvements in hydrolysis without increasing enzyme loading (a synergistic effect. The enhancement resulting from xylanase-aided synergism was higher when enzymes were added simultaneously at the beginning of hydrolysis. This co-hydrolysis of the xylan also influenced the gross fiber characteristics (for example, fiber swelling resulting in increased accessibility of the cellulose to the cellulase enzymes. These apparent increases in accessibility enhanced the steam pretreated corn stover digestibility, resulting in three times faster cellulose and xylan hydrolysis, a seven-fold decrease in cellulase loading and

  20. Preservation of Bacillus pumilus PU4-2 xylanases by immobilization technique into pollard and cation addition

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    T Haryati

    2010-03-01

    Full Text Available Utilization of by-product from agriculture as alternative source of feedstuff has been widely practiced. However their usage is limited due to high fiber content and low nutrient digestibility. The use of specific hydrolizing enzymes, xylanases are gaining importance because of their wide application in various industrial sectors especially in bioconversion of hemicellulosic material. This experiment was done to evaluate the effect of cation addition and immobilization of enzyme into pollard on stability of B. pumilus xylanase. The enzyme extract was purified by precipitation with 75% ammonium sulphate. Four kinds of cation (Ca2+, Fe3+, Mg2+, Zn2+ were added to the purified enzyme, at concentration of 1m M and stored at 4 and 27˚C. For immobilization process, the optimum enzyme concentration that will be added to pollard has been evaluated by analysis of xylanase activity and their recovery. The specific activity of enzyme after precipitation increased 1.8 times, from 420.3 to 765.2 U/mg protein. All cations act as activator which relative activity become 130.6; 139.0; 103.8 and 163.5% respectively. Concentration of 0.5mM Ca2+ and Fe3+ were most able to keep xylanases activity stable at 4˚C. The optimum composition of enzymes and pollard was 1.5 ml for 5 gram of pollard with recovery of xylanases activity of 82.2%. In immobilized enzyme, the activity of enzyme without cation addition is higher than that with addition of Ca2+ and Fe3+. Activity of enzyme stored at 4˚C is more stable than that at 27˚C. Immobilized enzyme is more stable for storage, which lasted for 7 weeks at 27˚C and 12 weeks at 4˚C compared to liquid enzyme which lasted for only 7 days at 27˚C and 13 days at 4˚C.

  1. Wheat dough rheology at low water contents and the influence of xylanases

    NARCIS (Netherlands)

    Hardt, N.A.; Boom, R.M.; Goot, van der A.J.

    2014-01-01

    The effect of low water contents and xylanases on wheat dough rheology is reported. Farinograph, dynamic oscillation, and creep-recovery measurements were performed using water concentrations from 34 to 44.8% (total basis). A water reduction from 43.5–44.8% to 34% increased resistance upon mixing as

  2. Characterization and mode of action of xylanases ␁and some accessory enzymes

    NARCIS (Netherlands)

    Kormelink, F.J.M.

    1992-01-01

    Three endo-(l,4)-β-D-xylanases; (Endo I, Endo II, and Endo III), a (1,4)-β-xylosidase and an (1,4)-β-D-arabinoxylan arabinofuranohydrolase (AXH) were purified from a culture filtrate produced by Aspergillus awamori CMI 142717. In addition to these enzymes, an acetyl

  3. Production of spent mushroom substrate hydrolysates useful for cultivation of Lactococcus lactis by dilute sulfuric acid, cellulase and xylanase treatment.

    Science.gov (United States)

    Qiao, Jian-Jun; Zhang, Yan-Fei; Sun, Li-Fan; Liu, Wei-Wei; Zhu, Hong-Ji; Zhang, Zhijun

    2011-09-01

    Spent mushroom substrate (SMS) was treated with dilute sulfuric acid followed by cellulase and xylanase treatment to produce hydrolysates that could be used as the basis for media for the production of value added products. A L9 (3(4)) orthogonal experiment was performed to optimize the acid treatment process. Pretreatment with 6% (w/w) dilute sulfuric acid at 120°C for 120 min provided the highest reducing sugar yield of 267.57 g/kg SMS. No furfural was detected in the hydrolysates. Exposure to 20PFU of cellulase and 200 XU of xylanase per gram of pretreated SMS at 40°C resulted in the release of 79.85 g/kg or reducing sugars per kg acid pretreated SMS. The dilute sulfuric acid could be recycled to process fresh SMS four times. SMS hydrolysates neutralized with ammonium hydroxide, sodium hydroxide, or calcium hydroxide could be used as the carbon source for cultivation of Lactococcus lactis subsp. lactis W28 and a cell density of 2.9×10(11)CFU/mL could be obtained. The results provide a foundation for the development of value-added products based on SMS. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. ETHANOL PRECIPITATION OF GLYCOSYL HYDROLASES PRODUCED BY Trichoderma harzianum P49P11

    Directory of Open Access Journals (Sweden)

    M. A. Mariño

    2015-06-01

    Full Text Available AbstractThis study aimed to concentrate glycosyl hydrolases produced by Trichoderma harzianum P49P11 by ethanol precipitation. The variables tested besides ethanol concentration were temperature and pH. The precipitation with 90% (v/v ethanol at pH 5.0 recovered more than 98% of the xylanase activity, regard less of the temperature (5.0, 15.0, or 25.0 °C. The maximum recovery of cellulase activity as FPase was 77% by precipitation carried out at this same pH and ethanol concentration but at 5.0 °C. Therefore, ethanol precipitation can be considered to be an efficient technique for xylanase concentration and, to a certain extent, also for the cellulase complex.

  5. Production of cellulase and xylanase in a bubble gum column using immobilized Aspergillus niger KKS

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Seong-Woo; Kim, Seung-Woo [Univ. of Suwon (Korea, Republic of); Lee, Jin-Suk [Korea Institute of Energy Research, Daejeon (Korea, Republic of)

    1995-05-01

    Aspergillus niger KKS, isolated from a farmland near Suwon, was immobilized on Celite and polyurethane foams. Enzyme activities produced by the immobilized cell system in a bubble column were higher than that of shake-flask culture. The enzyme productivities were twice as high. {Beta}-Glucosidase, {Beta}-xylosidase, and xylanase activities obtained in a bubble column were significant when the ground rice straw was used as a substrate. 9 refs., 2 figs., 3 tabs.

  6. Cellulase and xylanase productions by isolated Amazon Bacillus strains using soybean industrial residue based solid-state cultivation

    Directory of Open Access Journals (Sweden)

    Heck Júlio X.

    2002-01-01

    Full Text Available In Brazil, a large amount of a fibrous residue is generated as result of soybean (Glycine max protein production. This material, which is rich in hemicellulose and cellulose, can be used in solid state cultivations for the production of valuable metabolites and enzymes. In this work, we studied the bioconversion of this residue by bacteria strains isolated from water and soil collected in the Amazon region. Five strains among 87 isolated bacteria selected for their ability to produce either celullases or xylanases were cultivated on the aforementioned residue. From strain BL62, identified as Bacillus subtilis, it was obtained a preparation showing the highest specific cellulase activity, 1.08 UI/mg protein within 24 hours of growth. Concerning xylanase, the isolate BL53, also identified as Bacillus subtilis, showed the highest specific activity for this enzyme, 5.19 UI/mg protein within 72 hours of cultivation. It has also been observed the production of proteases that were associated with the loss of cellulase and xylanase activities. These results indicated that the selected microorganisms, and the cultivation process, have great biotechnological potential.

  7. Co-cultivation of mutant Penicillium oxalicum SAU(E)-3.510 and Pleurotus ostreatus for simultaneous biosynthesis of xylanase and laccase under solid-state fermentation.

    Science.gov (United States)

    Dwivedi, Pallavi; Vivekanand, V; Pareek, Nidhi; Sharma, Amit; Singh, Rajesh P

    2011-10-01

    Co-cultivation of mutant Penicillium oxalicum SAU(E)-3.510 and Pleurotus ostreatus MTCC 1804 was evaluated for the production of xylanase-laccase mixture under solid-state fermentation (SSF) condition. Growth compatibility between mutant P. oxalicum SAU(E)-3.510 and white rot fungi (P. ostreatus MTCC 1804, Trametes hirsuta MTCC 136 and Pycnoporus sp. MTCC 137) was analyzed by growing them on potato dextrose agar plate. Extracellular enzyme activities were determined spectrophotometrically. Under derived conditions, paired culturing of mutant P. oxalicum SAU(E)-3.510 and P. ostreatus MTCC 1804 resulted in 58% and 33% higher levels of xylanase and laccase production, respectively. A combination of sugarcane bagasse and black gram husk in a ratio of 3:1 was found to be the most ideal solid substrate and support for fungal colonization and enzyme production during co-cultivation. Maximum levels of xylanase (8205.31 ± 168.31 IU g(-1)) and laccase (375.53 ± 34.17 IU g(-1)) during SSF were obtained by using 4 g of solid support with 80% of moisture content. Furthermore, expressions of both xylanase and laccase were characterized during mixed culture by zymogram analysis. Improved levels of xylanase and laccase biosynthesis were achieved by co-culturing the mutant P. oxalicum SAU(E)-3.510 and P. ostreatus MTCC 1804. This may be because of efficient substrate utilization as compared to their respective monocultures in the presence of lignin degradation compounds because of synergistic action of xylanase and laccase. Understanding and developing the process of co-cultivation appears productive for the development of mixed enzyme preparation with tremendous potential for biobleaching. Copyright © 2011 Elsevier B.V. All rights reserved.

  8. Application of xylanases from Amazon Forest fungal species in bleaching of eucalyptus kraft pulps

    Directory of Open Access Journals (Sweden)

    Roseli Garcia Medeiros

    2007-03-01

    Full Text Available Crude xylanase preparations from Penicillium corylophilum, Aspergillus niger and Trichoderma longibrachiatum were used to treat Eucalyptus kraft pulp, prior to chlorine dioxide and alkaline bleaching sequences. The enzyme pretreatment improved brightness and delignification of non-delignified and oxygen-bleached samples of eucalyptus kraft pulp. Xylanase preparations from T. longibrachiatum and P. corylophilum were more effective to reduce pulp kappa number. A small reduction in viscosity was obtained when the oxygen-bleached pulp was treated with xylanase preparation from A. niger. For all enzyme samples, the best release of chromophoric material from the pulp was at 237 nm. The enzyme preparation from P. corylophilum was responsible for the highest release of reducing sugar at a dosage interval of 10-20 IU/g dry weight pulp. Scanning electron microscopy studies of oxygen-bleached pulp after xylanase treatment revealed morphological changes, including holes, cracks, filament forming and peeling.Amostras de xilanases de extratos brutos de Penicillium corylophilum, Aspergillus niger e Trichoderma longibrachiatum foram utilizadas no branqueamento de polpa kraft de eucalipto antes das seqüências alcalina e dióxido de cloro. O pré-tratamento enzimático melhorou a alvura e o processo de deslignificação de amostras de polpa kraft de eucalipto não-tratada e tratada com oxigênio. Amostras de xilanases de T. longibrachiatum e P. corylophilum foram mais efetivas na redução do número kappa da polpa. A polpa tratada com oxigênio sofreu uma pequena redução na sua viscosidade quando incubada com amostra de xilanase de A. niger. Para todas as amostras de xilanases, a maior liberação de cromóforos da polpa foi a 237 nm. A amostra de xilanase de P. corylophilum liberou maior quantidade de açúcar redutor da polpa, utilizando dosagem de 10-20 UI/g de peso seco da polpa. Estudos de microscopia eletrônica de varredura revelaram várias altera

  9. Improvement of bread making quality by supplementation with a recombinant xylanase produced by Pichia pastoris.

    Science.gov (United States)

    de Queiroz Brito Cunha, Carolina Cândida; Gama, Aline Rodrigues; Cintra, Lorena Cardoso; Bataus, Luiz Artur Mendes; Ulhoa, Cirano José

    2018-01-01

    Xylanases (EC 3.2.1.8) are hydrolytic enzymes, which randomly cleave the β-1,4-linked xylose residues from xylan. The synthetic gene xynBS27 from Streptomyces sp. S27 was successfully cloned and expressed in Pichia pastoris. The full-length gene consists of 729 bp and encodes 243 amino acids including 51 residues of a putative signal peptide. This enzyme was purified in two steps and was shown to have a molecular weight of 20 kDa. The purified r-XynBS27 was active against beechwood xylan and oat spelt xylan as expected for GH 11 family. The optimum pH and temperature values for the enzyme were 6.0 and 75 °C, respectively. The Km and Vmax were 12.38 mg/mL and 13.68 μmol min/mg, respectively. The r-XynBS27 showed high xylose tolerance and was inhibited by some metal ions and by SDS. r-XynBS27 was employed as an additive in the bread making process. A decrease in firmness, stiffness and consistency, and improvements in specific volume and reducing sugar content were recorded.

  10. Purification and characterization of five cellulases and one xylanase from Penicillium brasilianum IBT 20888

    DEFF Research Database (Denmark)

    Jørgensen, Henning; Eriksson, T.; Borjesson, J.

    2003-01-01

    The filamentous fungus Penicillium brasilianum IBT 20888 was cultivated on a mixture of 30 g l(-1) cellulose and 10 g l(-1) xylan for 111 h and the resulting culture filtrate was used for protein purification. From the cultivation broth, five cellulases and one xylanase were purified. Hydrolysis...... studies revealed that two of the cellulases were acting as cellobiohydrolases by being active on only microcrystalline cellulose (Avicel). Three of the cellulases were active on both Avicel and carboxymethyl cellulose indicating endoglucanase activity. Two of these showed furthermore mannanase activity...... the cellulose-binding domain or an essential part of it. The basic xylanase (pI > 9) was only active towards xylan. Two of the purified cellulases with endoglucanase activity were partly sequenced and based on sequence homology with known enzymes they were classified as belonging to families 5 and 12...

  11. Concomitant production of cellulase and xylanase by thermophilic mould Sporotrichum thermophile in solid state fermentation and their applicability in bread making.

    Science.gov (United States)

    Bala, Anju; Singh, Bijender

    2017-06-01

    Sporotrichum thermophile BJAMDU5 secreted high titres of xylanolytic and cellulolytic enzymes in solid state fermentation using mixture of wheat straw and cotton oil cake (ratio 1:1) at 45 °C, pH 5.0 after 72 h inoculated with 2.9 × 10 7  CFU/mL conidiospores. Supplementation of solid medium with lactose and ammonium sulphate further enhanced the production of hydrolytic enzymes. Among different surfactants studied, Tween 80 enhanced the production of all enzymes [3455 U/g DMR (dry mouldy residue), 879.26 U/g DMR, 976.28 U/g DMR and 35.10 U/g DMR for xylanase, CMCase (Carboxymethylcellulase), FPase (Filter paper activity) and β-glucosidase, respectively] as compared to other surfactants. Recycling of solid substrate reduced the production of all these enzymes after second cycle. End products analysis by TLC showed the ability of hydrolytic enzymes of S. thermophile to liberate monomeric (xylose and glucose) as well as oligomeric (xylobiose, cellobiose and higher ones) sugars. Supplementation of enzyme resulted in improved nutritional properties of the bread. Formation of oligomeric sugars by xylanase enzyme of S. thermophile BJAMDU5 make it a good candidate in food industry.

  12. Parthenium sp. as a plant biomass for the production of alkalitolerant xylanase from mutant Penicillium oxalicum SAUE-3.510 in submerged fermentation

    International Nuclear Information System (INIS)

    Dwivedi, Pallavi; Vivekanand, V.; Ganguly, Ruma; Singh, Rajesh P.

    2009-01-01

    The use of congress grass (Parthenium sp.) and water hyacinth (Eichhornia crassipes) as low cost raw materials for xylanase production from mutant Penicillium oxalicum SAU E -3.510 in submerged fermentation was investigated. For development of mutant from wild type P. oxalicum SA-8 ITCC 6024, a strategy of mixed mutagenesis was followed using UV-irradiation and ethidium bromide, which had resulted into 1.87 fold increases in the activity of the enzyme. For enzyme production, the fungus was cultivated in mineral medium containing congress grass as carbon source. Considerably higher levels of production (475.2 ± 6.0 IU ml -1 ) were achieved in media containing congress grass, although it was slightly less than that was obtained (488.5 ± 6.5 IU ml -1 ) in presence of commercial oat spelt xylan. This fact confirms the feasibility of using this low cost non-food resource as an alternative carbon source to save costs of the enzyme production process. Maximum xylanase activity was reported at 55 deg. C with its stability at 80 deg. C for 2 h. The highest activity of xylanase at pH 9.0 and its stability at similar pH for 24 h denote the alkalitolerant nature of enzyme

  13. An explanation for the combined effect of xylanase-glucose oxidase in dough systems

    NARCIS (Netherlands)

    Primo-Martín, C.; Wang, M.; Lichtendonk, W.J.; Plijter, J.J.; Hamer, R.J.

    2005-01-01

    In the bakery industry, glucose oxidase is usually used in combination with xylanase. Although many theories exist on the mechanism of action of each enzyme, the positive effect of combining the two is as yet unexplained. In this paper we studied a possible basis for this synergy by focusing on the

  14. Rapid 2,2'-bicinchoninic-based xylanase assay compatible with high throughput screening

    Science.gov (United States)

    William R. Kenealy; Thomas W. Jeffries

    2003-01-01

    High-throughput screening requires simple assays that give reliable quantitative results. A microplate assay was developed for reducing sugar analysis that uses a 2,2'-bicinchoninic-based protein reagent. Endo-1,4-â-D-xylanase activity against oat spelt xylan was detected at activities of 0.002 to 0.011 IU ml−1. The assay is linear for sugar...

  15. Xylanase and Protease Increase Solubilization of Non-Starch Polysaccharides and Nutrient Release of Corn- and Wheat Distillers Dried Grains with Solubles

    DEFF Research Database (Denmark)

    Pedersen, Mads Brøgger; Dalsgaard, Søren; Arent, Susan

    2015-01-01

    The use of distiller dried grains with solubles (DDGS) as alternative to conventional animal feed for non-ruminants is challenged by the high content of non-starch polysaccharides and varying protein quality. In this study the enzymatic degradation of corn- and wheat DDGS was evaluated, in vitro...... of this xylanase. The current in vitro results indicate a high potential of xylanase in combination with protease to efficiently degrade DDGS and promote nutrient release in diets for non-ruminant animals....

  16. Bi-functional fusion enzyme EG-M-Xyn displaying endoglucanase and xylanase activities and its utility in improving lignocellulose degradation.

    Science.gov (United States)

    Chen, Chin-Chung; Gao, Guo-Jhan; Kao, Ai-Ling; Tsai, Zheng-Chia

    2018-05-01

    In this study, the gene fusion of endoglucanase (EG, one of cellulases) from Teleogryllus emma and xylanase (Xyn, one of hemicellulases) from Thermomyces lanuginosus was constructed to generate a fusion enzyme (EG-M-Xyn). Through the expression and purification by ultrafiltration and size-exclusion chromatography, the purified EG-M-Xyn had a molecular weight of 75.5 kDa and exhibited the specific activity of CMCase and xylanase as 306.8 U/mg and 1227.3 U/mg, respectively. The K m values (CMC and beechwood xylan) were 6.8 and 60.6 mg mL -1 while catalytic efficiency (k cat /K m ) values of CMCase and xylanase were 3280 and 38,797 min -1  mg -1  mL, respectively. EG-M-Xyn exerted great properties for its great potential in improving the enzymatic hydrolysis of lignocellulosics to produce fermentable sugars. First, EG-M-Xyn showed mild reaction pH and temperature of 5.5 and 50 °C, respectively. Secondly, EG-M-Xyn exhibited great heat tolerance of T 1/2 values of 173 (CMCase) and 693 min (xylanase). Lastly and most importantly, application of EG-M-Xyn in combination with Ctec2 (commercial enzyme) in the saccharification led to a 10-20% net increase in fermentable sugars liberated from pretreated rice straw in comparison to the Ctec2 alone group. In conclusion, EG-M-Xyn had great potential in generating fermentable sugars from renewable agro-residues for biofuel and fine chemical industry. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Thermophilic fungi as new sources for production of cellulases and xylanases with potential use in sugarcane bagasse saccharification.

    Science.gov (United States)

    de Cassia Pereira, J; Paganini Marques, N; Rodrigues, A; Brito de Oliveira, T; Boscolo, M; da Silva, R; Gomes, E; Bocchini Martins, D A

    2015-04-01

    To obtain new cellulases and xylanases from thermophilic fungi; evaluate their potential for sugarcane bagasse saccharification. Thirty-two heat-tolerant fungi were isolated from the environment, identified (morphological/molecular tools) and the production of the enzymes was evaluated by solid state fermentation using lignocellulosic materials as substrates. Myceliophthora thermophila JCP 1-4 was the best producer of endoglucanase (357·51 U g(-1) ), β-glucosidase (45·42 U g(-1) ), xylanase (931·11 U g(-1) ) and avicelase (3·58 U g(-1) ). These enzymes were most active at 55-70°C and stable at 30-60°C. Using crude enzymatic extract from M. thermophila JCP 1-4 to saccharify sugarcane bagasse pretreated with microwaves and glycerol, glucose and xylose yields obtained were 15·6 and 35·13% (2·2 and 1·95 g l(-1) ), respectively. All isolated fungi have potential to produce the enzymes; M. thermophila JCP 1-4 enzymatic extract have potential to be better explored in saccharification experiments. Pretreatment improved enzymatic saccharification, as sugar yields were much higher than those obtained from in natura bagasse. Myceliophthora thermophila JCP 1-4 produces avicelase (not commonly found among fungi; important to hydrolyse crystalline cellulose) and a β-glucosidase resistant to glucose inhibition, interesting characteristics for saccharification experiments. © 2015 The Society for Applied Microbiology.

  18. Robust expression and secretion of Xylanase1 in Chlamydomonas reinhardtii by fusion to a selection gene and processing with the FMDV 2A peptide.

    Directory of Open Access Journals (Sweden)

    Beth A Rasala

    Full Text Available Microalgae have recently received attention as a potential low-cost host for the production of recombinant proteins and novel metabolites. However, a major obstacle to the development of algae as an industrial platform has been the poor expression of heterologous genes from the nuclear genome. Here we describe a nuclear expression strategy using the foot-and-mouth-disease-virus 2A self-cleavage peptide to transcriptionally fuse heterologous gene expression to antibiotic resistance in Chlamydomonas reinhardtii. We demonstrate that strains transformed with ble-2A-GFP are zeocin-resistant and accumulate high levels of GFP that is properly 'cleaved' at the FMDV 2A peptide resulting in monomeric, cytosolic GFP that is easily detectable by in-gel fluorescence analysis or fluorescent microscopy. Furthermore, we used our ble2A nuclear expression vector to engineer the heterologous expression of the industrial enzyme, xylanase. We demonstrate that linking xyn1 expression to ble2A expression on the same open reading frame led to a dramatic (~100-fold increase in xylanase activity in cells lysates compared to the unlinked construct. Finally, by inserting an endogenous secretion signal between the ble2A and xyn1 coding regions, we were able to target monomeric xylanase for secretion. The novel microalgae nuclear expression strategy described here enables the selection of transgenic lines that are efficiently expressing the heterologous gene-of-interest and should prove valuable for basic research as well as algal biotechnology.

  19. Isolation, screening and characterization of a novel extracellular xylanase from Aspergillus niger (KP874102.1) and its application in orange peel hydrolysis.

    Science.gov (United States)

    Uday, Uma Shankar Prasad; Majumdar, Ria; Tiwari, Onkar Nath; Mishra, Umesh; Mondal, Abhijit; Bandyopadhyay, Tarun Kanti; Bhunia, Biswanath

    2017-12-01

    In the present work, a potent xylanase producing fungal strain Aspergillus niger (KP874102.1) was isolated through cultural and morphological observations from soil sample of Baramura forest, Tripura west, India. 28S rDNA technique was applied for genomic identification of this fungal strain. The isolated strain was found to be phylogenetically closely related to Aspergillus niger. Kinetic constants such as K m and V max for extracellular xylanase were determined using various substrate such as beech wood xylan, oat spelt xylan and CM cellulose through Lineweaver-Burk plot. K m , V max and K cat for beech wood xylan are found to be 2.89mg/ml, 2442U and 426178Umlmg -1 respectively. Crude enzyme did not show also CM cellulose activity. The relative efficiency of oat spelt xylan was found to be 0.819 with respect to beech wood xylan. After acid hydrolysis, enzyme was able to produce reducing sugar with 17.7, 35.5, 50.8 and 65% (w/w) from orange peel after 15, 30, 45 and 60min incubation with cellulase free xylanase and maximum reducing sugar formation rate was found to be 55.96μg/ml/min. Therefore, the Aspergillus niger (KP874102.1) is considered as a potential candidate for enzymatic hydrolysis of orange peel. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Xylanase and cellulase activities during anaerobic decomposition of three aquatic macrophytes.

    Science.gov (United States)

    Nunes, Maíra F; da Cunha-Santino, Marcela B; Bianchini, Irineu

    2011-01-01

    Enzymatic activity during decomposition is extremely important to hydrolyze molecules that are assimilated by microorganisms. During aquatic macrophytes decomposition, enzymes act mainly in the breakdown of lignocellulolytic matrix fibers (i.e. cellulose, hemicellulose and lignin) that encompass the refractory fraction from organic matter. Considering the importance of enzymatic activities role in decomposition processes, this study aimed to describe the temporal changes of xylanase and cellulose activities during anaerobic decomposition of Ricciocarpus natans (freely-floating), Oxycaryum cubense (emergent) and Cabomba furcata (submersed). The aquatic macrophytes were collected in Óleo Lagoon, Luiz Antonio, São Paulo, Brazil and bioassays were accomplished.  Decomposition chambers from each species (n = 10) were set up with dried macrophyte fragments and filtered Óleo Lagoon water. The chambers were incubated at 22.5°C, in the dark and under anaerobic conditions. Enzymatic activities and remaining organic matter were measured periodically during 90 days. The temporal variation of enzymes showed that C. furcata presented the highest decay and the highest maximum enzyme production. Xylanase production was higher than cellulase production for the decomposition of the three aquatic macrophytes species.

  1. Amenability of Acacia and Eucalyptus Hardwood Pulps to Elemental Chlorine-Free Bleaching: Application and Efficacy of Microbial Xylanase

    Directory of Open Access Journals (Sweden)

    Avdhesh Kumar Gangwar

    2015-10-01

    Full Text Available This study outlines the results of a biobleaching study of acacia (A. mangium and eucalyptus (E. globulus hardwood kraft pulps with commercial xylanase (Optimase CX 72 L. The comparative study was carried out using an elemental chlorine-free (ECF bleaching sequence (D0EPD1D2 after the enzyme (X stage. The enzyme treatment resulted in improved optical properties with a reduction in bleach chemical consumption. At an equivalent bleach chemical consumption, a brightness gain of 2.1 and 1.7 units and a whiteness gain of 2.7 and 2.3 units were observed with xylanase treatment in acacia and eucalyptus pulps, respectively. In ECF bleaching using the D0EPD1D2 sequence, a final brightness was achieved to the extent of 90% ISO and 89% ISO for acacia and eucalyptus, respectively, at an equivalent charge of bleach chemicals. The post-color (PC number was also reduced by up to 45% for both hardwood pulps compared with the control. The bleachability of acacia was observed to be significantly higher than that of eucalyptus. In addition, a 17.0% and 23.0% reduction in chlorine dioxide and sodium hydroxide, respectively, were obtained for both hardwood pulps after xylanase pre-bleaching, thus indicating an environmentally friendly approach to the process.

  2. An approach to industrial application: influence of black liquor and pH on xylanase efficiency in bleaching of eucalyptus kraft pulp

    OpenAIRE

    Fillat Latorre, Úrsula; Roncero Vivero, María Blanca; Bassa, Alexandre; Sacón, Vera Maria

    2010-01-01

    To obtain a more realistic appraisal of the potential efficiency of xylanases in the industrial bleaching, the influence of pH and the presence of black liquor (measured as COD) on the bleaching efficiency of two commercial xylanases was studied at high temperature. These pH’s, CODs, and temperatures are close to those used in the storage tower of the B fiber line in Jacareı´ unit of Fibria (Brazil). The pulp samples obtained after each bleaching stage were analyzed for kappa number, brigh...

  3. STUDIES ON XYLANASE AND LACCASE ENZYMATIC PREBLEACHING TO REDUCE CHLORINE-BASED CHEMICALS DURING CEH AND ECF BLEACHING

    Directory of Open Access Journals (Sweden)

    Vasanta V. Thakur,

    2012-02-01

    Full Text Available The biobleaching efficiency of xylanase and laccase enzymes was studied on kraft pulps from wood and nonwood based raw materials employed in the Indian paper industry. Treatment of these pulps with xylanase enzyme could result in improved properties, showing 2.0% ISO gain in pulp brightness and/or reducing the demand of chlorine-based bleach chemicals by up to 15% with simultaneous reduction of 20 to 25% in AOX generation in bleach effluents. Further, mill-scale trial results revealed that enzymatic prebleaching can be successfully employed with xylanases to reach the same bleach boosting efficacy. Laccase bleaching was also studied on hardwood pulp at a pH around 8.0, where most of the pulp mills in India are operating, in contrast to earlier studies on laccase enzyme bleaching, which were conducted at acidic pHs, i.e. 4.0 to 5.0. In case of laccase bleaching, interesting results were found wherein a bleach-boosting effect was observed even at pH 8.0. Further studies carried out with HOBT as mediator in comparison to the commonly used and expensive ABTS laccase mediator system (LMS resulted in improvement of the bleaching efficiency with reduction in demand of chlorine dioxide by more than 35%. Potential for further reduction was indicated by the brightness gain, when compared with a control using the DE(pD bleach sequence.

  4. Structural Analysis of Xylanase from Marine Thermophilic Geobacillus stearothermophilus in Tanjung Api, Poso, Indonesia

    Directory of Open Access Journals (Sweden)

    BUDI SAKSONO

    2010-12-01

    Full Text Available A xylanase gene, xynA, has been cloned from thermophilic strain Geobacillus stearothermophilus, which was isolated from marine Tanjung Api, Indonesia. The polymerase chain reaction product of 1266 bp of xynA gene consisted of 1221 bp open reading frame and encoded 407 amino acids including 30 residues of signal peptide. The sequence exhibited highest identity of 98.7% in the level of amino acid, with an extracellular endo-1,4-â-xylanase from G stearothermophilus T-6 (E-GSX T-6 of the glycoside hydrolase family 10 (GH10. A comparative study between the local strain G. stearothermophilus (GSX L and E-GSX T-6 on homology of amino acid sequence indicated five differents amino acids in the gene. They were Threonine/Alanine (T/A, Asparagine/Aspartate (N/D, Lysine/Asparagine (K/N, Isoleucine/Methionine (I/M, Serine/Threonine (S/T at the position 220, 227, 228, 233, and 245, respectively. Protein structural analysis of those differences suggested that those amino acids may play role in biochemical properties such as enzyme stability, in particular its thermostability.

  5. Parthenium sp. as a plant biomass for the production of alkalitolerant xylanase from mutant Penicillium oxalicum SAU{sub E}-3.510 in submerged fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Dwivedi, Pallavi; Vivekanand, V.; Ganguly, Ruma; Singh, Rajesh P. [Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667 (India)

    2009-04-15

    The use of congress grass (Parthenium sp.) and water hyacinth (Eichhornia crassipes) as low cost raw materials for xylanase production from mutant Penicillium oxalicum SAU{sub E}-3.510 in submerged fermentation was investigated. For development of mutant from wild type P. oxalicum SA-8 ITCC 6024, a strategy of mixed mutagenesis was followed using UV-irradiation and ethidium bromide, which had resulted into 1.87 fold increases in the activity of the enzyme. For enzyme production, the fungus was cultivated in mineral medium containing congress grass as carbon source. Considerably higher levels of production (475.2 {+-} 6.0 IU ml{sup -1}) were achieved in media containing congress grass, although it was slightly less than that was obtained (488.5 {+-} 6.5 IU ml{sup -1}) in presence of commercial oat spelt xylan. This fact confirms the feasibility of using this low cost non-food resource as an alternative carbon source to save costs of the enzyme production process. Maximum xylanase activity was reported at 55 C with its stability at 80 C for 2 h. The highest activity of xylanase at pH 9.0 and its stability at similar pH for 24 h denote the alkalitolerant nature of enzyme. (author)

  6. Effects of Xylanase Supplementation on Growth Performance, Nutrient Digestibility and Non-starch Polysaccharide Degradation in Different Sections of the Gastrointestinal Tract of Broilers Fed Wheat-based Diets

    Directory of Open Access Journals (Sweden)

    L. Zhang

    2014-06-01

    Full Text Available This experiment was performed to investigate the effects of exogenous xylanase supplementation on performance, nutrient digestibility and the degradation of non-starch polysaccharides (NSP in different sections of the gastrointestinal tract (GIT of broilers fed wheat-based diets. A total of 120 7-day-old Arbor Acres broiler chicks were randomly allotted to two wheat-based experimental diets supplemented with 0 or 1.0 g/kg xylanase. Each treatment was composed of 6 replicates with 10 birds each. Diets were given to the birds from 7 to 21 days of age. The results showed that xylanase supplementation did not affect feed intake, but increased body weight gain of broiler at 21 day of age by 5.8% (pjejunum>duodenum>>gizzard> caecum. The supplementation of xylanse increased ileal isomaltriose concentration (p<0.05, but did not affect the concentrations of isomaltose, panose and 1-kestose in the digesta of all GIT sections. These results suggest that supplementation of xylanase to wheat-based diets cuts the arabinoxylan backbone into small fragments (mainly arabinose and xylose in the ileum, jejunum and duodenum, and enhances digestibilites of nutrients by decreasing digesta viscosity. The release of arabinose and xylose in the small intestine may also be the important contributors to the growth-promoting effect of xylanase in broilers fed wheat-based diets.

  7. Effects of exogenous phytase and xylanase, individually or in combination, and pelleting on nutrient digestibility, available energy content of wheat and performance of growing pigs fed wheat-based diets.

    Science.gov (United States)

    Yang, Y Y; Fan, Y F; Cao, Y H; Guo, P P; Dong, B; Ma, Y X

    2017-01-01

    Two experiments were conducted to determine the effects of adding exogenous phytase and xylanase, individually or in combination, as well as pelleting on nutrient digestibility, available energy content of wheat and the performance of growing pigs fed wheat-based diets. In Experiment 1, forty-eight barrows with an initial body weight of 35.9±0.6 kg were randomly assigned to a 2×4 factorial experiment with the main effects being feed form (pellet vs meal) and enzyme supplementation (none, 10,000 U/kg phytase, 4,000 U/kg xylanase or 10,000 U/kg phytase plus 4,000 U/kg xylanase). The basal diet contained 97.8% wheat. Pigs were placed in metabolic cages for a 7-d adaptation period followed by a 5-d total collection of feces and urine. Nutrient digestibility and available energy content were determined. Experiment 2 was conducted to evaluate the effects of pelleting and enzymes on performance of wheat for growing pigs. In this experiment, 180 growing pigs (35.2±9.0 kg BW) were allocated to 1 of 6 treatments according to a 2×3 factorial treatment arrangement with the main effects being feed form (meal vs pellet) and enzyme supplementation (0, 2,500 or 5,000 U/kg xylanase). In Experiment 1, there were no interactions between feed form and enzyme supplementation. Pelleting reduced the digestibility of acid detergent fiber (ADF) by 6.4 percentage units (pdigestibility of energy by 0.6 percentage units (pdigestibility of crude protein by 0.5 percentage units (p = 0.07) compared with diets in mash form. The addition of phytase improved the digestibility of phosphorus (pdigestibility of crude protein by 1.0 percentage units (p = 0.09) and increased the digestibility of neutral detergent fiber (NDF) (pdigestibility of phosphorus (pdigestibility (pdigestibility but decreased ADF digestibility. Adding xylanase increased crude protein digestibility and pig performance. Phytase increased the apparent total tract digestibility of phosphorus and calcium. The combination of

  8. Over expression of beta-1, 4-xylanase by auto-induction in E. coli

    International Nuclear Information System (INIS)

    Khan, M.I.K.; Sajjad, M.; Akhtar, W.

    2013-01-01

    Catalytic domain of β-1, 4-xylanase gene, (xynZ.CD) of Clostridium thermocellum was cloned in pET28a expression vector and over-expressed in Escherichia colt BL21 CodonPlus (RIL). The production of XynZ.CD in E. colt was optimized using different concentrations of lactose and induction of the enzyme at different stages of growth. The maximum growth of the cells and the enzyme activity were observed when the cells were induced with 10mM lactose after 8 hours of incubation. The enzyme was found to constitute >40% of the total cell proteins in the supernatant of the lysed cells transformed with recombinant pET28a/xynZ.CD. It was purified by heating the cell lysate at 65 degree C for 30 m followed by fractionation through FPLC. Molecular weight of XynZ.CD was found to be approximately 38,524 D by MALDI-TOF analysis. The enzyme variant was quite stable within broad pH range of 5.5 - 8.0 and it retained >85% of xylanase activity after 2 h incubation at 70 degre C. (author)

  9. Crystallization and preliminary X-ray study of a family 10 alkali-thermostable xylanase from alkalophilic Bacillus sp. strain NG-27

    Energy Technology Data Exchange (ETDEWEB)

    Manikandan, K. [Department of Physics, Indian Institute of Science, Bangalore 560 012 (India); Bhardwaj, Amit [International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067 (India); Ghosh, Amit [Institute of Microbial Technology, Sector 39-A, Chandigarh 160 036 (India); Reddy, V. S. [International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067 (India); Ramakumar, S., E-mail: ramak@physics.iisc.ernet.in [Department of Physics, Indian Institute of Science, Bangalore 560 012 (India); Bioinformatics Centre, Indian Institute of Science, Bangalore 560 012 (India)

    2005-08-01

    A family 10 alkali-thermostable xylanase from Bacillus sp. NG-27 has been crystallized. A diffraction data set has been collected to 2.2 Å resolution. Xylanases (EC 3.2.1.8) catalyze the hydrolysis of β-1,4-glycosidic linkages within xylan, a major hemicellulose component in the biosphere. The extracellular endoxylanase (XylnA) from the alkalophilic Bacillus sp. strain NG-27 belongs to family 10 of the glycoside hydrolases. It is active at 343 K and pH 8.4. Moreover, it has attractive features from the point of view of utilization in the paper pulp, animal feed and baking industries since it is an alkali-thermostable protein. In this study, XylnA was purified from the native host source and crystallized by the hanging-drop vapour-diffusion method. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 174.5, b = 54.7, c = 131.5 Å, β = 131.2°, and diffract to better than 2.2 Å resolution.

  10. Crystallization and preliminary X-ray study of a family 10 alkali-thermostable xylanase from alkalophilic Bacillus sp. strain NG-27

    International Nuclear Information System (INIS)

    Manikandan, K.; Bhardwaj, Amit; Ghosh, Amit; Reddy, V. S.; Ramakumar, S.

    2005-01-01

    A family 10 alkali-thermostable xylanase from Bacillus sp. NG-27 has been crystallized. A diffraction data set has been collected to 2.2 Å resolution. Xylanases (EC 3.2.1.8) catalyze the hydrolysis of β-1,4-glycosidic linkages within xylan, a major hemicellulose component in the biosphere. The extracellular endoxylanase (XylnA) from the alkalophilic Bacillus sp. strain NG-27 belongs to family 10 of the glycoside hydrolases. It is active at 343 K and pH 8.4. Moreover, it has attractive features from the point of view of utilization in the paper pulp, animal feed and baking industries since it is an alkali-thermostable protein. In this study, XylnA was purified from the native host source and crystallized by the hanging-drop vapour-diffusion method. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 174.5, b = 54.7, c = 131.5 Å, β = 131.2°, and diffract to better than 2.2 Å resolution

  11. Cellulase and xylanase activity during the decomposition of three aquatic macrophytes in a tropical oxbow lagoon

    Directory of Open Access Journals (Sweden)

    L Sciessere

    2011-09-01

    Full Text Available Due to the connection between enzymatic activity and degradation of different fractions of organic matter, enzyme assays can be used to estimate degradation rates of particulate and dissolved organic carbon in freshwater systems. The aim of this study was to quantify and model the enzymatic degradation involving the decomposition of macrophytes, describing temporal activity of cellulases (EC 3.2.1.4 and EC 3.2.1.91 and xylanase (EC 3.2.1.8 during in situ decomposition of three aquatic macrophytes (Salvinia sp., Eichhornia azurea and Cyperus giganteus on the surface and water-sediment interface (w-s interface of an oxbow lagoon (Óleo lagoon within a natural Brazilian Savanna Reserve. Overall, the enzymatic degradation of aquatic macrophytes in Óleo lagoon occurred during the whole year and was initiated together with leaching. Xylanase production was ca. 5 times higher than cellulase values due to easy access to this compound by cellulolytic microorganisms. Enzymatic production and detritus mass decay were similar on the surface and w-s interface. Salvinia sp. was the most recalcitrant detritus, with low mass decay and enzymatic activity. E. azurea and C. giganteus decomposition rates and enzymatic production were high and similar. Due to the physicochemical homogeneity observed in the Óleo lagoon, the differences between the decay rates of each species are mostly related with detritus chemical quality.

  12. Scan-rate dependence in protein calorimetry: the reversible transitions of Bacillus circulans xylanase and a disulfide-bridge mutant.

    OpenAIRE

    Davoodi, J.; Wakarchuk, W. W.; Surewicz, W. K.; Carey, P. R.

    1998-01-01

    The stabilities of Bacillus circulans xylanase and a disulfide-bridge-containing mutant (S100C/N148C) were investigated by differential scanning calorimetry (DSC) and thermal inactivation kinetics. The thermal denaturation of both proteins was found to be irreversible, and the apparent transition temperatures showed a considerable dependence upon scanning rate. In the presence of low (nondenaturing) concentrations of urea, calorimetric transitions were observed for both proteins in the second...

  13. Single-step purification and characterization of an extreme halophilic, ethanol tolerant and acidophilic xylanase from Aureobasidium pullulans NRRL Y-2311-1 with application potential in the food industry.

    Science.gov (United States)

    Yegin, Sirma

    2017-04-15

    An extracellular xylanase from Aureobasidium pullulans NRRL Y-2311-1 produced on wheat bran was purified by a single-step chromatographic procedure. The enzyme had a molecular weight of 21.6kDa. The optimum pH and temperature for xylanase activity were 4.0 and 30-50°C, respectively. The enzyme was stable in the pH range of 3.0-8.0. The inactivation energy of the enzyme was calculated as 218kJmol -1 . The xylanase was ethanol tolerant and kept complete activity in the presence of 10% ethanol. Likewise, it retained almost complete activity at a concentration range of 0-20% NaCl. In general, the enzyme was resistant to several metal ions and reagents. Mg 2+ , Zn 2+ , Cu 2+ , K 1+ , EDTA and β-mercaptoethanol resulted in enhanced xylanase activity. The K m and V max values on beechwood xylan were determined to be 19.43mgml -1 and 848.4Uml -1 , respectively. The enzyme exhibits excellent characteristics and could, therefore, be a promising candidate for application in food and bio-industries. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Engineering increased thermostability in the GH-10 endo-1,4-ß-xylanase from Thermoascus aurantiacus CBMAI 756

    Science.gov (United States)

    The GH10 endo-xylanase from Thermoascus aurantiacus CBMAI 756 (XynA) is industrially attractive due to its considerable thermostability and high specific activity. Considering the possibility of a further improvement in thermostability, eleven mutants were created in the present study via site-direc...

  15. Xylanase and feruloyl esterase from actinomycetes cultures could enhance sugarcane bagasse hydrolysis in the production of fermentable sugars.

    Science.gov (United States)

    Rahmani, Nanik; Kahar, Prihardi; Lisdiyanti, Puspita; Hermiati, Euis; Lee, Jaemin; Yopi; Prasetya, Bambang; Ogino, Chiaki; Kondo, Akihiko

    2018-02-23

    The addition of enzymes that are capable of degrading hemicellulose has a potential to reduce the need for commercial enzymes during biomass hydrolysis in the production of fermentable sugars. In this study, a high xylanase producing actinomycete strain (Kitasatospora sp. ID06-480) and the first ethyl ferulate producing actinomycete strain (Nonomuraea sp. ID06-094) were selected from 797 rare actinomycetes, respectively, which were isolated in Indonesia. The addition (30%, v/v) of a crude enzyme supernatant from the selected strains in sugarcane bagasse hydrolysis with low-level loading (1 FPU/g-biomass) of Cellic® CTec2 enhanced both the released amount of glucose and reducing sugars. When the reaction with Ctec2 was combined with crude enzymes containing either xylanase or feruloyl esterase, high conversion yield of glucose from cellulose at 60.5% could be achieved after 72 h-saccharification.

  16. Prediction of ingredient quality and the effect of a combination of xylanase, amylase, protease and phytase in the diets of broiler chicks. 2. Energy and nutrient utilisation.

    Science.gov (United States)

    Cowieson, A J; Singh, D N; Adeola, O

    2006-08-01

    1. In order to investigate the effects of xylanase, amylase, protease and phytase in the diets of broiler chickens containing graded concentrations of metabolisable energy (ME), two 42-d experiments were conducted using a total of 2208 broiler chicks (8 treatments with 12 replicate pens in each experiment). 2. Four diets including one positive and three negative control diets were used. Three maize/soybean meal-based negative control (NC) diets were formulated to be identical in available phosphorus (P), calcium (Ca) and amino acids but NC1 contained approximately 0.17 MJ/kg less ME than NC2 and approximately 0.34 MJ/kg less ME than NC3. A positive control (PC) was fed for comparison and was formulated to be adequate in all nutrients, providing approximately 0.63 MJ/kg ME, 0.13% available P, 0.12% Ca and 1 to 2% amino acids more than NC1. 3. The reduction in nutrient density between NC1 and PC was determined using ingredient quality models Avichecktrade mark Corn and Phychecktrade mark that can predict the response to exogenous enzymes in maize/soybean meal-based broiler diets. Supplementation of each diet with or without a cocktail of xylanase, amylase, protease and phytase gave a total of 8 dietary treatments in a 4 x 2 factorial arrangement. The same treatments and diet designs were used in both experiments but conducted in different locations using different batches of maize, soybean meal and minor ingredients. 4. In both experiments, digestibility was improved by the addition of exogenous enzymes, particularly those for P, Ca and certain amino acids. In addition, the supplementation of the PC with enzymes elicited a positive response indicating that over-the-top addition of xylanase, amylase, protease and phytase may offer a nutritionally and economically viable alternative to feed cost reduction. 5. It can be concluded that the digestibility of nutrients by broilers fed on maize/soybean meal-based diets can be improved by the use of a combination of xylanase

  17. Efficient production of lignocellulolytic enzymes xylanase, β-xylosidase, ferulic acid esterase and β-glucosidase by the mutant strain Aspergillus awamori 2B.361 U2/1

    Directory of Open Access Journals (Sweden)

    Leda Maria Fortes Gottschalk

    2013-01-01

    Full Text Available The production of xylanase, β-xylosidase, ferulic acid esterase and β-glucosidase by Aspergillus awamori 2B.361 U2/1, a hyper producer of glucoamylase and pectinase, was evaluated using selected conditions regarding nitrogen nutrition. Submerged cultivations were carried out at 30 ºC and 200 rpm in growth media containing 30 g wheat bran/L as main carbon source and either yeast extract, ammonium sulfate, sodium nitrate or urea, as nitrogen sources; in all cases it was used a fixed molar carbon to molar nitrogen concentration of 10.3. The use of poor nitrogen sources favored the accumulation of xylanase, β-xylosidase and ferulic acid esterase to a peak concentrations of 44,880; 640 and 118 U/L, respectively, for sodium nitrate and of 34,580, 685 and 170 U/L, respectively, for urea. However, the highest β-glucosidase accumulation of 10,470 U/L was observed when the rich organic nitrogen source yeast extract was used. The maxima accumulation of filter paper activity, xylanase, β-xylosidase, ferulic acid esterase and β-glucosidase by A. awamori 2B.361 U2/1 was compared to that produced by Trichoderma reesei Rut-C30. The level of β-glucosidase was over 17-fold higher for the Aspergillus strain, whereas the levels of xylanase and β-xylosidase were over 2-fold higher. This strain also produced ferulic acid esterase (170 U/L, which was not detected in the T. reesei culture.

  18. A high performance Trichoderma reesei strain that reveals the importance of xylanase III in cellulosic biomass conversion.

    Science.gov (United States)

    Nakazawa, Hikaru; Kawai, Tetsushi; Ida, Noriko; Shida, Yosuke; Shioya, Kouki; Kobayashi, Yoshinori; Okada, Hirofumi; Tani, Shuji; Sumitani, Jun-Ichi; Kawaguchi, Takashi; Morikawa, Yasushi; Ogasawara, Wataru

    2016-01-01

    The ability of the Trichoderma reesei X3AB1strain enzyme preparations to convert cellulosic biomass into fermentable sugars is enhanced by the replacement of xyn3 by Aspergillus aculeatus β-glucosidase 1 gene (aabg1), as shown in our previous study. However, subsequent experiments using T. reesei extracts supplemented with the glycoside hydrolase (GH) family 10 xylanase III (XYN III) and GH Family 11 XYN II showed increased conversion of alkaline treated cellulosic biomass, which is rich in xylan, underscoring the importance of XYN III. To attain optimal saccharifying potential in T. reesei, we constructed two new strains, C1AB1 and E1AB1, in which aabg1 was expressed heterologously by means of the cbh1 or egl1 promoters, respectively, so that the endogenous XYN III synthesis remained intact. Due to the presence of wild-type xyn3 in T. reesei E1AB1, enzymes prepared from this strain were 20-30% more effective in the saccharification of alkaline-pretreated rice straw than enzyme extracts from X3AB1, and also outperformed recent commercial cellulase preparations. Our results demonstrate the importance of XYN III in the conversion of alkaline-pretreated cellulosic biomass by T. reesei. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. Isothermal titration calorimetry and surface plasmon resonance allow quantifying substrate binding to different binding sites of Bacillus subtilis xylanase

    DEFF Research Database (Denmark)

    Cuyvers, Sven; Dornez, Emmie; Abou Hachem, Maher

    2012-01-01

    Isothermal titration calorimetry and surface plasmon resonance were tested for their ability to study substrate binding to the active site (AS) and to the secondary binding site (SBS) of Bacillus subtilis xylanase A separately. To this end, three enzyme variants were compared. The first...

  20. Finding stable cellulase and xylanase: evaluation of the synergistic effect of pH and temperature.

    Science.gov (United States)

    Farinas, Cristiane S; Loyo, Marcel Moitas; Baraldo, Anderson; Tardioli, Paulo W; Neto, Victor Bertucci; Couri, Sonia

    2010-12-31

    Ethanol from lignocellulosic biomass has been recognized as one of the most promising alternatives for the production of renewable and sustainable energy. However, one of the major bottlenecks holding back its commercialization is the high costs of the enzymes needed for biomass conversion. In this work, we studied the enzymes produced from a selected strain of Aspergillus niger under solid state fermentation. The cellulase and xylanase enzymatic cocktail was characterized in terms of pH and temperature by using response surface methodology. Thermostability and kinetic parameters were also determined. The statistical analysis of pH and temperature effects on enzymatic activity showed a synergistic interaction of these two variables, thus enabling to find a pH and temperature range in which the enzymes have a higher activity. The results obtained allowed the construction of mathematical models used to predict endoglucanase, β-glucosidase and xylanase activities under different pH and temperature conditions. Optimum temperature values for all three enzymes were found to be in the range between 35°C and 60°C, and the optimum pH range was found between 4 and 5.5. The methodology employed here was very effective in estimating enzyme behavior under different process conditions. Copyright © 2010 Elsevier B.V. All rights reserved.

  1. Influence of organic nitrogen amendment, containing amino acids on the cellulase and xylanase, produced by Trichoderma spp. isolates

    Directory of Open Access Journals (Sweden)

    D. Draganova

    2017-09-01

    Full Text Available Abstract. Cellulases and hemicellulases are amount the main hydrolytic enzymes, involved in the bioconversion of lignocellulose material by microorganisms. Filamentous fungi of the genus Trichoderma are one of the most studied and good producer of cellulases and hemicellulases. The nutrients balance, especially carbon to nitrogen ratio, is one of the main factors of the biodegradation. The ability of 37 local isolates of Trichoderma sp. to produce cellulases and xylanase were tested in solid state cultivation on wheat straw as a substrate whit two variants: 1. the straw was only moistured with destilated water (CN 80:1; 2. the C:N ratio of the straw was adjusted to 30:1 using organic nitrogen amendment. There is a significant difference in the enzymatic activity of the isolates in their cultivation on straw with CN 80 and CN 30. The highest carboxymethylcellulase (CMCase activity at CN 80 showed T1T (110.19U/ml, and in the variant at CN 30 - TD (369.07U/ml. The highest β-glucosidase activity on both variants CN 80 and CN 30 was established for TG (2743.1U/ml - 12679.9U/ml. The highest xylanase activity at CN 80 and CN 30 was measured on T4I (21311.5U/ml – 47937.5U/ml. After ONA addition, all enzymes activities have increased several times, indicating the enhancing effect of the additive. The average activity of CMCase increased 6.1 times, the average β - glucosidase activity increased 5.1 times, while the xylanase activity increased 4.9 times for all tested isolates. The increase in activity of the investigated enzymes showed different patterns.

  2. Xylanase Increased the Ileal Digestibility of Non-Starch Polysaccharides and Concentration of Low Molecular Weight Non-Digestible Carbohydrates in Pigs Fed High Levels of wheat DDGS

    DEFF Research Database (Denmark)

    Pedersen, Mads Brøgger; Yu, Shukun; Arent, Susan

    2015-01-01

    The objective was to study the effect of a commercially available xylanase (CAX), an experimental xylanase (EX), and EX in combination with protease (EXP) on the degradation of nondigestible carbohydrates (NDC) and apparent ileal digestibility (AID) of nutrients in wheat distillers dried grains...... with solubles (wDDGS). The control and 3 enzyme diets contained 96% wDDGS supplemented with vitamins, minerals, l-lysine, and chromic oxide as a digestibility marker in addition to enzyme premix. Eight ileal cannulated pigs were fed 4 experimental diets containing 96% wDDGS—a control diet or 1 of 3 diets...

  3. Supplementation with xylanase and β-xylosidase to reduce xylo-oligomer and xylan inhibition of enzymatic hydrolysis of cellulose and pretreated corn stover

    Science.gov (United States)

    2011-01-01

    Background Hemicellulose is often credited with being one of the important physical barriers to enzymatic hydrolysis of cellulose, and acts by blocking enzyme access to the cellulose surface. In addition, our recent research has suggested that hemicelluloses, particularly in the form of xylan and its oligomers, can more strongly inhibit cellulase activity than do glucose and cellobiose. Removal of hemicelluloses or elimination of their negative effects can therefore become especially pivotal to achieving higher cellulose conversion with lower enzyme doses. Results In this study, cellulase was supplemented with xylanase and β-xylosidase to boost conversion of both cellulose and hemicellulose in pretreated biomass through conversion of xylan and xylo-oligomers to the less inhibitory xylose. Although addition of xylanase and β-xylosidase did not necessarily enhance Avicel hydrolysis, glucan conversions increased by 27% and 8% for corn stover pretreated with ammonia fiber expansion (AFEX) and dilute acid, respectively. In addition, adding hemicellulase several hours before adding cellulase was more beneficial than later addition, possibly as a result of a higher adsorption affinity of cellulase and xylanase to xylan than glucan. Conclusions This key finding elucidates a possible mechanism for cellulase inhibition by xylan and xylo-oligomers and emphasizes the need to optimize the enzyme formulation for each pretreated substrate. More research is needed to identify advanced enzyme systems designed to hydrolyze different substrates with maximum overall enzyme efficacy. PMID:21702938

  4. Evaluation of operational parameters on the precipitation of endoglucanase and xylanase produced by solid state fermentation of Aspergillus niger

    Directory of Open Access Journals (Sweden)

    C. S. Farinas

    2011-03-01

    Full Text Available In order to develop cost effective processes for converting biomass into biofuels, it is essential to improve enzyme production yields, stability and specific activity. In this context, the aim of this work was to evaluate the concentration of two enzymes involved in the hydrolysis of biomass, endoglucanase and xylanase, through precipitation. Statistical experimental design was used to evaluate the influence of precipitant agent concentration (ammonium sulfate and ethanol, aging time, and temperature on enzyme activity recovery. Precipitant agent concentration and aging time showed a statistically significant effect at the 95% confidence level, on both enzyme activity recoveries. The recovery of endoglucanase with ammonium sulfate and ethanol reached values up to 65 and 61%, respectively. For xylanase, the recovery rates were lower, 27 and 25% with ammonium sulfate and ethanol, respectively. The results obtained allowed the selection of the variables relevant to improving enzyme activity recovery within operational conditions suitable for industrial applications.

  5. Isolation, purification and characterization of xylanase produced by Arthrobacter sp. MTCC 5214 when grown in solid-state fermentation

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Bhosle, N.B.

    %) fractionation, and purified to homogeneity using size exclusion and ion exchange chromatography. The molecular mass of xylanase was approx. 20 kDa. Enzyme retained 100% activity at pH 7 and 8 for 24 h. It was interesting to note that at higher pH such as 9, 10...

  6. Heterologous expression, purification, crystallization and preliminary X-ray analysis of Trichoderma reesei xylanase II and four variants

    International Nuclear Information System (INIS)

    Wan, Qun; Kovalevsky, Andrey; Zhang, Qiu; Hamilton-Brehm, Scott; Upton, Rosalynd; Weiss, Kevin L.; Mustyakimov, Marat; Graham, David; Coates, Leighton; Langan, Paul

    2013-01-01

    The wild-type protein and four active-site mutants of xylanase II from Trichoderma reesei that catalyzes the hydrolysis of glycosidic bonds in xylan have successfully been crystallized. The crystallization of several structures including ligand-free and protein ligand complexes containing the substrate (xylohexaose) or product (xylotriose) are detailed. Xylanase II from Trichoderma reesei catalyzes the hydrolysis of glycosidic bonds in xylan. Crystallographic studies of this commercially important enzyme have been initiated to investigate its reaction mechanism, substrate binding and dependence on basic pH conditions. The wild-type protein was heterologously expressed in an Escherichia coli host using the defined medium and four active-site amino acids were replaced to abolish its activity (E177Q and E86Q) or to change its pH optimum (N44D and N44H). Cation-exchange and size-exclusion chromatography were used to obtain >90% protein purity. The ligand-free proteins and variant complexes containing substrate (xylohexaose) or product (xylotriose) were crystallized in several different space groups and diffracted to high resolutions (from 1.07 to 1.55 Å)

  7. Cellulase and Xylanase Production by Penicillium echinulatum in Submerged Media Containing Cellulose Amended with Sorbitol

    Directory of Open Access Journals (Sweden)

    Carla Eliana Todero Ritter

    2013-01-01

    Full Text Available The present work investigated the use of sorbitol as a soluble carbon source, in association with cellulose, to produce cellulases and xylanases in submerged cultures of Penicillium echinulatum 9A02S1. Because cellulose is an insoluble carbon source, in cellulase production, there are some problems with rheology and oxygen transfer. The submerged fermentations containing media composed of 0, 0.25, 0.5, 0.75, and 1% (w/v sorbitol and cellulose that were added at different times during the cultivation; 0.2% (w/v soy bran; 0.1% (w/v wheat bran; and a solution of salts. The highest filter paper activity (FPA ( IU·mL−1 was obtained on the seventh day in the medium containing 0.5% (w/v sorbitol and 0.5% (w/v cellulose added 24 h after the start of cultivation. However, the CMCases showed an activity peak on the sixth day ( IU·mL−1 in the medium containing 0.75% (w/v sorbitol and 0.75% (w/v cellulose added after 12 h of cultivation. The xylanases showed the highest activity in the medium with 0.75% (w/v sorbitol and 0.25% (w/v cellulose added 36 h after the start of cultivation. This strategy enables the reduction of the cellulose concentration, which in high concentrations can cause rheological and oxygen transfer problems.

  8. Supplementation with xylanase and β-xylosidase to reduce xylo-oligomer and xylan inhibition of enzymatic hydrolysis of cellulose and pretreated corn stover

    Directory of Open Access Journals (Sweden)

    Qing Qing

    2011-06-01

    Full Text Available Abstract Background Hemicellulose is often credited with being one of the important physical barriers to enzymatic hydrolysis of cellulose, and acts by blocking enzyme access to the cellulose surface. In addition, our recent research has suggested that hemicelluloses, particularly in the form of xylan and its oligomers, can more strongly inhibit cellulase activity than do glucose and cellobiose. Removal of hemicelluloses or elimination of their negative effects can therefore become especially pivotal to achieving higher cellulose conversion with lower enzyme doses. Results In this study, cellulase was supplemented with xylanase and β-xylosidase to boost conversion of both cellulose and hemicellulose in pretreated biomass through conversion of xylan and xylo-oligomers to the less inhibitory xylose. Although addition of xylanase and β-xylosidase did not necessarily enhance Avicel hydrolysis, glucan conversions increased by 27% and 8% for corn stover pretreated with ammonia fiber expansion (AFEX and dilute acid, respectively. In addition, adding hemicellulase several hours before adding cellulase was more beneficial than later addition, possibly as a result of a higher adsorption affinity of cellulase and xylanase to xylan than glucan. Conclusions This key finding elucidates a possible mechanism for cellulase inhibition by xylan and xylo-oligomers and emphasizes the need to optimize the enzyme formulation for each pretreated substrate. More research is needed to identify advanced enzyme systems designed to hydrolyze different substrates with maximum overall enzyme efficacy.

  9. Purification and characterization of endo-xylanases from Aspergillus Niger. III. An enzyme of PL 365

    Energy Technology Data Exchange (ETDEWEB)

    Fournier, R.A.; Frederick, M.M.; Frederick, J.R.; Reilly, P.J.

    1985-04-01

    An endo-xylanase (1,4-..beta..-D-xylan xylanohydrolase, EC 3.2.1.8) from Aspergillus niger was purified to homogeneity by chromatography with Ultrogel AcA 54, SP-Sephadex C-25 at pH 4.5, DEAE-Sephadex A-25 at pH 5.4, Sephadex G-50, and DEAE-Sephadex A-25 at pH 5.15. The enzyme was active on soluble xylan, on insoluble xylan only after arabinosyl-initiated branch points were removed, and on xylooligosaccharides longer than xylotetraose. There was slight activity on carboxymethyl-cellulose, arabinogalactan, glucomannan, and p-nitrophenyl-..beta..-D- glucopyranoside. The main products of the hydrolysis of soluble and insoluble xylan were oligosaccharides of intermediate length, especially the tri- and pentasaccharides. The isolectric point of the enzyme was 3.65. It had a molecular weight of 2.8 x 10/sup 4/ by SDS-gel electrophoresis, and was high in acidic amino acids but low in those containing sulfur. Highest activity in a 20-min assay at pH 5 was between 40 and 45 degrees C, with an activation energy up to 40 degrees C of 11.1 kJ/mol. The optimum pH for activity was at 5.0. The enzyme was strongly activated by Ca/sup 2 +/. 15 references.

  10. Utilization of Bagasse Cellulose for Ethanol Production through Simultaneous Saccharification and Fermentation by Xylanase

    Directory of Open Access Journals (Sweden)

    M Samsuri

    2010-10-01

    Full Text Available Bagasse is a solid residue from sugar cane process, which is not many use it for some product which have more added value. Bagasse, which is a lignosellulosic material, be able to be use for alternative energy resources like bioethanol or biogas. With renewable energy resources a crisis of energy in Republic of Indonesia could be solved, especially in oil and gas. This research has done the conversion of bagasse to bioethanol with xylanase enzyme. The result show that bagasse contains of 52,7% cellulose, 20% hemicelluloses, and 24,2% lignin. Xylanase enzyme and Saccharomyces cerevisiae was used to hydrolyse and fermentation in SSF process. Variation in this research use pH (4, 4,5, and 5, for increasing ethanol quantity, SSF process was done by added chloride acid (HCl with concentration 0.5% and 1% (v/v and also pre-treatment with white rot fungi such as Lentinus edodes (L.edodes as long 4 weeks. The SSF process was done with 24, 48, 72, and 96 hour's incubation time for fermentation. Variation of pH 4, 4,5, and 5 can produce ethanol with concentrations 2,357 g/L, 2,451 g/L, 2,709 g/L. The added chloride acid (HCl with concentration 0.5% and 1% (v/v and L. edodes can increase ethanol yield, The highest ethanol concentration with added chloride acid (HCl concentration 0.5% and 1% consecutively is 2,967 g/L, 3,249 g/L. The highest ethanol concentration with pre-treatment by L. edodes is 3,202 g/L.

  11. Improving the performance of dairy cattle with a xylanase-rich exogenous enzyme preparation.

    Science.gov (United States)

    Romero, J J; Macias, E G; Ma, Z X; Martins, R M; Staples, C R; Beauchemin, K A; Adesogan, A T

    2016-05-01

    The objective of this experiment was to examine effects of adding 2 exogenous fibrolytic enzymes (EFE) to the total mixed ration (TMR) on the performance of lactating dairy cows (experiment 1) and the kinetics of ruminal degradation of the diet (experiment 2). Twelve EFE had been screened in a series of in vitro assays that identified the most potent EFE and their optimal doses for increasing the digestibility of bermudagrass. In experiment 1, 66 Holstein cows (21±5 d in milk) were grouped by previous milk production and parity (45 multiparous and 21 primiparous) and assigned randomly to 1 of the following 3 treatments: (1) control (CON, untreated), (2) Xylanase Plus [2A, 1mL/kg of TMR dry matter (DM); Dyadic International, Jupiter, FL], and (3) a 75:25 (vol/vol) mixture of Cellulase Plus and Xylanase Plus EFE (3A, 3.4mL/kg of TMR DM; Dyadic International). The EFE were sprayed twice daily onto a TMR (10% bermudagrass silage, 35% corn silage, 5% alfalfa-orchardgrass hay mixture, and 50% concentrates; DM basis) and fed for a 14-d training and covariate period and a 70-d measurement period. Experiment 2 aimed to examine the in situ DM ruminal degradability and ruminal fermentation measurements of the diets fed in experiment 1. Three ruminally fistulated lactating Holstein cows were assigned to the diets. The experiment had a 3×3 Latin square design with 23-d periods. In experiment 1, application of 2A increased intakes (kg/d) of DM (23.5 vs. 22.6), organic matter (21.9 vs. 20.9), and crude protein (3.9 vs. 3.7) and tended to increase yields (kg/d) of fat-corrected milk (41.8 vs. 40.7) and milk fat (1.48 vs. 1.44). In particular, 2A increased milk yield (kg/d) during wk 3 (41.2 vs. 39.8, tendency), 6 (41.9 vs. 40.1), and 7 (42.1 vs. 40.4), whereas 3A increased milk yield (kg/d) during wk 6 (41.5 vs. 40.1, tendency), 8 (41.8 vs. 40.0), and 9 (40.9 vs. 39.5, tendency). In experiment 2, EFE treatment did not affect ruminal DM degradation kinetics or ruminal pH, ammonia

  12. Effects of diet acidification and xylanase supplementation on performance, nutrient digestibility, duodenal histology and gut microflora of broilers fed wheat based diet

    NARCIS (Netherlands)

    Esmaeilipour, O.; Moravej, H.; Shivazad, M.; Rezaian, M.; Aminzadeh, S.; Krimpen, van M.M.

    2012-01-01

    1. The objective of this experiment was to study the influences of xylanase and citric acid on the performance, nutrient digestibility, digesta viscosity, duodenal histology, and gut microflora of broilers fed on a wheat based diet. 2. The experiment was carried out as a 2 x 3 factorial arrangement

  13. Valorization of Brewer's spent grain to prebiotic oligosaccharide: Production, xylanase catalyzed hydrolysis, in-vitro evaluation with probiotic strains and in a batch human fecal fermentation model.

    Science.gov (United States)

    Sajib, Mursalin; Falck, Peter; Sardari, Roya R R; Mathew, Sindhu; Grey, Carl; Karlsson, Eva Nordberg; Adlercreutz, Patrick

    2018-02-20

    Brewer's spent grain (BSG) accounts for around 85% of the solid by-products from beer production. BSG was first extracted to obtain water-soluble arabinoxylan (AX). Using subsequent alkali extraction (0.5 M KOH) it was possible to dissolve additional AX. In total, about 57% of the AX in BSG was extracted with the purity of 45-55%. After comparison of nine xylanases, Pentopan mono BG, a GH11 enzyme, was selected for hydrolysis of the extracts to oligosaccharides with minimal formation of monosaccharides. Growth of Bifidobacterium adolescentis (ATCC 15703) was promoted by the enzymatic hydrolysis to arabinoxylooligosaccharides, while Lactobacillus brevis (DSMZ 1264) utilized only unsubstituted xylooligosaccharides. Furthermore, utilization of the hydrolysates by human gut microbiota was also assessed in a batch human fecal fermentation model. Results revealed that the rates of fermentation of the BSG hydrolysates by human gut microbiota were similar to that of commercial prebiotic fructooligosaccharides, while inulin was fermented at a slower rate. In summary, a sustainable process to valorize BSG to functional food ingredients has been proposed. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. Purification and characterization of endo-xylanases from aspergillus Niger. II. An enzyme of PL 45

    Energy Technology Data Exchange (ETDEWEB)

    Shei, J.C.; Fratzke, A.R.; Frederick, M.M.; Frederick, J.R.; Reilly, P.J.

    1985-04-01

    A homogeneous endo-xylanase (1,4-..beta..-D-xylan xylano-hydrolase, EC 3.2.1.8) was obtained from a crude Aspergillus niger pentosanase by chromatography with Ultrogel AcA 54, SP-Sephadex C-25 at pH 4.5, DEAE-Sephadex A-25 at pH 5.4, Sephadex G-50, and SP-Sephadex C-25 with a gradient from pH 2.8 to pH 4.6. It was much more active on soluble than on insoluble xylan yielding large amounts of unreacted xylan and a mixture of oligosaccharides with chain lengths from two to six. No xylose or L-arabinose was produced. There was high activity on a xylopentaose through xylononaose mixture, but not on xylobiose, xylotriose, or xylotetraose. The enzyme had slight activity on untreated cellulose, carboxymethylcellulose, and pectin. Molecular weight was ca. 1.4 x 10/sup 4/, with an isoelectric point of 4.5 and an amino acid profile high in acidic but low in sulfur-containing residues. In a 25-min assay at pH 4.7, this endo-xylanase was most active at 45 degrees C, with an activation energy from 5 to 35 degrees C of 33.3 kJ/mol. The optimum pH for activity was 4.9. Decay in buffer was first order, with an activation energy at pH 4.7 from 48 to 53 degrees C of 460 kJ/mol. Optimum pH for stability was about 5.6, where the half-life at 48 degrees C in buffer was ca. 40 h.

  15. Biosynthesis, purification and characterization of endoglucanase from a xylanase producing strain Aspergillus niger B03

    Directory of Open Access Journals (Sweden)

    Georgi Todorov Dobrev

    2012-03-01

    Full Text Available An extracellular endoglucanase was isolated from the culture liquid of xylanase producing strain Aspergillus niger B03. The enzyme was purified to a homogenous form, using consecutive ultrafiltration, anion exchange chromatography, and gel filtration. Endoglucanase was a monomer protein with a molecular weight of 26,900 Da determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 28,800 Da determined by gel filtration. The optimal pH and temperature values for the enzyme action were 3.5 and 65ºC respectively. Endoglucanase was stable at 40ºC, pH 3.0 for 210 min. The substrate specificity of the enzyme was determined with carboxymethyl cellulose, filter paper, and different glycosides. Endoglucanase displayed maximum activity in the case of carboxymethyl cellulose, with a Km value of 21.01 mg/mL. The substrate specificity and the pattern of substrate degradation suggested that the enzyme is an endoglucanase. Endoglucanase showed a synergism with endoxylanase in corn cobs hydrolysis.

  16. Purification, crystallization and crystallographic analysis of Clostridium thermocellum endo-1,4-β-d-xylanase 10B in complex with xylohexaose

    Energy Technology Data Exchange (ETDEWEB)

    Najmudin, Shabir, E-mail: shabir@dq.fct.unl.pt [REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica (Portugal); Pinheiro, Benedita A. [CIISA - Faculdade de Medicina Veterinária, Universidade Técnica de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisboa (Portugal); Romão, Maria J. [REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica (Portugal); Prates, José A. M.; Fontes, Carlos M. G. A., E-mail: shabir@dq.fct.unl.pt [CIISA - Faculdade de Medicina Veterinária, Universidade Técnica de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisboa (Portugal); REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica (Portugal)

    2008-08-01

    The N-terminal moiety of C. thermocellum endo-1,4-β-d-xylanase 10B, comprising a carbohydrate-binding module (CBM22-1) and a GH10 E337A mutant domain, has been crystallized in complex with xylohexaose. The crystals belong to the trigonal space group P3{sub 2}21, contain a dimer in the asymmetric unit and diffract to beyond 2.0 Å resolution. The cellulosome of Clostridium thermocellum is a highly organized multi-enzyme complex of cellulases and hemicellulases involved in the hydrolysis of plant cell-wall polysaccharides. The bifunctional multi-modular xylanase Xyn10B is one of the hemicellulase components of the C. thermocellum cellulosome. The enzyme contains an internal glycoside hydrolase family 10 catalytic domain (GH10) and a C-terminal family 1 carbohydrate esterase domain (CE1). The N-terminal moiety of Xyn10B (residues 32–551), comprising a carbohydrate-binding module (CBM22-1) and the GH10 E337A mutant, was crystallized in complex with xylohexaose. The crystals belong to the trigonal space group P3{sub 2}21 and contain a dimer in the asymmetric unit. The crystals diffracted to beyond 2.0 Å resolution.

  17. Fodder radish cake (Raphanus sativus L. as an alternative biomass for the production of cellulases and xylanases in solid-state cultivation

    Directory of Open Access Journals (Sweden)

    L. Zukovski

    Full Text Available Abstract Fodder radish (FR is an oilseed crop with a high potential for biodiesel production due to its high productivity and the quality of its seed oil. FR oil extraction results in a residue that is rich in protein and fiber. In this study, FR cake (FRC was evaluated as carbon and nitrogen source for the production of cellulases and xylanases using Penicillium echinulatum S1M29 during solid-state cultivation. It was determined that it is possible to partially replace wheat bran (WB by FRC, resulting in 24.22 ± 0.25U/g Filter Paper Activity (144 hours, 210.5 ± 5.8U/g endoglucanase activity (144 hours, 22.62 ± 0.01U/g (-glucosidase activity (96 hours and 784.7 ± 70.19U/g xylanase activity (120 hours. These values are equal or higher than the enzymatic activity obtained using WB. These results may contribute to the reduction of the cost of enzymes used in the production of cellulosic ethanol or other biotechnological applications.

  18. EFSA CEF Panel (EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids), 2014. Scientific Opinion on xylanase from a genetically modified strain of Aspergillus oryzae (strain NZYM-FB)

    DEFF Research Database (Denmark)

    Poulsen, Morten; Binderup, Mona-Lise; Hallas-Møller, Torben

    . The xylanase is intended to be used in a number of food manufacturing processes, such as starch processing, beverage alcohol (distilling), brewing, baking and other cereal based processes. The dietary exposure was assessed according to the Budget method. The food enzyme did not induce gene mutations...

  19. A xylanase with broad pH and temperature adaptability from Streptomyces megasporus DSM 41476, and its potential application in brewing industry.

    Science.gov (United States)

    Qiu, Zhenhua; Shi, Pengjun; Luo, Huiying; Bai, Yingguo; Yuan, Tiezheng; Yang, Peilong; Liu, Suchun; Yao, Bin

    2010-05-05

    A xylanase gene, xynAM6, was isolated from the genomic DNA library of Streptomyces megasporus DSM 41476 using colony PCR screening method. The 1440-bp full-length gene encodes a 479-amino acid peptide consisting of a putative signal peptide of 36 residues, a family 10 glycoside hydrolase domain and a family 2 carbohydrate-binding module. The mature peptide of xynAM6 was successfully expressed in Pichia pastoris GS115. The optimal pH and temperature were pH 5.5 and 70°C, respectively. The enzyme showed broad temperature adaptability (>60% of the maximum activity at 50-80°C), had good thermostability at 60°C and 70°C, remained stable at pH 4.0-11.0, and was resistant to most proteases. The Km and Vmax values for oat spelt xylan were 1.68mgml(-1) and 436.76μmolmin(-1)mg(-1), respectively, and 2.33mgml(-1) and 406.93μmolmin(-1)mg(-1) for birchwood xylan, respectively. The hydrolysis products of XYNAM6 were mainly xylose and xylobiose. Addition of XYNAM6 (80U) to the brewery mash significantly reduced the filtration rate and viscosity by 36.33% and 35.51%, respectively. These favorable properties probably make XYNAM6 a good candidate for application in brewing industry. Copyright © 2010 Elsevier Inc. All rights reserved.

  20. Enzymatic hydrolysis of pretreated Alfa fibers (Stipa tenacissima) using β-d-glucosidase and xylanase of Talaromyces thermophilus from solid-state fermentation.

    Science.gov (United States)

    Mallek-Fakhfakh, Hanen; Fakhfakh, Jawhar; Walha, Kamel; Hassairi, Hajer; Gargouri, Ali; Belghith, Hafedh

    2017-10-01

    This work aims at realizing an optimal hydrolysis of pretreated Alfa fibers (Stipa tenacissima) through the use of enzymes produced from Talaromyces thermophilus AX4, namely β-d-glucosidase and xylanase, by a solid state fermentation process of an agro-industrial waste (wheat bran supplemented with lactose). The carbon source was firstly selected and the optimal values of three other parameters were determined: substrate loading (10g), moisture content (85%) and production time (10days); which led to an optimized enzymatic juice. The outcome was then supplemented with cellulases of T. reesei and used to optimize the enzymatic saccharification of alkali-pretreated Alfa fibers (PAF). The maximum saccharification yield of 83.23% was achieved under optimized conditions (substrate concentration 3.7% (w/v), time 144h and enzyme loading of 0.8 FPU, 15U CMCase, 60U β-d-glucosidase and 125U xylanase).The structural modification of PAF due to enzymatic saccharification was supported by the changes of morphologic and chemical composition observed through macroscopic representation, FTIR and X-Ray analysis. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Enhancing cellulase production by overexpression of xylanase regulator protein gene, xlnR, in Talaromyces cellulolyticus cellulase hyperproducing mutant strain.

    Science.gov (United States)

    Okuda, Naoyuki; Fujii, Tatsuya; Inoue, Hiroyuki; Ishikawa, Kazuhiko; Hoshino, Tamotsu

    2016-10-01

    We obtained strains with the xylanase regulator gene, xlnR, overexpressed (HXlnR) and disrupted (DXlnR) derived from Talaromyces cellulolyticus strain C-1, which is a cellulase hyperproducing mutant. Filter paper degrading enzyme activity and cellobiohydrolase I gene expression was the highest in HXlnR, followed by C-1 and DXlnR. These results indicate that the enhancement of cellulase productivity was succeeded by xlnR overexpression.

  2. Effects of xylanase and citric acid on the performance, nutrient retention, and characteristics of gastrointestinal tract of broilers fed low-phosphorus wheat-based diets

    NARCIS (Netherlands)

    Esmaeilipour, O.; Shivazad, M.; Moravej, H.; Aminzadeh, S.; Rezaian, M.; Krimpen, van M.M.

    2011-01-01

    An experiment was conducted to study the effects of xylanase and citric acid on the performance, nutrient retention, jejunal viscosity, and size and pH of the gastrointestinal tract of broilers fed a low-P wheat-based diet. The experiment was conducted as a 2 × 3 factorial arrangement with 2 levels

  3. Generation of transgenic wheat (Triticum aestivum L.) accumulating heterologous endo-xylanase or ferulic acid esterase in the endosperm.

    Science.gov (United States)

    Harholt, Jesper; Bach, Inga C; Lind-Bouquin, Solveig; Nunan, Kylie J; Madrid, Susan M; Brinch-Pedersen, Henrik; Holm, Preben B; Scheller, Henrik V

    2010-04-01

    Endo-xylanase (from Bacillus subtilis) or ferulic acid esterase (from Aspergillus niger) were expressed in wheat under the control of the endosperm-specific 1DX5 glutenin promoter. Constructs both with and without the endoplasmic reticulum retention signal (Lys-Asp-Glu-Leu) KDEL were used. Transgenic plants were recovered in all four cases but no qualitative differences could be observed whether KDEL was added or not. Endo-xylanase activity in transgenic grains was increased between two and threefold relative to wild type. The grains were shrivelled and had a 25%-33% decrease in mass. Extensive analysis of the cell walls showed a 10%-15% increase in arabinose to xylose ratio, a 50% increase in the proportion of water-extractable arabinoxylan, and a shift in the MW of the water-extractable arabinoxylan from being mainly larger than 85 kD to being between 2 and 85 kD. Ferulic acid esterase-expressing grains were also shrivelled, and the seed weight was decreased by 20%-50%. No ferulic acid esterase activity could be detected in wild-type grains whereas ferulic acid esterase activity was detected in transgenic lines. The grain cell walls had 15%-40% increase in water-unextractable arabinoxylan and a decrease in monomeric ferulic acid between 13% and 34%. In all the plants, the observed changes are consistent with a plant response that serves to minimize the effect of the heterologously expressed enzymes by increasing arabinoxylan biosynthesis and cross-linking.

  4. Generation of transgenic wheat (Triticum aestivum L.) accumulating heterologous endo-xylanase or ferulic acid esterase in the endosperm

    Energy Technology Data Exchange (ETDEWEB)

    Harholt, Jesper; Bach, Inga C; Lind-Bouquin, Solveig; Nunan, Kylie J.; Madrid, Susan M.; Brinch-Pedersen, Henrik; Holm, Preben B.; Scheller, Henrik V.

    2009-12-08

    Endo-xylanase (from Bacillus subtilis) or ferulic acid esterase (from Aspergillus niger) were expressed in wheat under the control of the endosperm specific 1DX5 glutenin promoter. Constructs both with and without the endoplasmic reticulum retention signal KDEL were used. Transgenic plants were recovered in all four cases but no qualitative differences could be observed whether KDEL was added or not. Endo-xylanase activity in transgenic grains was increased between two and three fold relative to wild type. The grains were shriveled and had a 25-33% decrease in mass. Extensive analysis of the cell walls showed a 10-15% increase in arabinose to xylose ratio, a 50% increase in the proportion of water extractable arabinoxylan, and a shift in the MW of the water extractable arabinoxylan from being mainly larger than 85 kD to being between 2 kD and 85 kD. Ferulic acid esterase expressing grains were also shriveled and the seed weight was decreased by 20-50%. No ferulic acid esterase activity could be detected in wild type grains whereas ferulic acid esterase activity was detected in transgenic lines. The grain cell walls had 15-40% increase in water unextractable arabinoxylan and a decrease in monomeric ferulic acid between 13 and 34%. In all the plants the observed changes are consistent with a plant response that serves to minimize the effect of the heterologously expressed enzymes by increasing arabinoxylan biosynthesis and cross-linking.

  5. Effect of Different Pretreatment of Sugar Cane Bagasse on Cellulase and Xylanases Production by the Mutant Penicillium echinulatum 9A02S1 Grown in Submerged Culture

    Directory of Open Access Journals (Sweden)

    Marli Camassola

    2014-01-01

    Full Text Available The main limitation to the industrial scale hydrolysis of cellulose is the cost of cellulase production. This study evaluated cellulase and xylanase enzyme production by the cellulolytic mutant Penicillium echinulatum 9A02S1 using pretreated sugar cane bagasse as a carbon source. Most cultures grown with pretreated bagasse showed similar enzymatic activities to or higher enzymatic activities than cultures grown with cellulose or untreated sugar cane bagasse. Higher filter paper activity (1.253 ± 0.147 U·mL−1 was detected in the medium on the sixth day of cultivation when bagasse samples were pretreated with sodium hydroxide, hydrogen peroxide, and anthraquinone. Endoglucanase enzyme production was also enhanced by pretreatment of the bagasse. Nine cultures grown with bagasse possessed higher β-glucosidase activities on the sixth day than the culture grown with cellulose. The highest xylanase activity was observed in cultures with cellulose and with untreated sugar cane bagasse. These results indicate that pretreated sugar cane bagasse may be able to serve as a partial or total replacement for cellulose in submerged fermentation for cellulase production using P. echinulatum, which could potentially reduce future production costs of enzymatic complexes capable of hydrolyzing lignocellulosic residues to form fermented syrups.

  6. Experimental mixture design as a tool to enhance glycosyl hydrolases production by a new Trichoderma harzianum P49P11 strain cultivated under controlled bioreactor submerged fermentation.

    Science.gov (United States)

    Delabona, Priscila da Silva; Farinas, Cristiane Sanchez; Lima, Deise Juliana da Silva; Pradella, José Geraldo da Cruz

    2013-03-01

    This work investigates the glycosyl hydrolase (GH) profile of a new Trichoderma harzianum strain cultivated under controlled bioreactor submerged fermentation. The influence of different medium components (delignified steam-exploded sugarcane bagasse, sucrose, and soybean flour) on GH biosynthesis was assessed using experimental mixture design (EMD). Additionally, the effect of increased component concentrations in culture media selected from the EMD was studied. It was found that that a mixed culture medium could significantly maximize GH biosynthesis rate, especially for xylanase enzymes which achieved a 2-fold increment. Overall, it was demonstrated that T. harzianumP49P11 enzymes have a great potential to be used in the deconstruction of biomass. Copyright © 2012 Elsevier Ltd. All rights reserved.

  7. THE INFLUENCE OF ASOCIATED SUPPLEMENT OF ALFA AMYLASE AND XYLANASE ON THE RHEOLOGY OF DOUGH CONCEARNING ITS CONSTITOGRAPHICAL PARAMETERS

    Directory of Open Access Journals (Sweden)

    RODICA CHEREJI

    2009-05-01

    Full Text Available In this paper we determined the influence of associated supplement of alfa amylase and xylanase on the rheology of dough concearning its constitographical parameters : maximum pressure (Pr max, (mb and the absorbed water (Wa, %. The analysis on the consistograph were conducted for constant hydration at the consistency of 500 UF. Determinations were made on 4 types of flour and optimal dosages were found for each enzyme, after which we prepared the optimal dosage of the enzymes in the compund for flour F1 and F2 : P1-840000 U. SKB/100 kg flour +8100 U. FXU /100 kg flour, P2-840000 U. SKB/100 kg flour+16200 U. FXU, /100 kg flour , P3-840000 U. SKB/100 kg flour+24300 U. FXU/100 kg flour , and for F3 and F4 thus: P1-280000 U. SKB/100 kg flour +8100 U. FXU /100 kg flour, P2- 280000 U. SKB/100 kg flour+16200 U. FXU/100 kg flour, P3-280000 U. SKB/100 kg flour+24300 U. FXU/100 kg flour. Fungous α-amylase and xylanase were used in these concentrations to establish which one is more apropriate to be added in flour to obtain superior quality of bread: finer texture of the crumb, prolongation of the freashness of the bread, improvind the colour and flavour, emproving the slicing ability.

  8. Xylanase Production from Trichoderma harzianum 1073 D3 with Alternative Carbon and Nitrogen Sources

    Directory of Open Access Journals (Sweden)

    Isil Seyis

    2005-01-01

    Full Text Available The effect of some natural wastes (orange pomace, orange peel, lemon pomace, lemon peel, apple pomace, pear peel, banana peel, melon peel and hazelnut shell on the production of xylanase from Trichoderma harzianum 1073 D3 has been studied and maximum activity has been observed on melon peel (26.5 U/mg of protein followed by apple pomace and hazelnut shell. Also, molasses could be used as an additional carbon source as it decreased the production time approximately by 50 %. Finally, potential alternatives of organic nitrogen source (cotton leaf and soybean residue wastes were analyzed and it was concluded that peptone could be replaced with these residues especially when economics of the process is the major objective.

  9. Effect of penicillium mutation by UV and gamma radiation on xylanase production

    International Nuclear Information System (INIS)

    Bakri, Y.; Shamma, M.; Hammoudeh, A.; Sharabi, N.

    2007-07-01

    Many microorganisms produce enzymes which have importance in industrial processes. Usually this production, is not sufficient for these needs at economical level. The bioindustry concentrates on increasing the production of these enzymes. This leads to the progress of this kind of industry, which use different biotechnology means, for example mutation and screening to choice more potent strain. In this study Ultra Violet and Gamma irradiation conducted on Penicillium canescen in order to produce new mutant strains, have the ability to produce more xylanase enzyme for industrial uses. Ultra Violet irradiation enable to select five mutant strains having more enzyme production ability. The best mutant strain PCUV12 (159 unit/ml) was 40% higher than the mother strain, at the dose 150.72 j/cm 2 . Gamma radiation produced new mutant strain PCGR6 which produced 26% more enzyme than the mother strain at dose 250 Gy.(author)

  10. Isolation, screening, and identification of potential cellulolytic and xylanolytic producers for biodegradation of untreated oil palm trunk and its application in saccharification of lemongrass leaves.

    Science.gov (United States)

    Ang, S K; Yahya, Adibah; Abd Aziz, Suraini; Md Salleh, Madihah

    2015-01-01

    This study presents the isolation and screening of fungi with excellent ability to degrade untreated oil palm trunk (OPT) in a solid-state fermentation system (SSF). Qualitative assay of cellulases and xylanase indicates notable secretion of both enzymes by 12 fungal strains from a laboratory collection and 5 strains isolated from a contaminated wooden board. High production of these enzymes was subsequently quantified in OPT in SSF. Aspergillus fumigates SK1 isolated from cow dung gives the highest xylanolytic activity (648.448 U g(-1)), generally high cellulolytic activities (CMCase: 48.006, FPase: 6.860, beta-glucosidase: 16.328 U g(-1)) and moderate lignin peroxidase activity (4.820 U/g), and highest xylanolytic activity. The xylanase encoding gene of Aspergillus fumigates SK1 was screened using polymerase chain reaction by a pair of degenerate primers. Through multiple alignment of the SK1 strain's xylanase nucleotide sequences with other published xylanases, it was confirmed that the gene belonged to the xylanase glycoside hydrolase family 11 (GH11) with a protein size of 24.49 kD. Saccharification of lemongrass leaves using crude cellulases and xylanase gives the maximum reducing sugars production of 6.84 g/L with glucose as the major end product and traces of phenylpropanic compounds (vanillic acid, p-coumaric acid, and ferulic acid).

  11. Use of Residual Biomass from the Textile Industry as Carbon Source for Production of a Low-Molecular-Weight Xylanase from Aspergillus oryzae

    Directory of Open Access Journals (Sweden)

    Gilvan Caetano Duarte

    2012-10-01

    Full Text Available Pretreated dirty cotton residue (PDCR from the textile industry was used as an alternative carbon source for the submerged cultivation of Aspergillus oryzae and the production of xylanases. The filtered culture supernatant was fractionated by ultrafiltration followed by three chromatographic steps, which resulted in the isolation of a homogeneous low-molecular-weight xylanase (Xyl-O1 with a mass of 21.5 kDa as determined by sodium dodecyl sulfate-polyacrilamide gel electrophoresis (SDS-PAGE co-polymerized with 0.1% oat spelt xylan. Enzyme catalysis was the most efficient at 50 °C and pH 6.0. The Km values (mg·mL−1 for the soluble fraction of oat spelt and birchwood xylans were 10.05 and 3.34, respectively. Xyl-O1 was more stable in the presence of 5,5-dithio-bis-(2-nitrobenzoic acid (DTNB, 1,4-dithiothreitol (DTT, l-cysteine or β-mercaptoethanol, which increased the rate of catalysis by 40%, 14%, 40% or 37%, respectively. The enzyme stability was improved at pH 7.0 in the presence of 20 mM l-cysteine, with the retention of nearly 100% of the activity after 6 h at 50 °C. Xyl-O1 catalyzed the cleavage of internal β-1,4 linkages of the soluble substrates containing d-xylose residues, with a maximum efficiency of 33% for the hydrolysis of birchwood xylan after 12 h of incubation. Identification of the hydrolysis products by high-performance anion exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD indicated the predominance of the hydrolysis products X2-X6 during the first 12 h of incubation and the accumulation of higher xylooligomers after the elution of the last xylooligomer standard, xylohexaose.

  12. Biological synthesis of Au nanoparticles using liquefied mash of cassava starch and their functionalization for enhanced hydrolysis of xylan by recombinant xylanase.

    Science.gov (United States)

    Zeng, Sumei; Du, Liangwei; Huang, Meiying; Feng, Jia-Xun

    2016-05-01

    Au nanoparticles (AuNPs) have shown the potential for a variety of applications due to their unique physical and chemical properties. In this study, a facile and affordable method for the synthesis of AuNPs via the liquefied mash of cassava starch has been described and the functionalized AuNPs by L-cysteine improved activity of recombinant xylanase was demonstrated. UV-Vis absorption spectroscopy, transmission electron microscopy, and zeta potential measurements were performed to characterize the AuNPs and monitor their synthesis. The presence of Au was confirmed by energy-dispersive X-ray spectroscopy (EDX) and the X-ray diffraction patterns showed that Au nanocrystals were face-centered cubic. The C=O stretching vibration in the Fourier transform infrared spectrum of AuNPs suggested that the hemiacetal C-OH of sugar molecules performed the reduction of Au³⁺ to Au⁰. The presence of C and O in the EDX spectrum and the negative zeta potential of AuNPs suggested that the biomolecules present in liquefied cassava mash were responsible for the stabilization of AuNPs. The surface of AuNPs was easily functionalized by L-cysteine, which improved the stability of AuNPs. Moreover, cysteine-functionalized AuNPs could significantly improve recombinant xylanase efficiency and stability.

  13. Thermal and chemical denaturation of Bacillus circulans xylanase: A biophysical chemistry laboratory module.

    Science.gov (United States)

    Raabe, Richard; Gentile, Lisa

    2008-11-01

    A number of institutions have been, or are in the process of, modifying their biochemistry major to include some emphasis on the quantitative physical chemistry of biomolecules. Sometimes this is done as a replacement for part for the entire physical chemistry requirement, while at other institutions this is incorporated as a component into the traditional two-semester biochemistry series. The latter is the model used for biochemistry and molecular biology majors at the University of Richmond, whose second semester of biochemistry is a course entitled Proteins: Structure, Function, and Biophysics. What is described herein is a protein thermodynamics laboratory module, using the protein Bacillus circulans xylanase, which reinforces many lecture concepts, including: (i) the denatured (D) state ensemble of a protein can be different, depending on how it was populated; (ii) intermediate states may be detected by some spectroscopic techniques but not by others; (iii) the use and assumptions of the van't Hoff approach to calculate ΔH(o) , ΔS(o) , and ΔG(o) (T) for thermal protein unfolding transitions; and (iv) the use and assumptions of an approach that allows determination of the Gibb's free energy of a protein unfolding transition based on the linear dependence of ΔG(o) on the concentration of denaturant used. This module also requires students to design their own experimental protocols and spend time in the primary literature, both important parts of an upper division lab. Copyright © 2008 International Union of Biochemistry and Molecular Biology, Inc.

  14. Production of xylooligosaccharides from forest waste by membrane separation and Paenibacillus xylanase hydrolysis

    Directory of Open Access Journals (Sweden)

    Chun-Han Ko

    2013-02-01

    Full Text Available Xylooligosaccharides (XO, derived from the alkaline (NaOH extractant of Mikania micrantha, were produced using multiple staged membrane separation and enzymatic xylanolysis. Staged nanofiltration (NMX, ultrafiltration (EUMX, and centrifugation (EMX processes for the ethanol precipitates were conducted. NMX recovered 97.26% of total xylose and removed 73.18% of sodium ions. Concentrations of total xylose were raised from 10.98 to 51.85 mg/mL by the NMX process. Recovered xylan-containing solids were hydrolyzed by the recombinant Paenibacillus xylanase. 68% XO conversions from total xylose of NMX was achieved in 24 hours. Xylopentaose (DP 5 was the major product from NMX and EMX hydrolysis. Xylohexaose (DP 6 was the major product from EUMX hydrolysis. Results of the present study suggest the applicability for XO production by nanofiltration, as NMX gave higher XO yields compared to those from a conventional ethanol-related lignocellulosic waste conversion process.

  15. Potential application of waste from castor bean (Ricinus communis L.) for production for xylanase of interest in the industry.

    Science.gov (United States)

    Herculano, Polyanna Nunes; Moreira, Keila Aparecida; Bezerra, Raquel Pedrosa; Porto, Tatiana Souza; de Souza-Motta, Cristina Maria; Porto, Ana Lúcia Figueiredo

    2016-12-01

    Xylanases activity (XY) from Aspergillus japonicus URM5620 produced by Solid-State Fermentation (SSF) of castor press cake (Ricinus communis) on different conditions of production and extraction by PEG/citrate aqueous two-phase system (ATPS) were investigated. XY production was influenced by substrate amount (5-10 g), initial moisture (15-35 %), pH (4.0-6.0) and temperature (25-35 °C), obtaining the maximum activity of 29,085 ± 1808 U g ds -1 using 5.0 g of substrate with initial moisture of 15 % at 25 °C and pH 6.0, after 120 h of fermentation. The influence of PEG molar mass (1000-8000 g mol -1 ), phase concentrations (PEG 20.0-24.0 % w/w and sodium citrate 15-20 % w/w) and pH (6.0-8.0) on partition coefficient, purification factor, yield and selectivity of XY were determinate. Enzyme partitioning into the PEG rich phase was favored by M PEG 8000 (g mol -1 ), C PEG 24 % (w/w), C C 20 % (w/w) and pH 8.0, resulting in partition coefficient of 50.78, activity yield of 268 %, 7.20-fold purification factor and selectivity of 293. A. japonicus URM5620 has a potential role in the development of a bioprocess for the XY production using low-cost media. In addition, the present study proved it is feasible to extract xylanase from SSF by adopting the one step ATPS consisting of PEG/citrate.

  16. Response surface methodology optimization of partitioning of xylanase form Aspergillus Niger by metal affinity polymer-salt aqueous two-phase systems.

    Science.gov (United States)

    Fakhari, Mohamad Ali; Rahimpour, Farshad; Taran, Mojtaba

    2017-09-15

    Aqueous two phase affinity partitioning system using metal ligands was applied for partitioning and purification of xylanase produced by Aspergillus Niger. To minimization the number of experiments for the design parameters and develop predictive models for optimization of the purification process, response surface methodology (RSM) with a face-centered central composite design (CCF) has been used. Polyethylene glycol (PEG) 6000 was activated using epichlorohydrin, covalently linked to iminodiacetic acid (IDA), and the specific metal ligand Cu was attached to the polyethylene glycol-iminodiacetic acid (PEG-IDA). The influence of some experimental variables such as PEG (10-18%w/w), sodium sulfate (8-12%), PEG-IDA-Cu 2+ concentration (0-50% w/w of total PEG), pH of system (4-8) and crude enzyme loading (6-18%w/w) on xylanase and total protein partitioning coefficient, enzyme yield and enzyme specific activity were systematically evaluated. Two optimal point with high enzyme partitioning factor 10.97 and yield 79.95 (including 10% PEG, 12% Na 2 SO 4 , 50% ligand, pH 8 and 6% crude enzyme loading) and high specific activity in top phase 42.21 (including 14.73% PEG, 8.02% Na 2 SO 4 , 28.43% ligand, pH 7.7 and 6.08% crude enzyme loading) were attained. The adequacy of the RSM models was verified by a good agreement between experimental and predicted results. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. On the sensitivity of protein data bank normal mode analysis: an application to GH10 xylanases

    Science.gov (United States)

    Tirion, Monique M.

    2015-12-01

    Protein data bank entries obtain distinct, reproducible flexibility characteristics determined by normal mode analyses of their three dimensional coordinate files. We study the effectiveness and sensitivity of this technique by analyzing the results on one class of glycosidases: family 10 xylanases. A conserved tryptophan that appears to affect access to the active site can be in one of two conformations according to x-ray crystallographic electron density data. The two alternate orientations of this active site tryptophan lead to distinct flexibility spectra, with one orientation thwarting the oscillations seen in the other. The particular orientation of this sidechain furthermore affects the appearance of the motility of a distant, C terminal region we term the mallet. The mallet region is known to separate members of this family of enzymes into two classes.

  18. On the sensitivity of protein data bank normal mode analysis: an application to GH10 xylanases

    International Nuclear Information System (INIS)

    Tirion, Monique M

    2015-01-01

    Protein data bank entries obtain distinct, reproducible flexibility characteristics determined by normal mode analyses of their three dimensional coordinate files. We study the effectiveness and sensitivity of this technique by analyzing the results on one class of glycosidases: family 10 xylanases. A conserved tryptophan that appears to affect access to the active site can be in one of two conformations according to x-ray crystallographic electron density data. The two alternate orientations of this active site tryptophan lead to distinct flexibility spectra, with one orientation thwarting the oscillations seen in the other. The particular orientation of this sidechain furthermore affects the appearance of the motility of a distant, C terminal region we term the mallet. The mallet region is known to separate members of this family of enzymes into two classes. (paper)

  19. Molecular Modeling and MM-PBSA Free Energy Analysis of Endo-1,4-β-Xylanase from Ruminococcus albus 8

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    Dongling Zhan

    2014-09-01

    Full Text Available Endo-1,4-β-xylanase (EC 3.2.1.8 is the enzyme from Ruminococcus albus 8 (R. albus 8 (Xyn10A, and catalyzes the degradation of arabinoxylan, which is a major cell wall non-starch polysaccharide of cereals. The crystallographic structure of Xyn10A is still unknown. For this reason, we report a computer-assisted homology study conducted to build its three-dimensional structure based on the known sequence of amino acids of this enzyme. In this study, the best similarity was found with the Clostridium thermocellum (C. thermocellum N-terminal endo-1,4-β-d-xylanase 10 b. Following the 100 ns molecular dynamics (MD simulation, a reliable model was obtained for further studies. Molecular Mechanics/Poisson-Boltzmann Surface Area (MM-PBSA methods were used for the substrate xylotetraose having the reactive sugar, which was bound in the −1 subsite of Xyn10A in the 4C1 (chair and 2SO (skew boat ground state conformations. According to the simulations and free energy analysis, Xyn10A binds the substrate with the −1 sugar in the 2SO conformation 39.27 kcal·mol−1 tighter than the substrate with the sugar in the 4C1 conformation. According to the Xyn10A-2SO Xylotetraose (X4(sb interaction energies, the most important subsite for the substrate binding is subsite −1. The results of this study indicate that the substrate is bound in a skew boat conformation with Xyn10A and the −1 sugar subsite proceeds from the 4C1 conformation through 2SO to the transition state. MM-PBSA free energy analysis indicates that Asn187 and Trp344 in subsite −1 may an important residue for substrate binding. Our findings provide fundamental knowledge that may contribute to further enhancement of enzyme performance through molecular engineering.

  20. Production of Cellulases, Xylanase, Pectinase, alpha-amylase and Protease Enzymes Cocktail by Bacillus spp. and Their Mixed Cultures with Candida tropicalis and Rhodotorula glutinis under Solid State Fermentation

    International Nuclear Information System (INIS)

    El-Batal, A.I.; Abo-State, M.A.

    2006-01-01

    A group of twelve locally isolated Bacillus species, B.megaterium (MAI and MA II), B.licheniformis (MLI and ML II); B. circulans, B. stearothermophilis, B.cereus, B.sphaericus, B. pumilus, B. laterosporus, B. coagulans and B. pantothenticus, were examined for the production of cellulases, xylanase, pectinase, alpha-amylase and protease enzymes cocktail on wheat bran under solid state fermentation (SSF). All species were found to be potent hydrolyzing enzymes producers and the superior producing species were B. megaterium MAI and B. licheniformis. On the other hand, both of them still produced highest enzyme titres when mixed with Candida tropicalis or Rhodotorula glutinis, yeast strains. The two superior bacterial strains produced the highest enzymatic activities when coculturing with C. tropicalis compared with coculturing with R. glutinis only or with both C. tropicalis and R. glutinis in combination. The inferior activities of cocultures (B. megaterinm MAI and R. glutinis) were enhanced in carboxymethyl cellulase (CMCase), filter paper cellulase (FPase), avecilase, xylanase, pectinase, -amylase and protease by gamma irradiation at dose 1.0 kGy with percent increase 8 %, 20 %, 10 %, 4 %, 31 %, 22 % and 34 %, respectively as compared with un-irradiated cocultures

  1. Effect of cellulase, xylanase and α-amylase combinations on the rheological properties of Chinese steamed bread dough enriched in wheat bran.

    Science.gov (United States)

    Liu, Wenjun; Brennan, Margaret Anne; Serventi, Luca; Brennan, Charles Stephen

    2017-11-01

    The present study investigates the effects of α-amylase (6 and 10ppm), xylanase (70 and 120ppm) and cellulase (35 and 60ppm) on the rheological properties of bread dough. The mixing property of dough was measured by using a DoughLAB. The extension and stickiness of dough were analysed using the Texture Analyzer. The results illustrate that the addition of single enzyme and enzyme combinations can increase the extensibility, softening, mixing tolerance index (MTI) and stickiness, whereas decrease the resistance to extension. For water absorption, the addition of single enzyme had no significant effect, while the combination enzyme significantly (pcellulase had a synergetic effect on the dough rheology. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Generation of transgenic wheat (Triticum aestivum L.) accumulating heterologous endo-xylanase or ferulic acid esterase in the endosperm

    DEFF Research Database (Denmark)

    Harholt, Jesper; Bach, Inga Christensen; Lind Bouquin, Solveig

    2010-01-01

    Endo-xylanase (from Bacillus subtilis) or ferulic acid esterase (from Aspergillus niger) were expressed in wheat under the control of the endosperm-specific 1DX5 glutenin promoter. Constructs both with and without the endoplasmic reticulum retention signal (Lys-Asp-Glu-Leu) KDEL were used....... Extensive analysis of the cell walls showed a 10%-15% increase in arabinose to xylose ratio, a 50% increase in the proportion of water-extractable arabinoxylan, and a shift in the MW of the water-extractable arabinoxylan from being mainly larger than 85 kD to being between 2 and 85 kD. Ferulic acid esterase......-expressing grains were also shrivelled, and the seed weight was decreased by 20%-50%. No ferulic acid esterase activity could be detected in wild-type grains whereas ferulic acid esterase activity was detected in transgenic lines. The grain cell walls had 15%-40% increase in water-unextractable arabinoxylan...

  3. ORF Sequence: NC_006177 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available Symbiobacterium thermophilum IAM 14863] MTLVRDAMIQVEPLSPNDPIFVALQAMRKQRLPFVPVVRSDGTLYGLVTEGDLVRLIYRATRESGESVPHWIRGAGRQVLLVQSVKEVVTRELDTIGPDIPLEDAAELMVRNHRKVLPVVADGRPMGYLMRSAVVDHLIGE

  4. A family 30 glucurono-xylanase from Bacillus subtilis LC9: Expression, characterization and its application in Chinese bread making.

    Science.gov (United States)

    Guo, Yalan; Gao, Zhen; Xu, Jiaxing; Chang, Siyuan; Wu, Bin; He, Bingfang

    2018-05-22

    A GH30-8 endoxylanase was identified from an environmental Bacillus subtilis isolate following growth selection on aspen wood glucuronoxylan. The putative endoxylanase was cloned for protein expression and characterization in the Gram-positive protease deficient protein expression host B. subtilis WB800. The extracellular activity obtained was 55 U/mL, which was 14.5-fold higher than that obtained with the native species. The apparent molecular mass of BsXyn30 was estimated as 43 kDa by SDS-PAGE. BsXyn30 showed an optimal activity at pH 7.0 and 60 °C. Recombinant BsXyn30 displayed maximum activity against aspen wood xylan, followed by beechwood xylan but showed no catalytic activity on arabinose-substituted xylans. Analysis of hydrolyzed products of beechwood xylan by thin-layer chromatography and mass spectroscopy revealed the presence of xylooligosaccharides with a single methyl-glucuronic acid residue. BsXyn30 exhibited very low activity for hydrolysis xylotetraose and xylopentaose, but had no detectable activity against xylobiose and xylotriose. Using BsXyn30 as an additive in breadmaking, a decrease in water-holding capacity, an increase in dough expansion as well as improvements in volume and specific volume of the bread were recorded. Thus, the present study provided the basis for the application of GH30 xylanase in breadmaking. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. Characterization of lignocellulolytic activities from fungi isolated from the deep-sea sponge Stelletta normani.

    Directory of Open Access Journals (Sweden)

    Ramón Alberto Batista-García

    Full Text Available Extreme habitats have usually been regarded as a source of microorganisms that possess robust proteins that help enable them to survive in such harsh conditions. The deep sea can be considered an extreme habitat due to low temperatures (<5°C and high pressure, however marine sponges survive in these habitats. While bacteria derived from deep-sea marine sponges have been studied, much less information is available on fungal biodiversity associated with these sponges. Following screening of fourteen fungi isolated from the deep-sea sponge Stelletta normani sampled at a depth of 751 metres, three halotolerant strains (TS2, TS11 and TS12 were identified which displayed high CMCase and xylanase activities. Molecular based taxonomic approaches identified these strains as Cadophora sp. TS2, Emericellopsis sp. TS11 and Pseudogymnoascus sp. TS 12. These three fungi displayed psychrotolerance and halotolerant growth on CMC and xylan as sole carbon sources, with optimal growth rates at 20°C. They produced CMCase and xylanase activities, which displayed optimal temperature and pH values of between 50-70°C and pH 5-8 respectively, together with good thermostability and halotolerance. In solid-state fermentations TS2, TS11 and TS12 produced CMCases, xylanases and peroxidase/phenol oxidases when grown on corn stover and wheat straw. This is the first time that CMCase, xylanase and peroxidase/phenol oxidase activities have been reported in these three fungal genera isolated from a marine sponge. Given the biochemical characteristics of these ligninolytic enzymes it is likely that they may prove useful in future biomass conversion strategies involving lignocellulosic materials.

  6. Characterization of lignocellulolytic activities from fungi isolated from the deep-sea sponge Stelletta normani.

    Science.gov (United States)

    Batista-García, Ramón Alberto; Sutton, Thomas; Jackson, Stephen A; Tovar-Herrera, Omar Eduardo; Balcázar-López, Edgar; Sánchez-Carbente, María Del Rayo; Sánchez-Reyes, Ayixon; Dobson, Alan D W; Folch-Mallol, Jorge Luis

    2017-01-01

    Extreme habitats have usually been regarded as a source of microorganisms that possess robust proteins that help enable them to survive in such harsh conditions. The deep sea can be considered an extreme habitat due to low temperatures (<5°C) and high pressure, however marine sponges survive in these habitats. While bacteria derived from deep-sea marine sponges have been studied, much less information is available on fungal biodiversity associated with these sponges. Following screening of fourteen fungi isolated from the deep-sea sponge Stelletta normani sampled at a depth of 751 metres, three halotolerant strains (TS2, TS11 and TS12) were identified which displayed high CMCase and xylanase activities. Molecular based taxonomic approaches identified these strains as Cadophora sp. TS2, Emericellopsis sp. TS11 and Pseudogymnoascus sp. TS 12. These three fungi displayed psychrotolerance and halotolerant growth on CMC and xylan as sole carbon sources, with optimal growth rates at 20°C. They produced CMCase and xylanase activities, which displayed optimal temperature and pH values of between 50-70°C and pH 5-8 respectively, together with good thermostability and halotolerance. In solid-state fermentations TS2, TS11 and TS12 produced CMCases, xylanases and peroxidase/phenol oxidases when grown on corn stover and wheat straw. This is the first time that CMCase, xylanase and peroxidase/phenol oxidase activities have been reported in these three fungal genera isolated from a marine sponge. Given the biochemical characteristics of these ligninolytic enzymes it is likely that they may prove useful in future biomass conversion strategies involving lignocellulosic materials.

  7. Mathematical modeling of enzyme production using Trichoderma harzianum P49P11 and sugarcane bagasse as carbon source.

    Science.gov (United States)

    Gelain, Lucas; da Cruz Pradella, José Geraldo; da Costa, Aline Carvalho

    2015-12-01

    A mathematical model to describe the kinetics of enzyme production by the filamentous fungus Trichoderma harzianum P49P11 was developed using a low cost substrate as main carbon source (pretreated sugarcane bagasse). The model describes the cell growth, variation of substrate concentration and production of three kinds of enzymes (cellulases, beta-glucosidase and xylanase) in different sugarcane bagasse concentrations (5; 10; 20; 30; 40 gL(-1)). The 10 gL(-1) concentration was used to validate the model and the other to parameter estimation. The model for enzyme production has terms implicitly representing induction and repression. Substrate variation was represented by a simple degradation rate. The models seem to represent well the kinetics with a good fit for the majority of the assays. Validation results indicate that the models are adequate to represent the kinetics for a biotechnological process. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Properties of Enzyme Pretreated Wikstroemia sikokiana and Broussonetia papyrifera Bast Fiber Pulps

    Directory of Open Access Journals (Sweden)

    Lang-Dong Lin

    2015-04-01

    Full Text Available Xylanase, pectinase complex, and BL11 pectinase were employed for the pretreatment of gampi and paper mulberry bast fiber pulps prior to chlorine dioxide bleaching. The bleaching efficiencies of the pulps with different enzymatic pretreatments were investigated. Accelerated aging by heat-humidity treatment was also conducted to evaluate yellowing phenomena and to estimate the prevention of brightness reversion (brightness retention by enzymatic pretreatment. The order of active chlorine required with respect to pretreatment was pectinase complex > xylanase > BL11 pectinase for soda and soda/oxalate gampi pulps and pectinase complex > BL11 pectinase > xylanase for soda and soda/oxalate paper mulberry pulps. Higher brightness retention values were observed for soda/oxalate pulps compared to soda pulps. The brightness retention levels for gampi pulps and mulberry pulps after ClO2 bleaching with enzymatic pretreatment were higher than the levels of ClO2 and NaClO bleaching pulps. Enzymatic treatments were thus able to reduce the usage of ClO2 and to assist in producing photo-stable paper materials for art and artifact-repairing applications. Thus, enzymatic pretreatment of the pulp has the potential to meet world trends and environmental sustainability for pulp and paper industries.

  9. Impact of orientation of carbohydrate binding modules family 22 and 6 on the catalytic activity of Thermotoga maritima xylanase XynB.

    Science.gov (United States)

    Tajwar, Razia; Shahid, Saher; Zafar, Rehan; Akhtar, Muhammad Waheed

    2017-11-01

    Xylanase XynB of the hyperthermophile Thermotoga maritima, which belongs to glycoside hydrolase family 10 (GH10), does not have an associated carbohydrate binding module (CBM) in the native state. CBM6 and CBM22 from a thermophile Clostridium thermocellum were fused to the catalytic domain of XynB (XynB-C) to determine the effects on activity and other properties. XynB-B22C and XynB-CB22, produced by fusing CBM22 to the N- and C-terminal of XynB-C, showed 1.7- and 3.24-fold increase in activity against the insoluble birchwood xylan, respectively. Similarly, CBM6 when attached to the C-terminal of XynB-C resulted in 2.0-fold increase in activity, whereas its attachment to the N-terminal did not show any increase of activity. XynB-B22C and XynB-CB22 retained all the activity, whereas XynB-B6C and XynB-CB6 lost 17 and 11% of activity, respectively, at 60°C for 4h. Thermostability data and the secondary structure contents obtained by molecular modelling are in agreement with the data from circular dichroism analysis. Molecular modelling analysis showed that the active site residues of the catalytic domain and the binding residues of CBM6 and CBM22 were located on the surface of molecule, except XynB-B6C, where the binding residues were found somewhat buried. In the case of XynB-CB22, the catalytic and the binding residues seem to be located favorably adjacent to each other, thus showing higher increase in activity. This study shows that the active site residues of the catalytic domain and the binding residues of the CBM are arranged in a unique fashion, not reported before. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. ORF Alignment: NC_006177 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... thermophilum IAM 14863] ... Length = 116 ... Query: 2 ... NQYKAYQVQTTAPEDQVALLYEGAQRFIDKAALALQQGRYG...EVSAYVGRAQQIFTELRTA 61 ... NQYKAYQVQTTAPEDQVALLYEGAQRFIDKAALALQQGRYGEVSAYV...GRAQQIFTELRTA Sbjct: 1 ... NQYKAYQVQTTAPEDQVALLYEGAQRFIDKAALALQQGRYGEVSAYVGRAQQIFTELRTA 60 ...

  11. ORF Alignment: NC_006177 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... [Symbiobacterium thermophilum IAM 14863] ... Length = 63 ... Query: 2 ... CSFCVDKIEHVDYKDAARLKRYITERGKILPRRISGNCARHQRQLTVAIKRARIMA...LLPF 61 ... CSFCVDKIEHVDYKDAARLKRYITERGKILPRRISGNCARHQRQLTVAIKRARIMA...LLPF Sbjct: 1 ... CSFCVDKIEHVDYKDAARLKRYITERGKILPRRISGNCARHQRQLTVAIKRARIMALLPF 60 ...

  12. ORF Alignment: NC_006177 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available in ... [Symbiobacterium thermophilum IAM 14863] ... Length = 138 ... Query: 4 ... VRDAMIQVEPLSPNDPIFVALQAMRKQRLPFVPV...VRSDGTLYGLVTEGDLVRLIYRATRE 63 ... VRDAMIQVEPLSPNDPIFVALQAMRKQRLPFVPV...VRSDGTLYGLVTEGDLVRLIYRATRE Sbjct: 1 ... VRDAMIQVEPLSPNDPIFVALQAMRKQRLPFVPVVRSDGTLYGLVTEGDLVRLIYRATR

  13. ORF Alignment: NC_006177 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... [Symbiobacterium thermophilum IAM 14863] ... Length = 217 ... Query: 15 ... AEVYRRLLEAFGPRHWWPSALEPPPAKAQPFEM...IAGAILVQNVAWSNAAKAVRALAEAGX 74 ... AEVYRRLLEAFGPRHWWPSALEPPPAKAQPFEM...IAGAILVQNVAWSNAAKAVRALAEAG ... Sbjct: 1 ... AEVYRRLLEAFGPRHWWPSALEPPPAKAQPFEMIAGAILVQNVAWSNAAKAVRALAEAGL 60 ... Q

  14. ORF Alignment: NC_006177 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available beta ... subunit [Symbiobacterium thermophilum IAM 14863] ... Length = 248 ... Query: 3 ... RAATWETWRPDPANTRQHVAMLVDTSECI...GCKGCEVACKQWNQNEARINQDPGTYQSHPA 62 ... RAATWETWRPDPANTRQHVAMLVDTSECI...GCKGCEVACKQWNQNEARINQDPGTYQSHPA Sbjct: 1 ... RAATWETWRPDPANTRQHVAMLVDTSECIGCKGCEVACKQWNQNEARI

  15. Author Details - African Journals Online

    African Journals Online (AJOL)

    Effects of salt and water stress on plant biomass and photosynthetic characteristics of Tamarisk (Tamarix chinensis Lour.) seedlings. Abstract PDF · Vol 11, No 15 (2012) - Articles Production and some properties of the thermostable feruloyl esterase and xylanase from Bacillus pumilus. Abstract PDF · Vol 11, No 26 (2012) - ...

  16. Expression of Critical Sulfur- and Iron-Oxidation Genes and the Community Dynamics During Bioleaching of Chalcopyrite Concentrate by Moderate Thermophiles.

    Science.gov (United States)

    Zhou, Dan; Peng, Tangjian; Zhou, Hongbo; Liu, Xueduan; Gu, Guohua; Chen, Miao; Qiu, Guanzhou; Zeng, Weimin

    2015-07-01

    Sulfate adenylyltransferase gene and 4Fe-4S ferredoxin gene are the key genes related to sulfur and iron oxidations during bioleaching system, respectively. In order to better understand the bioleaching and microorganism synergistic mechanism in chalcopyrite bioleaching by mixed culture of moderate thermophiles, expressions of the two energy metabolism genes and community dynamics of free and attached microorganisms were investigated. Specific primers were designed for real-time quantitative PCR to study the expression of these genes. Real-time PCR results showed that sulfate adenylyltransferase gene was more highly expressed in Sulfobacillus thermosulfidooxidans than that in Acidithiobacillus caldus, and expression of 4Fe-4S ferredoxin gene was higher in Ferroplasma thermophilum than that in S. thermosulfidooxidans and Leptospirillum ferriphilum. The results indicated that in the bioleaching system of chalcopyrite concentrate, sulfur and iron oxidations were mainly performed by S. thermosulfidooxidans and F. thermophilum, respectively. The community dynamics results revealed that S. thermosulfidooxidans took up the largest proportion during the whole period, followed by F. thermophilum, A. caldus, and L. ferriphilum. The CCA analysis showed that 4Fe-4S ferredoxin gene expression was mainly affected (positively correlated) by high pH and elevated concentration of ferrous ion, while no factor was observed to prominently influence the expression of sulfate adenylyltransferase gene.

  17. D-Xylose fermentation, xylitol production and xylanase activities by seven new species of Sugiyamaella.

    Science.gov (United States)

    Sena, Letícia M F; Morais, Camila G; Lopes, Mariana R; Santos, Renata O; Uetanabaro, Ana P T; Morais, Paula B; Vital, Marcos J S; de Morais, Marcos A; Lachance, Marc-André; Rosa, Carlos A

    2017-01-01

    Sixteen yeast isolates identified as belonging to the genus Sugiyamaella were studied in relation to D-xylose fermentation, xylitol production, and xylanase activities. The yeasts were recovered from rotting wood and sugarcane bagasse samples in different Brazilian regions. Sequence analyses of the internal transcribed spacer (ITS) region and the D1/D2 domains of large subunit rRNA gene showed that these isolates belong to seven new species. The species are described here as Sugiyamaella ayubii f.a., sp. nov. (UFMG-CM-Y607 T  = CBS 14108 T ), Sugiyamaella bahiana f.a., sp. nov. (UFMG-CM-Y304 T  = CBS 13474 T ), Sugiyamaella bonitensis f.a., sp. nov. (UFMG-CM-Y608 T  = CBS 14270 T ), Sugiyamaella carassensis f.a., sp. nov. (UFMG-CM-Y606 T  = CBS 14107 T ), Sugiyamaella ligni f.a., sp. nov. (UFMG-CM-Y295 T  = CBS 13482 T ), Sugiyamaella valenteae f.a., sp. nov. (UFMG-CM-Y609 T  = CBS 14109 T ) and Sugiyamaella xylolytica f.a., sp. nov. (UFMG-CM-Y348 T  = CBS 13493 T ). Strains of the described species S. boreocaroliniensis, S. lignohabitans, S. novakii and S. xylanicola, isolated from rotting wood of Brazilian ecosystems, were also compared for traits relevant to xylose metabolism. S. valenteae sp. nov., S. xylolytica sp. nov., S. bahiana sp. nov., S. bonitensis sp. nov., S. boreocarolinensis, S. lignohabitans and S. xylanicola were able to ferment D-xylose to ethanol. Xylitol production was observed for all Sugiyamaella species studied, except for S. ayubii sp. nov. All species studied showed xylanolytic activity, with S. xylanicola, S. lignohabitans and S. valenteae sp. nov. having the highest values. Our results suggest these Sugiyamaella species have good potential for biotechnological applications.

  18. Effect of exogenous xylanase, amylase, and protease as single or combined activities on nutrient digestibility and growth performance of broilers fed corn/soy diets.

    Science.gov (United States)

    Amerah, A M; Romero, L F; Awati, A; Ravindran, V

    2017-04-01

    Two trials (a 42-d performance and a 21-d cohort digestibility) were conducted to evaluate the performance and nutrient digestibility of broilers fed corn diets supplemented with exogenous xylanase, amylase, and protease as single or combined activities. A nutritionally adequate, positive control (PC) diet was formulated. The negative control (NC) diet was formulated to be lower in metabolizable energy (∼86 kcal/kg diet) and digestible amino acids (1 to 2%) compared to PC. The other 4 treatments were based on the NC and they were either supplemented with xylanase (X), amylase (A), protease (P), or a combination of X, A, and P (XAP; to provide 2,000 U of X, 200 U of A, and 4,000 U of P/kg diet). All diets were marginal in AvP and Ca and contained a background of phytase (1,000 FTU/kg). In each trial, male broiler (Ross 308) chicks were allocated to the 5 treatments (10 replicates of 20 birds/pen and 9 replicates of 8 birds/cage for the performance and digestibility trials, respectively). In the digestibility trial, ileal digesta was collected on d21 for the determination of nutrient utilization. Data were subjected to one-way ANOVA and means were separated by Tukey's HSD test. Only the XAP improved (P digestibility and apparent ileal digestible energy (AIDE). Both P and XAP improved N retention. The relative improvement in energy digestibility due to enzyme supplementation was greater at the ileal level than that measured in the excreta. The measured changes on AIDE due to supplemental enzymes were much higher than the sum of calculated contributions from starch, fat, and protein. Supplementation of all enzymes reduced (P  0.05) by dietary treatments. Both X and XAP had lower (P  0.05) FCR compared to PC. In conclusion, these results suggest a synergistic effect between X, A and P on broiler performance and nutrient digestibility. In the current study, AIDE measurements appeared to overestimate the enzyme response. Calculation of the energy contribution by

  19. ORF Alignment: NC_006177 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... [Symbiobacterium thermophilum IAM 14863] ... Length = 126 ... Query: 18 ... VVDAEGKTLGRLASKVAAILRGKHKPTFTPHVDC...GDFVIIVNAEKIQVTGKKVTDKVYYR 77 ... VVDAEGKTLGRLASKVAAILRGKHKPTFTPHVDC...GDFVIIVNAEKIQVTGKKVTDKVYYR Sbjct: 1 ... VVDAEGKTLGRLASKVAAILRGKHKPTFTPHVDCGDFVIIVNAEKIQVTGKKVTDKVYYR 60 ... Query: 138 PEPLEV 143 ... PEPLEV Sbjct: 121 PEPLEV 126

  20. Corn and soybean meal metabolizable energy with the addition of exogenous enzymes for poultry

    Directory of Open Access Journals (Sweden)

    LRB Dourado

    2009-03-01

    Full Text Available Two metabolism assays were carried out to determine corn and soybean meal metabolizable energy when enzymes were added. In the first trial, 35 cockerels per studied feedstuff (corn and soybean meal were distributed in a completely randomized experimental design with four treatments of seven replicates of one bird each. The evaluated treatments were: ingredient (corn and soybean meal with no enzyme addition, with the addition of an enzyme complex (xylanase, amylase, protease - XAP, xylanase, or phytase. Precise feeding method was used to determine true metabolizable energy corrected for nitrogen balance (TMEn. The use of enzymes did not result in any differences (p>0.05 in soybean meal TMEn, but phytase improved corn TMEn in 2.3% (p=0.004. In the second trial, 280 seven-day-old broiler chicks were distributed in a completely randomized experimental design with seven treatments of five replicates of eight birds each. Treatments consisted of corn with no enzyme addition or with the addition of amylase, xylanase, phytase, XAP complex, XAP+phytase combination, or xylanase/ pectinase/β-glucanase complex (XPBG. Corn was supplemented with macro and trace minerals. Total excreta collection was used to determine apparent metabolizable energy corrected for nitrogen balance (AMEn. Differences were observed (p=0.08 in AMEn and dry matter metabolizability coefficient (p=0.03. The combination of the XAP complex with phytase promoted a 2.11% increase in corn AMEn values, and the remaining enzymes allowed increased between 0.86% and 1.66%.

  1. Extraction of Glucuronoarabinoxylan from Quinoa Stalks (Chenopodium quinoa Willd.) and Evaluation of Xylooligosaccharides Produced by GH10 and GH11 Xylanases.

    Science.gov (United States)

    Salas-Veizaga, Daniel Martin; Villagomez, Rodrigo; Linares-Pastén, Javier A; Carrasco, Cristhian; Álvarez, María Teresa; Adlercreutz, Patrick; Nordberg Karlsson, Eva

    2017-10-04

    Byproducts from quinoa are not yet well explored sources of hemicellulose or products thereof. In this work, xylan from milled quinoa stalks was retrieved to 66% recovery by akaline extraction using 0.5 M NaOH at 80 °C, followed by ethanol precipitation. The isolated polymer eluted as a single peak in size-exclusion chromatography with a molecular weight of >700 kDa. Analysis by Fourier transform infrared spectroscopy and nuclear magnetic resonance (NMR) combined with acid hydrolysis to monomers showed that the polymer was built of a backbone of β(1 → 4)-linked xylose residues that were substituted by 4-O-methylglucuronic acids, arabinose, and galactose in an approximate molar ratio of 114:23:5:1. NMR analysis also indicated the presence of α(1 → 5)-linked arabinose substituents in dimeric or oligomeric forms. The main xylooligosaccharides (XOs) produced after hydrolysis of the extracted glucuronoarabinoxylan polymer by thermostable glycoside hydrolases (GHs) from families 10 and 11 were xylobiose and xylotriose, followed by peaks of putative substituted XOs. Quantification of the unsubstituted XOs using standards showed that the highest yield from the soluble glucuronoarabinoxylan fraction was 1.26 g/100 g of xylan fraction, only slightly higher than the yield (1.00 g/100 g of xylan fraction) from the insoluble fraction (p 0.05). This study shows that quinoa stalks represent a novel source of glucuronoarabinoxylan, with a substituent structure that allowed for limited production of XOs by GH10 or GH11 enzymes.

  2. ORF Alignment: NC_005090 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... [Symbiobacterium thermophilum IAM 14863] ... Length = 126 ... Query: 18 ... VLDAEGKTFGRLMTQVATLLRGKHRPDYTPHVDC...GDFVVIVNASKVVFSGLKLEAKEYFT 77 ... V+DAEGKT+GRL ++VA +LRGKH+P +TPHVDC...GDFV+IVNA K+ ... +G K+ ... K Y+ ... Sbjct: 1 ... VVDAEGKTLGRLASKVAAILRGKHKPTFTPHVDCGDFVIIVNAEKIQVTGKKVTDKVYYR 60 ...

  3. Purification and characterization of novel bi-functional GH3 family β-xylosidase/β-glucosidase from Aspergillus niger ADH-11.

    Science.gov (United States)

    Patel, Harshvadan; Kumar, Adepu Kiran; Shah, Amita

    2018-04-01

    β-Xylosidase plays an important role in xylan degradation by relieving the end product inhibition of endo-xylanase caused by xylo-oligosaccharides. β-Xylosidase has a wide range of applications in food, feed, paper and pulp, pharmaceutical industries and in bioconversion of lignocellulosic biomass. Hence, in the present study focused on purification, biochemical characterization and partial sequencing of purified β-xylosidase from xylanolytic strain Aspergillus niger ADH-11. Acetone precipitation followed by GPC using Sephacryl S-200 yielded 20.59-fold purified β-xylosidase with 58.30% recovery. SDS-PAGE analysis of purified β-xylosidase relieved a monomeric subunit with a molecular weight 120.48kDa. Kinetic parameters of purified β-xylosidase viz Km, Vmax, Kcat and catalytic efficiency were assessed. Purified β-xylosidase was additionally active on p-nitrophenyl-β-d-glucopyranoside substrate also. Moreover, peptide mass fingerprinting analysis support our biochemical studies and showed that the purified protein is a novel β-xylosidase with β-glucosidase activity and belongs to the bi-functional GH3 superfamily. Besides, tolerance of purified β-xylosidase towards glucose and xylose was also assessed. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Deletion of pH Regulator pac-3 Affects Cellulase and Xylanase Activity during Sugarcane Bagasse Degradation by Neurospora crassa.

    Directory of Open Access Journals (Sweden)

    Amanda Cristina Campos Antoniêto

    Full Text Available Microorganisms play a vital role in bioethanol production whose usage as fuel energy is increasing worldwide. The filamentous fungus Neurospora crassa synthesize and secrete the major enzymes involved in plant cell wall deconstruction. The production of cellulases and hemicellulases is known to be affected by the environmental pH; however, the regulatory mechanisms of this process are still poorly understood. In this study, we investigated the role of the pH regulator PAC-3 in N. crassa during their growth on sugarcane bagasse at different pH conditions. Our data indicate that secretion of cellulolytic enzymes is reduced in the mutant Δpac-3 at alkaline pH, whereas xylanases are positively regulated by PAC-3 in acidic (pH 5.0, neutral (pH 7.0, and alkaline (pH 10.0 medium. Gene expression profiles, evaluated by real-time qPCR, revealed that genes encoding cellulases and hemicellulases are also subject to PAC-3 control. Moreover, deletion of pac-3 affects the expression of transcription factor-encoding genes. Together, the results suggest that the regulation of holocellulase genes by PAC-3 can occur as directly as in indirect manner. Our study helps improve the understanding of holocellulolytic performance in response to PAC-3 and should thereby contribute to the better use of N. crassa in the biotechnology industry.

  5. Purification, crystallization and preliminary X-ray analysis of a thermostable glycoside hydrolase family 43 beta-xylosidase from Geobacillus thermoleovorans IT-08

    NARCIS (Netherlands)

    Rohman, Ali; van Oosterwijk, Niels; Kralj, Slavko; Dijkhuizen, Lubbert; Dijkstra, Bauke W.; Puspaningsih, Ni Nyoman Tri

    2007-01-01

    The main enzymes involved in xylan-backbone hydrolysis are endo-1,4-beta-xylanase and beta-xylosidase. beta-Xylosidase converts the xylo-oligosaccharides produced by endo-1,4-beta-xylanase into xylose monomers. The beta-xylosidase from the thermophilic Geobacillus thermoleovorans IT-08, a member of

  6. ISOLASI, SKRINING DAN IDENTIFIKASI JAMUR XILANOLITIK LOKAL YANG BERPOTENSI SEBAGAI AGENSIA PEMUTIH PULP YANG RAMAH LINGKUNGAN (Isolation, Screening and Identification Xylanolytic Local Fungi that Potentially as Pulp Bleaching Agents

    Directory of Open Access Journals (Sweden)

    Elisa Nurnawati

    2015-01-01

    Xylanase has great potential for industry application. Application of xylanase can be done in pretreatment of pulp bleaching in the pulp and paper industry. Enzyme application can reduce the use of chlorine compounds that are harmful to the environment. Therefore, xylanase that used in pulp bleaching should be free of cellulase activity. Fungi are one of the groups of microbes that are able to produce xylanase. The aims of this study was to obtain local xylanase-producing fungal isolates from soil that assumed contain of xylan. The source of fungal isolates were the soil around the pulp and paper industry; Acacia forests in the district Ogan Ilir and Muara Enim, South Sumatra; Wanagama, Yogyakarta; sawmills in Palembang and Yogyakarta; and Palembang landfill. Based on the initial screening in the agar basal medium, 111 fungal isolates were obtained. Most of them were the xylanase-producing fungi, but only 12 fungal isolates that have high xylanolytic capabilities. Further screening was performed on xylan liquid basal medium. The results showed that the fungus identified as Chaetomium globosum, Penicillium simplicissimum, Aspergillus tamarii and Monocillium have higher xylanase specific activity than the other isolates. They were also have lignolytic and cellulolytic activities. Therefore, fungal xylanase potentially developed as a pulp bleaching agent.

  7. Production and characterization of thermostable xylanase from ...

    African Journals Online (AJOL)

    ajl2

    2013-02-20

    Feb 20, 2013 ... Penicillium, .... production in liquid medium ranged from 0.11 to 0.21. U/ml (data not shown). ... Lane 2 represents the negative control (Bacillus sp.). Lane 3 is .... circulans D1 in submerged fermentation using response surface.

  8. An Alignment-Free Algorithm in Comparing the Similarity of Protein Sequences Based on Pseudo-Markov Transition Probabilities among Amino Acids.

    Science.gov (United States)

    Li, Yushuang; Song, Tian; Yang, Jiasheng; Zhang, Yi; Yang, Jialiang

    2016-01-01

    In this paper, we have proposed a novel alignment-free method for comparing the similarity of protein sequences. We first encode a protein sequence into a 440 dimensional feature vector consisting of a 400 dimensional Pseudo-Markov transition probability vector among the 20 amino acids, a 20 dimensional content ratio vector, and a 20 dimensional position ratio vector of the amino acids in the sequence. By evaluating the Euclidean distances among the representing vectors, we compare the similarity of protein sequences. We then apply this method into the ND5 dataset consisting of the ND5 protein sequences of 9 species, and the F10 and G11 datasets representing two of the xylanases containing glycoside hydrolase families, i.e., families 10 and 11. As a result, our method achieves a correlation coefficient of 0.962 with the canonical protein sequence aligner ClustalW in the ND5 dataset, much higher than those of other 5 popular alignment-free methods. In addition, we successfully separate the xylanases sequences in the F10 family and the G11 family and illustrate that the F10 family is more heat stable than the G11 family, consistent with a few previous studies. Moreover, we prove mathematically an identity equation involving the Pseudo-Markov transition probability vector and the amino acids content ratio vector.

  9. purified from a thermophilic fungus Scytalidium thermophilum 15.8 ...

    African Journals Online (AJOL)

    GRACE

    2006-06-16

    Jun 16, 2006 ... Fax: +55 16 36331758. are in the production of high glucose syrup (HGS) from ... The amount of reducing sugar released was estimated by glucose-oxidase procedure ... Buthanol/ethanol/water (5:3:2,. V/V) mix was used as a ...

  10. Towards a molecular understanding of symbiont function: Identification of a fungal gene for the degradation of xylan in the fungus gardens of leaf-cutting ants

    Directory of Open Access Journals (Sweden)

    Lange Lene

    2008-02-01

    Full Text Available Abstract Background Leaf-cutting ants live in symbiosis with a fungus that they rear for food by providing it with live plant material. Until recently the fungus' main inferred function was to make otherwise inaccessible cell wall degradation products available to the ants, but new studies have shed doubt on this idea. To provide evidence for the cell wall degrading capacity of the attine ant symbiont, we designed PCR primers from conserved regions of known xylanase genes, to be used in PCR with genomic DNA from the symbiont as template. We also measured xylanase, cellulase and proteinase activities in the fungus gardens in order to investigate the dynamics of degradation activities. Results We cloned a xylanase gene from the mutualistic fungus of Acromyrmex echinatior, determined its protein sequence, and inserted it in a yeast expression vector to confirm its substrate specificity. Our results show that the fungus has a functional xylanase gene. We also show by lab experiments in vivo that the activity of fungal xylanase and cellulase is not evenly distributed, but concentrated in the lower layer of fungus gardens, with only modest activity in the middle layer where gongylidia are produced and intermediate activity in the newly established top layer. This vertical distribution appears to be negatively correlated with the concentration of glucose, which indicates a directly regulating role of glucose, as has been found in other fungi and has been previously suggested for the ant fungal symbiont. Conclusion The mutualistic fungus of Acromyrmex echinatior has a functional xylanase gene and is thus presumably able to at least partially degrade the cell walls of leaves. This finding supports a saprotrophic origin of the fungal symbiont. The observed distribution of enzyme activity leads us to propose that leaf-substrate degradation in fungus gardens is a multi-step process comparable to normal biodegradation of organic matter in soil ecosystems

  11. Prospect for Developing a Consolidated Bioprocessing (CBP) Strain Using Xylan as the Substrate: the Case Study of Yarrowia lipolytica

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Wei; Wei, Hui; Alahuhta, Markus; Zhang, Min; Himmel, Michael E.

    2016-07-08

    To achieve the goal of developing a direct microbial sugar conversion platform for the production of lipids and drop-in fuels from cellulosic biomass substrate, Yarrowia lipolytica was used to investigate its potential for being developed as CBP strain by expressing cellulase and xylanase enzymes. Y. lipolytica is known to accumulate lipids intracellularly and is capable of metabolizing glucose and xylose to produce lipids; however, due to the lack of the biomass degrading enzymes, it cannot directly utilize lignocellulosic substrates as carbon sources. While research is continuing on the development of a Y. lipolytica strain able to degrade cellulose, in this study, we present successful expression of several xylanases in Y. lipolytica. To the best of our knowledge, this is the first study introducing heterologous hemicellulose genes into the genome of Y. lipolytica. SDS-PAGE and western blotting analysis showed that the endo-xylanase gene XynII and exo-xylosidase gene XlnD were successfully expressed and secreted, and the expressed xylanases were likely either not or sparsely glycosylated, which is advantageous for expression of heterologous proteins from any species. Enzymatic activity tests further demonstrated active expression of XynII and XlnD in Y. lipolytica. Furthermore, synergistic action on converting xylan to xylose was observed when XlnD worked in concert with XynII. XlnD was able to work on the xylo-oligomers generated by XynII, enhancing the xylan conversion to monomeric xylose. The successful expression of these xylanases in Yarrowia further advances us towards our goal to develop a direct microbial conversion process using this organism. and xylose to produce lipids; however, due to the lack of the biomass degrading enzymes, it cannot directly utilize lignocellulosic substrates as carbon sources. While research is continuing on the development of a Y. lipolytica strain able to degrade cellulose, in this study, we present successful expression of

  12. Molecular characterization of two alkylresorcylic acid synthases from Sordariomycetes fungi

    DEFF Research Database (Denmark)

    Ramakrishnan, Dhivya; Tiwari, Manish Kumar; Manoharan, Gomathi

    2018-01-01

    Two putative type III polyketide synthase genes (PKS) were identified from Sordariomycetes fungi. These two type III PKS genes from Sordaria macrospora (SmPKS) and Chaetomium thermophilum (CtPKS), shared 59.8% sequence identity. Both, full-length and truncated versions of type III PKSs were...

  13. The Mycoflora of Hot Spring Soil in Northern Taiwan

    Directory of Open Access Journals (Sweden)

    Kuei-Yu Chen

    2003-09-01

    Full Text Available An investigation of the mycoflora in northern Taiwan from August 1999 to June 2000, particularly of thermophilic and thermotolerant fungi inhabiting sulfurous hot spring soils, resulted in identification 12 taxa: Aspergillus fumigatus var. fumigatus (66.85 %, A. fumigatus var. 1 with green colony (7.86 %, A. fumigatus var. 2 with brown colony (4.81 %, A. niger (1.14 %, unidentified Asperigillus sp. (0.045 %, Chrysosporium sp. (0.18 %, Papulaspora thermophila (2.72 %, Scytalidium thermophilum (0.045 %, Sporotrichum sp. (0.045 %, Mycelia sterilia sp.1 with white colony (6.63 %, Mycelia sterilia sp.2 with yellow colony (5.27 % and Mycelia sterilia sp. 3 with gray colony (4.405 %. A total of 2202 colonies were isolated from three sampling sites: site 1 (hot springhead, site 2 (2 m from site 1 and site 3 (4 m from site 1. Fungal colonies isolated as well as species percentage at three sites were as follows: 32.92 % in 9 taxa from site 1, 37.87 % in 11 taxa from site 2, and 29.21 % in 8 taxa from site 3. The dominant species was Aspergillus fumigatus var. fumigatus, which was isolated year around from three sampling sites. A. fumigatus var. 1 appeared from February to June 2000. A. fumigatus var. 2 was isolated only in August and October 1999. Within the sampling range of hot spring niches, there was evidence of the presence of ecotypes in the A. fumigatus complex. Chrysosporium sp. and Sporotrichum sp. were isolated only from the soils without hot water treatment, but Aspergillus sp. and Scytalidium thermophilum were isolated only from the soils pre-treated with hot water for 30 min. at 60℃. The significance level (P value of fungal communities between hot water treatment and no treatment was 0.866, indicating that no significant difference between both treatments.

  14. A simple, versatile, low-cost and remotely operated apparatus for [11C]acetate, [11C]choline, [11C]methionine and [11C]PIB synthesis

    International Nuclear Information System (INIS)

    Cheung Manki; Ho Chilai

    2009-01-01

    A simple, efficient and remotely operated synthesis apparatus for carrying out routine [ 11 C]carboxylation, on-column and bubbling [ 11 C]methylation was essential for reliable, day-to-day production of [ 11 C]-labelled PET radiopharmaceuticals. We developed an in-house apparatus specifically applied to the synthesis of [ 11 C]acetate, [ 11 C]choline, [ 11 C]methionine and 2-(4'-N-[ 11 C]methylaminophenyl)-6-hydroxybenzothiazole ([ 11 C]PIB), where high radiochemical purity (≥97%) and moderate radiochemical yields (18% for [ 11 C]PIB, 41-55% for the others) could be achieved. These findings provided evidence that this was a fast, versatile and reliable apparatus suitable for a PET/CT centre with limited financial budget and hot cell space for synthesis of [ 11 C]-labelled radiopharmaceuticals

  15. Evaluation of gastrointestinal bacterial population for the production of holocellulose enzymes for biomass deconstruction.

    Science.gov (United States)

    Asem, Dhaneshwaree; Leo, Vincent Vineeth; Passari, Ajit Kumar; Tonsing, Mary Vanlalhruaii; Joshi, J Beslin; Uthandi, Sivakumar; Hashem, Abeer; Abd Allah, Elsayed Fathi; Singh, Bhim Pratap

    2017-01-01

    The gastrointestinal (GI) habitat of ruminant and non-ruminant animals sustains a vast ensemble of microbes that are capable of utilizing lignocellulosic plant biomass. In this study, an indigenous swine (Zovawk) and a domesticated goat (Black Bengal) were investigated to isolate bacteria having plant biomass degrading enzymes. After screening and enzymatic quantification of eighty-one obtained bacterial isolates, Serratia rubidaea strain DBT4 and Aneurinibacillus aneurinilyticus strain DBT87 were revealed as the most potent strains, showing both cellulase and xylanase production. A biomass utilization study showed that submerged fermentation (SmF) of D2 (alkaline pretreated pulpy biomass) using strain DBT4 resulted in the most efficient biomass deconstruction with maximum xylanase (11.98 U/mL) and FPase (0.5 U/mL) activities (55°C, pH 8). The present study demonstrated that bacterial strains residing in the gastrointestinal region of non-ruminant swine are a promising source for lignocellulose degrading microorganisms that could be used for biomass conversion.

  16. Evaluation of gastrointestinal bacterial population for the production of holocellulose enzymes for biomass deconstruction.

    Directory of Open Access Journals (Sweden)

    Dhaneshwaree Asem

    Full Text Available The gastrointestinal (GI habitat of ruminant and non-ruminant animals sustains a vast ensemble of microbes that are capable of utilizing lignocellulosic plant biomass. In this study, an indigenous swine (Zovawk and a domesticated goat (Black Bengal were investigated to isolate bacteria having plant biomass degrading enzymes. After screening and enzymatic quantification of eighty-one obtained bacterial isolates, Serratia rubidaea strain DBT4 and Aneurinibacillus aneurinilyticus strain DBT87 were revealed as the most potent strains, showing both cellulase and xylanase production. A biomass utilization study showed that submerged fermentation (SmF of D2 (alkaline pretreated pulpy biomass using strain DBT4 resulted in the most efficient biomass deconstruction with maximum xylanase (11.98 U/mL and FPase (0.5 U/mL activities (55°C, pH 8. The present study demonstrated that bacterial strains residing in the gastrointestinal region of non-ruminant swine are a promising source for lignocellulose degrading microorganisms that could be used for biomass conversion.

  17. Direct production of feruloyl oligosaccharides and hemicellulase inducement and distribution in a newly isolated Aureobasidium pullulans strain.

    Science.gov (United States)

    Yu, Xiao-hong; Gu, Zhen-xin

    2014-02-01

    Studies were carried out to screen and identify strains that are able to directly produce ferulic oligosaccharides (FOs) from wheat bran (WB). The inducement and distribution of hemicellulases from strain 2012, which was identified as a non-melanin secreting strain of Aureobasidium pullulans (A. pullulans), were also determined. In a 60 g/L WB solution, A. pullulans could produce 545 nmol/L FOs, 64.12 IU/mL xylanase and 0.14 IU/mL ferulic acid esterase (FAE). A. pullulans was cultivated in media with WB, glucose, xylose, sucrose, lactose or xylan as the carbon source, and hemicellulases were mainly induced by xylan and WB and inhibited by glucose and sucrose. Xylanase and FAE were mainly present in the culture filtrate, xylosidase in the hyphal filaments and arabinofuranosidase was a membrane-bound enzyme. The yield of FOs was positively correlated to the hemicellulases activity, and significantly positively (P < 0.05) correlated to the xylanase activity (r = 0.992).

  18. The community dynamics of major bioleaching microorganisms during chalcopyrite leaching under the effect of organics.

    Science.gov (United States)

    Li, Qihou; Tian, Ye; Fu, Xian; Yin, Huaqun; Zhou, Zhijun; Liang, Yiting; Qiu, Guanzhou; Liu, Jie; Liu, Hongwei; Liang, Yili; Shen, Li; Cong, Jing; Liu, Xueduan

    2011-08-01

    To determine the effect of organics (yeast extract) on microbial community during chalcopyrite bioleaching at different temperature, real-time polymerase chain reaction (PCR) was employed to analyze community dynamics of major bacteria applied in bioleaching. The results showed that yeast extract exerted great impact on microbial community, and therefore influencing bioleaching rate. To be specific, yeast extract was adverse to this bioleaching process at 30°C due to decreased proportion of important chemolithotrophs such as Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans. However, yeast extract could promote bioleaching rate at 40°C on account of the increased number and enhanced work of Ferroplasma thermophilum, a kind of facultative bacteria. Similarly, bioleaching rate was enhanced under the effect of yeast extract at 50°C owing to the work of Acidianus brierleyi. At 60°C, bioleaching rate was close to 100% and temperature was the dominant factor determining bioleaching rate. Interestingly, the existence of yeast extract greatly enhanced the relative competitiveness of Ferroplasma thermophilum in this complex bioleaching microbial community.

  19. Screening and characterizing of xylanolytic and xylose-fermenting yeasts isolated from the wood-feeding termite, Reticulitermes chinensis.

    Directory of Open Access Journals (Sweden)

    Sameh Samir Ali

    Full Text Available The effective fermentation of xylose remains an intractable challenge in bioethanol industry. The relevant xylanase enzyme is also in a high demand from industry for several biotechnological applications that inevitably in recent times led to many efforts for screening some novel microorganisms for better xylanase production and fermentation performance. Recently, it seems that wood-feeding termites can truly be considered as highly efficient natural bioreactors. The highly specialized gut systems of such insects are not yet fully realized, particularly, in xylose fermentation and xylanase production to advance industrial bioethanol technology as well as industrial applications of xylanases. A total of 92 strains from 18 yeast species were successfully isolated and identified from the gut of wood-feeding termite, Reticulitermes chinensis. Of these yeasts and strains, seven were identified for new species: Candida gotoi, Candida pseudorhagii, Hamamotoa lignophila, Meyerozyma guilliermondii, Sugiyamaella sp.1, Sugiyamaella sp. 2, and Sugiyamaella sp.3. Based on the phylogenetic and phenotypic characterization, the type strain of C. pseudorhagii sp. nov., which was originally designated strain SSA-1542T, was the most frequently occurred yeast from termite gut samples, showed the highly xylanolytic activity as well as D-xylose fermentation. The highest xylanase activity was recorded as 1.73 and 0.98 U/mL with xylan or D-xylose substrate, respectively, from SSA-1542T. Among xylanase-producing yeasts, four novel species were identified as D-xylose-fermenting yeasts, where the yeast, C. pseudorhagii SSA-1542T, showed the highest ethanol yield (0.31 g/g, ethanol productivity (0.31 g/L·h, and its fermentation efficiency (60.7% in 48 h. Clearly, the symbiotic yeasts isolated from termite guts have demonstrated a competitive capability to produce xylanase and ferment xylose, suggesting that the wood-feeding termite gut is a promising reservoir for novel

  20. Feruloyl esterase from Aspergillus clavatus improves xylan hydrolysis of sugarcane bagasse

    Directory of Open Access Journals (Sweden)

    Dyoni M. de Oliveira

    2016-12-01

    Full Text Available Feruloyl esterase is a subclass of carboxylic acid esterases with the capacity to release ferulic acid and other cinnamic acids from plant cell walls and synthetic substrates. Feruloyl esterases act synergistically with xylanases removing ferulic acid residues esterified to arabinoxylans. Feruloyl esterase type D from Aspergillus clavatus (AcFAE was expressed in Escherichia coli, purified, and applied with a commercial xylanase consortium (Novozymes for hydrolysis of sugarcane bagasse. Feruloyl esterase plus xylanase increased 5.13-fold the releasing of ferulic acid from sugarcane bagasse. Removal of only 7.7% of ferulic acid content by AcFAE increased 97.3% the sugarcane bagasse hydrolysis by xylanase. These data support the use of AcFAE as an interesting adjuvant enzyme to improve lignocellulose digestion and biotechnological tool for biorefineries.

  1. 26 CFR 11.401(a)-11 - Qualified joint and survivor annuities.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 14 2010-04-01 2010-04-01 false Qualified joint and survivor annuities. 11.401... SECURITY ACT OF 1974 § 11.401(a)-11 Qualified joint and survivor annuities. (a) In general—(1) General rule... annuity (i.e., an annuity requiring survival of the participant or his spouse as a condition for payment...

  2. A Moderately Thermophilic Mixed Microbial Culture for Bioleaching of Chalcopyrite Concentrate at High Pulp Density

    Science.gov (United States)

    Wang, Yuguang; Zeng, Weimin; Qiu, Guanzhou; Chen, Xinhua

    2014-01-01

    Three kinds of samples (acid mine drainage, coal mine wastewater, and thermal spring) derived from different sites were collected in China. Thereafter, these samples were combined and then inoculated into a basal salts solution in which different substrates (ferrous sulfate, elemental sulfur, and chalcopyrite) served as energy sources. After that, the mixed cultures growing on different substrates were pooled equally, resulting in a final mixed culture. After being adapted to gradually increasing pulp densities of chalcopyrite concentrate by serial subculturing for more than 2 years, the final culture was able to efficiently leach the chalcopyrite at a pulp density of 20% (wt/vol). At that pulp density, the culture extracted 60.4% of copper from the chalcopyrite in 25 days. The bacterial and archaeal diversities during adaptation were analyzed by denaturing gradient gel electrophoresis and constructing clone libraries of the 16S rRNA gene. The results show that the culture consisted mainly of four species, including Leptospirillum ferriphilum, Acidithiobacillus caldus, Sulfobacillus acidophilus, and Ferroplasma thermophilum, before adapting to a pulp density of 4%. However, L. ferriphilum could not be detected when the pulp density was greater than 4%. Real-time quantitative PCR was employed to monitor the microbial dynamics during bioleaching at a pulp density of 20%. The results show that A. caldus was the predominant species in the initial stage, while S. acidophilus rather than A. caldus became the predominant species in the middle stage. F. thermophilum accounted for the greatest proportion in the final stage. PMID:24242252

  3. InXy and SeXy, compact heterologous reporter proteins for mammalian cells.

    Science.gov (United States)

    Fluri, David A; Kelm, Jens M; Lesage, Guillaume; Baba, Marie Daoud-El; Fussenegger, Martin

    2007-10-15

    Mammalian reporter proteins are essential for gene-function analysis, drugscreening initiatives and as model product proteins for biopharmaceutical manufacturing. Bacillus subtilis can maintain its metabolism by secreting Xylanase A (XynA), which converts xylan into shorter xylose oligosaccharides. XynA is a family 11 xylanase monospecific for D-xylose containing substrates. Mammalian cells transgenic for constitutive expression of wild-type xynA showed substantial secretion of this prokaryotic enzyme. Deletion analysis confirmed that a prokaryotic signal sequence encoded within the first 81 nucleotides was compatible with the secretory pathway of mammalian cells. Codon optimization combined with elimination of the prokaryotic signal sequence resulted in an exclusively intracellular mammalian Xylanase A variant (InXy) while replacement by an immunoglobulin-derived secretion signal created an optimal secreted Xylanase A derivative (SeXy). A variety of chromogenic and fluorescence-based assays adapted for use with mammalian cells detected InXy and SeXy with high sensitivity and showed that both reporter proteins resisted repeated freeze/thaw cycles, remained active over wide temperature and pH ranges, were extremely stable in human serum stored at room temperature and could independently be quantified in samples also containing other prominent reporter proteins such as the human placental alkaline phosphatase (SEAP) and the Bacillus stearothermophilus-derived secreted alpha-amylase (SAMY). Glycoprofiling revealed that SeXy produced in mammalian cells was N- glycosylated at four different sites, mutation of which resulted in impaired secretion. SeXy was successfully expressed in a variety of mammalian cell lines and primary cells following transient transfection and transduction with adeno-associated virus particles (AAV) engineered for constitutive SeXy expression. Intramuscular injection of transgenic AAVs into mice showed significant SeXy levels in the bloodstream

  4. PDP-11 front-end for a VAX-11/780

    International Nuclear Information System (INIS)

    Browne, M.J.; Granieri, C.; Sherden, D.J.; Weaver, L.J.

    1980-01-01

    An unpublicized feature of the VAX-11/780 is the provision for attaching a PDP-11 to the VAX UNIBUS Adapter. Doing this can give significantly improved I/O performance for applications which are limited by overhead in the VAX I/O driver rather than by the transfer speed of the UNIBUS itself. Such a system was implemented by using a PDP-11/04 as a front-end to a CAMAC data acquisition system. Both the PDP and the VAX have full access to the UNIBUS. That portion of the PDP address space that does not have UNIBUS memory can be mapped to buffers in the VAX memory; this approach allows the PDP to access VAX memory and to initiate DMA transfers directly to the VAX. The VAX also has full access to the PDP memory; a convenient means for developing and downloading the PDP software is thus provided. 5 figures

  5. A study of 11C-acetate production using 11C-choline commercial module

    International Nuclear Information System (INIS)

    Zhang Jinming; Tian Jiahe; Yao Shulin; Chen Yan

    2008-01-01

    Objective: 11 C-acetate is a useful PET tracer in evaluating myocardial metabolism but more interest has been focused on its application in tumor detection in recent years, especially hepatocellular carcinoma (HCC). This study was designed to evaluate the laboratory, preparation of 11 C-acetate with a modified 11 C-choline commercial module and to investigate its biodistribution in tumor (lung adenocarcinoma)-bearing mice as well as its potential as a tumor imaging agent. Methods: 11 C-acetate was synthesized with a modified 11 C choline module: Methyl magnesium bromide Grignard (0.1 ml of 1.5 mol/L) was load- ed to a Teflon loop before 11 CO 2 was recovered from the target. Cartier acetate solution (2 ml of 1 mmol/L) was pushed through the loop, SPE cartridges (mixed AG50 and IC-Ag) and then the QMA. The loop and cartridges were then rinsed with water. The product 11 C-acetate was then washed out from QMA with 0.9% NaCl solution into a collection vial containing diluted HCl. 11 C-carbonate was removed by nitrogen bubbling for 2 min. After neutralization with trisodium citrate, the injectable 11 C-acetate solution was obtained. The tumor-bearing mice were sacrificed. The percentage activity of injected dose per gram of tissue (% ID/g) for tumor and other tissues were calculated. One patient with known diagnosis of moderately differentiated HCC was injected with 11 C-acetate and imaged by PET/CT, followed by 18 F-fluorodeoxyglucose (FDG). Results: The synthesis yield of 11 C-acetate was (60.5 ± 8.7)% (decay conected, n=10); the radio-chemistry purity was > 98% and the synthesis time was 10 min from 11 CO 2 to 11 C-acetate. The radioactivity ratio for tumor/muscle was 1.76 at 30 min. A patient with known HCC showed positive 11 C-acetate accumulation in the tumor but was negative in 18 F-FDG. Conclusion: The synthesis of 11 C-acetate by modification of an 11 C-choline commercial module was feasible and it could be achieved with high yield, high radiochemical purity

  6. Characterization of a Rab11-like GTPase, EhRab11, of Entamoeba histolytica.

    Science.gov (United States)

    McGugan, Glen C; Temesvari, Lesly A

    2003-07-01

    The Entamoeba histolytica Rab11 family of small molecular weight GTPases consists of three members, EhRab11, EhRab11B, and EhRab11C. The functions of these Rabs in Entamoeba have not been determined. Therefore, as an approach to elucidate the role of the Rab11 family of GTPases in Entamoeba, immunofluorescence microscopy was undertaken to define the subcellular localization of one member of this family, EhRab11. Under conditions of growth, EhRab11 displayed a punctate pattern in the cytoplasm of trophozoites. EhRab11 did not colocalize with markers for the Golgi apparatus, endoplasmic reticulum, pinosomes, phagosomes, or compartments formed by receptor-mediated endocytosis, suggesting that this Rab may not play a role in vesicle trafficking between these organelles. Under conditions of iron and serum starvation, EhRab11 was translocated to the periphery of the cell. The altered cellular localization was accompanied by multinucleation of the cells as well as the acquisition of detergent resistance by the cells, features that are characteristic of Entamoeba cysts. The translocation of EhRab11 to the periphery of the cell during iron and serum starvation was specific as the subcellular localizations of two other Rab GTPases, EhRab7 and EhRabA, were not altered under the same conditions. In addition, the formation of multinucleated cells by inhibition of cytokinesis was not sufficient to induce the translocation of EhRab11 to the cell periphery. Taken together, the data suggest that iron and serum starvation may induce encystation in E. histolytica and that EhRab11 may play a role in this process. Moreover, these studies are the first to describe a putative role for a Rab GTPase in encystation in Entamoeba sp.

  7. Fermentation characteristics in hay from Cynodon and crop stubble treated with exogenous enzymes

    Directory of Open Access Journals (Sweden)

    Yânez André Gomes Santana

    Full Text Available ABSTRACT The effect of treatment with xylanase and β-glucanase was evaluated for gas production and the ruminal degradation of nutrients from the hay of Tifton 85 grass and the stubble of maize, sorghum, peanut, sunflower and sesame crops. Two commercial fibrolytic enzymes were used (Dyadic xylanase PLUS - Xylanase; BrewZyme LP-β-glucanase, added to the hay at doses of 7.5 units of endoglucanase and 0.46 units of xylanase per 500 mg/gDM, for the cellulase and xylanase products respectively. The chemical composition of the hay was determined for no enzyme application and 24 hours after enzyme treatment, and the in vitro gas production and in situ microbial degradation was estimated for dry matter, organic matter, neutral detergent fibre and truly-degradable organic matter after 24 hours of incubation in the rumen. Enzyme treatment of the hay from Tifton 85 grass and the stubble of maize, sorghum, sunflower, peanut and sesame crops with the exogenous fibrolytic enzymes β-glucanase and xylanase influences in vitro gas production, and the in situ degradation of dry matter, organic matter, neutral detergent fibre and truly-degradable organic matter in the rumen. This variation can be attributed to differences in the chemical composition of the hay from the grass and the crop stubble, and to the different ways the enzymes act upon the cell wall.

  8. Isolation, cloning and molecular characterization of a thermotolerant ...

    African Journals Online (AJOL)

    Isolation, cloning and molecular characterization of a thermotolerant xylanase from Streptomyces sp. THW31. Thayat Sriyapai, Peechapack Somyoonsap, Supatra Areekit, Paisarn Khawsak, Arda Pakpitcharoen, Kosum Chansiri ...

  9. Efeitos do centeio, do trigo e da suplementação com xilanases sobre o valor nutricional de dietas e o desempenho de frangos corte Effects of rye, wheat and xylanase supplementation on diet nutritive value and broiler chicken performance

    Directory of Open Access Journals (Sweden)

    José Luís Teixeira de Abreu Medeiros Mourão

    2009-12-01

    Full Text Available Três experimentos foram realizados para avaliar os efeitos de dietas formuladas com 53% de trigo ou centeio, suplementadas ou não com a enzima xilanase (0,06%, sobre a digestibilidade de nutrientes e a energia metabolizável das dietas e sobre o desempenho e desenvolvimento do trato digestivo de frangos de corte. Como testemunha foi usada uma dieta com 53% de milho. No primeiro experimento, as digestibilidades da matéria seca, matéria orgânica e gordura e energia metabolizável corrigida para o nitrogênio (EMAn das dietas com centeio foram menores que daquelas com trigo (2.556 kcal/kg vs 2.842 kcal/kg e menores no conjunto dessas dietas que na dieta com milho (2.684 kcal/kg vs 3.010 kcal/kg. A digestibilidade da matéria orgânica e a EMAn das dietas com centeio também foi inferior às da dieta com trigo. A suplementação com xilanases não afetou a utilização digestiva das dietas com centeio ou com trigo. No segundo experimento, nos frangos alimentados com as dietas com centeio, o menor ganho de peso e consumo de EMAn e a pior conversão alimentar ocorreram entre os 8 e os 35 dias de idade. A conversão alimentar foi de 2,17 para as dietas com centeio e 1,88 para a dieta com milho. A adição de xilanases às dietas com centeio ou trigo não melhorou a conversão alimentar. Nos frangos alimentados com as dietas com centeio durante 31 dias, o duodeno-jejuno e íleo foram maiores e o rendimento de carcaça menor que nos frangos alimentados com a dieta testemunha (71,6% vs 74,4%. As dietas com trigo não afetaram essas características. Os pesos do pâncreas e do fígado também não foram alterados pelas dietas.Three trials with were conducted to evaluate the effects of diets with 53% rye (diet C or 53% wheat (diet T supplemented or not with xylanase enzymes (0.06% on nutrient digestibility and metabolizable energy and the performance and development of the broiler chicken digestive tract. A diet with 53% corn was used as control. In

  10. What makes organic livestock production sustainable?

    DEFF Research Database (Denmark)

    Oudshoorn, Frank W; Stubsgaard, A; Grøn Sørensen, Claus

    2014-01-01

    The effect of a current commercial xylanase (DAN) and experimental xylanase (EX), and EX in combination with protease (EXP), on the degradation and apparent ileal digestibility (AID) of non-starch polysaccharides (NSP) in wheat Distillers Dried Grains with Solubles (DDGS), was studied in 8 ileum-...

  11. Effects of cell wall degrading enzymes on carbohydrate fractions and metabolites in stomach and ileum of pigs fed wheat bran based diets

    NARCIS (Netherlands)

    Meulen, van der J.; Inborr, J.; Bakker, J.G.M.

    2001-01-01

    Pigs were fed diets containing 40 heat bran incubated with a water:acetic acid mixture (control, C) and a cellulase (Cel-i) or xylanase (Xyl-i) preparation or with addition of the cellulase (Cel-a) or xylanase (Xyl-a) preparation immediately before feeding. Stomach and ileal samples were analysed

  12. Semi-automated preparation of a 11C-labelled antibiotic - [N-methyl-11C]erythromycin A lactobionate

    International Nuclear Information System (INIS)

    Pike, V.W.; Palmer, A.J.; Horlock, P.L.; Liss, R.H.

    1984-01-01

    A fast semi-automated method is described for labelling the antibiotic, erythromycin A (1), with the short-lived positron-emitting radionuclide, 11 C(tsub(1/2)=20.4 min), in order to permit the non-invasive study of its tissue uptake in vivo. Labelling was achieved by the fast reductive methylation of N-demethylerythromycin A (2) with [ 11 C]formaldehyde, itself prepared from cyclotron-produced [ 11 C]-carbon dioxide. Rapid chemical and radiochemical purification of the [N-methyl- 11 C]erythromycin A (3) were achieved by HPLC and verified by TLC with autoradiography. The purified material was formulated for human i.v. injection as a sterile apyrogenic solution of the lactobionate salt. The preparation takes 42 min from the end of radionuclide production and from [ 11 C]carbon dioxide produces [N-methyl- 11 C]erythromycin A lactobionate in 4-12% radiochemical yield, corrected for radioactive decay. (author)

  13. Next-generation non-starch polysaccharide-degrading, multi-carbohydrase complex rich in xylanase and arabinofuranosidase to enhance broiler feed digestibility.

    Science.gov (United States)

    Cozannet, Pierre; Kidd, Michael T; Montanhini Neto, Roberto; Geraert, Pierre-André

    2017-08-01

    This study was carried out to evaluate the effect of a multi-carbohydrase complex (MCC) rich in xylanase (Xyl) and arabinofuranosidase (Abf) on overall broiler feed digestibility in broilers. Energy utilization and digestibility of dry matter (DM), organic matter (OM), protein, starch, fat, and insoluble and soluble fibers were measured using the mass-balance method. The experiment was carried out on 120 broilers (3-week-old chickens). Broilers were distributed over 8 treatments to evaluate the effect of the dietary arabinoxylan content and nutrient density with and without MCC (Rovabio® Advance). The graded content of arabinoxylan (AX) was obtained using different raw materials (wheat, rye, barley, and dried distillers' wheat). Diet-energy density was modified with added fat. Measurements indicated that nutrient density and AX content had a significant effect on most digestibility parameters. Apparent metabolizable energy (AME) was significantly increased (265 kcal kg-1) by MCC. The addition of MCC also resulted in significant improvement in the digestibility of all evaluated nutrients, with average improvements of 3.0, 3.3, 3.2, 3.0, 6.2, 2.9, 5.8, and 3.8% units for DM, OM, protein, starch, fat, insoluble and soluble fibers, and energy utilization, respectively. The interaction between MCC and diet composition was significant for the digestibility of OM, fat, protein, and energy. Nutrient digestibility and diet AME were negatively correlated with AX content (P digestible nutrient (i.e., starch, protein, fat, insoluble and soluble fibers) content with and without MCC (R2 = 0.87; RSD = 78 kcal kg-1). This study confirms that the presence of AX in wheat-based diets and wheat-based diets with other cereals and cereal by-products reduces nutrient digestibility in broiler chickens. Furthermore, the dietary addition of MCC, which is rich in Xyn and Abf, reduced deleterious effect of fiber and improved overall nutrient digestibility in broiler diets. © 2017 Poultry

  14. Effects of wheat cultivar, metabolizable energy level, and xylanase supplementation to laying hens diet on performance, egg quality traits, and selected blood parameters

    Directory of Open Access Journals (Sweden)

    Masoud Mirzaee

    2014-10-01

    Full Text Available A 2 x 2 x 2 factorial arrangement of treatments was conducted to evaluate the effects of two dietary apparent metabolizable energy (AME levels (2,720 and 2,580 kcal kg-1 diet and enzyme (0 and 0.3 g kg-1 diet, Grindazym® GP 15,000 with mostly xylanase activity supplementation on the performance of laying hens fed diets based on two wheat cultivars (Marvdasht and Sardari. Experimental diets were formulated to have a constant energy to protein ratio and were fed to 65-wk-old Lohmann LSL-Lite laying hens for 7 wk. The lower level of AME reduced egg production and egg mass (p<0.05 and increased feed conversion ratio (p<0.05. Enzyme addition increased feed intake of the birds fed a diet with Sardari cultivar (p<0.05 but had no effect on feed intake of the birds fed a diet with Marvdasht cultivar (p>0.05. Nevertheless, birds receiving diets based on Marvdasht cultivar had higher feed intake and egg mass than that of those receiving diets based on Sardari cultivar (p<0.05. The birds fed diets based on Marvdasht cultivar produced less undesired eggs and had better yolk color as compared with the birds fed diets based on Sardari cultivar (p<0.05. The serum concentration of glucose increased by enzyme supplementation when birds receiving lower AME level (p<0.05. These results indicate that enzyme supplementation may have a positive effect on the feed intake of laying hens when fed on wheat-based diets; however, this effect is cultivar dependent and does not necessarily mean that enzyme supplementation always benefit production.

  15. S-protected thiolated hydroxyethyl cellulose (HEC): Novel mucoadhesive excipient with improved stability.

    Science.gov (United States)

    Leonaviciute, Gintare; Bonengel, Sonja; Mahmood, Arshad; Ahmad Idrees, Muneeb; Bernkop-Schnürch, Andreas

    2016-06-25

    The aim of this study was the design of novel S-protected thiolated hydroxyethyl cellulose (HEC) and the assessment of its mucoadhesive properties and biodegradability compared to the corresponding unmodified polymer. Thiolated HEC was S-protected via disulfide bond formation between 6-mercaptonicotinamide (6-MNA) and the thiol substructures of the polymer. In vitro screening of mucoadhesive properties was accomplished using two different methods: rotating cylinder studies and viscosity measurements. Moreover, biodegradability of these polymers by cellulase, xylanase and lysozyme was evaluated. MTT and LDH assays were performed on Caco-2 cells to determine the cytotoxicity of S-protected thiolated HEC. Thiolated HEC displayed 280.09±1.70μmol of free thiol groups per gram polymer. S-protected thiolated HEC exhibiting 270.8±21.11μmol immobilized 6-MNA ligands per gram of polymer was shown being 2.4-fold more mucoadhesive compared to thiolated HEC. No mucoadhesion was observed in case of unmodified HEC. Results were in a good agreement with rheological studies. The presence of free thiol moieties likely caused lower degree of hydrolysis by xylanase, whereas the degradation by both enzymes cellulase and xylanase was more hampered when 6-MNA was introduced as ligand for thiol group's protection. Findings in cell viability revealed that all three conjugates were non-toxic. S-protection of thiolated hydroxyethyl cellulose improved mucoadhesive properties and provided pronounced stability towards enzymatic attack, that makes this excipient superior for non-invasive drug administration over thiolated and unmodified forms. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Two-stage in vitro digestibility assay, a tool for formulating non-starch polysaccharide degrading enzyme combinations for commonly used feed ingredients of poultry rations

    Directory of Open Access Journals (Sweden)

    Y. Ramana Reddy

    2013-05-01

    Full Text Available Aim: An attempt was made to assess the effect of pure enzyme combinations with the objective of formulating customized enzyme mixtures based on sugar release when subjected to two-stage in vitro digestion assay. Materials and Methods: A two-stage in vitro digestibility assay was carried out for commonly used feed ingredients for poultry viz., maize, soy bean meal, sunflower cake, and de-oiled rice bran supplemented with three concentrations of xylanase (5000; 7500 and 10000 IU/kg, cellulase (50; 100 and 400 IU/kg and â-D-glucanase (100; 200 and 400 IU/kg were used to formulate various NSP enzymes combinations. In total 27 NSP enzyme combinations (3x3x3 were formulated and the sugar released due to NSP digestion was quantified by phenol sulphuric acid method. Results: The total sugar release was significantly (P<0.05 higher with supplementation of various enzymes combinations for maize, sunflower cake and de-oiled rice bran where as no significant (P<0.05 interaction of various NSP enzymes combinations was observed for soy bean meal. The NSP digestibility was highest in combination (xylanase-5000, cellulase-50 and â-D-glucanase-400 IU/kg, (xylanase-10000, cellulase-50 and â-D-glucanase-200 IU/kg and (xylanase-7500, cellulase- 100 and â-D-glucanase-100 IU/kg for maize, sunflower cake and de-oiled rice bran respectively. In case of sunflower cake, significant (P<0.01 three way interaction was observed among the xylanase, cellulose, and â-D-glucanase enzymes and the two-way interactions between the enzymes were also significant (P<0.01. Conclusion: It is concluded that 'n' number of non-starch Polysaccharide enzymes combinations can be screened for their efficiency to digest non-starch Polysaccharides present in various feed ingredients commonly used in poultry rations by employing two-stage in vitro digestibility assay as a tool. [Vet World 2013; 6(8.000: 525-529

  17. Strain Improvement of Fungi by Induced Mutation through Gamma Irradiation and Selection for Animal Feed Enzymes Production and its Fermentation Process

    International Nuclear Information System (INIS)

    Konsue, Parichart; Piadang, Nattayana; Kitpreechavanich, Vichien

    2006-09-01

    Ten from eighty-nine strains of thermophilic fungi Thermomyces lanuginosus produced high level insoluble xylan degrading enzyme when cultured in submerge condition using untreated corncob as a substrate. Strain of T. lanuginosus THKU56 produced high level of insoluble xylan degrading enzyme with the most stable which was remained 28.2 and 58.9 % after treated at pH 3.5 and 70 o C for 1 h, respectively. To improve xylanase production, the strain was subjected to mutate using gamma ray at 0.4 - 1.6 kGy. The result showed the mutant strains produced insoluble xylanase activity lesser than wild type. Thus wild type strain THKU56 was then selected as potent strains for enzyme production and medium optimization was investigated using a central composite design. The four components, corncobs, yeast extract, KH 2 PO 4 and Tween 8 0, were parameters of this study. It was found that corncobs and yeast extract were discovered to affect on the xylanase production. The optimal concentration of the active nutrients for xylanase production were 41 g/l of corncobs and 24 g/l of yeast extract, which gave a predicted yield of 526.7 units/ml after 5 days culture at a temperature of 50 o C. The xylanase activity obtained from the experiment was 541 units/ml that was close to the predicted value

  18. Ancestral genetic diversity associated with the rapid spread of stress-tolerant coral symbionts in response to Holocene climate change

    KAUST Repository

    Hume, Benjamin C. C.

    2016-04-05

    Coral communities in the Persian/Arabian Gulf (PAG) withstand unusually high salinity levels and regular summer temperature maxima of up to ∼35 °C that kill conspecifics elsewhere. Due to the recent formation of the PAG and its subsequent shift to a hot climate, these corals have had only <6, 000 y to adapt to these extreme conditions and can therefore inform on how coral reefs may respond to global warming. One key to coral survival in the world\\'s warmest reefs are symbioses with a newly discovered alga, Symbiodinium thermophilum. Currently, it is unknown whether this symbiont originated elsewhere or emerged from unexpectedly fast evolution catalyzed by the extreme environment. Analyzing genetic diversity of symbiotic algae across >5, 000 km of the PAG, the Gulf of Oman, and the Red Sea coastline, we show that S. thermophilum is a member of a highly diverse, ancient group of symbionts cryptically distributed outside the PAG. We argue that the adjustment to temperature extremes by PAG corals was facilitated by the positive selection of preadapted symbionts. Our findings suggest that maintaining the largest possible pool of potentially stress-tolerant genotypes by protecting existing biodiversity is crucial to promote rapid adaptation to present-day climate change, not only for coral reefs, but for ecosystems in general.

  19. 7 CFR 201.11a - Hybrid.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Hybrid. 201.11a Section 201.11a Agriculture... REGULATIONS Labeling Agricultural Seeds § 201.11a Hybrid. If any one kind or kind and variety of seed present in excess of 5 percent is “hybrid” seed, it shall be designated “hybrid” on the label. The percentage...

  20. Biochemical characterization of thermophilic lignocellulose degrading enzymes and their potential for biomass bioprocessing

    Energy Technology Data Exchange (ETDEWEB)

    Zambare, Vasudeo; Zambare, Archana; Christopher, Lew P. [Center for Bioprocessing Research & Development, South Dakota School of Mines and Technology, Rapid City 57701, SD (United States); Muthukumarappan, Kasiviswanath [Center for Bioprocessing Research & Development, South Dakota State University, Brookings 57007, SD (United States)

    2011-07-01

    A thermophilic microbial consortium (TMC) producing hydrolytic (cellulolytic and xylanolytic) enzymes was isolated from yard waste compost following enrichment with carboxymethyl cellulose and birchwood xylan. When grown on 5% lignocellulosic substrates (corn stover and prairie cord grass) at 60C, the thermophilic consortium produced more xylanase (up to 489 U/l on corn stover) than cellulase activity (up to 367 U/l on prairie cord grass). Except for the carboxymethyl cellulose-enriched consortium, thermo-mechanical extrusion pretreatment of these substrates had a positive effect on both activities with up to 13% and 21% increase in the xylanase and cellulase production, respectively. The optimum temperatures of the crude cellulase and xylanase were 60C and 70C with half-lives of 15 h and 18 h, respectively, suggesting higher thermostability for the TMC xylanase. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the crude enzyme exhibited protein bands of 25-77 kDa with multiple enzyme activities containing 3 cellulases and 3 xylanases. The substrate specificity declined in the following descending order: avicel>birchwood xylan>microcrystalline cellulose>filter paper>pine wood saw dust>carboxymethyl cellulose. The crude enzyme was 77% more active on insoluble than soluble cellulose. The Km and Vmax values were 36.49 mg/ml and 2.98 U/mg protein on avicel (cellulase), and 22.25 mg/ml and 2.09 U/mg protein, on birchwood xylan (xylanase). A total of 50 TMC isolates were screened for cellulase and xylanase secretion on agar plates. All single isolates showed significantly lower enzyme activities when compared to the thermophilic consortia. This is indicative of the strong synergistic interactions that exist within the thermophilic microbial consortium and enhance its hydrolytic capabilities. It was further demonstrated that the thermostable enzyme-generated lignocellulosic hydrolyzates can be fermented to bioethanol by a recombinant strain of Escherichia coli

  1. Production of hydrolytic enzymes by the plant pathogenic fungus Myrothecium verrucaria in submerged cultures Produção de enzimas hidrolíticas pelo fungo fitopatogênico Myrothecium verrucaria em culturas submersas

    Directory of Open Access Journals (Sweden)

    Fabiana Guillen Moreira

    2005-03-01

    Full Text Available The capability of the plant pathogenic fungus Myrothecium verrucaria to produce extracellular hydrolytic enzymes in submerged cultures was studied using several substrates. The fungus was able to produce different depolymerases and glycosidases, being xylanase, pectinase and protease the most important. Lipase was found in cultures developed in the presence of olive oil, while protease activity was detected in all cultures. Xylanase and pectinase were optimally active at pH 4.5-5.5, while protease was active in a large range of pH 3.5 to 11.0. All three enzymes were maximally active at 40ºC and they were stable for several hours at temperature up to 50ºC.A capacidade do fungo fitopatogênico Myrothecium verrucaria produzir enzimas hidrolíticas extracelulares em culturas submersas foi estudada utilizando diversos substratos. O fungo foi capaz de produzir diferentes depolimerases e glicosidases, sendo xilanases, pectinases e proteases as mais importantes. Atividade lipase foi encontrada nos filtrados das culturas desenvolvidas na presença de óleo de oliva, enquanto atividade proteolítica foi detectada em todas as culturas. Xilanase e pectinase foram otimamente ativas em pH 4,5 a 5,5, enquanto protease foi ativa em ampla faixa de pH (3,5 a 11,0. As três enzimas foram otimamente ativas 40ºC e estáveis por várias horas a temperaturas até 50ºC.

  2. 11 CFR 9032.11 - State.

    Science.gov (United States)

    2010-01-01

    ... 11 Federal Elections 1 2010-01-01 2010-01-01 false State. 9032.11 Section 9032.11 Federal Elections FEDERAL ELECTION COMMISSION PRESIDENTIAL ELECTION CAMPAIGN FUND: PRESIDENTIAL PRIMARY MATCHING FUND DEFINITIONS § 9032.11 State. State means each State of the United States, Puerto Rico, American...

  3. 11 CFR 9002.11 - Qualified campaign expense.

    Science.gov (United States)

    2010-01-01

    ... 11 Federal Elections 1 2010-01-01 2010-01-01 false Qualified campaign expense. 9002.11 Section 9002.11 Federal Elections FEDERAL ELECTION COMMISSION PRESIDENTIAL ELECTION CAMPAIGN FUND: GENERAL ELECTION FINANCING DEFINITIONS § 9002.11 Qualified campaign expense. (a) Qualified campaign expense means...

  4. A comparative study on GM (1,1) and FRMGM (1,1) model in forecasting FBM KLCI

    Science.gov (United States)

    Ying, Sah Pei; Zakaria, Syerrina; Mutalib, Sharifah Sakinah Syed Abd

    2017-11-01

    FTSE Bursa Malaysia Kuala Lumpur Composite Index (FBM KLCI) is a group of indexes combined in a standardized way and is used to measure the Malaysia overall market across the time. Although composite index can give ideas about stock market to investors, it is hard to predict accurately because it is volatile and it is necessary to identify a best model to forecast FBM KLCI. The objective of this study is to determine the most accurate forecasting model between GM (1,1) model and Fourier Residual Modification GM (1,1) (FRMGM (1,1)) model to forecast FBM KLCI. In this study, the actual daily closing data of FBM KLCI was collected from January 1, 2016 to March 15, 2016. GM (1,1) model and FRMGM (1,1) model were used to build the grey model and to test forecasting power of both models. Mean Absolute Percentage Error (MAPE) was used as a measure to determine the best model. Forecasted value by FRMGM (1,1) model do not differ much than the actual value compare to GM (1,1) model for in-sample and out-sample data. Results from MAPE also show that FRMGM (1,1) model is lower than GM (1,1) model for in-sample and out-sample data. These results shown that FRMGM (1,1) model is better than GM (1,1) model to forecast FBM KLCI.

  5. Bioprocessing of wheat bran for the production of lignocellulolytic enzyme cocktail by Cotylidia pannosa under submerged conditions

    Science.gov (United States)

    Sharma, Deepika; Garlapat, Vijay Kumar; Goel, Gunjan

    2016-01-01

    ABSTRACT Characterization and production of efficient lignocellulytic enzyme cocktails for biomass conversion is the need for biofuel industry. The present investigation reports the modeling and optimization studies of lignocellulolytic enzyme cocktail production by Cotylidia pannosa under submerged conditions. The predominant enzyme activities of cellulase, xylanase and laccase were produced in the cocktail through submerged conditions using wheat bran as a substrate. A central composite design approach was utilized to model the production process using temperature, pH, incubation time and agitation as input variables with the goal of optimizing the output variables namely cellulase, xylanase and laccase activities. The effect of individual, square and interaction terms on cellulase, xylanase and laccase activities were depicted through the non-linear regression equations with significant R2 and P-values. An optimized value of 20 U/ml, 17 U/ml and 13 U/ml of cellulase, xylanase and laccase activities, respectively, were obtained with a media pH of 5.0 in 77 h at 31C, 140 rpm using wheatbran as a substrate. Overall, the present study introduces a fungal strain, capable of producing lignocellulolytic enzyme cocktail for subsequent applications in biofuel industry. PMID:26941214

  6. Recombinant human interleukin-11 (IL-11) is a protective factor in severe sepsis with thrombocytopenia: A case-control study.

    Science.gov (United States)

    Wan, Bing; Zhang, Hao; Fu, Haiyan; Chen, Yikun; Yang, Liping; Yin, Jiangtao; Wan, Yin; Shi, Yongqing

    2015-12-01

    To examine the platelet recovering and anti-inflammatory effects of IL-11 in the treatment of sepsis, accompanied with thrombocytopenia and to investigate the associated mechanisms via a case-control study. 105 patients enrolled for the study were segregated into (1) IL-11 therapy group and (2) conventional therapy group. The IL-11 therapy group was given additional recombinant human IL-11 treatment. Laboratory examinations of IL-11, IL-6, TNF-α, PT, APTT, WBC, PLT counts in blood routine assays and PCT, CRP and APACHE II scores were performed and the results were recorded. The PLT counts in the IL-11 therapy group were higher than those in the conventional therapy group. No obvious difference in WBC counts or CRP levels was observed between the two groups. The highest levels of TNF-α were observed on day 3 in the conventional therapy group while it was observed on day 1 in the IL-11 therapy group, both of which subsequently declined gradually. The level of IL-6 was significantly lower in the IL-11 therapy group from 3 to 14 days, while there was a gradual elevation of IL-11. IL-11 therapy downregulated the expression of the sepsis indicator PCT and reduced the APACHE II score from 3 to 14 days. The conventional therapy group had a significantly higher mortality rate within 28 days. IL-11 has a protective role and can accelerate recovery of platelets, and remarkably lessen the extent of inflammatory responses, hence reducing the mortality in sepsis patients accompanied with thrombocytopenia. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Biological nitrate removal from synthetic wastewater using a fungal ...

    African Journals Online (AJOL)

    2007-04-02

    -stage ... grass, instead of non-autoclaved grass containing natural microbial consortia, as sup- ... 25ºC within a 20 ℓ plastic bag, along with a 1 ℓ conical flask ... of cellulases and xylanases – the two major hydrolytic enzyme.

  8. Sequence homolog-based molecular engineering for shifting the enzymatic pH optimum

    Directory of Open Access Journals (Sweden)

    Fuqiang Ma

    2016-09-01

    Full Text Available Cell-free synthetic biology system organizes multiple enzymes (parts from different sources to implement unnatural catalytic functions. Highly adaption between the catalytic parts is crucial for building up efficient artificial biosynthetic systems. Protein engineering is a powerful technology to tailor various enzymatic properties including catalytic efficiency, substrate specificity, temperature adaptation and even achieve new catalytic functions. However, altering enzymatic pH optimum still remains a challenging task. In this study, we proposed a novel sequence homolog-based protein engineering strategy for shifting the enzymatic pH optimum based on statistical analyses of sequence-function relationship data of enzyme family. By two statistical procedures, artificial neural networks (ANNs and least absolute shrinkage and selection operator (Lasso, five amino acids in GH11 xylanase family were identified to be related to the evolution of enzymatic pH optimum. Site-directed mutagenesis of a thermophilic xylanase from Caldicellulosiruptor bescii revealed that four out of five mutations could alter the enzymatic pH optima toward acidic condition without compromising the catalytic activity and thermostability. Combination of the positive mutants resulted in the best mutant M31 that decreased its pH optimum for 1.5 units and showed increased catalytic activity at pH < 5.0 compared to the wild-type enzyme. Structure analysis revealed that all the mutations are distant from the active center, which may be difficult to be identified by conventional rational design strategy. Interestingly, the four mutation sites are clustered at a certain region of the enzyme, suggesting a potential “hot zone” for regulating the pH optima of xylanases. This study provides an efficient method of modulating enzymatic pH optima based on statistical sequence analyses, which can facilitate the design and optimization of suitable catalytic parts for the construction

  9. Caffeine-11C, ephedrine-11C and methylephedrine-11C: synthesis and distribution in mice

    International Nuclear Information System (INIS)

    Saji, Hideo; Ido, Tatsuo; Iwata, Ren; Suzuki, Kazutoshi; Tamate, Kazuhiko

    1978-01-01

    Caffeine, ephedrine and methylephedrine were labeled with carbon-11 by the action of methyliodide- 11 C on theophylline, norephedrine and ephedrine, respectively. Caffeine- 11 C was prepared in 44 min. with a radiochemical yield of 40%, ephedrine- 11 C in 45 min. with a 11% radiochemical yield and methylephedrine- 11 C in 36 min. with a 43% radiochemical yield. When injected in mice intravenously, these products show a high uptake in the liver, the kidney and the blood for caffeine- 11 C and in the liver and the kidney for ephedrine- 11 C and methylephedrine- 11 C. The brain uptake for these products was found to be 2.4 to 3.9% of the injected dose per gram at 5 min. after injection. These studies in mice have demonstrated that these products are potentially useful agents for the dynamic studies of the brain. (auth.)

  10. [11C]copper(I) cyanide: a new radioactive precursor or 11C-cyanation and functionalization of haloarenes

    International Nuclear Information System (INIS)

    Ponchant, M.; Hinnen, F.; Demphel, S.; Crouzel, C.

    1997-01-01

    Radiosyntheses of [cyano- 11 C]benzonitriles, [carboyxyl- 11 C]benzoic acids, [carbonyl- 11 C]benzamides and [tetrazoyl- 11 C]5-biphenyl-1H-tetrazoles are described. [Cyano- 11 C]benzonitriles were prepared using the Rosenmund-von Braun reaction in one step from [ 11 C]hydrogen cyanide via [ 11 C]copper(1) cyanide in 2 min. [Carboxyl- 11 C]benzoic acids and [carbonyl- 11 C]benzamides were prepared from [cyano- 11 C]benzonitrile-complexes using basic hydrogen peroxide for 3 min in N,N-dimethylformamide at 180 and 80 o C, respectively. [Tetrazoyl- 11 C]5-biphenyl-1H-tetrazoles were obtained from [cyano- 11 C]benzonitrile-complexes in 7 min by cyclization with trimethyltin azide followed by acid hydrolysis at 180 o C. These labelled compounds were obtained in a one-pot procedure with good average specific radioactivities [500 mCi/μmol) at the end of synthesis] and in satisfactory average radiochemical yields (24-69%) at 40 min from the end of radionuclide production. This new radioactive precursor, [ 11 C]copper(I) cyanide, should give access to new 11 C-labelled ligands for positron emission tomography. (author)

  11. Effects of enzymes to improve sensory quality of frozen dough bread and analysis on its mechanism.

    Science.gov (United States)

    Wang, Xuan; Pei, Dudu; Teng, Yuefei; Liang, Jianfen

    2018-01-01

    Baking quality of frozen dough is negatively affected by dough weakening and by a reduction in both yeast viability and activity during freezing and frozen storage. The objective of this study was to investigate effects of different enzymes, such as α-amylase, xylanase, celluase, glucose oxidase, and lipase on the texture and sensory quality of bread after frozen storage, as well as on dough properties, in terms of fermentation characteristics, freezable water contents and microstructure. Except for α-amylase, other enzymes improved the bread sensory quality and got higher overall acceptability, especially xylanase. Dough fermentative behavior showed that the maximum heights of frozen dough were increased by 33.2, 19.7 and 7.4%, respectively with xylanase, cellulase and lipase. Cellulase lowered gas holding ability of dough. Thermodynamic properties indicated that addition of enzyme decreased the freezable water contents in frozen dough. Scanning electronic microscopy revealed that freezing and frozen storage disrupted dough gluten network causing separation of starch granules from the gluten matrix. Inclusion of cellulase, xylanase and lipase made the frozen dough having a more continuous gluten network and smoother surface, and glucose oxidase increased the stability of the gluten work.

  12. Monosaccharides and Ethanol Production from Superfine Ground Sugarcane Bagasse Using Enzyme Cocktail

    Directory of Open Access Journals (Sweden)

    Jingbo Li

    2014-03-01

    Full Text Available In this work, the effect of particle size on the enzymatic hydrolysis of milled and sieved sugarcane bagasse (SCB was studied. The enzymatic hydrolysis and fermentability of superfine ground SCB (SGP400 using an enzyme cocktail strategy were also explored. Particle size reduction improved the enzymatic hydrolysis. The highest glucose yield was 44.75%, which was obtained from SGP400. The enzyme cocktail strategy greatly enhanced the glucose and xylose yield. The maximum glucose and xylose yield was from the enzyme cocktail of cellulase, xylanase, and pectinase. Synergistic action between xylanase and pectinase as well as cellulase and pectinase was quite noticeable. Hydrolysis times affected the degree of synergism. Ethanol production was carried out by employing simultaneous saccharification and fermentation (SSF and semi-SSF using enzymes and their cocktails. Semi-SSF was found to be the better one compared with SSF. Xylanase and pectinase aided the ethanol production in both fermentation modes. Ethanol yield was 7.81 and 7.30 g/L for semi-SSF and SSF, respectively by using an enzyme cocktail of cellulase, β-glucosidase, pectinase, and xylanase.

  13. Effects of storage temperature on survival and infectivity of three indigenous entomopathogenic nematodes strains (Steinernematidae and Heterorhabditidae) from Meghalaya, India.

    Science.gov (United States)

    Lalramliana; Yadav, Arun K

    2016-12-01

    Three locally isolated strains of entomopathogenic nematodes (EPNs), viz. Heterorhabditis indica , Steinernema thermophilum and Steinernema glaseri , from Meghalaya, India were characterized in terms of storage temperature and survival and infectivity of their infective juveniles (IJs). The survival and infectivity of nematode IJs was studied at, 5 ± 2 and 25 ± 2 °C, for a period of 120 days, using deionized water as storage medium. The viability of nematode IJs was checked by mobility criterion at different storage periods, while the infectivity of nematode IJs was ascertained on the basis of establishment of IJs, using Galleria mellonella larva mortality tests in petridishes. The results of this study revealed that storage temperature markedly affects the survival as well as the establishment of nematode IJs of the three EPN species. At 5 °C, comparatively higher rate of IJ's survival (i.e. 74-86 %) was observed for 15 days of storage, but the same reduced drastically to 28-32 % after 30 days of storage for H. indica and S. thermophilum . On the other hand, at 25 °C, the survival of nematode IJs was observed till 120 days for all the three studied EPNs. In case of S. thermophilum and S. glaseri , higher rate of IJs survival (>75 %) was observed respectively at 15 and 30 days of observation. The study also showed that the establishment of IJs of the three EPN species declines with increase in storage periods, at both the test temperatures. In general, the nematodes stored at 25 °C showed comparatively better establishment than those stored at 5 °C. Among the three EPN studied, the establishment of S. glaseri was comparatively better than the rest of the species at both the temperatures and for different storage durations. In conclusion, our study adds further valuable information about the effect of storage temperature on survival and infectivity of three indigenous EPN species of Meghalaya, India which appears to be promising biocontrol

  14. Enzymatic production of wheat and ryegrass derived xylooligosaccharides and evaluation of their in vitro effect on pig gut microbiota

    DEFF Research Database (Denmark)

    Dotsenko, Gleb; Meyer, Anne S.; Canibe, Nuria

    2017-01-01

    This study examines enzymatic production of linear xylooligosaccharides (XOS) and branched arabinoxylooligosaccharides (AXOS) from monocotyledonous biomass, wheat straw and ryegrass, and compares the in vitro effects of these XOS and AXOS on pig gut microbiota. XOS and AXOS were obtained from...... the biomass by treatment with different endo-1,4-β-xylanases. XOS of DP2-6 from wheat straw, obtained after treatment with Aspergillus niger endo GH11, suppressed growth of Clostridium perfringens and resulted in a high level of lactic acid production when fermented in vitro by pig fecal microbiota...... (GH11). These results indicate that wheat straw as well as green grass biomass such as ryegrass have potential as new sources of putative prebiotics for pig feed....

  15. Consistent mutational paths predict eukaryotic thermostability

    Directory of Open Access Journals (Sweden)

    van Noort Vera

    2013-01-01

    Full Text Available Abstract Background Proteomes of thermophilic prokaryotes have been instrumental in structural biology and successfully exploited in biotechnology, however many proteins required for eukaryotic cell function are absent from bacteria or archaea. With Chaetomium thermophilum, Thielavia terrestris and Thielavia heterothallica three genome sequences of thermophilic eukaryotes have been published. Results Studying the genomes and proteomes of these thermophilic fungi, we found common strategies of thermal adaptation across the different kingdoms of Life, including amino acid biases and a reduced genome size. A phylogenetics-guided comparison of thermophilic proteomes with those of other, mesophilic Sordariomycetes revealed consistent amino acid substitutions associated to thermophily that were also present in an independent lineage of thermophilic fungi. The most consistent pattern is the substitution of lysine by arginine, which we could find in almost all lineages but has not been extensively used in protein stability engineering. By exploiting mutational paths towards the thermophiles, we could predict particular amino acid residues in individual proteins that contribute to thermostability and validated some of them experimentally. By determining the three-dimensional structure of an exemplar protein from C. thermophilum (Arx1, we could also characterise the molecular consequences of some of these mutations. Conclusions The comparative analysis of these three genomes not only enhances our understanding of the evolution of thermophily, but also provides new ways to engineer protein stability.

  16. Aquaporin-11 (AQP11 Expression in the Mouse Brain

    Directory of Open Access Journals (Sweden)

    Shin Koike

    2016-06-01

    Full Text Available Aquaporin-11 (AQP11 is an intracellular aquaporin expressed in various tissues, including brain tissues in mammals. While AQP11-deficient mice have developed fatal polycystic kidneys at one month old, the role of AQP11 in the brain was not well appreciated. In this study, we examined the AQP11 expression in the mouse brain and the brain phenotype of AQP11-deficient mice. AQP11 messenger ribonucleic acid (mRNA and protein were expressed in the brain, but much less than in the thymus and kidney. Immunostaining showed that AQP11 was localized at the epithelium of the choroid plexus and at the endothelium of the brain capillary, suggesting that AQP11 may be involved in water transport at the choroid plexus and blood-brain barrier (BBB in the brain. The expression of AQP4, another brain AQP expressed at the BBB, was decreased by half in AQP11-deficient mice, thereby suggesting the presence of the interaction between AQP11 and AQP4. The brain of AQP11-deficient mice, however, did not show any morphological abnormalities and the function of the BBB was intact. Our findings provide a novel insight into a water transport mechanism mediated by AQPs in the brain, which may lead to a new therapy for brain edema.

  17. Synthesis and preclinical evaluation of [11C]D617, a metabolite of (R)-[11C]verapamil

    NARCIS (Netherlands)

    Verbeek, Joost; Syvänen, Stina; Schuit, Robert C.; Eriksson, Jonas; de Lange, Elizabeth C M; Windhorst, Albert D.; Luurtsema, Gert; Lammertsma, Adriaan A.

    2012-01-01

    OBJECTIVES: (R)-[(11)C]verapamil is widely used as a positron emission tomography (PET) tracer to evaluate P-glycoprotein (P-gp) functionality at the blood-brain barrier in man. A disadvantage of (R)-[(11)C]verapamil is the fact that its main metabolite, [(11)C]D617, also enters the brain. For

  18. A molecular analysis of (hemi-)cellulose degradation by Aspergilli

    NARCIS (Netherlands)

    Gielkens, M.M.C.

    1999-01-01

    Glycosylhydrolases like cellulases and xylanases are of great importance for the ecological recycling of biomass. The saprophytic fungi, e.g Aspergillus niger , are capable of degrading plant cell wall material by secreting these enzymes. Because of their properties, a

  19. Simulasi Kinerja Jaringan Nirkabel IEEE-802.11a dan IEEE-802.11g Menggunakan NS-2

    Directory of Open Access Journals (Sweden)

    Helm Fitriawan

    2014-03-01

    Full Text Available Wireless network uses transmission media based on radio waves. This type of networks is mainly useddue to its efficiency and mobility in data exchanging. This paper reports the modeling and simulation of wirelessnetworks based on Cisco Aironet 1130ag access point devices with IEEE 802.11a and IEEE 802.11g standards. Themodeling and simulation are performed using network simulator version 2 (NS-2 that is installed on operationsystem Linux Ubuntu v.10.10. The NS-2 is commonly used and works well in numerous types of network simulation. From simulation, we obtain quality of service parameters by employing several simulation scenarios in terms ofnumber of nodes, distances, and packet data sizes. It can be concluded from simulation results that the IEEE 802.11gnetworks transfer data with better quality than those of IEEE 802.11a networks.  Furthermore, the IEEE 802.11gnetworks provide a higher throughput, with smaller amount of delay and packet loss percentage compared to thoseof IEEE 802.11a networks.

  20. Comparative evaluation of fermented and non-fermented de-oiled rice bran with or without exogenous enzymes supplementation in the diet of Labeo rohita (Hamilton, 1822).

    Science.gov (United States)

    Ranjan, Amit; Sahu, Narottam Prasad; Deo, Ashutosh Dharmendra; Kumar, H Sanath; Kumar, Sarvendra; Jain, Kamal Kant

    2018-03-29

    A 60-day feeding trial was conducted to study the effect of exogenous enzymes (xylanase and phytase) supplementation in the non-fermented and fermented de-oiled rice bran (DORB)-based diet of Labeo rohita. Four test diets (T1-DORB-based diet, T2-fermented DORB-based diet, T3-phytase and xylanase supplemented DORB-based diet, and T4-phytase and xylanase supplemented fermented DORB-based diet) were formulated and fed to the respective groups. Test diets T3 and T4 were supplemented with 0.01% xylanase (16,000 U kg -1 ) and 0.01% phytase (500 U kg -1 ) enzymes. One hundred twenty juveniles of L. rohita, with an average weight 5.01 ± 0.02 g, were stocked in 12 uniform size plastic rectangular tanks in triplicate with 10 fishes per tank following a completely randomized design (CRD). Exogenous enzyme supplementation to the T3 group significantly improved the growth performance of L. rohita (p exogenous enzymes. The carcass composition (except CP %), enzyme activities (except amylase activity), globulin, and A/G ratio did not vary significantly (p > 0.05). Based on the results of the present study, it is concluded that exogenous enzyme supplementation significantly increases the growth of fish fed with DORB-based diet.

  1. 37 CFR 11.11 - Administrative suspension, inactivation, resignation, and readmission.

    Science.gov (United States)

    2010-07-01

    ... 37 Patents, Trademarks, and Copyrights 1 2010-07-01 2010-07-01 false Administrative suspension, inactivation, resignation, and readmission. 11.11 Section 11.11 Patents, Trademarks, and Copyrights UNITED... Other Non-Patent Law § 11.11 Administrative suspension, inactivation, resignation, and readmission. (a...

  2. Tracers development for the PET study of nicotinic receptors: [11C]-mecamylamine and [11C]-SIB 1553A. Tritium and carbon-11 radiolabelling of a serine proteinase inhibitor: the t-PAstop

    International Nuclear Information System (INIS)

    Sobrio, F.

    2002-12-01

    In order to develop radiotracers for the Positron Emission Tomography (PET), we labelled both the mecamylamine and SIB-1553A with carbon-11 to study the nicotinic cholinergic receptors (nAChRs). The radiosynthesis of [ 11 C]-t-PA stop and the labelling with tritium of one analogue were realized for cerebral ischemia PET studies. The [ 11 C]-mecamylamine, a non-competitive and non-selective nAChRs antagonist was synthesized in 45 min via a N-[ 11 C]-methylation reaction. In the rat brain, the ex vivo studies showed no radio-metabolite 45 min after the injection of [ 11 C]-mecamylamine. The uptake kinetics in the rat brain or in vivo by PET in the anesthetized baboon or in the conscious monkey, reached a plateau around 45-50 min after injection. However, the saturation or displacement experiments did not permit to exhibit nor a significant difference of labelling between the different cerebral regions nor a specific uptake. In consequence, the [ 11 C]-mecamylamine was not an appropriate radioligand for nAChRs PET study. The labelling of [ 11 C]-SIB 1553A, a selective agonist for the nicotinic β4 subunit, required the synthesis in 5 steps (56% overall yield) of precursor for the incorporation of carbon-11. The radiosynthesis was performed in 36 min by a N-[ 11 C]-methylation reaction (yield: 75%). The [ 11 C]-t-PA stop was obtained from [ 11 C]-KCN with yields from 80 to 90%. For the first time with carbon-11, the formation of an amidine group was realized from a nitrile group. The labelling by isotopic exchange of hydrogen by tritium of the t-PA stop did not permit to obtain the [ 3 H]-t-PA stop but a tritiated analogue. This compound will be used to study its vectorization by micro-encapsulation. (author)

  3. OsPEX11, a peroxisomal biogenesis factor 11, contributes to salt stress tolerance in Oryza sativa

    Directory of Open Access Journals (Sweden)

    Cui Peng

    2016-09-01

    Full Text Available Peroxisomes are single membrane-bound organelles, whose basic enzymatic constituents are catalase and H2O2-producing flavin oxidases. Previous reports showed that peroxisome is involved in numerous processes including primary and secondary metabolism, plant development and abiotic stress responses. However, knowledge on the function of different peroxisome genes from rice and its regulatory roles in salt and other abiotic stresses is limited. Here, a novel prey protein, OsPEX11 (Os03g0302000, was screened and identified by yeast two-hybrid and GST pull down assays. Phenotypic analysis of OsPEX11 overexpression seedlings demonstrated that they had better tolerance to salt stress than wild type and OsPEX11-RNAi seedlings. Compared with wild type and OsPEX11-RNAi seedlings, overexpression of OsPEX11 had lower level of lipid peroxidation, Na+/K+ ratio, higher activities of antioxidant enzymes (SOD, POD and CAT and proline accumulation. Furthermore, qPCR data suggested that OsPEX11 acted as a positive regulator of salt tolerance by reinforcing the expression of several well-known rice transporters (OsHKT2;1, OsHKT1;5, OsLti6a, OsLti6b, OsSOS1, OsNHX1 and OsAKT1 involved in Na+/K+ homeostasis in transgenic plants under salinity. Ultrastructural observations of OsPEX11-RNAi seedlings showed that they were less sensitive to salt stress than wild type and overexpression lines. These results provide experimental evidence that OsPEX11 is an important gene implicated in Na+ and K+ regulation, and plays a critical role in salt stress tolerance by modulating the expression of cation transporters and antioxidant defense. Thus, OsPEX11 could be considered in transgenic breeding for improvement of salt stress tolerance in rice crop.

  4. OsPEX11, a Peroxisomal Biogenesis Factor 11, Contributes to Salt Stress Tolerance in Oryza sativa.

    Science.gov (United States)

    Cui, Peng; Liu, Hongbo; Islam, Faisal; Li, Lan; Farooq, Muhammad A; Ruan, Songlin; Zhou, Weijun

    2016-01-01

    Peroxisomes are single membrane-bound organelles, whose basic enzymatic constituents are catalase and H 2 O 2 -producing flavin oxidases. Previous reports showed that peroxisome is involved in numerous processes including primary and secondary metabolism, plant development and abiotic stress responses. However, knowledge on the function of different peroxisome genes from rice and its regulatory roles in salt and other abiotic stresses is limited. Here, a novel prey protein, OsPEX11 (Os03g0302000), was screened and identified by yeast two-hybrid and GST pull-down assays. Phenotypic analysis of OsPEX11 overexpression seedlings demonstrated that they had better tolerance to salt stress than wild type (WT) and OsPEX11-RNAi seedlings. Compared with WT and OsPEX11-RNAi seedlings, overexpression of OsPEX11 had lower level of lipid peroxidation, Na + /K + ratio, higher activities of antioxidant enzymes (SOD, POD, and CAT) and proline accumulation. Furthermore, qPCR data suggested that OsPEX11 acted as a positive regulator of salt tolerance by reinforcing the expression of several well-known rice transporters ( OsHKT2;1, OsHKT1;5, OsLti6a, OsLti6b, OsSOS1, OsNHX1 , and OsAKT1 ) involved in Na + /K + homeostasis in transgenic plants under salinity. Ultrastructural observations of OsPEX11-RNAi seedlings showed that they were less sensitive to salt stress than WT and overexpression lines. These results provide experimental evidence that OsPEX11 is an important gene implicated in Na + and K + regulation, and plays a critical role in salt stress tolerance by modulating the expression of cation transporters and antioxidant defense. Thus, OsPEX11 could be considered in transgenic breeding for improvement of salt stress tolerance in rice crop.

  5. Synthesis of n. c. a. PET-radiotracers with carbon-11. Zur Synthese traegerarme PET-Radiotracer mit Kohlenstoff-11

    Energy Technology Data Exchange (ETDEWEB)

    Schirbel, A.

    1998-11-01

    Carbon-11 offers the unique possibility of authentic labelling of molecules as radioindicators for non invasive and quantitative determination of physiological functions via positron emission tomography (PET). Therefore, the goal of this thesis was to synthesize of different n.c.a. [sup 11]C-labelled pharmaceuticals for in vivo distribution studies with PET. For the determination of the pharmacokinetics of [1-[sup 11]C]acetate in porcine myocardium during prolonged ischemia, n.c.a. [1-[sup 11]C]acetate was synthesized via carboxylation of methylmagnesium bromide with in target produced n.c.a. [[sup 11]C]CO[sub 2] with a radiochemical yield (RCY) of 68 [+-] 7%. The fast (18 min) and reliable radiosynthesis allowed for repeated tracer administration at short intervals (<20 min). In order to study the pharmacokinetics and metabolism of acetylsalicylic acid (Aspirin[sup R]c) in humans, [1-[sup 11]C]acetylsalicylic acid, acetyl-[carboxy-[sup 11]C]salicylic acid and [carboxy-[sup 11]C]salicylic acid were prepared. N.c.a. [1-[sup 11]C]acetylsalicylic acid was synthesized via the reaction of [1-[sup 11]C]acetylchloride with salicylic acid salts. The use of the silver salt proved to be superior to the sodium salt and resulted in radiochemical yields of 32 [+-] 5%. Base-line (clean) separation of the labelled product was achieved using radio-HPLC. With regard to the preparation of n.c.a. [carboxy-[sup 11]C]salicylic acid, several protected and unprotected phenol derivates were metallated and subsequently carboxylated using n.c.a. [[sup 11]C]CO[sub 2]. Best results (87 [+-] 3% RCY) could be achieved with 2-(methoxymethoxy)-phenylmagnesium iodide as a precursor and subsequent quantitative cleavage of the MOM-group. Acetylation of n.c.a. [carboxy-[sup 11]C]salicylic acid to acetyl-[carboxy-[sup 11]C]salicylic acid was performed using acetylchloride in CH[sub 2]Cl[sub 2] with a radiochemical yield of 65 [+-] 4%. (orig.)

  6. Effects of pyrite and sphalerite on population compositions, dynamics and copper extraction efficiency in chalcopyrite bioleaching process.

    Science.gov (United States)

    Xiao, Yunhua; Liu, Xueduan; Dong, Weiling; Liang, Yili; Niu, Jiaojiao; Gu, Yabing; Ma, Liyuan; Hao, Xiaodong; Zhang, Xian; Xu, Zhen; Yin, Huaqun

    2017-07-01

    This study used an artificial microbial community with four known moderately thermophilic acidophiles (three bacteria including Acidithiobacillus caldus S1, Sulfobacillus thermosulfidooxidans ST and Leptospirillum ferriphilum YSK, and one archaea, Ferroplasma thermophilum L1) to explore the variation of microbial community structure, composition, dynamics and function (e.g., copper extraction efficiency) in chalcopyrite bioleaching (C) systems with additions of pyrite (CP) or sphalerite (CS). The community compositions and dynamics in the solution and on the ore surface were investigated by real-time quantitative PCR (qPCR). The results showed that the addition of pyrite or sphalerite changed the microbial community composition and dynamics dramatically during the chalcopyrite bioleaching process. For example, A. caldus (above 60%) was the dominant species at the initial stage in three groups, and at the middle stage, still dominated C group (above 70%), but it was replaced by L. ferriphilum (above 60%) in CP and CS groups; at the final stage, L. ferriphilum dominated C group, while F. thermophilum dominated CP group on the ore surface. Furthermore, the additions of pyrite or sphalerite both made the increase of redox potential (ORP) and the concentrations of Fe 3+ and H + , which would affect the microbial community compositions and copper extraction efficiency. Additionally, pyrite could enhance copper extraction efficiency (e.g., improving around 13.2% on day 6) during chalcopyrite bioleaching; on the contrary, sphalerite restrained it.

  7. Macro-IML manual for DEC PDP 11 computer with controller DEC CA 11-A/BORER type 1,533 A

    International Nuclear Information System (INIS)

    Kubitz, M.; Kind, R.

    1975-03-01

    The IML-implementations follow the Macro-Syntax as given in Appendix A of the document 'CAMAC. The Definitior of IML (A Language For Use in CAMAC Systems)'. This document has been adopted as a description by ESONE and AEC NIM in August/September 74 and has been published in October 74. They have been designed for the DEC PDP 11 computer with the branch controller DEC CA 11-A and the Single Crate Controller BORER Type 1,533 A. For both DEC operating systems, DOS V08/09 and RSX-11D/M a full set of macros has been implemented except the block transfer on special LAM, X-error control statements and the subscript mode. Transfer modes not implemented by the hardware of the CA 11-A are simulated by software. (orig.) [de

  8. Frequent phosphodiesterase 11A gene (PDE11A) defects in patients with Carney complex (CNC) caused by PRKAR1A mutations: PDE11A may contribute to adrenal and testicular tumors in CNC as a modifier of the phenotype.

    Science.gov (United States)

    Libé, Rossella; Horvath, Anelia; Vezzosi, Delphine; Fratticci, Amato; Coste, Joel; Perlemoine, Karine; Ragazzon, Bruno; Guillaud-Bataille, Marine; Groussin, Lionel; Clauser, Eric; Raffin-Sanson, Marie-Laure; Siegel, Jennifer; Moran, Jason; Drori-Herishanu, Limor; Faucz, Fabio Rueda; Lodish, Maya; Nesterova, Maria; Bertagna, Xavier; Bertherat, Jerome; Stratakis, Constantine A

    2011-01-01

    Carney complex (CNC) is an autosomal dominant multiple neoplasia, caused mostly by inactivating mutations of the regulatory subunit 1A of the protein kinase A (PRKAR1A). Primary pigmented nodular adrenocortical disease (PPNAD) is the most frequent endocrine manifestation of CNC with a great inter-individual variability. Germline, protein-truncating mutations of phosphodiesterase type 11A (PDE11A) have been described to predispose to a variety of endocrine tumors, including adrenal and testicular tumors. Our objective was to investigate the role of PDE11A as a possible gene modifier of the phenotype in a series of 150 patients with CNC. A higher frequency of PDE11A variants in patients with CNC compared with healthy controls was found (25.3 vs. 6.8%, P CNC patients, those with PPNAD were significantly more frequently carriers of PDE11A variants compared with patients without PPNAD (30.8 vs. 13%, P = 0.025). Furthermore, men with PPNAD were significantly more frequently carriers of PDE11A sequence variants (40.7%) than women with PPNAD (27.3%) (P CNC patients, a high frequency of PDE11A variants, suggesting that PDE11A is a genetic modifying factor for the development of testicular and adrenal tumors in patients with germline PRKAR1A mutation.

  9. Protein crystallography with a micrometre-sized synchrotron-radiation beam

    International Nuclear Information System (INIS)

    Moukhametzianov, Rouslan; Burghammer, Manfred; Edwards, Patricia C.; Petitdemange, Sebastien; Popov, Dimitri; Fransen, Maikel; McMullan, Gregory; Schertler, Gebhard F. X.; Riekel, Christian

    2008-01-01

    For the first time, protein microcrystallography has been performed with a focused synchrotron-radiation beam of 1 µm using a goniometer with a sub-micrometre sphere of confusion. The crystal structure of xylanase II has been determined with a flux density of about 3 × 10 10 photons s −1 µm −2 at the sample. For the first time, protein microcrystallography has been performed with a focused synchrotron-radiation beam of 1 µm using a goniometer with a sub-micrometre sphere of confusion. The crystal structure of xylanase II has been determined with a flux density of about 3 × 10 10 photons s −1 µm −2 at the sample. Two sets of diffraction images collected from different sized crystals were shown to comprise data of good quality, which allowed a 1.5 Å resolution xylanase II structure to be obtained. The main conclusion of this experiment is that a high-resolution diffraction pattern can be obtained from 20 µm 3 crystal volume, corresponding to about 2 × 10 8 unit cells. Despite the high irradiation dose in this case, it was possible to obtain an excellent high-resolution map and it could be concluded from the individual atomic B-factor patterns that there was no evidence of significant radiation damage. The photoelectron escape from a narrow diffraction channel is a possible reason for reduced radiation damage as indicated by Monte Carlo simulations. These results open many new opportunities in scanning protein microcrystallography and make random data collection from microcrystals a real possibility, therefore enabling structures to be solved from much smaller crystals than previously anticipated as long as the crystallites are well ordered

  10. Evaluation of (+)-p-[11C]methylvesamicol for mapping sigma1 receptors: a comparison with [11C]SA4503

    International Nuclear Information System (INIS)

    Ishiwata, Kiichi; Kawamura, Kazunori; Yajima, Kazuyoshi; QingGeLeTu; Mori, Hirofumi; Shiba, Kazuhiro

    2006-01-01

    Vesamicol is a leading compound for positron emission tomography (PET) and single photon emission computed tomography (SPECT) tracers for mapping the vesicular acetylcholine transporter (VAChT). Recently, we found that (+)-p-methylvesamicol ((+)-PMV) has low affinity for VAChT (K i =199 nM), but has moderate to high affinity for sigma receptors: K i =3.0 nM for sigma 1 and K i =40.7 nM for sigma 2 , and that sigma 1 -selective SA4503 (K i =4.4 nM for sigma 1 and K i =242 nM for sigma 2 ) has moderate affinity for VAChT (K i =50.2 nM). In the present study, we examined the potential of (+)-[ 11 C]PMV as a PET radioligand for mapping sigma 1 receptors as compared with [ 11 C]SA4503. In rat brain, similar regional distribution patterns of (+)-[ 11 C]PMV and [ 11 C]SA4503 were shown by tissue dissection and by ex vivo autoradiography. Blocking experiments using (±)-PMV (-)-vesamicol, SA4503, haloperidol and (±)-pentazocine showed that the two tracers specifically bound to sigma 1 receptors, and that [ 11 C]SA4503 exhibited greater specific binding than (+)-[ 11 C]PMV. No sign of VAChT-specific binding by [ 11 C]SA4503 was observed in the striatum, which is rich in VAChT sites. In conclusion, (+)-[ 11 C]PMV specifically bound to sigma 1 receptors in the brain, but to a lesser extent than [ 11 C]SA4503, suggesting that (+)-[ 11 C]PMV is a less preferable PET ligand than [ 11 C]SA4503. On the other hand, the moderate affinity of [ 11 C]SA4503 for VAChT is negligible in vivo

  11. Extracellular enzyme activities during lignocellulose degradation by Streptomyces spp.: a comparative study of wild-type and genetically manipulated strains

    International Nuclear Information System (INIS)

    Ramachandra, M.; Crawford, D.L.; Pometto, A.L. III

    1987-01-01

    The wild-type ligninolytic actinomycete Streptomyces viridosporus T7A and two genetically manipulated strains with enhanced abilities to produce a water-soluble lignin degradation intermediate, an acid-precipitable polymeric lignin (APPL), were grown on lignocellulose in solid-state fermentation cultures. Culture filtrates were periodically collected, analyzed for APPL, and assayed for extracellular lignocellulose-catabolizing enzyme activities. Two APPL-overproducing strains, UV irradiation mutant T7A-81 and protoplast fusion recombinant SR-10, had higher and longer persisting peroxidase, esterase, and endoglucanase activities than did the wild-type strain T7A. Results implicated one or more of these enzymes in lignin solubilization. Only mutant T7A-81 had higher xylanase activity than the wild type. The peroxidase was induced by both lignocellulose and APPL. This extracellular enzyme has some similarities to previously described ligninases in fungi. This is the first report of such an enzyme in Streptomyces spp. Four peroxidase isozymes were present, and all catalyzed the oxidation of 3,4-dihydroxyphenylalanine, while one also catalyzed hydrogen peroxide-dependent oxidation of homoprotocatechuic acid and caffeic acid. Three constitutive esterase isozymes were produced which differed in substrate specificity toward α-naphthyl acetate and α-naphthyl butyrate. Three endoglucanase bands, which also exhibited a low level of xylanase activity, were identified on polyacrylamide gels as was one xylanase-specific band. There were no major differences in the isoenzymes produced by the different strains. The probable role of each enzyme in lignocellulose degradation is discussed

  12. Investigation of the indigenous fungal community populating barley grains: Secretomes and xylanolytic potential.

    Science.gov (United States)

    Sultan, Abida; Frisvad, Jens C; Andersen, Birgit; Svensson, Birte; Finnie, Christine

    2017-10-03

    The indigenous fungal species populating cereal grains produce numerous plant cell wall-degrading enzymes including xylanases, which could play important role in plant-pathogen interactions and in adaptation of the fungi to varying carbon sources. To gain more insight into the grain surface-associated enzyme activity, members of the populating fungal community were isolated, and their secretomes and xylanolytic activities assessed. Twenty-seven different fungal species were isolated from grains of six barley cultivars over different harvest years and growing sites. The isolated fungi were grown on medium containing barley flour or wheat arabinoxylan as sole carbon source. Their secretomes and xylanase activities were analyzed using SDS-PAGE and enzyme assays and were found to vary according to species and carbon source. Secretomes were dominated by cell wall degrading enzymes with xylanases and xylanolytic enzymes being the most abundant. A 2-DE-based secretome analysis of Aspergillus niger and the less-studied pathogenic fungus Fusarium poae grown on barley flour and wheat arabinoxylan resulted in identification of 82 A. niger and 31 F. poae proteins many of which were hydrolytic enzymes, including xylanases. The microorganisms that inhabit the surface of cereal grains are specialized in production of enzymes such as xylanases, which depolymerize plant cell walls. Integration of gel-based proteomics approach with activity assays is a powerful tool for analysis and characterization of fungal secretomes and xylanolytic activities which can lead to identification of new enzymes with interesting properties, as well as provide insight into plant-fungal interactions, fungal pathogenicity and adaptation. Understanding the fungal response to host niche is of importance to uncover novel targets for potential symbionts, anti-fungal agents and biotechnical applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Identification of Pectin Degrading Enzymes Secreted by Xanthomonas oryzae pv. oryzae and Determination of Their Role in Virulence on Rice

    OpenAIRE

    Tayi, Lavanya; Maku, Roshan V.; Patel, Hitendra Kumar; Sonti, Ramesh V.

    2016-01-01

    Xanthomonas oryzae pv.oryzae (Xoo) causes the serious bacterial blight disease of rice. Xoo secretes a repertoire of plant cell wall degrading enzymes (CWDEs) like cellulases, xylanases, esterases etc., which act on various components of the rice cell wall. The major cellulases and xylanases secreted by Xoo have been identified and their role in virulence has been determined. In this study, we have identified some of the pectin degrading enzymes of Xoo and assessed their role in virulence. Bi...

  14. Carbon 11 labelled phosgene: a new synthesis - medical interest

    International Nuclear Information System (INIS)

    Landais, P.

    1985-01-01

    This thesis describes a new synthesis of high specific radioactivity carbon-11 labelled phosgene. The latter is an important precursor for the labelling of radiopharmaceuticals used in Positron Emission Tomography. The synthesis is carried out in 10 minutes. First, the carbon-11 labelled methane ( 11 CH 4 ) is chlorinated into carbon tetrachloride on pumice impregnated with copper (II) chloride. A photochemical process had previously been studied but this reaction was strongly inhibited. Then the 11 C-carbon tetrachloride is oxidized into 11 C-phosgene on hot stainless. The 11 C-CGP 12177 has been labelled from this new 11 C-Phosgene synthesis for receptor studies which require high specific radioactivity. (author) [fr

  15. Investigation of the indigenous fungal community populating barley grains: Secretomes and xylanolytic potential

    DEFF Research Database (Denmark)

    Sultan, Abida; Frisvad, Jens Christian; Andersen, Birgit

    2017-01-01

    The indigenous fungal species populating cereal grains produce numerous plant cell wall-degrading enzymes including xylanases, which could play important role in plant-pathogen interactions and in adaptation of the fungi to varying carbon sources. To gain more insight into the grain surface......-associated enzyme activity, members of the populating fungal community were isolated, and their secretomes and xylanolytic activities assessed. Twenty-seven different fungal species were isolated from grains of six barley cultivars over different harvest years and growing sites. The isolated fungi were grown...... on medium containing barley flour or wheat arabinoxylan as sole carbon source. Their secretomes and xylanase activities were analyzed using SDS-PAGE and enzyme assays and were found to vary according to species and carbon source. Secretomes were dominated by cell wall degrading enzymes with xylanases...

  16. Biological nitrate removal from synthetic wastewater using a fungal ...

    African Journals Online (AJOL)

    A series of lignocellulosic fungi, capable of cellulase and/or xylanase production, were isolated from soil to be used for cellulose degradation and nitrate removal from nitrate-rich wastewater in simple one-stage anaerobic bioreactors containing grass cuttings as source of cellulose. The fungal consortium, consisting of six ...

  17. Synthesis of O-[11C]acetyl CoA, O-[11C]acetyl-L-carnitine, and L-[11C]carnitine labelled in specific positions, applied in PET studies on rhesus monkey

    International Nuclear Information System (INIS)

    Jacobson, Gunilla B.; Watanabe, Yasuyoshi; Valind, Sven; Kuratsune, Hirohiko; Laangstroem, Bengt

    1997-01-01

    The syntheses of L-carnitine, O-acetyl CoA, and O-acetyl-L-carnitine labelled with 11 C at the 1- or 2-position of the acetyl group or the N-methyl position of carnitine, using the enzymes acetyl CoA synthetase and carnitine acetyltransferase, are described. With a total synthesis time of 45 min, O-[1- 11 C]acetyl CoA and O-[2- 11 C]acetyl CoA was obtained in 60-70% decay-corrected radiochemical yield, and O-[1- 11 C]acetyl-L-carnitine and O-[2- 11 C]acetyl-L-carnitine in 70-80% yield, based on [1- 11 C]acetate or [2- 11 C]acetate, respectively. By an N-methylation reaction with [ 11 C]methyl iodide, L-[methyl- 11 C]carnitine was obtained within 30 min, and O-acetyl-L-[methyl- 11 C]carnitine within 40 min, giving a decay-corrected radiochemical yield of 60% and 40-50%, respectively, based on [ 11 C]methyl iodide. Initial data of the kinetics of the different 11 C-labelled L-carnitine and acetyl-L-carnitines in renal cortex of anaesthetized monkey (Macaca mulatta) are presented

  18. Plant cell-wall hydrolyzing enzymes from indigenously isolated fungi grown on conventional and novel natural substrates

    International Nuclear Information System (INIS)

    Kumari, D.; Sohail, M.; Jahangeer, S.; Abideen, Z.; Khan, M.A.

    2017-01-01

    Fungi elaborate a variety of plant-hydrolyzing enzymes including cellulases, xylanases, pectinases and amylases. Although these enzymes have potential biotechnological applications, their production at industrial level is limited because of higher costs of the purified substrates. Hence, the present study was aimed to explore the novel, natural and cheaper substrates for enzyme production. Indigenously isolated fungal strains of Aspergillus sp. were grown on banana-peels, grapefruit-peels, pomegranate-peels, sugarcane bagasse, Eucalyptus camaldulensis-leaves and shoots of two halophytic plants including Halopyrum mucronatum and Desmostachya bipinnata under solid-state fermentation (SSF) and submerged fermentation (Smf) conditions. The crude enzyme preparation was screened for cellulase (endoglucanase, beta-glucosidase and filter-paperase), hemicellulase (xylanase), pectinase and amylase production. The results revealed that among all investigated enzymes, the xylanase titers were highest using D. bipinnata- shoots and H. mucronatum- shoots as substrates under solid state fermentation conditions, suggesting their exploitation at commercial scale. (author)

  19. Profiling adrenal 11β-hydroxyandrostenedione metabolites in prostate cancer cells, tissue and plasma: UPC2-MS/MS quantification of 11β-hydroxytestosterone, 11keto-testosterone and 11keto-dihydrotestosterone.

    Science.gov (United States)

    du Toit, Therina; Bloem, Liezl M; Quanson, Jonathan L; Ehlers, Riaan; Serafin, Antonio M; Swart, Amanda C

    2017-02-01

    Adrenal C 19 steroids serve as precursors to active androgens in the prostate. Androstenedione (A4), 11β-hydroxyandrostenedione (11OHA4) and 11β-hydroxytestosterone (11OHT) are metabolised to potent androgen receptor (AR) agonists, dihydrotestosterone (DHT), 11-ketotestosterone (11KT) and 11-ketodihydrotestosterone (11KDHT). The identification of 11OHA4 metabolites, 11KT and 11KDHT, as active androgens has placed a new perspective on adrenal C11-oxy C 19 steroids and their contribution to prostate cancer (PCa). We investigated adrenal androgen metabolism in normal epithelial prostate (PNT2) cells and in androgen-dependent prostate cancer (LNCaP) cells. We also analysed steroid profiles in PCa tissue and plasma, determining the presence of the C 19 steroids and their derivatives using ultra-performance liquid chromatography (UHPLC)- and ultra-performance convergence chromatography tandem mass spectrometry (UPC 2 -MS/MS). In PNT2 cells, sixty percent A4 (60%) was primarily metabolised to 5α-androstanedione (5αDIONE) (40%), testosterone (T) (10%), and androsterone (AST) (10%). T (30%) was primarily metabolised to DHT (10%) while low levels of A4, 5αDIONE and 3αADIOL (≈20%) were detected. Conjugated steroids were not detected and downstream products were present at <0.05μM. Only 20% of 11OHA4 and 11OHT were metabolised with the former yielding 11keto-androstenedione (11KA4), 11KDHT and 11β-hydroxy-5α-androstanedione (11OH-5αDIONE) and the latter yielding 11OHA4, 11KT and 11KDHT with downstream products <0.03μM. In LNCaP cells, A4 (90%) was metabolised to AST-glucuronide via the alternative pathway while T was detected as T-glucuronide with negligible conversion to downstream products. 11OHA4 (80%) and 11OHT (60%) were predominantly metabolised to 11KA4 and 11KT and in both assays more than 50% of 11KT was detected in the unconjugated form. In tissue, we detected C11-oxy C 19 metabolites at significantly higher levels than the C 19 steroids, with

  20. Optimization of a synthetic mixture composed of major Trichoderma reesei enzymes for the hydrolysis of steam-exploded wheat straw

    Directory of Open Access Journals (Sweden)

    Billard Hélène

    2012-02-01

    Full Text Available Abstract Background An efficient hydrolysis of lignocellulosic substrates to soluble sugars for biofuel production necessitates the interplay and synergistic interaction of multiple enzymes. An optimized enzyme mixture is crucial for reduced cost of the enzymatic hydrolysis step in a bioethanol production process and its composition will depend on the substrate and type of pretreatment used. In the present study, an experimental design was used to determine the optimal composition of a Trichoderma reesei enzyme mixture, comprising the main cellulase and hemicellulase activities, for the hydrolysis of steam-exploded wheat straw. Methods Six enzymes, CBH1 (Cel7a, CBH2 (Cel6a, EG1 (Cel7b, EG2 (Cel5a, as well as the xyloglucanase Cel74a and the xylanase XYN1 (Xyl11a were purified from a T. reesei culture under lactose/xylose-induced conditions. Sugar release was followed in milliliter-scale hydrolysis assays for 48 hours and the influence of the mixture on initial conversion rates and final yields is assessed. Results The developed model could show that both responses were strongly correlated. Model predictions suggest that optimal hydrolysis yields can be obtained over a wide range of CBH1 to CBH2 ratios, but necessitates a high proportion of EG1 (13% to 25% which cannot be replaced by EG2. Whereas 5% to 10% of the latter enzyme and a xylanase content above 6% are required for highest yields, these enzymes are predicted to be less important in the initial stage of hydrolysis. Conclusions The developed model could reliably predict hydrolysis yields of enzyme mixtures in the studied domain and highlighted the importance of the respective enzyme components in both the initial and the final hydrolysis phase of steam-exploded wheat straw.

  1. Rapid labelling of radiopharmaceuticals using 11CO2 and 11CH4

    International Nuclear Information System (INIS)

    Crouzel, C.

    1988-07-01

    In the past two decades, much effort has been devoted to the development of new molecules, labelled with β+ emitters usable for Positron Emission Tomography. Gaseous forms of 11 C ( 11 CO 2 or 11 CH 4 ) must be converted to a reactive form known as a ''radioactive precursor'': 11 C-methanol, 11 C-formaldehyde, 11 C-acetone, 11 C-phosgene, 11 C-diazomethane, 11 C-methylamine. These precursors are used to label radiopharmaceuticals. Few examples are given: 11 C-prazosin, 11 C-CGP 12177, 11 C-pindolol. Such synthesis procedures require strong initial activity (1.5 Ci). The processes are therefore remotely controlled or automated, and confined to shielded cells. Small laboratory robots have lately been introduced for this type of production

  2. Enhancement in catalytic activity of Aspergillus niger XynB by selective site-directed mutagenesis of active site amino acids.

    Science.gov (United States)

    Wu, Xiuyun; Tian, Zhennan; Jiang, Xukai; Zhang, Qun; Wang, Lushan

    2018-01-01

    XynB from Aspergillus niger ATCC1015 (AnXynB) is a mesophilic glycoside hydrolase (GH) family 11 xylanase which holds great potentials in a wide variety of industrial applications. In the present study, the catalytic activity and stability of AnXynB were improved by a combination of computational and experimental approaches. Virtual mutation and molecular dynamics simulations indicated that the introduction of Glu and Asn altered the interaction network at the - 3 subsite. Interestingly, the double mutant S41N/T43E displayed 72% increase in catalytic activity when compared to the wild type (WT). In addition, it also showed a better thermostability than the WT enzyme. Kinetic determination of the T43E and S41N/T43E mutants suggested that the higher xylanase activity is probably due to the increasing binding affinity of enzyme and substrate. Consequently, the enzyme activity and thermostability of AnXynB was both increased by selective site-directed mutagenesis at the - 3 subsite of its active site architecture which provides a good example for a successfully engineered enzyme for potential industrial application. Moreover, the molecular evolution approach adopted in this study led to the design of a library of sequences that captures a meaningful functional diversity in a limited number of protein variants.

  3. Ubiquitin-specific protease 11 (USP11) functions as a tumor suppressor through deubiquitinating and stabilizing VGLL4 protein

    Science.gov (United States)

    Zhang, Encheng; Shen, Bing; Mu, Xingyu; Qin, Yan; Zhang, Fang; Liu, Yong; Xiao, Jiantao; Zhang, Pingzhao; Wang, Chenji; Tan, Mingyue; Fan, Yu

    2016-01-01

    VGLL4 is a transcriptional repressor that interacts with transcription factors TEADs and inhibits YAP-induced overgrowth and tumorigenesis. VGLL4 protein was dramatically reduced in various types of human cancers. But how VGLL4 protein is post-transcriptional regulated is poorly understood. In this study, we identify deubiquitinating enzyme USP11 as a novel VGLL4 interactor. We reveal that the USP domain of USP11 and the N-terminal region of VGLL4 are required for mutual binding. USP11 controls VGLL4 protein stability by promoting its deubiquitination. Furthermore, our results show that knockdown of USP11 promotes cell growth, migration, and invasion in a YAP-dependent manner. Together, our results suggest that USP11 may exert its tumor suppressor role by modulating VGLL4/YAP-TEADs regulatory loop. PMID:28042509

  4. Evidence of a new state in 11Be observed in the 11Li β-decay

    International Nuclear Information System (INIS)

    Madurga, M.; Borge, M.J.G.; Alcorta, M.; Fraile, L.M.; Fynbo, H.O.U.; Jonson, B.; Kirsebom, O.; Martinez-Pinedo, G.; Nilsson, T.; Nyman, G.; Perea, A.; Poves, A.; Riisager, K.; Tengblad, O.; Tengborn, E.; Van der Walle, J.

    2009-01-01

    Coincidences between charged particles emitted in the β-decay of 11 Li were observed using highly segmented detectors. The breakup channels involving three particles were studied in full kinematics allowing for the reconstruction of the excitation energy of the 11 Be states participating in the decay. In particular, the contribution of a previously unobserved state at 16.3 MeV in 11 Be has been identified selecting the α+ 7 He→α+ 6 He+n channel. The angular correlations between the α particle and the center of mass of the 6 He + n system favors spin and parity assignment of 3/2 - for this state as well as for the previously known state at 18 MeV.

  5. Screening and Molecular Identification of New Microbial Strains for Production of Enzymes of Biotechnological Interest

    Directory of Open Access Journals (Sweden)

    Imen Ghazala

    Full Text Available ABSTRACT: This research focused on isolation, identification and characterization of new strains of fungi and bacteria, which were able to produce extracellular xylanase, mannanase, pectinase and α-amylase. Fungi isolates were identified on the basis of analyses of 18S gene sequencing and internal transcribed spacer region. The closest phylogenetic neighbors according to 18S gene sequence and ITS region data for the two isolates M1 and SE were Aspergillus fumigatus and Aspergillus sydowii, respectively. I4 was identified as Bacillus mojavensis on the basis of the 16S rRNA gene sequencing and biochemical properties. The enzyme production was evaluated by cultivating the isolated microorganisms in liquid-state bioprocess using wheat bran as carbon source. Two fungi (M1, and SE and one bacterium (I4 strains were found to be xylanase producer, and several were proven to be outstanding producers of microbial xylanase. The strains producing xylanase secreted variable amounts of starch-debranching enzymes and produced low level β-mannan-degrading enzyme systems. The bacterium strain was found to be capable of producing pectinolytic enzymes on wheat bran at high level. Some of the strains have good potential for use as sources of important industrial enzymes.

  6. Purification and biochemical properties of a new thermostable ...

    African Journals Online (AJOL)

    The enzyme was stable for a long time-period up to 50°C and for 1 h at 60°C. Although the xylanase had a lower carboxymethylcellulase activity, it lacked activity towards substituted xylan, xylobiose, inulin, starch, polygalacturonic acid or pNP glycosides. Kinetic parameters indicated higher efficiency in the hydrolysis of ...

  7. DETERMINATION OF ENZYMES PRODUCED BY CERIPORIOPSIS SUBVERMISPORA DURING PRETREATMENT OF DIFFERENT BIOMASS SOURCES

    Directory of Open Access Journals (Sweden)

    Miroslav Ondrejovič

    2012-02-01

    Full Text Available The aim of this paper was to study of lignocellulolytic enzymes producing by Ceriporiopsis subvermispora during its cultivation on three types of plant biomass differentiated by chemical composition and physical properties (wheat straw, pine and poplar wood. The activity of lignocellulolytic enzymes in cultivation medium was determined by catalytic transformation of their natural substrates to products which were detected by photometric methods. Cellulase activities were very low while xylanases predominated. Wheat straw was best substrate for production of cellulases (4.38 U/mL and xylanases (23.34 U/mL. The maximum activity of cellulase and xylanase was reached at 8th and 3rd day, respectively. Laccase activity reached the maximum after 16 days and then gradually decreased. The best substrate for production of laccases was poplar wood (1.67 U/mL.

  8. Production and immobilization of enzymes by solid-state fermentation of agroindustrial waste.

    Science.gov (United States)

    Romo Sánchez, Sheila; Gil Sánchez, Irene; Arévalo-Villena, María; Briones Pérez, Ana

    2015-03-01

    The recovery of by-products from agri-food industry is currently one of the major challenges of biotechnology. Castilla-La Mancha produces around three million tons of waste coming from olive oil and wine industries, both of which have a pivotal role in the economy of this region. For this reason, this study reports on the exploitation of grape skins and olive pomaces for the production of lignocellulosic enzymes, which are able to deconstruct the agroindustrial waste and, therefore, reuse them in future industrial processes. To this end, solid-state fermentation was carried out using two local fungal strains (Aspergillus niger-113 N and Aspergillus fumigatus-3). In some trials, a wheat supplementation with a 1:1 ratio was used to improve the growth conditions, and the particle size of the substrates was altered through milling. Separate fermentations were run and collected after 2, 4, 6, 8, 10 and 15 days to monitor enzymatic activity (xylanase, cellulase, β-glucosidase, pectinase). The highest values were recorded after 10 and 15 days of fermentation. The use of A. niger on unmilled grape skin yielded the best outcomes (47.05 U xylanase/g by-product). The multi-enzymatic extracts obtained were purified, freeze dried, and immobilized on chitosan by adsorption to assess the possible advantages provided by the different techniques.

  9. Local E11

    International Nuclear Information System (INIS)

    Riccioni, Fabio; West, Peter

    2009-01-01

    We give a method of deriving the field-strengths of all massless and massive maximal supergravity theories in any dimension starting from the Kac-Moody algebra E 11 . Considering the subalgebra of E 11 that acts on the fields in the non-linear realisation as a global symmetry, we show how this is promoted to a gauge symmetry enlarging the algebra by the inclusion of additional generators. We show how this works in eleven dimensions, and we call the resulting enlarged algebra E 11 local . Torus reduction to D dimensions corresponds to taking a subalgebra of E 11 local , called E 11,D local , that encodes the full gauge algebra of the corresponding D-dimensional massless supergravity. We show that each massive maximal supergravity in D dimensions is a non-linear realisation of an algebra E-tilde 11,D local . We show how this works in detail for the case of Scherk-Schwarz reduction of IIB to nine dimensions, and in particular we show how E-tilde 11,9 local arises as a subalgebra of the algebra E 11,10B local associated to the ten-dimensional IIB theory. This subalgebra corresponds to taking a combination of generators which is different to the massless case. We then show that E-tilde 11,D local appears as a deformation of the massless algebra E 11,D local in which the commutation relations between the E 11 and the additional generators are modified. We explicitly illustrate how the deformed algebra is constructed in the case of massive IIA and of gauged five-dimensional supergravity. These results prove the naturalness and power of the method.

  10. Synthesis of n.c.a. PET-radiotracers with carbon-11

    International Nuclear Information System (INIS)

    Schirbel, A.

    1998-11-01

    Carbon-11 offers the unique possibility of authentic labelling of molecules as radioindicators for non invasive and quantitative determination of physiological functions via positron emission tomography (PET). Therefore, the goal of this thesis was to synthesize of different n.c.a. 11 C-labelled pharmaceuticals for in vivo distribution studies with PET. For the determination of the pharmacokinetics of [1- 11 C]acetate in porcine myocardium during prolonged ischemia, n.c.a. [1- 11 C]acetate was synthesized via carboxylation of methylmagnesium bromide with in target produced n.c.a. [ 11 C]CO 2 with a radiochemical yield (RCY) of 68 ± 7%. The fast (18 min) and reliable radiosynthesis allowed for repeated tracer administration at short intervals ( R c) in humans, [1- 11 C]acetylsalicylic acid, acetyl-[carboxy- 11 C]salicylic acid and [carboxy- 11 C]salicylic acid were prepared. N.c.a. [1- 11 C]acetylsalicylic acid was synthesized via the reaction of [1- 11 C]acetylchloride with salicylic acid salts. The use of the silver salt proved to be superior to the sodium salt and resulted in radiochemical yields of 32 ± 5%. Base-line (clean) separation of the labelled product was achieved using radio-HPLC. With regard to the preparation of n.c.a. [carboxy- 11 C]salicylic acid, several protected and unprotected phenol derivates were metallated and subsequently carboxylated using n.c.a. [ 11 C]CO 2 . Best results (87 ± 3% RCY) could be achieved with 2-(methoxymethoxy)-phenylmagnesium iodide as a precursor and subsequent quantitative cleavage of the MOM-group. Acetylation of n.c.a. [carboxy- 11 C]salicylic acid to acetyl-[carboxy- 11 C]salicylic acid was performed using acetylchloride in CH 2 Cl 2 with a radiochemical yield of 65 ± 4%. (orig.)

  11. Biosynthesis of cellulolytic enzymes by Aspergillus niger A. n. 33 and its selectants

    Energy Technology Data Exchange (ETDEWEB)

    Czajkowska, D.; Hornecka, D.; Ilnicka-Olejniczak, O.

    1988-01-01

    The aim of the investigations was to obtain - from the parent strain Aspergillus niger A.n. 33 - selectants with an increased ability of cellulolytic enzymes biosynthesis. Own selection methods allowed to receive two selectants A.n. 33/2 and A.n. 33/20 characterized by enhanced activities of saccharifying cellulase (respectively 0.11 and 0.14 FPU/cm/sup 3/), endo-beta-1,4-glucanase (15.4 and 21.8 U/cm/sup 3/) and cellobiase (0.6 and 1.4 IU/cm/sup 3/) as compared with the parent strain (FPA - 0.09 IU, CMC - 8.2 U and CB - 0.1 IU/cm/sup 3/). Moreover, the selectants differed in shape and size of conidial heads, in shape and colour of conidia, as well as in structure and shape of hyphase. Enzyme preparations obtained after ultrafiltration of liquid cultures were characterized by following activities: FPA-4-16 IU, CMC-900-1800 U, CB-60-120 IU and xylanase-250-280 IU/cm/sup 3/.

  12. Binding kinetics of 11C-N-methyl piperidyl benzilate (11C-NMPB) in a rhesus monkey brain using the cerebellum as a reference region

    International Nuclear Information System (INIS)

    Itoh, Takashi; Tanaka, Masayasu; Suzuki, Kasutoshi; Kobayashi, Kaoru; Inoue, Osamu

    2005-01-01

    The binding kinetics of' 11 C-N-methyl piperidyl benzilate ( 11 C-NMPB) in rhesus monkey brain were studied using animal positron emission tomography (PET) (SHR2000). This study is intended to assess the validity of the method using the cerebellum as a reference region, and to evaluate the effects of anesthesia on 11 C -NMPB binding. Two monkeys, anesthetized with ketamine, received intravenous 11 C-NMPB alone (370-760 MBq, 11 C-NMPB accumulated densely in the striatum and cerebral cortex with time. In contrast, the tracer accumulation significantly decreased with increased doses of nonradioactive NMPB. In the cerebellum, on the other hand, the accumulation of 11 C-NMPB remained low and the tracer was slowly eliminated from the brain following the injection. 11 C-NMPB binding in the cerebellum was barely affected by the increased dose of nonradioactive NMPB. We thus concluded that the specific 11 C-NMPB binding was negligible in the cerebellum, and performed simplified evaluation of 11 C-NMPB binding in each brain region by a graphical method using the cerebellum as a reference region. PET was conducted 26 times, in total both in ketamine-anesthetized and awake monkeys (n=3 each). Measurements of 11 C-NMPB binding showed good run-to-run reproducibility within individual animals. When 11 C-NMPB binding was compared between ketamine-treated and awake animals, a significant increase in 11 C-NMPB binding was observed in the striatum but not in other brain regions of ketamine-treated animals. (author)

  13. Catalytic conversion of 11CO2 and 11CO into synthesis precursors for 11C labelling

    International Nuclear Information System (INIS)

    Patt, J.T.

    1994-03-01

    The positron emitter carbon-11 (T 1/2 =20.3 min) is an ideal radio nuclide for tracers in positron emission tomography (PET). In this study catalytic methods for the synthesis of [ 11 C]alcohols have been investigated. The formation of [ 11 C]methanol has been studied on Pd/Al 2 O 3 and Cu/ZnO/Al 2 O 3 catalysts with respect to CO and CO 2 carrier addition to the synthesis gas. Carbon monoxide was identified as the precursor of methanol formation on the Pd/Al 2 O 3 -catalyst. In contrast on the Cu/ZnO/Al 2 O 3 -catalyst methanol was formed on a reaction pathway via an adsorbed CO 2 -species. A n.c.a.-[ 11 C]methanol synthesis basing on the Cu/ZnO/Al 2 O 3 -catalyst has been developed by substitution of the oxygen containing components CO and CO 2 in the synthesis gas by N 2 O. The radiochemical yield, the low selectivity of [ 11 C]methanol production and the rather slow kinetics of this process were arguments against the practical use of this process in the synthesis of 11 C-labelling agents. (orig.)

  14. Rapid screening and structural elucidation of a novel sibutramine analogue in a weight loss supplement: 11-desisobutyl-11-benzylsibutramine.

    Science.gov (United States)

    Mans, Daniel J; Gucinski, Ashley C; Dunn, Jamie D; Gryniewicz-Ruzicka, Connie M; Mecker-Pogue, Laura C; Kao, Jeff L-F; Ge, Xia

    2013-09-01

    A novel analogue of sibutramine, 11-desisobutyl-11-benzylsibutramine, has been discovered. During routine ion mobility spectrometry (IMS) screening of a weight loss supplement collected at an US FDA import operation facility an unknown peak was observed. Further analysis of the supplement by liquid chromatography-mass spectrometry (LC-MS) and high resolution mass spectrometry revealed an unknown peak with a relative retention time of 1.04 with respect to sibutramine and a predicted formula of C20H24NCl. In order to elucidate the analogue's structure, it was isolated from the supplement and characterized by tandem mass spectrometry and nuclear magnetic resonance (NMR), which revealed the analogue possessed a benzyl moiety at the 11 position in place of the isobutyl group associated with sibutramine. Copyright © 2013. Published by Elsevier B.V.

  15. Carbon-11 labelling of an inhibitor of acetylcholinesterase: [11C]physostigmine

    International Nuclear Information System (INIS)

    Bonnot-Lours, S.; Crouzel, C.; Prenant, C.; Hinnen, F.

    1993-01-01

    Physostigmine, an alkaloid from calabar bean is a strong inhibitor of acetylcholinesterase and has been used clinically in the treatment of glaucoma, atropine intoxication, myasthenia gravis and more recently, in experimental trials in Alzheimer's disease. In order to study the AChE activity in the brain by positron emission tomography, we have undertaken the labelling of physostigmine with carbon-11. The synthesis involves the reaction of [ 11 C]methylisocyanate with eseroline. [ 11 C]Methylisocyanate was obtained by heating [ 11 C]acetylchloride with tetrabutylammonium azide in toluene. The synthesis of [ 11 C]CH 3 COC1 involves the carbonation of methylmagnesium bromide in THF with cyclotron produced [ 11 C]carbon dioxide and the addition of phthaloyl dichloride. The [ 11 C]methylisocyanate is distilled into a solution of eseroline in ether with a small piece of sodium. After 10 minutes at 25 o C, the solution is purified by HPLC and the appropriate fraction collected. Starting with 55.5 GBq (1.5 Ci) of [ 11 C]carbon dioxide, 0.92-1.48 GBq (25-40 mCi) of [ 11 C]Physostigmine are obtained 57 minutes after EOB. (author)

  16. 11-Deoxycortisol is a corticosteroid hormone in the lamprey

    Science.gov (United States)

    Close, D.A.; Yun, S.-S.; McCormick, S.D.; Wildbill, A.J.; Li, W.

    2010-01-01

    Corticosteroid hormones are critical for controlling metabolism, hydromineral balance, and the stress response in vertebrates. Although corticosteroid hormones have been well characterized in most vertebrate groups, the identity of the earliest vertebrate corticosteroid hormone has remained elusive. Here we provide evidence that 11-deoxycortisol is the corticosteroid hormone in the lamprey, a member of the agnathans that evolved more than 500 million years ago. We used RIA, HPLC, and mass spectrometry analysis to determine that 11-deoxycortisol is the active corticosteroid present in lamprey plasma. We also characterized an 11-deoxycortisol receptor extracted from sea lamprey gill cytosol. The receptor was highly specific for 11-deoxycortisol and exhibited corticosteroid binding characteristics, including DNA binding. Furthermore, we observed that 11-deoxycortisol was regulated by the hypothalamus-pituitary axis and responded to acute stress. 11-Deoxycortisol implants reduced sex steroid concentrations and upregulated gill Na+, K+-ATPase, an enzyme critical for ion balance. We show here that 11-deoxycortisol functioned as both a glucocorticoid and a mineralocorticoid in the lamprey. Our findings indicate that a complex and highly specific corticosteroid signaling pathway evolved at least 500 million years ago with the arrival of the earliest vertebrate.

  17. Synthesis of racemic [3-11C]phenylalanine and [3-11C] DOPA

    International Nuclear Information System (INIS)

    Halldin, C.; Laangstroem, B.

    1984-01-01

    The synthesis of racemic [3- 11 C]phenylalanine and [3- 11 C]DOPA is reported. The [ 11 C]benzaldehyde and [ 11 C]veratraldehyde prepared in a two-step reaction from the corresponding [ 11 C]acid salt and [ 11 C]alcohol, by means of selective oxidation with tetrabutylammonium hydrogen chromate, were reacted with 2-phenyl-5-oxazolone or 2-(4-chloro)phenyl-5-oxazolone in the presence of a tertiary amine to give the corresponding [α- 11 C]-4-arylene-2-aryl-5-oxazolones. Ring opening of these olefins, hydrogenation, and removal of protecting groups was carried out in one step using hydroiodic cid/phosphorus, with the production of the racemic [3- 11 C]amino acids in 8-30% radiochemical yield (starting with 11 CO 2 ) within 52-60 min (including LC separation). (author)

  18. In vivo evaluation of alpha7 nicotinic acetylcholine receptor agonists [11C]A-582941 and [11C]A-844606 in mice and conscious monkeys.

    Directory of Open Access Journals (Sweden)

    Jun Toyohara

    Full Text Available BACKGROUND: The alpha7 nicotinic acetylcholine receptors (nAChRs play an important role in the pathophysiology of neuropsychiatric diseases such as schizophrenia and Alzheimer's disease. The goal of this study was to evaluate the two carbon-11-labeled alpha7 nAChR agonists [(11C]A-582941 and [(11C]A-844606 for their potential as novel positron emission tomography (PET tracers. METHODOLOGY/PRINCIPAL FINDINGS: The two tracers were synthesized by methylation of the corresponding desmethyl precursors using [(11C]methyl triflate. Effects of receptor blockade in mice were determined by coinjection of either tracer along with a carrier or an excess amount of a selective alpha7 nAChR agonist (SSR180711. Metabolic stability was investigated using radio-HPLC. Dynamic PET scans were performed in conscious monkeys with/without SSR180711-treatment. [(11C]A-582941 and [(11C]A-844606 showed high uptake in the mouse brain. Most radioactive compounds in the brain were detected as an unchanged form. However, regional selectivity and selective receptor blockade were not clearly observed for either compound in the mouse brain. On the other hand, the total distribution volume of [(11C]A-582941 and [(11C]A-844606 was high in the hippocampus and thalamus but low in the cerebellum in the conscious monkey brain, and reduced by pretreatment with SSR180711. CONCLUSIONS/SIGNIFICANCE: A nonhuman primate study suggests that [(11C]A-582941 and [(11C]A-844606 would be potential PET ligands for imaging alpha7 nAChRs in the human brain.

  19. A Comparative Study of Temperature Optimal Control in a Solid State Fermentation Process for Edible Mushroom Growing

    Directory of Open Access Journals (Sweden)

    K. J. Gurubel

    2017-04-01

    Full Text Available In this paper, optimal control strategies for temperature trajectory determination in order to maximize thermophilic bacteria in a fed-batch solid-state fermentation reactor are proposed. This process is modeled by nonlinear differential equations, which has been previously validated experimentally with scale reactor temperature profiles. The dynamic input aeration rate of the reactor is determined to increase microorganisms growth of a selective substrate for edible mushroom cultivation. In industrial practice, the process is comprised of three thermal stages with constant input air flow and three types of microorganisms in a 150-hour lapse. Scytalidium thermophilum and actinobacteria are desired in order to obtain a final biomass composition with acceptable microorganisms concentration. The Steepest Descent gradient algorithm in continuous time and the Gradient Projection algorithm in discrete-time are used for the process optimal control. A comparison of simulation results in the presence of disturbances is presented, where the resulting temperature trajectories exhibit similar tendencies as industrial data.

  20. 26 CFR 11.412(c)-11 - Election with respect to bonds.

    Science.gov (United States)

    2010-04-01

    ... amount payable at maturity (or, in the case of a bond which is callable prior to maturity, the earliest... 26 Internal Revenue 14 2010-04-01 2010-04-01 false Election with respect to bonds. 11.412(c)-11... OF 1974 § 11.412(c)-11 Election with respect to bonds. (a) In general. Section 412(c)(2)(B) provides...

  1. Use of a human chromosome 11 radiation hybrid panel to map markers at 11q13

    International Nuclear Information System (INIS)

    Withers, D.; Richard, C. III; Meeker, T.C.; Maurer, S.; Evans, G.; Myers, R.M.; Cox, D.R.

    1990-01-01

    A human/hamster hybrid cell line containing human chromosome 11 was X-irradiated and 102-independent derivative lines were recovered. These 'radiation hybrids' contain random fragments of human chromosome 11. This radiation hybrid panel was used to score the retention of markers at band 11q13. Statistical analysis of marker co-retention patterns in the radiation hybrid panel permits a preliminary ordering and mapping of the markers used. The best order for six scored markers is: proximal - CD5 - CD20 - PGA - HST - BCL1 - SEA - distal. Additional markers are currently being scored. The six 11q13 markers above are spread over approximately 10-12 mB of DNA. The mapping data has implications for the identification of the bcl-1 gene. bcl-1 is the site of chromosome breakage in translocations associated with B lymphocytic malignancy. bcl-1 markers map at least 4 Mb away from any of four genes previously hypothesized to be activated by such translocations, thereby making them unlikely candidates for activation

  2. Dendritic cell fate is determined by BCL11A

    Science.gov (United States)

    Ippolito, Gregory C.; Dekker, Joseph D.; Wang, Yui-Hsi; Lee, Bum-Kyu; Shaffer, Arthur L.; Lin, Jian; Wall, Jason K.; Lee, Baeck-Seung; Staudt, Louis M.; Liu, Yong-Jun; Iyer, Vishwanath R.; Tucker, Haley O.

    2014-01-01

    The plasmacytoid dendritic cell (pDC) is vital to the coordinated action of innate and adaptive immunity. pDC development has not been unequivocally traced, nor has its transcriptional regulatory network been fully clarified. Here we confirm an essential requirement for the BCL11A transcription factor in fetal pDC development, and demonstrate this lineage-specific requirement in the adult organism. Furthermore, we identify BCL11A gene targets and provide a molecular mechanism for its action in pDC commitment. Embryonic germ-line deletion of Bcl11a revealed an absolute cellular, molecular, and functional absence of pDCs in fetal mice. In adults, deletion of Bcl11a in hematopoietic stem cells resulted in perturbed yet continued generation of progenitors, loss of downstream pDC and B-cell lineages, and persisting myeloid, conventional dendritic, and T-cell lineages. Challenge with virus resulted in a marked reduction of antiviral response in conditionally deleted adults. Genome-wide analyses of BCL11A DNA binding and expression revealed that BCL11A regulates transcription of E2-2 and other pDC differentiation modulators, including ID2 and MTG16. Our results identify BCL11A as an essential, lineage-specific factor that regulates pDC development, supporting a model wherein differentiation into pDCs represents a primed “default” pathway for common dendritic cell progenitors. PMID:24591644

  3. Renal cell carcinoma and a constitutional t(11;22)(q23;q11.2): case report and review of the potential link between the constitutional t(11;22) and cancer.

    Science.gov (United States)

    Doyen, Jérôme; Carpentier, Xavier; Haudebourg, Juliette; Hoch, Benjamin; Karmous-Benailly, Houda; Ambrosetti, Damien; Fabas, Thibault; Amiel, Jean; Lambert, Jean-Claude; Pedeutour, Florence

    2012-11-01

    We observed a t(11;22)(q23-24;q11.2-12) and monosomy 3 in renal tumor cells from a 72-year-old man. The hypothesis of a primitive peripheral neuroectodermal tumor (PPNET) located in the kidney was promptly excluded: Histologically, the tumor was a clear cell renal cell carcinoma (RCC) and we did not observe an EWSR1 gene rearrangement. The constitutional origin of this alteration was established. We report on the second case of RCC in a patient with a constitutional t(11;22). The t(11;22)(q23;q11.2) is the main recurrent germline translocation in humans. Unbalanced translocation can be transmitted to the progeny and can cause Emanuel syndrome. Our observation alerts cancer cytogeneticists to the fortuitous discovery of the constitutional t(11;22) in tumor cells. This translocation appears grossly similar to the t(11;22)(q24;q12) of PPNET and should be evoked if present in all cells of a tumor other than PPNET. This is important when providing appropriate genetic counseling. Moreover, the potential oncogenic role of the t(11;22) and its predisposing risk of cancer are under debate. The family history of the patient revealed a disabled brother who died at an early age from colon cancer and a sister with breast cancer. This observation reopens the issue of a link between the constitutional t(11;22) and cancer, and the utility of cancer prevention workups for t(11;22) carriers. Copyright © 2012 Elsevier Inc. All rights reserved.

  4. Production of 11Li in the (11B,11Li) reaction on 232Th

    International Nuclear Information System (INIS)

    Scott, D.K.; Buenerd, M.; Hendrie, D.L.; KeKelis, G.; Mahoney, J.; Menchaca-Rocha, A.; Olmer, C.

    1975-01-01

    Production of the neutron-rich nucleus 11 Li in the bombardment of 232 Th by 11 B at 114 MeV suggests that multinucleon transfer reactions induced by neutron excess heavy ions on heavy targets present a feasible method of measuring the mass excess of exotic light nuclei in the limit of stability

  5. Radiosynthesis of [11C]D.P.A.-713, [11C]D.P.A.-715 and [11C]clinme, selected carbon-11-labelled novel potential radioligands for imaging the peripheral benzodiazepine receptors with PET

    International Nuclear Information System (INIS)

    Dolle, F.; Thominiaux, C.; Hinnen, F.; Demphel, S.; Le helleix, S.; Chauveau, F.; Boutin, H.; Herard, A.S.; Hantraye, P.; Tavitian, B.; Kassiou, M.; James, M.; Creelman, A.; Fulton, R.; Kassiou, M.; Katsifis, A.; Greguric, I.; Mattner, F.; Loch, C.; Selleri, S.

    2008-01-01

    11 C P.K.11195 is not only the oldest, but also the most widely used PET radiotracer for in vivo imaging of the peripheral benzodiazepine receptors (P.B.R. or translocator protein (18 kDa, T.S.P.O.). With the aim of developing a new PET imaging probe for the in vivo study of the P.B.R., two pyrazol [1,5-a]pyrimidineacetamides (D.P.A.-713 and D.P.A.-715) and one imidazol[1,2-a]pyridine-acetamide (C.L.I.N.M.E.) were radiolabelled with the positron emitters carbon 11 (half life: 20.38 min) [1-5]. Briefly, C.L.I.N.M.E. (2-[6-chloro-2(4-iodophenyl)-imidazol[1,2-a]pyridin-3-yl] -N-ethyl-N-methyl-acetamide) was labelled at its methyl-acetamide moity chain from the corresponding nor-analogue using[ 11 C]methyl iodide (in D.M.S.O./D.M.F (100/200 μL) containing powdered K.O.H. (3-5 mg) at 110 degrees C for 3 min. D.P.A.-713 (N,N-diethyl-2-[2-(4-methoxy-phenyl)-5,7-dimethyl-pyrazolo[1,5-a]pyrimidin -3-yl]acetamide) and D.P.A.-715 (N,N-diethyl-2-[2-(4-methoxy-phenyl)-5,7-bis-tri-fluoro-methyl-pyrazolo [1,5-a]pyrimidin-3-yl]acetamide) were labelled at their aromatic methoxy groups from the corresponding nor-derivatives using [ 11 C]methyl triflate (in acetone (300μL) containing aq. 3 M NaOH (4μL) at 110 degrees C for 1 min). All radioligands were purified using semi preparative Zorbax reverse phase H.P.L.C., were adequately formulated for in vivo injection within 30 min and were found to be > 95% chemically and radiochemically pure. (N.C.)

  6. The ACEII recombinant Trichoderma reesei QM9414 strains with enhanced xylanase production and its applications in production of xylitol from tree barks.

    Science.gov (United States)

    Xiong, Lili; Kameshwar, Ayyappa Kumar Sista; Chen, Xi; Guo, Zhiyun; Mao, Canquan; Chen, Sanfeng; Qin, Wensheng

    2016-12-28

    ACEII transcription factor plays a significant role in regulating the expression of cellulase and hemicellulase encoding genes. Apart from ACEII, transcription factors such as XYR1, CRE1, HAP2/3/5 complex and ACEI function in a coordinated pattern for regulating the gene expression of cellulases and hemicellulases. Studies have demonstrated that ACEII gene deletion results in decreased total cellulase and xylanase activities with reduced transcript levels of lignocellulolytic enzymes. In this study, we have successfully transformed the ACEII transcription factor encoding gene in Trichoderma reesei to significantly improve its degrading abilities. Transformation experiments on parental strain T. reesei QM9414 has resulted in five genetically engineered strains T/Ace2-2, T/Ace2-5, T/Ace2-8, T/Ace5-4 and T/Ace10-1. Among which, T/Ace2-2 has exhibited significant increase in enzyme activity by twofolds, when compared to parental strain. The T/Ace2-2 was cultured on growth substrates containing 2% bark supplemented with (a) sugar free + MA medium (b) glucose + MA medium and (c) xylose + MA medium. The bark degradation efficiency of genetically modified T/Ace2-2 strain was assessed by analyzing the xylitol production yield using HPAEC. By 6th day, about 10.52 g/l of xylitol was produced through enzymatic conversion of bark (2% bark + MA + xylose) by the T/Ace2-2 strain and by 7th day the conversion rate was found to be 0.21 g/g. Obtained results confirmed that bark growth medium supplemented with D-xylose has profoundly increased the conversion rate of bark by T/Ace2-2 strain when compared to sugar free and glucose supplemented growth media. Results obtained from scanning electron microscopy has endorsed our current results. Bark samples inoculated with T/Ace2-2 strain has showed large number of degraded cells with clearly visible cavities and fractures, by exposing the microfibrillar interwoven complex. We propose a cost effective and ecofriendly method for

  7. Teaching through a Crisis: September 11 and Beyond. Occasional Paper Series 11

    Science.gov (United States)

    McKersie, Alison; Lent, Patricia; Rose, Megan; Stuart-Hunt, Rella; Edstrom, Lisa; Rothschild, Cynthia R.; Weisman, Alexandra; Delacorte, Kate; Vascellaro, Sal; Abell, Kate

    2003-01-01

    The days following September 11, 2001, were characterized for many by a startling clarity, as if someone had applied a magnified lens to life's everyday routines and every moment, every gesture, every word exchanged with friends, colleagues and family was more nuanced and precious than before. The decision to publish this volume of essays was…

  8. Effects of Two-stage Controlled pH and Temperature vs. One-step Process for Hemicellulase Biosynthesis and Feruloyl Oligosaccharide Fermentation using Aureobasidium pullulans

    Directory of Open Access Journals (Sweden)

    Xiaohong Yu

    2016-04-01

    Full Text Available A two-stage, pH- and temperature-controlled wheat bran fermentation method using Aureobasidium pullulans was investigated for feruloyl oligosaccharides (FOs production and the activities of xylanase, xylosidase, and ferulic acid esterase (FAE. A. pullulans secreted xylanase, xylosidase, and FAE at high levels in the initial pH of 4.0 to 5.0 and a fermentation liquid temperature of 31 °C to 33 °C. FOs production via two-stage fermentation (FOs 2 reached 1123 nmol/L after fermentation for 96 h, by controlling the initial pH at 4.0 and the initial temperature at 33 °C, and then changing the pH to 6.0 and the temperature to 29 °C at the same time at 36 h. This process was 12 h shorter and 219 nmol/L higher than a one-stage fermentation for producing FOs 1. Xylanase, xylosidase, and FAE activities were highly correlated with controlled pH and temperature and FOs biosynthesis rate. Thus, the combination of two-stage controlled pH and temperature could support mass production of FOs.

  9. Effects of Designer Hyper-Interleukin 11 (H11 on Hematopoiesis in Myelosuppressed Mice.

    Directory of Open Access Journals (Sweden)

    Hanna Dams-Kozlowska

    Full Text Available The incidence of cancer is constantly increasing. Chemo/radiotherapy is one of major methods of treating cancer. Although adverse chemo/radiotherapy events, such as anemia and neutropenia, can be successfully cured, thrombocytopenia is still problematic. We constructed the Hyper-IL11 (H11 cytokine by linking soluble interleukin 11 receptor alpha (sIL-11Ralpha with IL-11. In vivo H11 activity was examined in myelosuppressed mice. Myelosuppression was induced by either i sublethal irradiation and carboplatin administration or ii sublethal irradiation. A dose of 100 μg/kg of H11 or IL-11 was administered subcutaneously for 7 days. IL-11 and H11 accelerated leukocyte, hematocrit and platelet recovery. The effect on the attenuation of thrombocytopenia was significant. Moreover, both cytokines increased the cellularity and numbers of megakaryocyte, erythroid, and granulocyte/macrophage progenitors in the bone morrow and spleen compared with the control. Although H11 was administered at a molar concentration that was three times lower, its effects were comparable with or better than those of IL-11; thus, the activity of H11 was superior to that of IL-11. Because no toxicity was observed after the intravenous administration of H11, this hyper-cytokine may be potentially useful for treatment of thrombocytopenia and other IL-11-dependent disorders.

  10. Munc13-4 Is a Rab11-binding Protein That Regulates Rab11-positive Vesicle Trafficking and Docking at the Plasma Membrane.

    Science.gov (United States)

    Johnson, Jennifer L; He, Jing; Ramadass, Mahalakshmi; Pestonjamasp, Kersi; Kiosses, William B; Zhang, Jinzhong; Catz, Sergio D

    2016-02-12

    The small GTPase Rab11 and its effectors control trafficking of recycling endosomes, receptor replenishment and the up-regulation of adhesion and adaptor molecules at the plasma membrane. Despite recent advances in the understanding of Rab11-regulated mechanisms, the final steps mediating docking and fusion of Rab11-positive vesicles at the plasma membrane are not fully understood. Munc13-4 is a docking factor proposed to regulate fusion through interactions with SNAREs. In hematopoietic cells, including neutrophils, Munc13-4 regulates exocytosis in a Rab27a-dependent manner, but its possible regulation of other GTPases has not been explored in detail. Here, we show that Munc13-4 binds to Rab11 and regulates the trafficking of Rab11-containing vesicles. Using a novel Time-resolved Fluorescence Resonance Energy Transfer (TR-FRET) assay, we demonstrate that Munc13-4 binds to Rab11a but not to dominant negative Rab11a. Immunoprecipitation analysis confirmed the specificity of the interaction between Munc13-4 and Rab11, and super-resolution microscopy studies support the interaction of endogenous Munc13-4 with Rab11 at the single molecule level in neutrophils. Vesicular dynamic analysis shows the common spatio-temporal distribution of Munc13-4 and Rab11, while expression of a calcium binding-deficient mutant of Munc13-4 significantly affected Rab11 trafficking. Munc13-4-deficient neutrophils showed normal endocytosis, but the trafficking, up-regulation, and retention of Rab11-positive vesicles at the plasma membrane was significantly impaired. This correlated with deficient NADPH oxidase activation at the plasma membrane in response to Rab11 interference. Our data demonstrate that Munc13-4 is a Rab11-binding partner that regulates the final steps of Rab11-positive vesicle docking at the plasma membrane. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  11. Synthesis of O-[{sup 11}C]acetyl CoA, O-[{sup 11}C]acetyl-L-carnitine, and L-[{sup 11}C]carnitine labelled in specific positions, applied in PET studies on rhesus monkey

    Energy Technology Data Exchange (ETDEWEB)

    Jacobson, Gunilla B.; Watanabe, Yasuyoshi; Valind, Sven; Kuratsune, Hirohiko; Laangstroem, Bengt

    1997-07-01

    The syntheses of L-carnitine, O-acetyl CoA, and O-acetyl-L-carnitine labelled with {sup 11}C at the 1- or 2-position of the acetyl group or the N-methyl position of carnitine, using the enzymes acetyl CoA synthetase and carnitine acetyltransferase, are described. With a total synthesis time of 45 min, O-[1-{sup 11}C]acetyl CoA and O-[2-{sup 11}C]acetyl CoA was obtained in 60-70% decay-corrected radiochemical yield, and O-[1-{sup 11}C]acetyl-L-carnitine and O-[2-{sup 11}C]acetyl-L-carnitine in 70-80% yield, based on [1-{sup 11}C]acetate or [2-{sup 11}C]acetate, respectively. By an N-methylation reaction with [{sup 11}C]methyl iodide, L-[methyl-{sup 11}C]carnitine was obtained within 30 min, and O-acetyl-L-[methyl-{sup 11}C]carnitine within 40 min, giving a decay-corrected radiochemical yield of 60% and 40-50%, respectively, based on [{sup 11}C]methyl iodide. Initial data of the kinetics of the different {sup 11}C-labelled L-carnitine and acetyl-L-carnitines in renal cortex of anaesthetized monkey (Macaca mulatta) are presented.

  12. Understanding thermostability and pH dependent properties of proteins

    DEFF Research Database (Denmark)

    Galberg, Pernille

    The work performed in this thesis is part of a larger project (“Computational design of stable enzymes”) involving several research teams, which aimed to improve PROPKA (http://propka.ki.ku.dk) and to provide the scientific community with a computational protocol and associated PROPKA program......, which could be used for predicting mutations with expectation of increased thermostability at a certain pH value or a shifted pH activity optimum. The ability of a Bacillus circulans xylanase (BCX) mutant (N35D/A115E) to induce a decrease in pH activity optimum was evaluated by a pH dependent xylanase...

  13. A Family-Based Association Study of CYP11A1 and CYP11B1 Gene Polymorphisms With Autism in Chinese Trios.

    Science.gov (United States)

    Deng, Hong-Zhu; You, Cong; Xing, Yu; Chen, Kai-Yun; Zou, Xiao-Bing

    2016-05-01

    Autism spectrum disorder is a group of neurodevelopmental disorders with the higher prevalence in males. Our previous studies have indicated lower progesterone levels in the children with autism spectrum disorder, suggesting involvement of the cytochrome P-450scc gene (CYP11A1) and cytochrome P-45011beta gene (CYP11B1) as candidate genes in autism spectrum disorder. The aim of this study was to investigate the family-based genetic association between single-nucleotide polymorphisms, rs2279357 in the CYP11A1 gene and rs4534 and rs4541 in the CYP11B1 gene and autism spectrum disorder in Chinese children, which were selected according to the location in the coding region and 5' and 3' regions and minor allele frequencies of greater than 0.05 in the Chinese populations. The transmission disequilibrium test and case-control association analyses were performed in 100 Chinese Han autism spectrum disorder family trios. The genotype and allele frequency of the 3 single-nucleotide polymorphisms had no statistical difference between the children with autism spectrum disorder and their parents (P> .05). Transmission disequilibrium test analysis showed transmission disequilibrium of CYP11A1 gene rs2279357 single-nucleotide polymorphisms (χ(2)= 5.038,Pautism spectrum disorder exists within or near the CYP11A1 gene in the Han Chinese population. © The Author(s) 2015.

  14. A study of 11 Be an 11 Li halo nuclei by core breakup reactions

    International Nuclear Information System (INIS)

    Grevy, S.

    1997-01-01

    The study of light nuclei with large neutron excess are very useful for the understanding of nuclear matter far from stability. The nuclear halo phenomenon has been observed for the first time for Z 11 Be and 11 Li halo nuclei. In this channel, the neutron is supposed not to participate to the reaction and then, when detected, to carry out the same properties as in the halo nucleus. The deduced widths of the neutron momentum distributions are different from the one extracted from the core distributions and with the more recent theoretical models. From these studies, it is also stressed that the properties of the core are essential to understand the halo phenomenon. In particular, the correlation between the core vibrations and the halo neutron are able to explain the emergence of the halo in 11 Be. (author)

  15. Carbon-11 labelling of an inhibitor of acetylcholinesterase: [[sup 11]C]physostigmine

    Energy Technology Data Exchange (ETDEWEB)

    Bonnot-Lours, S.; Crouzel, C.; Prenant, C.; Hinnen, F. (CEA, 91 - Orsay (France). Service Hospitalier Frederic Joliot)

    1993-01-01

    Physostigmine, an alkaloid from calabar bean is a strong inhibitor of acetylcholinesterase and has been used clinically in the treatment of glaucoma, atropine intoxication, myasthenia gravis and more recently, in experimental trials in Alzheimer's disease. In order to study the AChE activity in the brain by positron emission tomography, we have undertaken the labelling of physostigmine with carbon-11. The synthesis involves the reaction of [[sup 11]C]methylisocyanate with eseroline. [[sup 11]C]Methylisocyanate was obtained by heating [[sup 11]C]acetylchloride with tetrabutylammonium azide in toluene. The synthesis of [[sup 11]C]CH[sub 3]COC1 involves the carbonation of methylmagnesium bromide in THF with cyclotron produced [[sup 11]C]carbon dioxide and the addition of phthaloyl dichloride. The [[sup 11]C]methylisocyanate is distilled into a solution of eseroline in ether with a small piece of sodium. After 10 minutes at 25[sup o]C, the solution is purified by HPLC and the appropriate fraction collected. Starting with 55.5 GBq (1.5 Ci) of [[sup 11]C]carbon dioxide, 0.92-1.48 GBq (25-40 mCi) of [[sup 11]C]Physostigmine are obtained 57 minutes after EOB. (author).

  16. EPICS release 3.11 specific documentation -- EPICS release notes for 3.11

    International Nuclear Information System (INIS)

    1994-01-01

    EPICS release 3.11 is now ready for user testing. A person who wants to set up a simplified application environment to boot an IOC and create databases using R3.11 should follow the directions in Appendix B, page 27, of the EPICS Source/Release Control Manual, Sept. 20, 1993. The R3.11 EPICS path at ANL/APS is /net/phebos/epics/R3.11 so the command to get the new release is /net/phebos/epics/R3.11/Unix/share/bin/getrel /net/phebos/epics/R3.11. An existing R3.8 short form report can be copied to this new directory and used to create a database. ANL/APS is currently testing an Application Developers Source/Release control system. It is not yet ready for general distribution. Attached are the EPICS R3.11 release notes

  17. Production of xylanase and CMCase on solid state fermentation in different residues by Thermoascus aurantiacus miehe Produção de xilanase e CMCase por fermentação em estado sólido em diferentes resíduos pelo fungo termofílico Thermoascus aurantiacus miehe

    Directory of Open Access Journals (Sweden)

    Roberto da Silva

    2005-09-01

    Full Text Available The use of waste as raw material is important for government economy and natural balance. The purpose of this work was to study the production of CMCase and xylanase by a Brazilian strain of Thermoascus aurantiacus in solid state fermentation (SSF using different agricultural residues (wheat bran, sugarcane bagasse, orange bagasse, corncob, green grass, dried grass, sawdust and corn straw as substrates without enrichment of the medium and characterize the crude enzymes.The study of the extracellular cellulolytic and hemicellulolytic enzymes showed that T. arantiacus is more xylanolytic than cellulolytic. The highest levels of enzymes were produced in corncob, grasses and corn straw. All the enzymes were stable at room temperature by 24 h over a broad pH range (3.0-9.0 and also were stable at 60ºC for 1 h. The optimum pH and temperature for xylanase and CMCase were 5.0-5.5 and 5.0 and 75ºC, respectively. The microorganism grew quickly in stationary, simple and low cost medium. The secreted extracellular enzymes presented properties that match with those frequently required in industrial environment.O emprego de residuos como matéria prima é importante como estrategia governamental e para o balanço ambiental. O propósito deste trabalho foi estudar a produção de CMCase e xilanase de uma linhagem de Thermoascus aurantiacus isolado de solo brasileiro em fermentação em estado sólido (SSF usando diferentes resíduos agrícolas (farelo de trigo, bagaço de cana, bagaço de laranja, sabugo de milho, grama verde, grama seca, serragem de eucalipto e palha de milho como substratos sem enriquecimento dos meios e caracterizar as enzimas. O estudo das enzimas hemiceluloliticas extracelulares mostrou que o fungo T. arantiacus é mais xilanolítico do que celulolítico. Ele produziu maiores níveis das enzimas em meios contendo sabugo de milho, grama e palha de milho. Todas as enzimas foram estáveis por 24 h à temperatura ambiente numa ampla faixa

  18. On space--time Killing tensors with a Segre characteristic [(11)(11)

    International Nuclear Information System (INIS)

    Hauser, I.; Malhiot, R.J.

    1976-01-01

    We consider the set of all space--times which admit a Killing tensor K/sub αβ/ whose Segre characteristic is [(11)(11)] and whose eigenvalues lambda 1 and lambda 2 satisfy the condition that dlambda 1 X dlambda 2 is not null. We prove that there locally exist scalar fields phi 3 , phi 4 such that lambda 1 , lambda 2 , phi 3 , phi 4 constitute a chart, and dlambda 1 x dlambda 2 = dlambda/sup i/ x d phi/sup r/ = 0 for i=1, 2 and r = 3, 4. Also, relative to this coordinate system, the metric has the same general form as Carter's Hamilton--Jacobi separable metric except that his arbitrary functions of lambda/sup i/ are replaced by functions of lambda/sup i/, phi 3 , phi 4 . If R/sup αβ/(del/sub α/lambda 1 )(del/sub β/lambda 2 ) = 0, there exists a two-parameter Abelian group of isometries which commute with K/sub αβ/; also /phi 3 ,phi 4 ) can be chosen so that par.delta par.delta phi 3 and par.delta/par.delta phi 4 are the Killing vectors. R/sup αβ/(del/sub α/lambda 1 )(del sub β/lambda 2 ) = 0 is necessary and sufficient for Schroedinger equation separability in case (a) of Carter. A Newtonian analog of our results is discussed

  19. Evaluation of (+)-p-[{sup 11}C]methylvesamicol for mapping sigma{sub 1} receptors: a comparison with [{sup 11}C]SA4503

    Energy Technology Data Exchange (ETDEWEB)

    Ishiwata, Kiichi [Positron Medical Center, Tokyo Metropolitan Institute of Gerontology, Tokyo 173-0022 (Japan)]. E-mail: ishiwata@pet.tmig.or.jp; Kawamura, Kazunori [Positron Medical Center, Tokyo Metropolitan Institute of Gerontology, Tokyo 173-0022 (Japan); SHI Accelerator Service Ltd., Tokyo 141-0032 (Japan); Yajima, Kazuyoshi [The Medical and Pharmacological Research Center Foundation, Hakui 920-0631 (Japan); QingGeLeTu [Positron Medical Center, Tokyo Metropolitan Institute of Gerontology, Tokyo 173-0022 (Japan); Mori, Hirofumi [Advanced Science Research Center, Kanazawa University, Kanazawa 920-8640 (Japan); Shiba, Kazuhiro [Advanced Science Research Center, Kanazawa University, Kanazawa 920-8640 (Japan)

    2006-05-15

    Vesamicol is a leading compound for positron emission tomography (PET) and single photon emission computed tomography (SPECT) tracers for mapping the vesicular acetylcholine transporter (VAChT). Recently, we found that (+)-p-methylvesamicol ((+)-PMV) has low affinity for VAChT (K {sub i}=199 nM), but has moderate to high affinity for sigma receptors: K {sub i}=3.0 nM for sigma{sub 1} and K {sub i}=40.7 nM for sigma{sub 2}, and that sigma{sub 1}-selective SA4503 (K {sub i}=4.4 nM for sigma{sub 1} and K {sub i}=242 nM for sigma{sub 2}) has moderate affinity for VAChT (K {sub i}=50.2 nM). In the present study, we examined the potential of (+)-[{sup 11}C]PMV as a PET radioligand for mapping sigma{sub 1} receptors as compared with [{sup 11}C]SA4503. In rat brain, similar regional distribution patterns of (+)-[{sup 11}C]PMV and [{sup 11}C]SA4503 were shown by tissue dissection and by ex vivo autoradiography. Blocking experiments using ({+-})-PMV (-)-vesamicol, SA4503, haloperidol and ({+-})-pentazocine showed that the two tracers specifically bound to sigma{sub 1} receptors, and that [{sup 11}C]SA4503 exhibited greater specific binding than (+)-[{sup 11}C]PMV. No sign of VAChT-specific binding by [{sup 11}C]SA4503 was observed in the striatum, which is rich in VAChT sites. In conclusion, (+)-[{sup 11}C]PMV specifically bound to sigma{sub 1} receptors in the brain, but to a lesser extent than [{sup 11}C]SA4503, suggesting that (+)-[{sup 11}C]PMV is a less preferable PET ligand than [{sup 11}C]SA4503. On the other hand, the moderate affinity of [{sup 11}C]SA4503 for VAChT is negligible in vivo.

  20. The Loss of Vacuolar Protein Sorting 11 (vps11) Causes Retinal Pathogenesis in a Vertebrate Model of Syndromic Albinism

    Science.gov (United States)

    Thomas, Jennifer L.; Vihtelic, Thomas S.; denDekker, Aaron D.; Willer, Gregory; Luo, Xixia; Murphy, Taylor R.; Gregg, Ronald G.; Hyde, David R.

    2011-01-01

    Purpose. To establish the zebrafish platinum mutant as a model for studying vision defects caused by syndromic albinism diseases such as Chediak-Higashi syndrome, Griscelli syndrome, and Hermansky-Pudlak syndrome (HPS). Methods. Bulked segregant analysis and candidate gene sequencing revealed that the zebrafish platinum mutation is a single-nucleotide insertion in the vps11 (vacuolar protein sorting 11) gene. Expression of vps11 was determined by RT-PCR and in situ hybridization. Mutants were analyzed for pigmentation defects and retinal disease by histology, immunohistochemistry, and transmission electron microscopy. Results. Phenocopy and rescue experiments determined that a loss of Vps11 results in the platinum phenotype. Expression of vps11 appeared ubiquitous during zebrafish development, with stronger expression in the developing retina and retinal pigmented epithelium (RPE). Zebrafish platinum mutants exhibited reduced pigmentation in the body and RPE; however, melanophore development, migration, and dispersion occurred normally. RPE, photoreceptors, and inner retinal neurons formed normally in zebrafish platinum mutants. However, a gradual loss of RPE, an absence of mature melanosomes, and the subsequent degradation of RPE/photoreceptor interdigitation was observed. Conclusions. These data show that Vps11 is not necessary for normal retinal development or initiation of melanin biosynthesis, but is essential for melanosome maturation and healthy maintenance of the RPE and photoreceptors. PMID:21330665

  1. Synthesis of [(11)C]Am80 via Novel Pd(0)-Mediated Rapid [(11)C]Carbonylation Using Arylboronate and [(11)C]Carbon Monoxide.

    Science.gov (United States)

    Takashima-Hirano, Misato; Ishii, Hideki; Suzuki, Masaaki

    2012-10-11

    (11)C-labeled methylbenzoates [(11)C]4a-d were synthesized using Pd(0)-mediated rapid cross-coupling reactions employing [(11)C]carbon monoxide and arylboronic acid neopentyl glycol esters 3a-d under atmospheric pressure in methanol-dimethylformamide (MeOH-DMF), in radiochemical yields of 12 ± 5-26 ± 13% (decay-corrected based on [(11)C]O). The reaction conditions were highly favorable for the synthesis of [(11)C]Am80 ([(11)C]2) and [(11)C]methyl 4-((5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)carbamoyl)benzoate ([(11)C]2-Me) using 4-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-N-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)benzamide (5), both of which produced a decay-corrected radiochemical yield (RCY) of 26 ± 13%, with >99% radiochemical purity and an average specific radioactivity of 44 GBq/μmol. The yields of [(11)C]4a, [(11)C]2-Me, and [(11)C]2 were improved by the use of a 2-fold excess of the solvents and reagents under the same conditions to give respective yields of 66 ± 8, 65 ± 7, and 48 ± 2%.

  2. The synthesis of no-carrier-added [11C]urea from [11C]carbon dioxide and application to [11C]uracil synthesis

    International Nuclear Information System (INIS)

    Chakraborty, P.K.; Mangner, T.J.; Chugani, H.T.

    1997-01-01

    No-carrier-added [ 11 C]urea has been synthesized by bubbling cyclotron-produced [ 11 C]-carbon dioxide directly into a tetrahydrofuran solution of lithium bis(trimethylsilyl)amide, followed by hydrolysis of the C-11 labeled adduct with aqueous ammonium chloride. Using this simple, one-pot method, [ 11 C]urea was produced in 55-70% radiochemical yield (decay-corrected) in 16 min following end-of-bombardment (or in 10 min following the introduction of the [ 11 C]carbon dioxide. The [ 11 C]urea thus produced was converted to [ 11 C]uracil (carrier-added) in 40-75% decay-corrected radiochemical yield by condensation with diethyl malate in presence of fuming sulfuric acid. (author)

  3. Cellobiose dehydrogenase entrapped within specifically designed Os-complex modified electrodeposition polymers as potential anodes for biofuel cells

    International Nuclear Information System (INIS)

    Shao, Minling; Guschin, Dmitrii A.; Kawah, Zahma; Beyl, Yvonne; Stoica, Leonard; Ludwig, Roland; Schuhmann, Wolfgang; Chen, Xingxing

    2014-01-01

    Electron-transfer pathways between cellobiose dehydrogenase from Myriococcum thermophilum (MtCDH) and the related flavodehydrogenase domain (FAD-MtCDH) and electrodes were evaluated using specifically designed Os-complex modified electrodeposition paints (EDPs). The properties of the Os-complex modified EDPs were varied by variation of the monomer composition, the coordination sphere of the polymer-bound Os-complexes, and the length and flexibility of the spacer chain between Os complex and polymer backbone. The MtCDH-to-EDP weight ratio, the pH value, as well as the operational temperature have been optimized

  4. Examining the Potential of Plasma-Assisted Pretreated Wheat Straw for Enzyme Production by Trichoderma reesei

    DEFF Research Database (Denmark)

    Rodríguez Gómez, Divanery; Lehmann, Linda Olkjær; Schultz-Jensen, Nadja

    2012-01-01

    Plasma-assisted pretreated wheat straw was investigated for cellulase and xylanase production by Trichoderma reesei fermentation. Fermentations were conducted with media containing washed and unwashed plasma-assisted pretreated wheat straw as carbon source which was sterilized by autoclavation....... To account for any effects of autoclavation, a comparison was made with unsterilized media containing antibiotics. It was found that unsterilized washed plasma-assisted pretreated wheat straw (which contained antibiotics) was best suited for the production of xylanases (110 IU ml(-1)) and cellulases (0...... other nonrefined feedstocks suggests that plasma pretreated wheat straw is a promising and suitable substrate for cellulase and hemicellulase production....

  5. Standardy 802.11n a 802.11e v bezdrátových sítích

    OpenAIRE

    Novák, Ondřej

    2009-01-01

    Tato bakalářská práce je zaměřena na problematiku standardů bezdrátových sítí. Teoreticky popisuje nejznámější standardy skupiny standardů IEEE 802.11, jejich funkce a chování. Dále práce podrobněji rozebírá nový standard 802.11n a specifikaci standardu 802.11e, WMM. Práce obsahuje zprávu o praktických testech standardů 802.11n a WMM, které slouží pro objektivní posouzení schopností těchto standardů. This bachelor thesis is focused on broad issue of wireless network standards. It theoretic...

  6. Selection and characterisation of a xylitol-derepressed Aspergillus niger mutant that is apparently impaired in xylitol transport

    NARCIS (Netherlands)

    Vondervoort, van de P.J.I.; Groot, de M.J.L.; Ruijter, G.J.G.; Visser, J.

    2006-01-01

    Aspergillus niger is known for its biotechnological applications, such as the use of xylanase enzyme for the degradation of hemicellulose. Depending on culture conditions, several polyols may also be accumulated, such as xylitol during D-xylose oxidation. Also during industrial fermentation of

  7. Studies on the enzymes produced by Basidiomycetes. Part 1. The production of crude enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Hong, J. S.; Kim, D.H.

    1981-01-01

    Cellulase, protease, and xylanase, formation by the basidiomycetes, Pleurotus ostreatus 301 and Lentinus edodes 3-1 in growth on rice straw medium were studied. Cultural conditions adequate for enzyme production and effects of various materials and inorganic salts added to the rice straw media were investigated. Lentinus edodes 3-1 was an excellent producer of cellulase and xylanase, and Pleurotus ostreatus 301 of protease. The optimum conditions for enzyme production were 30 degrees for cellulase production and at 25 degrees for xylanase and protease production, with 75% moisture content and initial pH of 5.0-6.0. The appropriate incubation times for enzyme production were 30 days and 35 days for Pleurotus ostreatus 301 and Lentinus edodes 3-1, respectively. Among the various materials added, defatted soybean, defatted rape seed, or defatted sesame were all effective in enzyme production but reduced mycelial growth. Rice bran was also effective, particularly at a 30% concentration. The addition of inorganic salts enhanced enzyme production. Among inorganic salts, the optimum concentration of CaCO3 was 5%, and that of CaSO4 was 2%.

  8. A 1.37-Mb 12p11.22-p11.21 deletion coincident with a 367-kb 22q11.2 duplication detected by array comparative genomic hybridization in an adolescent girl with autism and difficulty in self-care of menstruation.

    Science.gov (United States)

    Chen, Chih-Ping; Lin, Shuan-Pei; Chern, Schu-Rern; Wu, Peih-Shan; Su, Jun-Wei; Lee, Chen-Chi; Wang, Wayseen

    2014-03-01

    To present an array comparative genomic hybridization (aCGH) characterization of a 12p11.22-p11.21 microdeletion and 22q11.2 microduplication in an adolescent girl with autism, mental retardation, facial dysmorphism, microcephaly, behavior problems, and an apparently balanced reciprocal translocation of t(8;12)(q24.3;p11.2). A 13-year-old girl was referred to the hospital because of autism, mental retardation, and difficulty in the self-care of her menstruation. Cytogenetic analysis revealed an apparently balanced reciprocal translocation and a karyotype of 46,XX,t(8;12) (q24.3;p11.2)dn. The girl manifested microcephaly, hypertelorism, flat facial profile, prominent forehead, thick scalp hair, upslanting palpebral fissures, broad nasal bridge, bulbous nose, right simian crease, bilateral clinodactyly of the fifth fingers, bilateral pes cavus, learning difficulties, mental retardation, emotional instability, cognitive impairment, behavior problems, jumping-like gaits, and autistic spectrum disorder. aCGH was performed to evaluate genomic imbalance in this patient. aCGH analysis revealed a 1.37-Mb 12p11.22-p11.21 microdeletion or arr [hg 19] 12p11.22-p11.21 (30,645,008-32,014,774)×1 and a 367-kb 22q11.21 microduplication or arr [hg 19] 22q11.21 (18,657,470-19,024,306)×3. The 1.37-Mb 12p11.22-p11.21 microdeletion encompassed 26 genes including IPO8, CAPRIN2, and DDX11, and the 367-kb 22q11.21 microduplication encompassed 20 genes including USP18, DGCR6, PRODH, and DGCR2. An apparently balanced translocation may be in fact affected by concurrent deletion and duplication in two different chromosomal regions. Our presentation provides information on diagnostic phenotype of 12p11.22-p11.21 microdeletion and 22q11.2 microduplication. Copyright © 2014. Published by Elsevier B.V.

  9. A novel approach for a C-11C bond formation: synthesis of 17α-([11C]prop-1-ynyl)-3-methoxy-3,17β-estradiol

    International Nuclear Information System (INIS)

    Wuest, F.; Zessin, J.

    2002-01-01

    A novel method for a 11 C-C bond formation was developed, employing a cross-coupling reaction between a terminal acetylene and [ 11 C]methyl iodide. The method was used for the synthesis of 17α-([ 11 C]prop-1-ynyl)-3-methoxy-3,17β-estadiol. (orig.)

  10. Synthesis of electrophilic 11C-synthons: 11C-cyanogen bromide and some 11C-cyano reagents

    International Nuclear Information System (INIS)

    Westerberg, G.; Malmborg, P.; Langstroem, B.

    1990-01-01

    A fast, simple, and reliable method has been developed for the synthesis of carbon-11 labelled cyanogen bromide and several of its stabilized amine salts or cyanates. Preparation of cyanogen bromide is by bromination (bromine gas) of carbon-11 labelled hydrocyanic acid, followed by reaction with a tertiary amine or a phenol to yield the ammonium salts or cyanates

  11. Routine production of H11CN and :11C:-1-aminocyclopentanecarboxylic acid

    International Nuclear Information System (INIS)

    Sambre, J.; Vandecasteele, C.; Geothals, P.; Rabi, N.A.; Haver, D. van; Slegers, G.

    1985-01-01

    The production of H 11 CN using the 14 N(p,α) 11 C reaction, was studied. The yield is 140 mCi/μA at saturation (EOB). From 1.8 Ci of H 11 CN 140 mCi of [ 11 C]ACPC are produced routinely and under remote-control. Chemical and pharmaceutical controls showed that the product is suitable for injection. (author)

  12. Purification, Characterization, and Optimum Conditions of Fermencin SD11, a Bacteriocin Produced by Human Orally Lactobacillus fermentum SD11.

    Science.gov (United States)

    Wannun, Phirawat; Piwat, Supatcharin; Teanpaisan, Rawee

    2016-06-01

    Fermencin SD11, a bacteriocin produced by human orally Lactobacillus fermentum SD11, was purified, characterized, and optimized in conditions for bacterial growth and bacteriocin production. Fermencin SD11 was purified using three steps of ammonium sulfate precipitation, gel filtration chromatography, and reverse-phase high-performance liquid chromatography. The molecular weight was found to be 33,000 Da using SDS-PAGE and confirmed as 33,593.4 Da by liquid chromatography-mass spectrometry. Fermencin SD11 exhibited activity against a wide range of oral pathogens including cariogenic and periodontogenic pathogens and Candida. The active activity was stable between 60 - 80 °C in a pH range of 3.0 to 7.0. It was sensitive to proteolytic enzymes (proteinase K and trypsin), but it was not affected by α-amylase, catalase, lysozyme, and saliva. The optimum conditions for growth and bacteriocin production of L. fermentum SD11 were cultured at acidic with pH of 5.0-6.0 at 37 or 40 °C under aerobic or anaerobic conditions for 12 h. It is promising that L. fermentum SD11 and its bacteriocin may be an alternative approach for promoting oral health or prevention of oral diseases, e.g., dental caries and periodontitis, which would require further clinical trials.

  13. The determination of unchanged [11C]diprenorphine, [11C]L-deprenyl and [11C]raclopride in plasma by HPLC

    International Nuclear Information System (INIS)

    Luthra, S.K.; Turton, D.R.; Price, G.; Ahier, R.; Martin, F.; Hume, S.; Cremer, J.

    1990-01-01

    In PET studies a knowledge of the fraction of radioactivity in plasma representing unchanged 11 C-labelled ligand/tracer is essential for correction of the arterial plasma input curve. A number of different approaches (e.g. HPLC and TLC/OPTLC) to the determination of unchanged radio-ligand/tracer have been reported for 11 C-labelled carfentanil, raclopride and L-deprenyl and for 76 Br-labelled bromolisuride. The authors report here how they have applied the simple work-up and analytical procedure, originally reported for [ 11 C]carfentanil, to establish the percentage of unmchanged 11 C-labelled diprenorphine, L-deprenyl and raclopride in human and animal plasma as a function of time

  14. Development of [11C]/[3H]THK-5351 – A potential novel carbon-11 tau imaging PET radioligand

    International Nuclear Information System (INIS)

    Stepanov, Vladimir; Svedberg, Marie; Jia, Zhisheng; Krasikova, Raisa; Lemoine, Laetitia; Okamura, Nobujuki; Furumoto, Shozo; Mitsios, Nicholas; Mulder, Jan; Långström, Bengt; Nordberg, Agneta; Halldin, Christer

    2017-01-01

    Introduction: Due to the rise in the number of patients with dementia the imperative for finding new diagnostic and treatment options becomes ever more pressing. While significant progress has been made in PET imaging of Aβ aggregates both in vitro and in vivo, options for imaging tau protein aggregates selectively are still limited. Based on the work previously published by researchers from the Tohoku University, Japan, that resulted in the development of [ 18 F]THK-5351, we have undertaken an effort to develop a carbon-11 version of the identical structure - [ 11 C]THK-5351. In parallel, THK-5351 was also labeled with tritium ([ 3 H]THK-5351) for use in in vitro autoradiography (ARG). Methods: The carbon-11 labeling was performed starting with di-protected enantiomeric pure precursor - tert-butyl 5-(6-((2S)-3-fluoro-2-(tetrahydro-2H-pyran-2-yloxy)propoxy)quinolin-2-yl) pyridin-2-yl carbamate, which was reacted with [ 11 C]MeI, using DMF as the solvent and NaH as base, followed by deprotection with trifluoroacetic acid/water mixture, resulting in enantiomerically pure carbon-11 radioligand, [ 11 C]THK-5351 - (S)-1-fluoro-3-(2-(6-([ 11 C]methylamino)pyridin-3-yl)quinolin-6-yloxy) propan-2-ol. Tritium labeling and purification of [ 3 H]THK-5351 were undertaken using similar approach, resulting in [ 3 H]THK-5351 with RCP >99.8% and specific radioactivity of 1.3 GBq/μmol. Results: [ 11 C]THK-5351 was produced in good yield (1900 ± 355 MBq), specific radioactivity (SRA) (361 ± 119 GBq/μmol at EOS + 20 min) and radiochemical purity (RCP) (>99.8%), with enantiomeric purity of 98.7%. [ 3 H]THK-5351 was evaluated for ARG of tau binding in post-mortem human brain tissue using cortical sections from one AD patient and one control subject. [ 3 H]THK-5351 binding density was higher in the AD patient compared to the control subject, the binding was displaced by unlabeled THK-5351 confirming specific [ 3 H]THK-5351 binding.

  15. Interleukin-11 Receptor Is a Candidate Target for Ligand-Directed Therapy in Lung Cancer: Analysis of Clinical Samples and BMTP-11 Preclinical Activity.

    Science.gov (United States)

    Cardó-Vila, Marina; Marchiò, Serena; Sato, Masanori; Staquicini, Fernanda I; Smith, Tracey L; Bronk, Julianna K; Yin, Guosheng; Zurita, Amado J; Sun, Menghong; Behrens, Carmen; Sidman, Richard L; Lee, J Jack; Hong, Waun K; Wistuba, Ignacio I; Arap, Wadih; Pasqualini, Renata

    2016-08-01

    We previously isolated an IL-11-mimic motif (CGRRAGGSC) that binds to IL-11 receptor (IL-11R) in vitro and accumulates in IL-11R-expressing tumors in vivo. This synthetic peptide ligand was used as a tumor-targeting moiety in the rational design of BMTP-11, which is a drug candidate in clinical trials. Here, we investigated the specificity and accessibility of IL-11R as a target and the efficacy of BMTP-11 as a ligand-targeted drug in lung cancer. We observed high IL-11R expression levels in a large cohort of patients (n = 368). In matching surgical specimens (i.e., paired tumors and nonmalignant tissues), the cytoplasmic levels of IL-11R in tumor areas were significantly higher than in nonmalignant tissues (n = 36; P = 0.003). Notably, marked overexpression of IL-11R was observed in both tumor epithelial and vascular endothelial cell membranes (n = 301; P < 0.0001). BMTP-11 induced in vitro cell death in a representative panel of human lung cancer cell lines. BMTP-11 treatment attenuated the growth of subcutaneous xenografts and reduced the number of pulmonary tumors after tail vein injection of human lung cancer cells in mice. Our findings validate BMTP-11 as a pharmacologic candidate drug in preclinical models of lung cancer and patient-derived tumors. Moreover, the high expression level in patients with non-small cell lung cancer is a promising feature for potential translational applications. Copyright © 2016 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

  16. Neuronal RING finger protein 11 (RNF11 regulates canonical NF-κB signaling

    Directory of Open Access Journals (Sweden)

    Pranski Elaine L

    2012-04-01

    Full Text Available Abstract Background The RING domain-containing protein RING finger protein 11 (RNF11 is a member of the A20 ubiquitin-editing protein complex and modulates peripheral NF-κB signaling. RNF11 is robustly expressed in neurons and colocalizes with a population of α-synuclein-positive Lewy bodies and neurites in Parkinson disease patients. The NF-κB pathway has an important role in the vertebrate nervous system, where the absence of NF-κB activity during development can result in learning and memory deficits, whereas chronic NF-κB activation is associated with persistent neuroinflammation. We examined the functional role of RNF11 with respect to canonical NF-κB signaling in neurons to gain understanding of the tight association of inflammatory pathways, including NF-κB, with the pathogenesis of neurodegenerative diseases. Methods and results Luciferase assays were employed to assess NF-κB activity under targeted short hairpin RNA (shRNA knockdown of RNF11 in human neuroblastoma cells and murine primary neurons, which suggested that RNF11 acts as a negative regulator of canonical neuronal NF-κB signaling. These results were further supported by analyses of p65 translocation to the nucleus following depletion of RNF11. Coimmunoprecipitation experiments indicated that RNF11 associates with members of the A20 ubiquitin-editing protein complex in neurons. Site-directed mutagenesis of the myristoylation domain, which is necessary for endosomal targeting of RNF11, altered the impact of RNF11 on NF-κB signaling and abrogated RNF11’s association with the A20 ubiquitin-editing protein complex. A partial effect on canonical NF-κB signaling and an association with the A20 ubiquitin-editing protein complex was observed with mutagenesis of the PPxY motif, a proline-rich region involved in Nedd4-like protein interactions. Last, shRNA-mediated reduction of RNF11 in neurons and neuronal cell lines elevated levels of monocyte chemoattractant protein 1 and

  17. Observation of Cleft Palate in an Individual with SOX11 Mutation: Indication of a Role for SOX11 in Human Palatogenesis.

    Science.gov (United States)

    Khan, Umair; Study, Ddd; Baker, Eleanor; Clayton-Smith, Jill

    2018-03-01

    Point mutations and deletions within the SOX11 gene have recently been described in individuals with a rare variant of Coffin-Siris syndrome, OMIM 615866, an intellectual disability syndrome with associated features of nail hypoplasia, microcephaly, and characteristic facial features including a wide mouth and prominent lips. We describe a further patient with a mutation in SOX11 and phenotype resembling mild Coffin-Siris syndrome. This boy had a cleft palate, a feature not previously seen in other patients with SOX11 mutations. We discuss This adds to the current evidence that SOX11 is a gene involved in palatogenesis.

  18. Loss of heterozygosity at 11p13 and 11p15 in Wilms tumor: a study of 22 cases from India.

    Science.gov (United States)

    Sigamani, Elanthenral; Wari, Mohammad Nahidul; Iyer, Venkateswaran K; Agarwala, Sandeep; Sharma, Arundhati; Bakhshi, Sameer; Dinda, Amit

    2013-03-01

    11p13 and 11p15 loss of heterozygosity (LOH) in Wilms tumor (WT), the commonest molecular pathogenetic event in WT, shows variation in different parts of the world. The present study looked for the presence of 11p13 and 11p15 LOH as well as nephrogenic rests in WT occurring in India. Twenty-two cases of WT were subjected to thorough pathological examination for presence of nephrogenic rests. Fresh frozen tissue was evaluated for LOH at 11p13 and 11p15, using PCR for microsatellite markers. Among twenty-two consecutive cases of WT, 20 were unilateral and 2 were bilateral. 6/22 showed LOH at 11p13 (27.7 %) and 1/22 showed LOH at 11p15 (4.54 %). 2/22 cases showed presence of nephrogenic rests. One of the cases with LOH at 11p13 had intralobar nephrogenic rest in the adjacent kidney. One specimen had perilobar nephrogenic rest in the adjacent kidney but did not show LOH for either 11p13 or 11p15 in the tumor. LOH at 11p13 is seen in 27.27 % of WT in India, which is similar to reports in the English language literature. LOH at 11p15 was seen in 4.54 % of WT, which is lower than that reported from Western subjects.

  19. ISA100.11a*: The ISA100.11a extension for supporting energy-harvested I/O devices

    NARCIS (Netherlands)

    Zand, P.; Mathews, E.; Das, Kallol; Dilo, Arta; Havinga, Paul J.M.

    Wireless standards developed for industrial applications such as ISA100.11a and WirelessHART, generally use centralized management approaches. However, such centralized approaches cannot cope with network dynamicity in real-time manner. They also incur high management overhead and latency.

  20. A FORTRAN-compatible program package for the control of CAMAC-systems by a PDP-11 (CA11-A/DEC, Type 1533A/BORER)

    International Nuclear Information System (INIS)

    Lengauer, C.

    1975-01-01

    The described software serves for the control of CAMAC-systems by a PDP-11 Computer with one DEC CA11-A Branch-Driver, respectively up to ten BORER Type 1533A Single-Crate-Controllers under the Operating System DOS V08. The software consists of three parts: 1) a subroutine library for programming in FORTRAN, 2) a macro library for programming in Assembler (for time-critical problems), 3) a loadable CAMAC-Driver for controlling the system by input of single CAMAC-commands at the terminal. Programs which apply the first two parts can be written independently of the CAMAC-Controller used at runtime. (orig.) [de

  1. Radiosynthesis of [{sup 11}C]D.P.A.-713, [{sup 11}C]D.P.A.-715 and [{sup 11}C]clinme, selected carbon-11-labelled novel potential radioligands for imaging the peripheral benzodiazepine receptors with PET

    Energy Technology Data Exchange (ETDEWEB)

    Dolle, F.; Thominiaux, C.; Hinnen, F.; Demphel, S.; Le helleix, S.; Chauveau, F.; Boutin, H.; Herard, A.S.; Hantraye, P.; Tavitian, B. [Service Hospitalier Frederic Joliot, I2BM/DSV, 91 - Orsay (France); Kassiou, M.; James, M.; Creelman, A.; Fulton, R. [Sydney Univ., Brain and Mind Research Institute, NSW (Australia); Kassiou, M. [Sydney Univ., Discipline of Medical Radiations, Sciences and School of Chemistry, NSW (Australia); Katsifis, A.; Greguric, I.; Mattner, F.; Loch, C. [Radiopharmaceuticals Research Institute, ANSTO, NSW (Australia); Selleri, S. [Degli Studi di Firenze Univ., Dipt. di Scienze Farmaceutiche (Italy)

    2008-02-15

    {sup 11}C P.K.11195 is not only the oldest, but also the most widely used PET radiotracer for in vivo imaging of the peripheral benzodiazepine receptors (P.B.R. or translocator protein (18 kDa, T.S.P.O.). With the aim of developing a new PET imaging probe for the in vivo study of the P.B.R., two pyrazol [1,5-a]pyrimidineacetamides (D.P.A.-713 and D.P.A.-715) and one imidazol[1,2-a]pyridine-acetamide (C.L.I.N.M.E.) were radiolabelled with the positron emitters carbon{sup 11} (half life: 20.38 min) [1-5]. Briefly, C.L.I.N.M.E. (2-[6-chloro-2(4-iodophenyl)-imidazol[1,2-a]pyridin-3-yl] -N-ethyl-N-methyl-acetamide) was labelled at its methyl-acetamide moity chain from the corresponding nor-analogue using[{sup 11}C]methyl iodide (in D.M.S.O./D.M.F (100/200 {mu}L) containing powdered K.O.H. (3-5 mg) at 110 degrees C for 3 min. D.P.A.-713 (N,N-diethyl-2-[2-(4-methoxy-phenyl)-5,7-dimethyl-pyrazolo[1,5-a]pyrimidin -3-yl]acetamide) and D.P.A.-715 (N,N-diethyl-2-[2-(4-methoxy-phenyl)-5,7-bis-tri-fluoro-methyl-pyrazolo [1,5-a]pyrimidin-3-yl]acetamide) were labelled at their aromatic methoxy groups from the corresponding nor-derivatives using [{sup 11}C]methyl triflate (in acetone (300{mu}L) containing aq. 3 M NaOH (4{mu}L) at 110 degrees C for 1 min). All radioligands were purified using semi preparative Zorbax reverse phase H.P.L.C., were adequately formulated for in vivo injection within 30 min and were found to be > 95% chemically and radiochemically pure. (N.C.)

  2. 41 CFR 301-11.11 - How do I make my lodging reservations?

    Science.gov (United States)

    2010-07-01

    ... your agency travel management service as required by part 301-50 of this chapter. [FTR Amdt. 108, 67 FR... reservations? 301-11.11 Section 301-11.11 Public Contracts and Property Management Federal Travel Regulation System TEMPORARY DUTY (TDY) TRAVEL ALLOWANCES ALLOWABLE TRAVEL EXPENSES 11-PER DIEM EXPENSES General...

  3. Producción y purificación parcial de enzimas hidrolíticas de Aspergillus ficuum en fermentación sólida sobre residuos agroindustriales

    Directory of Open Access Journals (Sweden)

    Marcia Costa

    2010-07-01

    Full Text Available Título en inglés: Production and partial purification of Aspergillus ficuum hydrolytic enzymes in solid state fermentation of agroindustrial residues Resumen En el presente trabajo se describe la producción de las enzimas fitasa, celulasa, xilanasa y proteasa con Aspergillus ficuum cepa DSM 932 mediante fermentación en estado sólido (SSF usando torta de canola y pomaza de cranberry como sustratos. Como medida indirecta de la producción de las enzimas se usó en cada caso la actividad enzimática. la torta de canola resultó ser un mejor sustrato para fitasa, celulasa y xilanasa, en tanto que la pomaza de cranberry resultó ser un sustrato potencial para proteasa. Mediante ultrafiltración escalonada fue posible purificar parcialmente los extractos enzimáticos de fitasa, celulasas y xilanasas, obtenidos a partir de torta de canola. La fitasa resultó tener un tamaño >100 kDa, en tanto que las celulasas y xilanasas presentan actividad en los retenidos de 10, 30 y 50 kDa, lo que indicaría que las isoenzimas de ambos complejos tienen pesos moleculares que oscilan entre 10 y 100 kDa. Palabras clave: fermentación sólida; torta de canola; Aspergillus ficuum; fitasa; xilanasa. Abstract In this paper, describes the production of the enzymes phytase, cellulase, xylanase and protease by Aspergillus ficuum DSM 932 strain, in solid state fermentation (SSF using canola cake and cranberry pomace as substrates. The enzyme activity was used in each case as an indirect measure of the enzymes production. Canola meal turned out to be a better substrate for phytase, cellulase and xylanase, while cranberry pomace was found to be a potential substrate for protease. Various ultrafiltration operations were carried out, decreasing the cut off membranes out in order to purify partially extracts of enzymes phytase, cellulase and xylanase, obtained from canola meal. Phytase was found to have a size >100 kDa, whereas cellulase and xylanase activity present in the

  4. PET imaging of focal demyelination and remyelination in a rat model of multiple sclerosis: comparison of [{sup 11}C]MeDAS, [{sup 11}C]CIC and [{sup 11}C]PIB

    Energy Technology Data Exchange (ETDEWEB)

    Paula Faria, Daniele de [University of Groningen, University Medical Center Groningen, Department of Nuclear Medicine and Molecular Imaging, Groningen (Netherlands); University of Groningen, University Medical Center Groningen, Department of Neuroscience, Groningen (Netherlands); University of Sao Paulo Medical School, Center of Nuclear Medicine, Sao Paulo (Brazil); Copray, Sjef [University of Groningen, University Medical Center Groningen, Department of Neuroscience, Groningen (Netherlands); Sijbesma, Jurgen W.A.; Willemsen, Antoon T.M.; Dierckx, Rudi A.J.O.; Vries, Erik F.J. de [University of Groningen, University Medical Center Groningen, Department of Nuclear Medicine and Molecular Imaging, Groningen (Netherlands); Buchpiguel, Carlos A. [University of Sao Paulo Medical School, Center of Nuclear Medicine, Sao Paulo (Brazil)

    2014-05-15

    In this study, we compared the ability of [{sup 11}C]CIC, [{sup 11}C]MeDAS and [{sup 11}C]PIB to reveal temporal changes in myelin content in focal lesions in the lysolecithin rat model of multiple sclerosis. Pharmacokinetic modelling was performed to determine the best method to quantify tracer uptake. Sprague-Dawley rats were stereotactically injected with either 1 % lysolecithin or saline into the corpus callosum and striatum of the right brain hemisphere. Dynamic PET imaging with simultaneous arterial blood sampling was performed 7 days after saline injection (control group), 7 days after lysolecithin injection (demyelination group) and 4 weeks after lysolecithin injection (remyelination group). The kinetics of [{sup 11}C]CIC, [{sup 11}C]MeDAS and [{sup 11}C]PIB was best fitted by Logan graphical analysis, suggesting that tracer binding is reversible. Compartment modelling revealed that all tracers were fitted best with the reversible two-tissue compartment model. Tracer uptake and distribution volume in lesions were in agreement with myelin status. However, the slow kinetics and homogeneous brain uptake of [{sup 11}C]CIC make this tracer less suitable for in vivo PET imaging. [{sup 11}C]PIB showed good uptake in the white matter in the cerebrum, but [{sup 11}C]PIB uptake in the cerebellum was low, despite high myelin density in this region. [{sup 11}C]MeDAS distribution correlated well with myelin density in different brain regions. This study showed that PET imaging of demyelination and remyelination processes in focal lesions is feasible. Our comparison of three myelin tracers showed that [{sup 11}C]MeDAS has more favourable properties for quantitative PET imaging of demyelinated and remyelinated lesions throughout the CNS than [{sup 11}C]CIC and [{sup 11}C]PIB. (orig.)

  5. 28 CFR 551.11 - Authority to approve a marriage.

    Science.gov (United States)

    2010-07-01

    ... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Authority to approve a marriage. 551.11... MISCELLANEOUS Marriages of Inmates § 551.11 Authority to approve a marriage. (a) The Warden may approve the marriage of a federal inmate confined in a federal institution. This authority may not be delegated below...

  6. Hydrolysis of lignocellulosic feedstock by novel cellulases originating from Pseudomonas sp. CL3 for fermentative hydrogen production.

    Science.gov (United States)

    Cheng, Chieh-Lun; Chang, Jo-Shu

    2011-09-01

    A newly isolated indigenous bacterium Pseudomonas sp. CL3 was able to produce novel cellulases consisting of endo-β-1,4-d-glucanase (80 and 100 kDa), exo-β-1,4-d-glucanase (55 kDa) and β-1,4-d-glucosidase (65 kDa) characterized by enzyme assay and zymography analysis. In addition, the CL3 strain also produced xylanase with a molecular weight of 20 kDa. The optimal temperature for enzyme activity was 50, 45, 45 and 55 °C for endo-β-1,4-d-glucanase, exo-β-1,4-d-glucanase, β-1,4-d-glucosidase and xylanase, respectively. All the enzymes displayed optimal activity at pH 6.0. The cellulases/xylanase could hydrolyze cellulosic materials very effectively and were thus used to hydrolyze natural agricultural waste (i.e., bagasse) for clean energy (H2) production by Clostridium pasteurianum CH4 using separate hydrolysis and fermentation process. The maximum hydrogen production rate and cumulative hydrogen production were 35 ml/L/h and 1420 ml/L, respectively, with a hydrogen yield of around 0.96 mol H2/mol glucose. Copyright © 2011 Elsevier Ltd. All rights reserved.

  7. Conversion of no-carrier-added [11C]carbon dioxide to [11C]carbon monoxide on molybdenum for the synthesis of 11C-labelled aromatic ketones

    International Nuclear Information System (INIS)

    Zeisler, S.K.; Nader, M.; Theobald, A.; Oberdorfer, F.

    1997-01-01

    A new method for the efficient conversion of no-carrier-added [ 11 C]carbon dioxide into [ 11 C]carbon monoxide is described. [ 11 C]Carbon dioxide produced by proton bombardment of ultra high purity nitrogen is pre-concentrated in a cryo trap and then passed through a quartz tube filled with a mesh of thin molybdenum wire heated to 850 o C. [ 11 C]Carbon dioxide readily reacts with molybdenum to form [ 11 C]carbon monoxide and molybdenum(IV) oxide. The latter also reduces carbon dioxide to carbon monoxide and helps improve the performance of the converter. [ 11 C]Carbon monoxide is purified from remaining [ 11 C]carbon dioxide and collected in a small silica trap from which it is eluted into a reaction mixture for the palladium-mediated synthesis of a 11 C-labelled aromatic ketone. Radiochemical yields of up to 81% (decay-corrected) for [ 11 C]carbon monoxide were obtained. Radiochemical purity and specific radioactivity of both [ 11 C]carbon monoxide and the 11 C-labelled ketone are sufficient for nuclear medical studies with PET. (Author)

  8. A radiometabolite study of the serotonin transporter PET radioligand [11C]MADAM

    International Nuclear Information System (INIS)

    Gourand, F.; Emond, P.; Bergström, J.P.; Takano, A.; Gulyás, B.; Guilloteau, D.; Barré, L.; Halldin, C.

    2014-01-01

    Introduction: 11 C]MADAM is a radioligand suitable for PET studies of the serotonin transporter (SERT). Metabolite analysis in human and non-human plasma samples using HPLC separation has shown that [ 11 C]MADAM was rapidly metabolized. A possible metabolic pathway is the S-oxidation which could lead to SOMADAM and SO 2 MADAM. In vitro evaluation of these two potential metabolites has shown that SOMADAM exhibited a good affinity for SERT and a good selectivity for SERT over NET and DAT. Methods: Comparative PET imaging studies in non-human primate brain with [ 11 C]MADAM and [ 11 C]SOMADAM were carried out, and plasma samples were analyzed using reverse phase HPLC. We have explored the metabolism of [ 11 C]MADAM in rat brain with a view to understand its possible interference for brain imaging with PET. Results: PET imaging studies in non-human primate brain using [ 11 C]SOMADAM indicated that this tracer does not bind with high amounts to brain regions known to be rich in SERT. The fraction of [ 11 C]SOMADAM in non-human primate plasma was approximately 5% at 4 min and 1% at 15 min after [ 11 C]MADAM injection. HPLC analysis of brain sample after [ 11 C]MADAM injection to rats demonstrated that [ 11 C]SOMADAM was not detected in the brain. Conclusions: 11 C]SOMADAM is not superior over [ 11 C]MADAM as a SERT PET radioligand. Nevertheless, [ 11 C]SOMADAM has been identified as a minor labeled metabolite of [ 11 C]MADAM measured in monkey plasma. [ 11 C]SOMADAM was not detected in rat brain

  9. 11 CFR 100.11 - State (2 U.S.C. 431(12)).

    Science.gov (United States)

    2010-01-01

    ... 11 Federal Elections 1 2010-01-01 2010-01-01 false State (2 U.S.C. 431(12)). 100.11 Section 100.11 Federal Elections FEDERAL ELECTION COMMISSION GENERAL SCOPE AND DEFINITIONS (2 U.S.C. 431) General Definitions § 100.11 State (2 U.S.C. 431(12)). State means each State of the United States, the District of...

  10. Revalidation of the NASA Ames 11-by 11-Foot Transonic Wind Tunnel with a Commercial Airplane Model

    Science.gov (United States)

    Kmak, Frank J.; Hudgins, M.; Hergert, D.; George, Michael W. (Technical Monitor)

    2001-01-01

    The 11-By 11-Foot Transonic leg of the Unitary Plan Wind Tunnel (UPWT) was modernized to improve tunnel performance, capability, productivity, and reliability. Wind tunnel tests to demonstrate the readiness of the tunnel for a return to production operations included an Integrated Systems Test (IST), calibration tests, and airplane validation tests. One of the two validation tests was a 0.037-scale Boeing 777 model that was previously tested in the 11-By 11-Foot tunnel in 1991. The objective of the validation tests was to compare pre-modernization and post-modernization results from the same airplane model in order to substantiate the operational readiness of the facility. Evaluation of within-test, test-to-test, and tunnel-to-tunnel data repeatability were made to study the effects of the tunnel modifications. Tunnel productivity was also evaluated to determine the readiness of the facility for production operations. The operation of the facility, including model installation, tunnel operations, and the performance of tunnel systems, was observed and facility deficiency findings generated. The data repeatability studies and tunnel-to-tunnel comparisons demonstrated outstanding data repeatability and a high overall level of data quality. Despite some operational and facility problems, the validation test was successful in demonstrating the readiness of the facility to perform production airplane wind tunnel%, tests.

  11. Effect of amount of straw provided to growing/finishing pigs on gastric ulceration at slaughter

    DEFF Research Database (Denmark)

    Herskin, Mette S.; Jensen, Henrik Elvang; Jespersen, Anna

    2014-01-01

    The effect of a current commercial xylanase (DAN) and experimental xylanase (EX), and EX in combination with protease (EXP), on the degradation and apparent ileal digestibility (AID) of non-starch polysaccharides (NSP) in wheat Distillers Dried Grains with Solubles (DDGS), was studied in 8 ileum......-cannulated pigs (initial BW 36.6±2.8 kg) following a double 4x4 Latin Square design. The control and three enzyme diets, each containing 96% DDGS, were supplemented with vitamins, minerals, L-lysine, 500 FTU phytase/kg feed, dust-binder and chromic oxide (3 g/kg). The pigs were fed 3 times daily for 1 week...

  12. The structure of the TFIIH p34 subunit reveals a von Willebrand factor A like fold.

    Directory of Open Access Journals (Sweden)

    Dominik R Schmitt

    Full Text Available RNA polymerase II dependent transcription and nucleotide excision repair are mediated by a multifaceted interplay of subunits within the general transcription factor II H (TFIIH. A better understanding of the molecular structure of TFIIH is the key to unravel the mechanism of action of this versatile protein complex within these vital cellular processes. The importance of this complex becomes further evident in the context of severe diseases like xeroderma pigmentosum, Cockayne's syndrome and trichothiodystrophy, that arise from single point mutations in TFIIH subunits. Here we describe the structure of the p34 subunit of the TFIIH complex from the eukaryotic thermophilic fungus Chaetomium thermophilum. The structure revealed that p34 contains a von Willebrand Factor A (vWA like domain, a fold which is generally known to be involved in protein-protein interactions. Within TFIIH p34 strongly interacts with p44, a positive regulator of the helicase XPD. Putative protein-protein interfaces are analyzed and possible binding sites for the p34-p44 interaction suggested.

  13. Educational Modelling Language 1.1 (EML 1.1)

    NARCIS (Netherlands)

    Koper, Rob; Vogten, Hubert; Martens, Harrie

    2003-01-01

    Package contains the EML 1.1 dtd, schema and documentation as is created for the Edubox Player. It is slightly different from IMS Learning Design and provides an intermediate solution between EML 1.0 and IMD Learning Design. The Edubox Player 3.x works on EML 1.1. This package is stored for

  14. Experience in the design, implementation and use of PL-11, a programming language for the PDP-11

    CERN Document Server

    Russell, R D

    1976-01-01

    PL-11 is a programming language for the PDP-11 family of computers designed and implemented as part of the OMEGA Project at CERN (the European Organization for Nuclear Research). Its purpose is to provide an effective tool for both physicists and systems programmers to use in building real time data acquisition systems that are online to high-energy physics experiments. It is a fairly typical member of the PL-class of programming languages which are based on the initial design of PL360. (44 refs).

  15. 34 CFR 686.11 - Eligibility to receive a grant.

    Science.gov (United States)

    2010-07-01

    ... 34 Education 3 2010-07-01 2010-07-01 false Eligibility to receive a grant. 686.11 Section 686.11 Education Regulations of the Offices of the Department of Education (Continued) OFFICE OF POSTSECONDARY EDUCATION, DEPARTMENT OF EDUCATION TEACHER EDUCATION ASSISTANCE FOR COLLEGE AND HIGHER EDUCATION (TEACH...

  16. Hidrología, 2010-11

    OpenAIRE

    Sánchez San Román, Francisco Javier

    2010-01-01

    I. Materiales de clase: 1. El Ciclo Hidrológico. 2. Precipitaciones. 3. Evapotranspiración. 4. Hidrología Superficial (I): Medidas y Tratamiento de los datos. 5. Hidrología Superficial (II): Hidrogramas. 6. Hidrología Superficial (III): Relación Precipitación - Escorrentía. 7. Conceptos Fundamentales de Hidrogeología. 8. Flujo en medios porosos: Ley de Darcy. 9. Hidráulica Subterránea: Principios Básicos. 10. Hidráulica de captaciones: Fundamentos. 11. Hidroquímica. Conceptos Fundamentales. 1...

  17. Hoarding disorder: a new diagnostic category in ICD-11?

    Directory of Open Access Journals (Sweden)

    Leonardo F. Fontenelle

    2014-01-01

    Full Text Available Despite the long-held view that hoarding is a symptom of both obsessive-compulsive disorder and obsessive-compulsive personality disorder, increased evidence has emerged during the last 20 years suggesting that hoarding represents a distinct form of psychopathology. This study reflects the discussions on the nosological status of hoarding carried out by the WHO ICD-11 Working Group on the Classification of Obsessive-Compulsive and Related Disorders. The distinctiveness of hoarding is based on its having core symptoms that differ from those of other disorders, as well as distinctive neurobiological correlates and treatment responses. Furthermore, data showing the clinical utility, global applicability, and appropriateness of the concept of hoarding disorder outside specialty mental health settings suggest that this condition should be included in ICD-11. Finally, given the focus of ICD-11 on primary care and public health, the Working Group suggests that poor insight and severe domestic squalor may be considered as specifiers for hoarding disorder in ICD-11.

  18. 25 CFR 11.1007 - Petition.

    Science.gov (United States)

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false Petition. 11.1007 Section 11.1007 Indians BUREAU OF... Juvenile Offender Procedure § 11.1007 Petition. (a) Proceedings under §§ 11.1000-11.1014 of this part shall be instituted by a petition filed by the presenting officer on behalf of the tribe and in the...

  19. Total synthesis of [2-11C]thymidine from [11C]urea: A tracer of choice for measurement of cellular proliferation using PET

    International Nuclear Information System (INIS)

    Labar, D.; Vander Borght, T.

    1990-01-01

    In preliminary studies of cellular proliferation with [methyl- 11 C]thymidine, the labelled degradative products mask the progressive incorporation of the tracer into DNA. The authors have developed a procedure for the synthesis of [2- 11 C]thymidine to circumvent this difficulty, using a [ 11 C]urea precursor

  20. Reaction of 11 C-benzoyl chlorides with metalloid reagents: 11 C-labeling of benzyl alcohols, benzaldehydes, and phenyl ketones from [11 C]CO.

    Science.gov (United States)

    Roslin, Sara; Dahl, Kenneth; Nordeman, Patrik

    2018-01-26

    In this article, we describe the carbon-11 ( 11 C, t 1/2  = 20.4 minutes) labeling of benzyl alcohols, benzaldehydes, and ketones using an efficient 2-step synthesis in which 11 C-carbon monoxide is used in an initial palladium-mediated reaction to produce 11 C-benzoyl chloride as a key intermediate. In the second step, the obtained 11 C-benzoyl chloride is further treated with a metalloid reagent to furnish the final 11 C-labeled product. Benzyl alcohols were obtained in moderated to high non-isolated radiochemical yields (RCY, 35%-90%) with lithium aluminum hydride or lithium aluminum deuteride as metalloid reagent. Changing the metalloid reagent to either tributyltin hydride or sodium borohydride, allowed for the reliable syntheses of 11 C-benzaldehydes in RCYs ranging from 58% to 95%. Finally, sodium tetraphenylborate were utilized to obtain 11 C-phenyl ketones in high RCYs (77%-95%). The developed method provides a new and efficient route to 3 different classes of compounds starting from aryl iodides or aryl bromides. Copyright © 2018 John Wiley & Sons, Ltd.

  1. Drosophila SLC5A11 Mediates Hunger by Regulating K(+) Channel Activity.

    Science.gov (United States)

    Park, Jin-Yong; Dus, Monica; Kim, Seonil; Abu, Farhan; Kanai, Makoto I; Rudy, Bernardo; Suh, Greg S B

    2016-08-08

    Hunger is a powerful drive that stimulates food intake. Yet, the mechanism that determines how the energy deficits that result in hunger are represented in the brain and promote feeding is not well understood. We previously described SLC5A11-a sodium/solute co-transporter-like-(or cupcake) in Drosophila melanogaster, which is required for the fly to select a nutritive sugar over a sweeter nonnutritive sugar after periods of food deprivation. SLC5A11 acts on approximately 12 pairs of ellipsoid body (EB) R4 neurons to trigger the selection of nutritive sugars, but the underlying mechanism is not understood. Here, we report that the excitability of SLC5A11-expressing EB R4 neurons increases dramatically during starvation and that this increase is abolished in the SLC5A11 mutation. Artificial activation of SLC5A11-expresssing neurons is sufficient to promote feeding and hunger-driven behaviors; silencing these neurons has the opposite effect. Notably, SLC5A11 transcript levels in the brain increase significantly when flies are starved and decrease shortly after starved flies are refed. Furthermore, expression of SLC5A11 is sufficient for promoting hunger-driven behaviors and enhancing the excitability of SLC5A11-expressing neurons. SLC5A11 inhibits the function of the Drosophila KCNQ potassium channel in a heterologous expression system. Accordingly, a knockdown of dKCNQ expression in SLC5A11-expressing neurons produces hunger-driven behaviors even in fed flies, mimicking the overexpression of SLC5A11. We propose that starvation increases SLC5A11 expression, which enhances the excitability of SLC5A11-expressing neurons by suppressing dKCNQ channels, thereby conferring the hunger state. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Synthesis of [11C]cyanoalkyltriphenylphosphoranes via [11C]cyanide substitution on haloalkylphosphonium salts

    International Nuclear Information System (INIS)

    Hoernfeldt, K.

    1994-01-01

    The synthesis of the 11 C-labelled bifunctional precursors 3-[ 11 C]cyanoethyltriphenylphosphonium bromide (1'), 4-[ 11 C]cyanopropyltriphenylphosphonium bromide (2'), 5-[ 11 C]cyanobutyltriphenylphosphonium bromide (3'), 4-[ 11 C]cyanopropyltriphenylphosphonium iodide (4') and 5-[ 11 C]cyanobutyltriphenylphosphonium iodide (5') is presented. The label was introduced using [ 11 C]cyanide in a substitution reaction on the ω-halo-alkyltriphenylphosphonium salt (bromide or iodide salt). The phosphonium salts 1'-5' were formed in 33-99% radiochemical yield in 5-10 min reaction time. After addition of epichlorohydrin as generator of base, the precursors 1-3 were formed. The potential of the intermediates 1-3 in Wittig reactions was shown in model reactions with aromatic and aliphatic aldehydes. The aromatic olefins obtained from 1'-5' were formed in 85-96% radiochemical yield, with Z/E ratios between 67/33-75/25. The aliphatic olefins were obtained in 60-78% radiochemical yield from 4' and 5'. In the reaction with 1' and an aliphatic aldehyde, the yield decreased to 5-10%. The Z/E ratios were 100/0 for the aliphatic olefins. In an experiment starting with 2,7 GBq (73 mCi) hydrogen [ 11 C]cyanide, 451 MBq (12.2 mCi) olefin from 3' and 4-nitrobenzaldehyde was obtained in 44 min from hydrogen [ 11 C]cyanide production, with a 55% decay corrected radiochemical yield, the radiochemical purity was 96%. (Author)

  3. C-11 cyanide production system

    Science.gov (United States)

    Kim, Dohyun; Alexoff, David; Kim, Sung Won; Hooker, Jacob M.; Ferrieri, Richard A.

    2017-11-21

    A method for providing .sup.11C-labeled cyanides from .sup.11C labeled oxides in a target gas stream retrieved from an irradiated high pressure gaseous target containing O.sub.2, wherein .sup.11C labeled oxides are reduced with H.sub.2 in the presence of a nickel catalyst under a pressure and a temperature sufficient to form a product stream comprising at least about 95% .sup.11CH.sub.4, the .sup.11CH.sub.4 is then combined with an excess of NH.sub.3 in a carrier/reaction stream flowing at an accelerated velocity and the combined .sup.11CH4 carrier/reaction stream is then contacted with a platinum (Pt) catalyst particulate supported on a substantially-chemically-nonreactive heat-stable support at a temperature of at least about 900.degree. C., whereby a product stream comprising at least about 60% H.sup.11CN is provided in less than 10 minutes from retrieval of the .sup.11C labeled oxide.

  4. A self-report measure for the ICD-11 dimensional trait model proposal: The personality inventory for ICD-11.

    Science.gov (United States)

    Oltmanns, Joshua R; Widiger, Thomas A

    2018-02-01

    Proposed for the 11th edition of the World Health Organization's International Classification of Diseases (ICD-11) is a dimensional trait model for the classification of personality disorder (Tyrer, Reed, & Crawford, 2015). The ICD-11 proposal consists of 5 broad domains: negative affective, detachment, dissocial, disinhibition, and anankastic (Mulder, Horwood, Tyrer, Carter, & Joyce, 2016). Several field trials have examined this proposal, yet none has included a direct measure of the trait model. The purpose of the current study was to develop and provide initial validation for the Personality Inventory for ICD-11 (PiCD), a self-report measure of this proposed 5-domain maladaptive trait model. Item selection and scale construction proceeded through 3 initial data collections assessing potential item performance. Two subsequent studies were conducted for scale validation. In Study 1, the PiCD was evaluated in a sample of 259 MTurk participants (who were or had been receiving mental health treatment) with respect to 2 measures of general personality structure: The Eysenck Personality Questionnaire-Revised and the 5-Dimensional Personality Test. In Study 2, the PiCD was evaluated in an additional sample of 285 participants with respect to 2 measures of maladaptive personality traits: The Personality Inventory for DSM-5 and the Computerized Adaptive Test for Personality Disorders. Study 3 provides an item-level exploratory structural equation model with the combined samples from Studies 1 and 2. The results are discussed with respect to the validity of the measure and the potential benefits for future research in having a direct, self-report measure of the ICD-11 trait proposal. (PsycINFO Database Record (c) 2018 APA, all rights reserved).

  5. Duplication of 17(p11.2p11.2) in a male child with autism and severe language delay.

    Science.gov (United States)

    Nakamine, Alisa; Ouchanov, Leonid; Jiménez, Patricia; Manghi, Elina R; Esquivel, Marcela; Monge, Silvia; Fallas, Marietha; Burton, Barbara K; Szomju, Barbara; Elsea, Sarah H; Marshall, Christian R; Scherer, Stephen W; McInnes, L Alison

    2008-03-01

    Duplications of 17(p11.2p11.2) have been associated with various behavioral manifestations including attention deficits, obsessive-compulsive symptoms, autistic traits, and language delay. We are conducting a genetic study of autism and are screening all cases for submicroscopic chromosomal abnormalities, in addition to standard karyotyping, and fragile X testing. Using array-based comparative genomic hybridization analysis of data from the Affymetrix GeneChip(R) Human Mapping Array set, we detected a duplication of approximately 3.3 Mb on chromosome 17p11.2 in a male child with autism and severe expressive language delay. The duplication was confirmed by measuring the copy number of genomic DNA using quantitative polymerase chain reaction. Gene expression analyses revealed increased expression of three candidate genes for the Smith-Magenis neurobehavioral phenotype, RAI1, DRG2, and RASD1, in transformed lymphocytes from Case 81A, suggesting gene dosage effects. Our results add to a growing body of evidence suggesting that duplications of 17(p11.2p11.2) result in language delay as well as autism and related phenotypes. As Smith-Magenis syndrome is also associated with language delay, a gene involved in acquisition of language may lie within this interval. Whether a parent of origin effect, gender of the case, the presence of allelic variation, or changes in expression of genes outside the breakpoints influence the resultant phenotype remains to be determined. (c) 2007 Wiley-Liss, Inc.

  6. Enzymes improve ECF bleaching of pulp

    Directory of Open Access Journals (Sweden)

    Lachenal, D.

    2006-07-01

    Full Text Available The delignification efficiency of different laccase enzymes was examined on the eucalyptus Kraft pulp. The laccase enzyme from Trametes versicolor showing the highest delignification efficiency was selected and used in the elemental chlorine-free bleaching sequence for improving the pulp bleachability. An appreciable reduction in chlorine dioxide consumption was also obtained. Further reduction in chlorine dioxide consumption was obtained when the same laccase treated pulp was subjected to an acid treatment after the extraction stage followed by the DEPD sequence. Elemental-chlorine free bleaching was also performed using the xylanase-laccase treated pulp. Xylanase treatment was incorporated to the laccase mediator system in the elemental-chlorine free bleaching both sequentially and simultaneously. The bleaching sequence DEPD followed and in both the cases, the reduction in chlorine dioxide consumption was greater in comparison to the control. The chlorine dioxide consumption was reduced further when xylanase-laccase treated pulp was given an additional acid treatment. The final pulp properties of the treated pulps were comparable to the control pulp.

  7. Automatic synthesis of [11C]NKY-722 with high specific activity, using anhydrous [11C] methanol as a precursor

    International Nuclear Information System (INIS)

    Suzuki, Kazutoshi; Inoue, Osamu; Itoh, Takashi; Nemoto, Kazuyoshi; Oosumi, Seimei; Miwa, Soichi.

    1992-01-01

    3-(4-allyl-1-piperazinyl)-2,2-dimethylpropyl methyl 1,4-dihydro-2,6-dimethyl- 4-(3-nitrophenyl)-3,5-pyridine dicarboxylate (NKY-722) was labeled with carbon-11 using anhydrous [ 11 C] methanol. Using a computer controlled equipment, a few GBq of [ 11 C] NKY-722 with the specific activity of 120 - 180 GBq/μmol could by synthesized at the radiochemical purity of > 99% in 10 ml of physiological saline containing Polysolvate-80 (1.5 vol%) and ethyl alcohol (0.75 vol%). Preliminary PET experiments using rats and a rhesus monkey have bee done, and very low accumulation of the compound into the brain, however comparatively higher accumulation in the heart were observed. (author)

  8. The IEEE 802.11a standards

    Indian Academy of Sciences (India)

    First page Back Continue Last page Overview Graphics. The IEEE 802.11a standards. Provides higher data rate and system capacities and uses OFDM in Physical Layer to mitigate the multi path effects;. Supports multiple 20Mhz channel. Each channel being an OFDM Modulated;; 52 Carriers. 48 data carrier; 4 Carry pilot ...

  9. 42 CFR 1008.11 - Who may submit a request.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 5 2010-10-01 2010-10-01 false Who may submit a request. 1008.11 Section 1008.11 Public Health OFFICE OF INSPECTOR GENERAL-HEALTH CARE, DEPARTMENT OF HEALTH AND HUMAN SERVICES OIG... advisory opinion regarding an existing arrangement or one which the requestor in good faith specifically...

  10. Reliable set-up for in-loop 11C-carboxylations using Grignard reactions for the preparation of [carbonyl-11C]WAY-100635 and [11C]-(+)-PHNO

    International Nuclear Information System (INIS)

    Rami-Mark, Christina; Ungersboeck, Johanna; Haeusler, Daniela; Nics, Lukas; Philippe, Cecile; Mitterhauser, Markus; Willeit, Matthaeus; Lanzenberger, Rupert; Karanikas, Georgios; Wadsak, Wolfgang

    2013-01-01

    Aim of this work was the implementation of a generalized in-loop synthesis for 11 C-carboxylations and subsequent 11 C-acylations on the TRACERlab FxC Pro platform. The set-up was tested using [carbonyl- 11 C]WAY-100635 and, for the first time, [ 11 C]-(+)-PHNO. Its general applicability could be demonstrated and both [carbonyl- 11 C]WAY-100635 and [ 11 C]-(+)-PHNO were prepared with high reliability and satisfying outcome. - Highlights: • Generalized method for in-loop 11 C-carboxylations implemented. • Grignard reactions successfully tested. • First in-loop procedure for [ 11 C]-(+)PHNO established. • Satisfactory synthesis outcome for both [carbonyl- 11 C]WAY-100635 and [ 11 C]-(+)PHNO. • No distillation for purification of intermediate required

  11. A Novel Mutation in the CYP11B1 Gene Causes Steroid 11β-Hydroxylase Deficient Congenital Adrenal Hyperplasia with Reversible Cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Mohammad A. Alqahtani

    2015-01-01

    Full Text Available Congenital adrenal hyperplasia (CAH due to steroid 11β-hydroxylase deficiency is the second most common form of CAH, resulting from a mutation in the CYP11B1 gene. Steroid 11β-hydroxylase deficiency results in excessive mineralcorticoids and androgen production leading to hypertension, precocious puberty with acne, enlarged penis, and hyperpigmentation of scrotum of genetically male infants. In the present study, we reported 3 male cases from a Saudi family who presented with penile enlargement, progressive darkness of skin, hypertension, and cardiomyopathy. The elder patient died due to heart failure and his younger brothers were treated with hydrocortisone and antihypertensive medications. Six months following treatment, cardiomyopathy disappeared with normal blood pressure and improvement in the skin pigmentation. The underlying molecular defect was investigated by PCR-sequencing analysis of all coding exons and intron-exon boundary of the CYP11B1 gene. A novel biallelic mutation c.780 G>A in exon 4 of the CYP11B1 gene was found in the patients. The mutation created a premature stop codon at amino acid 260 (p.W260∗, resulting in a truncated protein devoid of 11β-hydroxylase activity. Interestingly, a somatic mutation at the same codon (c.779 G>A, p.W260∗ was reported in a patient with papillary thyroid cancer (COSMIC database. In conclusion, we have identified a novel nonsense mutation in the CYP11B1 gene that causes classic steroid 11β-hydroxylase deficient CAH. Cardiomyopathy and cardiac failure can be reversed by early diagnosis and treatment.

  12. Partial deletion 11q

    DEFF Research Database (Denmark)

    Hertz, Jens Michael; Tommerup, N; Sørensen, F B

    1995-01-01

    We describe the cytogenetic findings and the dysmorphic features in a stillborn girl with a large de novo terminal deletion of the long arm of chromosome 11. The karyotype was 46,XX,del(11)(q21qter). By reviewing previous reports of deletion 11q, we found that cleft lip and palate are most...

  13. Monitorando a degradação da poliamida 11 (PA-11 via espectroscopia na região do infravermelho médio com transformada de fourier (FTIR Monitoring the degradation of polyamide 11 (PA-11 via fourier transform infrared spectroscopy (FTIR

    Directory of Open Access Journals (Sweden)

    Eloilson Domingos

    2012-01-01

    Full Text Available O potencial da técnica de espectroscopia de infravermelho com transformada de Fourier e acessório de reflexão total atenuada (FTIR-ATR foi avaliado para o monitoramento da degradação da poliamida 11 (PA-11 usada em dutos flexíveis. As amostras foram submetidas a envelhecimento em reatores com pressão controlada e atmosfera inerte. Os corpos de prova foram imersos em água deionizada (pH 7, nas temperaturas de 110, 120 e 140 ºC por um período de até 50 dias. A técnica recomendada para monitoramento da degradação da PA-11 é a viscosimetria, através de medidas da viscosidade inerente corrigida (VIC e a análise termogravimétrica (TGA. O comportamento observado para a VIC e TGA durante o envelhecimento possibilitou a construção de modelos que correlacionam com a técnica de FTIR-ATR. A partir dos resultados de FTIR-ATR, a variação na intensidade da banda atribuída à fase amorfa, 1161 cm-1, possibilitou o monitoramento do envelhecimento quando associamos a técnica a modelos quimiométricos como o de regressão dos mínimos quadrados parciais, PLS. Portanto, a técnica FTIR-ATR pode ser uma nova alternativa no monitoramento da degradação hidrolítica da PA-11, eliminando assim o uso de solventes orgânicos tóxicos e reduzindo, consequentemente, o tempo de análise.The potential of the infrared spectroscopy with the attenuated total reflection technique (FTIR-ATR was evaluated to monitor degradation of polyamide 11, PA-11, applied in flexible pipes. The samples were subjected to aging on reactors with controlled pressure and atmosphere. The samples were immersed in deionized water, pH 7, at temperatures of 110, 120 and 140 ºC over a period of up to 50 days. The typical technique recommended for monitoring PA-11 degradation is viscometry from inherent viscosity corrected (VIC and thermogravimetric (TGA measurements. TGA and VIC results allowed the use of chemometric models that can be related to FTIR-ATR spectra, with

  14. Monitorando a degradação da poliamida 11 (PA-11 via espectroscopia na região do infravermelho médio com transformada de fourier (FTIR Monitoring the degradation of polyamide 11 (PA-11 via fourier transform infrared spectroscopy (FTIR

    Directory of Open Access Journals (Sweden)

    Eloilson Domingos

    2013-01-01

    Full Text Available O potencial da técnica de espectroscopia de infravermelho com transformada de Fourier e acessório de reflexão total atenuada (FTIR-ATR foi avaliado para o monitoramento da degradação da poliamida 11 (PA-11 usada em dutos flexíveis. As amostras foram submetidas a envelhecimento em reatores com pressão controlada e atmosfera inerte. Os corpos de prova foram imersos em água deionizada (pH 7, nas temperaturas de 110, 120 e 140 ºC por um período de até 50 dias. A técnica recomendada para monitoramento da degradação da PA-11 é a viscosimetria, através de medidas da viscosidade inerente corrigida (VIC e a análise termogravimétrica (TGA. O comportamento observado para a VIC e TGA durante o envelhecimento possibilitou a construção de modelos que correlacionam com a técnica de FTIR-ATR. A partir dos resultados de FTIR-ATR, a variação na intensidade da banda atribuída à fase amorfa, 1161 cm-1, possibilitou o monitoramento do envelhecimento quando associamos a técnica a modelos quimiométricos como o de regressão dos mínimos quadrados parciais, PLS. Portanto, a técnica FTIR-ATR pode ser uma nova alternativa no monitoramento da degradação hidrolítica da PA-11, eliminando assim o uso de solventes orgânicos tóxicos e reduzindo, consequentemente, o tempo de análise.The potential of the infrared spectroscopy with the attenuated total reflection technique (FTIR-ATR was evaluated to monitor degradation of polyamide 11, PA-11, applied in flexible pipes. The samples were subjected to aging on reactors with controlled pressure and atmosphere. The samples were immersed in deionized water, pH 7, at temperatures of 110, 120 and 140 ºC over a period of up to 50 days. The typical technique recommended for monitoring PA-11 degradation is viscometry from inherent viscosity corrected (VIC and thermogravimetric (TGA measurements. TGA and VIC results allowed the use of chemometric models that can be related to FTIR-ATR spectra, with

  15. Construction and characterization of the interdomain chimeras using Cry11Aa and Cry11Ba from Bacillus thuringiensis and identification of a possible novel toxic chimera.

    Science.gov (United States)

    Sun, Yunjun; Zhao, Qiang; Zheng, Dasheng; Ding, Xuezhi; Wang, Jingfang; Hu, Quanfang; Yuan, Zhiming; Park, Hyun-Woo; Xia, Liqiu

    2014-01-01

    Three structural domains of mosquitocidal Cry11Aa and Cry11Ba from Bacillus thuringiensis were exchanged to produce interdomain chimeras [BAA (11Ba/11Aa/11Aa), ABA (11Aa/11Ba/11Aa), AAB (11Aa/11Aa/11Ba), ABB (11Aa/11Ba/11Ba), BAB (11Ba/11Aa/11Ba), BBA (11Ba/11Ba/11Aa]. Chimeras BAB, BAA, BBA, and AAB formed inclusion bodies in the crystal-negative B. thuringiensis host and produced expected protein bands on SDS-PAGE gel. However, no inclusion body or target protein could be found for chimeras ABA and ABB. In bioassays using the fourth-instar larvae of Culex quinquefasciatus and Aedes aegypti, AAB had ~50 % lethal concentrations of 4.8 and 2.2 μg ml(-1), respectively; however, the rest of chimeras were not toxic. This study thus helps to understand the domain-function relationships of the Cry11Aa and Cry11Ba toxins. The toxic chimera, AAB, might be a candidate for mosquito control as its amino acid sequence is different from the two parental toxins.

  16. Use of a new Trichoderma harzianum strain isolated from the Amazon rainforest with pretreated sugar cane bagasse for on-site cellulase production.

    Science.gov (United States)

    Delabona, Priscila da Silva; Farinas, Cristiane Sanchez; da Silva, Mateus Ribeiro; Azzoni, Sindelia Freitas; Pradella, José Geraldo da Cruz

    2012-03-01

    The on-site production of cellulases is an important strategy for the development of sustainable second-generation ethanol production processes. This study concerns the use of a specific cellulolytic enzyme complex for hydrolysis of pretreated sugar cane bagasse. Glycosyl hydrolases (FPase, xylanase, and β-glucosidase) were produced using a new strain of Trichoderma harzianum, isolated from the Amazon rainforest and cultivated under different conditions. The influence of the carbon source was first investigated using shake-flask cultures. Selected carbon sources were then further studied under different pH conditions using a stirred tank bioreactor. Enzymatic activities up to 121 FPU/g, 8000 IU/g, and 1730 IU/g of delignified steam-exploded bagasse+sucrose were achieved for cellulase, xylanase and β-glucosidase, respectively. This enzymatic complex was used to hydrolyze pretreated sugar cane bagasse. A comparative evaluation, using an enzymatic extract from Trichoderma reesei RUTC30, indicated similar performance of the T. harzianum enzyme complex, being a potential candidate for on-site production of enzymes. Copyright © 2011 Elsevier Ltd. All rights reserved.

  17. 25 CFR 11.1107 - Petition.

    Science.gov (United States)

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false Petition. 11.1107 Section 11.1107 Indians BUREAU OF... Minor-in-Need-of-Care Procedure § 11.1107 Petition. Proceedings under §§ 11.1100-11.1114 of this part shall be instituted by a petition filed by the presenting officer on behalf of the tribe and the...

  18. Tracers development for the PET study of nicotinic receptors: [{sup 11}C]-mecamylamine and [{sup 11}C]-SIB 1553A. Tritium and carbon-11 radiolabelling of a serine proteinase inhibitor: the t-PA{sub stop}; Developpement de traceurs pour l'etude des recepteurs nicotiniques par TEP: la [{sup 11}C]-mecamylamine et le [{sup 11}C]SIB 1553A. Radiomarquages par le tritium et le carbone-11 d'un inhibiteur d'une serine protease: le t-PA{sub stop}

    Energy Technology Data Exchange (ETDEWEB)

    Sobrio, F.

    2002-12-15

    In order to develop radiotracers for the Positron Emission Tomography (PET), we labelled both the mecamylamine and SIB-1553A with carbon-11 to study the nicotinic cholinergic receptors (nAChRs). The radiosynthesis of [{sup 11}C]-t-PA{sub stop} and the labelling with tritium of one analogue were realized for cerebral ischemia PET studies. The [{sup 11}C]-mecamylamine, a non-competitive and non-selective nAChRs antagonist was synthesized in 45 min via a N-[{sup 11}C]-methylation reaction. In the rat brain, the ex vivo studies showed no radio-metabolite 45 min after the injection of [{sup 11}C]-mecamylamine. The uptake kinetics in the rat brain or in vivo by PET in the anesthetized baboon or in the conscious monkey, reached a plateau around 45-50 min after injection. However, the saturation or displacement experiments did not permit to exhibit nor a significant difference of labelling between the different cerebral regions nor a specific uptake. In consequence, the [{sup 11}C]-mecamylamine was not an appropriate radioligand for nAChRs PET study. The labelling of [{sup 11}C]-SIB 1553A, a selective agonist for the nicotinic {beta}4 subunit, required the synthesis in 5 steps (56% overall yield) of precursor for the incorporation of carbon-11. The radiosynthesis was performed in 36 min by a N-[{sup 11}C]-methylation reaction (yield: 75%). The [{sup 11}C]-t-PA{sub stop} was obtained from [{sup 11}C]-KCN with yields from 80 to 90%. For the first time with carbon-11, the formation of an amidine group was realized from a nitrile group. The labelling by isotopic exchange of hydrogen by tritium of the t-PA{sub stop} did not permit to obtain the [{sup 3}H]-t-PA{sub stop} but a tritiated analogue. This compound will be used to study its vectorization by micro-encapsulation. (author)

  19. Purification, crystallization and preliminary X-ray analysis of a thermostable glycoside hydrolase family 43 β-xylosidase from Geobacillus thermoleovorans IT-08

    International Nuclear Information System (INIS)

    Rohman, Ali; Oosterwijk, Niels van; Kralj, Slavko; Dijkhuizen, Lubbert; Dijkstra, Bauke W.; Puspaningsih, Ni Nyoman Tri

    2007-01-01

    The β-xylosidase was crystallized using PEG 6000 as precipitant. 5% PEG 6000 yielded bipyramid-shaped tetragonal crystals diffracting to 1.55 Å resolution, and 13% PEG 6000 gave rectangular monoclinic crystals diffracting to 1.80 Å resolution. The main enzymes involved in xylan-backbone hydrolysis are endo-1,4-β-xylanase and β-xylosidase. β-Xylosidase converts the xylo-oligosaccharides produced by endo-1,4-β-xylanase into xylose monomers. The β-xylosidase from the thermophilic Geobacillus thermoleovorans IT-08, a member of glycoside hydrolase family 43, was crystallized at room temperature using the hanging-drop vapour-diffusion method. Two crystal forms were observed. Bipyramid-shaped crystals belonging to space group P4 3 2 1 2, with unit-cell parameters a = b = 62.53, c = 277.4 Å diffracted to 1.55 Å resolution. The rectangular crystals belonged to space group P2 1 , with unit-cell parameters a = 57.94, b = 142.1, c = 153.9 Å, β = 90.5°, and diffracted to 1.80 Å resolution

  20. 31 CFR 375.11 - Who may participate in a redemption operation?

    Science.gov (United States)

    2010-07-01

    ... Bank of New York has approved to conduct open market transactions with the Bank. (b) Others. A person... operation? 375.11 Section 375.11 Money and Finance: Treasury Regulations Relating to Money and Finance... SECURITIES REDEMPTION OPERATIONS Offering, Certifications, and Delivery § 375.11 Who may participate in a...

  1. Aquaporin-11: A channel protein lacking apparent transport function expressed in brain

    Directory of Open Access Journals (Sweden)

    Tsunenari Takashi

    2006-05-01

    Full Text Available Abstract Background The aquaporins are a family of integral membrane proteins composed of two subfamilies: the orthodox aquaporins, which transport only water, and the aquaglyceroporins, which transport glycerol, urea, or other small solutes. Two recently described aquaporins, numbers 11 and 12, appear to be more distantly related to the other mammalian aquaporins and aquaglyceroporins. Results We report on the characterization of Aquaporin-11 (AQP11. AQP11 RNA and protein is found in multiple rat tissues, including kidney, liver, testes and brain. AQP11 has a unique distribution in brain, appearing in Purkinje cell dendrites, hippocampal neurons of CA1 and CA2, and cerebral cortical neurons. Immunofluorescent staining of Purkinje cells indicates that AQP11 is intracellular. Unlike other aquaporins, Xenopus oocytes expressing AQP11 in the plasma membrane failed to transport water, glycerol, urea, or ions. Conclusion AQP11 is functionally distinct from other proteins of the aquaporin superfamily and could represent a new aquaporin subfamily. Further studies are necessary to elucidate the role of AQP11 in the brain.

  2. 37 CFR 11.26 - Settlement.

    Science.gov (United States)

    2010-07-01

    ... 37 Patents, Trademarks, and Copyrights 1 2010-07-01 2010-07-01 false Settlement. 11.26 Section 11... Disciplinary Proceedings; Jurisdiction, Sanctions, Investigations, and Proceedings § 11.26 Settlement. Before or after a complaint under § 11.34 is filed, a settlement conference may occur between the OED...

  3. Simulation of LSI-11/PDP-11 series minicomputers

    International Nuclear Information System (INIS)

    Myers, J.R.; Cottrell, R.L.A.; Bricaud, B.M.

    1979-04-01

    A functional simulation of te PDP-11 series minicomputers was implemented to run either interactively or as a batch job on an IBM 370 computer. The simulator operates in two modes, the supervisor mode and the run mode. In the supervisor mode, the simulator implements a command language, which allows users to examine and change the contents of memory or other addressable registers in the simulated machine. Optionally, an instruction trace may also be turned on or off. In the run mode, the simulation of the instruction set is tested by successfully running DEC's MAINDEC basic instruction test on the simulated machine. The interrupt structure is modeled. The simulation is open ended in the sense that users may define new peripheral devices, by including their own FORTRAN callable subroutines for each simulated device. Currently the following devices are supported: floppy disks, a console terminal, a card reader, a card punch, a line printer, and a communications multiplexer (DH11). With these devices DEC's RT-11 versions 2C and 3B have been successfully run on the simulator. At SLAC this simulator is proving useful in debugging software for one-of-a-kind hardware configurations, such as communications front end processors, that are not readily accessible for stand alone testing. 5 figures

  4. Children's Fears: A Pre-9/11 and Post-9/11 Comparison Using the American Fear Survey Schedule for Children

    Science.gov (United States)

    Burnham, Joy J.

    2007-01-01

    Children are influenced by the salient events surrounding them (e.g., 9/11 terrorist attacks, Hurricane Katrina, massacre at Virginia Tech). In this study, the author examined fears of children and adolescents in Grades 2-12 in a pre-and post-September 11, 2001, comparison using the American Fear Survey Schedule for Children (FSSC-AM; J. J.…

  5. Beyond E 11

    Science.gov (United States)

    Bossard, Guillaume; Kleinschmidt, Axel; Palmkvist, Jakob; Pope, Christopher N.; Sezgin, Ergin

    2017-05-01

    We study the non-linear realisation of E 11 originally proposed by West with particular emphasis on the issue of linearised gauge invariance. Our analysis shows even at low levels that the conjectured equations can only be invariant under local gauge transformations if a certain section condition that has appeared in a different context in the E 11 literature is satisfied. This section condition also generalises the one known from exceptional field theory. Even with the section condition, the E 11 duality equation for gravity is known to miss the trace component of the spin connection. We propose an extended scheme based on an infinite-dimensional Lie superalgebra, called the tensor hierarchy algebra, that incorporates the section condition and resolves the above issue. The tensor hierarchy algebra defines a generalised differential complex, which provides a systematic description of gauge invariance and Bianchi identities. It furthermore provides an E 11 representation for the field strengths, for which we define a twisted first order self-duality equation underlying the dynamics.

  6. Production of ethanol from wet oxidised wheat straw by Thermoanaerobacter mathranii

    DEFF Research Database (Denmark)

    Ahring, B.K.; Licht, D.; Schmidt, A.S.

    1999-01-01

    evaluated with respect to total sugars, xylose, and 2-furfural produced. The concentration of sugars tended to be highest in hydrolysates produced at high oxygen pressures, whereas the concentration of 2-furfural was lowest in hydrolysates produced at low oxygen pressures and high carbonate concentrations...... with the commercial enzyme Celluclast(R) or with acid hydrolysis improved the ethanol yield from the hydrolysates. Treatment with Pentopan(TH) Mono BG or Pulpzyme(R) HC, both endo-1,4-beta-xylanases, had no effect neither had co-cultivation with the xylanase-producing Dictyoglomus B4. (C) 1998 Published by Elsevier...

  7. Combined enzymatic and physical deinking methodology for efficient eco-friendly recycling of old newsprint.

    Directory of Open Access Journals (Sweden)

    Antar Puneet Virk

    Full Text Available BACKGROUND: The development in the deinking process has made recycled fiber a major part of the raw material for pulp and paper industry. Enzymes have revolutionized the deinking process obtaining brightness levels surpassing conventional deinking processes. This study explores the deinking efficiencies of bacterial alkalophilic laccase (L and xylanase (X enzymes along with physical deinking methods of microwaving (MW and sonication (S for recycling of old newsprint (ONP. METHODS AND RESULTS: The operational parameters viz. enzyme dose, pH and treatment time for X and L deinking were optimized statistically using Response Surface Methodology. Laccase did not require any mediator supplementation for deinking. Deinking of ONP pulp with a combination of xylanase and laccase enzymes was investigated, and fiber surface composition and morphological changes were studied using X-ray diffraction, fourier transform infrared spectroscopy and scanning electron microscopy. Compared to the pulp deinked with xylanase (47.9% or laccase (62.2% individually, the percentage reduction of effective residual ink concentration (ERIC was higher for the combined xylanase/laccase-deinked pulp (65.8%. An increase in brightness (21.6%, breaking length (16.5%, burst factor (4.2% tear factor (6.9%, viscosity (13% and cellulose crystallinity (10.3% along with decrease in kappa number (22% and chemical consumption (50% were also observed. Surface appeared more fibrillar along with changes in surface functional groups. A combination of physical and enzymatic processes (S-MW-XL for deinking further improved brightness (28.8% and decreased ERIC (73.9% substantially. CONCLUSION: This is the first report on deinking of ONP with laccase without any mediator supplementation. XL pretreatment resulted in marked improvement in paper quality and a new sequence being reported for deinking (S-MW-XL will contribute further in decreasing chemical consumption and making the process

  8. Effect of peat extract on the hydrolytic enzymes of Phanerochaete chrysosporium

    Energy Technology Data Exchange (ETDEWEB)

    Nawaz, M; Gunasekaran, M

    1988-08-01

    Peat, a partially decomposed plant material rich in minerals and lignocellulose was tested as a substrate for the growth and production of hydrolytic enzymes viz. cellulase, cellobiase and xylanase in Phanerochaete chrysosporium. Three types of peat extracts such as cold, hot water and autoclaved were prepared and tested. Among them, autoclaved extract supported the maximal growth. The intracellular enzyme activities peaked on the fifth day after inoculation irrespective of the media and enzyme tested. Addition of cellobiose and lactose in the medium induced the production of cellulase, xylanase and to a lesser extent cellobiase. Addition of glucose and sucrose to the media resulted in the suppression of all the extracellular enzymes tested. 18 refs., 5 figs.

  9. Enzymatic pulp upgrade for producing high-value cellulose out of a Kraft paper pulp.

    Science.gov (United States)

    Hutterer, Christian; Kliba, Gerhard; Punz, Manuel; Fackler, Karin; Potthast, Antje

    2017-07-01

    The high-yield separation of polymeric parts from wood-derived lignocellulosic material is indispensable in biorefinery concepts. For the separation of cellulose and xylan from hardwood paper pulps to obtain pulps of high cellulose contents, simple alkaline extractions were found to be the most suitable technology, although having certain limitations. These are embodied by residual alkali resistant xylan incorporated in the pulp matrix. Further purification in order to produce pure cellulose with a low uniformity could be achieved selectively degrading residual xylan and depolymerizing the cellulose macromolecules by xylanase and cellulase. The latter help to adjust cellulose chain lengths for certain dissolving pulp grades while reducing the demand for ozone in subsequent TCF bleaching. Experiments applying different commercially available enzyme preparations revealed the dependency of xylanase performance on the residual xylan content in pulps being stimulated by additional cellulase usage. The action of the latter strongly depends on the cellulose allomorphy confirming the impact of the pulp morphology. Hence, the combined application of both types of enzymes offers a high potential for upgrading pulps in order to produce a pure and high-value cellulose product. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. 8.64-11.62 GHz CMOS VCO and divider in a zero-IF 802.11a/b/g WLAN and Bluetooth application

    International Nuclear Information System (INIS)

    Sun Yu; Mei Niansong; Lu Bo; Huang Yumei; Hong Zhiliang

    2010-01-01

    A fully integrated VCO and divider implemented in SMIC 0.13-μm RFCMOS 1P8M technology with a 1.2 V supply voltage is presented. The frequency of the VCO is tuning from 8.64 to 11.62 GHz while the quadrature LO signals for 802.11a WLAN in 5.8 GHz band or for 802.11b/g WLAN and Bluetooth in 2.4 GHz band can be obtained by a frequency division by 2 or 4, respectively. A 6 bit switched capacitor array is applied for precise tuning of all necessary frequency bands. The testing results show that the VCO has a phase noise of-113 dBc - 1 MHz offset from the carrier of 5.5 GHz by dividing VCO output by two and the VCO core consumes 3.72 mW. The figure-of-merit for the tuning-range (FOM T ) of the VCO is -192.6 dBc/Hz. (semiconductor integrated circuits)

  11. 8.64-11.62 GHz CMOS VCO and divider in a zero-IF 802.11a/b/g WLAN and Bluetooth application

    Energy Technology Data Exchange (ETDEWEB)

    Sun Yu; Mei Niansong; Lu Bo; Huang Yumei; Hong Zhiliang, E-mail: yumeihuang@fudan.edu.c [ASIC and System State Key Laboratory, Fudan University, Shanghai 201203 (China)

    2010-10-15

    A fully integrated VCO and divider implemented in SMIC 0.13-{mu}m RFCMOS 1P8M technology with a 1.2 V supply voltage is presented. The frequency of the VCO is tuning from 8.64 to 11.62 GHz while the quadrature LO signals for 802.11a WLAN in 5.8 GHz band or for 802.11b/g WLAN and Bluetooth in 2.4 GHz band can be obtained by a frequency division by 2 or 4, respectively. A 6 bit switched capacitor array is applied for precise tuning of all necessary frequency bands. The testing results show that the VCO has a phase noise of-113 dBc - 1 MHz offset from the carrier of 5.5 GHz by dividing VCO output by two and the VCO core consumes 3.72 mW. The figure-of-merit for the tuning-range (FOM{sub T}) of the VCO is -192.6 dBc/Hz. (semiconductor integrated circuits)

  12. Synthesis of [11C]-Sarin

    International Nuclear Information System (INIS)

    Prenant, C.; Crouzel, C.

    1990-01-01

    [ 11 C]-acetone, prepared from 11 CO 2 and methyllithium, was reduced to [ 11 C]-isopropanol. The latter reacted with methylphosphonic acid difluoride in the presence of diisopropylamine, to yield [11C]-sarin. 3.4 GBq (100 mCi) of [ 11 C]-sarin may be obtained from about 55.5 GBq (1.5 Ci) of 11 CO 2 in 40 minutes. The product purified by HPLC, is obtained with a specific radioactivity ranging from 22.2 to 33.3 GBq (600 to 900 mCi/μmol.). (author)

  13. WOOD 1:1

    DEFF Research Database (Denmark)

    2013-01-01

    Report from 1:1 workshop at Virserum Konsthall in May 2013 with a team of Master's students from Royal Academy of Fine Arts, School of Architecture, Dpt. 3.......Report from 1:1 workshop at Virserum Konsthall in May 2013 with a team of Master's students from Royal Academy of Fine Arts, School of Architecture, Dpt. 3....

  14. Solute carrier protein family 11 member 1 (Slc11a1) activation efficiently inhibits Leishmania donovani survival in host macrophages.

    Science.gov (United States)

    Singh, Nisha; Gedda, Mallikarjuna Rao; Tiwari, Neeraj; Singh, Suya P; Bajpai, Surabhi; Singh, Rakesh K

    2017-09-01

    Visceral leishmaniasis (kala-azar), a life threatening disease caused by L. donovani , is a latent threat to more than 147 million people living in disease endemic South East Asia region of the Indian subcontinent. The therapeutic option to control leishmanial infections are very limited, and at present comprise only two drugs, an antifungal amphotericin B and an antitumor miltefosine, which are also highly vulnerable for parasitic resistance. Therefore, identification and development of alternate control measures is an exigent requirement to control leishmanial infections. In this study, we report that functionally induced expression of solute carrier protein family 11 member 1 ( Slc11a1), a transmembrane divalent cationic transporter recruited on the surface of phagolysosomes after phagocytosis of parasites, effectively inhibits Leishmania donovani growth in host macrophages. Further, the increased Slc11a1 functionality also resulted in increased production of NOx, TNF-α and IL-12 by activated macrophages. The findings of this study signify the importance of interplay between Slc11a1 expression and macrophages activation that can be effectively used to control of Leishmania growth and survival.

  15. Piloting a Collaborative Web-Based System for Testing ICD-11.

    Science.gov (United States)

    Donada, Marc; Kostanjsek, Nenad; Della Mea, Vincenzo; Celik, Can; Jakob, Robert

    2017-01-01

    The 11th revision of the International Classification of Diseases (ICD-11), for the first time in ICD history, deployed web-based collaboration of experts and ICT tools. To ensure that ICD-11 is working well, it needs to be systematically field tested in different settings, across the world. This will be done by means of a number of experiments. In order to support its implementation, a web-based system (ICDfit) has been designed and developed. The present paper illustrates the current prototype of the system and its technical testing. the system has been designed according to WHO requirements, and implemented using PHP and MySQL. Then, a preliminary technical test has been designed and run in January 2016, involving 8 users. They had to carry out double coding, that is, coding case summaries with both ICD-10 and ICD-11, and answering quick questions on the coding difficulty. the 8 users coded 632 cases each, spending an average of 163 seconds per case. While we found an issue in the mechanism used to record coding times, no further issues were found. the proposed system seems to be technically adequate for supporting future ICD-11 testing.

  16. CD5−NK1.1+ γδ T Cells that Develop in a Bcl11b-Independent Manner Participate in Early Protection against Infection

    Directory of Open Access Journals (Sweden)

    Shinya Hatano

    2017-10-01

    Full Text Availabl