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Sample records for chaetomium thermophilum xylanase

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

    International Nuclear Information System (INIS)

    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)

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

    International Nuclear Information System (INIS)

    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)

  3. 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...... xylanase treatment and increased to 34 % when xylanase was combined with rCtAxeA and rCtAxeB. In sum, the present study first report the biochemical characterization of two acetyl xylan esterases from C. thermophilum, which are efficient in hydrolyzing hemicellulose with potential application in biomass...

  4. Developing genetic tools to exploit Chaetomium thermophilum for biochemical analyses of eukaryotic macromolecular assemblies

    OpenAIRE

    Nikola Kellner; Johannes Schwarz; Miriam Sturm; Javier Fernandez-Martinez; Sabine Griesel; Wenzhu Zhang; Chait, Brian T.; Rout, Michael P.; Ulrich Kück; Ed Hurt

    2016-01-01

    We describe a method to genetically manipulate Chaetomium thermophilum, a eukaryotic thermophile, along with various biochemical applications. The transformation method depends on a thermostable endogenous selection marker operating at high temperatures combined with chromosomal integration of target genes. Our technique allows exploiting eukaryotic thermophiles as source for purifying thermostable native macromolecular complexes with an emphasis on the nuclear pore complex, holding great pot...

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

    International Nuclear Information System (INIS)

    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

  6. 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.

  7. Chaetomium

    Czech Academy of Sciences Publication Activity Database

    Hubka, Vít

    Boca Raton: CRC Press, 2015, s. 211-228. ISBN 978-1-4665-5986-8 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61388971 Keywords : chaetomium * infection Subject RIV: EC - Immunology

  8. 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 stability at extended acidic-alkaline pH region, and noticeable thermostability. CtFae released ferulic acid from wheat insoluble arabinoxylan, as well as ferulic and p-coumaric acids from wheat straw and ryegrass, indicating potentials for industrial applications like biomass conversion in...

  9. Ecology of Thermophilic Fungi in Mushroom Compost, with Emphasis on Scytalidium thermophilum and Growth Stimulation of Agaricus bisporus Mycelium

    OpenAIRE

    Straatsma, Gerben; Samson, Robert A.; Olijnsma, Tineke W.; Op den Camp, Huub J. M.; Gerrits, Jan P. G.; Van Griensven, Leo J. L. D.

    1994-01-01

    Twenty-two species of thermophilic fungi were isolated from mushroom compost. Scytalidium thermophilum was present in the compost ingredients, fresh straw, horse droppings, and drainage from compost and dominated the fungal biota of compost after preparation. Of 34 species of thermophilic fungi tested, 9 promoted mycelial growth of Agaricus bisporus on sterilized compost: Chaetomium thermophilum, an unidentified Chaetomium sp., Malbranchea sulfurea, Myriococcum thermophilum, S. thermophilum, ...

  10. Identification and characterization of diverse xylanases from thermophilic and thermotolerant fungi

    Directory of Open Access Journals (Sweden)

    Bhat, M. K.

    2006-07-01

    Full Text Available Thirteen fungal isolates included in this study expressed multiple xylanase isoforms as observed by xylan zymograms of polyacrylamide gel electrophoresis (PAGE and isoelectrofocussing (IEF fractionated proteins. Eighty-three xylanases produced by these thermophilic and thermotolerant strains were detected using the IEF profiling technique. Xylanases identified on the basis of their isoelectric points (pI were functionally diverse and exhibited differential catalytic activities against various xylan types (birch wood xylan, larch wood xylan, oat spelt xylan, rye arabino xylan and wheat arabino xylan as well as debranched arabinan. Thermophilic isolates, Chaetomium thermophilum, Humicola insolens, Melanocarpus sp., Malbranchea sp. and Thermoascus aurantiacus, were found to produce alkaline active xylanases that showed a bleach boosting effect on Decker pulp resulting in increased brightness (1.60-2.04 ISO units.

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

    DEFF Research Database (Denmark)

    Borkhardt, Bernhard; Harholt, Jesper; Ulvskov, Peter Bjarne; Ahring, Birgitte Kiær; Jørgensen, Bodil; Brinch-Pedersen, Henrik

    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. Tra...

  12. Molecular cloning and characterization of a GH11 endoxylanase from Chaetomium globosum, and its use in enzymatic pretreatment of biomass

    DEFF Research Database (Denmark)

    Singh, Raushan Kumar; Tiwari, Manish Kumar; Kim, Dongwook;

    2013-01-01

    An endo-1,4-β-xylanase gene, xylcg, was cloned from Chaetomium globosum and successfully expressed in Escherichia coli. The complete gene of 675 bp was amplified, cloned into the pET 28(a) vector, and expressed. The optimal conditions for the highest activity of the purified recombinant XylCg were......Cg belongs to the GH11 family. Rice straw pretreated with XylCg showed 30 % higher conversion yield than the rice straw pretreated with a commercial xylanase. Although xylanases have been characterized from fungal and bacterial sources, C. globosum XylCg is distinguished from other xylanases by its high...

  13. Probing the role of sigma π interaction and energetics in the catalytic efficiency of endo-1,4-β-xylanase

    DEFF Research Database (Denmark)

    Singh, Raushan Kumar; Tiwari, Manish Kumar; Kim, In-Won;

    2012-01-01

    Chaetomium globosum endo-1,4-β-xylanase (XylCg) is distinguished from other xylanases by its high turnover rate (1,860 s(-1)), the highest ever reported for fungal xylanases. One conserved amino acid, W48, in the substrate binding pocket of wild-type XylCg was identified as an important residue...

  14. Controle biológico de Monosporascus cannonballus com Chaetomium Biological control of Monosporascus cannonballus by Chaetomium

    Directory of Open Access Journals (Sweden)

    Rui S. Júnior

    2007-02-01

    Full Text Available O colapso do meloeiro (Cucumis melo, causado por Monosporascus cannonballus, é uma das principais enfermidades que acometem esta olerícola. O presente trabalho objetivou estudar o potencial do controle biológico de M. cannonballus com Chaetomium. Substrato infestado com 0,5; 1; 2; 4 e 8 x 10(5 UFC g-1 de Chaetomium foi colocado em bandejas, nas quais sementes de meloeiro do tipo Pele de Sapo cv. PS 1430 foram semeadas após duas semanas de incubação. As mudas obtidas foram transplantadas para vasos com substrato semelhante, sendo que neste momento o substrato foi infestado com mais 2,5 x 10(4 UFC g-1 de Chaetomium por vaso e 20 UFC g-1 de substrato de M. cannonballus. Três testemunhas foram utilizadas, uma infestada apenas com Chaetomium, outra somente com M. cannonballus e testemunha absoluta. O delineamento estatístico foi inteiramente casualizado com cinco repetições. As avaliações foram efetuadas aos 45 dias após o transplante, sendo calculado o índice geral de doença e o percentual de controle. O valor do índice geral de doença variou conforme o aumento da concentração de Chaetomium, sendo os menores valores encontrados para os tratamentos 4 e 8 x 10(5 UFC g-1de Chaetomium equivalentes a 0,9 e 0,6, respectivamente. A análise estatística demonstrou que os tratamentos correspondentes a 4 e 8 x 10(5 UFC g-1 de Chaetomium foram mais eficientes, diferindo dos demais, com percentual de controle superior a 55 %, evidenciado o potencial de Chaetomium no controle de M. cannonballus.Melon collapse caused by Monosporascus cannonballus is one of the main diseases that affect this cucurbit. The objective of this research was to study the biological control of M. cannonballus by Chaetomium. Infested substrates with 0.5, 1, 2, 4 and 8 x 10(5 CFU g-1 were introduced in plates where seeds of melon type Piel de Sapo cv. PS 1430 were sowed in seeding trays after two weeks of incubation. The seedlings were transplanted to pots with a

  15. 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...

  16. First Spanish case of onychomycosis caused by Chaetomium globosum.

    Science.gov (United States)

    Aspiroz, Carmen; Gené, Josepa; Rezusta, Antonio; Charlez, Luis; Summerbell, Richard C

    2007-05-01

    Members of the fungal genus Chaetomium usually colonize cellulose-containing plant remains but on rare occasions may cause opportunistic mycoses and cutaneous infection in otherwise healthy individuals. To our knowledge, there have been only five credible descriptions of onychomycosis caused by members of this genus and only two of these contained information on therapy. We describe the first case of Chaetomium globosum onychomycosis recorded in Spain. The etiologic significance of the fungus was confirmed by its repeated isolation at different times, to the exclusion of dermatophytes. Clinically, the affected nails showed an excellent response to terbinafine and complete cure appeared to have been attained. PMID:17464849

  17. Controle biológico de Monosporascus cannonballus com Chaetomium Biological control of Monosporascus cannonballus by Chaetomium

    OpenAIRE

    Rui S. Júnior; Roberto Beltrán; Antonio Vicent; Josep Armengol; José García-Jiménez; Érika V. Medeiros

    2007-01-01

    O colapso do meloeiro (Cucumis melo), causado por Monosporascus cannonballus, é uma das principais enfermidades que acometem esta olerícola. O presente trabalho objetivou estudar o potencial do controle biológico de M. cannonballus com Chaetomium. Substrato infestado com 0,5; 1; 2; 4 e 8 x 10(5) UFC g-1 de Chaetomium foi colocado em bandejas, nas quais sementes de meloeiro do tipo Pele de Sapo cv. PS 1430 foram semeadas após duas semanas de incubação. As mudas obtidas foram transplantadas par...

  18. Implantation phaeohyphomycosis caused by a non-sporulating Chaetomium species.

    Science.gov (United States)

    Najafzadeh, M J; Fata, A; Naseri, A; Keisari, M Saradeghi; Farahyar, S; Ganjbakhsh, M; Ziaee, M; Dolatabadi, S; de Hoog, G S

    2014-06-01

    We report the case of a 66-year-old Iranian woman with a phaeohyphomycotic cyst (approximately 3×2.5cm in size) on the right lateral side of the neck. She had dysphagia and hoarseness, without any pain. She complained about discharge of black liquid on the skin and irritation. Histological examination of biopsy fragments from the lesions showed septate, branched brown hyphae. The fungus was cultured, but sporulation remained absent from 4- week-old cultures on Sabouraud dextrose agar (SDA), malt extract agar (MEA), potato dextrose agar (PDA), and water agar with sterile filter paper. Identification with the genus Chaetomium was achieved by sequencing the internal transcribed spacer (ITS) and the small subunit (SSU) domains of the rDNA gene and comparison with sequences held at GenBank and at the Centraalbureau voor Schimmelcultures (CBS). Sequencing of the SSU rRNA gene reveals this strain as belonging to the genus Chaetomium. The sequence of ITS did not fully match with any sequence of available ex-type strains of Chaetomium, Thielavia, Madurella and Papulaspora and hence might belong to an undescribed species. However, without diagnostic morphological features the taxon cannot be introduced as a novel member of the genus Chaetomium. After local excision of the cyst and antifungal therapy with ketoconazole (200mg twice a day), the lesion regressed and healed completely. PMID:24246716

  19. Pangola grass colonized with Scytalidium thermophilum for production of Agaricus bisporus.

    Science.gov (United States)

    Sanchez, Jose E; Mejia, Laura; Royse, Daniel J

    2008-02-01

    This work had the dual objective of selecting a substrate for rapid mycelial growth of Scytalidium thermophilum and then comparing the growth and production of a brown variety of Agaricus bisporus on substrate non-colonized and colonized with S. thermophilum. Mycelial growth of S. thermophilum at 45 degrees C was significantly greater on potato dextrose yeast extract agar (0.58 mm/h) as compared to malt extract glucose agar (0.24 mm/h) and yeast extract glucose agar (0.44 mm/h). On cereal grain, S. thermophilum grew significantly faster on rice (0.31 mm/h) compared to sorghum (0.22 mm/h) and millet (0.18 mm/h). It also grew faster on Pangola grass (0.49 mm/h) compared to corncobs (0.30 mm/h) and sawdust (0.18 mm/h). Colonization of Pangola grass with S. thermophilum was influenced by the addition of calcium salts in the form of gypsum, hydrated lime and ground limestone. For production of A. bisporus, biological efficiency (BE) on pasteurized Pangola grass pre-colonized by S. thermophilum for 4 days at 45 degrees C was more than twice (26.4%) that on grass non-colonized by S. thermophilum (11.0%). The addition of 2% hydrated lime to Pangola grass prior to colonization by S. thermophilum resulted in an additional doubling of BE of mushroom production (48.1%). These results show the possibility of developing a non-composted substrate method for producing A. bisporus without autoclaving the substrate. PMID:17331714

  20. Endophytic Chaetomium globosum enhances maize seedling copper stress tolerance.

    Science.gov (United States)

    Abou Alhamed, M F; Shebany, Y M

    2012-09-01

    This study aims at characterisation of the impact of Chaetomium globosum on copper stress resistance of maize seedlings. Higher levels of copper treatment decreased maize dry weight and induced a marked increase in osmotic solutes, antioxidant enzyme activity and the level of lipid peroxidation. On the other hand, addition of the endophytic C. globosum alleviated the toxic effect of copper on maize growth. The combination of copper sulphate and Chaetomium increased seedling dry weight, osmotic solute content and antioxidant enzyme activity compared to copper sulphate alone, while lipid peroxidation levels were also decreased. The fungal scavenger system might be important for supporting the ability of maize seedlings to resist copper toxicity. PMID:22672065

  1. FERROFLUIDS INFLUENCE ON DEHYDROGENASES ACTIVITY IN CELLULOLYTIC FUNGUS CHAETOMIUM GLOBOSUM

    OpenAIRE

    Alexandru Manoliu; Lacramioara Oprica; Zenovia Olteanu; Dorina Creanga

    2003-01-01

    he activity of dehy drogenases was studied after ferrofluids supply ing in the culture medium of Chaetomium globosum. Spectral measurements were carried out after 7 and, respectively , 11 day s of growth. Different results were noticed for different ferrofluids concentrations: 20, 40, 60, 80 and 100 μl/L. Inhibitory or stimulatory ferrofluids effect was obtained depending on the nature of the investigated enzyme.

  2. Chaetomium and Stachybotrys in water-damaged buildings

    DEFF Research Database (Denmark)

    Andersen, Birgitte; Lewinska, Anna Malgorzata; Nielsen, Jakob Blæsbjerg; Dosen, Ina; Nielsen, Kristian Fog; Peuhkuri, Ruut Hannele; Rode, Carsten; Clausen, Geo; Thrane, Ulf

    Fungal growth occurs when parts of the building envelope get very wet due to unfortunate combinations of factors, e.g. thermal bridges/lack of ventilation, shoddy foundations/flooding or leaks in build-in pipes. Chaetomium and Stachybotrys are not as abundant as Penicillium and Aspergillus (Table......), however, they may produce volatiles and microparticles that can cause health problems. They are common in wet walls constructed of wood fibre board (OSB/plywood) and gypsum board....

  3. Nutrient requirements and growth physiology of the photoheterotrophic Acidobacterium, Chloracidobacterium thermophilum

    Directory of Open Access Journals (Sweden)

    Donald A Bryant

    2015-03-01

    Full Text Available A novel thermophilic, microaerophilic, anoxygenic and chlorophototrophic member of the phylum Acidobacteria, Chloracidobacterium thermophilum strain BT, was isolated from a cyanobacterial enrichment culture derived from microbial mats associated with Octopus Spring, Yellowstone National Park, Wyoming. C. thermophilum is strictly dependent on light and oxygen and grows optimally as a photoheterotroph at irradiance values between 20 to 50 µmol photons m-2 s-1. C. thermophilum is unable to synthesize branched-chain amino acids, L-lysine, and vitamin B12, which are required for growth. Although the organism lacks genes for autotrophic carbon fixation, bicarbonate is also required. Mixtures of other amino acids and 2-oxoglutarate stiumulate growth. As suggested from genomic sequence data, C. thermophilum requires a reduced sulfur source such as thioglycolate, cysteine, methionine, or thiosulfate. The organism can be grown in a defined medium at 51°C (Topt; range 44 to 58°C in the pH range 5.5 to 9.5 (pHopt = ~7.0. Using the defined growth medium and optimal conditions, it was possible to isolate new C. thermophilum strains directly from samples of hot spring mats Yellowstone National Park, Wyoming. The new isolates differ from the type strain with respect to pigment composition, morphology in liquid culture, and temperature adaptation.

  4. Genome sequence of Symbiobacterium thermophilum, an uncultivable bacterium that depends on microbial commensalism

    OpenAIRE

    Ueda, Kenji; YAMASHITA Atsushi; Ishikawa, Jun; Shimada, Masafumi; Watsuji, Tomo-o; Morimura, Kohji; Ikeda, Haruo; Hattori, Masahira; Beppu, Teruhiko

    2004-01-01

    Symbiobacterium thermophilum is an uncultivable bacterium isolated from compost that depends on microbial commensalism. The 16S ribosomal DNA-based phylogeny suggests that this bacterium belongs to an unknown taxon in the Gram-positive bacterial cluster. Here, we describe the 3.57 Mb genome sequence of S.thermophilum. The genome consists of 3338 protein-coding sequences, out of which 2082 have functional assignments. Despite the high G + C content (68.7%), the genome is closest to that of Fir...

  5. 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...

  6. A case report of a mixed Chaetomium globosum/Trichophyton mentagrophytes onychomycosis

    OpenAIRE

    Lagacé, Jean; Cellier, Eric

    2012-01-01

    Recently, an increasing prevalence of nondermatophyte mold onychomycosis was observed, in which Chaetomium globosum was rarely involved as primary pathogenic agent. Besides this, reports of mixed infection associating a dermatophyte and a nondermatophyte mold have become more frequent. Here, we present a clinical case of a mixed onychomycosis infection of a toenail caused by Chaetomium globosum and Trichophyton mentagrophytes. To our knowledge, this specific association is reported for the fi...

  7. [Progress in the thermophilic and alkalophilic xylanases].

    Science.gov (United States)

    Bai, Wenqin; Wang, Qinhong; Ma, Yanhe

    2014-06-01

    Xylanase is the key enzyme to degrade xylan that is a major component of hemicellulose. The enzyme has potential industrial applications in the food, feed, paper and flax degumming industries. The use of xylanases becomes more and more important in the paper industry for bleaching purposes. Xylanases used in the pulp bleaching process should be stable and active at high temperature and alkaline pH. Thermophilic and alkalophilic xylanases could be obtained by screening the wild type xylanases or engineering the mesophilic and neutral enzymes. In this paper, we reviewed recent progress of screening of the thermophilic and alkalophilic xylanases, molecular mechanism of thermal and alkaline adaptation and molecular engineering. Future research prospective was also discussed. PMID:25212001

  8. Comparative characterization of commercially important xylanase enzymes

    OpenAIRE

    Arora, Neelima; Banerjee, Amit Kumar; Mutyala, Srilaxmi; Murty, Upadhyayula Suryanarayana

    2009-01-01

    Xylanase is an industrially important enzyme having wide range of applications especially in paper industry. It is crucial to gain an understanding about the structure and functional aspects of various xylanases produced from diverse sources. In this study, a bioinformatics and molecular modeling approach was adopted to explore properties and structure of xylanases. Physico-chemical properties were predicted and prediction of motifs, disulfide bridges and secondary structure was performed for...

  9. Xylanase inhibitors bind to nonstarch polysaccharides.

    Science.gov (United States)

    Fierens, Ellen; Gebruers, Kurt; Courtin, Christophe M; Delcour, Jan A

    2008-01-23

    This study is an in-depth investigation of the interaction between polysaccharides and the proteinaceous xylanase inhibitors, Triticum aestivum xylanase inhibitor (TAXI), xylanase inhibitor protein (XIP), and thaumatin-like xylanase inhibitor (TLXI). The binding affinities of all three known types of xylanase inhibitors from wheat are studied by measuring the residual xylanase inhibition activity after incubation of the inhibitors in the presence of different polysaccharides, such as beta-glucans and (arabino)xylans. The binding affinities of all three xylanase inhibitors for (arabino)xylans increased with a decreasing arabinose/xylose ratio (A/X ratio). This phenomenon was observed both with water-extractable and water-unextractable (arabino)xylans. The inhibitors also interacted with different soluble and insoluble beta-glucans. None of the inhibitors tested had the ability to hydrolyze the polysaccharides investigated. The present findings contribute to the unraveling of the function of xylanase inhibitors in nature and to the prediction of the effect of added xylanases in cereal-based biotechnological processes, such as bread making and gluten-starch separation. PMID:18092758

  10. [Chemical constituents from endophyte Chaetomium globosum in Imperata cylindrical].

    Science.gov (United States)

    Shen, Li; Zhu, Li; Wei, Zhong-qi; Li, Xiao-wen; Li, Ming; Song, Yong-chun

    2015-12-01

    Isolation and purification of chemical constituents from solid culture of endophyte Chaetomium globosum in Imperata cylindrical was performed through silica gel column chromatography, gel filtration over Sephadex LH-20 and preparative HPLC. Nine compounds were obtained and their structures were determined as chaetoglobosin F(1), chaetoglobosin Fex(2), chaetoglobosin E(3) cytoglobosin A(4), penochalasin C(S), isochaetoglobosin D (6), N-benzoylphenylalaninyl-N-benzoyphenylalaninate(7), uracil(8) and 5-methyluracil(9), respectively, based on HR-MS and NMR data and comparison with literatures. Compound 7 was isolated from Chaeeomium sp. for the first time. In vitro cytotoxicity of compounds was evaluated using MTT mothed and 1,3,4 and 5 showed inhibition activity to the human cervical carcinoma cell HeLa with IC50 values of 99.43, 23.77, 97.92, 86.25 micromol x L(-1), while positive cotolocisnin Ad apno1ch alse IC50 24.33 micromol x L(-1). PMID:27141677

  11. INDUSTRIAL APPLICATIONS AND FUTURE PROSPECTS OF MICROBIAL XYLANASES: A REVIEW

    OpenAIRE

    Saurabh Sudha Dhiman; Jitender Sharma; Bindu Battan

    2008-01-01

    Microbial enzymes such as xylanases enable new technologies for industrial processes. Xylanases (xylanolytic enzyme) hydrolyze complex polysaccharides like xylan. Research during the past few decades has been dedicated to enhanced production, purification, and characterization of microbial xylanase. But for commercial applications detailed knowledge of regulatory mechanisms governing enzyme production and functioning should be required. Since application of xylanase in the commercial sector i...

  12. PCR and real-time PCR primers developed for detection and identification of Bifidobacterium thermophilum in faeces

    Directory of Open Access Journals (Sweden)

    Mini Raffaella

    2008-10-01

    Full Text Available Abstract Background Culture-independent methods based on the 16S ribosomal RNA molecule are nowadays widely used for assessment of the composition of the intestinal microbiota, in relation to host health or probiotic efficacy. Because Bifidobacterium thermophilum was only recently isolated from human faeces until now, no specific real-time PCR (qPCR assay has been developed for detection of this species as component of the bifidobacterial community of the human intestinal flora. Results Design of specific primers and probe was achieved based on comparison of 108 published bifidobacterial 16S rDNA sequences with the recently published sequence of the human faecal isolate B. thermophilum RBL67. Specificity of the primer was tested in silico by similarity search against the sequence database and confirmed experimentally by PCR amplification on 17 Bifidobacterium strains, representing 12 different species, and two Lactobacillus strains. The qPCR assay developed was linear for B. thermophilum RBL67 DNA quantities ranging from 0.02 ng/μl to 200 ng/μl and showed a detection limit of 105 cells per gram faeces. The application of this new qPCR assay allowed to detect the presence of B. thermophilum in one sample from a 6-month old breast-fed baby among 17 human faecal samples tested. Additionally, the specific qPCR primers in combination with selective plating experiments led to the isolation of F9K9, a faecal isolate from a 4-month old breast-fed baby. The 16S rDNA sequence of this isolate is 99.93% similar to that of B. thermophilum RBL67 and confirmed the applicability of the new qPCR assay in faecal samples. Conclusion A new B. thermophilum-specific qPCR assay was developed based on species-specific target nucleotides in the 16S rDNA. It can be used to further characterize the composition of the bifidobacterial community in the human gastrointestinal tract. Until recently, B. thermophilum was considered as a species of animal origin, but here we

  13. Draft Genome Sequence of the Photoheterotrophic Chloracidobacterium thermophilum Strain OC1 Found in a Mat at Ojo Caliente

    OpenAIRE

    Hallenbeck, Patrick C.; Grogger, Melanie; Mraz, Megan; Veverka, Donald

    2016-01-01

    Metagenomics of an enrichment culture from a New Mexico hot spring allowed the description of a draft genome of a Chloracidobacterium thermophilum strain for the first time outside Yellowstone National Park with a surprisingly high degree of identity with the type strain.

  14. Biocontrol potential of Steinernema thermophilum and its symbiont Xenorhabdus indica against lepidopteran pests: virulence to egg and larval stages

    Science.gov (United States)

    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 ...

  15. Optimization and characterization of thermostable β-glucosidase by Scytalidium thermophilum SKESMBKU02

    Directory of Open Access Journals (Sweden)

    S. Santosh Kumar

    2015-06-01

    Full Text Available Scytalidium thermophilum SKESMBKU02, a thermophilic fungus isolated from cattle dung, produced a significant amount of thermostable β-glucosidase in Mandel and Weber medium (45°C, 3 days. Scytalidium thermophilum SKESMBKU02 opted pH – 5, temperature of 45°C for production of β-glucosidases. The glucose and yeast extract are the best carbon and nitrogen sources for β-glucosidase production. It has found that shake flak culture method exerted more β-glucosidase production than the still culture method. The fungus showed maximum dry weight in cellulose as carbon sources, yeast extract and malt extract as nitrogen source, pH of 9.0-10.0 and temperature of 45oC.The crude enzyme was characterized and found that substrate concentration of 6 mM and enzyme concentration of 300 μl are found suitable for the maximum β-glucosidase activity. Stability studies reveals that the enzyme was active at wide range of pH and temperature.

  16. Characterization of cellulase enzyme produced by Chaetomium sp. isolated from books and archives

    Directory of Open Access Journals (Sweden)

    Moza Mohammed AL-Kharousi

    2015-12-01

    Full Text Available Background: Cellulase is an important industrial enzyme used to degrade cellulosic biomass. The demand for cellulase enzyme is continuously increasing because of its applications in various industries. Hence, screening of cellulase producing microorganisms from different sources has gained significant importance. Material and Methods: In this study, fungi isolated from books and archives were screened for their cellulase producing abilities. Four different fungi were isolated from books and archives using potato dextrose agar. Screening of these isolates for cellulase production was carried out using carboxymethyl cellulose broth. The most efficient fungus was subjected to cellulase fermentation and enzymes produced were purified and partially characterized. Results: Four different fungi, Chaetomium sp., Aspergillus niger, Aspergillus nidulans and Penicillium sp., were isolated from books and archives. All the isolates were tested for their ability to producecellulase enzyme. During the primary screening Chaetomium sp. showed good growth and highercellulase activity (155.3±25.6 U/mL in carboxymethyl cellulose medium than the other fungi. The cellulase fermentation study was conducted with Chaetomium sp. using carboxymethyl cellulose asa substrate. During the stationary phase (144 h of the growth, the cellulase activity of Chaetomium sp. was significantly high. The maximum mycelial weight of this fungi was obtained at 168 h. Viscosity of the Chaetomium sp. inoculated fermentation medium continuously decreased until 144 h because of the degradation of carboxymethyl cellulose. During cellulase fermentation, pHincreased from the initial neutral pH to 8.5. Purified cellulase showed a specific activity of 7.3 U/mg. It exhibited maximum activity at 20°C and was stable between pH 5 and 9. Conclusions: Books and archives could be a good source for the isolation of cellulase producing fungi.

  17. Structural model and spectroscopic characteristics of the FMO antenna protein from the aerobic chlorophototroph, Candidatus Chloracidobacterium thermophilum

    Energy Technology Data Exchange (ETDEWEB)

    Wen, Jianzhong; Tsukatani, Yusuke; Cui, Weidong; Zhang, Hao; Gross, Michael L; Bryant, Donald A; Blankenship, R. E.

    The Fenna–Matthews–Olson protein (FMO) binds seven or eight bacteriochlorophyll a (BChl a) molecules and is an important model antenna system for understanding pigment-protein interactions and mechanistic aspects of photosynthetic light harvesting. FMO proteins of green sulfur bacteria (Chlorobiales) have been extensively studied using a wide range of spectroscopic and theoretical approaches because of their stability, the spectral resolution of their pigments, their water-soluble nature, and the availability of high-resolution structural data. We obtained new structural and spectroscopic insights by studying the FMO protein from the recently discovered, aerobic phototrophic acidobacterium, Candidatus Chloracidobacterium thermophilum. Native C. thermophilum FMO is a trimer according to both analytical gel filtration and native-electrospray mass spectrometry. Furthermore, the mass of intact FMO trimer is consistent with the presence of 21–24 BChl a in each. Homology modeling of the C. thermophilum FMO was performed by using the structure of the FMO protein from Chlorobaculum tepidum as a template. C. thermophilum FMO differs from C. tepidum FMO in two distinct regions: the baseplate, CsmA-binding region and a region that is proposed to bind the reaction center subunit, PscA. C. thermophilum FMO has two fluorescence emission peaks at room temperature but only one at 77 K. Temperature-dependent fluorescence spectroscopy showed that the two room-temperature emission peaks result from two excited-state BChl a populations that have identical fluorescence lifetimes. Modeling of the data suggests that the two populations contain 1–2 BChl and 5–6 BChl a molecules and that thermal equilibrium effects modulate the relative population of the two emitting states.

  18. Partial purification and characterization of xylanase produced by Penicillium expansum

    OpenAIRE

    André Luiz de Souza Querido; Jorge Luiz Cavalcante Coelho; Elza Fernandes Araújo; Virgínia Maria Chaves-Alves

    2006-01-01

    An extracellular xylanase was found to be the major protein in the filtrate culture of Penicillium expansum when grown on 0.3 % wheat bran, which showed no xylanase multiplicity. The enzime was partial purified by.ammonium sulfate fractioning, molecular exclusion chromatography, ultrafiltration and anion exchange chromatography. The protein eluation profile showed only one form of xylanase that was partially characterized. The activity of purified xylanase was optimal at pH 5.5 and 40 ºC. The...

  19. Bioactive secondary metabolites from the endophytic fungus Chaetomium sp. isolated from Salvia officinalis growing in Morocco

    Directory of Open Access Journals (Sweden)

    Ebel R.

    2009-01-01

    Full Text Available This study reports the chemical investigation and cytotoxic activity of the secondary metabolites produced by the endophytic fungus Chaetomium sp. isolated from Salvia officinalis growing in Morocco. This plant was collected from the Beni-Mellal Mountain in Morocco and belongs to the Lamiaceae family and is named in Morocco “Salmia”. The endophytic fungus Chaetomium sp. was isolated from the tissues of the stem of this plant. The fungal strain was identified by PCR. The crude organic extract of the fungal strain was proven to be active when tested for cytotoxicity against L5178Y mouse lymphoma cells. Chemical investigation of the secondary metabolites showed that cochliodinol is the main component beside isocochliodinol. The structures of the isolated compounds were determined on the basis of NMR analysis (1H, 13C, COSY and HMBC as well as by mass spectrometry using ESI (Electron Spray Ionisation as source.

  20. Functional Analysis of the Degradation of Cellulosic Substrates by a Chaetomium globosum Endophytic Isolate

    OpenAIRE

    Longoni, Paolo; Rodolfi, Marinella; Pantaleoni, Laura; Doria, Enrico; Concia, Lorenzo; Picco, Anna Maria; Cella, Rino

    2012-01-01

    Most photosynthetically fixed carbon is contained in cell wall polymers present in plant biomasses, the largest organic carbon source in the biosphere. The degradation of these polymers for biotechnological purposes requires the combined action of several enzymes. To identify new activities, we examined which enzymes are activated by an endophytic strain of Chaetomium globosum to degrade cellulose-containing substrates. After biochemical analyses of the secretome of the fungus grown on cellul...

  1. Antifungal Metabolites Produced by Chaetomium globosum No.04, an Endophytic Fungus Isolated from Ginkgo biloba

    OpenAIRE

    Zhang, Guizhen; Zhang, Yanhua; Qin, Jianchun; Qu, Xiaoyan; Liu, Jinliang; Xiang LI; Pan, Hongyu

    2013-01-01

    The fungal endophyte Chaetomium globosum No.04 was isolated from the medicinal plant Ginkgo biloba. The crude extract of the fungus fermentation were active in the agar-diffusion tests against the phytopathogenic fungi Rhizopus stolonifer and Coniothyrium diplodiella. Further bioassay-guided chemical investigation led to the isolation and purification of six alkaloids and three non-targeted compounds from 50 L fermentation of this endophytic fungus and their structures were elucidated as chae...

  2. The fungal endophyte Chaetomium globosum negatively affects both above- and belowground herbivores in cotton.

    Science.gov (United States)

    Zhou, Wenqing; Starr, James L; Krumm, Janice L; Sword, Gregory A

    2016-10-01

    Mutualistic plant-endophyte symbioses can benefit plants by increasing host fitness through reductions in herbivory. The fungus, Chaetomium globosum strain TAMU 520, was previously isolated as an endophyte from cotton (Gossypium hirsutum) and can be re-inoculated to systemically colonize cotton plants via seed treatment. We evaluated the potential impacts of the endophyte in cotton on plant parasitic nematodes belowground, along with piercing-sucking and chewing insects aboveground. Endophytic C. globosum inhibited root-knot nematode (Meloidogyne incognita) infection and reduced female reproduction belowground. To confirm the endophytic effect of C. globosum on root-knot nematode, a contact fungicide was applied to remove soil-borne and epiphytic C. globosum Consistent inhibition of nematode activity was observed post-fungicide treatment, with positive C. globosum colonization confirmed within plant tissues. Aboveground, endophytic C. globosum also negatively affected the fecundity of both cotton aphids (Aphis gossypii) and beet armyworms (Spodoptera exigua). Faster development rates and smaller head capsule of beet armyworm larvae were observed when fed Chaetomium-colonized plants. However, no larval weight difference was found between Chaetomium-colonized and control plants. No consistent effect on plant performance was found across experiments. Our findings illustrate how a single facultative fungal endophyte can increase plant systemic resistance against a range of invertebrate herbivores in a major crop. PMID:27451418

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

    International Nuclear Information System (INIS)

    The bifunctional enzyme catalase-phenol oxidase from S. thermophilum was crystallized by the hanging-drop vapour-diffusion method in space group P21 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 P21 and contained one tetramer per asymmetric unit

  4. INDUSTRIAL APPLICATIONS AND FUTURE PROSPECTS OF MICROBIAL XYLANASES: A REVIEW

    Directory of Open Access Journals (Sweden)

    Saurabh Sudha Dhiman

    2008-11-01

    Full Text Available Microbial enzymes such as xylanases enable new technologies for industrial processes. Xylanases (xylanolytic enzyme hydrolyze complex polysaccharides like xylan. Research during the past few decades has been dedicated to enhanced production, purification, and characterization of microbial xylanase. But for commercial applications detailed knowledge of regulatory mechanisms governing enzyme production and functioning should be required. Since application of xylanase in the commercial sector is widening, an understanding of its nature and properties for efficient and effective usage becomes crucial. Study of synergistic action of multiple forms and mechanism of action of xylanase makes it possible to use it for bio-bleaching of kraft pulp and for desizing and bio-scouring of fabrics. Results revealed that enzymatic treatment leads to the enhancement in various physical properties of the fabric and paper. This review will be helpful in determining the factors affecting xylanase production and its potential industrial applications in textile, paper, pulp, and other industries.

  5. Growth and Mycotoxin Production by Chaetomium globosum Is Favored in a Neutral pH

    Directory of Open Access Journals (Sweden)

    Matthew R. Fogle

    2008-11-01

    Full Text Available Chaetomium globosum is frequently isolated in water-damaged buildings and produces two mycotoxins called chaetoglobosins A and C when cultured on building material. In this study, the influence of ambient pH on the growth of C. globosum was examined on an artificial medium. This fungus was capable of growth on potato dextrose agar ranging in pH from 4.3 to 9.4 with optimal growth and chaetoglobosin C production occurring at a neutral pH. In addition, our results show that sporulation is favored in an acidic environment.

  6. Novel DNA barcodes for detection, idenfication and tracking of stachybotrys and chaetomium species

    DEFF Research Database (Denmark)

    Lewinska, Anna Malgorzata; Nielsen, Jakob Birkedal; Peuhkuri, Ruut Hannele;

    2014-01-01

    Detection and identification of indoor fungi in water-damaged buildings is crucial for preventi and control of fungal growth. This study focuses on a molecular method called DNA barcoding. evaluates commonly used sequences in DNA barcoding for fungal species identification Chaetomium and...... Stachybotrys. The existing DNA barcodes: ITS, SSU, LSU, B-TUB, CMD, RP and TEF-1α do not give satisfying species resolution to be considered as DNA barcodes for the two genera. Therefore, novel barcodes for them are needed. Barcode potentials, such as HOG1 a NAHA, were identified using bioinformatics and are...

  7. Epigenetic stimulation of polyketide production in Chaetomium cancroideum by an NAD(+)-dependent HDAC inhibitor.

    Science.gov (United States)

    Asai, Teigo; Morita, Shuntaro; Taniguchi, Tohru; Monde, Kenji; Oshima, Yoshiteru

    2016-01-14

    Exposure of the fungus Chaetomium cancroideum to an NAD(+)-dependent HDAC inhibitor, nicotinamide, enhanced the production of aromatic and branched aliphatic polyketides, which allowed us to isolate new secondary metabolites, chaetophenol G and cancrolides A and B. Their structures were determined using spectroscopic analyses, and their absolute configuration was elucidated by electronic circular dichroism (ECD), vibrational circular dichroism (VCD), and chemical transformations. Biosynthesis of the branched aliphatic polyketide skeletons in cancrolides A and B was evidenced by conducting a feeding experiment using compounds labeled with a (13)C stable isotope. PMID:26549741

  8. Relatedness of Thermomyces lanuginosus strains producing a thermostable xylanase.

    Science.gov (United States)

    Singh, S; Reddy, P; Haarhoff, J; Biely, P; Janse, B; Pillay, B; Pillay, D; Prior, B A

    2000-08-25

    Properties of an endo-beta-xylanase produced by a locally isolated Thermomyces lanuginosus strain SSBP was compared to seven other T. lanuginosus strains isolated from different geographical regions. Strain SSBP produced the highest xylanase activity of 59600 nkat ml(-1) when cultivated on corn cobs (maize) medium, whereas the seven other strains produced xylanase activities ranging from 6000 to 32000 nkat ml(-1). No cellulase activity was produced by the strains. Despite the variability in the production of xylanase, little difference in the other characteristics of the strains could be found. The optimal temperature and pH for xylanase production by the strains was either 40 or 50 degrees C and between pH 6 and 7, respectively. Optimal xylanase activity of the strains was observed at 70 degrees C and at pH 6 or 6.5. Culture supernatant analysis by SDS-PAGE and isoelectric focusing PAGE of all strains revealed the presence of a single 24.7 kDa and pI 3.9 xylanase. Phylogenetic analysis by PCR amplification and sequencing of the internal transcribed spacer of nuclear rRNA repeat units and 5.8S rDNA revealed no strain diversity. However, random amplified polymorphic DNA analysis pointed to greater diversity and with one primer (5'-GCCCGACGCG-3'), a relationship was established between xylanase levels and the RAPD pattern. PMID:10989171

  9. 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...

  10. Functional characterization of Penicillium occitanis Pol6 and Penicillium funiculosum GH11 xylanases

    OpenAIRE

    Berrin, Jean-Guy; Juge, Nathalie; Bhiri, Fatma; Ghorbel, Raoudha; Ellouz Chaabouni, Semia

    2013-01-01

    Xylanases are hemicellulolytic enzymes, which are responsible for the degradation of heteroxylans constituting the lignocellulosic plant cell wall. Xylanases from the GH11 family are considered as true xylanases because of their high substrate specificity. In order to study in depth a crucial difference in the thumb region between two closely related xylanases from Penicillium in terms of kinetic parameters and inhibition sensitivity, the GH11 xylanases from Penicillium occitanis Pol6 (PoXyn3...

  11. Bioconversion of (+)-valencene in submerged cultures of the ascomycete Chaetomium globosum.

    Science.gov (United States)

    Kaspera, Rüdiger; Krings, Ulrich; Nanzad, Tsevegsuren; Berger, Ralf G

    2005-06-01

    Submerged cultures of the ascomycete Chaetomium globosum oxidised the exogenous sesquiterpene (+)-valencene to nootkatone via the stereoselective generation of alpha-nootkatol. Inhibition experiments suggested that the first introduction of oxygen, the rate-limiting step of the bioconversion, may have been catalysed by a cytochrome-P450-monooxygenase. However, nootkatone was not the final metabolite: further flavour-active and inactive, non-volatile oxidation products were identified. (+)-Valencene and the flavour-active mono-oxyfunctionalised transformation products, alpha-nootkatol, nootkatone, and valencene-11,12-epoxide accumulated preferably inside the fungal cells. Di- and poly-oxygenated products, such as nootkatone-11,12-epoxide, were found solely in the culture medium, indicating an active transport of these metabolites into the extracellular compartment during (+)-valencene detoxification. These metabolic properties may have contributed to the high tolerance of the fungus towards the exogenous hydrocarbon. PMID:15602686

  12. Antifungal Metabolites Produced by Chaetomium globosum No.04, an Endophytic Fungus Isolated from Ginkgo biloba.

    Science.gov (United States)

    Zhang, Guizhen; Zhang, Yanhua; Qin, Jianchun; Qu, Xiaoyan; Liu, Jinliang; Li, Xiang; Pan, Hongyu

    2013-06-01

    The fungal endophyte Chaetomium globosum No.04 was isolated from the medicinal plant Ginkgo biloba. The crude extract of the fungus fermentation were active in the agar-diffusion tests against the phytopathogenic fungi Rhizopus stolonifer and Coniothyrium diplodiella. Further bioassay-guided chemical investigation led to the isolation and purification of six alkaloids and three non-targeted compounds from 50 L fermentation of this endophytic fungus and their structures were elucidated as chaetoglobosin A, C, D, E, G, R (1-6), ergosterol, allantoin and uracil, by means of spectroscopic analysis. Compounds 1-6 showed significant growth inhibitory activity against R. stolonifer and C. diplodiella at a concentration of 20 μg/disc. We present here, for the first time, the potent antifungal activity of chaetoglobosins from endophytic fungi against two important phytopathogenic fungi R. stolonifer and C. diplodiella. PMID:24426105

  13. Bioactive Metabolites from Chaetomium aureum: Structure Elucidation and Inhibition of the Hsp90 Machine Chaperoning Activity

    Science.gov (United States)

    Kabbaj, Fatima Zahra; Lu, Su; Faouzi, My El Abbés; Meddah, Bouchra; Proksch, Peter; Cherrah, Yahya; Altenbach, Hans-Josef; Aly, Amal H.; Chadli, Ahmed; Debbab, Abdessamad

    2014-01-01

    Chemical investigation of the EtOAc extract of the fungus Chaetomium aureum, an endophyte of the Moroccan medicinal plant Thymelaea lythroides, afforded one new resorcinol derivative named chaetorcinol, together with five known metabolites. The structures of the isolated compounds were determined on the basis of one- and two-dimensional NMR spectroscopy and high-resolution mass spectrometry as well as by comparison with the literature. All compounds were tested for their activity towards the Hsp90 chaperoning machine in vitro using the progesterone receptor (PR) and rabbit reticulocyte lysate (RRL). Among the isolated compounds, only sclerotiorin efficiently inhibited the Hsp90 machine chaperoning activity. However, sclerotiorin showed no cytotoxic effect on breast cancer Hs578T, MDA-MB-231 and prostate cancer LNCaP cell lines. Interestingly, deacetylation of sclerotiorin increased its cytotoxicity toward the tested cell lines over a period of 48h. PMID:25482429

  14. Xylanase production by Trichoderma strains in solid substrate fermentation

    Institute of Scientific and Technical Information of China (English)

    Krisztina Kovacs; George Szakacs; Lew Christopher

    2004-01-01

    @@ The importance of microbial enzymes in pulp and paper manufacturing has grown significantly in the last two decades. Solid substrate fermentation (SSF) holds tremendous potential for the production of microbial enzymes of commercial interest. SSF can be of special interest in those processes where the crude fermented product (whole SSF culture, in situ enzyme) may be used directly as the enzyme source. Xylanase preparations practically free of cellulase activity are especially useful for biobleaching of crude cellulose pulps. Thirty-nine Trichoderma isolates have been screened in SSF for xylanase production on hardwood oxygen-delignified soda-aq pulp as carbon source and enzyme inducer.Xylanase activities varied between 0 and 2200 IU/g dry matter (DM) of initial substrate. In most instances, the simultaneously produced cellulase levels were below 1.0 Filter Paper Unit (FPU) /g DM. The xylanase to cellulase activity ratio varied in the range of 5 to 3500. The three most promising isolates (TUB F-1647, TUB F-1658 and TUB F-1684) yielded xylanase activity of 2040,1300 and 1500 IU/g DM xylanase, respectively, and 0.64, 0.43 and 0.43 FPU/g DM cellulase with a xylanase to cellulase activity ratio of 3200, 3000 and 3500, respectively. Wild strains F-1647, F-1658 and F-1684 were isolated from tree bark of Maldives, soils of Peru (last two), respectively.Medium optimization experiments to enhance the xylanase yield and to increase the xylanase to cellulase ratio have also been performed.

  15. 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. PMID:11958335

  16. Synergistic effects of Bifidobacterium thermophilum RBL67 and selected prebiotics on inhibition of Salmonella colonization in the swine proximal colon PolyFermS model

    OpenAIRE

    Tanner, Sabine Amani; Chassard, Christophe; Zihler Berner, Annina; Lacroix, Christophe

    2014-01-01

    Background Probiotics and prebiotics are promising strategies to counteract Salmonella prevalence in swine. In the present study, we investigated the effects of prebiotics (fructo- (FOS), galacto- (GOS) and mannan- (MOS) oligosaccharides) and the bacteriocinogenic Bifidobacterium thermophilum RBL67 (RBL67) on Salmonella enterica subsp. enterica serovar Typhimurium N-15 (N-15) colonization using the PolyFermS in vitro continuous fermentation model simulating the swine proximal colon. Material ...

  17. Symbiodinium thermophilum sp. nov., a thermotolerant symbiotic alga prevalent in corals of the world's hottest sea, the Persian/Arabian Gulf.

    Science.gov (United States)

    Hume, B C C; D'Angelo, C; Smith, E G; Stevens, J R; Burt, J; Wiedenmann, J

    2015-01-01

    Coral reefs are in rapid decline on a global scale due to human activities and a changing climate. Shallow water reefs depend on the obligatory symbiosis between the habitat forming coral host and its algal symbiont from the genus Symbiodinium (zooxanthellae). This association is highly sensitive to thermal perturbations and temperatures as little as 1°C above the average summer maxima can cause the breakdown of this symbiosis, termed coral bleaching. Predicting the capacity of corals to survive the expected increase in seawater temperatures depends strongly on our understanding of the thermal tolerance of the symbiotic algae. Here we use molecular phylogenetic analysis of four genetic markers to describe Symbiodinium thermophilum, sp. nov. from the Persian/Arabian Gulf, a thermally tolerant coral symbiont. Phylogenetic inference using the non-coding region of the chloroplast psbA gene resolves S. thermophilum as a monophyletic lineage with large genetic distances from any other ITS2 C3 type found outside the Gulf. Through the characterisation of Symbiodinium associations of 6 species (5 genera) of Gulf corals, we demonstrate that S. thermophilum is the prevalent symbiont all year round in the world's hottest sea, the southern Persian/Arabian Gulf. PMID:25720577

  18. Chaetochromones A and B, two new polyketides from the fungus Chaetomium indicum (CBS.860.68).

    Science.gov (United States)

    Lu, Keyang; Zhang, Yisheng; Li, Li; Wang, Xuewei; Ding, Gang

    2013-01-01

    Chaetochromones A (1) and B (2), two novel polyketides, were isolated from the crude extract of fungus Chaetomium indicum (CBS.860.68) together with three known analogues PI-3(3), PI-4 (4) and SB236050 (5). The structures of these compounds were determined by HRESI-MS and NMR experiments. Chaetochromones A (1) and B (2) are a member of the polyketides family, which might originate from a similar biogenetic pathway as the known compounds PI-3 (3), PI-4 (4) and SB236050 (5). The biological activities of these secondary metabolites were evaluated against eight plant pathogens, including Alternaria alternata, Ilyonectria radicicola, Trichoderma viride pers, Aspergillus niger, Fusarium verticillioide, Irpex lacteus (Fr.), Poria placenta (Fr.) Cooke and Coriolus versicolor (L.) Quél. Compound 1 displayed moderate inhibitory rate (>60%) against the brown rot fungus Poria placenta (Fr.) Cooke, which causes significant wood decay. In addition, the cytotoxic activities against three cancer cell lines A549, MDA-MB-231, PANC-1 were also tested, without any inhibitory activities being detected. PMID:24013408

  19. Bio-control trials of Chaetomium spirale ND35 against apple canker

    Institute of Scientific and Technical Information of China (English)

    XINYa-fen; SHANGJin-jie

    2005-01-01

    A new endophytic antagonistic fungus, Chaetomium spirale ND35 from Populus tomentosa, was reported. The bio-control trials of C. spirale ND35 against the Valsa Canker of apple were preliminarily investigated. The results of dual culture on PDA plate showed that C. spirale ND35 was capable of strong antagonism against Valsa ceratosperma, and for inhibiting the mycelial growth of V. ceratosperma,.the crude extract of liquid culture of corn steep powder broth was more effective than that one of malt extract broth (MEB). The results of bio-control in greenhouse and field indicated that the disease incidence of apple tree treated with C. spirale ND35 was lower significantly than that treated by other methods. The re-isolation experiment suggested that C. spirale ND35 could colonize in stems and branches of apple trees successfully, and the ND35 colonization rate of the treatment with solid wheat bran culture was higher than that of corn steep powder broth, but the field experiment result the control effect of liquid culture of C. spirale ND35 was better than that of solid culture.

  20. Flavipin in Chaetomium globosum CDW7, an endophytic fungus from Ginkgo biloba, contributes to antioxidant activity.

    Science.gov (United States)

    Ye, Yonghao; Xiao, Yu; Ma, Liang; Li, Hongxia; Xie, Zhenglu; Wang, Minghua; Ma, Haitian; Tang, Huaiwu; Liu, Junyan

    2013-08-01

    1,2-Benzenedicarboxaldehyde-3,4,5-trihydroxy-6-methyl (flavipin) was found to be antagonistic against nematodes and fungi. Here we demonstrated that flavipin is a potent antioxidant in vitro and in vivo, which has great potential in the therapy for free radical-associated diseases. Therefore, flavipin-producing bio-source was screened from 80 endophytes in Ginkgo biloba. Seven endophytic fungi were able to synthesize antioxidant substances and identified by ITS rDNA sequences. Among them, Chaetomium globosum CDW7 was a remarkable producer of flavipin. The fermentation parameters of CDW7 were then optimized for high flavipin production. Cultured under the optimal condition (25 °C, 100/250 mL flask, 12 discs/flask, 150 rpm, pH 6.5) for 14 days, CDW7 was able to synthesize flavipin at a production of 315.5 mg/L. In addition, flavipin output was positively correlated to antioxidant activities of crude extracts with a correlation coefficient of 0.8235, indicating that flavipin was the major antioxidant component of CDW7's metabolites. These data demonstrated that CDW7 was a highly yielded bio-source of antioxidant flavipin. PMID:23740314

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

    OpenAIRE

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

    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...

  2. 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

    Directory of Open Access Journals (Sweden)

    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

  3. GH11 xylanases: Structure/function/properties relationships and applications.

    Science.gov (United States)

    Paës, Gabriel; Berrin, Jean-Guy; Beaugrand, Johnny

    2012-01-01

    For technical, environmental and economical reasons, industrial demands for process-fitted enzymes have evolved drastically in the last decade. Therefore, continuous efforts are made in order to get insights into enzyme structure/function relationships to create improved biocatalysts. Xylanases are hemicellulolytic enzymes, which are responsible for the degradation of the heteroxylans constituting the lignocellulosic plant cell wall. Due to their variety, xylanases have been classified in glycoside hydrolase families GH5, GH8, GH10, GH11, GH30 and GH43 in the CAZy database. In this review, we focus on GH11 family, which is one of the best characterized GH families with bacterial and fungal members considered as true xylanases compared to the other families because of their high substrate specificity. Based on an exhaustive analysis of the sequences and 3D structures available so far, in relation with biochemical properties, we assess biochemical aspects of GH11 xylanases: structure, catalytic machinery, focus on their "thumb" loop of major importance in catalytic efficiency and substrate selectivity, inhibition, stability to pH and temperature. GH11 xylanases have for a long time been used as biotechnological tools in various industrial applications and represent in addition promising candidates for future other uses. PMID:22067746

  4. An evolutionary route to xylanase process fitness

    Science.gov (United States)

    Palackal, Nisha; Brennan, Yali; Callen, Walter N.; Dupree, Paul; Frey, Gerhard; Goubet, Florence; Hazlewood, Geoffrey P.; Healey, Shaun; Kang, Young E.; Kretz, Keith A.; Lee, Edd; Tan, Xuqiu; Tomlinson, Geoffery L.; Verruto, John; Wong, Vicky W.K.; Mathur, Eric J.; Short, Jay M.; Robertson, Dan E.; Steer, Brian A.

    2004-01-01

    Directed evolution technologies were used to selectively improve the stability of an enzyme without compromising its catalytic activity. In particular, this article describes the tandem use of two evolution strategies to evolve a xylanase, rendering it tolerant to temperatures in excess of 90°C. A library of all possible 19 amino acid substitutions at each residue position was generated and screened for activity after a temperature challenge. Nine single amino acid residue changes were identified that enhanced thermostability. All 512 possible combinatorial variants of the nine mutations were then generated and screened for improved thermal tolerance under stringent conditions. The screen yielded eleven variants with substantially improved thermal tolerance. Denaturation temperature transition midpoints were increased from 61°C to as high as 96°C. The use of two evolution strategies in combination enabled the rapid discovery of the enzyme variant with the highest degree of fitness (greater thermal tolerance and activity relative to the wild-type parent). PMID:14718652

  5. Characterization and Purification a Specific Xylanase Showing Arabinofuranosidase Activity from Streptomyces spp. 234P-16

    OpenAIRE

    ALINA AKHDIYA; FAHRRUROZI; TRIO HENDARWIN; ANJA MERYANDINI; DEDEN SAPRUDIN; YULIN LESTARI

    2009-01-01

    Streptomyces spp 234P-16 producing xylanase was isolated from soil sample from Padang, West Sumatra, Indonesia. Crude enzyme (produced by centrifuging the culture at 14000 rpm for about 5 minutes) and purified xylanase have an optimum condition at pH 5 and 90oC. Crude xylanase have half life time of 4 hours, whereas purified xylanase have half life time of 2 ½ hours at 90oC. The molecular mass of purified xylanase was determined to be 42.4 kDa. The Arabinofuranosidase have a Km and Vmax value...

  6. Xynalase Gene Overexpression of Chaetomium cupreum in Pichia pastoris and Immobilization of Enzyme%角毛壳茵木聚糖酶基因xyn24在毕赤酵母中的高效表达及产酶的固定化研究

    Institute of Scientific and Technical Information of China (English)

    王艳君; 杨谦

    2012-01-01

    Based on the gene library of the Chaetomium cupreum mycelium, gene fragments encoding xylanase were obtained after screening, total length of 690bp encoding matureβ-1 ,4-xylanase was got through splicing and RT- PCR amplification. The recombinant methods were used to build the expression vector of xylanase in Pichia pastoris. Recombinant protein was highly expressed under the control of the AOX1 gene promoter and secreted into the medium with molecular weight of about 19.4 ku. The activity was 2.72U/mL in the measurement with chitosan as the sub- strate. SDS and metal ions Mn2+ activated the enzyme. Activity of transformants was stable after 10 generations. Xyla- nase immobilization of the preparation of chitosan micro-spherical and the nature of the free enzyme were compared. The results showed that the optimum pH of the immobilized enzyme shifted to the acidic direction, and the optimum temperature of immobilized enzyme, pH stability and thermal stability were improved.%以生防真菌角毛壳菌(Chaetomium cupreum)菌丝体时期的基因文库为基础,筛选得到编码木聚糖酶基因的片段,经拼接及RT—PCR扩增得到全长590bp的编码β—1,4-木聚糖酶基因序列,利用基因重组的方法构建可在毕赤酵母分泌表达系统中表达的木聚糖酶重组载体,并转化毕赤酵母得到重组子。在毕赤酵母醇氧化酶AOX,基因启动子的作用下,重组蛋白得到高效表达,表达蛋白分泌到培养基中,分子质量约为19.4ku;以水溶性壳聚糖为底物测得酶活为2.72U/mL。SDS及金属离子Mn2+对酶活性具有较强的激活作用;转化子经10代传代培养后酶活性稳定。通过制备的壳聚糖微球对木聚糖酶进行了固定化研究,并与游离酶的性质进行了比较。结果表明:固定化酶的最适pH范围与游离酶相比向酸性方向偏移;固定化酶的最适反应温度、酸碱稳定性及热稳定性均有所提高。

  7. THE INFLUENCE OF XYLANASE ON THE QUALITY OF BREAD

    OpenAIRE

    CHEREJI RODICA; CRETESCU IULIANA; CAPRITA RODICA

    2013-01-01

    This paper determined the quality of the bread obtained form the control flour (M) and the quality of the bread obtained from the flour with an addition of 3 different concentrations of xylanase (P1-8100 U.FXU/ 100 kg flour, P2-16200 U.FXU/ 100 kg flour, P3-24300 U.FXU/ 100 kg flour). Xylanase was used in these concentrations to establish which one is more suitable to be added in flour to obtain superior quality characteristics of the bread: higher volume, fine texture of the core, prolonging...

  8. Scytalidium thermophilum-colonized grain, corncobs and chopped wheat straw substrates for the production of Agaricus bisporus.

    Science.gov (United States)

    Sanchez, Jose E; Royse, Daniel J

    2009-02-01

    We examined the possibility of cultivating Agaricus bisporus (Ab) on various grains and agricultural by-products, with the objective of improving yield capacity of substrate pre-colonized by Scytalidium thermophilum (St). Radial growth rate (RGR) of St at 45 degrees C ranged from no growth on sterile wheat grain to 14.9 mm/d on whole oats. The linear extension rate (LER) of Ab, grown on St-colonized substrate (4 days at 45 degrees C), ranged from a low of 2.7 mm/d on 100% corncobs to 4.7 mm/d on a 50/50 mixture of ground corncobs/millet grain. Several other substrates containing wheat straw+ground corncobs+boiled millet and pre-colonized by St (4 days at 42+/-3 degrees C), were evaluated for production of Ab. The biological efficiency (BE) of production increased linearly with the addition of millet to the formula. However, substrates with millet levels 84% often were contaminated before mushroom harvest. Maximum BE (99%) and yield (21.6 kg/m(2)) were obtained on St-colonized wheat straw+2% hydrated lime supplemented with 9% commercial supplement added both at spawning and at casing. PMID:18954978

  9. Microbial xylanases and their biomedical applications: a review

    Directory of Open Access Journals (Sweden)

    Girish K. Goswami

    2013-06-01

    Full Text Available Xylanases have a great potential, mainly known for industrial applications. They can hydrolyze the xylose (Hemicellulose of plant cell wall and can be used for bio-bleaching the kraft pulp. As it reduces the requirement of harsh chemicals in the process, it can be used further to a number of bio-products with a great aggregate value. Microbial-origin xylanases can also be used in improving the nutritional quality of animal feed (e.g. food additives to poultry, piggery or fishery and indirectly affect the humans. Additionally they can be used directly in human food in bakery, clarification of juices and in xenobiotics like tobacco processing. The great value of xylanase as a bio-bleaching agent has now a new dimension of fiber digesting agent having relevance to food, drugs and cosmetics act. This review presents some important applications of Xylanases extended up to biomedical sciences. [Int J Basic Clin Pharmacol 2013; 2(3.000: 237-246

  10. Molecular cloning and characterization of multidomain xylanase from manure library

    Science.gov (United States)

    The gene (manf-x10) encoding xylanase from an environmental genomic DNA library was cloned and expressed in Escherichia coli. The encoded enzyme was predicted to be 467 amino acids with a molecular mass of 50.3 kD. The recombinant ManF-X10 was purified by HisTrap affinity column and showed activit...

  11. Polypeptides having xylanase activity and polynucleotides encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Lopez de Leon, Alfredo; Rey, Michael

    2016-05-31

    The present invention relates to isolated polypeptides having xylanase activity and isolated 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.

  12. Isolation and identification of local Bacillus isolates for xylanase biosynthesis.

    Directory of Open Access Journals (Sweden)

    Hassan Ammoneh

    2014-04-01

    Full Text Available Bacillus species are attractive industrial organisms due to their rapid growth rates leading to a short fermentation cycle and for their capacity to secrete important enzymes and proteins such as xylanase into the extracellular medium. Considering the industrial importance of xylanase, in this current study, Bacillus spp. were isolated from different soils and were screened for their xylanase production.Bacillus isolates used in this study were obtained from a national screening program carried out during 2006-2007 in which soil samples that covered areas throughout the interior of Syria were collected. The prepared inoculum from each of Bacillus isolates was aliquoted onto xylan agar plates, incubated at 30°C for 72 h and screened for xylanase synthesis.Xylanolytic isolates were selected depending on the clear zones of xylan hydrolysis. Fifteen isolates having the highest clearing zone were determined and grown in a solid state fermentation. Of the 15 isolates, three bacilli namely SY30A, SY185C and SY190E that showed maximum xylanase production, were identified using the 16S rDNA sequencing method. According to 16S rDNA gene sequence data, the closest phylogenetic neighbor for SY30A was Bacillus pumilus and for SY185C and SY190E isolates was Bacillus subtilis. Optimal pH and temperature for xylanase activity was 7.0 and 55ºC for SY30A and 6.0 and 60ºC for SY185C and SY190E, respectively. Under these conditions, the following activities were found to be around 1157 ± 58, 916 ± 46 and 794 ± 39 (U/g for SY30A, SY185C and SY190E, respectivly.Selected local Bacillus isolates were found to be a potential source of xylanase which was proven to be quite suitable for multiple biotechnological applications. These isolates might after extensive optimization steps be an alternative to commercially available strains.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    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. PMID:25346411

  15. Characterization of two truncated forms of xylanase recombinantly expressed by Lactobacillus reuteri with an introduced rumen fungal xylanase gene.

    Science.gov (United States)

    Cheng, Hsueh-Ling; Hu, Chun-Yi; Lin, Shiou-Hua; Wang, Jing-Ya; Liu, Je-Ruei; Chen, Yo-Chia

    2014-10-01

    The xylanase R8 gene (xynR8) from uncultured rumen fungi was cloned and successfully expressed in Lactobacillus reuteri. A xylanase activity of 132.1 U/mL was found in the broth of L. reuteri R8, the transformant containing pNZ3004 vector with xynR8 gene insertion. Two distinct forms of recombinant xylanase with different hydrophobicities and molecular weights were found in the broth after purification. According to the results of Western blotting, only the T7-tag, fused in the N-terminus of XynR8, could be bound to the expressed proteins, which indicated that the C-terminus of XynR8 had been truncated. These results, combined with tryptic digestion and mass spectrometry analyses, allow us to attribute the two xylanase forms to an optional cleavage of C-terminal sequences, and XynR8A, a 13 amino acid residues truncated form, and XynR8B, a 22 amino acid residues truncated form, were the main products in the extracellular fraction of L. reuteri R8. The specific activities of XynR8A and R8B were 1028 and 395 U/mg protein. Both forms of recombinant xylanase displayed a typical endoxylanase activity when they were reacted with xylan, but XynR8A demonstrated a better specific activity, catalytic efficiency and thermostability than XynR8B according to the results of enzyme characterization. These changes in enzyme properties were highly possibly caused by the present of the β-sheet in the C-terminal undeleted fragment of XynR8A. This study demonstrates that modified forms with different enzyme properties could be produced when a gene was recombinantly expressed by a L. reuteri transformant. PMID:25152410

  16. Xylanase production with xylan rich lignocellulosic wastes by a local soil isolate of Trichoderma viride

    OpenAIRE

    Goyal, Meenakshi; Kalra, K. L.; V.K. Sareen; G. Soni

    2008-01-01

    In the present study, cultural and nutritional conditions for enhanced production of xylanase by a local soil isolate of Trichoderma viride, using various lignocellulosic substrates in submerged culture fermentation have been optimized. Of the lignocellulosics used, maize straw was the best inducer followed by jowar straw for xylanase production. The highest activity achieved was between 14 to 17 days of fermentation. A continuous increase in xylanase production was observed with increasing l...

  17. OPTIMIZATION OF XYLANASE PRODUCTION FROM FREE AND IMMOBILIZED CELLS OF FUSARIUM SOLANI F7

    OpenAIRE

    Vijai Kumar Gupta; Rajeeva Gaur; Santosh Kumar Yadava; Nandan Singh Darmwal

    2009-01-01

    The aim of the present investigation was to characterize a xylanase-producing Fusarium solani isolate and to optimize cultural conditions for xylanase enzyme production from free and immobilized cells. Screening of Fusarium solani isolate was based on the diameter of the clear zone formation in oat spelt xylan agar plates. Fusarium solani isolate F7 was selected and optimized for xylanase enzyme production using cheaper substrates such as wheat straw, rice straw, rice bran, and wood husk. Max...

  18. Purification and some properties of an alkaline xylanase from alkaliphilic Bacillus sp. strain 41M-1.

    OpenAIRE

    Nakamura, S.; Wakabayashi, K; Nakai, R; Aono, R; Horikoshi, K

    1993-01-01

    An alkaliphilic Bacillus sp. strain, 41M-1, isolated from soil produced multiple xylanases extracellularly. One of these xylanases was purified to homogeneity by ammonium sulfate fractionation and anion-exchange chromatography. The moleculr mass of this enzyme (xylanase J) was 36 kDa, and the isoelectric point was pH 5.3. Xylanase J was most active at pH 9.0. The optimum temperature for the activity at pH 9.0 was around 50 degrees C. The enzyme was stable up to 55 degrees C at pH 9.0 for 30 m...

  19. THE INFLUENCE OF XYLANASE ON THE QUALITY OF BREAD

    Directory of Open Access Journals (Sweden)

    RODICA CHEREJI

    2013-12-01

    Full Text Available This paper determined the quality of the bread obtained form the control flour (M and the quality of the bread obtained from the flour with an addition of 3 different concentrations of xylanase (P1-8100 U.FXU/ 100 kg flour, P2-16200 U.FXU/ 100 kg flour, P3-24300 U.FXU/ 100 kg flour. Xylanase was used in these concentrations to establish which one is more suitable to be added in flour to obtain superior quality characteristics of the bread: higher volume, fine texture of the core, prolonging the freshness of the bread, improving the color and flavor of the bread, improving the cutting proprieties of the bread.

  20. Thermostable Xylanases of Microbial Origin: Recent Insights and Biotechnological Potential

    OpenAIRE

    S.S. Kanwar; Sunita Devi

    2012-01-01

    Xylanases are hydrolases which depolymerise the plant cell wall component-xylan, the second most abundant polysaccharide. They are mainly produced by microorganisms but can also be found in plants, marine algae, protozoans, crustaceans, insects, and snails. Because of their ability to break down xylan, these enzymes especially of microbial origin, have attracted more attention due to their potential role in pulping and bleaching processes, in food and feed industry, textile processes and orga...

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

    OpenAIRE

    Gupta Munishwar; Sharma Aparna; Dalal Sohel

    2007-01-01

    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 characterize...

  2. Fusarium graminearum produces different xylanases causing host cell death that is prevented by the xylanase inhibitors XIP-I and TAXI-III in wheat.

    Science.gov (United States)

    Tundo, Silvio; Moscetti, Ilaria; Faoro, Franco; Lafond, Mickaël; Giardina, Thierry; Favaron, Francesco; Sella, Luca; D'Ovidio, Renato

    2015-11-01

    To shed light on the role of Xylanase Inhibitors (XIs) during Fusarium graminearum infection, we first demonstrated that three out of four F. graminearum xylanases, in addition to their xylan degrading activity, have also the capacity to cause host cell death both in cell suspensions and wheat spike tissue. Subsequently, we demonstrated that TAXI-III and XIP-I prevented both the enzyme and host cell death activities of F. graminearum xylanases. In particular, we showed that the enzymatic inhibition by TAXI-III and XIP-I was competitive and only FGSG_11487 escaped inhibition. The finding that TAXI-III and XIP-I prevented cell death activity of heat inactivated xylanases and that XIP-I precluded the cell death activity of FGSG_11487 - even if XIP-I does not inhibit its enzyme activity - suggests that the catalytic and the cell death activities are separated features of these xylanases. Finally, the efficacy of TAXI-III or XIP-I to prevent host cell death caused by xylanases was confirmed in transgenic plants expressing separately these inhibitors, suggesting that the XIs could limit F. graminearum infection via direct inhibition of xylanase activity and/or by preventing host cell death. PMID:26475196

  3. A Simple Method for the Determination of Xylanase Activity on Insoluble Substrates

    Science.gov (United States)

    The propensity for a xylanase to convert insoluble (arabino)xylan into soluble oligosaccharides is an important parameter in the baking, pulp and paper, prebiotics, and biofuel industries. Current methods for determining xylanase activity on insoluble substrates are labor intensive, non-specific, or...

  4. Performance and morphometry of the intestinal mucosa of laying hens fed diets containing xylanase

    Directory of Open Access Journals (Sweden)

    KMR de Souza

    2014-09-01

    Full Text Available The objective of this study was to evaluate the effect of dietary energy level reduction and xylanase inclusion on the performance and on intestinal mucosa morphometry of two- to six-week-old laying hens. In total, 400 Hy-line W36 laying hens were distributed according to a completely randomized design in 2 x 2 factorial arrangement (energy level x inclusion of xylanase, totaling four treatments with 10 replicates of 10 birds per experimental unit. The following treatments were evaluated: positive control (balanced diet; positive control + xylanase; negative control (diet with of 100 kcal ME reduction /kg; negative control + xylanase. Body weight, weight gain, feed conversion ratio, uniformity and livability were not influenced by diets with metabolizable energy reduction and xylanase inclusion; however, the addition of xylanase to the diets resulted in shallower crypts depth and greater villus:crypt ratio in the ileum. The energy reduction of the diet associated with the supplementation of xylanase did not influence performance, but increased the feed intake of 2- to 6-week-old laying hens and increased villus height in the ileum of 6-wk-old hens. Xylanase reduces crypt depth in the ileum of 6-week-old hens.

  5. Purification and characterization of a GH11 xylanase from biobutanol-producing Clostridium beijerinckii G117.

    Science.gov (United States)

    Ng, Choong Hey; He, Jianzhong; Yang, Kun-Lin

    2015-03-01

    Most biobutanol-producing Clostridium strains are unable to ferment polysaccharides such as cellulose and xylan due to the lack of hydrolyzing enzymes. In this study, we show that Clostridium beijerinckii G117, a newly isolated biobutanol-producing strain, expresses xylanase enzyme in the presence of 1% beechwood xylan. The xylanase activity in the medium containing actively growing culture and 1% of beechwood xylan can reach up to 2.66 U/ml after 14 h of fermentation. Using salting-out and size-exclusion chromatography, we purify the crude xylanase by 8.7-fold from the supernatant with a yield of 32.2%. This purified xylanase has a molecular weight of 22.6 kDa, making it one of the smallest reported clostridial xylanases. Conserved domain analysis reveals that the xylanase belongs to glycoside hydrolase family 11 (GH11) but lacks a carbohydrate binding domain. When beechwood xylan is used as substrate for the xylanase, majority of the products are xylo-oligosaccharide (~98%), suggesting that this is an endo-1,4-β-xylanase. PMID:25564206

  6. OPTIMIZATION OF XYLANASE PRODUCTION FROM FREE AND IMMOBILIZED CELLS OF FUSARIUM SOLANI F7

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    Vijai Kumar Gupta

    2009-08-01

    Full Text Available The aim of the present investigation was to characterize a xylanase-producing Fusarium solani isolate and to optimize cultural conditions for xylanase enzyme production from free and immobilized cells. Screening of Fusarium solani isolate was based on the diameter of the clear zone formation in oat spelt xylan agar plates. Fusarium solani isolate F7 was selected and optimized for xylanase enzyme production using cheaper substrates such as wheat straw, rice straw, rice bran, and wood husk. Maximum enzyme activity was observed in wheat straw (78.32 U ml-1 for free cells and 94.68 U ml-1 for immobilized cells. Optimum pH and temperature for xylanase activity were found to be 5.5 and 30°C at 3% substrate concentration for free cells and 5.0 and 30°C at 3% substrate concentration for immobilized cells. In the purification step, 75% ammonium sulphate saturation was found to be suitable, giving maximum xylanase activity. Production of xylanase was greater from immobilized cells than from free cells. Purified xylanase from free cells yielded a single band with a molecular weight of 89kDa, while it was 92.8kDa for immobilized cells. The use of wheat straw as a major carbon source is particularly valuable, because oat spelt xylan is very expensive. The Fusarium solani F7 isolate proved to be a promising microorganism for xylanase production.

  7. Failure of the sterile air-flow component of a protected environment detected by demonstration of Chaetomium species colonization of four consecutive immunosuppressed occupants.

    Science.gov (United States)

    Woods, G L; Davis, J C; Vaughan, W P

    1988-10-01

    Four bone marrow transplant recipients consecutively occupying the same room on our Oncology-Hematology Special Care Unit (OHSCU) became colonized with Chaetomium species between January and April, 1987. These patients, aged 27 to 43 years, were immunocompromised as a result of intensive chemotherapy, and were consequently at increased risk for development of invasive fungal infection. At the time of Chaetomium colonization, all patients were febrile, two had transient new infiltrates on chest x-ray, and three were receiving amphotericin B therapy. Subsequent environmental cultures revealed Chaetomium contamination of the OHSCU air-handling system, including the HEPA (high-efficiency particulate air) filters in seven of the nine rooms comprising the unit. Because fungal colonization of HEPA filters used to create a "protective environment" for immunocompromised patients can occur and can serve as a source for patient infections, guidelines concerning proper surveillance of these HEPA filters should be established. We suggest that before a new patient enters a "protected" room, the clean side of the HEPA filter should be cultured. If fungi are recovered from that culture, we would recommend changing the filter. PMID:3066822

  8. Characterization and Purification a Specific Xylanase Showing Arabinofuranosidase Activity from Streptomyces spp. 234P-16

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    ALINA AKHDIYA

    2009-07-01

    Full Text Available Streptomyces spp 234P-16 producing xylanase was isolated from soil sample from Padang, West Sumatra, Indonesia. Crude enzyme (produced by centrifuging the culture at 14000 rpm for about 5 minutes and purified xylanase have an optimum condition at pH 5 and 90oC. Crude xylanase have half life time of 4 hours, whereas purified xylanase have half life time of 2 ½ hours at 90oC. The molecular mass of purified xylanase was determined to be 42.4 kDa. The Arabinofuranosidase have a Km and Vmax value of 1,98 mg/mL and 523 µmol/minute/mg, respectively.

  9. Isolation and Identification of the Metabolites Produced by Endophytic Fungus Chaetomium globosum ZY-22 from Ginkgo biloba%银杏内生菌Chaetomium globosum ZY-22次生代谢产物分离鉴定

    Institute of Scientific and Technical Information of China (English)

    秦建春; 白莉; 李晓明; 张雅梅; 高锦明; Hartmut laatsch

    2009-01-01

    Six metabolites cerebroside B (1),cerebroside C (2),allantoin (3),9(11)-dehyoergosterol peroxide (4) and ergosta-4,6,8,22-tetraen-3-one (5),chaetoglobosin A (6) were isolated by column chromatography from the extract of cultural mycelium of fungus Chaetomium globosum ZY-22,an endophyte in the leaves of Ginkgo biloba.Structures of them were established by spectroscopic methods.Among of them,cerebroside B,cerebroside C,allantoin were firstly obtained from endophytic fungus;The result of brine shrimp bioassay showed the mortality rates of them to Artemia salina are 1.6%,4.2%,7.4%,16.9%,12.8% and 83.6% respectively at the concentration of 10 μg/mL,chaetoglobosin A showed significant toxic effect on brine shrimp.%采用柱层析方法从银杏叶内生真菌Chaetomium globosum ZY-22的培养菌丝体提取物中分离得到脑苷脂B(1)、脑苷脂C(2)、尿囊素(3)、9(11)-去氢麦角甾醇过氧化物(4)以及4,6,8,22-四烯-3-酮-麦角甾烷(5)和球毛壳甲素(6)共6个次生代谢物;经波谱分析确定了6个化合物的结构,其中脑苷脂B、脑苷脂C和尿囊素是首次从内生真菌中得到;海虾致死试验结果显示,化合物1~6在10 μg/mL浓度下对丰年虾的致死率分别为1.6%、4.2%、7.4%、16.9%、12.8%、83.6%、表明球毛壳甲素对海虾表现出很强的毒性作用.

  10. Kinetics of Xylanase Fermentation by Recombinant Escherichia coli DH5α in Shake Flask Culture

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    Farliahati Mohd Rusli

    2009-01-01

    Full Text Available Problem statement: Interest in xylanase enzyme application has markedly increased in pulp and paper processing industries. The switch to xylanase-producing recombinant Escherichia coli DH5α pTP510 is seen here as an economic alternative towards higher productivity and easier downstream purification. Modeling of E. coli DH5α growth and enzyme secretion is thus desired for future optimization in fermentation process. Approach: Kinetics of intracellular xylanase fermentation by a recombinant E. coli DH5α was studied in shake flask culture. The effect of different medium formulations (complex, minimal and defined, initial pH (6.5, 7.0, 7.4 and 8.0 and agitation speeds (150, 200 and 250 rpm on cell growth and xylanase production were evaluated. Mathematical models based on Logistic and Luedeking-Piret equations had been proposed to describe the microbial growth and xylanase production. Results: Highest xylanase production was obtained in defined medium. Based on medium formulation, the highest cell concentration (4.59 g L-1 and xylanase production (2, 122.5 U mL-1 was obtained when (NH42HPO4 was used as the main nitrogen source, with an adjustment of the initial pH to 7.4 and agitation speed of 200 rpm. The maximum specific growth rate (µmax, growth associated xylanase production coefficient (α and non-growth associated xylanase production coefficient (β was 0.41 h-1, 474.26 U mg cell-1 and 0 U mg cell-1 h-1, respectively. Conclusion: Xylanase production was growth associated process and the enzyme secretion was greatly dependent on cell concentration and the specific growth rate of E. coli DH5α.

  11. Partial purification and characterization of xylanase produced by Penicillium expansum

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    André Luiz de Souza Querido

    2006-05-01

    Full Text Available An extracellular xylanase was found to be the major protein in the filtrate culture of Penicillium expansum when grown on 0.3 % wheat bran, which showed no xylanase multiplicity. The enzime was partial purified by.ammonium sulfate fractioning, molecular exclusion chromatography, ultrafiltration and anion exchange chromatography. The protein eluation profile showed only one form of xylanase that was partially characterized. The activity of purified xylanase was optimal at pH 5.5 and 40 ºC. The enzyme was stable at pH between 5.5 and 6.5 and temperatures between 20-40 ºC. The enzyme showed a Km of 3.03 mM and Vmax of 0.027 mumol min-1 mug -1 of protein. The enzymatic activity was increased 31 % by Mg2+ and 28 % by Al3+.Uma xilanase extracelular foi encontrada como a principal proteína na cultura filtrada de Penicillium expansum quando cultivado em farelo de trigo 0,3 %, a qual não mostrou multiplicidade. A enzima foi parcialmente purificada por fracionamento com sulfato de amônia, cromatografia de exclusão molecular, ultrafiltração e cromatografia de troca aniônica. O perfil de eluição das proteínas mostrou uma única forma de xilanase, sendo esta parcialmente caracterizada. A atividade da xilanase purificada foi ótima em pH 5.5 e à temperatura de 40 ºC. A enzima foi estável em pH entre 5,5 e 6,5 e à temperatura entre 20-40ºC. A enzima apresentou Km de 3,03 mM e Vmax de 0,027 mimol min-1 mig-1 de proteína. A atividade enzimática foi aumentada 31 % por Mg+2 e 28 % por Al+3.

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

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    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.

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

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    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.

  14. Production and Partial Characterization of an Alkaline Xylanase from a Novel Fungus Cladosporium oxysporum

    Science.gov (United States)

    Guan, Guo-Qiang; Zhao, Peng-Xiang; Zhao, Jin; Wang, Mei-Juan; Huo, Shu-Hao; Cui, Feng-Jie; Jiang, Jian-Xin

    2016-01-01

    A new fungus Cladosporium oxysporum GQ-3 producing extracellular xylanase was isolated from decaying agricultural waste and identified based on the morphology and comparison of internal transcribed spacer (ITS) rDNA gene sequence. C. oxysporum produced maximum xylanase activity of 55.92 U/mL with wheat bran as a substrate and NH4Cl as a nitrogen source. Mg2+ improved C. oxysporum xylanase production. Partially purified xylanase exhibited maximum activity at 50°C and pH 8.0, respectively, and showed the stable activity after 2-h treatment in pH 7.0–8.5 or below 55°C. Mg2+ enhanced the xylanase activity by 2% while Cu2+ had the highest inhibition ratio of 57.9%. Furthermore, C. oxysporum xylanase was resistant to most of tested neutral and alkaline proteases. Our findings indicated that Cladosporium oxysporum GQ-3 was a novel xylanase producer, which could be used in the textile processes or paper/feed industries.

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

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    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.

  16. Characterization and biotechnological application of recombinant xylanases from Aspergillus nidulans.

    Science.gov (United States)

    Maitan-Alfenas, Gabriela P; Oliveira, Mariana B; Nagem, Ronaldo A P; de Vries, Ronald P; Guimarães, Valéria M

    2016-10-01

    Two xylanases from Aspergillus nidulans, XlnB and XlnC, were expressed in Pichia pastoris, purified and characterized. XlnB and XlnC achieved maximal activities at 60°C and pH 7.5 and at 50°C and pH 6.0, respectively. XlnB showed to be very thermostable by maintaining 50% of its original activity after 49h incubated at 50°C. XlnB had its highest activity against wheat arabinoxylan while XlnC had the best activity against beechwood xylan. Both enzymes were completely inhibited by SDS and HgCl2. Xylotriose at 1mg/ml also totally inibited XlnB activity. TLC analysis showed that the main product of beechwood xylan hydrolysis by XlnB and XlnC was xylotetraose. An additive effect was shown between XlnB and XlnC and the xylanases of two tested commercial cocktails. Sugarcane bagasse saccharification results showed that these two commercial enzymatic cocktails were able to release more glucose and xylose after supplementation with XlnB and XlnC. PMID:27235731

  17. Biobleaching application of cellulase poor and alkali stable xylanase from Bacillus pumilus SV-85S

    OpenAIRE

    Nagar, Sushil; Jain, R. K.; Thakur, Vasanta Vadde; Gupta, Vijay Kumar

    2012-01-01

    The potential of extracellular alkali stable and thermo tolerant xylanase produced by Bacillus pumilus SV-85S through solid state fermentation was investigated in pulp bleaching in association with conventional bleaching using chlorine and chlorine dioxide. The biobleaching of kraft pulp with xylanase was the most effective at an enzyme dose of 10 IU/g oven dried pulp, pH 9.0 and 120 min incubation at 55 °C. Under the optimized conditions, xylanase pretreatment reduced Kappa number by 1.6 poi...

  18. Purification and preliminary characterization of a xylanase from Thermomyces lanuginosus strain SS-8

    OpenAIRE

    Shrivastava, Smriti; Shukla, Pratyoosh; Mukhopadhyay, Kunal

    2011-01-01

    Thermomyces lanuginosus SS-8 was isolated from soil samples that had been collected from near self-heating plant material and its extracellular cellulase-free xylanase purified approximately 160-fold using ion exchange chromatography and continuous elution electrophoresis. This xylanase was thermoactive (optimum temperature 60 °C) at pH 6.0 and had a molecular weight of 23.79 kDa as indicated by SDS-PAGE electrophoresis. The xylanase rapidly hydrolyzed xylan directly to xylose without the pro...

  19. A Highly Thermostable Xylanase from Stenotrophomonas maltophilia: Purification and Partial Characterization

    OpenAIRE

    Abhay Raj; Sharad Kumar; Sudheer Kumar Singh

    2013-01-01

    Seven xylanolytic bacterial strains were isolated from saw-dust dump soil. The bacterial strain X6 was selected on the basis of the highest xylanase activity with no cellulase contamination. It was identified as Stenotrophomonas maltophilia by biochemical tests and 16S rRNA gene sequencing approach. Xylanase production studies by S. maltophilia on different commercial xylans and agro-industrial residues suggested that wheat bran was the best carbon source for xylanase production (26.4 ± 0.6 I...

  20. Purification and properties of xylanase A from alkali-tolerant Bacillus sp. strain BP-23.

    OpenAIRE

    A. Blanco; Vidal, T; Colom, J F; Pastor, F I

    1995-01-01

    Xylanase A from the recently isolated Bacillus sp. strain BP-23 was purified to homogeneity. The enzyme shows a molecular mass of 32 kDa and an isoelectric point of 9.3. Optimum temperature and pH for xylanase activity were 50 degrees C and 5.5 respectively. Xylanase A was completely inhibited by N-bromosuccinimide. The main products of birchwood xylan hydrolysis were xylotetraose and xylobiose. The enzyme was shown to facilitate chemical bleaching of pulp, generating savings of 38% in terms ...

  1. Extracellular xylanase production by Pleurotus species on lignocellulosic wastes under in vivo condition using novel pretreatment.

    Science.gov (United States)

    Singh, M P; Pandey, A K; Vishwakarma, S K; Srivastava, A K; Pandey, V K

    2012-01-01

    The production of extracellular xylanase by three species of Pleurotus species i.e. P. florida, P. flabellatus and P. sajor caju was studied under in vivo condition during their cultivation on pretreated lignocellulosic wastes. Neem (Azadirachta indica) oil and ashoka (Saraca indica) leaves extract were used for pretreatment of paddy straw and wheat straw. Between these two wastes, paddy straw pretreated with neem oil, supported better xylanase production than wheat straw. Initially, xylanase production was low but it increased in subsequent days and reached at peak on 25th day of cultivation of Pleurotus species. Thereafter, there was decrease in the activity of the enzyme. On 25th day of incubation P. florida produced maximum xylanase on neem oil pretreated paddy straw i.e. 10.59 Uh—1ml—1. Among the three species, P. florida showed maximum enzyme activity followed by P. flabellatus and P. sajor caju. PMID:23273208

  2. [Screening of acidic xylanase producing strain and studies on its enzyme production conditions].

    Science.gov (United States)

    Chen, H; Zhu, J; Liang, G; Yan, Z; Zhang, S

    1999-08-01

    From 150 fungal strains, the authors found 8 strains contained mainly of xylanase activity over 100 U/mL in which the No. 149 strain was the highest xylanase producer. Which tentatively identified as Aspergillas niger. The appropriate medium composition was as follows: wheat bran hemicellulose 4%; NaNO3 1%; wheat bran 1% prepared in Mandels nutritional solution without (NH4)2SO4 and urea. After cultivated in shake-flask at 28 degrees C-32 degrees C for 60 h, the activity reached the highest value of 357.2 U/mL. The optimum pH of xylanase was 4.6 and it was stable at pH3-11. The fermented broth of strain 149 contained in addition to xylanase (relative activity 100) also included amylase(1.8), mannanase(0.98), beta-xylosidase(0.94) and cellulase(0.17). PMID:12555575

  3. Integrating a xylanase treatment into an industrial-type sequence for eucalyptus kraft pulp bleaching

    OpenAIRE

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

    2012-01-01

    The influence of a treatment with two commercial xylanases on pulp and effluents obtained after the bleaching stages in the OXAZDP (O, oxygen stage; X, xylanase treatment; A, acid stage; Z, ozone stage; D, chlorine dioxide stage; P, hydrogen peroxide stage) sequence was studied. Also, the potential saving in chlorine dioxide was assessed. The enzyme treatment was performed on pulp containing some black liquor since the operating conditions were close to the conditions used in the storage towe...

  4. Cloning and characterization of a xylanase, KRICT PX1 from the strain Paenibacillus sp. HPL-001.

    Science.gov (United States)

    Hwang, In Taek; Lim, Hee Kyung; Song, Ha Young; Cho, Soo Jin; Chang, Jong-San; Park, No-Joong

    2010-01-01

    The KRICT PX1 gene (GB: FJ380951) consisting of 996bp encoding a protein of 332 amino acids (38.1kDa) from the recently isolated Paenibacillus sp. strain HPL-001 (KCTC11365BP) has been cloned and expressed in Escherichia coli. The xylanase KRICT PX1 showed high activity on birchwood xylan, and was active over a pH range of 5.0 to 11.0, with two optima at pH 5.5 and 9.5 at 50 degrees C with K(m) value of 5.35 and 3.23, respectively. The xylanase activity was not affected by most salts, such as NaCl, LiCl, KCl, NH(4)Cl, CaCl(2), MgCl(2), MnCl(2), and CsCl(2) at 1mM, but affected by CuSO(4), ZnSO(4), and FeCl(3). One mM of EDTA, 2-mercaptoethanol, and PMSF did not affect the xylanase activity. TLC analysis of the catalyzed products after reaction with birchwood xylan revealed that xylobiose was the major product with smaller amounts of xylotriose and xylose. A similarity analysis of the amino acids in KRICT PX1 resulted 72% identity with xylanase from Geobacillus stearothermophilus (GB: ZP_03040360), 70% identity with intracellular xylanase from an uncultured bacterium (GB: AAP51133), 68% identity with endo-1-4-xylanse from Paenibacillus sp. (GB: ZP_02847150). In addition, the amino acid alignment of KRICT PX1 with glycosyl hydralase (GH) family 10 xylanases revealed a high degree of homology in highly conserved regions including the catalytic sites, and this was confirmed through PROSITE scan. These results imply that KRICT PX1 is a new xylanase gene, and this alkaline xylanase belongs to GH family 10. PMID:20493247

  5. Metabolizable energy values of diets supplemented with xylanase determined with laying hens

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    Karina Márcia Ribeiro de Souza

    2012-12-01

    Full Text Available The objective of this study was to evaluate the effect of the supplementation of xylanase in diets with reduced energy level on the apparent metabolizable energy corrected for nitrogen, determined with laying hens at 14, 36, 60 and 80 weeks of age. Four digestibility trials were conducted, using 80 Hy-line W36 laying hens aged 14, 36, 60 and 80 weeks of age. Birds were distributed in a completely randomized design in 2 × 2 factorial arrangement (energy level × inclusion of xylanase, totaling four treatments with 10 replicates of two birds each. Treatments were: positive control (balanced diet for their age; positive control + xylanase; negative control (diet with reduction of 100 kcal/kg in the level of metabolizable energy; and negative control + xylanase. Xylanase, produced by microorganism Trichoderma reesei, was added to the diets at 100 g/t (16,000 BXU/kg for diets fed at 14 weeks and 75 g/t for diets of 36, 60 and 80 weeks (12,000 BXU/kg. The data obtained were subjected to analysis of variance at 5% probability. Supplementation of xylanase promoted higher values for AME (apparent metabolizable energy and AMEn (apparent metabolizable energy corrected for nitrogen determined with 80-week-old laying hens, subjected to diet with energy level according to the nutritional requirements for their age. Supplementation of xylanase increases the matabolizability coefficient of the dietary crude protein and improves the nitrogen retention of laying hens at 14 weeks. In addition, xylanase associated with adequate levels of dietary energy promotes higher values for AME and AMEn determined with laying hens at 80 weeks of age.

  6. A Highly Thermostable Xylanase from Stenotrophomonas maltophilia: Purification and Partial Characterization

    Directory of Open Access Journals (Sweden)

    Abhay Raj

    2013-01-01

    Full Text Available Seven xylanolytic bacterial strains were isolated from saw-dust dump soil. The bacterial strain X6 was selected on the basis of the highest xylanase activity with no cellulase contamination. It was identified as Stenotrophomonas maltophilia by biochemical tests and 16S rRNA gene sequencing approach. Xylanase production studies by S. maltophilia on different commercial xylans and agro-industrial residues suggested that wheat bran was the best carbon source for xylanase production (26.4 ± 0.6 IU/mL. The studies with inorganic and organic nitrogen sources suggested yeast extract as the best support for xylanase production (25 ± 0.6 IU/mL. Maximum xylanase production was observed at initial medium pH = 8.0 (23.8 ± 0.4 IU/mL with production at pH = 7.0 and pH = 9.0 being almost comparable. Xylanase produced by S. maltophilia was purified to homogeneity using ammonium sulfate precipitation, gel filtration, and ion exchange chromatography. The final purification was 5.43-fold with recovery of 19.18%. The molecular weight of the purified xylanase protein was ~142 kDa. Both crude and purified xylanase had good stability at pH = 9.0 and 80°C with activity retention greater than 90% after 30 min incubation. The enzyme stability at high temperature and alkaline pH make it potentially effective for industrial applications.

  7. Production, purification and characterization of xylanase using alkalo-thermophilic Bacillus halodurans KR-1

    OpenAIRE

    Krityanand Kumar Mahatman; Neha Garg; Ranjeeta Chauhan; Anil Kumar

    2010-01-01

    Xylanase (EC. 3.2.1.8) has been isolated from an alkalo-thermophilic bacteria, Bacillus halodurans strain KR-1 isolated from the soil near river bed at Indore. The bacteria secreted xylanase in the growth medium in the presence of xylan. The production of the enzyme was induced in the presence of glucose, mannose, lactose and maltose whereas presence of starch, cellulose and sucrose retarded in enzyme production. The presence of casein, peptone, sodium nitrate and potassium nitrate as nitroge...

  8. Crystal Structure of Talaromyces cellulolyticus (Formerly Known as Acremonium cellulolyticus) GH Family 11 Xylanase

    OpenAIRE

    Kataoka, Misumi; Akita, Fusamichi; Maeno, Yuka; Inoue, Benchaporn; Inoue, Hiroyuki; Ishikawa, Kazuhiko

    2014-01-01

    Talaromyces cellulolyticus (formerly known as Acremonium cellulolyticus) is one of the mesophilic fungi that can produce high levels of cellulose-related enzymes and are expected to be used for the degradation of polysaccharide biomass. In silico analysis of the genome sequence of T. cellulolyticus detected seven open reading frames (ORFs) showing homology to xylanases from glycoside hydrolase (GH) family 11. The gene encoding the GH11 xylanase C (TcXylC) with the highest activity was used fo...

  9. Purification and Characterization of Aeromonas caviae ME-1 Xylanase V, Which Produces Exclusively Xylobiose from Xylan

    OpenAIRE

    Kubata, Bruno Kilunga; Suzuki, Tohru; Horitsu, Hiroyuki; Kawai, Keiichi; Takamizawa, Kazuhiro

    1994-01-01

    A xylanase, which produces exclusively xylobiose from oat spelt and birch xylans, was isolated from the culture medium of Aeromonas caviae ME-1. The enzyme (xylanase V) was purified by ammonium sulfate fractionation, hydrophobic interaction, and ion-exchange and gel filtration chromatographies. The homogeneity of the final preparation was demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and agarose gel electrofocusing. The molecular mass and isoelectric point of the x...

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

    OpenAIRE

    Song Letian; Siguier Béatrice; Dumon Claire; Bozonnet Sophie; O'Donohue Michael J

    2012-01-01

    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 scre...

  11. Removal of hexenuronic acid by xylanase to reduce adsorbable organic halides formation in chlorine dioxide bleaching of bagasse pulp.

    Science.gov (United States)

    Nie, Shuangxi; Wang, Shuangfei; Qin, Chengrong; Yao, Shuangquan; Ebonka, Johnbull Friday; Song, Xueping; Li, Kecheng

    2015-11-01

    Xylanase-aided chlorine dioxide bleaching of bagasse pulp was investigated. The pulp was pretreated with xylanase and followed a chlorine dioxide bleaching stage. The ATR-FTIR and XPS were employed to determine the surface chemistry of the control pulp, xylanase treated and chlorine dioxide treated pulps. The hexenuronic acid (HexA) could obviously be reduced after xylanase pretreatment, and the adsorbable organic halides (AOX) were reduced after chlorine dioxide bleaching. Compared to the control pulp, AOX could be reduced by 21.4-26.6% with xylanase treatment. Chlorine dioxide demand could be reduced by 12.5-22% to achieve the same brightness. The ATR-FTIR and XPS results showed that lignin and hemicellulose (mainly HexA) were the main source for AOX formation. Xylanase pretreatment could remove HexA and expose more lignin, which decreased the chlorine dioxide demand and thus reduced formation of AOX. PMID:26263004

  12. Pretreatment with xylanase and its significance in hemicellulose removal from mixed hardwood kraft pulp as a process step for viscose.

    Science.gov (United States)

    Kaur, Prabhjot; Bhardwaj, Nishi K; Sharma, Jitender

    2016-07-10

    The upturn of viscose fiber market has triggered an augmented dissolving pulp usage over the last decade. Dissolving pulp is feasible to obtain from kraft pulp after two essential steps including hemicellulose removal and subsequent pulp activation. Prerequisite of conversion being hemicellulose reduction can be gently done by using xylanase treatment prior to alkali extraction. Herein, the significance of xylanase treatment and the optimum xylanase dose required in conjunction with subsequent alkali extraction was investigated. An increase in xylanase dose prior to alkali extraction had no significant effect on pentosans while the Fock reactivity and viscosity both improved at the dose of 50AXU/g. Also, alkali extraction without xylanase pretreatment resulted in decreased Fock reactivity, alpha cellulose, brightness and viscosity of paper grade pulp. A moderate dose of xylanase prior to alkali extraction can thus be used to facilitate the hemicellulose removal while simultaneously protecting the native structure of cellulose. PMID:27106156

  13. Thermostable microbial xylanases for pulp and paper industries: trends, applications and further perspectives.

    Science.gov (United States)

    Kumar, Vishal; Marín-Navarro, Julia; Shukla, Pratyoosh

    2016-02-01

    Xylanases are enzymes with biotechnological relevance in a number of fields, including food, feed, biofuel, and textile industries. Their most significant application is in the paper and pulp industry, where they are used as a biobleaching agent, showing clear economic and environmental advantages over chemical alternatives. Since this process requires high temperatures and alkali media, the identification of thermostable and alkali stable xylanases represents a major biotechnological goal in this field. Moreover, thermostability is a desirable property for many other applications of xylanases. The review makes an overview of xylanase producing microorganisms and their current implementation in paper biobleaching. Future perspectives are analyzed focusing in the efforts carried out to generate thermostable enzymes by means of modern biotechnological tools, including metagenomic analysis, enzyme molecular engineering and nanotechnology. Furthermore, structural and mutagenesis studies have revealed critical sites for stability of xylanases from glycoside hydrolase families GH10 and GH11, which constitute the main classes of these enzymes. The overall conclusions of these works are summarized here and provide relevant information about putative weak spots within xylanase structures to be targeted in future protein engineering approaches. PMID:26754672

  14. Xylanase production with xylan rich lignocellulosic wastes by a local soil isolate of Trichoderma viride.

    Science.gov (United States)

    Goyal, Meenakshi; Kalra, K L; Sareen, V K; Soni, G

    2008-07-01

    In the present study, cultural and nutritional conditions for enhanced production of xylanase by a local soil isolate of Trichoderma viride, using various lignocellulosic substrates in submerged culture fermentation have been optimized. Of the lignocellulosics used, maize straw was the best inducer followed by jowar straw for xylanase production. The highest activity achieved was between 14 to 17 days of fermentation. A continuous increase in xylanase production was observed with increasing level of lignocellulosics in the medium and highest activity was observed with maize straw at 5% level. Xylanase production with higher levels of lignocellulosics (3 to 5%) of maize, jowar and barseem was found to be higher as compared to that with commercial xylan as carbon source. Sodium nitrate was the best nitrogen source among the six sources used. Maximum xylanase production was achieved with initial medium pH of 3.5-4.0 and incubation temperature of 25ºC.The enzyme preparation was effective in bringing about saccharification of different lignocellulosics. The xylanase production could be further improved by using alkali treated straw as carbon source. PMID:24031262

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

    Directory of Open Access Journals (Sweden)

    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.

  16. Newly Isolated Penicillium ramulosum N1 Is Excellent for Producing Protease-Resistant Acidophilic Xylanase.

    Science.gov (United States)

    Lin, Chaoyang; Shen, Zhicheng; Zhu, Tingheng; Qin, Wensheng

    2015-01-01

    Penicillium ramulosum N1 was isolated from decaying wood. This strain produces extracellular xylanases and cellulases. The highest activities of xylanases (250 U/ml) and carboxymethyl cellulose (CMCase; 6.5 U/ml) were produced when 1% barley straw was added as a carbon source. The optimum temperature and pH for xylanase activity was 55 and 3.0 °C, respectively. The xylanases exhibited strong protease resistance. CMCase revealed maximum activities at pH 3.0 and in the range of 60-70 °C. Filter paper activity was optimally active at pH 5.0 and 55 °C. The zymograms produced by the SDS-PAGE resolution of the crude enzymes indicated that there are four bands of protein with xylanase activity and three bands of proteins with endoglucanase. The results revealed that P. ramulosum N1 is a promising acidophilic and protease-resistant xylanase-producing microorganism that has great potential to be used in animal feed and food industry applications. PMID:26431535

  17. 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. PMID:27560367

  18. Xylanase Production by Bacillus circulans D1 Using Maltose as Carbon Source

    Science.gov (United States)

    Bocchini, D. A.; Gomes, E.; da Silva, R.

    Bacillus circulans D1 is a good producer of extracellular thermostable xylanase. Xylanase production in different carbon sources was evaluated and the enzyme synthesis was induced by various carbon sources. It was found that d-maltose is the best inducer of the enzyme synthesis (7.05 U/mg dry biomass at 48 h), while d-glucose and d-arabinose lead to the production of basal levels of xylanase. The crude enzyme solution is free of cellulases, even when the microorganism was cultivated in a medium with d-cellobiose. When oat spelt xylan was supplemented with d-glucose, the repressive effect of this sugar on xylanase production was observed at 24 h, only when used at 5.0 g/L, leading to a reduction of 60% on the enzyme production. On the other hand, when the xylan medium was supplemented with d-xylose (3.0 or 5.0 g/L), this effect was more evident (80 and 90% of reduction on the enzyme production, respectively). Unlike that observed in the xylan medium, glucose repressed xylanase production in the maltose medium, leading to a reduction of 55% on the enzyme production at 24 h of cultivation. Xylose, at 1.0 g/L, induced xylanase production on the maltose medium. On this medium, the repressive effect of xylose, at 3.0 or 5.0 g/L, was less expressive when compared to its effect on the xylan medium.

  19. Phylogenetic Diversity and Environment-Specific Distributions of Glycosyl Hydrolase Family 10 Xylanases in Geographically Distant Soils

    OpenAIRE

    Wang, Guozeng; Meng, Kun; Luo, Huiying; Wang, Yaru; Huang, Huoqing; Shi, Pengjun; Yang, Peilong; Zhang, Zhifang; Yao, Bin

    2012-01-01

    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 o...

  20. Kinetics of Xylanase Fermentation by Recombinant Escherichia coli DH5α in Shake Flask Culture

    OpenAIRE

    Farliahati Mohd Rusli; Mohd Shamzi Mohamed; Rosfarizan Mohamad; Ni N. Tri Puspaningsih; Arbakariya A. Ariff

    2009-01-01

    Problem statement: Interest in xylanase enzyme application has markedly increased in pulp and paper processing industries. The switch to xylanase-producing recombinant Escherichia coli DH5α pTP510 is seen here as an economic alternative towards higher productivity and easier downstream purification. Modeling of E. coli DH5α growth and enzyme secretion is thus desired for future optimization in fermentation process. Approach: Kinetics of intracellular xylanase fermentation by a recom...

  1. A new acidophilic endo-β-1,4-xylanase from Penicillium oxalicum: cloning, purification, and insights into the influence of metal ions on xylanase activity.

    Science.gov (United States)

    Liao, Hanpeng; Sun, Shaowei; Wang, Pan; Bi, Wenli; Tan, Shiyong; Wei, Zhong; Mei, Xinlan; Liu, Dongyang; Raza, Waseem; Shen, Qirong; Xu, Yangchun

    2014-07-01

    A new acidophilic xylanase (XYN11A) from Penicillium oxalicum GZ-2 has been purified, identified and characterized. Synchronized fluorescence spectroscopy was used for the first time to evaluate the influence of metal ions on xylanase activity. The purified enzyme was identified by MALDI TOF/TOF mass spectrometry, and its gene (xyn11A) was identified as an open reading frame of 706 bp with a 68 bp intron. This gene encodes a mature protein of 196 residues with a predicted molecular weight of 21.3 kDa that has the 100 % identity with the putative xylanase from the P. oxalicum 114-2. The enzyme shows a structure comprising a catalytic module family 10 (GH10) and no carbohydrate-binding module family. The specific activities were 150.2, 60.2, and 72.6 U/mg for beechwood xylan, birchwood xylan, and oat spelt xylan, respectively. XYN11A exhibited optimal activity at pH 4.0 and remarkable pH stability under extremely acidic condition (pH 3). The specific activity, K m and V max values were 150.2 U/mg, 30.7 mg/mL, and 403.9 μmol/min/mg for beechwood xylan, respectively. XYN11A is a endo-β-1,4-xylanase since it release xylobiose and xylotriose as the main products by hydrolyzing xylans. The activity of XYN11A was enhanced 155 % by 1 mM Fe(2+) ions, but was inhibited strongly by Fe(3+). The reason of enhancing the xylanase activity of XYN11A with 1 mM Fe(2+) treatment may be responsible for the change of microenvironment of tryptophan residues studied by synchronous fluorescence spectrophotometry. Inhibition of the xylanase activity by Fe(3+) was first time demonstrated to associate tryptophan fluorescence quenching. PMID:24818699

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

    International Nuclear Information System (INIS)

    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)

  3. 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...

  4. A proteomics-based study of endogenous and microbial xylanases and xylanase inhibitors associated with barley grains used for liquid feed

    DEFF Research Database (Denmark)

    Sultan, Abida

    degradation and carbohydrate catabolism. To gain a better understanding of the role of these xylanase inhibitors, the barley XIP III was expressed in a secretory Pichia pastoris system. The expressed rXIP- III with a HIS6-tag at the C -terminal was purified from the culture medium using metal affinity...

  5. Enhancing xylanase production in the thermophilic fungus Myceliophthora thermophila by homologous overexpression of Mtxyr1.

    Science.gov (United States)

    Wang, Juan; Wu, Yaning; Gong, Yanfen; Yu, Shaowen; Liu, Gang

    2015-09-01

    The xylanase regulator 1 protein in Myceliophthora thermophila ATCC42464 (MtXyr1) is 60 % homologous with that of Trichoderma reesei. However, MtXyr1's regulatory role on cellulolytic and xylanolytic genes in M. thermophila is unknown. Herein, MtXyr1 was overexpressed under the control of the MtPpdc (pyruvate decarboxylase) promoter. Compared with the wild type, the extracellular xylanase activities of the transformant cultured in non-inducing and inducing media for 120 h were 25.19- and 9.04-fold higher, respectively. The Mtxyr1 mRNA level was 300-fold higher than in the wild type in corncob-containing medium. However, the filter paper activity and endoglucanase activities were unchanged in corncob-containing medium and glucose-containing medium. The different zymograms between the transformant and the wild type were analyzed and identified by mass spectrometry as three xylanases of the glycoside hydrolase (GH) family 11. Thus, overexpression of xyr1 resulted in enhanced xylanase activity in M. thermophila. Xylanase production could be improved by overexpressing Mtxyr1 in M. thermophila. PMID:26173497

  6. Isolation, Production, and Characterization of Thermotolerant Xylanase from Solvent Tolerant Bacillus vallismortis RSPP-15

    Directory of Open Access Journals (Sweden)

    Rajeeva Gaur

    2015-01-01

    Full Text Available Sixty bacterial strains isolated from the soils sample in the presence of organic solvent were screened for xylanase production. Among them, strain RSPP-15 showed the highest xylanase activity which was identified as Bacillus vallismortis. The isolate showed maximum xylanase production (3768 U/mL in the presence of birch wood xylan and beef extract at 55°C pH 7.0 within 48 h of incubation. The enzyme activity and stability were increased 181.5, 153.7, 147.2, 133.6, and 127.9% and 138.2, 119.3, 113.9, 109, and 104.5% in the presence of Co2+, Ca2+, Mg+2, Zn+2, and Fe+3 ions (10 mM. Xylanase activity and stability were strongly inhibited in the presence of Hg and Cu ions. The enzyme was also stable in the presence of 30% of n-dodecane, isooctane, n-decane, xylene, toluene, n-hexane, n-butanol, and cyclohexane, respectively. The presence of benzene, methanol, and ethanol marginally reduced the xylanase stability, respectively. This isolate may be useful in several industrial applications owing to its thermotolerant and organic solvent resistance characteristics.

  7. Crystal structure of Talaromyces cellulolyticus (formerly known as Acremonium cellulolyticus) GH family 11 xylanase.

    Science.gov (United States)

    Kataoka, Misumi; Akita, Fusamichi; Maeno, Yuka; Inoue, Benchaporn; Inoue, Hiroyuki; Ishikawa, Kazuhiko

    2014-10-01

    Talaromyces cellulolyticus (formerly known as Acremonium cellulolyticus) is one of the mesophilic fungi that can produce high levels of cellulose-related enzymes and are expected to be used for the degradation of polysaccharide biomass. In silico analysis of the genome sequence of T. cellulolyticus detected seven open reading frames (ORFs) showing homology to xylanases from glycoside hydrolase (GH) family 11. The gene encoding the GH11 xylanase C (TcXylC) with the highest activity was used for overproduction and purification of the recombinant enzyme, permitting solving of the crystal structure to a resolution of 1.98 Å. In the asymmetric unit, two kinds of the crystal structures of the xylanase were identified. The main structure of the protein showed a β-jelly roll structure. We hypothesize that one of the two structures represents the open form and the other shows the close form. The changing of the flexible region between the two structures is presumed to induce and accelerate the enzyme reaction. The specificity of xylanase toward the branched xylan is discussed in the context of this structural data and by comparison with the other published structures of xylanases. PMID:25138599

  8. Purification and preliminary characterization of a xylanase from Thermomyces lanuginosus strain SS-8.

    Science.gov (United States)

    Shrivastava, Smriti; Shukla, Pratyoosh; Mukhopadhyay, Kunal

    2011-12-01

    Thermomyces lanuginosus SS-8 was isolated from soil samples that had been collected from near self-heating plant material and its extracellular cellulase-free xylanase purified approximately 160-fold using ion exchange chromatography and continuous elution electrophoresis. This xylanase was thermoactive (optimum temperature 60 °C) at pH 6.0 and had a molecular weight of 23.79 kDa as indicated by SDS-PAGE electrophoresis. The xylanase rapidly hydrolyzed xylan directly to xylose without the production of intermediary xylo-oligosaccharides within 15 min of incubation under optimum conditions. This trait of rapidly degrading xylan to xylose as a sole end-product could have biotechnological potential in degradation of agro-wastes for bioethanol manufacturing industry. PMID:22558544

  9. Construction of a highly active xylanase displaying oleaginous yeast: comparison of anchoring systems.

    Directory of Open Access Journals (Sweden)

    Sophie Duquesne

    Full Text Available Three Yarrowia lipolytica cell wall proteins (YlPir, YlCWP1 and YlCBM were evaluated for their ability to display the xylanase TxXYN from Thermobacillus xylanilyticus on the cell surface of Y. lipolytica. The fusion proteins were produced in Y. lipolytica JMY1212, a strain engineered for mono-copy chromosomal insertion, and enabling accurate comparison of anchoring systems. The construction using YlPir enabled cell bound xylanase activity to be maximised (71.6 U/g. Although 48% of the activity was released in the supernatant, probably due to proteolysis at the fusion zone, this system is three times more efficient for the anchoring of TxXYN than the YlCWP1 system formerly developed for Y. lipolytica. As far as we know it represents the best displayed xylanase activity ever published. It could be an attractive alternative anchoring system to display enzymes in Y. lipolytica.

  10. 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.

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

    International Nuclear Information System (INIS)

    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)

  12. Production, partial purification and characterization of xylanase using Nicotiana tabacum leaf dust as substrate.

    Science.gov (United States)

    Acharya, Komal P; Shilpkar, Prateek

    2016-03-01

    Isolated Bacillus sp. was used in the present study for production of xylanase from Nicotiana tabacum leaf dust. The strain was able to give a maximum of 1.77 Uml⁻¹ xylanase activity under optimized fermentation conditions which was further increased upto 2.77 Uml⁻¹ after extraction and partial purification of enzyme. After partial purification, the enzyme was characterized and it gave the highest xylanase activity at pH 7.0, when 0.2 ml enzyme was incubated with 2.0% substrate (Nicotiana tabacum leaf dust) for 60 min at 60°C. Saccharification study of Nicotiana tabacum leaf dust with partially purified enzyme revealed that 18.4% reducing sugar was released in 20 hrs incubation, and TLC and HPTLC analysis showed that xylose and glucose sugars were obtained after hydrolysis of substrate. FTIR analysis confirmed decomposition of substrate. PMID:27097451

  13. Study on Screening and Cultivation Conditions of Xylanase-Producing Alkalophilic Bacterial

    Institute of Scientific and Technical Information of China (English)

    Han Xiao-fang; Zheng Lian-shuang; Xie Yi-min

    2004-01-01

    An xylanase producting alkalophilic Bacillus NT-9 was obtaind by the screening method of transparent zone on the selective medium, and the effects of carbon source and nitrogen source on xylanase production were studied. The medium composed of xylose 1.5%, (NH4)2SO4 0.25%, K2HPO4 0.1%, MgSO4·7H2O 0.02%, with the initial pH of 10, was suggested to be optimal for the enzyme production in this study. When cultivatied at 37 ℃ for 72 h, the enzyme activity elaborated by the strain may reach as high as 10.5 U/mL. The xylanase produced by Bacillus NT-9 was a constituent enzyme.

  14. Bioprocess and biotecnology: effect of xylanase from Aspergillus niger and Aspergillus flavus on pulp biobleaching and enzyme production using agroindustrial residues as substract

    OpenAIRE

    de Alencar Guimaraes, Nelciele Cavalieri; Sorgatto, Michele; Peixoto-Nogueira, Simone de Carvalho; Betini, Jorge Henrique Almeida; Zanoelo, Fabiana Fonseca; Marques, Maria Rita; de Moraes Polizeli, Maria de Lourdes Teixeira; Giannesi, Giovana C

    2013-01-01

    This study compares two xylanases produced by filamentous fungi such as A. niger and A. flavus using agroindustrial residues as substract and evaluated the effect of these enzymes on cellulose pulp biobleaching process. Wheat bran was the best carbon source for xylanase production by A. niger and A. flavus. The production of xylanase was 18 and 21% higher on wheat bran when we compare the xylanase production with xylan. At 50°C, the xylanase of A. niger retained over 85% activity with 2 h of ...

  15. Exploration of Potential Actinomycetes from CIFOR Forest Origin as Antimicrobial, Antifungus, and Producing Extracellular Xylanase

    Directory of Open Access Journals (Sweden)

    Sipriyadi Sipriyadi

    2016-03-01

    Full Text Available This study aimed to isolate and explore the actinomycetes of CIFOR forest origin as an antimicrobial and antifungal agent, to produce an extracellular xylanase, and to identify isolates based on 16S rRNA gene sequences. Actinomycetes were isolated using Humic-acid Vitamin-B agar (HV media. Actinomycetes colonies that grow on the medium HV was subsequently purified by growing them on yeast malt agar (YMA media, then an antagonistic test of selected bacteria against Bacillus sp., Escherichia coli, Fusarium oxysporum, and Sclerotium sp was performed. Xylanase activity test was detected by observing a clear zone, followed by identification. Total of 35 isolates of actinomycetes isolated based on their colony morphology characteristics and diverse types of spore chains showed Streptomyces spp. of isolates CFR-06, CFR-15, CFR-17, CFR-18, and CFR-19 were able to inhibit the growth of Bacillus sp.. The highest inhibition zone has a diameter of 10.1 mm (isolate CFR-17. Isolates CFR-01 and CFR-15 were able to inhibit the growth of E. coli with the highest inhibition zone diameter of 5.1 mm (isolate CFR-15. Isolates CFR-29 and CFR-12 were able to inhibit the growth of F. oxysporum while isolate CFR-35 were able to inhibit the growth of Sclerotium sp.. Xylanase activity test showed that isolates CFR-12, CFR-20, CFR-22, CFR-24, CFR-25, CFR-30, CFR-33, CFR-34 have an ability to produce extracellular xylanase enzyme. Actinomycetes isolate (Xyl_22 as a potential xylanase enzyme producer was closely related with Streptomyces drozdowicii by the maximum similarity of 99%.How to CiteSipriyadi, S., Lestari, Y., Wahyudi, A., & Meryandini, A. (2016. Exploration Potential CIFOR Forest actinomycetes origin as Antimicrobial, Anti Fungus and Producing Enzymes Extracellular Xylanase. Biosaintifika: Journal of Biology & Biology Education, 8(1, 94-102.

  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. PMID:25604952

  17. 角毛壳菌生物防治相关基因的筛选%Selection of Biocontrol-related Genes from Chaetomium cupreum

    Institute of Scientific and Technical Information of China (English)

    张海燕; 杨谦

    2007-01-01

    采用环保的生物防治技术取代传统的化学防治技术已经在植物病害防治领域中达成共识。角毛壳菌(Chaetomium cupreum)属于子囊菌亚门毛壳菌属,是一种有效的生物防治菌,对一些常见的植物病原菌的防治效果好,例如:粉红镰刀菌(Fusarium roseum)、苹果黑星菌(Venturia inequalis)、稻梨孢(Pyricularia oryzae)、灰葡萄孢(Botrytis cinerea)等。

  18. PRODUCTION OF SINGLE CELL PROTEIN, ESSENTIAL AMINO ACIDS, AND XYLANASE BY PENICILLIUM JANTHINELLUM

    Directory of Open Access Journals (Sweden)

    Mala B. Rao

    2010-11-01

    Full Text Available Microbial biomass having 46% crude protein content and enriched with essential amino acids as well as extracellular xylanase activity (100-150 IU/ml was produced by an efficient fungal strain, Penicillium janthinellum (NCIM St-F-3b. Optimization studies for maximum xylanase and biomass production showed that the fungus required a simple medium containing bagasse hemicellulose as carbon source and ammonium sulphate as the nitrogen source. Therefore bagasse, which is a waste product of the sugar industry, can be efficiently used in microbioal biomass protein preparation for animal feed.

  19. Purification and Properties of β-1, 4-Xylanase from Aeromonas caviae W-61

    OpenAIRE

    Viet, Dung Nguyen; Kamio, Yoshiyuki; Abe, Naoki; Kaneko, Jun; Izaki, Kazuo

    1991-01-01

    Aeromonas caviae W-61, which was isolated from water samples at the Faculty of Agriculture, Tohoku University, produced β-1, 4-xylanase (1,4-β-d-xylan xylanohydrolase; EC 3.2.1.8) extracellularly. The xylanase was purified to homogeneity by using DEAE-Sephadex A-50, CM-Sephadex C-50, and Sephadex G-100 column chromatographies. The molecular weight of the purified enzyme was estimated to be 22,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point of the enzyme...

  20. Purification and characterization of a thermostable xylanase from Bacillus stearothermophilus T-6.

    OpenAIRE

    Khasin, A; Alchanati, I; Shoham, Y.

    1993-01-01

    Bacillus stearothermophilus T-6 produces an extracellular xylanase that was shown to optimally bleach pulp at pH 9 and 65 degrees C. The enzyme was purified and concentrated in a single adsorption step onto a cation exchanger and is made of a single polypeptide with an apparent M(r) of 43,000 (determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis). Xylanase T-6 is an endoxylanase that completely degrades xylan to xylose and xylobiose. The pIs of the purified protein were 9 a...

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

    OpenAIRE

    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 whe...

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

    OpenAIRE

    Giardina Thierry; Ajandouz El-Hassan; Bonnin Estelle; Desseaux Véronique; Tauzin Alexandra; Lafond Mickael

    2011-01-01

    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...

  3. EFFECT OF XYLANASE PRETREATMENT OF WOOD CHIPS ON FIBER SEPARATION IN THE CTMP REFINING PROCESS

    Directory of Open Access Journals (Sweden)

    Xiaochun Lei

    2008-08-01

    Full Text Available The effect of xylanase treatment of eucalyptus wood chips on chip refining and fiber properties was investigated. The fiber separation region and fiber surface structure were observed with SEM, TEM, and AFM. The fiber length and fines were analyzed with a Bauer-McNett classifier and optical image analysis of flowing suspensions (FQA. The results showed that xylanase degraded and hydrolyzed some xylan in the fiber wall, thus loosening the fiber wall structure. Therefore, in the subsequent refining process, fiber separation occurred in the secondary wall. This resulted in fibers with less lignin and extractives on the surface, which will benefit the interfiber bonding.

  4. Purification and characterization of thermoalkalophilic xylanase isolated from the Enterobacter sp. MTCC 5112

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Bhosle, N.B.

    for industrial applications then the enzyme need to be thermophilic and alkalophilic in nature. However, most of the xylanases known to date are optimally active at temperatures below 50 ?C and are active in acidic or neutral pH [37,30,39]. Conversely, only a... medium range (14.3 to 97.4 kDa) molecular weight markers (Banglore Genei Pvt., India). Proteins were visualized by staining with Coomassie brilliant blue. 2.12. Zymogram analysis Zymogram for xylanase was carried out using SDS-PAGE electrophoresis...

  5. PENGGUNAAN XILANASE Streptomyces sp. 45 I-3 AMOBIL UNTUK HIDROLISIS XILAN TONGKOL JAGUNG [Immobilization of Extracellular Xylanase from Streptomyces sp. 45 I-3 for Hydrolysis of Corncob Xylan

    Directory of Open Access Journals (Sweden)

    Anja Meryandini1,2*

    2009-06-01

    Full Text Available Xylan extraction from corncob is done by using alkaline as solvent. Xylan extraction from corncob could give the yields as 10.9%. One percent of corncob xylan is used as substrate to produce the xylanase, compared to oatspelt xylan. Immobilization of xylanase was performed using 1% EudragitTM S100 solution (w/v, with 5:1 volume ratio of xylanase and 1 % EudragitTM S100 (w/v. Activity of the immobilized xylanase was decreased to 23.97% compared with free xylanase. Immobilized xylanase have optimum pH and temperature at 6.0 and 40C respectively, have also thermal stability at 30–40C for an hour. Immobilized xylanase could be reused, but its activity decreased to 52.38% after 3 times application.

  6. Classification, mode of action and production strategy of xylanase and its application for biofuel production from water hyacinth.

    Science.gov (United States)

    Uday, Uma Shankar Prasad; Choudhury, Payel; Bandyopadhyay, Tarun Kanti; Bhunia, Biswanath

    2016-01-01

    Xylanases are classified under glycoside hydrolase families which represent one of the largest groups of commercial enzymes. Depolymerizing xylan molecules into monomeric pentose units involves the synergistic action of mainly two key enzymes which are endo-β-xylanase and β-xylosidase. Xylanases are different with respect to their mode of action, substrate specificities, biochemical properties, 3D structure and are widely produced by a spectrum of bacteria and fungi. Currently, large scale production of xylanase can be produced through the application of genetic engineering tool which allow fast identification of novel xylanase genes and their genetic variations makes it an ideal enzymes. Due to depletion of fossil fuel, there is urgent need to find out environment friendly and sustainable energy sources. Therefore, utilisation of cheap lignocellulosic materials along with proper optimisation of process is most important for cost efficient ethanol production. Among, various types of lignocellulosic substances, water hyacinth, a noxious aquatic weed, has been found in many tropical. Therefore, the technological development for biofuel production from water hyacinth is becoming commercially worthwhile. In this review, the classification and mode of action of xylanase including genetic regulation and strategy for robust xylanase production have been critically discussed from recent reports. In addition various strategies for cost effective biofuel production from water hyacinth including chimeric proteins design has also been critically evaluated. PMID:26529189

  7. Purification and characterization of a low molecular weight xylanase from solid-state cultures of Aspergillus fumigatus Fresenius

    OpenAIRE

    Silva Claudio Henrique Cerri e; Puls Jurgen; Sousa Marcelo Valle de; Ferreira Filho Edivaldo Ximenes

    1999-01-01

    A xylan-degrading enzyme (xylanase II) was purified to apparent homogeneity from solid-state cultures of Aspergillus fumigatus Fresenius. The molecular weight of xylanase II was found to be 19 and 8.5 kDa, as estimated by SDS-PAGE and gel filtration on FPLC, respectively. The purified enzyme was most active at 55 °C and pH 5.5. It was specific to xylan. The apparent Km and Vmax values on soluble and insoluble xylans from oat spelt and birchwood showed that xylanase II was most active on solub...

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

    OpenAIRE

    Kamble, Rajashri D.; Jadhav, Anandrao R.

    2012-01-01

    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 an...

  9. Simple, Sensitive Zymogram Technique for Detection of Xylanase Activity in Polyacrylamide Gels

    OpenAIRE

    Royer, John C.; Nakas, J. P.

    1990-01-01

    A method capable of detecting as little as 0.11 U of xylanase activity in polyacrylamide gels was developed. The method entails incubation of protein gels in contact with substrate gels containing unmodified xylan, followed by immersion of substrate gels in 95% ethanol. Resulting zymograms contain transparent bands corresponding to enzymatic activity against an opaque background.

  10. Production of crude xylanase from Thermoascus aurantiacus CBMAI 756 aiming the baking process.

    Science.gov (United States)

    Oliveira, Denise S; Meherb-Dini, Carolina; Franco, Célia M L; Gomes, Eleni; Da-Silva, Roberto

    2010-09-01

    In recent years, the baking industry has focused its attention on substituting several chemical compounds with enzymes. Enzymes that hydrolyze nonstarch polysaccharides, such as xylanase, lead to the improvement of rheological properties of dough, loaf specific volume, and crumb firmness. The purpose of this study was to find a better solid-state fermentation substrate to produce high levels of xylanase and low levels of protease and amylase, which are enzymes involved in bread quality, from Thermoascus aurantiacus CBMAI 756. Wheat bran, corncob, and corn straw were used as energy sources. The enzyme extract of corncob showed high xylanase activity (130 U/mL) and low amylase and protease activity (<1 and 15 U/mL, respectively). This enzyme profile may be more profitable for the baking industry, because it results in a slower degradation of gluten. Our results confirm this finding, because the enzyme obtained by fermentation in corncob resulted in a gluten with a higher specific volume than all the other substrates that were tested. The crude xylanase presented maximum activity at a pH of 5, and the optimum temperature was 75 °C. It was stable up to 70 °C for an hour and at a pH range from 4 to 10. PMID:21535524

  11. Reduction in acute ecotoxicity of paper mill effluent by sequential application of xylanase and laccase.

    Directory of Open Access Journals (Sweden)

    Saurabh Sudha Dhiman

    Full Text Available In order to reduce the ecotoxicity of paper mill, four different enzymatic pretreatment strategies were investigated in comparison to conventional chemical based processes. In strategy I, xylanase-aided pretreatment of pulp was carried out, and in strategy II, xylanase and laccase-mediator systems were used sequentially. Moreover, to compare the efficiency of Bacillus stearothermophilus xylanase and Ceriporiopsis subvermispora laccase in the reduction of ecotoxicity and pollution, parallel strategies (III and IV were implemented using commercial enzymes. Conventional C(DE(OPD(1D(2 (C(D, Cl(2 with ClO2; EOP, H2O2 extraction; D1 and D2, ClO2 and X/XLC(DE(OPD(1D(2 (X, xylanase; L, laccase sequences were employed with non-enzymatic and enzymatic strategies, respectively. Acute toxicity was determined by the extent of inhibition of bioluminescence of Vibrio fischeri with different dilutions of the effluent. Two-fold increase was observed in EC50 values for strategy I compared to the control process. On the other hand, sequential application of commercial enzymes resulted in higher acute toxicity compared to lab enzymes. In comparison to the control process, strategy II was the most efficient and successfully reduced 60.1 and 25.8% of biological oxygen demand (BOD and color of effluents, respectively. We report for the first time the comparative analysis of the ecotoxicity of industrial effluents.

  12. Reduction in acute ecotoxicity of paper mill effluent by sequential application of xylanase and laccase.

    Science.gov (United States)

    Dhiman, Saurabh Sudha; Garg, Gaurav; Sharma, Jitender; Kalia, Vipin C; Kang, Yun Chan; Lee, Jung-Kul

    2014-01-01

    In order to reduce the ecotoxicity of paper mill, four different enzymatic pretreatment strategies were investigated in comparison to conventional chemical based processes. In strategy I, xylanase-aided pretreatment of pulp was carried out, and in strategy II, xylanase and laccase-mediator systems were used sequentially. Moreover, to compare the efficiency of Bacillus stearothermophilus xylanase and Ceriporiopsis subvermispora laccase in the reduction of ecotoxicity and pollution, parallel strategies (III and IV) were implemented using commercial enzymes. Conventional C(D)E(OP)D(1)D(2) (C(D), Cl(2) with ClO2; EOP, H2O2 extraction; D1 and D2, ClO2) and X/XLC(D)E(OP)D(1)D(2) (X, xylanase; L, laccase) sequences were employed with non-enzymatic and enzymatic strategies, respectively. Acute toxicity was determined by the extent of inhibition of bioluminescence of Vibrio fischeri with different dilutions of the effluent. Two-fold increase was observed in EC50 values for strategy I compared to the control process. On the other hand, sequential application of commercial enzymes resulted in higher acute toxicity compared to lab enzymes. In comparison to the control process, strategy II was the most efficient and successfully reduced 60.1 and 25.8% of biological oxygen demand (BOD) and color of effluents, respectively. We report for the first time the comparative analysis of the ecotoxicity of industrial effluents. PMID:25058160

  13. 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.

    % Composition of amino acid from amino acid sequence of xylanase enzyme from Bacillus subtilis Cho40 Amino acid composition Alanine (Ala (A) 15 7.1% Arginine (Arg) (R) 6 2.8% Asparagine (Asn) (N) 19 9.0% Aspartic acid (Asp) (D...

  14. 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.

    bleaching of sugarcane bagasse pulp by a 60 min treatment at 55oC, resulting in a decrease of 10 kappa numbers and a 30% reduction in consumption of chlorine during bleaching process. The culture filtrate showed peaks of xylanase activity at acidic pH (3...

  15. Production, purification and characterization of xylanase using alkalo-thermophilic Bacillus halodurans KR-1

    Directory of Open Access Journals (Sweden)

    Krityanand Kumar Mahatman

    2010-07-01

    Full Text Available Xylanase (EC. 3.2.1.8 has been isolated from an alkalo-thermophilic bacteria, Bacillus halodurans strain KR-1 isolated from the soil near river bed at Indore. The bacteria secreted xylanase in the growth medium in the presence of xylan. The production of the enzyme was induced in the presence of glucose, mannose, lactose and maltose whereas presence of starch, cellulose and sucrose retarded in enzyme production. The presence of casein, peptone, sodium nitrate and potassium nitrate as nitrogen source in the growth medium resulted in more xylanase production, whereas presence of ammonium sulfate, ammonium nitrate and yeast extract resulted in lesser enzyme production. The enzyme has been partially purified using sodium sulfate fractionation, DEAE-cellulose and Sephadex G-200 chromatographies. The molecular weight of the enzyme has been found to be 45±02 kDa as determined by Sephadex G-200 chromatography as well as sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme protein is monomeric exhibiting maximum activity at pH 9.0. The optimum temperature for exhibiting maximum activity has been found to be 40oC. The metal ions viz. Mg2+ and Fe2+ when present in the enzyme assay medium stimulated the xylanase activity, whereas Hg2+, Co2+ and Mn2+ strongly inhibited the enzyme activity. The Km value for birchwood xylan was calculated to be 12.0 g/l.

  16. Expression of xylanases of anaerobic rumen fungi depending on carbon source in medium

    Czech Academy of Sciences Publication Activity Database

    Novotná, Zuzana; Fliegerová, Kateřina; Šimůnek, Jiří

    Clermont - Ferrand: INRA, 2008. s. 1-1. [6th INRA - RRI SYMPOSIUM: Gut microbiome . 18.06.2010 - 20.06.2008, Clermont - Ferrand] Institutional research plan: CEZ:AV0Z50450515 Keywords : xylanases * fungi * carbon source Subject RIV: EH - Ecology, Behaviour

  17. 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

    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 f...

  18. 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 ...

  19. Biobleaching of pulp from oil palm empty fruit bunches with laccase and xylanase.

    Science.gov (United States)

    Martín-Sampedro, R; Rodríguez, A; Ferrer, A; García-Fuentevilla, L L; Eugenio, M E

    2012-04-01

    Laccase and xylanase were tested for their suitability for biobleaching of soda-anthraquinone pulp from oil palm empty fruit bunches (EFB). An enzymatic stage with xylanase (X) and/or laccase (L) was incorporated before the alkaline extraction stage (E) and the hydrogen peroxide bleaching stage (P). Compared with controls, the LEP sequence resulted in an improvement of optical properties (brightness and colorimetric properties) and a reduction of the kappa number. When xylanase and laccase were used jointly, no improvement was detected, however, when the xylanase application preceded the laccase stage, the beneficial effects of laccase were boosted. Thus, the final XLEP bleached pulp showed a kappa number of 5.4 and a brightness of 60.5% ISO, although the hydrogen peroxide consumption increased (77.0% vs. 64.5% and 73.8% for EP and LEP respectively). Finally, after subjecting the bleached pulps to accelerated ageing, the best optical properties were observed in the XLEP pulp. PMID:22349193

  20. The Production of Fungal Mannanase, Cellulase and Xylanase Using Palm Kernel Meal as a Substrate

    Directory of Open Access Journals (Sweden)

    Nisa SAE-LEE

    2007-01-01

    Full Text Available Extracellular enzymes including mannanase, cellulase and xylanase from Aspergillus wentii TISTR 3075, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei TISTR 3080 and Penicillium sp. were investigated. The enzymes were produced in solid-state fermentation using palm kernel meal (PKM as a substrate. All fungal strains produced mainly mannanase. A maximum activity of 24.9 U/g koji was observed in A. wentii TISTR 3075 with a specific activity of 1.5 U/mg protein. During PKM fermentation, there was also found low concomitantly of cellulase and xylanase activities with high mannanase activity in all strains. The degradation of non-starch polysaccharides (NSPs in PKM by these fungal strains was indicated by the increased mannanase, cellulase and xylanase activities which correlated with the increase in reducing sugar content and pH profiles during PKM fermentation. PKM was shown to be more suitable for production of mannanase than cellulase and xylanase for all strains because of the high content of mannan as an inducer in PKM. Increases in enzyme yield might be obtained by optimizing the culture conditions.

  1. Xylanase production by Aspergillus awamori under solid state fermentation conditions on tomato pomace

    Directory of Open Access Journals (Sweden)

    Marcelo A. Umsza-Guez

    2011-12-01

    Full Text Available In this work, tomato pomace, a waste abundantly available in the Mediterranean and other temperate climates agro-food industries, has been used as raw material for the production of some hydrolytic enzymes, including xylanase, exo-polygalacturonase (exo-PG, cellulase (CMCase and α-amylase. The principal step of the process is the solid state fermentation (SSF of this residue by Aspergillus awamori. In several laboratory experiments, maximum xylanase and exo-PG activities were measured during the first days of culture, reaching values around 100 and 80 IU/gds (international units of enzyme activity per gram of dried solid, respectively. For CMCase and α-amylase production remained almost constant along fermentation, with average values of 19 and 21.5 IU/gds, respectively. Experiments carried out in a plate-type bioreactor at lab scale showed a clear positive effect of aeration on xylanase and CMCase, while the opposite was observed for exo-PG and α-amylase. In general, xylanase was the enzyme produced in higher levels, thus the optimum conditions for the determination of the enzyme activity was characterized. The xylanase activity shows an optimum pH of 5 and an optimum temperature of 50 ºC. The enzyme is activated by Mg2+, but strongly inhibited by Hg2+ and Cu2+. The enzymatic activity remains quite high if the extract is preserved in a range of pH from 3 to 10 and a temperature between 30 ºC to 40 ºC.

  2. Screening of xylanase activity of Streptomyces albidoflavus PSM-3n isolated from Uttarakhand

    Directory of Open Access Journals (Sweden)

    Pushpendra Sharma

    2013-07-01

    Full Text Available Background: Awareness towards the environmental pollution had made the evolution of green technology by which enzymes got special attention in industries. The enzymes replaced chemical catalysts in manufacturing various chemicals, agricultural and pharmaceutical products. Material and methods: Actinomycetes were isolated and screened for their ability to produce xylanase. For the most promising isolate, selection of media, effect of pH, temperature, metal ions, and detergents on enzyme production and activity was studied. Results: Out of 29 isolates, 22 isolates showed xylanase activity. Out of 22 xylanase producing isolate, 05 isolates were selected for secondary screening on the basis of their clear zone size. The most promising isolate PSM-3n was identified as Streptomyces albidoflavus. It produces maximum enzyme (xylanase in media Horikoshi and Ikura having carbon and nitrogen sources as oat meal and urea respectively. The optimum pH and temperature for the enzyme production was 4.0 and 45°C respectively. The enzyme activity was found maximum at temperature 50°C and enhanced in the presence of Fe3+ ions. There was a reduction in the enzyme activity in the presence of detergents like SDS, tween-20 and tween-80. The enzyme was fairly stable at 50°C for 1 h. Conclusion: The enzyme produced by the isolate PSM-3n is fairly heat stable and highly acid stable. The activity of the enzyme was increased in presence of Fe3+ ions while decreased in presence of SDS. Therefore, further studies are required for purification of xylanase for its application potential in pulp bioleaching processes and in the functional food industry.

  3. 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.

  4. Monocentric and polycentric anaerobic fungi produce structrally related cellulases and xylanases

    Energy Technology Data Exchange (ETDEWEB)

    Li, Xin-Liang; Chen, Huizhong; Ljungdahl, L.G. [Univ. of Georgia, Athens, GA (United States)

    1997-02-01

    Cellulase and xylanase cDNAs were isolated from a cDNA library of the polycentric anaerobic fungus Orpinomyces sp. strain PC-2 constructed in Escherichia coli. The cellulase cDNA (celB) was 1.8 kb long with an open reading frame (ORF) coding for a polypeptide of 471 amino acids, and the xylanase cDNA (xynA) was 1.2 kb long with an ORF encoding a polypeptide of 362 amino acids. Single transcripts of 1.9 kb for celB and 1.5 kb for xynA were detected in total RNA of Orpinomyces grown on Avicel. Genomic DNA regions coding for CelA and XynA were devoid of introns. The enzymes were highly homologous (80 to 85% identity) to the corresponding enzymes of the monocentric anaerobic fungus Neocallimastix patriciarum and, like those, contained in addition to a catalytic domain, a noncatalytic repeated peptide domain (NCRPD). The Orpinomyces xylanase contained one catalytic domain and thus differed from the Neocallimastix xylanase, which had two similar catalytic domains. Two peptides corresponding to the catalytic domain and the NCRPD of XynA were synthesized, and antibodies against them were raised and affinity column purified. The antibodies against the catalytic domain peptide reacted specifically with the xylanases of Orpinomyces and Neocallimastix, while the antibodies against the NCRPD reacted with many (at least eight) extracellular proteins of Orpinomyces and Neocallimastix, suggesting that the NCRPD is present in a number of polypeptides. 36 refs., 8 figs., 2 tabs.

  5. Purification and characterization of a low molecular weight xylanase from solid-state cultures of Aspergillus fumigatus Fresenius

    Directory of Open Access Journals (Sweden)

    Silva Claudio Henrique Cerri e

    1999-01-01

    Full Text Available A xylan-degrading enzyme (xylanase II was purified to apparent homogeneity from solid-state cultures of Aspergillus fumigatus Fresenius. The molecular weight of xylanase II was found to be 19 and 8.5 kDa, as estimated by SDS-PAGE and gel filtration on FPLC, respectively. The purified enzyme was most active at 55 °C and pH 5.5. It was specific to xylan. The apparent Km and Vmax values on soluble and insoluble xylans from oat spelt and birchwood showed that xylanase II was most active on soluble birchwood xylan. Studies on hydrolysis products of various xylans and xylooligomers by xylanase II on HPLC showed that the enzyme released a range of products from xylobiose to xylohexaose, with a small amount of xylose from xylooligomers, and presented transferase activity.

  6. Heterologous Expression of Endo-1,4-beta-xylanaseA from Phanerochaete chrysosporium in Pichia pastoris

    OpenAIRE

    Huy, Nguyen Duc; Thiyagarajan, Saravanakumar; Son, Yu-Lim; Park, Seung-Moon

    2011-01-01

    The cDNA of endo-1,4-β-xylanaseA, isolated from Phaenerocheate chrysosporium was expressed in Pichia pastoris. Using either the intrinsic leader peptide of XynA or the α-factor signal peptide of Saccharomyces cerevisiae, xylanaseA is efficiently secreted into the medium at maximum concentrations of 1,946 U/L and 2,496 U/L, respectively.

  7. Biochemical properties of xylanases from a thermophilic fungus, Melanocarpus albomyces, and their action on plant cell walls

    OpenAIRE

    Prabhu, Ashok K; Maheshwari, Ramesh

    1999-01-01

    Melanocarpus albomyces, a thermophilic fungus isolated from compost by enrichment culture in a liquid medium containing sugarcane bagasse, produced cellulase-free xylanase in culture medium. The fungus was unusual in that xylanase activity was inducible not only by hemicellulosic material but also by the monomeric pentosan unit of xylan but not by glucose. Concentration of bagasse-grown culture filtrate protein followed by size-exclusion and anion-exchange chromatography separated four xylana...

  8. Purification and properties of thermostable xylanase and beta-xylosidase produced by a newly isolated Bacillus stearothermophilus strain.

    OpenAIRE

    Nanmori, T; Watanabe, T.; Shinke, R; Kohno, A; Kawamura, Y.

    1990-01-01

    We isolated a thermophilic bacterium that produces both xylanase and beta-xylosidase. Based on taxonomical research, this bacterium was identified as Bacillus stearothermophilus. Each extracellular enzyme was separated by hydrophobic chromatography by using a Toyopearl HW-65 column, followed by gel filtration with a Sephacryl S-200 column. Each enzyme in the culture was further purified to homogeneity (62-fold for xylanase and 72-fold for beta-xylosidase) by using a fast protein liquid chroma...

  9. Improvement of xylanase production by Aspergillus niger XY-1 using response surface methodology for optimizing the medium composition

    Institute of Scientific and Technical Information of China (English)

    Yao-xing XU; Yan-li LI; Shao-chun XU; Yong LIU; Xin WANG; Jiang-wu TANG

    2008-01-01

    Objective: To study the optimal medium composition for xylanase production by Aspergillus niger XY-1 in solid-state fermentation (SSF). Methods: Statistical methodology including the Plackett-Burman design (PBD) and the central composite design (CCD) was employed to investigate the individual crucial component of the medium that significantly affected the enzyme yield. Results: Firstly, NaNO3, yeast extract, urea, Na2CO3, MgSO4, peptone and (NH4)2SO4 were screened as the significant factors positively affecting the xylanase production by PBD. Secondly, by valuating the nitrogen sources effect, urea was proved to be the most effective and economic nitrogen source for xylanase production and used for further optimization.Finally, the CCD and response surface methodology (RSM) were applied to determine the optimal concentration of each sig-nificant variable, which included urea, Na2CO3 and MgSO4. Subsequently a second-order polynomial was determined by mul-tiple regression analysis. The optimum values of the critical components for maximum xylanase production were obtained as follows: x1 (urea)=0.163 (41.63 g/L), x2 (Na2CO3)=-1.68 (2.64 g/L), x3 (MGSO4)=1.338 (10.68 g/L) and the predicted xylanase value was 14374.6 U/g dry substrate. Using the optimized condition, xylanase production by Aspergillus niger XY-1 after 48 h fermentation reached 14637 U/g dry substrate with wheat bran in the shake flask. Conclusion: By using PBD and CCD, we obtained the optimal composition for xylanase production by Aspergillus niger XY-1 in SSF, and the results of no additional expensive medium and shortened fermentation time for higher xylanase production show the potential for industrial utilization.

  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. Novel xylanases from Simplicillium obclavatum MTCC 9604: comparative analysis of production, purification and characterization of enzyme from submerged and solid state fermentation

    OpenAIRE

    Roy, Saugata; Dutta, Tanmay; Sarkar, Tuhin Subhra; Ghosh, Sanjay

    2013-01-01

    The production of extracellular xylanase by a newly isolated fungus Simplicillium obclavatum MTCC 9604 was studied in solid-state and submerged fermentation. Multiple xylanases and endoglucanases were produced by the strain during growth on wheat bran in solid state fermentation (SSF). A single xylanase isoform was found to be produced by the same fungus under submerged fermentation (SF) using wheat bran as sole carbon source. Enzyme activity, stability and the protein yield were much higher ...

  12. 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.

  13. Industrial textile effluent decolourization in stirred and static batch cultures of a new fungal strain Chaetomium globosum IMA1 KJ472923.

    Science.gov (United States)

    Manai, Imène; Miladi, Baligh; El Mselmi, Abdellatif; Smaali, Issam; Ben Hassen, Aida; Hamdi, Moktar; Bouallagui, Hassib

    2016-04-01

    The treatment of an industrial textile effluent (ITE) was investigated by using a mono-culture of a novel fungal strain Chaetomium globosum IMA1. This filamentous fungus was selected based on its capacity for dye removal via the biodegradation mechanism. The respirometric analysis showed that C. globosum IMA1 was resistant to an indigo concentration up to 700 mg equivalent COD/L. The decolourization of the ITE by C. globosum was performed in static and stirred batch systems. The better lignin peroxidase (LiP), laccase and the manganese peroxidase (MnP) productions were 829.9 U/L, 83 U/L and 247.8 U/L, respectively since 3-5 days under a stirred condition. Therefore, the chemical oxygen demand (COD) and colors (OD620) removal yields reached 88.4% and 99.8%, respectively. Fourier transforms infrared spectroscopy (FTIR) analysis of the treated effluent showed that the decolourization was due to the degradation and the transformation of dye molecules. However, spectrophotometric examination showed that the complete dye removal was through fungal adsorption (8%), followed by degradation (92%). PMID:26775156

  14. 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

  15. Pseudomonas sp. xylanase for clarification of Mausambi and Orange fruit juice

    Science.gov (United States)

    Sharma, Pawan Kumar; Chand, Duni

    2012-07-01

    Xylanase can be usd for many Industrial applications and juice clarification is one of them. Pseudomonas sp. xylanase was used for fruit juice clarification in free State. Maximum amount of juice clarification was in case of Mausambi juice was observed at 40 C∞ and 52 hours, in case of free enzyme treated juice there is 46.9% increase in clarity and 1.7 fold increase in reducing sugars of the juice and enzyme dose was optimized as 8U with maximum flow rate of 6 ml/min at this dose. In case of orange juice in free enzyme treated juice maximum clarity was observed at 40 C∞ and 52 hours, juice was found to be 42.14 % clear with increase of 1.9 fold of reducing sugars, enzyme dose optimized was 8.06U with maximum flow rate of 0.86 ml/min.

  16. APPLICATIONS OF XYLANASE, LACCASE ENZYME AND HIGH POWER ULTRASOUND ON DIFFERENT NON-WOOD PLANTS

    OpenAIRE

    Panjiyar, Niraj

    2011-01-01

    The purpose of this bachelor`s thesis was to analyze the effect of properly applied microbial enzymes on non-wood plants delignification, improved fibre flexibility, fibrillation, removal of xylan and facilitated contaminant. The enzyme plays important role in digestion of fibres and removes lignin content of pulp. In the experimental part, two different non-woody plants were experimented: flax, straw. The enzymes Laccase and Xylanase were used. The amount of enzyme was tested in four p...

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

    OpenAIRE

    Georgi Todorov Dobrev; Boriana Yordanova Zhekova

    2012-01-01

    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 ac...

  18. Screening of xylanase activity of Streptomyces albidoflavus PSM-3n isolated from Uttarakhand

    OpenAIRE

    Pushpendra Sharma; Vijay Kumar; Bindu Naik; Gajraj Singh Bisht

    2013-01-01

    Background: Awareness towards the environmental pollution had made the evolution of green technology by which enzymes got special attention in industries. The enzymes replaced chemical catalysts in manufacturing various chemicals, agricultural and pharmaceutical products. Material and methods: Actinomycetes were isolated and screened for their ability to produce xylanase. For the most promising isolate, selection of media, effect of pH, temperature, metal ions, and detergents on enzyme pro...

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

    OpenAIRE

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

    2016-01-01

    Abstract 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...

  20. 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. PMID:24045195

  1. Partial purification and characterization of Xylanase from Trichoderma viride produced under SSF

    Directory of Open Access Journals (Sweden)

    M Irfan

    2012-03-01

    Full Text Available Summary: In the present study xylanase enzyme was produced from Trichoderma viride in solid state fermentation using sugarcane bagasse as a substrate. The whole fermentation process was carried out in 250ml Erlenmeyer flask at 30oC for seven days of fermentation period. The enzyme was partially purified by ammonium sulphate (60% fractionation followed by dialysis. The partially purified enzyme was further characterized showing optimum pH and temperature of 5.0 and 50oC respectively. Metal profile of the enzyme showed that it was stimulated by FeSO4 (134%, CaCl2 (129%, BaCl2 (105%, MgSO4 (113%, MnCl2 (102% or AgCl (107% and it was strongly inhibited by EDTA (26% or HgSO4 (32%. Industrial Relevance: In the present study, xylanase enzyme was produced and characterized from Trichoderma viride in solid state fermentation using cheap substrate. This enzyme is very helpful in industrial sector especially in pulp and paper industry, food industry and also in bioethanol production. Pilot scale production of this enzyme in industries can reduce the import cost of the enzyme and make the whole process cost effective. Keywords: Partial purification; Characterization; Xylanase; Trichoderma viride; SSF

  2. 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. PMID:27209456

  3. Novel alkali-thermostable xylanase from Thielaviopsis basicola (MTCC 1467): Purification and kinetic characterization.

    Science.gov (United States)

    Goluguri, Baby Rani; Thulluri, Chiranjeevi; Addepally, Uma; Shetty, Prakasham Reddy

    2016-01-01

    A novel extracellular alkali-thermostable xylanase was purified to an apparent homogeneity from the submerged fermented culture filtrate of Thielaviopsis basicola MTCC 1467, wherein, the fungus was fed with rice straw as prime carbon source. SDS-PAGE analysis of the xylanase showcased molecular weight of ∼ 32 kDa. This extracellular protein macromolecule had maximum xylanolytic activity at pH 5.5 and 60°C, and was stable in the range of pH 5.0-10.0 for 5 days retaining >70% activity. The enzyme was stable at 30-50°C for 5h retaining >85% activity and further by retaining 70% activity at 60°C for 2h. The enzyme deactivation constants (kd) were in range of 0.41-1.3. The kinetic experiments specified that the enzyme had Km and Vmax values of 1.447 ± 0.22 mg mL(-1) and 60.04 ± 1.25 IU mL(-1), respectively, for xylan. The purified xylanase was significantly inhibited by Cu(2+) and Zn(2+) (∼ 58%), whilst Ca(2+) and Na(+) ions displayed partial inhibition (<8%) Intriguingly, the K(+) and Mn(2+) ions enhanced the activity by about ∼ 10%. Both SDS and EDTA reduced its activity by ∼ 20%. PMID:26526179

  4. Low-cost carbon sources for the production of a thermostable xylanase by Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Ana Cláudia Elias Pião Benedetti

    2013-01-01

    Full Text Available A strain of the filamentous fungus Aspergillus niger was isolated and shown to possess extracellular xylanolytic activity. These enzymes have biotechnological potential and can be employed in various industries. This fungus produced its highest xylanase activity in a medium made up of 0.1% CaCO3 , 0.5% NaCl, 0.1% NH4 Cl, 0.5% corn steep liquor and 1% carbon source, at pH 8.0. A lowcost hemicellulose residue (powdered corncob proved to be an excellent inducer of the A. niger xylanolytic complex. Filtration of the crude culture medium with suspended kaolin was ideal for to clarify the extract and led to partial purification of the xylanolytic activity. The apparent molecular mass of the xylanase was about 32.3 kDa. Maximum enzyme activity occurred at pH 5.0 and 55-60ºC. Apparent Km was 10.41 ± 0.282 mg/mL and Vmax was 3.32 ± 0.053 U/mg protein, with birchwood xylan as the substrate. Activation energy was 4.55 kcal/mol and half-life of the crude enzyme at 60ºC was 30 minutes. Addition of 2% glucose to the culture medium supplemented with xylan repressed xylanase production, but in the presence of xylose the enzyme production was not affected.

  5. 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.

  6. Immobilization of xylanase purified from Bacillus pumilus VLK-1 and its application in enrichment of orange and grape juices.

    Science.gov (United States)

    Kumar, Lalit; Nagar, Sushil; Mittal, Anuradha; Garg, Neelam; Gupta, Vijay Kumar

    2014-09-01

    This study was conducted to evaluate the efficacy of purified free and immobilized xylanase in enrichment of fruit juices. Extracellular xylanase produced from Bacillus pumilus VLK-1 was purified to apparent homogeneity by 15.4-fold with 88.3 % recovery in a single step using CM-Sephadex C-50. Purified xylanase showed a single band on SDS-polyacrylamide gel with a molecular mass of 22.0 kDa. The purified enzyme was immobilized on glutaraldehyde-activated aluminum oxide pellets and the immobilization process parameters were optimized statistically through response surface methodology. The bound enzyme displayed an increase in optimum temperature from 60 to 65 ºC and pH from 8.0 to 9.0. The pH and temperature stability of the enzyme was also enhanced after immobilization. It could be reused for 10 consecutive cycles with 58 % residual enzyme activity. The potential of purified xylanase (free and immobilized) in juice enrichment from grape (Vitis amurensis) and orange (Citrus sinensis) pulps has been investigated. The optimization of this process using free xylanase revealed maximum juice yield, clarity and reducing sugar on treatment with 20 IU/g fruit pulp for 30 min at 50 ºC. Treatment of both the fruit pulps with xylanase under optimized conditions resulted in an increase in juice yield, clarity, reducing sugars, titratable acidity, and filterability but a decline in turbidity and viscosity. Immobilized enzyme was more effective in improving juice quality as compared to its soluble counterpart. The results showed B. pumilus VLK-1 xylanase, in both free and immobilized form, as a potential candidate for use in fruit juice enrichment. PMID:25190829

  7. Does release of encapsulated nutrients have an important role in the efficacy of xylanase in broilers?

    Science.gov (United States)

    Khadem, A; Lourenço, M; Delezie, E; Maertens, L; Goderis, A; Mombaerts, R; Höfte, M; Eeckhaut, V; Van Immerseel, F; Janssens, G P J

    2016-05-01

    The non-starch polysaccharides (NSPs) in cell walls can act as a barrier for digestion of intracellular nutrients. This effect is called "cage effect." Part of the success of fibrolytic enzymes in broiler feed is assumed to be attributed to cage effect reduction. Further, changes in viscosity and potential prebiotic action should also be considered. The aim of this study was to gain insight into the relative importance of the cage effect in xylanase efficacy in broilers. Using a 2×2 factorial design, 24 pens with 30 Ross 308 male chicks were fed corn-soy based diets consisting of normal and freeze-thawed (5 d at -18°C) corn, both with and without xylanase. The freeze-thaw method was used to eliminate the cage effect, whereas a corn-based diet was used to exclude viscosity effects. Body weights (BW), feed intake (FI), and feed conversion ratio (FCR) were determined at d 13, 26, and 39. A balance study was executed at the end of the growing phase. These birds were euthanized at d 34 (non-fasted) to determine the viscosity of digesta, blood metabolites, intestinal morphology, and microbiota composition. During the finisher period, there was a significant interaction between enzyme supplementation and freeze-thawing for FCR, in which FCR was improved by freeze-thawed corn and tended to be improved by normal corn+enzyme compared with the control group. The improvement in performance (finisher period) of freeze-thawed corn and xylanase coincided with increased gut absorption of glucose (based on postprandial plasma concentrations) and increased number of Clostridiumcluster IV in the caecum, and agreed with the higher gut villus height. In addition, xylanase inclusion significantly increased the postprandial plasma glycine and triglycerides concentration, and led to elevated bacterial gene copies of butyryl CoA:acetate CoA-transferase, suggesting a prebiotic effect of xylanase addition through more than just the cage effect reduction. The applied model managed to rule

  8. Production of Xylanase from Arthrobacter sp. MTCC 6915 Using Saw Dust As Substrate under Solid State Fermentation

    Directory of Open Access Journals (Sweden)

    Sevanan Murugan

    2011-01-01

    Full Text Available Saw dust was used as substrate for xylanase production from Arthrobacter sp. MTCC 6915. The study of period of incubation, temperature, pH, carbon, and nitrogen sources for xylanase production was optimized. Xylanase production was found to be optimum at an incubation period of 96 hrs (117.0 U/mL, temperature 30°C (105.0 U/mL, and pH 9.0 (102.9 U/mL. The results showed that the xylanase production was found to be higher in the presence of carboxymethylcellulose (176.4 U/mL and dextrose (126.0 U/mL. It was also observed that peptone (170.1 U/mL and beef extract (161.7 U/mL supported maximum xylanase production.The enzyme was characterized and found to be fairly active at pH 9 (764.4 U/mL and temperature 60°C (819 U/mL. Even in the present study, a major difference in the production temperature (30°C and optimal temperature (60°C of the enzyme activity was observed. However, the pH of the production media and the enzyme activity were found to be the same (pH 9.

  9. 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.

  10. The influence of some factors on β-1,4-xylanase activity of the filamentous fungus Trichoderma reesei QM9414

    Directory of Open Access Journals (Sweden)

    Alexandru Manoliu

    2012-03-01

    Full Text Available The mesophyllic fungus Trichoderma reesei (anamorph to Hypocrea jecorina is an important biotechnological tool, known for its ability to secrete large quantities of hydrolytic enzymes. Renewable biomass, such as agricultural and forest wastes are used to produce microbial enzymes in various industrial processes such as food, feed and bioethanol industries. In raw biomass materials, such as wheat straws, barley straws and maize stalks, the main polysaccharide is cellulose which is closely associated with hemicelluloses like xylan, manan and xyloguclan. In consequence, the hydrolysis of these materials requires the concerted action of several enzymes, namely cellulases and xylanases. Endo-xylanase (endo-1,4--xylanase, EC 3.2.1.8 is the key enzyme involved in xylan hydrolysis, the mainhemicellulosic component of plant cell walls. The metabolic activity and enzyme productivity of Trichoderma reesei isinfluenced by various environmental conditions. In this context, we analysed the effect of pH, cultivation period, thenature of the substrate used and the nitrogen source on enzymatic activity. The maximum xylanase yield was recorded at a initial pH of 4 (116.189 IU/ml for barley and 5 for wheat (88.578 IU/ml, respectively maize (116.583 IU/ml. The bestsubstrate for endo-xylanase activity was maize stalks (90.446 IU/ml at a a concentration of 30g/L.

  11. Bioprocess and biotecnology: effect of xylanase from Aspergillus niger and Aspergillus flavus on pulp biobleaching and enzyme production using agroindustrial residues as substract.

    Science.gov (United States)

    de Alencar Guimaraes, Nelciele Cavalieri; Sorgatto, Michele; Peixoto-Nogueira, Simone de Carvalho; Betini, Jorge Henrique Almeida; Zanoelo, Fabiana Fonseca; Marques, Maria Rita; de Moraes Polizeli, Maria de Lourdes Teixeira; Giannesi, Giovana C

    2013-01-01

    This study compares two xylanases produced by filamentous fungi such as A. niger and A. flavus using agroindustrial residues as substract and evaluated the effect of these enzymes on cellulose pulp biobleaching process. Wheat bran was the best carbon source for xylanase production by A. niger and A. flavus. The production of xylanase was 18 and 21% higher on wheat bran when we compare the xylanase production with xylan. At 50°C, the xylanase of A. niger retained over 85% activity with 2 h of incubation, and A. flavus had a half-life of more than 75 minutes. At 55°C, the xylanase produced by A. niger showed more stable than from A. flavus showing a half-life of more than 45 minutes. The xylanase activity of A. niger and A. flavus were somehow protected in the presence of glycerol 5% when compared to the control (without additives). On the biobleaching assay it was observed that the xylanase from A. flavus was more effective in comparison to A. niger. The kappa efficiency corresponded to 36.32 and 25.93, respectively. That is important to emphasize that the cellulase activity was either analyzed and significant levels were not detected, which explain why the viscosity was not significantly modified. PMID:24010038

  12. 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. PMID:24362246

  13. Optimization of endoglucanase and xylanase activities from Fusarium verticillioides for simultaneous saccharification and fermentation of sugarcane bagasse.

    Science.gov (United States)

    de Almeida, Maíra N; Guimarães, Valéria M; Falkoski, Daniel L; Paes, Guilherme B T; Ribeiro, José Ivo; Visser, Evan M; Alfenas, Rafael F; Pereira, Olinto L; de Rezende, Sebastião T

    2014-02-01

    Enzymatic hydrolysis is an important but expensive step in the production of ethanol from biomass. Thus, the production of efficient enzymatic cocktails is of great interest for this biotechnological application. The production of endoglucanase and xylanase activites from F. verticillioides were optimized in a factorial design (2(5)) followed by a CCDR design. Endoglucanase and xylanase activities increased from 2.8 to 8.0 U/mL and from 13.4 to 114 U/mL, respectively. The optimal pH and temperature were determined for endoglucanase (5.6, 80 °C), cellobiase (5.6, 60 °C), FPase (6.0, 55 °C) and xylanase (7.0, 50 °C). The optimized crude extract was applied in saccharification and fermentation of sugarcane bagasse from which 9.7 g/L of ethanol was produced at an ethanol/biomass yield of 0.19. PMID:24170331

  14. Improved Production of Aspergillus usamii endo-β-1,4-Xylanase in Pichia pastoris via Combined Strategies

    Directory of Open Access Journals (Sweden)

    Jianrong Wang

    2016-01-01

    Full Text Available A series of strategies were applied to improve expression level of recombinant endo-β-1,4-xylanase from Aspergillus usamii (A. usamii in Pichia pastoris (P. pastoris. Firstly, the endo-β-1,4-xylanase (xynB gene from A. usamii was optimized for P. pastoris and expressed in P. pastoris. The maximum xylanase activity of optimized (xynB-opt gene was 33500 U/mL after methanol induction for 144 h in 50 L bioreactor, which was 59% higher than that by wild-type (xynB gene. To further increase the expression of xynB-opt, the Vitreoscilla hemoglobin (VHb gene was transformed to the recombinant strain containing xynB-opt. The results showed that recombinant strain harboring the xynB-opt and VHb (named X33/xynB-opt-VHb displayed higher biomass, cell viability, and xylanase activity. The maximum xylanase activity of X33/xynB-opt-VHb in 50 L bioreactor was 45225 U/mL, which was 35% and 115% higher than that by optimized (xynB-opt gene and wild-type (xynB gene. Finally, the induction temperature of X33/xynB-opt-VHb was optimized in 50 L bioreactor. The maximum xylanase activity of X33/xynB-opt-VHb reached 58792 U/mL when the induction temperature was 22°C. The results presented here will greatly contribute to improving the production of recombinant proteins in P. pastoris.

  15. Cloning, Expression, and Purification of Xylanase Gene from Bacillus licheniformis for Use in Saccharification of Plant Biomass.

    Science.gov (United States)

    Zafar, Asma; Aftab, Muhammad Nauman; Din, Zia Ud; Aftab, Saima; Iqbal, Irfana; Shahid, Anam; Tahir, Arifa; Haq, Ikram Ul

    2016-01-01

    The xylanase gene (xynA) of Bacillus licheniformis 9945A was cloned and expressed in Escherichia coli BL21(DE3) using pET-22b(+) as an expression vector. The recombinant xylanase enzyme was purified by ammonium sulfate precipitation, followed by single-step immobilized metal ion affinity chromatography with a 57.58-fold purification having 138.2 U/mg specific activity and recovery of 70.08 %. Molecular weight of the purified xylanase, 23 kDa, was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was stable for up to 70 °C with a broad pH range of 4-9 pH units. The enzyme activity was increased in the presence of metal ions especially Ca(+2) and decreased in the presence of EDTA, indicating that the xylanase was a metalloenzyme. However, an addition of 1-4 % Tween 80, β-mercaptoethanol, and DTT resulted in the increase of enzyme activity by 51, 52, and 5 %, respectively. Organic solvents with a concentration of 10-40 % slightly decreased the enzyme activity. The xylanase enzyme possesses the ability of bioconversion of plant biomasses like wheat straw, rice straw, and sugarcane bagasse. Among the different tested biomasses, the highest saccharification percentage was observed with 1 % sugarcane bagasse after 72 h of incubation at 50 °C with 20 units of enzyme. The results suggest that recombinant xylanase can be used in the bioconversion of natural biomasses into simple sugars which could be further used for the production of biofuel. PMID:26438315

  16. Response surface optimization for xylanase with high volumetric productivity by indigenous alkali tolerant Aspergillus candidus under submerged cultivation

    OpenAIRE

    Garai, Debabrata; Kumar, Vineet

    2012-01-01

    In this study, a novel isolate Aspergillus candidus was employed for xylanase production using low cost agro residues. A Box-Behnken design matrix was used to optimize the influential parameters like carbon source, nitrogen source and incubation temperature for maximum xylanase production. Under optimized condition, enzyme titer level enhanced to 69 IU/ml at 48 h with volumetric productivity 1437 IU/l h. Growth and enzyme production were observed even at pH 11.0, indicating its ability to sus...

  17. ENHANCED PRODUCTION OF CELLULASE-FREE XYLANASE BY ALKALOPHILIC BACILLUS SUBTILIS ASH AND ITS APPLICATION IN BIOBLEACHING OF KRAFT PULP

    OpenAIRE

    Ashwani Sanghi; Neelam Garg; Kalika Kuhar; Kuhad, Ramesh C.; Gupta, Vijay K

    2009-01-01

    This paper reports high level production of a cellulase-free xylanase using wheat bran, a cost-effective substrate, under submerged fermentation by alkalophilic Bacillus subtilis ASH. Production of xylanase was observed even at alkaline pH up to 11.0 and temperature 60 °C, although the highest enzyme titer was recorded at neutral pH and 37 °C. The enzyme production under optimized fermentation was 1.5-fold greater than under unoptimized conditions. Pre-treatment of unbleached pulp of 10% cons...

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

    OpenAIRE

    Leandra Zafalon Jaekel; Camila Batista da Silva; Caroline Joy Steel; Yoon Kil Chang

    2012-01-01

    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 form...

  19. Characterization of a purified thermostable xylanase from Caldicoprobacter algeriensis sp. nov. strain TH7C1(T)

    OpenAIRE

    Bouanane-Darenfed, A.; Boucherba, N.; Bouacem, K.; Gagaoua, M.; Joseph, M; Kebbouche-Gana, S.; Nateche, F.; Hacene, H.; Ollivier, Bernard; Cayol, J. L.; Fardeau, Marie-Laure

    2016-01-01

    The present study investigates the purification and biochemical characterization of an extracellular thermostable xylanase (called XYN35) from Caldicoprobacter algeriensis sp. nov., strain TH7C1(T), a thermophilic, anaerobic strain isolated from the hydrothermal hot spring of Guelma (Algeria). The maximum xylanase activity recorded after 24 h of incubation at 70 degrees C and in an optimized medium containing 10 g/L mix birchwood-and oats spelt-xylan was 250 U/mL. The pure protein was obtaine...

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

    OpenAIRE

    Sanghi, Ashwani; Garg, Neelam; Gupta, V. K.; Mittal, Ashwani; R.C. Kuhad

    2010-01-01

    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 ran...

  1. Characterization and comparison of Clostridium cellulovorans endoglucanases-xylanases EngB and EngD hyperexpressed in Escherichia coli.

    OpenAIRE

    Foong, F C; Doi, R H

    1992-01-01

    By the use of a T7 expression system, endoglucanases-xylanases EngB and EngD from Clostridium cellulovorans were hyperexpressed and purified from Escherichia coli. The two enzymes demonstrated both endoglucanase and xylanase activities. The substrate specificities of both endoglucanases were similar except that EngD had four-times-greater p-nitrophenyl beta-1,4-cellobiosidase activity. The two proteins were very homologous (80%) up to the Pro-Thr-Thr region which divided the protein into -NH2...

  2. 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.

  3. Obtaining a mutant of Bacillus amyloliquefaciens xylanase A with improved catalytic activity by directed evolution.

    Science.gov (United States)

    Xu, Xin; Liu, Ming-Qi; Huo, Wen-Kang; Dai, Xian-Jun

    2016-05-01

    This study aimed to obtain xylanase exhibiting improved enzyme properties to satisfy the requirements for industrial applications. The baxA gene encoding Bacillus amyloliquefaciens xylanase A was mutated by error-prone touchdown PCR. The mutant, pCbaxA50, was screened from the mutant library by using the 96-well plate high-throughput screening method. Sequence alignment revealed the identical mutation point S138T in xylanase (reBaxA50) produced by the pCbaxA50. The specific activity of the purified reBaxA50 was 9.38U/mg, which was 3.5 times higher than that of its parent expressed in Escherichia coli BL21 (DE3), named reBaxA. The optimum temperature of reBaxA and reBaxA50 were 55°C and 50°C, respectively. The optimum pH of reBaxA and reBaxA50 were pH 6 and pH 5, respectively. Moreover, reBaxA50 was more stable than reBaxA under thermal and extreme pH treatment. The half-life at 60°C and apparent melting temperature of reBaxA50 were 9.74min and 89.15°C, respectively. High-performance liquid chromatography showed that reBaxA50 released xylooligosaccharides from oat spelt, birchwood, and beechwood xylans, with xylotriose as the major product; beechwood xylan was also the most thoroughly hydrolyzed. This study demonstrated that the S138T mutation possibly improved the catalytic activity and thermostability of reBaxA50. PMID:26992794

  4. 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

  5. Low-cost carbon sources for the production of a thermostable xylanase by Aspergillus niger

    OpenAIRE

    Ana Cláudia Elias Pião Benedetti; Eliana Dantas da Costa; Caio Casale Aragon; Andréa Francisco dos Santos; Antônio José Goulart; Derlene Attili-Angelis; Rubens Monti

    2013-01-01

    A strain of the filamentous fungus Aspergillus niger was isolated and shown to possess extracellular xylanolytic activity. These enzymes have biotechnological potential and can be employed in various industries. This fungus produced its highest xylanase activity in a medium made up of 0.1% CaCO3 , 0.5% NaCl, 0.1% NH4 Cl, 0.5% corn steep liquor and 1% carbon source, at pH 8.0. A lowcost hemicellulose residue (powdered corncob) proved to be an excellent inducer of the A. ni...

  6. Partial purification and characterization of Xylanase from Trichoderma viride produced under SSF

    OpenAIRE

    Irfan, M.; Q. Syed

    2012-01-01

    Summary: In the present study xylanase enzyme was produced from Trichoderma viride in solid state fermentation using sugarcane bagasse as a substrate. The whole fermentation process was carried out in 250ml Erlenmeyer flask at 30oC for seven days of fermentation period. The enzyme was partially purified by ammonium sulphate (60%) fractionation followed by dialysis. The partially purified enzyme was further characterized showing optimum pH and temperature of 5.0 and 50oC respectively. Metal pr...

  7. Effect of additives on adsorption and desorption behavior of xylanase on acid-insoluble lignin from corn stover and wheat straw.

    Science.gov (United States)

    Li, Yanfei; Ge, Xiaoyan; Sun, Zongping; Zhang, Junhua

    2015-06-01

    The competitive adsorption between cellulases and additives on lignin in the hydrolysis of lignocelluloses has been confirmed, whereas the effect of additives on the interaction between xylanase and lignin is not clear. In this work, the effects of additives, poly(ethylene glycol) 2000, poly(ethylene glycol) 6000, Tween 20, and Tween 80, on the xylanase adsorption/desorption onto/from acid-insoluble lignin from corn stover (CS-lignin) and wheat straw (WS-lignin) were investigated. The results indicated that the additives could adsorb onto isolated lignin and reduce the xylanase adsorption onto lignin. Compared to CS-lignin, more additives could adsorb onto WS-lignin, making less xylanase adsorbed onto WS-lignin. In addition, the additives could enhance desorption of xylanase from lignin, which might be due to the competitive adsorption between xylanase and additives on lignin. The released xylanase from lignin still exhibited hydrolytic capacity in the hydrolysis of isolated xylan and xylan in corn stover. PMID:25818260

  8. 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.

  9. Contribution of ethanol-tolerant xylanase G2 from Aspergillus oryzae on Japanese sake brewing.

    Science.gov (United States)

    Sato, Yuichiro; Fukuda, Hisashi; Zhou, Yan; Mikami, Shigeaki

    2010-12-01

    We purified three xylanase isozymes (XynF1, XynF3 and XynG2) from a solid-state Aspergillus oryzae RIB128 culture using chromatography. The results of our sake-brewing experiment, in which we used exogenously supplemented enzymes, revealed that only XynG2 improved the alcohol yield and the material utilization. The alcohol yield of the XynG2 batch displayed an increase of 4.4% in comparison to the control, and the amount of sake cake decreased by 4.6%. The contribution of XynG2 was further confirmed through our brewing experiment in which we used the yeast heterogeneously expressing fungal xylanase isozymes. Interestingly XynG1, an enzyme with a XynG2-like sequence that is more vulnerable to ethanol, did not improve the sake-mash fermentation. The stability of XynG2 in ethanol was prominent, and it retained most of its original activity after we exposed it to 80% ethanol for 30min, whereas the stability of the other isozymes in ethanol, including XynG1, was much lower (20-25% ethanol). We concluded, therefore, that the improvement of material utilization achieved with XynG2 is primarily attributable to its characteristically high stability in ethanol, thereby, effectively degrading rice endosperm cell walls under high-alcohol conditions such as a sake-mash environment. PMID:20727822

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

    International Nuclear Information System (INIS)

    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)

  11. Endo-xylanase and endo-cellulase-assisted extraction of pectin from apple pomace.

    Science.gov (United States)

    Wikiera, Agnieszka; Mika, Magdalena; Starzyńska-Janiszewska, Anna; Stodolak, Bożena

    2016-05-20

    Pectins were extracted from apple pomace with monoactive preparation of endo-xylanase and endo-cellulase. The process was conducted for 10h in conditions of pH 5.0 at 40°C, with constant shaking. Endo-xylanase application resulted in the highest extraction efficiency of pectins (19.8%). The obtained polymer was characterised by a very high molecular mass, high level of neutral sugars - mainly arabinose, galactose and glucose, and very high DM (73.4). It also contained the highest level of protein and phenols. Pectin extracted with endo-cellulase had 1.5 fold lower molecular mass but contained significantly more GalA (70.5%) of a high degree of methylation (66.3%). The simultaneous application of both enzymatic preparations resulted in their cooperation, leading to a decrease of both the extraction efficiency and the molecular mass of pectin. However, this pectin was distinguished by the highest GalA (74.7%) and rhamnose contents. PMID:26917391

  12. 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. PMID:26776003

  13. Purification and characterization of the xylanase produced by Jonesia denitrificans BN-13.

    Science.gov (United States)

    Boucherba, Nawel; Gagaoua, Mohammed; Copinet, Estelle; Bettache, Azeddine; Duchiron, Francis; Benallaoua, Said

    2014-03-01

    Jonesia denitrificans BN-13 produces six xylanases: Xyl1, Xyl2, Xyl3, Xyl4, Xyl5, and Xyl6; the Xyl4 was purified and characterized after two consecutive purification steps using ultrafiltration and anion exchange chromatography. The xylanase-specific activity was found to be 77 unit (U)/mg. The molecular weight of the Xyl4 estimated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed a monomeric isoenzyme of about 42 kDa. It showed an optimum pH value of 7.0 and a temperature of 50 °C. It was stable at 50 °C for 9.34 h. The enzyme showed to be activated by Mn(+2), β-mercaptoethanol, and dithiothreitol (DTT) with a high affinity towards birchwood xylan (with a K(m) of 1 mg ml(-1)) and hydrolysis of oat-spelt xylan with a K(m) of 1.85 mg ml(-1). The ability of binding to cellulose and/or xylan was also investigated. PMID:24425300

  14. Improvement of thermostability and activity of Trichoderma reesei endo-xylanase Xyn III on insoluble substrates.

    Science.gov (United States)

    Matsuzawa, Tomohiko; Kaneko, Satoshi; Yaoi, Katsuro

    2016-09-01

    Trichoderma reesei Xyn III, an endo-β-1,4-xylanase belonging to glycoside hydrolase family 10 (GH10), is vital for the saccharification of xylans in plant biomass. However, its enzymatic thermostability and hydrolytic activity on insoluble substrates are low. To overcome these difficulties, the thermostability of Xyn III was improved using random mutagenesis and directed evolution, and its hydrolytic activity on insoluble substrates was improved by creating a chimeric protein. In the screening of thermostable Xyn III mutants from a random mutagenesis library, we identified two amino acid residues, Gln286 and Asn340, which are important for the thermostability of Xyn III. The Xyn III Gln286Ala/Asn340Tyr mutant showed xylanase activity even after heat treatment at 60 °C for 30 min or 50 °C for 96 h, indicating a dramatic enhancement in thermostability. In addition, we found that the addition of a xylan-binding domain (XBD) to the C-terminal of Xyn III improved its hydrolytic activity on insoluble xylan. PMID:27138202

  15. 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; Nunan, Kylie J.; Madrid, Susan M.; Brinch-Pedersen, Henrik; Holm, Preben Bach; Scheller, Henrik Vibe

    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. Transg...

  16. ENHANCED PRODUCTION OF CELLULASE-FREE XYLANASE BY ALKALOPHILIC BACILLUS SUBTILIS ASH AND ITS APPLICATION IN BIOBLEACHING OF KRAFT PULP

    Directory of Open Access Journals (Sweden)

    Ashwani Sanghi

    2009-08-01

    Full Text Available This paper reports high level production of a cellulase-free xylanase using wheat bran, a cost-effective substrate, under submerged fermentation by alkalophilic Bacillus subtilis ASH. Production of xylanase was observed even at alkaline pH up to 11.0 and temperature 60 °C, although the highest enzyme titer was recorded at neutral pH and 37 °C. The enzyme production under optimized fermentation was 1.5-fold greater than under unoptimized conditions. Pre-treatment of unbleached pulp of 10% consistency with crude xylanase (6 IU/g o.d. pulp at 60 ºC for 2 h increased the final brightness by 4.9%. The enzyme treatment reduced the chlorine consumption by 28.6% with the same brightness as in the control. A reduction in kappa number and increase in viscosity was observed after enzyme pre-treatment. Scanning electron microscopy revealed loosening and swelling of pulp fibers. The strength properties viz. grammage, fiber thickness, beating degree, tensile index, breaking length, tear index and double fold of the treated pulp were improved as compared to the control pulp. This study reveals the potential of B. subtilis ASH xylanase as a biobleaching agent for the paper and pulp industry.

  17. 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...

  18. A Comparison of Polysaccharide Substrates and Reducing Sugar Methods for the Measurement of endo-1,4-β-Xylanase.

    Science.gov (United States)

    McCleary, Barry V; McGeough, Paraic

    2015-11-01

    The most commonly used method for the measurement of the level of endo-xylanase in commercial enzyme preparations is the 3,5-dinitrosalicylic acid (DNS) reducing sugar method with birchwood xylan as substrate. It is well known that with the DNS method, much higher enzyme activity values are obtained than with the Nelson-Somogyi (NS) reducing sugar method. In this paper, we have compared the DNS and NS reducing sugar assays using a range of xylan-type substrates and accurately compared the molar response factors for xylose and a range of xylo-oligosaccharides. Purified beechwood xylan or wheat arabinoxylan is shown to be a suitable replacement for birchwood xylan which is no longer commercially available, and it is clearly demonstrated that the DNS method grossly overestimates endo-xylanase activity. Unlike the DNS assay, the NS assay gave the equivalent colour response with equimolar amounts of xylose, xylobiose, xylotriose and xylotetraose demonstrating that it accurately measures the quantity of glycosidic bonds cleaved by the endo-xylanase. The authors strongly recommend cessation of the use of the DNS assay for measurement of endo-xylanase due to the fact that the values obtained are grossly overestimated due to secondary reactions in colour development. PMID:26289020

  19. 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.

  20. 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 was a...

  1. Cloning and in-silico analysis of beta-1,3-xylanase from psychrophilic yeast, Glaciozyma antarctica PI12

    Science.gov (United States)

    Nor, Nooraisyah Mohamad; Bakar, Farah Diba Abu; Mahadi, Nor Muhammad; Murad, Abdul Munir Abdul

    2015-09-01

    A beta-1,3-xylanase (EC 3.2.1.32) gene from psychrophilic yeast, Glaciozyma antarctica has been identified via genome data mining. The enzyme was grouped into GH26 family based on Carbohydrate Active Enzyme (CaZY) database. The molecular weight of this protein was predicted to be 42 kDa and is expected to be soluble for expression. The presence of signal peptide suggested that this enzyme may be released extracellularly into the marine environment of the host's habitat. This supports the theory that such enzymatic activity is required for degradation of nutrients of polysaccharide origins into simpler carbohydrates outside the environment before it could be taken up inside the cell. The sequence for this protein showed very little conservation (< 30%) with other beta-1,3-xylanases from available databases. Based on the phylogenetic analysis, this protein also showed distant relationship to other xylanases from eukaryotic origin. The protein may have undergone major substitution in its gene sequence order to adapt to the cold climate. This is the first report of beta-1,3-xylanase gene isolated from a psychrophilic yeast.

  2. Production and purification of an Endo-1,4-b-Xylanase from Humicola grisea var. thermoidea by electroelution

    OpenAIRE

    Monti Rubens; Cardello Leonardo; Custódio Marcos F.; Goulart Antonio J.; Sayama Adriana H.; Contiero Jonas

    2003-01-01

    Humicola grisea var. thermoidea produces two forms of extracellular xylanase. The component form 1 was purified using the electroelution method, due to the very small production of this extracellular enzyme. The apparent molecular mass was 61.8 kDa by SDS-PAGE.

  3. 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.

  4. Characterization of a newly identified rice chitinase-like protein (OsCLP homologous to xylanase inhibitor

    Directory of Open Access Journals (Sweden)

    Wu Jingni

    2013-01-01

    Full Text Available Abstract Background During rice blast fungal attack, plant xylanase inhibitor proteins (XIPs that inhibit fungal xylanase activity are believed to act as a defensive barrier against fungal pathogens. To understand the role of XIPs in rice, a xylanase inhibitor was cloned from rice. The expression of this gene was examined at the transcriptional/translational levels during compatible and incompatible interactions, and the biochemical activity of this protein was also examined. Results Sequence alignment revealed that the deduced amino acid sequence of OsCLP shares a high degree of similarity with that of other plant TAXI-type XIPs. However, recombinant OsCLP did not display inhibitory activity against endo-1,4-β-xylanase enzymes from Aureobasidium pullulans (A. pullulans or Trichoderma viride (T. viride. Instead, an in-gel activity assay revealed strong chitinase activity. The transcription and translation of OsCLP were highly induced when rice was exposed to pathogens in an incompatible interaction. In addition, exogenous treatment with OsCLP affected the growth of the basidiomycete fungus Rhizoctonia solani through degradation of the hyphal cell wall. These data suggest that OsCLP, which has chitinase activity, may play an important role in plant defenses against pathogens. Conclusions Taken together, our results demonstrate that OsCLP may have antifungal activity. This protein may directly inhibit pathogen growth by degrading fungal cell wall components through chitinase activity.

  5. Agar-agar entrapment increases the stability of endo-β-1,4-xylanase for repeated biodegradation of xylan.

    Science.gov (United States)

    Bibi, Zainab; Shahid, Faiza; Ul Qader, Shah Ali; Aman, Afsheen

    2015-04-01

    Microbial xylanases, specially endo-β-1,4-xylanase catalyzes the hydrolysis of xylan, is considered one of the most significant hydrolases. It has numerous applications but most extensively is utilized in paper and pulp industry as a bio-bleaching agent. Immobilization technique is comprehensively studied with the expectation of modifying and improving enzyme stability and characteristics for commercial purposes. Currently, matrix entrapment technique is applied to immobilize endo-β-1,4-xylanase within agar-agar gel beads produced by Geobacillus stearothermophilus KIBGE-IB29. Maximal enzyme immobilization yield was achieved at 2.5% of agar-agar concentration. Optimized conditions demonstrated an increase in the optimal reaction time from 05 min to 30 min and incubation temperature from 50 °C to 60 °C with reference to free enzyme whereas; no effect was observed for optimum pH. Entrapment technique uniquely changed the kinetic parameters of immobilized endo-β-1,4-xylanase (Km: 0.5074 mg min(-1) to 0.5230 mg min(-1) and Vmax: 4773 U min(-1) to 968 U min(-1)) as compared to free enzyme. However, immobilized enzyme displayed broad thermal stability and retained 79.0% of its initial activity at 80 °C up to 30 min whereas; free enzyme completely lost its activity at this temperature. With respect to economic feasibility, the immobilized enzyme showed impressive recycling efficiency up to six reaction cycles. PMID:25603143

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

    OpenAIRE

    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.

  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

    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.

  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

    International Nuclear Information System (INIS)

    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 oC, and exclusively xylobiose at 90 oC 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. Utilization of Agro-industrial Wastes for the Simultaneous Production of Amylase and Xylanase by Thermophilic Actinomycetes

    Directory of Open Access Journals (Sweden)

    Renu Singh

    2012-12-01

    Full Text Available Agro-industrial wastes such as sugarcane bagasse, wheat bran, rice bran, corn cob and wheat straw are cheapest and abundantly available natural carbon sources. The present study was aimed to production of amylase and xylanase simultaneously using agro-industrial waste as the sole carbon source. Seven thermophilic strains of actinomycete were isolated from the mushroom compost. Among of these, strain designated MSC702 having high potential to utilize agro-industrial wastes for the production of amylase and xylanase. Strain MSC702 was identified as novel species of Streptomyces through morphological characterization and 16S rRNA gene sequence. Enzyme production was determined using 1% (w/v of various agro-industrial waste in production medium containing (g/100mL: K2HPO4(0.1, (NH42SO4(0.1, NaCl (0.1, MgSO4(0.1 at pH 7.0 after incubation of 48 h at 50°C. The amylase activity (373.89 IU/mL and xylanase activity (30.15 IU/mL was maximum in rice bran. The decreasing order of amylase and xylanase activity in different type of agro-industrial wastes were found rice bran (RB > corn cob (CC > wheat bran (WB > wheat straw (WS > sugarcane bagasse (SB and rice bran (RB > wheat bran (WB > wheat straw (WS > sugarcane bagasse (SB > corn cob (CC, respectively. Mixed effect of different agro-industrial wastes was examined in different ratios. Enzyme yield of amylase and xylanase was ~1.3 and ~2.0 fold higher with RB: WB in 1:2 ratio.

  10. Ileal amino acid digestibility and performance of growing pigs fed wheat-based diets supplemented with xylanase.

    Science.gov (United States)

    Barrera, M; Cervantes, M; Sauer, W C; Araiza, A B; Torrentera, N; Cervantes, M

    2004-07-01

    Two experiments were conducted to determine the effect of supplementation of xylanase to a wheat-based diet on the apparent ileal digestibility (AID) of AA and the performance of growing pigs fed diets limiting in AA. In Exp. 1, eight pigs (average initial BW = 20.5+/-1.2 kg) fitted with a simple T-cannula at the distal ileum, were fed four diets according to a repeated 4 x 4 Latin square design. Diet 1 was a basal diet that contained 97.6% wheat. Diets 2, 3, and 4 were the basal diet supplemented with xylanase at rates of 5,500, 11,000, and 16,500 units of xylanase activity (XU), respectively (as-fed basis). There were linear and quadratic effects (0.062 lysine, 0.12% threonine, and 0.05% methionine. Diet 6 (positive control diet) was a wheat-soybean meal diet that contained 18.2% CP (as-fed basis). The total contents of lysine, threonine, and methionine were similar for Diets 5 and 6. There was a linear effect of xylanase supplementation on ADG (P = 0.093) and feed:gain ratio (P = 0.089), and a quadratic effect on ADG (P = 0.067) and feed:gain ratio (P = 0.074). But, the greatest response was obtained with the supplementation of 11,000 XU. The supplementation of lysine, threonine, and methionine to Diet 1 increased (P = 0.001) ADG and ADFI and improved (P = 0.01) feed:gain ratio. There was no difference (P = 0.508) in the performance of pigs fed the AA-supplemented or control diet. In conclusion, the supplementation of xylanase to a diet in which wheat provided the sole source of protein and energy improved the AID of AA, ADG, and feed:gain ratio; however, this improvement was very small compared with that obtained with the supplementation of synthetic amino acids. PMID:15309946

  11. 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.

  12. Expression of endo-1, 4-beta-xylanase from Trichoderma reesei in Pichia pastoris and functional characterization of the produced enzyme

    Directory of Open Access Journals (Sweden)

    He Jun

    2009-06-01

    Full Text Available Abstract Background In recent years, xylanases have attracted considerable research interest because of their potential in various industrial applications. The yeast Pichia pastoris can neither utilize nor degrade xylan, but it possesses many attributes that render it an attractive host for the expression and production of industrial enzymes. Results The Xyn2 gene, which encodes the main Trichoderma reesei Rut C-30 endo-β-1, 4-xylanase was cloned into the pPICZαA vector and expressed in Pichia pastoris. The selected P. pastoris strains produced as 4,350 nkat/ml β-xylanase under the control of the methanol inducible alcohol oxidase 1 (AOX1 promoter. The secreted recombinant Xyn2 was estimated by SDS-PAGE to be 21 kDa. The activity of the recombinant Xyn2 was highest at 60°C and it was active over a broad range of pH (3.0–8.0 with maximal activity at pH 6.0. The enzyme was quite stable at 50°C and retained more than 94% of its activity after 30 mins incubation at this temperature. Using Birchwood xylan, the determined apparent Km and kcat values were 2.1 mg/ml and 219.2 S-1, respectively. The enzyme was highly specific towards xylan and analysis of xylan hydrolysis products confirmed as expected that the enzyme functions as endo-xylanase with xylotriose as the main hydrolysis products. The produced xylanase was practically free of cellulolytic activity. Conclusion The P. pastoris expression system allows a high level expression of xylanases. Xylanase was the main protein species in the culture supernatant, and the functional tests indicated that even the non-purified enzyme shows highly specific xylanase activity that is free of cellulolytic side acitivities. Therefore, P pastoris is a very useful expression system when the goal is highly specific and large scale production of glycosyl hydrolases.

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

    OpenAIRE

    Gilvan Caetano Duarte; Leonora Rios de Souza Moreira; Diana Paola Gómez-Mendoza; Félix Gonçalves Siqueira; Luís Roberto Batista; Lourdes Isabel Velho do Amaral; Carlos André Ornelas Ricart; Edivaldo Ximenes Ferreira Filho

    2012-01-01

    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...

  14. Production of cellulase-free xylanase by Trichoderma reesei SAF3 Produção de xilanase livre de celulase por Trichoderma reesei SAF3

    OpenAIRE

    Sanjay Kar; Asish Mandal; Das Mohapatra, Pradeep K.; Mondal, Keshab C.; Bikash R. Pati

    2006-01-01

    A xylanase producing fungi has been isolated from soil and identified as Trichoderma reesei SAF3. Maximum growth of the organism was found at 48 h under submerged condition in xylan containing enriched medium, whereas highest enzyme production (4.75U/mL) was recorded at 72 h. No detectable cellulase activity was noted during whole cultivation period. The partially purified enzyme hydrolyzed xylan into xylopentose and xylose. All these properties of xylanase highlighten its promising uses in i...

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

    Directory of Open Access Journals (Sweden)

    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.

  16. Production of xylooligosaccharides in SSF by Bacillus subtilis KCX006 producing β-xylosidase-free endo-xylanase and multiple xylan debranching enzymes.

    Science.gov (United States)

    Reddy, Shyam Sunder; Krishnan, Chandraraj

    2016-01-01

    Xylanase and xylooligosaccharides (XOS) are employed in food and feed industries. Though xylanase production from lignocellulosic materials (LCMs) by solid-state fermentation (SSF) is well known, the XOS formed during growth is not recovered due to its conversion to xylose by β-xylosidase and subsequent bacterial metabolism. A new strain, Bacillus subtilis KCX006, was exceptionally found to synthesize β-xylosidase-free endo-xylanase and multiple xylan debranching enzymes constitutively in the presence of LCMs. Absence of β-xylosidase resulted in accumulation of XOS during growth of KCX006 on LCMs. Therefore, this strain was used for simultaneous production of xylanase and XOS from agro-residues in solid-state fermentation (SSF). Partial purification of XOS from culture supernatant using activated charcoal followed by high-performance liquid chromatography (HPLC) analysis showed xylobiose to xylotetraose formed as the major products. Among various LCM substrates, wheat bran and groundnut oil-cake supported highest xylanase and XOS production at 2158 IU/gdw and 24.92 mg/gdw, respectively. The levels of xylanase and XOS were improved by 1.5-fold (3102 IU/gdw) and 1.9-fold (48 mg/gdw), respectively, by optimization of culture conditions. PMID:25310011

  17. 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.

  18. Production and characterization of cellulase-free xylanase from Trichoderma inhamatum.

    Science.gov (United States)

    de Oliveira da Silva, Leonor Alves; Carmona, Eleonora Cano

    2008-08-01

    The production of extracellular cellulase-free xylanase from Trichoderma inhamatum was evaluated in liquid Vogel medium with different carbon sources as natural substrates and agricultural or agro-industrial wastes. Optimal production of 244.02 U/mL was obtained with xylan as carbon source, pH 6.0 at 25 degrees C, 120 rpm, and 60-h time culture. Optimal conditions for enzyme activity were 50 degrees C and pH 5.5. Thermal stability of T. inhamatum xylanolytic complex expressed as T1/2 was 2.2 h at 40 degrees C and 2 min at 50 degrees C. The pH stability was high from 4.0 to 11.0. These results indicate possible employment of such enzymatic complex in some industrial processes which require activity in acid pH, wide-ranging pH stability, and cellulase activity absence. PMID:18607546

  19. 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

    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 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...... by being able to hydrolyze galactomannan (locust bean gum). Adsorption studies revealed that the smaller of the two enzymes was not able to bind to cellulose. Similarity in molecular mass, pI and hydrolytic properties suggested that these two enzymes were identical, but the smaller one was lacking...

  20. 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.

  1. EFFECT OF PRIOR MECHANICAL REFINING ON BIOBLEACHING OF WHEAT STRAW PULP WITH LACCASE /XYLANASE TREATMENT

    Directory of Open Access Journals (Sweden)

    Hai-Lan Lian,

    2012-06-01

    Full Text Available Wheat straw pulp was mechanochemically processed in a PFI mill in order to improve the effect of laccase/xylanase system (LXS treatment before bleaching. The delignification and bleachability of the prepared pulp were investigated. The delignification of the prepared pulp could be enhanced with the mechanochemical processing (refining and LXS treatment. The delignification was increased by 29.8% with refining 7000 revolutions and 5 IU/g enzyme dosage. The LXS treatment after the mechanochemical process could save 28.6% effective usage of chlorine in the subsequent hypochlorite bleaching process, compared with the traditional bio-bleaching. The crystallinity of cellulose was increased by the co-treatment with mechanochemistry and LXS treatment. This result was further supported by the observations from SEM. This co-treatment with mechanochemistry and bio-treatment enhanced the delignification and bleachability of pulp.

  2. 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.

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

    International Nuclear Information System (INIS)

    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/cm2. Gamma radiation produced new mutant strain PCGR6 which produced 26% more enzyme than the mother strain at dose 250 Gy.(author)

  4. Purification and characterization of a new Xylanase from Humicola grisea var. Thermoidea Produção, purificação e caracterização de uma nova Xilanase de Humicola grisea var. Thermoidea

    OpenAIRE

    Severino de Albuquerque Lucena-Neto; Edivaldo Ximenes Ferreira-Filho

    2004-01-01

    The thermophilic fungus Humicola grisea var. thermoidea secretes extracellular xylanase when grown on solid and in liquid media containing wheat bran and banana plant residue as substrates, respectively. At 55ºC, xylanase from the culture filtrate of H. grisea var. thermoidea grown on banana stalk retained 50% of its activity after 28 h of incubation. A xylanase (X2) was isolated from solid state cultures with wheat bran as the carbon source. It was purified to apparent homogeneity by ultrafi...

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

    Directory of Open Access Journals (Sweden)

    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.

  6. Production of xylan degrading endo-1, 4-β-xylanase from thermophilic Geobacillus stearothermophilus KIBGE-IB29

    Directory of Open Access Journals (Sweden)

    Zainab Bibi

    2014-10-01

    Full Text Available Xylan degrading bacterial strain was isolated from soil and identified as Geobacillus stearothermophilus KIBGE-IB29 on the basis of morphological, biochemical and 16S rDNA sequence analysis. Optimization of medium and culture conditions in submerged fermentation was investigated for maximum endo-1, 4-β-xylanase production. High yield of xylan degrading endo-1, 4-β-xylanase was achieved at 60 °C and pH-6.0 with 24 h of fermentation. Maximum enzyme was produced using 0.5% xylan as a carbon source, 0.5% peptone, 0.2% yeast extract and 0.1% meat extract as nitrogen sources. Di-potassium hydrogen phosphate (0.25%, calcium chloride (0.01%, potassium hydrogen phosphate (0.05% and ammonium sulfate (0.05% were also incorporated in the fermentation medium to enhance the enzyme production.

  7. Partial purification and properties of cellulase-free alkaline xylanase produced by Rhizopus stolonifer in solid-state fermentation

    OpenAIRE

    Antonio José Goulart; Eleonora Cano Carmona; Rubens Monti

    2005-01-01

    Rhizopus stolonifer was cultivated in wheat bran to produce a cellulase-free alkaline xylanase. The purified enzyme obtained after molecular exclusion chromatography in Sephacryl S-200 HR showed optimum temperature as 45º C and hydrolysis pHs optima as pH 6.0 and 9.0. Xylanase presented higher Vmax at pH 9.0 (0.87 µmol/mg protein) than at pH 6.0 and minor Km at pH 6.0 (7.42 mg/mL) than at pH 9.0.Rhizopus stolonifer foi cultivado em meio de farelo de trigo para produzir uma xilanase alcalina c...

  8. 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.

  9. Xylanase and β-xylosidase production by Aspergillus ochraceus : new perspectives for the application of wheat straw autohydrolysis liquor

    OpenAIRE

    Michelin, Michele; Maria de Lourdes T. M Polizeli; Ruzene, Denise S.; Silva, Daniel Pereira da; Vicente, A.A.; Jorge, João A.; Terenzi, Héctor F.; Teixeira, J.A.

    2012-01-01

    The xylanase biosynthesis is induced by its substrate—xylan. The high xylan content in some wastes such as wheat residues (wheat bran and wheat straw) makes them accessible and cheap sources of inducers to be mainly applied in great volumes of fermentation, such as those of industrial bioreactors. Thus, in this work, the main proposal was incorporated in the nutrient medium wheat straw particles decomposed to soluble compounds (liquor) through treatment of lignocellulosic materials in autohyd...

  10. 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.

  11. Characterization of a purified thermostable xylanase from Caldicoprobacter algeriensis sp. nov. strain TH7C1(T).

    Science.gov (United States)

    Amel, Bouanane-Darenfed; Nawel, Boucherba; Khelifa, Bouacem; Mohammed, Gagaoua; Manon, Joseph; Salima, Kebbouche-Gana; Farida, Nateche; Hocine, Hacene; Bernard, Ollivier; Jean-Luc, Cayol; Marie-Laure, Fardeau

    2016-01-01

    The present study investigates the purification and biochemical characterization of an extracellular thermostable xylanase (called XYN35) from Caldicoprobacter algeriensis sp. nov., strain TH7C1(T), a thermophilic, anaerobic strain isolated from the hydrothermal hot spring of Guelma (Algeria). The maximum xylanase activity recorded after 24 h of incubation at 70 °C and in an optimized medium containing 10 g/L mix birchwood- and oats spelt-xylan was 250 U/mL. The pure protein was obtained after heat treatment (1 h at 70 °C), followed by sequential column chromatographies on Sephacryl S-200 gel filtration and Mono-S Sepharose anion-exchange. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis indicated that the purified enzyme is a monomer with a molecular mass of 35,075.10 Da. The results from amino-acid sequence analysis revealed high homology between the 21 NH2-terminal residues of XYN35 and those of bacterial xylanases. The enzyme showed optimum activity at pH 11 and 70 °C. While XYN35 was activated by Ca(2+), Mn(2+), and Mg(2+), it was completely inhibited by Hg(2+) and Cd(2+). The xylanase showed higher specific activity on soluble oat-spelt xylan, followed by beechwood xylan. This enzyme was also noted to obey the Michaelis-Menten kinetics, with Km and kcat values on oat-spelt xylan being 1.33 mg/mL and 400 min(-1), respectively. Thin-layer chromatography soluble oat-spelt xylan (TLC) analysis showed that the final hydrolyzed products of the enzyme from birchwood xylan were xylose, xylobiose, and xylotriose. Taken together, the results indicated that the XYN35 enzyme has a number of attractive biochemical properties that make it a potential promising candidate for future application in the pulp bleaching industry. PMID:26687892

  12. Cloning, Expression and Characteristics of a Novel Alkalistable and Thermostable Xylanase Encoding Gene (Mxyl) Retrieved from Compost-Soil Metagenome

    OpenAIRE

    Verma, Digvijay; Kawarabayasi, Yutaka; Miyazaki, Kentaro; Satyanarayana, Tulasi

    2013-01-01

    Background The alkalistable and thermostable xylanases are in high demand for pulp bleaching in paper industry and generating xylooligosaccharides by hydrolyzing xylan component of agro-residues. The compost-soil samples, one of the hot environments, are expected to be a rich source of microbes with thermostable enzymes. Methodology/Principal Findings Metagenomic DNA from hot environmental samples could be a rich source of novel biocatalysts. While screening metagenomic library constructed fr...

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

    OpenAIRE

    J Davoodi; 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...

  14. STATISTICAL OPTIMIZATION OF MINERAL SALT AND UREA CONCENTRATION FOR CELLULASE AND XYLANASE PRODUCTION BY Penicillium echinulatum IN SUBMERGED FERMENTATION

    Directory of Open Access Journals (Sweden)

    L. dos Reis

    2015-03-01

    Full Text Available Abstract Penicillium echinulatum S1M29 is a mutant with cellulase and xylanase production comparable to the most studied microorganisms in the literature. However, its potential to produce these enzymes has not been fully investigated. This study aimed at optimizing salt and urea concentrations in the mineral solution, employing the response surface methodology. A 25-1 Fractional Factorial Design and a 23 Central Composite Design were applied to elucidate the effect of salts and urea in enzyme production. Lower concentrations of KH2PO4 (2.0 g.L-1, (NH42SO4 (1.4 g.L-1, MgSO4.7H2O (0.375 g.L-1 and CaCl2 (0.375 g.L-1 were most suitable for the production of all enzymes evaluated. Nevertheless, higher concentrations of urea (0.525 g.L-1 gave the best results for cellulase and xylanase production. The maximum FPase (1,5 U.m.L-1, endoglucanase (7,2 U.m.L-1, xylanase (30,5 U.m.L-1 and β-glucosidase (4,0 U.m.L-1 activities obtained with the planned medium were, respectively, 87, 16, 17 and 21% higher when compared to standard medium. The experimental design contributed to adjust the concentrations of minerals and urea of the culture media for cellulase and xylanase production by P. echinulatum, avoiding waste of components in the medium.

  15. CLONING, EXPRESSION, AND CHARACTERIZATION OF AN ALKALOPHILLIC ENDO-1,4-BETA-XYLANASE FROM PAENIBACILLUS SP. HPL-002

    Directory of Open Access Journals (Sweden)

    No-Joong Park,

    2011-12-01

    Full Text Available The biochemical properties of a purified enzyme of a new alkalophillic endo-1,4-beta-xylanase gene, KRICT PX2 (GU967374, which was isolated from Paenibacillus sp. HPL-002 (KCTC11410BP and expressed in E. coli, were investigated. The specific activity of the purified xylanase was 51.26 μmol/min/mg proteins. The Km and Vmax values of the protein for birch wood xylan were also verified to have 0.061 μM and 55.3 μmol/min/mg proteins, respectively. The optimum pH and temperature for the activity of the enzyme were pH 8~9 and 50oC, respectively, and, the activity was stably maintained at 40oC. Most metallic salts, ethylenediamine tetra-acetic acid, 2-mercaptoethanol, phenylmethane-sulphonyl fluoride, and furfural, have no impact on the enzyme’s activity at 1 mM. The simulated 3-D structure of this xylanase is similar to Xyn10B from Paenibacillus barcinonensis. Further research on the degradation of different-origin xylans and enzyme production will be necessary for practical applications.

  16. A xylanase from Streptomyces sp. FA1: heterologous expression, characterization, and its application in Chinese steamed bread.

    Science.gov (United States)

    Xu, Yang; Wu, Jing; Zheng, Kaixuan; Wu, Dan

    2016-05-01

    Xylanases (EC 3.2.1.8) are hydrolytic enzymes that have found widespread application in the food, feed, and paper-pulp industries. Streptomyces sp. FA1 xynA was expressed as a secreted protein in Pichia pastoris, and the xylanase was applied to the production of Chinese steamed bread for the first time. The optimal pH and the optimal temperature of XynA were 5.5 and 60 °C, respectively. Using beechwood as substrate, the K m and V max were 2.408 mg mL(-1) and 299.3 µmol min(-1) mg(-1), respectively. Under optimal conditions, a 3.6-L bioreactor produced 1374 U mL(-1) of XynA activity at a protein concentration of 6.3 g L(-1) after 132 h of fermentation. Use of recombinant XynA led to a greater increase in the specific volume of the CSB than could be achieved using commercial xylanase under optimal conditions. This study provides the basis for the application of the enzyme in the baking industry. PMID:26803505

  17. Cloning and expression of a novel, moderately thermostable xylanase-encoding gene (Cflxyn11A) from Cellulomonas flavigena.

    Science.gov (United States)

    Amaya-Delgado, Lorena; Mejía-Castillo, Teresa; Santiago-Hernández, Alejandro; Vega-Estrada, Jesús; Amelia, Farrés-G-S; Xoconostle-Cázares, Beatriz; Ruiz-Medrano, Roberto; Montes-Horcasitas, María Del Carmen; Hidalgo-Lara, María Eugenia

    2010-07-01

    The Cfl xyn11A gene, encoding the endo-1,4-beta-xylanase Cfl Xyn11A from Cellulomonas flavigena, was isolated from a genomic DNA library. The open reading frame of the Cfl xyn11A gene was 999 base pairs long and encoded a polypeptide (Cfl Xyn11A) of 332 amino acids with a calculated molecular mass of 35,110Da. The Cfl xyn11A gene was expressed in Escherichia coli and the recombinant enzyme, with an estimated molecular weight of 31kDa was purified and xylanase activity was measured. Cfl Xyn11A showed optimal activity at pH 6.5 and 55 degrees C. The enzyme demonstrated moderate thermal stability as Cfl Xyn11A maintained 50% of its activity when incubated at 55 degrees C for 1h or at 45 degrees C for 6h. This is the first report describing the cloning, expression and functional characterization of an endo-1,4-beta-xylanase-encoding gene from C. flavigena. Cfl Xyn11A may be suitable for industrial applications in the food and feed industries, or in the pre-treatment of lignocellulosic biomass required to improve the yields of fermentable sugars for bioethanol production. PMID:20231092

  18. Expression of an alkalo-tolerant fungal xylanase enhanced by directed evolution in Pichia pastoris and Escherichia coli.

    Science.gov (United States)

    McHunu, Nokuthula Peace; Singh, Suren; Permaul, Kugen

    2009-04-20

    The alkaline stability of the xylanase from Thermomyces lanuginosus was further improved by directed evolution using error-prone PCR mutagenesis. Positive clones were selected by their ability to produce zones of clearing on pH 9 and 12 xylan agar plates. Variant NC38 was able to withstand harsh alkaline conditions retaining 84% activity after exposure at pH 10 for 90 min at 60 degrees C, while the parent enzyme had 22% activity after 60 min. The alkaline stable variant NC38 was cloned into pBGP1 under the control GAP promoter and pET22b(+) for expression in Pichia pastoris and Escherichia coli BL21, respectively. Best extracellular expression of the recombinant xylanase was observed in P. pastoris (261.7+/-0.61 U ml(-1)) whereas intracellular activity was observed in E. coli (47.9+/-0.28 U ml(-1)) was low. Total activity obtained in P. pastoris was 545-fold higher than E. coli. The mutated alkaline stable xylanase from P. pastoris was secreted into the culture medium with little or no contamination by host proteins, which favours the application of this enzyme in the pulp and paper industry. PMID:19428727

  19. 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.

  20. 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.

  1. Recombination of thermo-alkalistable, high xylooligosaccharides producing endo-xylanase from Thermobifida fusca and expression in Pichia pastoris.

    Science.gov (United States)

    Wang, Qian; Du, Wen; Weng, Xiao-Yan; Liu, Ming-Qi; Wang, Jia-Kun; Liu, Jian-Xin

    2015-02-01

    For xylooligosaccharide (XO) production, endo-xylanase from Thermobifida fusca was modified by error-prone PCR and DNA shuffling. The G4SM1 mutant (S62T, S144C, N198D, and A217V) showed the most improved hydrolytic activity and was two copies expressed in Pichia pastoris under the control of GAP promoter. The maximum xylanase activity in culture supernatants was 165 ± 5.5 U/ml, and the secreted protein concentration reached 493 mg/l in a 2-l baffled shake flask. After 6× His-tagged protein purification, the specific activity of G4SM1 was 2036 ± 45.8 U/mg, 2.12 times greater than that of wild-type enzyme. Additionally, G4SM1 was stable over a wide pH range from 5.0 to 9.0. Meanwhile, half-life of G4SM1 thermal inactivation at 70 °C increased 8.5-fold. Three-dimensional structures suggest that two amino acid substitutions, S62T and S144C, located at catalytic domain may be responsible for the enhanced activity and thermostability of xylanase. Xylobiose was the dominant end product of xylan hydrolysis by G4SM1. Due to its attractive biochemical properties, G4SM1 has potential value in commercial XO production. PMID:25384545

  2. High-level expression and characterization of a thermostable xylanase mutant from Trichoderma reesei in Pichia pastoris.

    Science.gov (United States)

    Li, Yang-yuan; Zhong, Kai-xin; Hu, Ai-hong; Liu, Dan-ni; Chen, Li-zhi; Xu, Shu-de

    2015-04-01

    A gene encoding xylanase 2 mutant from Trichoderma reesei (T2C/T28C, named mxyn2) was cloned into the Pichia pastoris X33 strain using the vector pPICZαA. Recombinant Mxyn2p was functionally expressed in P. pastoris X33 and secreted into the supernatant. Real time qPCR demonstrated that an increase in gene copy number correlated with higher levels of expression. Supernatant from methanol induced cells was concentrated by ultrafiltration with a 10kDa cut off membrane, and purified with ion exchange chromatography using SP Sepharose Fast Flow chromatography. Recombinant Mxyn2p protein had the highest activity at 75°C, while recombinant protein encoded by the "wild type" xylanase gene xyn2, also expressed in Pichia, was 20°C lower. The Mxyn2p enzyme retained more than 70% of its activity after incubation at 80°C for 10min. The effects of the optimal pH and temperature for higher expression levels in P. pastoris were also determined, 6.0 and 22°C, respectively. The maximum xylanase activity of Mxyn2p was 13,000nkat/mg (9.88g/l) in fed-batch cultivation after 168h induction with methanol in a 50l bioreactor. PMID:25434687

  3. A novel low molecular weight endo-xylanase from Streptomyces sp. CS628 cultivated in wheat bran.

    Science.gov (United States)

    Rahman, Md Arifur; Choi, Yun Hee; Pradeep, G C; Choi, Yoon Seok; Choi, Eun Joo; Cho, Seung Sik; Yoo, Jin Cheol

    2014-07-01

    An extracellular low molecular weight xylanase (Xyn628) from Streptomyces sp. CS628 was isolated from Korean soil sample, produced in wheat bran medium, purified, and biochemically characterized. Xyn628 was purified 4.8-fold with a 33.78 % yield using Sepharose CL-6B column chromatography. The purified xylanase was ~18.1 kDa estimated by SDS-PAGE and xylan zymography. N-terminal amino acid sequences of Xyn628 were AYIKEVVSRAYM. The enzyme was found to be stable in a broad range of pH (5.0-13.0) and up to 60 °C and have optimal pH and temperature of pH 11.0 and 60 °C, respectively. Xyn628 activities were remarkable affected by various detergents, chelators, modulators, and metal ions. The xylanase produced xylobiose and xylotriose as principal hydrolyzed end products from the xylan. It was found to degrade agro-waste materials like corn cob and wheat bran by Xyn628 (20 U/g) as shown by electron microscopy. As being simple in purification, low molecular weight, alkaline, thermostable, and ability to produce xylooligosaccharides show that Xyn628 has potential applications in bioindustries as a biobleaching agent or/and xylooligosaccharides production with an appropriate utilization of agro-waste. PMID:24817510

  4. Determination of the modes of action and synergies of xylanases by analysis of xylooligosaccharide profiles over time using fluorescence-assisted carbohydrate electrophoresis.

    Science.gov (United States)

    Gong, Weili; Zhang, Huaiqiang; Tian, Li; Liu, Shijia; Wu, Xiuyun; Li, Fuli; Wang, Lushan

    2016-07-01

    The structure of xylan, which has a 1,4-linked β-xylose backbone with various substituents, is much more heterogeneous and complex than that of cellulose. Because of this, complete degradation of xylan needs a large number of enzymes that includes GH10, GH11, and GH3 family xylanases together with auxiliary enzymes. Fluorescence-assisted carbohydrate electrophoresis (FACE) is able to accurately differentiate unsubstituted and substituted xylooligosaccharides (XOS) in the heterogeneous products generated by different xylanases and allows changes in concentrations of specific XOS to be analyzed quantitatively. Based on a quantitative analysis of XOS profiles over time using FACE, we have demonstrated that GH10 and GH11 family xylanases immediately degrade xylan into sizeable XOS, which are converted into smaller XOS in a much lower speed. The shortest substituted XOS produced by hydrolysis of the substituted xylan backbone by GH10 and GH11 family xylanases were MeGlcA(2) Xyl3 and MeGlcA(2) Xyl4 , respectively. The unsubstituted xylan backbone was degraded into xylose, xylobiose, and xylotriose by both GH10 and GH11 family xylanases; the product profiles are not family-specific but, instead, depend on different subsite binding affinities in the active sites of individual enzymes. Synergystic action between xylanases and β-xylosidase degraded MeGlcA(2) Xyl4 into xylose and MeGlcA(2) Xyl3 but further degradation of MeGlcA(2) Xyl3 required additional enzymes. Synergy between xylanases and β-xylosidase was also found to significantly accelerate the conversion of XOS into xylose. PMID:27060349

  5. Decolorization of four dyes by a mixed culture of Trametes sp. SQO1 and Chaetomium sp. RO1%Trametes sp.SQ01和Chaetomium sp.R01混合培养对四种染料的脱色

    Institute of Scientific and Technical Information of China (English)

    杨秀清; 王婧人; 赵晓霞; 薛瑞

    2011-01-01

    Four dyes including Congo red, acid red, orange G and bromphenol blue were decolorized by a mixed fungal culture of Trametes sp. SQO 1 and Chaetomium sp. R0I, which were newly developed in our laboratory. The results indicated that MnP activity in the mixed culture of strain S001 and R0I increased approximately 5.5 times over that obtained from a pure culture of SQOI. The amount and timing of strain R01 had a marked effect on MnP production by SQOI. If the inoculum of strain ROI was too high or too low, the MnP activity decreased in the mixed culture. As the inoculation time of strain R01 was delayed, MnP activity was gradually reduced. MnP production was inhibited by about 40% ~75% by the four dyes in mixed culture of SQOI and R01. The decolorization rates of mixed culture of strain SQ01 and R01 were increased by 20% ~ 50% compared to that of the monoculture of SQOI. After 3 days, 75% ~94% of four dyes were decolorized by mixed culture of SQ01 and R01 ,while the decolorization rate was only about 41% 75% by strain SQ01 cultured alone. The time-point of dye addition remarkably influenced the efficiency of decolorization by the mixed culture. The decolorization rates of dyes added to the medium after 4 days of mixed cultivation were significantly higher as compared to dyes added at the outset of mixed cultivation. In addition to biodegradation, biosorption also played an important role in the decoiorization process. The biosorption of orange G, bromphenol blue and acid red accounted for 2% ,5% and 15% of the total color removal,respectively. However,the biosorption rate of congo red reached up to 60%.To date, this is the first report of decolorizing dyes by mixed fungal cultures. The high efficiency of the decolorization ability indicates that the mixed culture has broad application prospects in dye decolorization treatment.%利用本实验室新构建的白腐菌Trametes sp.SQ01和毛壳菌Chaetomium sp.R01混合培养体系,对刚

  6. Cloning and constitutive expression of His-tagged xylanase GH 11 from Penicillium occitanis Pol6 in Pichia pastoris X33: purification and characterization.

    Science.gov (United States)

    Driss, Dorra; Bhiri, Fatma; Ghorbel, Raoudha; Chaabouni, Semia Ellouz

    2012-05-01

    High-level constitutive expression of xylanase GH11 from Penicillium occitanis Pol6 termed PoXyn2 was achieved using the methylotrophic yeast Pichia pastoris. The PoXyn2 cDNA encoding for a mature xylanase of 320 amino acids was subcloned into the pGAPZαA vector, to construct recombinant xylanse with six histidine residues at the N-terminal and further integrated into the genome of P. pastoris X-33 under the control of the glyceraldehyde 3-phosphate dehydrogenase (GAP) constitutive promoter. Activity assay and SDS-PAGE demonstrate that the His-tagged xylanase was extracellularly expressed in P. pastoris and purified to homogeneity by a simple, one-step purification protocol using immobilized metal affinity chromatography (Ni-NTA resin). The purified PoXyn2 showed a single band on SDS-PAGE with an apparent molecular weight of 30 kDa. The xylanase activity was optimal at pH 3.0 and 50°C. The specific activity measured for Oat Spelt Xylan was 8549.85 U mg(-1). The apparent The K(M) and V(max) values were 8.33±0.7 mg ml(-1)and 58.82±0.9 μmol min(-1) ml(-1), respectively, as measured on Oat Spelt Xylan. This is the first report demonstrating the possibility of mass production of P. occitanis xylanase using P. pastoris. PMID:22402470

  7. Purification and characterization of cellulase-free low molecular weight endo β-1,4 xylanase from an alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost.

    Science.gov (United States)

    Walia, Abhishek; Mehta, Preeti; Chauhan, Anjali; Kulshrestha, Saurabh; Shirkot, C K

    2014-10-01

    Alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost is first report on actinomycetes that has the ability to produce thermostable cellulase-free xylanase, which is an important industrial enzyme used in the pulp and paper industry. Strain CKMX1 was characterized by metabolic fingerprinting, whole-cell fatty acids methyl ester analysis and 16Sr DNA and found to be C. cellulans CKMX1.The enzyme was purified by gel permeation and anion exchange chromatography and had a molecular mass of 29 kDa. Xylanase activity was optimum at pH 8.0 and 55 °C. The enzyme was somewhat thermostable, retaining 50 % of the original activity after incubation at 50 °C for 30 min. The xylanase had K m and V max values of 2.64 mg/ml and 2,000 µmol/min/mg protein in oat spelt xylan, respectively. All metal ions except HgCl2, CoCl2 as well as CdCl2 were well tolerated and did not adversely affect xylanase activity. The deduced internal amino acid sequence of C. cellulans CKMX1 xylanase by matrix assisted laser desorption ionization-time of flight mass spectrometry resembled the sequence of β-1,4-endoxylanase, which is a member of glycoside hydrolase family 11. Some of the novel characteristics that make this enzyme potentially effective in xylan biodegradation could be useful for pulp and paper biobleaching are discussed in this manuscript. PMID:24908422

  8. Ethanol from a biorefinery waste stream: Saccharification of amylase, protease and xylanase treated wheat bran.

    Science.gov (United States)

    Wood, Ian P; Cook, Nicola M; Wilson, David R; Ryden, Peter; Robertson, James A; Waldron, Keith W

    2016-05-01

    Biorefining aims to exploit the full value of plant material by sequentially extracting and valorising its components. Many studies focus on the saccharification of virgin biomass sources, but it may be more efficient to pre-extract high-value components before hydrolysis to fermentable sugars. In the current study, a bran residue from de-starched, protein depleted and xylanase treated wheat bran has been subjected to hydrothermal pretreatment, saccharification and fermentation procedures to convert the residue to ethanol. The most effective pretreatment conditions (>190 °C, 10 min) and saccharification conditions were identified following bench-scale liquid hot water pretreatment. Pre-extraction of enzymatically-hydrolysable starch and xylan reduced the release of furfural production, particularly when lower pretreatment severities were used. Pilot-scale steam explosion of the lignocellulosic residue followed by cellulase treatment and conversion to ethanol at a high substrate concentration (19%) gave an ethanol titre of ≈ 25 g/L or a yield of 93% of the theoretical maximum. PMID:26769514

  9. 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.

  10. Expression of A. niger US368 xylanase in E. coli: purification, characterization and copper activation.

    Science.gov (United States)

    Elgharbi, Fatma; Hlima, Hajer Ben; Farhat-Khemakhem, Ameny; Ayadi-Zouari, Dorra; Bejar, Samir; Hmida-Sayari, Aïda

    2015-03-01

    The XAn11 cDNA was cloned in pET-28a(+) and the recombinant plasmid was transformed in Escherichia coli. The His-tagged r-XAn11 was purified using Ni-NTA affinity and anion exchange chromatography. The enzyme showed a specific activity of 415.1 U mg(-1) and a molecular mass of 25 kDa. It had an optimal activity at pH 5 and 50°C. It was stable in a wide range of pH and in the presence of some detergents and organic solvents. In the presence of 3mM Cu2+, the relative activity of the His-tagged r-XAn11 was enhanced by 54%. This is the first work reporting that copper is a strong activator for xylanase activity making this enzyme very attractive for future industrial applications. Molecular modeling suggests that the contact region between the catalytic site and the N-terminal His-tag fusion peptide could be responsible for the different behavior of the native and recombinant enzyme toward copper. PMID:25530001

  11. Large conformational fluctuations of the multi-domain xylanase Z of Clostridium thermocellum.

    Science.gov (United States)

    Różycki, Bartosz; Cieplak, Marek; Czjzek, Mirjam

    2015-07-01

    The cellulosome is a multi-enzyme machinery which efficiently degrades plant cell-wall polysaccharides. The multiple domains of the cellulosome complexes are often tethered to one another by intrinsically disordered regions. The properties and functions of these disordered linkers are unknown to a large extent. In this work, we study the conformational variability of one component of the cellulosome - the multi-domain xylanase Z (XynZ) of Clostridium thermocellum. We use a coarse-grained protein model to efficiently simulate conformations of the enzyme. Our simulation results are in excellent agreement with data from small angle X-ray scattering experiments, which validates the simulation outcome. Both in the presence and absence of the cohesin domain, the XynZ enzyme appears to be flexible in the sense that it takes various compact and extended conformations. The physical interactions between the individual domains are rather weak and transient, and the XynZ enzyme is held together mainly by the flexible linkers connecting the domains. The end-to-end distance distributions for the flexible linkers can be rationalized by the excluded volume effect. Taken together, our results provide a detailed picture of the conformational ensemble of the XynZ enzyme in solution. PMID:26008791

  12. Xylanase supplementation of a wheat-based diet improved nutrient digestion and mRNA expression of intestinal nutrient transporters in broiler chickens infected with Clostridium perfringens.

    Science.gov (United States)

    Guo, Shuangshuang; Liu, Dan; Zhao, Xu; Li, Changwu; Guo, Yuming

    2014-01-01

    Necrotic enteritis caused by Clostridium perfringens has become prevalent in the European Union due to the withdrawal of antibiotics in poultry feed. In an experiment with a 2 × 2 factorial arrangement, 336 one-day-old male broiler chicks (Ross 308) were assigned to 4 groups with or without C. perfringens challenge and fed wheat-based diets supplemented with or without xylanase at 5,500 U/kg of diet. The study aimed to investigate effects of xylanase addition on growth performance as well as nutrient digestion and absorption of C. perfringens-infected broilers. Before challenge (d 0-14), xylanase-supplemented birds had greater ADG and lower feed conversion ratio (FCR; P Clostridium perfringens infection decreased AME values and apparent ileal digestibility of DM of diets (P perfringens infection (P perfringens infection and elevated apparent ileal digestibility of CP and mRNA expression of nutrient transporters in challenged birds. PMID:24570428

  13. Crystallization and preliminary X-ray analysis of a cold-active endo-β-1,4-d-xylanase from glycoside hydrolase family 8

    International Nuclear Information System (INIS)

    The crystallization and preliminary X-ray analysis of a cold-active endo-β-1,4-d-xylanase is described. The crystals diffracted to 2.7 Å resolution. Endo-β-1,4-d-xylanases are used in a multitude of industrial applications. Native crystals of a cold-adapted xylanase from glycoside hydrolase family 8 were obtained by the vapour-diffusion technique. The crystals belonged to space group I222, with unit-cell parameters a = 46.6, b = 110.8, c = 150.2 Å at 100 K, and diffracted to 2.7 Å resolution at a synchrotron source. The asymmetric unit is likely to contain one molecule, with a VM of 2.07 Å3 Da−1, corresponding to a solvent content of ∼40%

  14. Phylogenetic analysis of β-xylanase SRXL1 of Sporisorium reilianum and its relationship with families (GH10 and GH11) of Ascomycetes and Basidiomycetes

    Science.gov (United States)

    Álvarez-Cervantes, Jorge; Díaz-Godínez, Gerardo; Mercado-Flores, Yuridia; Gupta, Vijai Kumar; Anducho-Reyes, Miguel Angel

    2016-01-01

    In this paper, the amino acid sequence of the β-xylanase SRXL1 of Sporisorium reilianum, which is a pathogenic fungus of maize was used as a model protein to find its phylogenetic relationship with other xylanases of Ascomycetes and Basidiomycetes and the information obtained allowed to establish a hypothesis of monophyly and of biological role. 84 amino acid sequences of β-xylanase obtained from the GenBank database was used. Groupings analysis of higher-level in the Pfam database allowed to determine that the proteins under study were classified into the GH10 and GH11 families, based on the regions of highly conserved amino acids, 233–318 and 180–193 respectively, where glutamate residues are responsible for the catalysis. PMID:27040368

  15. 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.

  16. Effect of polyols on thermostability of xylanase from a tropical isolate of Aureobasidium pullulans and its application in prebleaching of rice straw pulp

    OpenAIRE

    Bankeeree, Wichanee; Lotrakul, Pongtharin; Prasongsuk, Sehanat; Chaiareekij, Somporn; Eveleigh, Douglas E.; Kim, Seung Wook; Punnapayak, Hunsa

    2014-01-01

    In an attempt to find a thermostable xylanase enzyme for potential application in the pretreatment prior to H2O2 bleaching of paper pulp for industry, an extracellular xylanase from Aureobasidium pullulans CBS 135684 was purified 17.3-fold to apparent homogeneity with a recovery yield of 13.7%. Its molecular mass was approximately 72 kDa as determined by SDS-PAGE. The optimal pH and temperature for activity of the purified enzyme were pH 6.0 and 70°C, respectively. The enzyme was relatively s...

  17. Heterologous Expression of Family 10 Xylanases from Acidothermus cellulolyticus Enhances the Exoproteome of Caldicellulosiruptor bescii and Growth on Xylan Substrates

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sun-Ki; Chung, Daehwan; Himmel, Michael E.; Bomble, Yannick J.; Westpheling, Janet

    2016-08-22

    The ability to deconstruct plant biomass without conventional pretreatment has made members of the genus Caldicellulosiruptor the target of investigation for the consolidated processing of lignocellulosic biomass to biofuels and bioproducts. These Gram-positive bacteria are hyperthermophilic anaerobes and the most thermophilic cellulolytic organisms so far described. They use both C5 and C6 sugars simultaneously and have the ability to grow well on xylan, a major component of plant cell walls. This is an important advantage for their use to efficiently convert biomass at yields sufficient for an industrial process. For commodity chemicals, yield from substrate is perhaps the most important economic factor. In an attempt to improve even further the ability of C. bescii to use xylan, we introduced two xylanases from Acidothermus cellulolyticus. Acel_0180 includes tandem carbohydrate-binding modules (CBM2 and CBM3) located at the C-terminus, one of which, CBM2, is not present in C. bescii. Also, the sequences of Xyn10A and Acel_0180 have very little homology with the GH10 domains present in C. bescii. For these reasons, we selected these xylanases as potential candidates for synergistic interaction with those in the C. bescii exoproteome. Heterologous expression of two xylanases from Acidothermus cellulolyticus in Caldicellulosiruptor bescii resulted in a modest, but significant increase in the activity of the exoproteome of C. bescii on xylan substrates. Even though the increase in extracellular activity was modest, the ability of C. bescii to grow on these substrates was dramatically improved suggesting that the xylan substrate/microbe interaction substantially increased deconstruction over the secreted free enzymes alone. We anticipate that the ability to efficiently use xylan, a major component of plant cell walls for conversion of plant biomass to products of interest, will allow the conversion of renewable, sustainable, and inexpensive plant feedstocks to

  18. Enzymatic saccharification of sugar cane bagasse by continuous xylanase and cellulase production from cellulomonas flavigena PR-22.

    Science.gov (United States)

    Rojas-Rejón, Óscar A; Poggi-Varaldo, Héctor M; Ramos-Valdivia, Ana C; Ponce-Noyola, Teresa; Cristiani-Urbina, Eliseo; Martínez, Alfredo; de la Torre, Mayra

    2016-03-01

    Cellulase (CMCase) and xylanase enzyme production and saccharification of sugar cane bagasse were coupled into two stages and named enzyme production and sugar cane bagasse saccharification. The performance of Cellulomonas flavigena (Cf) PR-22 cultured in a bubble column reactor (BCR) was compared to that in a stirred tank reactor (STR). Cells cultured in the BCR presented higher yields and productivity of both CMCase and xylanase activities than those grown in the STR configuration. A continuous culture with Cf PR-22 was run in the BCR using 1% alkali-pretreated sugar cane bagasse and mineral media, at dilution rates ranging from 0.04 to 0.22 1/h. The highest enzymatic productivity values were found at 0.08 1/h with 1846.4 ± 126.4 and 101.6 ± 5.6 U/L·h for xylanase and CMCase, respectively. Effluent from the BCR in steady state was transferred to an enzymatic reactor operated in fed-batch mode with an initial load of 75 g of pretreated sugar cane bagasse; saccharification was then performed in an STR at 55°C and 300 rpm for 90 h. The constant addition of fresh enzyme as well as the increase in time of contact with the substrate increased the total soluble sugar concentration 83% compared to the value obtained in a batch enzymatic reactor. This advantageous strategy may be used for industrial enzyme pretreatment and saccharification of lignocellulosic wastes to be used in bioethanol and chemicals production from lignocellulose. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:321-326, 2016. PMID:26701152

  19. Xylanase production by Streptomyces viridosporus T7A in submerged and solid-state fermentation using agro-industrial residues

    OpenAIRE

    Luiz Romulo Alberton; Luciana Porto de Souza Vandenberghe; Ricardo Assmann; Ricardo Cancio Fendrich; José Rodriguéz-León; Carlos Ricardo Soccol

    2009-01-01

    The study of xylanase production was conducted by Streptomyces viridosporus T7A in submerged (SmF) and solid-state fermentation (SSF), using agro-industrial residues and sub-products. Napier grass, sugarcane bagasse and soybean bran were used as carbon source, substrate/support, and nitrogen source, respectively. In SmF, Napier grass (1% v/w) supplemented with soybean bran, hydroxyethylcellulose and B complex vitamins were used. Soybean bran (1.5 % w/v), B complex vitamins (0.1%), and hydroxy...

  20. Effect of temperature on the production of cellulases, xylanases and lytic enzymes by selected Trichoderma reesei mutants

    OpenAIRE

    Piotr Janas; Zdzisław Targoński

    2014-01-01

    The effect of temperature in the rangę of 26-38°C on the production of cellulases, xylanases and lytic enzymes by four mutant strains of Trichoderma reesei was analysed. On the basis of these investigations three thermosensitive strains (M-7. RUT C 30 and VTT-D-78085) which showed reduced excretion of the above mentioned enzymes as well as protein and a thermoresistant mutant (VTT-D-79I24) which grew within a temperature range of 26-34°C were characterized. Higher temperature caused an increa...

  1. Purification and characterization of a new xylanase (APX-II) from the fungus Aureobasidium pullulans Y-2311-1.

    OpenAIRE

    Li, X.L.; Z. Q. Zhang; Dean, J F; Eriksson, K E; Ljungdahl, L G

    1993-01-01

    Aureobasidium pullulans Y-2311-1 produced four major xylanases (EC 3.2.1.8) with pI values of 4.0, 7.3, 7.9, and 9.4 as revealed by isoelectric focusing and zymogram analysis when grown for 4 days on 1.0% oat spelt xylan. The enzyme with a pI of 9.4 was purified by ammonium sulfate precipitation, chromatography on a DEAE-Sephadex A-50 column, and gel filtration with a Sephadex G-75 column. The enzyme had a mass of about 25 kDa as determined by both sodium dodecyl sulfate-polyacrylamide gel el...

  2. Effect of Temperature on Xylanase II from Trichoderma reesei QM 9414: A Calorimetric, Catalytic, and Conformational Study

    OpenAIRE

    Gloria López; Pilar Estrada

    2014-01-01

    The secondary structure of xylanase II from Trichoderma reesei is lost in an apparent irreversible cooperative process as temperature is increased with a midpoint transition of 58.8 ± 0.1°C. The shift of the spectral centre of mass above 50°C is also apparently cooperative with midpoint transition of 56.3 ± 0.2°C, but the existence of two isofluorescent points in the fluorescence emission spectra suggests a non-two-state process. Further corroboration comes from differential scanning calorime...

  3. 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

  4. Behavior and supportive evidence of a large xylanase-containing multienzyme complex of Tepidimicrobium xylanilyticum BT14

    Directory of Open Access Journals (Sweden)

    Paripok Phitsuwan

    2012-11-01

    Full Text Available Cellular behaviors and a xylanolytic-cellulolytic enzyme system of Tepidimicrobium xylanilyticum BT14 towards xylan-rich plant biomass degradation were characterized. During the exponential growth phase, the bacterial cells were bound tightly to the growth substrate where the degradation zones appeared mostly around the cells, indicating that the xylanolytic-cellulolytic enzyme system was linked to the cell surfaces. Interestingly, several cells appeared to secrete extracellular matrix to connect to their neighbors, and the matrix disappeared when cells passed to the stationary growth phase. Cationized-ferritin staining resulted in a dense assembly of bulbs, protuberance-like structures on the growing bacterial cell surfaces. The cell-associated proteins derived by sonication contained predominated xylanase with relatively low carboxymethyl-cellulase (CMCase activities, suggesting that the xylanolytic-cellulolytic enzyme system occurred as a cell-associated enzyme. By means of gel-filtration chromatography, a high molecular mass protein with the estimated size of 2000 kDa was retrieved from the cell-associated enzymes, and it appeared as a single protein band on non-denaturing gel. However, more bands were obtained after the protein was boiled with sodium dodecyl sulfate and β-mercaptoethanol – which contained 4 xylanases and one CMCase – suggesting that these proteins were organized as a multienzyme complex (MEC in natural form. Additionally, the predominated xylanolytic MEC preferred binding to cellulose rather than xylan.

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

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    RODICA CHEREJI

    2013-07-01

    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.

  6. 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.

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

    International Nuclear Information System (INIS)

    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)

  8. Production, purification, and characterization of a cellulase-free thermostable endo-xylanase from Thermoanaerobacterium thermosaccharolyticum DSM 571.

    Science.gov (United States)

    Li, Xun; Shi, Hao; Ding, Huaihai; Zhang, Yu; Wang, Fei

    2014-12-01

    This is the first report describing the cloning, expression, and characterization of a putative thermostable, cellulase-free xylanase (XYN) from the thermophilic bacterium Thermoanaerobacterium thermosaccharolyticum DSM 571. The temperature and pH values for optimal enzyme activity of XYN were found to be 65 °C and pH 6.5, respectively. The XYN activity was apparently enhanced by Co(2+), Mn(2+), and Tween 60 and significantly inactivated by Al(3+), Cu(2+), Zn(2+), and SDS. The K m and V max values of XYN for the hydrolysis of beechwood xylan were 2.1 mg/ml and 222.1 U/mg, respectively. The k cat values of XYN for beechwood xylan at the optimal temperature and pH values were 481.4 s(-1). XYN represents an attractive candidate for use in the large-scale production of xylooligosaccharides (XOs) from forest residues because it is an endo-xylanase capable of degrading xylan. PMID:25261357

  9. Yeasts in mixed deciduous forest areas of Phujong Nayoy National Park and their ability to produce xylanase and carboxymethyl cellulase

    Directory of Open Access Journals (Sweden)

    Jantaporn Thongekkaew,

    2012-04-01

    Full Text Available A total of 61 yeast strains were obtained from 132 samples collected from various sources such as soil, mushroom,flowers, fruits, tree barks and insect frass in the mixed deciduous forest areas of Phujong Nayoy National Park, Thailand.Based on D1/D2 region at the 5 end of the large subunit ribosomal RNA gene (rRNA gene region D1/D2 analysis, 39 strainswere identified as ascomycetous yeasts and distributed to 7 genera i.e. Blastobotrys, Candida, Debaryomyces, Dipodascus,Kodamaea, Pichia and Torulaspora. Twenty strains were identified as basidiomycetous yeasts which belonged to the generaAsterotremella, Cryptococcus, Sporidiobolus and Trichosporon. Another two strains of yeast-like fungi were belonged togenus Aureobasidium. The predominant genus was Candida with a 31.14% contribution. For testing of xylanase and carboxymethylcellulase production of the 61 strains of yeasts and yeast-like fungi, Candida glabrata and Aureobasidiumpullulans showed xylanase activity of 0.91 and 0.52 UmL-1, respectively, and carboxymethyl cellulase activity of 0.38 and0.44 UmL-1, respectively.

  10. Indirect enzyme-antibody sandwich enzyme-linked immunosorbent assay for quantification of TAXI and XIP type xylanase inhibitors in wheat and other cereals.

    Science.gov (United States)

    Beaugrand, Johnny; Gebruers, Kurt; Ververken, Cedric; Fierens, Ellen; Dornez, Emmie; Goddeeris, Bruno M; Delcour, Jan A; Courtin, Christophe M

    2007-09-19

    To quantify Triticum aestivum xylanase inhibitor (TAXI) and xylanase inhibiting protein (XIP) type proteins in cereals in general and wheat ( T. aestivum) in particular, a robust enzyme-linked immunosorbent assay (ELISA) using an uncommon enzyme-antibody sandwich format was developed. Bacillus subtilis glycoside hydrolase family (GH) 11 and Aspergillus oryzae GH 10 xylanases were selected for coating ELISA plate wells to capture TAXI and XIP, respectively, prior to probing with antibodies. The detection threshold of the developed ELISA was much lower than that of the currently used xylanase inhibitor assay and the recently described Western blot approach. Because of its broad dynamic range (TAXI, 30-600 ng/mL, and XIP, 3-60 ng/mL), one proper standard extract dilution can be used for analyzing different wheat varieties, whereas for the currently used colorimetric assay, often different dilutions need to be analyzed. The TAXI ELISA for wheat was successfully adapted for barley ( Hordeum vulgare) and could also be used for other cereals. PMID:17715986

  11. Production of thermo-alkali-stable xylanase by a novel polyextremophilic Bacillus halodurans TSEV1 in cane molasses medium and its applicability in making whole wheat bread.

    Science.gov (United States)

    Kumar, Vikash; Satyanarayana, T

    2014-06-01

    A high titre of thermo-alkali-stable xylanase was attained in cane molasses medium. When the culture variables for endoxylanase production were optimized [cane molasses 7 %, soluble alkaline extract of wheat bran (SAE-WB) 37 % and ammonium chloride 0.30 %], a 4.5-fold enhancement in xylanase production (69 U ml(-1)) was achieved as compared to that in the unoptimized medium (15 U ml(-1)). The enzyme titre attained in shake flasks could be sustained in a 7-l laboratory bioreactor. An activity band corresponding to 40 kDa was visualized on SDS-PAGE zymogram analysis. The enzyme has broad range of pH and temperature for activity with optima at 9.0 and 80 °C, and stable between pH 4.0 and 11.0 with 85 % retention of activity. It has T 1/2 of 40 and 15 min at 70 and 80 °C. The enzyme is halotolerant since it displays activity in the presence of salt up to 15 %, and remains 100 % active in the absence of salt. The supplementation of whole wheat dough with xylanase improves antistaling property, reducing sugar content, bread volume with prebiotic xylooligosaccharides in bread. This is the first report on xylanase production in cane molasses medium with SAE-WB as the inducer and its applicability in whole wheat bread making that improves human health. PMID:24297158

  12. Preparation, Purification, and Secondary Structure Determination of Bacillus Circulans Xylanase. A Molecular Laboratory Incorporating Aspects of Molecular Biology, Biochemistry, and Biophysical Chemistry

    Science.gov (United States)

    Russo, Sal; Gentile, Lisa

    2006-01-01

    A project module designed for biochemistry or cellular and molecular biology student which involves determining the secondary structure of Bacillus circulans xylanase (BCX) by circular dichroism (CD) spectroscopy under conditions that compromise its stabilizing intramolecular forces is described. The lab model enhanced students knowledge of the…

  13. The effective expression of xylanase gene in Candida utilis by 18S rDNA targeted homologous recombination in pGLR9K.

    Science.gov (United States)

    Wei, Wang; Hong-Lan, Yang; HuiFang, Bao; Daoyuan, Zhang; Qi-mu-ge, Shan; Woof, Andrew J

    2010-07-01

    In order to test whether 18S rDNA can influence positively xylanase gene effective expression in the yeast of Candida utilis, a targeting vector pGLR9K-XA was constructed by adding an interested gene xynA from Streptomyces olivaceoviridis into the vector pGLR9K which is constructed by ourselves. pGLR9K contains the 18S rDNA, GAP promoter and CYH resistance gene sequence, all of which is from C. utilis. Then the vector pGLR9K-XA was transformed into C. utilis. To test the vector and transformed system, PCR, Southern blot and DNS methods were used. The results showed that xylanase gene can be detected in the chromosome DNA of recombinant C. utilis and the enzyme activity of xylanase is up to 60 IU ml(-1) in the study. It is suggested that this system can be used to express exogenous genes in C. utilis as a bioreactors. This is the first report that xylanase gene was expressed in C. utilis. PMID:19731075

  14. Biochemical and Thermodynamic Characterization of a Novel, Low Molecular Weight Xylanase from Bacillus Methylotrophicus CSB40 Isolated from Traditional Korean Food.

    Science.gov (United States)

    Panthi, Sandesh; Choi, Yoon Seok; Choi, Yun Hee; Kim, MiRi; Yoo, Jin Cheol

    2016-04-01

    A low molecular weight xylanase from Bacillus strain CSB40, isolated from traditional Korean food and produced in beechwood xylan, was biochemically and thermodynamically characterized. It was purified 8.12-fold with a 15.88 % yield using DEAE sepharose fast flow, and it was determined to have a mass of ∼27 kDa via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and xylan zymography. The purified xylanase was optimally active at 50 °C and pH 6 and stable over a wide range of pH (4.5-12.5). The N-terminal amino acid sequence of xylanase was GIQQGDDGKL. The activation energy for beechwood xylan hydrolysis was 29.39 kJmol(-1) with k cat value of 927.582 × 10(2) s(-1). K m and V max were 0.080 mg/ml and 794.63 mmol min(-1) mg(-1). The analysis of other thermodynamic parameters like ∆H, ∆G, ∆S, Q10, ∆GE-S, and ∆GE-T also supported the spontaneous formation of products, greater hydrolytic efficiency, and feasibility of enzymatic reaction, which also ratifies the novelty of this xylanase. The enzyme was strongly activated by Zn(2+) and inhibited by Cu(2+). The principal hydrolyzed end-products of this xylanase are xylobiose, xylotriose, and xylotetrose, which can be used in the pharmaceutical industry and as prebiotic in food. PMID:26780766

  15. 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 .

  16. Modification in the properties of paper by using cellulase-free xylanase produced from alkalophilic Cellulosimicrobium cellulans CKMX1 in biobleaching of wheat straw pulp.

    Science.gov (United States)

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

    2015-09-01

    Alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost is an actinomycete that produces industrially important and environmentally safer thermostable cellulase-free xylanase, which is used in the pulp and paper industry as an alternative to the use of toxic chlorinated compounds. Strain CKMX1 was previously characterized by metabolic fingerprinting, whole-cell fatty acids methyl ester analysis, and 16S rDNA and was found to be C. cellulans CKMX1. Crude enzyme (1027.65 U/g DBP) produced by C. cellulans CKMX1, having pH and temperature optima of 8.0 and 60 °C, respectively, in solid state fermentation of apple pomace, was used in the production of bleached wheat straw pulp. Pretreatment with xylanase at a dose of 5 U/g after pulping decreased pulp kappa points by 1.4 as compared with the control. Prebleaching with a xylanase dose of 5 U/g pulp reduced the chlorine charge by 12.5%, increased the final brightness points by approximately 1.42% ISO, and improved the pulp strength properties. Xylanase could be substituted for alkali extraction in C-Ep-D sequence and used for treating chemically bleached pulp, resulting in bleached pulp with higher strength properties. Modification of bleached pulp with 5 U of enzyme/g increased pulp whiteness and breaking length by 1.03% and 60 m, respectively; decreased tear factor of pulp by 7.29%; increased bulk weight by 3.99%, as compared with the original pulp. Reducing sugars and UV-absorbing lignin-derived compound values were considerably higher in xylanase-treated samples. Cellulosimicrobium cellulans CKMX1 has a potential application in the pulp and paper industries. PMID:26220821

  17. Xylanase production by Streptomyces viridosporus T7A in submerged and solid-state fermentation using agro-industrial residues

    Directory of Open Access Journals (Sweden)

    Luiz Romulo Alberton

    2009-11-01

    Full Text Available The study of xylanase production was conducted by Streptomyces viridosporus T7A in submerged (SmF and solid-state fermentation (SSF, using agro-industrial residues and sub-products. Napier grass, sugarcane bagasse and soybean bran were used as carbon source, substrate/support, and nitrogen source, respectively. In SmF, Napier grass (1% v/w supplemented with soybean bran, hydroxyethylcellulose and B complex vitamins were used. Soybean bran (1.5 % w/v, B complex vitamins (0.1%, and hydroxyethilcellulose (0.15% led to an increase in xylanase production (23.41 U/mL. In SSF, the effects of the following parameters were studied: substrate composition (sugarcane bagasse, Napier grass and soybean bran, initial moisture, and inoculum rate. In SSF, the highest xylanase activity (423.9 U/g was reached with: 70 % sugarcane bagasse, 20% Napier grass and 10% soybean meal, 90% of moisture, and 10(7/g substrate.A produção de xilanase por Streptomyces viridosporus T7A foi realizada em fermentação submersa (FSm e fermentação no estado sólido (FES utilizando resíduos e sub-produtos agroindustriais. Capim Napier, bagaço de cana e farelo de soja foram empregados como fonte de carbono, suporte/substrato e fonte nitrogênio, respectivamente. Em FSm, o capim Napier (1 % p/v foi suplementado com farelo de soja (1,5 % p/v, hidroxietilcelulose (0,15 % e vitaminas do complexo B (1,5 % sendo que a produção de xilanase atingiu 23.41 U/mL. Em FES, o efeito dos seguintes parâmetros foi estudado: composição do substrato (bagaço de cana, Capim Napier e farelo de soja, umidade inicial, aeração e taxa de inoculação. A mais elevada produção de xilanase (423,9 U/g foi atingida com 70% bagaço de cana, 20% de capim Napier e 10 % farelo de soja, 90 % de umidade inicial e 10(7 células/g substrato.

  18. SCREENING OF HARDWOOD AND SOFTWOOD SPECIES AS BEST SUBSTRATE FOR CELLULASE AND XYLANASE PRODUCTION USING CONSORTIUM OF POTENTIAL ISOLATES BACILLUS COAGULANS B30 + PAENIBACILLUS MUCILAGINOUS B5 + BACILLUS SP. UNDER SSF

    OpenAIRE

    Richa Kaushal*, Nivedita Sharma and Divya Tandon

    2014-01-01

    The lignocellulosic biomass is known to be an excellent carbon source for microbial enzyme production. In this paper, the cellulase and xylanase production from lignocellulosic materials using consortia of potential hydrolytic bacteria i.e. Bacillus coagulans B30 + Paenibacillus mucilaginous B5 + Bacillus sp. B21 isolated from forest soil under solid state fermentation (SSF) was investigated. The maximum cellulase activity of 97.84 U/g and xylanase activity of 67.06 U/g were obtained with wat...

  19. Effect of temperature on the production of cellulases, xylanases and lytic enzymes by selected Trichoderma reesei mutants

    Directory of Open Access Journals (Sweden)

    Piotr Janas

    2014-08-01

    Full Text Available The effect of temperature in the rangę of 26-38°C on the production of cellulases, xylanases and lytic enzymes by four mutant strains of Trichoderma reesei was analysed. On the basis of these investigations three thermosensitive strains (M-7. RUT C 30 and VTT-D-78085 which showed reduced excretion of the above mentioned enzymes as well as protein and a thermoresistant mutant (VTT-D-79I24 which grew within a temperature range of 26-34°C were characterized. Higher temperature caused an increase in the level of xylanolytic enzymes produced by the four mutants. In addition. it effected the complex composition of cellulolytic enzymes secreted by VTT-D-79l 24 (i.c. increased and reduced excertion of (β-glucosidase and β-1,4-endoglucanase respectively.

  20. Improved Enzyme Catalytic Characteristics upon Glutaraldehyde Cross-Linking of Alginate Entrapped Xylanase Isolated from Aspergillus flavus MTCC 9390

    Science.gov (United States)

    Bhushan, Bharat; Pal, Ajay; Jain, Veena

    2015-01-01

    Purified fungal xylanase was entrapped in alginate beads. Its further cross-linking using glutaraldehyde resulted in large enzyme aggregates which may function as both a catalyst and a support material for numerous substrate molecules. Enzyme cross-linking presented a negative impact on enzyme leaching during repeated washings and recovery of enzyme activity was substantial after twelve cycles of usage. The entrapment followed by cross-linking doubled the total bound activity and also greatly improved the enzyme stability at extreme chemical environment. The wide pH stability, better thermo- and storage stability, lowered Km value, and protection from some metal ions are salient achievements of present immobilization. The study shows the efficacy, durability, and sustainability of immobilized catalytic system which could be efficiently used for various juice processing operations. PMID:26347814

  1. Xylanase production with xylan rich lignocellulosic wastes by a local soil isolate of Trichoderma viride Produção de xilanase com resíduos lignocelulósicos ricos em xilana por uma cepa local de Trichoderma viride isolada de solo

    OpenAIRE

    Meenakshi Goyal; Kalra, K. L.; V.K. Sareen; G. Soni

    2008-01-01

    In the present study, cultural and nutritional conditions for enhanced production of xylanase by a local soil isolate of Trichoderma viride, using various lignocellulosic substrates in submerged culture fermentation have been optimized. Of the lignocellulosics used, maize straw was the best inducer followed by jowar straw for xylanase production. The highest activity achieved was between 14 to 17 days of fermentation. A continuous increase in xylanase production was observed with increasing l...

  2. Expression, crystallization and preliminary X-ray diffraction studies of thermostable β-1,3-xylanase from Thermotoga neapolitana strain DSM 4359

    International Nuclear Information System (INIS)

    Highly thermostable β-1,3-xylanase from T. neapolitana strain DSM 4359 was prepared and crystallized. X-ray diffraction data were collected to a resolution of 1.82 Å. Crystals of β-1,3-xylanase (1,3-β-d-xylan xylanohydrolase; EC 3.2.1.32) from Thermotoga neapolitana strain DSM 4359 with maximum dimensions of 0.2 × 0.1 × 0.02 mm were grown using the sitting-drop vapour-diffusion method at 293 K over 24 h. The crystals diffracted to a resolution of 1.82 Å, allowing structure determination. The crystals belonged to space group P21, with unit-cell parameters a = 39.061, b = 75.828, c = 52.140 Å; each asymmetric unit cell contained a single molecule

  3. Effect of wheat inclusion and xylanase supplementation of the diet on intestinal enzyme activity, nutrient retention and performance in laying hen from 25 to 47 wks of age

    OpenAIRE

    Mirzaie, S.; Zaghari, M.; Aminzadeh, S.; M Shivazad; Perez Serrano, Martina; Gonzalez Mateos, Gonzalo

    2011-01-01

    A trial was conducted to examine the effects of increasing levels of wheat in the diet and xylanase (ES) supplementation on nitrogen and ether extract retention, pH of the GIT, productive performance from 25 to 47 wks of age, and enzyme activity at the small intestine level. The basal diets (from 25 to 33 wks and from 33 to 47 wks) consisted of soybean meal and corn, and the wheat was introduced in the experimental diets at expenses of corn, primarily.

  4. Control on the Wheat Non-Starch Polysaccharides (NSP’ Anti-Nutritional Effect on Intestinal Wall, by Introducing Xylanase in Broiler Feed

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    Gabi Dumitrescu

    2011-10-01

    Full Text Available The experiment was performed in order to determine the protocol of xylanase utilization to fight against the antinutritional effect exerted by wheat non-starch polysaccharides (NSP on intestinal wall in broilers. The experimental works were carried out on broilers, the hybrid Ross 308. We formed four experimental groups, as follows: the experimental group LE1, fed on forage without wheat in its structure, the experimental group LE2, fed on combined forage including wheat in a proportion of 40%, the experimental group LE3 including wheat in a proportion of 40% and xylanase, in an amount of 25 g / to, and the experimental group LE4, including wheat in a proportion of 40% and xylanase, in an amount of 100 g. At 3 and 6 weeks, successive to chicken killing, we sampled the intestinal wall and determined the main changes occurred. The histo-morpho-metric analysis of the four experimental groups led to the conclusions that: wheat administration in a proportion of 40% in the individuals in LE2 determines the development of vilositary muscle elements, decreased of intestinal villosities height, leucocitary migration, and also vascular ectasies and reduced hemorrhagic areas; xylanase addition in the wheat-based feed may be associated with the increase of intestinal villosities height, and especially at jejuna level the villosities seem slightly branched, a slight hypertrophy of the epithelial cells compared with the individuals in LE2, the increase of goblet cells frequency and hypertrophy of the capillary network. These microscopic aspects come together with more intens digestion and absorption processes, and especially in the experimental group 4.

  5. Control on the Wheat Non-Starch Polysaccharides (NSP)’ Anti-Nutritional Effect on Intestinal Wall, by Introducing Xylanase in Broiler Feed

    OpenAIRE

    Gabi Dumitrescu; Lavinia Ştef; Dan Drinceanu; Calin Julean; Ducu Stef; Liliana Petculescu Ciochina; Cosmin Pandur

    2011-01-01

    The experiment was performed in order to determine the protocol of xylanase utilization to fight against the antinutritional effect exerted by wheat non-starch polysaccharides (NSP) on intestinal wall in broilers. The experimental works were carried out on broilers, the hybrid Ross 308. We formed four experimental groups, as follows: the experimental group LE1, fed on forage without wheat in its structure, the experimental group LE2, fed on combined forage including wheat in a proportion of 4...

  6. 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)

    The use of congress grass (Parthenium sp.) and water hyacinth (Eichhornia crassipes) as low cost raw materials for xylanase production from mutant Penicillium oxalicum SAUE-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

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

    Science.gov (United States)

    Rasala, Beth A; Lee, Philip A; Shen, Zhouxin; Briggs, Steven P; Mendez, Michael; Mayfield, Stephen P

    2012-01-01

    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. PMID:22937037

  8. 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.

  9. 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)

  10. GH10 XynA is the main xylanase identified in the crude enzymatic extract of Paenibacillus sp. A59 when grown on xylan or lignocellulosic biomass.

    Science.gov (United States)

    Ghio, Silvina; Insani, Ester M; Piccinni, Florencia E; Talia, Paola M; Grasso, Daniel H; Campos, Eleonora

    2016-01-01

    A novel bacterial isolate with polysaccharides degrading activity was identified as Paenibacillus sp., and named Paenibacillus sp. A59. Even though it is a strict mesophile, optimal xylanase activity of the crude enzymatic extract was achieved between 50°C and 70°C and more than 60% of the activity was retained after incubation for 48h at 50°C, indicating thermotolerance of the enzymes involved. The extract was also active on pre-treated sugarcane residue (SCR) and wheat straw, releasing xylobiose and xylose as the main products, therefore confirming its predominantly xylanolytic activity. By zymograms and mass spectrometry of crude enzymatic extracts of xylan or SCR cultures, a 32kDa GH10 beta- 1,4- endoxylanase with xylanase and no CMCase activity was identified. We named this enzyme XynA and it was the only xylanase identified under both conditions assayed, suggesting that it is a good candidate for recombinant expression and evaluation in hemicelluloses deconstruction applications. Also, a protein with two S-layer homology domains (SLH) and a large uncharacterized C-terminal domain as well as an ABC substrate binding protein were identified in crude extracts of SCR cultures. We propose that Paenibacillus sp. A59 uses a system similar to anaerobic and other Gram positive bacteria, with SLH-domain proteins anchoring polysaccharide-degrading enzymes close to the membrane and the substrate binding protein assisting translocation of simple sugars to the cell interior. PMID:27242139

  11. C-Terminal proline-rich sequence broadens the optimal temperature and pH ranges of recombinant xylanase from Geobacillus thermodenitrificans C5.

    Science.gov (United States)

    Irfan, Muhammad; Guler, Halil Ibrahim; Ozer, Aysegul; Sapmaz, Merve Tuncel; Belduz, Ali Osman; Hasan, Fariha; Shah, Aamer Ali

    2016-09-01

    Efficient utilization of hemicellulose entails high catalytic capacity containing xylanases. In this study, proline rich sequence was fused together with a C-terminal of xylanase gene from Geobacillus thermodenitrificans C5 and designated as GthC5ProXyl. Both GthC5Xyl and GthC5ProXyl were expressed in Escherichia coli BL21 host in order to determine effect of this modification. The C-terminal oligopeptide had noteworthy effects and instantaneously extended the optimal temperature and pH ranges and progressed the specific activity of GthC5Xyl. Compared with GthC5Xyl, GthC5ProXyl revealed improved specific activity, a higher temperature (70°C versus 60°C) and pH (8 versus 6) optimum, with broad ranges of temperature and pH (60-80°C and 6.0-9.0 versus 40-60°C and 5.0-8.0, respectively). The modified enzyme retained more than 80% activity after incubating in xylan for 3h at 80°C as compared to wild -type with only 45% residual activity. Our study demonstrated that proper introduction of proline residues on C-terminal surface of xylanase family might be very effective in improvement of enzyme thermostability. Moreover, this study reveals an engineering strategy to improve the catalytic performance of enzymes. PMID:27444327

  12. Optimization of β-Glucosidase, β-Xylosidase and Xylanase Production by Colletotrichum graminicola under Solid-State Fermentation and Application in Raw Sugarcane Trash Saccharification

    Directory of Open Access Journals (Sweden)

    Ana L. R. L. Zimbardi

    2013-01-01

    Full Text Available Efficient, low-cost enzymatic hydrolysis of lignocellulosic residues is essential for cost-effective production of bioethanol. The production of β-glucosidase, β-xylosidase and xylanase by Colletotrichum graminicola was optimized using Response Surface Methodology (RSM. Maximal production occurred in wheat bran. Sugarcane trash, peanut hulls and corncob enhanced β-glucosidase, β-xylosidase and xylanase production, respectively. Maximal levels after optimization reached 159.3 ± 12.7 U g−1, 128.1 ± 6.4 U g−1 and 378.1 ± 23.3 U g−1, respectively, but the enzymes were produced simultaneously at good levels under culture conditions optimized for each one of them. Optima of pH and temperature were 5.0 and 65 °C for the three enzymes, which maintained full activity for 72 h at 50 °C and for 120 min at 60 °C (β-glucosidase or 65 °C (β-xylosidase and xylanase. Mixed with Trichoderma reesei cellulases, C. graminicola crude extract hydrolyzed raw sugarcane trash with glucose yield of 33.1% after 48 h, demonstrating good potential to compose efficient cocktails for lignocellulosic materials hydrolysis.

  13. Efficiency of new fungal cellulase systems in boosting enzymatic degradation of barley straw lignocellulose

    DEFF Research Database (Denmark)

    Rosgaard, L.; Pedersen, S.; Meyer, Anne Boye Strunge

    2006-01-01

    the catalytic glucose yields significantly as compared to those obtained with the benchmark Celluclast + Novozyme 188 blend. A comparison of glucose yields obtained on steam-pretreated barley straw and microcrystalline cellulose, Avicel, indicated that the yield improvements were mainly due to the presence......This study examined the cellulytic effects on steam-pretreated barley straw of cellulose-degrading enzyme systems from the five thermophilic fungi Chaetomium thermophilum, Thielavia terrestris, Thermoascus aurantiacus, Corynascus thermophilus, and Myceliophthora thermophila and from the mesophile...

  14. Cloning and expression of the xynA1 gene encoding a xylanase of the GH10 group in Caulobacter crescentus.

    Science.gov (United States)

    Graciano, Luciana; Corrêa, Juliana Moço; Vieira, Fabíola Giovanna Nesello; Bosetto, Adilson; Loth, Eduardo Alexandre; Kadowaki, Marina Kimiko; Gandra, Rinaldo Ferreira; Simão, Rita de Cássia Garcia

    2015-04-01

    Caulobacter crescentus (NA1000 strain) are aquatic bacteria that can live in environments of low nutritional quality and present numerous genes that encode enzymes involved in plant cell wall deconstruction, including five genes for β-xylosidases (xynB1-xynB5) and three genes for xylanases (xynA1-xynA3). The overall activity of xylanases in the presence of different agro-industrial residues was evaluated, and it was found that the residues from the processing of corn were the most efficient in inducing bacterial xylanases. The xynA1 gene (CCNA_02894) encoding a predicted xylanase of group 10 of glyco-hydrolases (GH10) that was efficiently overexpressed in Escherichia coli LMG194 using 0.02 % arabinose, after cloning into the vector pJet1.2blunt and subcloning into the expression vector pBAD/gIII, provided a fusion protein that contained carboxy-terminal His-tags, named XynA1. The characterization of pure XynA1 showed an enzymatic activity of 18.26 U mL(-1) and a specific activity of 2.22 U mg-(1) in the presence of xylan from beechwood as a substrate. XynA1 activity was inhibited by EDTA and metal ions such as Cu(2+) and Mg(2+). By contrast, β-mercaptoethanol, dithiothreitol (DTT), and Ca(2+) induced recombinant enzyme activity. Kinetic data for XynA1 revealed K M and V max values of 3.77 mg mL-(1) and 10.20 μM min-(1), respectively. Finally, the enzyme presented an optimum pH of 6 and an optimum temperature of 50 °C. In addition, 80 % of the activity of XynA1 was maintained at 50 °C for 4 h of incubation, suggesting a thermal stability for the biotechnological processes. This work is the first study concerning the cloning, overexpression, and enzymatic characterization of C. crescentus xylanase. PMID:25791579

  15. Partial purification and properties of cellulase-free alkaline xylanase produced by Rhizopus stolonifer in solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Antonio José Goulart

    2005-05-01

    Full Text Available Rhizopus stolonifer was cultivated in wheat bran to produce a cellulase-free alkaline xylanase. The purified enzyme obtained after molecular exclusion chromatography in Sephacryl S-200 HR showed optimum temperature as 45º C and hydrolysis pHs optima as pH 6.0 and 9.0. Xylanase presented higher Vmax at pH 9.0 (0.87 µmol/mg protein than at pH 6.0 and minor Km at pH 6.0 (7.42 mg/mL than at pH 9.0.Rhizopus stolonifer foi cultivado em meio de farelo de trigo para produzir uma xilanase alcalina celulase-free. Uma amostra parcialmente purificada desta enzima foi obtida após cromatografia de exclusão molecular em Sephacryl S-200 HR. A temperatura ótima de hidrólise determinada (45º C está dentro do intervalo citado na literatura (45º C a 60º C para xilanases microbianas. Quanto ao pH ótimo, a amostra obtida apresentou atividades máximas em pH 6,0 e 9,0. Estes dados diferem da literatura, uma vez que o pH ótimo citado para a maioria das xilanases estudadas varia entre 4,0 e 5,5. De acordo com os estudos cinéticos realizados, a xilanase apresentou maior Vmax em pH 9,0 (0,87 µmol/mg proteína e menor Km em pH 6,0 (7,42 mg/mL. Os dois pHs ótimos determinados podem indicar a presença de isoformas desta enzima. Estes dados são interessantes pelo fato de que enzimas xilanolíticas alcalinas celulase-free podem ser utilizadas para o biobranqueamento da polpa na indústria de papel.

  16. Novel Cold-adaptive Penicillium Strain FS010 Secreting Thermo-labile Xylanase Isolated from Yellow Sea

    Institute of Scientific and Technical Information of China (English)

    Yun-Hua HOU; Tian-Hong WANG; Hao LONG; Hui-Yuan ZHU

    2006-01-01

    A novel cold-adaptive xylanolytic Penicillium strain FS010 was isolated from Yellow Sea sediments. The marine fungus grew well from 4 to 20 ℃; a lower (0 ℃) or higher (37 ℃) temperature limits its growth. The strain was identified as Penicillium chrysogenum. Compared with mesophilic P. chrysogenum,the cold-adaptive fungus secreted the cold-active xylanase (XYL) showing high hydrolytic activities at low temperature (2-15 ℃) and high sensitivity to high temperature (>50 ℃). The XYL gene was isolated from the cold-adaptive P. chrysogenum FS010 and designated as xyl. The deduced amino acid sequence of the protein encoded by xyl showed high homology with the sequence of glycoside hydrolase family 10. The gene was subcloned into an expression vector pGEX-4T-1 and the encoded protein was overexpressed as a fusion protein with glutathione-S-transferase in Escherichia coli BL21. The expression product was purified and subjected to enzymatic characterization. The optimal temperature and pH for recombinantXYL was 25 ℃ and 5.5, respectively. Recombinant XYL showed nearly 80% of its maximal activity at 4 ℃ and was active in the pH range 3.0-9.5.

  17. GH11 xylanase from Emericella nidulans with low sensitivity to inhibition by ethanol and lignocellulose-derived phenolic compounds.

    Science.gov (United States)

    Silva, Caio de Oliveira Gorgulho; Aquino, Elaine Nascimento; Ricart, Carlos André Ornelas; Midorikawa, Gláucia Emy Okida; Miller, Robert Neil Gerard; Filho, Edivaldo Ximenes Ferreira

    2015-07-01

    An endo-β-1,4-xylanase (X22) was purified from crude extract of Emericella nidulans when cultivated on submerged fermentation using sugarcane bagasse as the carbon source. The purified protein was identified by mass spectrometry and was most active at pH and temperature intervals of 5.0-6.5 and 50-60°C, respectively. The enzyme showed half-lives of 40, 10 and 7 min at 28, 50 and 55°C, respectively, and pH 5.0. Apparent Km and Vmax values on soluble oat spelt xylan were 3.39 mg/mL and 230.8 IU/mg, respectively, while Kcat and Kcat/Km were 84.6 s(-1) and 25.0 s(-1) mg(-1) mL. Incubation with phenolic compounds showed that tannic acid and cinnamic acid had an inhibitory effect on X22 but no time-dependent deactivation. On the other hand, ferulic acid, 4-hydroxybenzoic acid, vanillin and p-coumaric acid did not show any inhibitory effect on X22 activity, although they changed X22 apparent kinetic parameters. Ethanol remarkably increased enzyme thermostability and apparent Vmax and Kcat values, even though the affinity and catalytic efficiency for xylan were lowered. PMID:26040589

  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. PMID:26672453

  19. Effect of fungal biofertilizer from Chaetomium globosum on growth and quality of strawberry%球毛壳菌生物菌肥对草莓生长和品质的影响

    Institute of Scientific and Technical Information of China (English)

    辛雅芬

    2013-01-01

    [Objective]The present study was conducted to investigate effect of different dosages of fungal biofertilizer from Chaetomium globosum on growth,yield and quality of strawberry to provide references for utilizing the fungal biofertilizer in the field in future.[Method]Zhangji strawberry and fungal biofertilizer from C.globosum were served as test materials.Percentage of colonization of endophytic fungus C.globosum ND35 was detected in strawberry.Effects of fungal biofertilizer with different dosages of C.globosum spores on biomassy,physiological quota of strawberry and production and quality of fruits were investigated and analyzed in the field and in laboratory.[Result]Endophytic fungus C.globosum ND35 was able to colonize within plant of strawberry.Colonization rate was positively related to spores dosages of fungal biofertilizer.Different treatment of fungal biofertilizer from C.globosum significantly influenced growth,yield and quality of strawberry.Applying fungal biofertilizer at 0.5 g/plant and 1.0 g/plant was able to promote growth and development of strawberry.[Conclusion]The 0.5-1.0 g/plant utilizing dosages of fungal biofertilizer from C.globosum was able to increase yield and improve quality of strawberry.%[目的]研究球毛壳菌生物菌肥不同用量对草莓植株生长、果实产量和品质的影响,为球毛壳菌生物菌肥在生产上的应用提供参考.[方法]在温室大棚中开展试验,按不同球毛壳菌生物菌肥施肥量(0.5、1.0、2.0 g/株)设3个处理,不施肥为对照处理,检测内生真菌球毛壳菌ND35在草莓上的定殖率;分析不同菌肥用量对草莓生物量和生理指标及果实产量和品质的影响.[结果]球毛壳菌ND35可以在草莓植株内定殖,其定殖率与菌肥用量正相关.不同处理的球毛壳菌生物菌肥对草莓植株生长、果实产量和品质具有显著影响,施用0.5 g/株和1.0 g/株的菌肥用量有利于草莓的生长和发育.[结论]0.5~1.0/株用量

  20. Effect of xylanases on ileal viscosity, intestinal fiber modification, and apparent ileal fiber and nutrient digestibility of rye and wheat in growing pigs

    DEFF Research Database (Denmark)

    Lærke, Helle Nygaard; Arent, Susan; Dalsgaard, Søren;

    2015-01-01

    Two experiments were performed to study the effect of xylanase on ileal extract viscosity, in vivo fiber solubilization and degradation, and apparent ileal digestibility (AID) of fiber constituents, OM, CP, starch, and crude fat in rye and wheat in ileal-cannulated pigs. In Exp. 1, coarse rye...... AID of arabinoxylan by 91% to 107% (P < 0.001) across cereal matrices. Enzyme addition did not affect AID of nutrients in any of the experiments except for a higher starch and crude fat digestibility of fine wheat with enzyme addition (P < 0.012) in Exp. 2. Collectively, the results suggest that...

  1. Effect of β-glucanase and xylanase supplementation of barley- and rye-based diets on caecal microbiota of broiler chickens

    DEFF Research Database (Denmark)

    Josefiak, Damian; Rutkowski, A; Kaczmarek, S;

    2010-01-01

    1. The aim was to investigate the effect of grain type (barley or rye) and exogenous enzymes (β-glucanase or xylanase) on the composition of chicken caecal microbiota as examined by classical culturing and molecular techniques (fluorescent in-situ hybridisation (FISH) and terminal...... T-RFLP profiles indicated that the caecal microbiota of birds receiving rye-based diets was more diverse than that of birds fed on barley-based diets. 5. Irrespective of the method applied, the results indicate that the cereal type as well as the exogenous enzyme supplementation influence the...... microbiota in broiler chicken caeca, and may have the effect of reducing potentially pathogenic Enterobacteriaceae populations....

  2. 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......, by use of four xylanases from two different glycoside hydrolase families, GH10 and GH11, along with protease and phytase. Wheat DDGS showed the highest degree of enzymatic degradation due to a lower degree of cell wall complexity compared with that of corn DDGS. For corn DDGS, the combination of...

  3. Methane production of two roughage and total mixed ration as influenced by cellulase and xylanase enzyme addition

    Directory of Open Access Journals (Sweden)

    Belete Shenkute Gemeda

    2015-02-01

    Full Text Available In recent decades supplementation of animal feeds with exogenous fibrolytic enzymes has substantially improved digestibility and animal performance. However, information related to associated methane production is limited and inconsistent. This study evaluated the effect of cellulase and xylanase enzymes on in vitro methane production of Eragrostis curvula hay, maize (Zea mays stover and a total mixed ration (TMR at seven levels of the two enzymes. Feed samples were incubated for 2, 12, 24 and 48 h in an in vitro batch culture with buffer and rumen fluid, and fibrolytic enzymes. Gas production was measured using a pressure transducer connected to a data tracker, while methane gas was analysed using a gas chromatograph which was calibrated with standard CH4 and CO2. Increases in the level of enzyme application resulted in increases in gas volume, total volatile fatty acid (VFA production, dry matter (DM disappearance and associated increases in methane production. The linear increase in percentage and volume of methane production in tandem with increases in level of enzyme application might be due to increased fermentation, and organic matter degradability that resulted in a shift in VFA production towards acetate. Considering the efficiency of DM and neutral detergent fiber degradation and production of associated VFA with levels of enzymes, the use of 1 mg g−1 DM of enzyme can be a good option for the feeds tested. However, they cannot decrease methane production. It will be very important to consider other hydrogen sinks that can capture directly extra H+ produced by the addition of enzyme so that their supplementation could be very efficient and environmentally sound.

  4. Evidence that the xylanase activity from Sulfolobus solfataricus Oalpha is encoded by the endoglucanase precursor gene (sso1354) and characterization of the associated cellulase activity.

    Science.gov (United States)

    Maurelli, Luisa; Giovane, Alfonso; Esposito, Alessandra; Moracci, Marco; Fiume, Immacolata; Rossi, Mosè; Morana, Alessandra

    2008-09-01

    Sulfolobus solfataricus strain Oalpha was previously isolated for its ability to grow on minimal medium supplemented with xylan as a carbon source. The strain exhibited thermostable xylanase activity but several attempts to identify the gene encoding for the activity failed. Further studies showed that the xylanase displayed activity on carboxymethylcellulose (CMC) and the new activity was characterized. It exhibited an optimal temperature and pH of 95 degrees C and 3.5, respectively, and a half-life of 53 min at 95 degrees C. The enzyme, which was demonstrated to be glycosylated, hydrolyzed CMC in an endo-manner releasing cellobiose and other cello-oligomers. Analysis of the tryptic fragments by tandem mass spectrometry led to identification of the endoglucanase precursor, encoded by the sso1354 gene, as the protein possessing dual activity. The efficiency of the SSO1354 protein in degrading cellulosic and hemicellulosic fractions contained in agronomic residues was tested at low pH and high temperature. Cellulose and xylan were degraded to glucose and xylose at 90 degrees C, pH 4 by an enzyme mix consisting of SSO1354 and additional glycosyl hydrolases from S. solfataricus Oalpha. Given its role in saccharification processes requiring high temperatures and acidic environments, SSO1354 represents an interesting candidate for the utilization of agro-industrial waste for fuel production. PMID:18568289

  5. BIO-CONVENTIONAL BLEACHING OF KRAFT-AQ PULP OF A. CADAMBA BY CRUDE XYLANASES FROM COPRINELLUS DISSEMINATUS MLK-03 AND EFFECT OF RESIDUAL ENZYME ON EFFLUENT LOAD

    Directory of Open Access Journals (Sweden)

    Mohan Lal

    2011-04-01

    Full Text Available A new thermo-alkali-tolerant crude xylanase from Coprinellus disseminatus decreased kappa number by 34.38% and improved brightness and viscosity by 1.6 and 6.47% respectively after XE1-stage during prebleaching of Anthocephalus cadamba kraft-AQ pulp. At 2.4% chlorine demand, crude xylanase in a XECEHH (X= enzymatic prebleaching stage, E= extraction stage, C= chlorination stage, H= hypochlorite stage bleaching sequence improved pulp brightness, tensile index, burst index, and double fold numbers by 3.66%, 4.78%, 6.38%, and 11.11%, respectively with a reduction in viscosity (10.59% and tear index (10.77% compared to the control. Combined bleach effluent of the XECEHH sequence mitigated adsorable organic halides (AOX by 21% and increased chemical oxygen demand (COD, bio-chemical oxygen demand (BOD, and colour by 67.18%, 84.78%, and 97.53%, respectively, compared to the control. Residual enzymes that entered during enzymatic prebleaching stage decreased AOX, COD, BOD, and colour of combined effluent of the XECEHH bleaching sequence progressively and on 6th day, and these were reduced by 23.78%, 0.04%, 15.00%, and 0.61%, respectively, compared to the control.

  6. 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)

    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

  7. 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 Au(3+) to Au(0). 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. PMID:26864877

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

    International Nuclear Information System (INIS)

    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 P3221, 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 P3221 and contain a dimer in the asymmetric unit. The crystals diffracted to beyond 2.0 Å resolution

  9. 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.

  10. High-level soluble expression of a thermostable xylanase from thermophilic fungus Thermomyces lanuginosus in Escherichia coli via fusion with OsmY protein.

    Science.gov (United States)

    Le, Yilin; Wang, Huilei

    2014-07-01

    A thermostable xylanase is encoded by xynA from fungus Thermomyces lanuginosus. The problem emerged from overexpression of xynA in Escherichia coli has been the formation of inclusion bodies. Here we describe the xynA was fused with the hyperosmotically inducible periplasmic protein of E. coli, OsmY. The fusion protein OsmY-xynA was expressed as almost all soluble form. The soluble expression level of fusion protein reached 98±6U/ml when cells containing pET-OsmY-xynA were expressed without IPTG induction at 37°C. The induction is probably due to auto-induction due to lactose in the medium (Studier (2005) [21]). The cells harboring pET-OsmY-xynA expressed an activity level about 24 times higher than that expressed from pET-20b-xynA. Xylanase activity was observed in the extracellular (36±1.3U/ml) and the periplasmic (42±4U/ml) when cells containing pET-OsmY-xynA were induced without IPTG addition. After the cold osmotic shock procedure followed by nickel affinity chromatography, the purified fusion protein showed a single band on SDS-PAGE gel with a molecular mass of 44kDa. The purified fusion enzyme exhibited the highest activity at 65°C and pH 6.0. PMID:24657705

  11. The effects of transportation stress on Japanese quail (Coturnix Coturnix japonica) fed corn-based diet in comparison with wheat-based diet supplemented with xylanase and phytase.

    Science.gov (United States)

    Mehraei Hamzekolaei, M H; Zamani Moghaddam, A K; Tohidifar, S S; Dehghani Samani, A; Heydari, A

    2016-08-01

    Harvesting, handling and transporting quails to the slaughterhouses, other farms and laboratories might covertly reduce their welfare. The aim of this study was to evaluate the effects of two major sources of energy in poultry nutrition on reducing transportation stress in Japanese quail (Coturnix Coturnix japonica). Male quails (n = 60) were divided into two groups. The first group was fed corn-based diet, and the second was fed wheat-based diet supplemented with xylanase and phytase. At the end of the experiment (day 35), quails were subjected to 80 km of transportation. Immediately on arrival and after 24 h, heterophil counts, lymphocyte counts and H:L ratios were measured. On arrival, H counts were lower, L counts were higher, and H:L ratios were lower for corn-fed group. After 24 h, wheat-fed group showed lower increment of H counts, greater increment of L counts and also decrement of H:L ratios rather than corn-fed group which showed increment of H:L ratios. However, these ratios were still lower in corn-fed group. Results indicate that corn-based diets can help Japanese quail to better resist transportation stress, although it seems that feeding wheat-based diets supplemented with xylanase and phytase could have positive effects for coping better with stress after journeys. PMID:26459218

  12. Evaluation of different lignocellulosic substrates for the production of cellulases and xylanases by the basidiomycete fungi Bjerkandera adusta and Pycnoporus sanguineus.

    Science.gov (United States)

    Quiroz-Castañeda, Rosa Estela; Pérez-Mejía, Nancy; Martínez-Anaya, Claudia; Acosta-Urdapilleta, Lourdes; Folch-Mallol, Jorge

    2011-06-01

    Agricultural waste products are potential resources for the production of a number of industrial compounds, including biofuels. Basidiomycete fungi display a battery of hydrolytic enzymes with prospective use in lignocellulosic biomass transformation, however little work has been done regarding the characterization of such activities. Growth in several lignocellulosic substrates (oak and cedar sawdust, rice husk, corn stubble, wheat straw and Jatropha seed husk) and the production of cellulases and xylanases by two basidiomycete fungi: Bjerkandera adusta and Pycnoporus sanguineus were analyzed. Growth for P. sanguineus was best in rice husk while corn stubble supported the highest growth rate for B. adusta. Among the substrates tested, cedar sawdust produced the highest cellulolytic activities in both fungal species, followed by oak sawdust and wheat straw. Xylanolytic activity was best in oak and cedar sawdust for both species. We found no correlation between growth and enzyme production. Zymogram analysis of xylanases and cellulases showed that growth in different substrates produced particular combinations of protein bands with hydrolytic activity. PMID:20963471

  13. Production of cellulase-free xylanase by Trichoderma reesei SAF3 Produção de xilanase livre de celulase por Trichoderma reesei SAF3

    Directory of Open Access Journals (Sweden)

    Sanjay Kar

    2006-12-01

    Full Text Available A xylanase producing fungi has been isolated from soil and identified as Trichoderma reesei SAF3. Maximum growth of the organism was found at 48 h under submerged condition in xylan containing enriched medium, whereas highest enzyme production (4.75U/mL was recorded at 72 h. No detectable cellulase activity was noted during whole cultivation period. The partially purified enzyme hydrolyzed xylan into xylopentose and xylose. All these properties of xylanase highlighten its promising uses in industrial scale.A partir de solo, isolou-se uma cepa de fungos produtos de xilanase, posteriormente identificado como Trichoderma reesei SAF3. O crescimento máximo do fungo foi obtido após 48h em condições submersas em meio de cultura contendo xilano, enquanto produção máxima de enzima (4,75U/mL ocorreu em 72h. Durante o período de cultivo, não foi detectada atividade celulásica. A enzima parcialmente purificada hidrolizou xilano a xilopentose e xilose. Essas propriedades da xilanase destacam seu uso promissor em escala industrial.

  14. Purification and characterization of a low molecular weight xylanase from solid-state cultures of Aspergillus fumigatus Fresenius Purificação e caracterização de uma xilanase de baixo peso molecular de culturas de estado sólido de Aspergillus fumigatus Fresenius

    OpenAIRE

    Claudio Henrique Cerri e Silva; Jurgen Puls; Marcelo Valle de Sousa; Edivaldo Ximenes Ferreira Filho

    1999-01-01

    A xylan-degrading enzyme (xylanase II) was purified to apparent homogeneity from solid-state cultures of Aspergillus fumigatus Fresenius. The molecular weight of xylanase II was found to be 19 and 8.5 kDa, as estimated by SDS-PAGE and gel filtration on FPLC, respectively. The purified enzyme was most active at 55 °C and pH 5.5. It was specific to xylan. The apparent Km and Vmax values on soluble and insoluble xylans from oat spelt and birchwood showed that xylanase II was most active on solub...

  15. SCREENING OF HARDWOOD AND SOFTWOOD SPECIES AS BEST SUBSTRATE FOR CELLULASE AND XYLANASE PRODUCTION USING CONSORTIUM OF POTENTIAL ISOLATES BACILLUS COAGULANS B30 + PAENIBACILLUS MUCILAGINOUS B5 + BACILLUS SP. UNDER SSF

    Directory of Open Access Journals (Sweden)

    Richa Kaushal*, Nivedita Sharma and Divya Tandon

    2014-05-01

    Full Text Available The lignocellulosic biomass is known to be an excellent carbon source for microbial enzyme production. In this paper, the cellulase and xylanase production from lignocellulosic materials using consortia of potential hydrolytic bacteria i.e. Bacillus coagulans B30 + Paenibacillus mucilaginous B5 + Bacillus sp. B21 isolated from forest soil under solid state fermentation (SSF was investigated. The maximum cellulase activity of 97.84 U/g and xylanase activity of 67.06 U/g were obtained with water containing pretreated Celtis australis. Additional nutrients of inorganic salts were added in the form of modified basal salt medium (BSM which resulted in increased enzyme production i.e. cellulase activity of 177.14 U/g and xylanase activity of 115.42 U/g using pretreated C. australis under solid-state fermentation (SSF. Results indicate the excellent scope of utilizing C. australis as solid substrate for commercial production of cellulase and xylanase employing consortium of Bacillus spp.

  16. ECF and TCF bleaching of Saccharum officinerum-CO89003 bagasse soda-AQ pulp with alkali-thermo-tolerant crude xylanase from Coprinellus disseminatus SW-1 NTCC1165

    Directory of Open Access Journals (Sweden)

    Swarnima Agnihotri

    2012-11-01

    Full Text Available An alkali-thermo-tolerant crude xylanase from Coprinellus disseminatus SW-1 NTCC1165 produced under solid-state fermentation conditions improves the brightness of sugarcane bagasse soda-AQ pulp by 7.3, 4.7, 6.1, and 8.2% in XODED, XOD(EOPDP, OX(EOPP, and XO(EOPP bleaching sequences, respectively, at an enzyme dose of 8IU/g, a reaction time of 120 min, a consistency of 10%, and a pH of 6.4 at 55 oC. An improvement in brightness by 2.1% for pulp bleached by XO(EOPP compared to OX(EOPP sequence validates that xylanase treatment is more effective for hydrolysing lignin-carbohydrates complexes before oxygen treatment. AOX after XODED and XOD(EOPDP sequences is reduced by 41.43 and 40%, respectively, compared to controls, but an increase in COD and color in studied bleaching sequences is attributable to the hydrolysis of hemicelluloses and the release of lignin-carbohydrates complexes after xylanase treatment. Xylanase treatment modifies fibre surface by introducing cracks, peelings, swelling, and external fibrillation, which facilitates faster penetration of bleach chemicals by disrupting physical barriers, as revealed by scanning electron microscopy.

  17. The degradation of arabinoxylan-rich cell walls in digesta obtained from piglets fed wheat-based diets varies depending on digesta collection site, type of cereal, and source of exogenous xylanase.

    Science.gov (United States)

    Pedersen, N R; Azem, E; Broz, J; Guggenbuhl, P; Le, D M; Fojan, P; Pettersson, D

    2012-12-01

    The objective of the present study was to compare the ability of experimental and commercial xylanases to degrade, in vitro, the arabinoxylan (AX) fraction in digesta from 28-d-old piglets fed a wheat (Triticum aestivum)-based diet (49% wheat). Pigs were euthanized at 1, 2, 3, or 4 h after feeding; stomach and ileum contents were isolated and frozen and later used for the in vitro studies. Xylan solubilization provided information regarding the ability of the enzymes to degrade AX during the harsh in vivo conditions prevailing in the gastrointestinal tract. The hydrolytic capacity of a commercial xylanase was compared with that of an experimental xylanase using stomach digesta (pH 1.8) obtained at 4 h after feeding. Relative to the control, both enzymes increased (P < 0.001) xylan solubilization 3-fold. In the ileal digesta (1 h), xylan solubilization was increased by 36% (P < 0.001). Inclusion of arabinofuranosidases (Ara f) with xylanases increased xylan solubilization in stomach samples (P = 0. 007 and P = 0. 030) but not in ileal samples (P = 0.873 and P = 0.997). Our results illustrate clearly the importance of using different conditions and substrates when enzyme performance is studied in vitro as a prescreening tool for setting up in vivo trials. PMID:23365312

  18. High-level recombinant protein production by the basidiomycetous yeast Pseudozyma antarctica under a xylose-inducible xylanase promoter.

    Science.gov (United States)

    Watanabe, Takashi; Morita, Tomotake; Koike, Hideaki; Yarimizu, Tohru; Shinozaki, Yukiko; Sameshima-Yamashita, Yuka; Yoshida, Shigenobu; Koitabashi, Motoo; Kitamoto, Hiroko

    2016-04-01

    Yeast host-vector systems are useful tools for the production of recombinant proteins. Here, we report the construction of a new high-level expression plasmid pPAX1-neo for the basidiomycetous yeast, Pseudozyma antarctica. pPAX1-neo harbours a xylose-inducible expression cassette under control of the xylanase promoter and terminator of P. antarctica T-34, a selection cassette of neomycin/G418 with an Escherichia coli neomycin resistance gene under control of the homocitrate synthase promoter of strain T-34, and an autonomously replicating sequence fragment of Ustilago maydis (UARS). Biodegradable plastic (BP)-degrading enzymes of P. antarctica JCM10317 (PaE) and Paraphoma-related fungal strain B47-9 (PCLE) were used as reporter proteins and inserted into pPAX1-neo, resulting in pPAX1-neo::PaCLE1 and pPAX1-neo::PCLE, respectively. Homologous and heterologous BP-degrading enzyme production of transformants of P. antarctica T-34 were detected on agar plates containing xylose and emulsified BP. Recombinant PaE were also produced by transformants of other Pseudozyma strains including Pseudozyma aphidis, Pseudozyma rugulosa, and Pseudozyma tsukubaensis. To improve the stability of transformed genes in cells, the UARS fragment was removed from linearized pPAX1-neo::PaCLE1 and integrated into the chromosome of the P. antarctica strain, GB-4(0), which was selected as a PaE producer in xylose media. Two transformants, GB-4(0)-X14 and X49, had an 11-fold higher activity compared with the wild type strain in xylose-containing liquid media. By xylose fed-batch cultivation using a 3-L jar fermentor, GB-4(0)-X14 produced 73.5 U mL(-1) of PaE, which is 13.4-fold higher than that of the wild type strain GB-4(0), which produced 5.5 U mL(-1) of PaE. PMID:26695155

  19. 小麦饲粮中添加木聚糖酶对肉鹅血糖和血清总蛋白水平的影响%Effect of Wheat Based Diet Supplemented with Xylanase on Blood Sugar and Total Protein in Serum of Geese

    Institute of Scientific and Technical Information of China (English)

    王佳丽; 史东辉; 杨桂芹

    2008-01-01

    [Objective] The effects of wheat based diet supplemented with xylarmse on blood sugar and total protein in serum of geese were studied. [Method] By using the randomized design of single factor, the 1-day-old healthy goslings were divided into 6 groups and fed with corn based diet, wheat based diet and wheat based diet supplemented with xylanase at different concentrations respectively, the contents of blood sugar and total protein in serum were determined. [Result] The wheat based diet supplemented with xylanase could increase the blood sugar and total protein in serum of geese and wheat based diet supplemented with 0.2% xylanase generated the best effect, which was higher than those of corn based diet group. As for the concentration of protein in senun, wheat based diet supplemented with O. 2% xylarmse was significantly different from corn based diet and wheat based diet. [Conclusion] The wheat based diet supplemented with xylanase could enhance geese production.

  20. Production and purification of an Endo–1,4-beta-Xylanase from Humicola grisea var. thermoidea by electroelution Produção e purificação de uma Endo–1,4-beta-Xylanase de Humicola grisea var. thermoidea por eletroeluição

    Directory of Open Access Journals (Sweden)

    Rubens Monti

    2003-06-01

    Full Text Available Humicola grisea var. thermoidea produces two forms of extracellular xylanase. The component form 1 was purified using the electroelution method, due to the very small production of this extracellular enzyme. The apparent molecular mass was 61.8 kDa by SDS-PAGE.Humicola grisea var. thermoidea produz duas formas de xilanase extracelular. A componente forma 1 foi purificada usando o método de eletroeluição devido à baixa produção desta enzima extracelular. A aparente massa molar foi determinada 61,8 kDa por SDS-PAGE.

  1. Production and purification of an Endo–1,4-beta-Xylanase from Humicola grisea var. thermoidea by electroelution Produção e purificação de uma Endo–1,4-beta-Xylanase de Humicola grisea var. thermoidea por eletroeluição

    OpenAIRE

    Rubens Monti; Leonardo Cardello; Marcos F. Custódio; Antonio J Goulart; Adriana H. Sayama; Jonas Contiero

    2003-01-01

    Humicola grisea var. thermoidea produces two forms of extracellular xylanase. The component form 1 was purified using the electroelution method, due to the very small production of this extracellular enzyme. The apparent molecular mass was 61.8 kDa by SDS-PAGE.Humicola grisea var. thermoidea produz duas formas de xilanase extracelular. A componente forma 1 foi purificada usando o método de eletroeluição devido à baixa produção desta enzima extracelular. A aparente massa molar foi determinada ...

  2. Production and characterization of an enzyme complex from a new strain of Clostridium thermocellum with emphasis on its xylanase activity Produção e caracterização de um complexo enzimático de uma nova linhagem de Clostridium thermocellum com enfase em sua atividade de xilanase

    OpenAIRE

    Werner Bessa Vieira; Leonora Rios de Souza Moreira; Amadeu Monteiro Neto; Edivaldo Ximenes Ferreira Filho

    2007-01-01

    A new bacterial strain (ISO II) was isolated from manure cow and identified as phylogenetically close to the thermophilic cellulolytic bacterium Clostridium thermocellum. The new strain produced extracellular xylanase, pectinase, mannanase and cellulase activities when grown in liquid culture medium containing banana stem as carbon source. The enzyme production profile after growth on banana stem showed that xylanase and cellulase activities were detected in different incubation periods. An e...

  3. 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.

  4. Calcium alginate matrix increases the stability and recycling capability of immobilized endo-β-1,4-xylanase from Geobacillus stearothermophilus KIBGE-IB29.

    Science.gov (United States)

    Bibi, Zainab; Qader, Shah Ali Ul; Aman, Afsheen

    2015-07-01

    Exploration of microbial pool from extremely diversified ecosystem is significantly important for various industrial applications. Bacterial communities from extreme habitats including volcanic vents, hot springs, and industrial sectors are eagerly explored for the isolation of thermophiles. Geobacillus stearothermophilus KIBGE-IB29, isolated from blast furnace site of a steel processing industry, is capable of producing thermostable endo-β-1,4-xylanase. In the current study, this enzyme was immobilized within calcium alginate beads using entrapment technique. Amalgamation of sodium alginate (40.0 gL(-1)) and calcium chloride (0.4 M) was used for the formation of immobilized beads. It was observed that temperature (50 °C) and pH (7.0) optima of immobilized enzyme remained same, but enzyme-substrate reaction time increased from 5.0 to 30.0 min as compared to free enzyme. Diffusion limit of high molecular weight xylan (corncob) caused a decline in V max of immobilized enzyme from 4773 to 203.7 U min(-1), whereas K m value increased from 0.5074 to 0.5722 mg ml(-1) with reference to free enzyme. Immobilized endo-β-1,4-xylanase showed its stability even at high temperatures as compared to free enzyme and retained 18 and 9 % residual activity at 70 and 80 °C, respectively. Immobilized enzyme also exhibited sufficient recycling efficiency up to five reaction cycles which indicated that this enzyme can be a plausible candidate in paper and pulp industry. PMID:26001519

  5. Studies on properties of the xylan‑binding domain and linker sequence of xylanase XynG1‑1 from Paenibacillus campinasensis G1‑1.

    Science.gov (United States)

    Liu, Yihan; Huang, Lin; Li, Weiguo; Guo, Wei; Zheng, Hongchen; Wang, Jianling; Lu, Fuping

    2015-12-01

    Xylanase XynG1-1 from Paenibacillus campinasensis G1-1 consists of a catalytic domain (CD), a family 6_36 carbohydrate-binding module which is a xylan-binding domain (XBD), and a linker sequence (LS)between them. The structure of XynG1-3 from Bacillus pumilus G1-3 consists only of a CD. To investigate the functions and properties of the XBD and LS of XynG1-1, two truncated forms (XynG1-1CDL, XynG1-1CD) and three fusion derivatives (XynG1-3CDL, XynG1-3CDX and XynG1-3CDLX) were constructed and biochemically characterized. The optimum conditions for the catalytic activity of mutants of XynG1-1 and XynG1-3 were 60 °C and pH 7.0, and 55 °C and pH 8.0, respectively, the same as for the corresponding wild-type enzymes. XynGs with an XBD were stable over a broad temperature (30-80 °C)and pH range (4.0-11.0), respectively, on incubation for 3 h. Kinetic parameters (Km, kcat, kcat/Km) of XynGs were determined with soluble birchwood xylan and insoluble oat spelt xylan as substrates. XynGs with the XBD showed better affinities toward, and more efficient catalysis of hydrolysis of the insoluble substrate. The XBD had positive effects on thermostability and pH stability and a crucial function in the ability of the enzyme to bind and hydrolyze insoluble substrate. The LS had little effect on the overall stability of the xylanase and no relationship with affinities for soluble and insoluble substrates or catalytic efficiency. PMID:26467249

  6. 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.

  7. Scientific Opinion on the safety and efficacy of AveMix® XG 10 (endo-1,4-beta-xylanase and endo-1,3(4-beta-glucanase as a feed additive for turkeys for fattening

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2013-04-01

    Full Text Available The additive AveMix® XG 10 is an enzyme preparation of endo-1,4-beta-xylanase (xylanase and endo-1,3(4-beta-glucanase (glucanase, produced by two strains of Trichoderma reesei. This product is currently authorised for use in chickens for fattening, laying hens, minor poultry species and weaned piglets as a zootechnical additive, under the functional group of digestibility enhancers. The applicant is now seeking an extension of the authorisation to turkeys for fattening at a recommended dose of 4 000 XU (xylanase units and 900 BGU (glucanase units per kg complete feed. The results obtained in a tolerance study in turkeys for fattening showed that the birds tolerated well a 100-fold overdose of the recommended dose. Therefore, the additive is safe for turkeys for fattening when used at the recommended dose. Three efficacy studies carried out in turkeys for fattening showed that the additive has the potential to be efficacious at the dose of 4 000 XU and 900 BGU/kg complete feed.

  8. A computational method for the systematic screening of reaction barriers in enzymes: Searching for Bacillus circulans xylanase mutants with greater activity towards a synthetic substrate

    CERN Document Server

    Hediger, Martin R; De Vico, Luca; Jensen, Jan H

    2013-01-01

    We present a semi-empirical (PM6-based) computational method for systematically estimating the effect of all possible single mutants, within a certain radius of the active site, on the barrier height of an enzymatic reaction. The intent of this method is not a quantitative prediction of the barrier heights, but rather to identify promising mutants for further computational or experimental study. The method is applied to identify promising single and double mutants of Bacillus circulans xylanase (BCX) with increased hydrolytic activity for the artificial substrate ortho-nitrophenyl \\beta-xylobioside (ONPX$_2$). The estimated reaction barrier for wild-type (WT) BCX is 18.5 kcal/mol, which is in good agreement with the experimental activation free energy value of 17.0 kcal/mol extracted from the observed k$_\\text{cat}$ using transition state theory (Joshi et al., Biochemistry 2001, 40, 10115). The PM6 reaction profiles for eight single point mutations are recomputed using FMO-MP2/PCM/6-31G(d) single points. PM6 ...

  9. Evaluation of high dietary inclusion of distillers dried grains with solubles and supplementation of protease and xylanase in the diets of broiler chickens under necrotic enteritis challenge.

    Science.gov (United States)

    Barekatain, M R; Antipatis, C; Rodgers, N; Walkden-Brown, S W; Iji, P A; Choct, M

    2013-06-01

    A 2 × 2 × 2 factorial experiment was conducted to investigate the effect of a high level of sorghum distillers dried grains with solubles (DDGS; 20%), with or without a combination of protease and xylanase in broiler chickens, under a necrotic enteritis disease challenge. A total of 576 male broiler chicks were randomly assigned to 8 experimental treatments, each replicated 6 times, with 12 birds per replicate for 35 d. Oral inoculation of the challenged group with Eimeria spp. occurred on d 9, followed by 3 consecutive inoculations of Clostridium perfringens from d 14 through 16. The disease challenge and DDGS inclusion significantly (P perfringens in the ceca at d 17. Inoculation of birds with C. perfringens resulted in higher (P perfringens in both ileal and cecal contents. The necrotic enteritis-related lesions (d 17) were more severe (P chickens may increase susceptibility to necrotic enteritis. Supplementation of enzymes did not reveal significant mitigation effect in infected birds but helped the birds fed DDGS to maintain feed intake and BW gain. PMID:23687155

  10. Thumb-loops up for catalysis: a structure/function investigation of a functional loop movement in a GH11 xylanase

    Directory of Open Access Journals (Sweden)

    Thierry Siméon

    2012-07-01

    Full Text Available Dynamics is a key feature of enzyme catalysis. Unfortunately, current experimental and computational techniques do not yet provide a comprehensive understanding and description of functional macromolecular motions. In this work, we have extended a novel computational technique, which combines molecular modeling methods and robotics algorithms, to investigate functional motions of protein loops. This new approach has been applied to study the functional importance of the so-called thumb-loop in the glycoside hydrolase family 11 xylanase from Thermobacillus xylanilyticus (Tx-xyl. The results obtained provide new insight into the role of the loop in the glycosylation/deglycosylation catalytic cycle, and underline the key importance of the nature of the residue located at the tip of the thumb-loop. The effect of mutations predicted in silico has been validated by in vitro site-directed mutagenesis experiments. Overall, we propose a comprehensive model of Tx-xyl catalysis in terms of substrate and product dynamics by identifying the action of the thumb-loop motion during catalysis.

  11. A 24.7-kDa copper-containing oxidase, secreted by Thermobifida fusca, significantly increasing the xylanase/cellulase-catalyzed hydrolysis of sugarcane bagasse.

    Science.gov (United States)

    Chen, Cheng-Yu; Hsieh, Zhi-Shen; Cheepudom, Jatuporn; Yang, Chao-Hsun; Meng, Menghsiao

    2013-10-01

    Thermobifida fusca is a moderately thermophilic soil bacterium belonging to Actinobacteria. It has been known for its capability to degrade plant cell wall polymers except lignin and pectin. To know whether it can produce enzymes to facilitate lignin degradation, the extracellular proteins bound to sugarcane bagasse were harvested and identified by liquid chromatography tandem mass spectrometry. Among the identified proteins, a putative copper-containing polyphenol oxidase of 241 amino acids, encoded by the locus Tfu_1114, was thought to presumably play a role in lignin degradation. This protein (Tfu1114) was thus expressed in E. coli and characterized. Similarly to common laccases, Tfu1114 is able to catalyze the oxidation reaction of phenolic and nonphenolic lignin related compounds such as 2,6-dimethoxyphenol and veratryl alcohol. More interestingly, it can significantly enhance the enzymatic hydrolysis of bagasse by xylanase and cellulase. Tfu1114 is stable against heat, with a half-life of 4.7 h at 90 °C, and organic solvents. It is sensitive to ethylenediaminetetraacetic acid and reducing agents but resistant to sodium azide, a potent inhibitor of laccases. Atomic absorption spectroscopy indicated that the ratio of copper to the protein monomer is 1, instead of 4, a feature of classical laccases. All these data suggest that Tfu1114 is a novel oxidase with laccase-like activity, potentially useful in biotechnology application. PMID:23377789

  12. Influence of xylanase addition on the characteristics of loaf bread prepared with white flour or whole grain wheat flour Influência da adição de xilanase nas características de pão de forma preparado com farinha de trigo comum ou farinha de trigo de grão inteiro

    OpenAIRE

    Leandra Zafalon Jaekel; Camila Batista da Silva; Caroline Joy Steel; Yoon Kil Chang

    2012-01-01

    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 form...

  13. Insight into structure and assembly of the nuclear pore complex by utilizing the genome of a eukaryotic thermophile

    DEFF Research Database (Denmark)

    Amlacher, Stefan; Sarges, Phillip; Flemming, Dirk;

    2011-01-01

    Despite decades of research, the structure and assembly of the nuclear pore complex (NPC), which is composed of ~30 nucleoporins (Nups), remain elusive. Here, we report the genome of the thermophilic fungus Chaetomium thermophilum (ct) and identify the complete repertoire of Nups therein. The...... thermophilic proteins show improved properties for structural and biochemical studies compared to their mesophilic counterparts, and purified ctNups enabled the reconstitution of the inner pore ring module that spans the width of the NPC from the anchoring membrane to the central transport channel. This module...

  14. Selection of Bacillus spp. for Cellulase and Xylanase Production as Direct-Fed Microbials to Reduce Digesta Viscosity and Clostridium perfringens Proliferation Using an in vitro Digestive Model in Different Poultry Diets

    Science.gov (United States)

    Latorre, Juan D.; Hernandez-Velasco, Xochitl; Kuttappan, Vivek A.; Wolfenden, Ross E.; Vicente, Jose L.; Wolfenden, Amanda D.; Bielke, Lisa R.; Prado-Rebolledo, Omar F.; Morales, Eduardo; Hargis, Billy M.; Tellez, Guillermo

    2015-01-01

    Previously, our laboratory has screened and identified Bacillus spp. isolates as direct-fed microbials (DFM). The purpose of the present study was to evaluate the cellulase and xylanase production of these isolates and select the most appropriate Bacillus spp. candidates for DFM. Furthermore, an in vitro digestive model, simulating different compartments of the gastrointestinal tract, was used to determine the effect of these selected candidates on digesta viscosity and Clostridium perfringens proliferation in different poultry diets. Production of cellulase and xylanase were based on their relative enzyme activity. Analysis of 16S rRNA sequence classified two strains as Bacillus amyloliquefaciens and one of the strains as Bacillus subtilis. The DFM was included at a concentration of 108 spores/g of feed in five different sterile soybean-based diets containing corn, wheat, rye, barley, or oat. After digestion time, supernatants from different diets were collected to measure viscosity, and C. perfringens proliferation. Additionally, from each in vitro simulated compartment, samples were taken to enumerate viable Bacillus spores using a plate count method after heat-treatment. Significant (P < 0.05) DFM-associated reductions in supernatant viscosity and C. perfringens proliferation were observed for all non-corn diets. These results suggest that antinutritional factors, such as non-starch polysaccharides from different cereals, can enhance viscosity and C. perfringens growth. Remarkably, dietary inclusion of the DFM that produce cellulase and xylanase reduced both viscosity and C. perfringens proliferation compared with control diets. Regardless of diet composition, 90% of the DFM spores germinated during the first 30 min in the crop compartment of the digestion model, followed by a noteworthy increased in the intestine compartment by ~2log10, suggesting a full-life cycle development. Further studies to evaluate in vivo necrotic enteritis effects are in

  15. ENZYMATIC PRETREATMENT WITH XYLANASE FOR IMPROVING BLEACHABILITY AND BRIGHTNESS OF WHEAT STRAW CHEMOMECHANICAL PULP%木聚糖酶预处理对麦草化学机械浆可漂性及白度的改善

    Institute of Scientific and Technical Information of China (English)

    洪枫; 刘明山; 房桂干; 沈兆邦

    2001-01-01

    Xylanase from Trichoderma ressei Rut C-30 with corncob meal ascarbon source was prepared and improvement of bleachability and brightness of wheat straw CMP by xylanase pretreatment were investigated. The results indicated that the activity of xylanase was up to 38.34IU/mL with 18g/L(oven dry weight)corncob as the substrate. The xylanase treatment was beneficial to improving the bleachability of wheat straw CMP, enhancing hydrogen peroxide bleaching, increasing the brightness effectively and decreasing the consumption of bleaching agent. The research showed that 50% hydrogen peroxide was saved after enzymic pretreatment when wheat straw CMP was bleached to the same brightness with one-stage H2O2. Furthermore if wheat straw CMP was bleached with two-stage high consistence hydrogen peroxide after enzymatic pretreatment, namely XP3P3(total H2O2 6%)bleaching sequence, the brightness could be over 60%(ISO).%探讨了以玉米芯为碳源制备木聚糖酶及麦草化学机械浆经该木聚糖酶预处理后可漂性和白度的改善效果。结果表明,直接以玉米芯为底物、里氏木霉为菌种产酶效果较好,当底物浓度为18g/L时,木聚糖酶活力可达38.34IU/mL。木聚糖酶预处理有利于改善麦草化机浆的可漂性,促进其过氧化氢漂白,有效提高漂白浆白度,降低漂剂消耗。研究表明,当经单段H2O2漂至相同白度时,木聚糖酶预处理后可节约50%的H2O2用量。若麦草CMP酶处理后采用高浓两段过氧化氢漂白,即XP3P3漂序(H2O2总量为6%)时,白度可达60%(ISO)以上。

  16. Enzymatic Hydrolysis of Wheat Arabinoxylan by a Recombinant "Minimal" Enzyme Cocktail Containing beta-Xylosidase and Novel endo-1,4-beta-Xylanase and alpha-L-Arabinofuranosidase Activities

    DEFF Research Database (Denmark)

    Sørensen, Hanne R.; Pedersen, Sven; Jørgensen, Christel T.; Meyer, Anne Boye Strunge

    2007-01-01

    -xylanase from H. insolens (Xyl III), and a GH3 beta-xylosidase from Trichoderma reesei (beta-xyl) released 322 mg of arabinose and 512 mg of xylose per gram of water-soluble wheat arabinoxylan dry matter and 150 mg of arabinose and 266 mg of xylose per gram of water-insoluble wheat arabinoxylan dry matter after...... wheat arabinoxylan substrates were hydrolyzed with a combination of Ultraflo L and Celluclast 1.5 L, two commercially available enzyme preparations produced by H. insolens and T. reesei....

  17. Energetic values and performace of broilers feeding sorghum and soybean meal based diets supplemented with B-glucanase and B-xylanase

    Directory of Open Access Journals (Sweden)

    Evandro de Abreu Fernandes

    2016-04-01

    Full Text Available Grains, brans, and vegetable meals may contain non-starch polysaccharides (NSP, which increases viscosity in the gastrointestinal tract (GIT and interfere with the digestion and absorption of nutrients. This study aimed to evaluate the performance and determine the metabolizable energy of a sorghum-based broiler diet with and without the supplementation of an enzymatic complex. The experiments were conducted in a completely randomized design with 1200 chickens, using sorghum-based feed with and without the addition of 50 g of enzyme-CCE complex (?-glucanase and ?-xylanase, and with two levels of metabolizable energy (ME kg-1: ME; ME + CCE; reduced ME (-50 kcal kg-1; and reduced ME + CCE. The data were subjected to an analysis of variance and the means were compared using a Tukey’s test at the 5% significance level. At 42 and 47 days of age, the living weight of the birds fed with the reduced ME was low, while birds fed with reduced ME + CCE had the same weight as those fed with other energy diets (ME and ME + CCE. Feed conversion was poorest at 47 days of age for the birds on reduced ME diet. In the metabolic test (with fattening diets to determine AME and AMEn, the reduced ME diet had the lowest result, confirming the effect of the addition of enzymes. The addition of CCE to sorghum-based diets provides enough enzymatic activity to increase the metabolizable energy of the diet (50 kcal of AME and influence the growth performance of broilers at the slaughtering age.

  18. Scientific Opinion on the safety and efficacy of Feedlyve AXC (endo-1,4-beta-xylanase as a feed additive for turkeys

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed

    2012-07-01

    Full Text Available

    Feedlyve AXC is an enzyme preparation containing an endo-1,4-beta-xylanase produced by a non-genetically modified strain of Trichoderma koningii. The additive is intended for use in turkeys and is presented in several forms. A tolerance study performed with a 100-fold of the maximum recommended dose (100 AXC/kg feed showed no adverse effects in turkeys for fattening. Therefore, it is concluded that Feedlyve AXC is safe for turkeys for fattening at the maximum recommended dose. This conclusion is extended to turkeys reared for breeding. The results of two genotoxicity in vitro studies and a sub-chronic oral toxicity study did not indicate any concern for consumer safety arising from the use of the additive in feed for turkeys. Feedlyve AXC 6000P was negative in dermal and eye irritation tests in the rabbit. No data on the skin/eye irritation potential of the BRUTE formulation is available. Therefore, the FEEDAP Panel considers it prudent to treat this formulation as a potential skin/eye irritant. No skin sensitization and no acute respiratory toxicity were observed in appropriate tests using the BRUTE formulation. Due to the proteinaceous nature of the compound the additive is to be considered as a potential respiratory sensitiser. The active substance of Feedlyve AXC is a protein and as such will be degraded/inactivated during the passage through the digestive tract of animals. Therefore, no risks for the environment are expected and no further environmental risk assessment is required. A total of three efficacy studies in turkeys for fattening were provided. Results showed that the additive has the potential to be efficacious when added to diets for turkeys for fattening at a minimum dose of 75 AXC/kg complete feed. This conclusion can be extended to turkeys reared for breeding.

  19. Scientific Opinion on the safety and efficacy of Hostazym X (endo-1,4-beta-xylanase as a feed additive for poultry, piglets and pigs for fattening

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2013-02-01

    Full Text Available Hostazym X is an enzyme preparation of xylanase produced by a non-genetically modified strain of Trichoderma citrinoviride. The fermentation product showed negative results in a bacterial reverse mutation assay. The results of an in vitro chromosomal aberration test and of an in vivo comet assay indicate the presence of genotoxic activity in the product. Although the tolerance studies provided in target species did not indicate any adverse effect of the additive, the FEEDAP Panel, taking into account the genotoxic hazard from the product, cannot conclude on the safety of the additive for the target species. The results obtained in a subchronic rat study did not indicate any concerns for consumer safety. However, owing to the lack of information on the nature and fate of potentially genotoxic material in food derived from animals receiving the additive, the FEEDAP Panel cannot conclude on the safety of the additive for the consumer. The product should be considered a potential skin and eye irritant, and a potential skin and respiratory sensitiser. Owing to the presence of genotoxic activity in the product, any level of exposure to the additive is considered hazardous. No risks to the environment are expected to result from the use of the additive in feed, and, therefore, no further environmental risk assessment is required. Based on the results obtained in the efficacy studies, the FEEDAP Panel concludes that the additive has the potential to be efficacious in turkeys for fattening at the dose of 1 050 EPU/kg feed and in chickens for fattening, laying hens, piglets (weaned and pigs for fattening at the dose of 1 500 EPU/kg feed. Conclusions on the efficacy can be extrapolated to minor poultry species.

  20. 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.

  1. The critical role of partially exposed N-terminal valine residue in stabilizing GH10 xylanase from Bacillus sp.NG-27 under poly-extreme conditions.

    Directory of Open Access Journals (Sweden)

    Amit Bhardwaj

    Full Text Available BACKGROUND: Understanding the mechanisms that govern protein stability under poly-extreme conditions continues to be a major challenge. Xylanase (BSX from Bacillus sp. NG-27, which has a TIM-barrel structure, shows optimum activity at high temperature and alkaline pH, and is resistant to denaturation by SDS and degradation by proteinase K. A comparative circular dichroism analysis was performed on native BSX and a recombinant BSX (R-BSX with just one additional methionine resulting from the start codon. The results of this analysis revealed the role of the partially exposed N-terminus in the unfolding of BSX in response to an increase in temperature. METHODOLOGY: We investigated the poly-extremophilicity of BSX to deduce the structural features responsible for its stability under one set of conditions, in order to gain information about its stability in other extreme conditions. To systematically address the role of the partially exposed N-terminus in BSX stability, a series of mutants was generated in which the first hydrophobic residue, valine (Val1, was either deleted or substituted with various amino acids. Each mutant was subsequently analyzed for its thermal, SDS and proteinase K stability in comparison to native BSX. CONCLUSIONS: A single conversion of Val1 to glycine (Gly changed R-BSX from being thermo- and alkali- stable and proteinase K and SDS resistant, to being thermolabile and proteinase K-, alkali- and SDS- sensitive. This result provided insight into the structure-function relationships of BSX under poly-extreme conditions. Molecular, biochemical and structural data revealed that the poly-extremophilicity of BSX is governed by a partially exposed N-terminus through hydrophobic interactions. Such hitherto unidentified N-terminal hydrophobic interactions may play a similar role in other proteins, especially those with TIM-barrel structures. The results of the present study are therefore of major significance for protein folding

  2. 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)

    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

  3. Scientific Opinion on the efficacy of Natugrain® TS/TS L (endo-1,4-beta-xylanase and endo-1,4-beta-glucanase as a feed additive for laying hens

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2014-06-01

    Full Text Available Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP was asked to deliver a scientific opinion on the efficacy of Natugrain® TS/TS L in laying hens. This feed additive contains endo-1,4-beta-xylanase and endo-1,4-beta-glucanase, produced by two genetically modified strains of Aspergillus niger. The product is available in solid (Natugrain® TS and liquid (Natugrain® TS L forms. This additive is authorised for use in weaned piglets and pigs for fattening, poultry species and ornamental birds. The applicant is now seeking a modification of the terms of the authorisation to reduce the recommended dose in laying hens from 560 thermostable xylanase units (TXU and 250 thermostable glucanase units (TGU per kg feed to 280 TXU and 125 TGU per kg feed. The FEEDAP Panel assumes that the two formulations of the additive are equivalent in terms of efficacy. A total of three trials were provided. Based on the results obtained, the Panel concludes that Natugrain® TS has the potential to be efficacious in laying hens at the dose of 280 TXU and 125 TGU per kg feed.

  4. Cloning and Sequencing of Xylanase cDNA from Volvariella volvacea Using Conserved Sequences in Cellulose-Binding Domain%利用真菌纤维素结合域(CBD)保守性序列进行草菇木聚糖酶cDNA的克隆

    Institute of Scientific and Technical Information of China (English)

    丁少军; J.A.BUSWELL

    2004-01-01

    Cellulose-binding domains (CBDs) are present in the majority of fungal cellulases and hemicellulases. Based on the conserved region of CBDs, degenerate primers were designed and used to amplify the 5′-end cDNA fragment of xylanase from Volvariella volvacea by 5′-RACE. Gene specific primer was then designed based on extreme region of 5′-end cDNA fragment and used to amplify the full length cDNA of xylanase. The cDNA of xynl was 1287 bp in length, including 3′and 5′-non-coding region. The xynl cDNA contained an ORF of 1101 bp encoding 367 amino acids, in which there was a putative signal peptide with 19 amino acids. Alignment of the deduced amino acid sequence of xynl with other xylanases showed that the homology with family-10 xylanases from Agaricus bisporus xyll, AspergiUus sojae xynl, Aspergillus kawachii xynA, Fusarium oxysporumf, sp xyl3 was 64% ,55% ,52% ,55% , respectively.

  5. A computational method for the systematic screening of reaction barriers in enzymes: searching for Bacillus circulans xylanase mutants with greater activity towards a synthetic substrate

    Directory of Open Access Journals (Sweden)

    Martin R. Hediger

    2013-07-01

    Full Text Available We present a semi-empirical (PM6-based computational method for systematically estimating the effect of all possible single mutants, within a certain radius of the active site, on the barrier height of an enzymatic reaction. The intent of this method is not a quantitative prediction of the barrier heights, but rather to identify promising mutants for further computational or experimental study. The method is applied to identify promising single and double mutants of Bacillus circulans xylanase (BCX with increased hydrolytic activity for the artificial substrate ortho-nitrophenyl β-xylobioside (ONPX2. The estimated reaction barrier for wild-type (WT BCX is 18.5 kcal/mol, which is in good agreement with the experimental activation free energy value of 17.0 kcal/mol extracted from the observed kcat using transition state theory (Joshi et al., 2001. The PM6 reaction profiles for eight single point mutations are recomputed using FMO-MP2/PCM/6-31G(d single points. PM6 predicts an increase in barrier height for all eight mutants while FMO predicts an increase for six of the eight mutants. Both methods predict that the largest change in barrier occurs for N35F, where PM6 and FMO predict a 9.0 and 15.8 kcal/mol increase, respectively. We thus conclude that PM6 is sufficiently accurate to identify promising mutants for further study. We prepared a set of all theoretically possible (342 single mutants in which every amino acid of the active site (except for the catalytically active residues E78 and E172 was mutated to every other amino acid. Based on results from the single mutants we construct a set of 111 double mutants consisting of all possible pairs of single mutants with the lowest barrier for a particular position and compute their reaction profile. None of the mutants have, to our knowledge, been prepared experimentally and therefore present experimentally testable predictions.

  6. FERROFLUIDS INFLUENCE ON DEHYDROGENASES ACTIVITY IN CELLULOLYTIC FUNGUS CHAETOMIUM GLOBOSUM

    Directory of Open Access Journals (Sweden)

    Alexandru Manoliu

    2003-08-01

    Different results were noticed for different ferrofluids concentrations: 20, 40, 60, 80 and 100 μl/L. Inhibitory or stimulatory ferrofluids effect was obtained depending on the nature of the investigated enzyme.

  7. Purification and characterization of a new Xylanase from Humicola grisea var. Thermoidea Produção, purificação e caracterização de uma nova Xilanase de Humicola grisea var. Thermoidea

    Directory of Open Access Journals (Sweden)

    Severino de Albuquerque Lucena-Neto

    2004-06-01

    Full Text Available The thermophilic fungus Humicola grisea var. thermoidea secretes extracellular xylanase when grown on solid and in liquid media containing wheat bran and banana plant residue as substrates, respectively. At 55ºC, xylanase from the culture filtrate of H. grisea var. thermoidea grown on banana stalk retained 50% of its activity after 28 h of incubation. A xylanase (X2 was isolated from solid state cultures with wheat bran as the carbon source. It was purified to apparent homogeneity by ultrafiltration followed by ion-exchange and hydrophobic interaction chromatography on DEAE-Sepharose and Phenyl-Sepharose resins, respectively. The enzyme had an apparent molecular weight of 29 kDa, as determined by SDS-PAGE. The purified enzyme was most active at pH and temperature ranges of 4.5-6.5 and 55-60ºC, respectively. In addition, X2 showed thermostability at 60ºC with a half-life of approx. 5.5 h. The apparent Km values, using soluble and insoluble arabinoxylans as substrates, were 10.87 and 11.20 mg/ml, respectively.O fungo termofílico Humicola grisea var. secreta xilanase extracelular quando cultivado em meios sólidos e líquidos contendo farelo de trigo e engaço de bananeira como substratos, respectivamente. À temperatura de 55ºC, xilanase do filtrado de meio de cultura de H. grisea var. thermoidea cultivado em engaço de bananeira reteve 50% de sua atividade após 28 de incubação. Uma xilanase (X2 foi isolada de culturas de estado sólido contendo farelo de trigo como fonte de carbono. X2 foi purificada por ultrafiltração, seguido por cromatografias de interação hidrofóbica e troca iônica em resinas de Phenyl-Sepharose e DEAE-Sepharose, respectivamente. A enzima apresentou peso molecular de 29 kDa, como determinado por SDS-PAGE. A enzima purificada foi mais ativa em intervalos de pH e temperatura de 4,5-6,5 and 55-60ºC, respectivamente. Além disso, X2 mostrou termoestabilidade a 60ºC com meia vida de aproximadamente 5,5 h. Os

  8. Production and properties of the cellulase-free xylanase from Thermomyces lanuginosus IOC-4145 Produção e propriedades de xilanase livre de celulase de Thermomyces lanuginosus IOC-4145

    Directory of Open Access Journals (Sweden)

    Mônica Caramez Triches Damaso

    2002-12-01

    Full Text Available In recent years, xylanases have expanded their use in many processing industries, such as pulp and paper, food and textile. Thermomyces lanuginosus IOC-4145 was able to produce a very high level of cellulase-free xylanase in shaken cultures using corncob as substrate (500 U/mL. An optimization of the medium composition in submerged fermentation was carried out aiming at a low cost medium composition for enzyme production. Statistical experiment design was employed for this purpose, pointing out corncob as the most important parameter, which affects enzyme production. Additionally, the influence of several chemicals on xylanase activity was investigated in the crude extract. A slight stimulation of the enzyme (5-15% was achieved with NaCl and urea, both at 3 and 5 mM of concentration. On the other hand, dithiothreitol and beta-mercaptoethanol at a molarity of 5mM have caused a strong stimulation of the enzyme (40-53%. The crude xylanase displayed appreciable thermostability, retaining almost 50% of activity during 24 hours of incubation at 50ºC; about 50% of activity was present at 60ºC even after 4 hours of incubation. The enzyme also exhibited good storage stability at -20ºC without any stabilizing agent.Nos últimos anos tem crescido o uso de xilanases em muitas indústrias, tais como polpa e papel, alimentos e têxtil. Thermomyces lanuginosus IOC-4145 foi capaz de produzir um alto nível de xilanase livre de celulase em culturas agitadas usando sabugo de milho como substrato (500 U/mL. Procedeu-se, inicialmente, à otimização da composição do meio de produção em fermentação submersa, com o intuito de alcançar uma composição de meio de produção de baixo custo para produção da enzima. Para este propósito, utilizou-se planejamento estatístico de experimentos. O sabugo de milho revelou-se como a mais importante variável que afeta a produção enzimática. Adicionalmente, a influência de vários reagentes na atividade xilan

  9. Coexpression of β-mannanase Gene and Xylanase Gene in Pichia pastoris%β-甘露聚糖酶基因和木聚糖酶基因在毕赤母中的共表达

    Institute of Scientific and Technical Information of China (English)

    李剑芳; 赵顺阁; 邬敏辰; 张慧敏; 魏喜换

    2012-01-01

    β -mannanase and xylanase are extensively used in food industry. Synergistic action between the two enzymes is obersved to effectively improve digestibility of the hemicelluloses of food material. In order to coexpress β -mannanase gene and xylanase gene in Pichia pastoris, pPIC9K-xyn Ⅱ was linearized with Sal /and transformed into GSll5/Anman5A by electroporation. GS115/A nman5A-xyn Ⅱ with high production of β-mannanase and xylanase was screened by using G418. SDS-PAGE demonstrated two protein bands with apparent molecular weight of about 52 000 and 24 100. One GS115/Anman5A-xyn Ⅱ transformant producing the highest activities of two enzymes in shake flasks was selected and numbered as GSMX2. Subsequently the expression conditions for enzymes production by using GSMX2 were optimized, the optimum expression conditions listede as follows:initial pH 7.0,glycerol concentration 1.5% ,methanol concentration 1.5%, induction for 120 h. Under these conditions, the activities of β-mannanase and xylanase were achieved at 37.1 U/mL and 193.6 U/mL, respectively.%为实现β-甘露聚糖酶基因和木聚糖酶基因在毕赤酵母中的共表达,作者将经SalⅠ线性化的pPIC9K-xynⅡ电转化至工程菌GS115/Anman5A中,经G418浓度梯度筛选后获得能同时高产β-甘露聚糖酶和木聚糖酶的双重重组子GS115/Anman5A-xynⅡ.SDS-PAGE显示目的蛋白的相对分子质量分别约为52 000,24 100.摇瓶发酵筛选出产酶活性最强的转化株,命名为GSM X2,随后对该菌株的表达条件进行初步优化,优化后的表达条件为:诱导时间120 h,培养基起始pH值为7.0,甘油添加量为1.5%,甲醇添加量为1.5%.在此培养条件下,产β-甘露聚糖酶和木聚糖酶的活性分别达到37.1 U/mL和193.6 U/mL.

  10. Scientific opinion on the safety and efficacy of Natugrain® TS (endo-1,4-beta-xylanase and endo-1,4-beta-glucanase as a feed additive for pigs for fattening

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2013-07-01

    Full Text Available Natugrain® TS is an enzyme preparation that contains endo-1,4-beta-xylanase and endo-1,4-beta-glucanase, produced by two genetically modified strains of Aspergillus niger. This product is currently authorised for use in piglets (weaned, poultry species and ornamental birds as a zootechnical additive under the functional group of digestibility enhancers. The applicant is now seeking for the extension of its use as a zootechnical additive for pigs for fattening at a dose range of 560 to 840 TXU (thermostable xylanase units and 250 to 375 TGU (thermostable glucanase units per kg feed. Since the product has been demonstrated to be  safe in piglets at the maximum recommended dose, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP considers that this conclusion can be extended to pigs for fattening, provided that the same maximum dose applies. In two long-term trials, Natugrain® TS significantly improved the performance of the pigs for fattening at the minimum recommended dose (560 TXU and 250 TGU/kg feed. In a balance trial, the metabolisable energy content of the diet was significantly improved by the addition of Natugrain® TS in feed at the minimum recommended dose. Therefore, the FEEDAP Panel concludes that Natugrain® TS at the minimum recommended dose (560 TXU and 250 TGU/kg feed has the potential to be efficacious in pigs for fattening.

  11. Scientific Opinion on the safety and efficacy of AveMix® XG 10 (endo-1,4-beta-xylanase and endo-1,3(4-beta-glucanase as a feed additive for pigs for fattening and minor porcine species

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    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2013-04-01

    Full Text Available The additive AveMix® XG 10 is an enzyme preparation of endo-1,4-beta-xylanase (xylanase and endo-1,3(4-beta-glucanase (glucanase, produced by two strains of Trichoderma reesei. This product is currently authorised for use in chickens for fattening, laying hens, minor poultry species and weaned piglets as a zootechnical additive, under the functional group of digestibility enhancers. The applicant is now seeking an extension of the authorisation to pigs for fattening and minor porcine species at a recommended dose of 4 000 XU (xylanase units and 900 BGU (beta-glucanase units per kg complete feed. The FEEDAP Panel considers that, since the additive has been demonstrated to be safe for piglets at the recommended dose, this conclusion can be extended to pigs for fattening and extrapolated to minor growing porcine species. Supplementation of diets for pigs for fattening with Avemix® XG 10 at the recommended dose (4 000 XU and 900 BGU/kg resulted in a significantly higher body weight and daily weight gain in one trial and significantly improved feed to gain ratio in two trials. Therefore, the FEEDAP Panel concludes that AveMix® XG 10 has the potential to be efficacious in pigs for fattening at this dose. Since the mode of action of xylanases and glucanases can reasonably be assumed to be the same in all porcine species, the conclusions on efficacy for weaned piglets and pigs for fattening can be extrapolated to include all minor porcine species for growing at 4 000 XU and 900 BGU/kg complete feed.

  12. Expression of cold-adapted β-1,3-xylanase as a fusion protein with a ProS2 tag and purification using immobilized metal affinity chromatography with a high concentration of ArgHCl.

    Science.gov (United States)

    Kudou, Motonori; Okazaki, Fumiyoshi; Asai-Nakashima, Nanami; Ogino, Chiaki; Kondo, Akihiko

    2015-01-01

    Cold-adapted β-1,3-xylanase (P.t.Xyn26A) from the psychrotrophic bacterium, Psychroflexus torquis, was expressed as a fusion protein with tandem repeats of the N-terminal domain of Protein S from Myxocuccus xanthus (ProS2) in Escherichia coli. After cell lysis in phosphate buffer, most of the ProS2-P.t.Xyn26A was located in the insoluble fraction and aggregated during purification. Arginine hydrochloride (ArgHCl) efficiently solubilized the ProS2-P.t.Xyn26A. The solubilized ProS2-P.t.Xyn26A was purified using immobilized metal affinity chromatography (IMAC) with 500 mM ArgHCl. After cleavage of ProS2-P.t.Xyn26A by human rhinovirus 3C protease, we confirmed that recombinant P.t.Xyn26A maintained its native fold. This is the first report of the expression of a cold-adapted enzyme fused with a ProS2 tag under IMAC purification using a high concentration of ArgHCl. These insights into the expression and purification should be useful during the handling of cold-adapted enzymes. PMID:25214227

  13. Xylanase production with xylan rich lignocellulosic wastes by a local soil isolate of Trichoderma viride Produção de xilanase com resíduos lignocelulósicos ricos em xilana por uma cepa local de Trichoderma viride isolada de solo

    Directory of Open Access Journals (Sweden)

    Meenakshi Goyal

    2008-09-01

    Full Text Available In the present study, cultural and nutritional conditions for enhanced production of xylanase by a local soil isolate of Trichoderma viride, using various lignocellulosic substrates in submerged culture fermentation have been optimized. Of the lignocellulosics used, maize straw was the best inducer followed by jowar straw for xylanase production. The highest activity achieved was between 14 to 17 days of fermentation. A continuous increase in xylanase production was observed with increasing level of lignocellulosics in the medium and highest activity was observed with maize straw at 5% level. Xylanase production with higher levels of lignocellulosics (3 to 5% of maize, jowar and barseem was found to be higher as compared to that with commercial xylan as carbon source. Sodium nitrate was the best nitrogen source among the six sources used. Maximum xylanase production was achieved with initial medium pH of 3.5-4.0 and incubation temperature of 25ºC.The enzyme preparation was effective in bringing about saccharification of different lignocellulosics. The xylanase production could be further improved by using alkali treated straw as carbon source.Neste estudo, otimizou-se as condições culturais e nutricionais para produção aumentada de xilanase por uma cepa local de Trichoderma viride isolada de solo, empregando-se vários substratos lignocelulósicos, em fermentação submersa. Entre os substratos utilizados, o melhor indutor de produção de xilanase foi palha de milho, seguido de palha de sorgo. A atividade mais alta foi obtida entre 14 e 17 dias de fermentação. Com palha de milho observou-se um aumento contínuo na produção de xilanase com o aumento da concentração dos substratos lignocelulósicos no meio, sendo que a melhor atividade foi obtida com 5% de palha de milho. A produção de xilanase com níveis mais altos de (3 a 5% de milho, sorgo e forragem verde (barseem foi mais levada do que com xilana comercial como fonte de carbono

  14. 产木聚糖酶的沿海红树林真菌筛选及其培养与酶活测定条件优化%Survey of coastal mangrove fungi for xylanase production and optimized culture and assay conditions

    Institute of Scientific and Technical Information of China (English)

    袁康培; 关利平; 冯明光

    2005-01-01

    从香港海岸红树林分离到的77株真菌中有34株可产生木聚糖酶,从中选出CY2809(Staganospora sp.)、CY4786和CY5040等3菌株与已知陆生产酶菌株HU5048(Aspergillus awamori)进行产木聚糖酶的比较研究.根据培养液中菌丝生物量、木聚糖酶活力和木糖等价还原糖含量等指标的测定,菌株CY4786在起始pH 7.8的木聚糖-酵母膏-海盐液体培养基中25℃下震荡(100r/min)培养7d产酶最佳;粗酶液在50q℃和pH 4.6的优化条件下进行测定,木聚糖酶活力达到1.07×104U/mL.结果表明,红树林真菌起着半纤维素降解者的作用,沿海红树林环境中存在着可资利用的木聚糖酶产生菌.作者讨论了利用发酵液中木糖等价还原糖含量的动态变化作为快速筛选产木聚糖酶菌株的指标的可能性.%Xylanase activity was detected among 34 of 77 fungal isolates derived from decaying wood, debris and soil samples collected in coastal mangrove environment of Hong Kong. Of those, three isolates CY2809 (Staganospora sp. ), CY4786 and CY5040 were chosen for comparison of xylanase production in parallel to HU5048 ( Aspergillus awamori ), a terrestrial, highly productive isolate. Based on the assessment of mycelial biomass, xylanase activity and content of xylose-equivalent reducing sugars in their liquid cultures, the isolate CY4786 was best for xylanase production in a basal medium containing birchwood xylan (10.0 g/L)as a sole carbon source, yeast extract (2.5 g/L) and sea salts (15.0 g/L) with initial pH 7.8. When assayed at the optimized regime of 50℃ and pH 4.6, the activity of xylanase produced by CY4786 in 7d liquid culture at 25℃ reached 1.07 × 104 unit/mL. The results indicate that the mangrove fungi act as hemicellulose decomposers in the mangrove environment where highly xylanaseproductive isolates can be searched for exploitation. A discussion is given on the possible use of the content of xylose-equivalent reducing sugars as an index to

  15. Tropical Soil Fungi Producing Cellulase and Related Enzymes in Biodegradation

    Directory of Open Access Journals (Sweden)

    Wattanachai Pathomsiriwong

    2012-01-01

    Full Text Available The objective of this study was to screen, identify and characterize of cellulolytic fungi from various soil management fields; organic, young organic, semi-chemical and chemical soil from Surin rice fields, Thailand. Fungi from various type of soils were isolated by dilution plating technique on Reese minimal medium supplemented with cellulose powder and rice straw then incubated at 25°C for 3-7 days. The isolated fungi were screened and identified using slide culture technique. The enzymatic activities were assessed by qualitative method for cellulase, xylanase, peroxidase and laccase activities. Two-hundred and fifty-eight fungi isolates found in Surin rice fields belonging to the genus Penicillium (5 species, Paecilomyces (4 species, Aspergillus (3 species, Acremonium (2 species, Chaetomium (2 species, Alternaria (1 species, Bipolaris (1 species, Curvularia (1 species, Fusarium (1 species, Humicola (1 species, Mucor (1 species, Nigrospora (1 species, Phoma (1 species, Pyrenochaeta (1 species, Pythium (1 species, Rhizopus (1 species, Sporotrichum (1 species and Trichoderma (1 species. Out of 29 fungal species clearly showed different in enzymatic activities. Most tropical soil fungi had ability to produce cellulase, xylanase, laccase and peroxidase, respectively. The highest capacity was found only in cellulase. Aspergillus niger, Aspergillus sp., Chaetomium murorum and Trichoderma sp. showed the highest potential to produce cellulase. Eleven species of soil fungi showed high capacity in xylanase activity. For laccase and peroxidase activity, there were found in 2 species. The results also revealed that only ten showed highest carboxymethyl cellulase, xylanase, peroxidase and laccase activities by qualitative screening method for enzymatic assay. They were identified as Aspergillus niger, Acremonium sp., Aspergillus sp., Chaetomium murorum, Humicola grisea, Mucor sp., Paecilomyces victoriae, Penicillium janthinellum, Penicillium lanosum and

  16. Scientific opinion on the safety and efficacy of Ronozyme WX (endo-1,4-beta-xylanase as a feed additive for poultry, piglets (weaned and pigs for fattening

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed

    2012-07-01

    Full Text Available

    Ronozyme WX is a preparation of endo-1,4-beta-xylanase produced by a genetically modified strain of Aspergillus oryzae and is presented in two forms, coated (CT and liquid (L. The solid and liquid forms of the additive are considered equivalent in terms of safety and efficacy at the same dose. The final enzyme preparations contain no cultivable production organisms or recombinant DNA, given the limits of detection and evidence for the presence of deoxyribonuclease activity. The use of Ronozyme WX is safe at the maximum proposed dose for chickens and turkeys for fattening and piglets. This conclusion allows extrapolation to all minor poultry species for fattening and can be extended to pigs for fattening. Ronozyme WX is not genotoxic and no adverse effects were reported in a 90-day oral toxicity study. Therefore, the use of Ronozyme WX as a feed additive is not considered to pose a risk to consumers of animal products derived from animals treated with the additive. No studies of the additive that are relevant to user safety were carried out. However, the enzyme concentrate in Ronozyme WX was tested and was found not to be irritant to skin or eyes. In the absence of data, and given the nature of the product, it should be treated as a potential skin/respiratory sensitiser. The active ingredient of Ronozyme WX is a protein and as such will be degraded/inactivated during passage through the digestive tract of animals. Therefore, no risks to the environment are expected. The efficacy of Ronozyme WX has been demonstrated in chickens, turkeys and ducks for fattening at a minimum proposed dose of 100 FXU/kg complete feed. This conclusion can be extended to all minor poultry species for fattening. Efficacy has also been demonstrated in piglets and in pigs for fattening at a minimum dose of 200 FXU/kg complete feed.

  17. Scientific Opinion on the safety and efficacy of Safizym® X (endo-1,4-beta-xylanase as a feed additive for chickens and turkeys for fattening and laying hens

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2014-01-01

    Full Text Available Safizym® X is an enzyme preparation of endo-1,4-beta-xylanase produced by a non-genetically modified strain of Trichoderma reesei which is presented in solid and liquid forms. The solid and liquid formulations are considered to be equivalent in terms of safety and efficacy for the target species. In the tolerance studies provided, the animals tolerated well 100-fold (chickens and laying hens or 10-fold (turkeys for fattening the minimum recommended dosage. Therefore, the FEEDAP Panel concludes that the additive is safe for chickens and turkeys for fattening at 1 400 IFP/kg feed and in laying hens at 840 IFP/kg feed. The results obtained in the toxicological studies are insufficient to reach conclusions on the safety for the consumers. The identity of the test item used and its relationship with the fermentation product that is used to formulate the additive was not provided. Therefore, the FEEDAP Panel cannot conclude on the safety of the additive for the consumer. The solid final formulation is not considered a skin or eye irritant; however, the liquid formulation should be considered as a skin and eye irritant. In the absence of data on skin sensitisation, and taking account of its proteinaceous nature, the additive is to be considered a potential skin and respiratory sensitiser. The active substance of Safizym® X is a protein, and as such will be degraded/inactivated during passage through the digestive tract of animals. Therefore, no risks to the environment are expected and no further environmental risk assessment is required. The FEEDAP Panel concludes that the additive has the potential to be efficacious at a dose of 1 400 IFP/kg feed in chickens and turkeys for fattening and at 840 IFP/kg feed in laying hens.

  18. Production and characterization of an enzyme complex from a new strain of Clostridium thermocellum with emphasis on its xylanase activity Produção e caracterização de um complexo enzimático de uma nova linhagem de Clostridium thermocellum com enfase em sua atividade de xilanase

    Directory of Open Access Journals (Sweden)

    Werner Bessa Vieira

    2007-06-01

    Full Text Available A new bacterial strain (ISO II was isolated from manure cow and identified as phylogenetically close to the thermophilic cellulolytic bacterium Clostridium thermocellum. The new strain produced extracellular xylanase, pectinase, mannanase and cellulase activities when grown in liquid culture medium containing banana stem as carbon source. The enzyme production profile after growth on banana stem showed that xylanase and cellulase activities were detected in different incubation periods. An enzyme complex containing xylanase, cellulase and mannanase activities was isolated from culture supernatant samples of strainISO II. The complex was partially purified by ultrafiltration and gel filtration chromatography on Sephacryl S-300. Zymogram analysis after SDS-PAGE presented at least 05 subunits with xylanase activity. The enzyme showed single protein and xylanase activity bands after electrophoresis under non-denaturing conditions. The hydrolysis of xylan was optimal at temperature range of 55-75ºC and pH 6.0. Xylanase activity was quite stable at 65ºC, retaining 80% of its original activity after 12 h incubation. The apparent Km values, using insoluble and soluble arabinoxylans as substrates, were 1.54 and 11.53 mg/mL, respectively. Xylanase was activated by dithiothreitol, L-tryptophan and L-cysteine and strongly inhibited by N-bromosuccinimide and CoCl2. The characterization of mannanase showed Km and temperature optimum of 0.846 mg/mL and 65ºC, respectively and pH 8.0. By contrast to xylanase, it was less stable at 65ºC with half-life of 2.5 h and inhibited by dithiothreitol and Ca2+.Uma nova linhagem de bactéria (ISO II foi isolada de esterco bovino e identificada como filogeneticamente próxima à bactéria termofílica Clostridium thermocellum. A nova linhagem produziu atividades de xilanase, mananase, pectinase e celulase quando cultivada em meio de cultura líquido contendo engaço de bananeira como fonte de carbono. O perfil de produ

  19. 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.

  20. Influence of xylanase addition on the characteristics of loaf bread prepared with white flour or whole grain wheat flour Influência da adição de xilanase nas características de pão de forma preparado com farinha de trigo comum ou farinha de trigo de grão inteiro

    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 O objetivo deste estudo foi verificar a influência da adição da enzima xilanase (quatro concentrações: 0, 4, 8 e 12 g.100 kg-1 de farinha nas características de pães de forma obtidos com farinha de trigo comum ou farinha de trigo de grão inteiro. Os pães produzidos com farinha comum com adição de xilanase apresentaram maior volume específico que o controle (sem enzima, porém não diferiram significativamente (p < 0,05 dos pães com as diferentes concentrações da enzima. Todas as formulações com farinha de grão inteiro adicionadas de xilanase também apresentaram volume específico significativamente maior que o controle, sendo o maior valor encontrado na formulação com 8 g xilanase.100 kg-1 de farinha. Quanto à umidade, as diferentes concentrações da enzima apresentaram pequena diferença significativa em relação ao controle. Em geral, os pães obtidos com adição de 8 g enzima.100 kg-1 de farinha apresentaram os menores valores de firmeza, apresentando as melhores características tecnológicas.

  1. Xylanases of Anaerobic Fungus Anaeromyces mucronatus

    Czech Academy of Sciences Publication Activity Database

    Novotná, Zuzana; Procházka, J.; Šimůnek, Jiří; Fliegerová, Kateřina

    2010-01-01

    Roč. 55, č. 4 (2010), s. 363-367. ISSN 0015-5632 R&D Projects: GA ČR GD525/08/H060 Institutional research plan: CEZ:AV0Z50450515 Keywords : NEOCALLIMASTIX-FRONTALIS * XYLANOLYTIC ENZYMES * POLYCENTRIC FUNGI Subject RIV: GM - Food Processing Impact factor: 0.977, year: 2010

  2. 甲酸与纤维素酶和木聚糖酶对多花黑麦草与白三叶混合青贮料发酵品质的影响%Effects of formic acid, cellulase and xylanase on fermentation quality of Lolium multiflorum and Trifolium repens mixture silage during ensiling

    Institute of Scientific and Technical Information of China (English)

    庄苏; 丁立人; 周建国; 王恬

    2013-01-01

    To evaluate the effects of formic acid, cellulase and xylanase on fermentation quality of mixture silage during ensiling, 2 000 g chopped Lolium multijlorum(80% ) and Trifolium repens(20% ) mixtures were ensiled in laboratory plastic bag either untreated (control) or treated with formic acid, cellulase, formic acid plus cellulase, xylanase, formic acid plus xylanase, cellulase plus xylanase and formic acid plus cellulase plus xylanase, respectively. Triplicate bags were opened at 0 d,2d,4d,6d,8d and 30 d of ensiling for chemical analyses. The pH value in all treated silages was lower (P<0. 05) than control at the end of ensiling. The formic acid, enzyme or formic acid plus enzyme treatments enhanced (P<0. 05) water soluble carbohydrate content significantly compared with control at all ensiling periods. The lactic acid content and acetic acid content were higher (P<0. 05) in the enzyme treatment than those in the formic acid-contained treatments and control, respectively. However, the acetic acid content was lower ( P<0. 05 ) in formic acid-contained treatments than that in enzymes treated silages. Relative to control, all treatments had lower (P< 0. 05 ) ammonia-N concentrations during ensiling. The enzyme treatments effectively (P<0. 05) decreased neutral detergent fiber and acid detergent fiber contents in the silages. The results suggested that the addition of formic acid and enzymes improved the Lolium multiflorum and Trifolium repens mixture silage quality, and the enzyme treatments were better than formic acid treatments during ensiling.%为评价甲酸与纤维素酶和木聚糖酶处理后多花黑麦草与白三叶混合青贮料发酵品质的变化,试验将混合青贮料分为对照组(未处理)、甲酸添加组、纤维素酶添加组、甲酸+纤维素酶添加组、木聚糖酶添加组、甲酸+木聚糖酶添加组、纤维素酶+木聚糖酶添加组、甲酸+纤维素酶+木聚糖酶添加组共8组,每组3个重复.青贮原料按80

  3. Adaptation of marine derived fungus Chaetomium globosum (NIOCC 36) to alkaline stress using antioxidant properties

    Digital Repository Service at National Institute of Oceanography (India)

    Ravindran, C.; Naveenan, T.

    components. 4 2. Materials and methods 2.1. Chemicals Sodium bicarbonate, Folins–Ciocalteu reagent, Gallic acid, Sodium nitrite (NaNO 2 ), Aluminium chloride (AlCl 3 ), Sodium hydroxide (NaOH), Quercetin, 1, Ethanol, xylenol orange, Butylated... modifications. The extract (100µl) was mixed with 300 µl distilled water and 30µl of 5% NaNO 2 . After 5 min 30 µl of 10% AlCl 3 was added and mixed well. The mixture was incubated for 5 min and 0.2 ml of 1 mM NaOH was added. Finally, the volume was made...

  4. Chaetomium-like fungi causing opportunistic infections in humans: a possible role for extremotolerance

    NARCIS (Netherlands)

    Ahmed, Sarah A.; Khan, Ziauddin; Wang, Xue-wei; Moussa, Tarek A. A.; Al-Zahrani, Hassan S.; Almaghrabi, Omar A.; Sutton, Deanna A.; Ahmad, S.; Groenewald, Johannes Z.; Alastruey-Izquierdo, A.; Diepeningen, Anne; Menken, S. B. J.; Najafzadeh, M. J.; Crous, Pedro W.; Cornely, Oliver; Hamprecht, Axel; Vehreschild, Maria J. G. T.; Kindo, A. J.; de Hoog, G. Sybren

    2016-01-01

    Members of the family Chaetomiaceae are ubiquitous ascosporulating fungi commonly, which reside in soil enriched with manure or cellulosic materials. Their role as human pathogens is largely ignored. However, the ability of some species to grow at high temperature enables them to play an important r

  5. Chaetomium-like fungi causing opportunistic infections in humans: a possible role for extremotolerance

    NARCIS (Netherlands)

    S.A. Ahmed; Z. Khan; X. Wang; T.A.A. Moussa; H.S. Al-Zahrani; O.A. Almaghrabi; D.A. Sutton; S. Ahmad; J.Z. Groenewald; A. Alastruey-Izquierdo; A. van Diepeningen; S.B.J. Menken; M.J. Najafzadeh; P.W. Crous; O. Cornely; A. Hamprecht; M.J.G.T. Vehreschild; A.J. Kindo; G.S. de Hoog

    2015-01-01

    Members of the family Chaetomiaceae are ubiquitous ascosporulating fungi commonly, which reside in soil enriched with manure or cellulosic materials. Their role as human pathogens is largely ignored. However, the ability of some species to grow at high temperature enables them to play an important r

  6. Influence of soil saprophyte fungus Chaetomium cochliodes on associative system "Triticum aestivum – Azospirillum brasilense"

    OpenAIRE

    E. P. Kopylov; A. A. Zhidenko

    2009-01-01

    In laboratory and vegetative experiments the ability of soil ascomycete C. cochliodes 3250 to promote the penetration of Azospirillum nitrogen-fixing bacteria into roots’ inner tissues was shown. At the same time the endophytic association: spring wheat – Azospirillum nitrogen-fixing bacteria – soil saprophyte ascomycete C. cochliodes 3250 is forming. It allows activating the nitrogen fixation in the spring wheat roots zone and biosynthetic processes in plants, in particular: to raise glutami...

  7. Scientific Opinion on the safety and efficacy of Endofeed® DC (endo-1,3(4-beta-glucanase and endo-1,4-beta-xylanase as a feed additive for chickens for fattening, laying hens, pigs for fattening and minor poultry and porcine species

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2013-08-01

    Full Text Available The additive Endofeed® DC contains endo-1,3(4-beta-glucanase and endo-1,4-beta-xylanase from the fermentation product produced by a non-genetically modified strain of Aspergillus niger. The applicant seeks the re-evaluation of the product and the extension of its use to pigs for fattening, minor poultry and porcine species. Tolerance trials in chickens for fattening, laying hens and pigs showed that the animals tolerated well 100-fold the recommended dose. Therefore, the FEEDAP Panel concludes that the additive is safe for these target species. The conclusions reached on those species can be extrapolated to minor poultry species and to minor porcine species for fattening. In the absence of adequate studies, the FEEDAP Panel cannot conclude on the safety of Endofeed® DC for the consumer. No specific studies were provided regarding the safety for the user. Therefore, Endofeed® DC should be considered a potential skin and eye irritant, and a potential skin and respiratory sensitiser. The results obtained in efficacy studies performed in chickens for fattening, laying hens and pigs for fattening showed that the additive has the potential to improve the performance of the animals. The nominal dose at which the potential of the enzyme preparation is demonstrated is the recommended dose. The conclusions on the efficacy on the major poultry species and on pigs for fattening can be extrapolated to minor poultry species and minor porcine species for fattening, respectively.

  8. Scientific Opinion on the safety and efficacy of Rovabio® Spiky (endo-1,4-beta-xylanase and endo-1,3(4-beta-glucanase as a feed additive for chickens for fattening, chickens reared for laying and other minor poultry species (for fattening and reared for laying

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2014-07-01

    Full Text Available Rovabio® Spiky is an enzyme preparation, available in solid and liquid forms, of endo-1,4-beta-xylanase and endo-1,3(4-beta-glucanase. The enzymes present in the additive are produced by two strains of Penicillium funiculosum, one of them genetically modified. The additive is intended to be used as a feed additive for chickens for fattening, chickens reared for laying and other minor poultry species (for fattening and reared for laying. None of the production strains was detected in their respective products. The additive does not give rise to safety concerns with regard to the genetic modification. No recombinant DNA was detected in the product obtained from the genetically modified strain of P. funiculosum. Based on the results of a tolerance trial in chickens for fattening, the FEEDAP Panel concludes that the additive is safe for chickens for fattening under the recommended conditions of use. This conclusion can be extended to chickens reared for laying and can be extrapolated to minor poultry species for fattening or reared for laying. Based on the outcome of the toxicological studies performed with the products of fermentation used to formulate the additive, the additive is of no concern regarding consumer safety. The additive is not irritant to the skin or eyes. In the absence of data, it should be considered a potential skin sensitiser and potentially harmful if inhaled. No risks to the environment are expected from the use of the additive in animal nutrition. Based on the results obtained in three efficacy studies, the FEEDAP Panel concludes that the additive has the potential to be efficacious in chickens for fattening at the minimum recommended dose. This conclusion can be extended to chickens reared for laying and can be extrapolated to minor poultry species for fattening or reared for laying.

  9. Optimization of alkaline cellulase production by the marine-derived fungus Chaetomium sp. using agricultural and industrial wastes as substrates

    Digital Repository Service at National Institute of Oceanography (India)

    Ravindran, C.; Naveenan, T.; Varatharajan, G.R.

    , waste paper) with or without mineral salt solution (gL -1 , w/v): NaNO 3 , 3.0; KH 2 PO 4 , 1.0; Tween 80, 1.0 mL; MgSO 4 .7H 2 O, 0.5; KCl. 0.5; FeSO 4 . 7H 2 O, 0.01 was added to the 500 ml flask, moistened with ~ 20 ml of water. The flasks were...

  10. 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.

  11. In TFIIH, XPD helicase is exclusively devoted to DNA repair.

    Directory of Open Access Journals (Sweden)

    Jochen Kuper

    2014-09-01

    Full Text Available The eukaryotic XPD helicase is an essential subunit of TFIIH involved in both transcription and nucleotide excision repair (NER. Mutations in human XPD are associated with several inherited diseases such as xeroderma pigmentosum, Cockayne syndrome, and trichothiodystrophy. We performed a comparative analysis of XPD from Homo sapiens and Chaetomium thermophilum (a closely related thermostable fungal orthologue to decipher the different molecular prerequisites necessary for either transcription or DNA repair. In vitro and in vivo assays demonstrate that mutations in the 4Fe4S cluster domain of XPD abrogate the NER function of TFIIH and do not affect its transcriptional activity. We show that the p44-dependent activation of XPD is promoted by the stimulation of its ATPase activity. Furthermore, we clearly demonstrate that XPD requires DNA binding, ATPase, and helicase activity to function in NER. In contrast, these enzymatic properties are dispensable for transcription initiation. XPD helicase is thus exclusively devoted to NER and merely acts as a structural scaffold to maintain TFIIH integrity during transcription.

  12. Using Group II Introns for Attenuating the In Vitro and In Vivo Expression of a Homing Endonuclease.

    Directory of Open Access Journals (Sweden)

    Tuhin Kumar Guha

    Full Text Available In Chaetomium thermophilum (DSM 1495 within the mitochondrial DNA (mtDNA small ribosomal subunit (rns gene a group IIA1 intron interrupts an open reading frame (ORF encoded within a group I intron (mS1247. This arrangement offers the opportunity to examine if the nested group II intron could be utilized as a regulatory element for the expression of the homing endonuclease (HEase. Constructs were generated where the codon-optimized ORF was interrupted with either the native group IIA1 intron or a group IIB type intron. This study showed that the expression of the HEase (in vivo in Escherichia coli can be regulated by manipulating the splicing efficiency of the HEase ORF-embedded group II introns. Exogenous magnesium chloride (MgCl2 stimulated the expression of a functional HEase but the addition of cobalt chloride (CoCl2 to growth media antagonized the expression of HEase activity. Ultimately the ability to attenuate HEase activity might be useful in precision genome engineering, minimizing off target activities, or where pathways have to be altered during a specific growth phase.

  13. Bifunctional xylanases and their potential use in biotechnology

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Numan, M.Th.

    of glucan units in the polymer, reVered as the degree of polymeriza- tion. The degree of polymerization of cellulose depends on the type of plants and typically is estimated to be from 2,000 to 27,000 glucan units [60]. Hemicellulose belongs to a group... activity, and C-terminal domain coding for putative polysaccharide deacylase implicated in removing acetyl groups from acetylated xylan, and thus it is probably capa- ble of hydrolyzing acetylated xylan debranching in the xylan backbone [14]. Cytophaga...

  14. Heterologous Expression of Xylanase Enzymes in Lipogenic Yeast Yarrowia lipolytica

    OpenAIRE

    Wang, Wei; Wei, Hui; Alahuhta, Markus; Chen, Xiaowen; Hyman, Deborah; Johnson, David K; Zhang, Min; Himmel, Michael E.

    2014-01-01

    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 ...

  15. 嗜极性木聚糖酶%Extremophilic Xylanase

    Institute of Scientific and Technical Information of China (English)

    郑春翠; 段文凯; 周晓云

    2007-01-01

    木聚糖酶内切水解木聚糖主链的1,4-β-D-糖苷键,木聚糖是植物细胞壁中一种主要的多糖.自然界中木聚糖是多种糖类的复合体,这就使得木聚糖酶呈现多态性和多域性,由此需将繁多的木聚糖酶进行归类.木聚糖酶的催化反应属于双置换机制.在已研究的真菌或细菌性木聚糖酶中,大多数在温和的条件下表现出最佳活性,但有很多在极端环境下生长的生物体,为了适应极端环境而产生嗜极性的酶,其中嗜酸的、嗜碱的、嗜热的木聚糖酶,现在已有广泛的研究.对嗜极性木聚糖酶的研究进展作了论述.

  16. Bioabatement with xylanase supplementation to reduce enzymatic hydrolysis inhibitors

    Science.gov (United States)

    Bioabatement, using the fungus Coniochaeta ligniaria NRRL30616 can effectively eliminate enzyme inhibitors from pretreated biomass hydrolysis. However, our recent research suggested that bioabatement had no beneficial effect on removing xylo-oligomers which were identified as strong inhibitors to ce...

  17. Symbiodinium thermophilum sp. nov., a thermotolerant symbiotic alga prevalent in corals of the world's hottest sea, the Persian/Arabian Gulf

    OpenAIRE

    Hume, B. C. C.; .C. D'Angelo; Smith, E. G.; Stevens, J.R.; Burt, J.; Wiedenmann, J.

    2015-01-01

    Coral reefs are in rapid decline on a global scale due to human activities and a changing climate. Shallow water reefs depend on the obligatory symbiosis between the habitat forming coral host and its algal symbiont from the genus Symbiodinium (zooxanthellae). This association is highly sensitive to thermal perturbations and temperatures as little as 1°C above the average summer maxima can cause the breakdown of this symbiosis, termed coral bleaching. Predicting the capacity of corals to surv...

  18. Endophytic fungi associated with endogenous Boswellia sacra

    Directory of Open Access Journals (Sweden)

    SAIFELDIN A.F. EL-NAGERABI

    2014-04-01

    Full Text Available El-Nagerabi SAF, Elshafie AE, AlKhanjari SS. 2014. Endophytic fungi associated with endogenous Boswellia sacra. Biodiversitas 15: 22-28. Endophytic fungi associated with leaves and stem tissues of Boswellia sacra growing in Dhofar Mountains of Oman were investigated from May 2008 through October 2011. The biological diversity, tissue-preference and seasonal variations of fungi were evaluated. Forty-three species and 3 varieties of fungi were recovered as new records from this plant. Of these isolates, 35 species are new reports to the mycoflora of Oman, whereas 12 species were added to the list of fungal flora of the Arabian Peninsula. The genus Alternaria (12 species is the most prevalent genus recovered from 12.5-83.3% of the screened leaves and stem samples, followed by Aspergillus (5 species, 3 varieties, 6.9-86.1%, Mycelia sterilia (76.4%, Rhizopus stolonifer (62.5%, Drechslera (3 species, 40.3-54.2%, Cladosporium (3 species, 20.8-52.8%, Curvularia lunata (38.8%, Chaetomium (2 species, 15.3-26.3%, Penicillim spp. (9.8-27.8%, Fusarium (9 species, 6.9-27.8%, Ulocladium consortiale (27.8%, Mucor hiemalis (19.5%, and the remaining species (Scytalidium thermophilum, Phoma solani, Taeniolella exilis, and Botryodiplodia theobromae exhibited very low levels of incidence (4.2-11.1%. Endophytic colonization of the leaf tissues was greater (43 species, 3 varieties comparable to stem tissues (25 species. This indicates heterogeneity and tissue-preference, with no evidence of seasonal variation. Therefore, the isolation of many fungal species and sterile mycelia supports the biodiversity of the endophytic fungi invading B. sacra and the high possibility of isolating more fungal species using advanced molecular techniques.

  19. Architecture of the eIF2B regulatory subcomplex and its implications for the regulation of guanine nucleotide exchange on eIF2.

    Science.gov (United States)

    Kuhle, Bernhard; Eulig, Nora K; Ficner, Ralf

    2015-11-16

    Eukaryal translation initiation factor 2B (eIF2B) acts as guanine nucleotide exchange factor (GEF) for eIF2 and forms a central target for pathways regulating global protein synthesis. eIF2B consists of five non-identical subunits (α-ϵ), which assemble into a catalytic subcomplex (γ, ϵ) responsible for the GEF activity, and a regulatory subcomplex (α, β, δ) which regulates the GEF activity under stress conditions. Here, we provide new structural and functional insight into the regulatory subcomplex of eIF2B (eIF2B(RSC)). We report the crystal structures of eIF2Bβ and eIF2Bδ from Chaetomium thermophilum as well as the crystal structure of their tetrameric eIF2B(βδ)2 complex. Combined with mutational and biochemical data, we show that eIF2B(RSC) exists as a hexamer in solution, consisting of two eIF2Bβδ heterodimers and one eIF2Bα2 homodimer, which is homologous to homohexameric ribose 1,5-bisphosphate isomerases. This homology is further substantiated by the finding that eIF2Bα specifically binds AMP and GMP as ligands. Based on our data, we propose a model for eIF2B(RSC) and its interactions with eIF2 that is consistent with previous biochemical and genetic data and provides a framework to better understand eIF2B function, the molecular basis for Gcn(-), Gcd(-) and VWM/CACH mutations and the evolutionary history of the eIF2B complex. PMID:26384431

  20. Endophytic fungi associated with endogenous Boswellia sacra

    Directory of Open Access Journals (Sweden)

    SAIFELDIN A.F. EL-NAGERABI1,♥,

    2014-11-01

    Full Text Available Endophytic fungi associated with leaves and stem tissues of Boswellia sacra growing in Dhofar Mountains of Oman were investigated from May 2008 through October 2011. The biological diversity, tissue-preference and seasonal variations of fungi were evaluated. Forty-three species and 3 varieties of fungi were recovered as new records from this plant. Of these isolates, 35 species are new reports to the mycoflora of Oman, whereas 12 species were added to the list of fungal flora of the Arabian Peninsula. The genus Alternaria (12 species is the most prevalent genus recovered from 12.5-83.3% of the screened leaves and stem samples, followed by Aspergillus (5 species, 3 varieties, 6.9-86.1%, Mycelia sterilia (76.4%, Rhizopus stolonifer (62.5%, Drechslera (3 species, 40.3-54.2%, Cladosporium (3 species, 20.8-52.8%, Curvularia lunata (38.8%, Chaetomium (2 species, 15.3-26.3%, Penicillim spp. (9.8-27.8%, Fusarium (9 species, 6.9-27.8%, Ulocladium consortiale (27.8%, Mucor hiemalis (19.5%, and the remaining species (Scytalidium thermophilum, Phoma solani, Taeniolella exilis, and Botryodiplodia theobromae exhibited very low levels of incidence (4.2-11.1%. Endophytic colonization of the leaf tissues was greater (43 species, 3 varieties comparable to stem tissues (25 species. This indicates heterogeneity and tissue-preference, with no evidence of seasonal variation. Therefore, the isolation of many fungal species and sterile mycelia supports the biodiversity of the endophytic fungi invading B. sacra and the high possibility of isolating more fungal species using advanced molecular techniques.

  1. Deep transcriptome-sequencing and proteome analysis of the hydrothermal vent annelid Alvinella pompejana identifies the CvP-bias as a robust measure of eukaryotic thermostability

    Directory of Open Access Journals (Sweden)

    Holder Thomas

    2013-01-01

    Full Text Available Abstract Background Alvinella pompejana is an annelid worm that inhabits deep-sea hydrothermal vent sites in the Pacific Ocean. Living at a depth of approximately 2500 meters, these worms experience extreme environmental conditions, including high temperature and pressure as well as high levels of sulfide and heavy metals. A. pompejana is one of the most thermotolerant metazoans, making this animal a subject of great interest for studies of eukaryotic thermoadaptation. Results In order to complement existing EST resources we performed deep sequencing of the A. pompejana transcriptome. We identified several thousand novel protein-coding transcripts, nearly doubling the sequence data for this annelid. We then performed an extensive survey of previously established prokaryotic thermoadaptation measures to search for global signals of thermoadaptation in A. pompejana in comparison with mesophilic eukaryotes. In an orthologous set of 457 proteins, we found that the best indicator of thermoadaptation was the difference in frequency of charged versus polar residues (CvP-bias, which was highest in A. pompejana. CvP-bias robustly distinguished prokaryotic thermophiles from prokaryotic mesophiles, as well as the thermophilic fungus Chaetomium thermophilum from mesophilic eukaryotes. Experimental values for thermophilic proteins supported higher CvP-bias as a measure of thermal stability when compared to their mesophilic orthologs. Proteome-wide mean CvP-bias also correlated with the body temperatures of homeothermic birds and mammals. Conclusions Our work extends the transcriptome resources for A. pompejana and identifies the CvP-bias as a robust and widely applicable measure of eukaryotic thermoadaptation. Reviewer This article was reviewed by Sándor Pongor, L. Aravind and Anthony M. Poole.

  2. Compositional Changes and Baking Performance of Rye Dough As Affected by Microbial Transglutaminase and Xylanase.

    Science.gov (United States)

    Grossmann, Isabel; Döring, Clemens; Jekle, Mario; Becker, Thomas; Koehler, Peter

    2016-07-20

    Doughs supplemented with endoxylanase (XYL) and varying amounts of microbial transglutaminase (TG) were analyzed by sequential protein extraction, quantitation of protein fractions and protein types, and determination of water-extractable arabinoxylans. With increasing TG activity, the concentration of prolamins and glutelins decreased and increased, respectively, and the prolamin-to-glutelin ratio strongly declined. The overall amount of extractable protein decreased with increasing TG level showing that cross-linking by TG provided high-molecular-weight protein aggregates. The decrease of the high-molecular-weight arabinoxylan fraction and the concurrent increase of the medium-molecular-weight fraction confirmed the degradation of arabinoxylans by XYL. However, XYL addition did not lead to significant improved cross-linking of rye proteins by TG. Volume and crumb hardness measurements of bread showed increased protein connectivity induced by XYL and TG. Significant positive effects on the final bread quality were especially obtained by XYL addition. PMID:27349134

  3. Shear-induced starch-gluten separation at very low water content aided by xylanases

    NARCIS (Netherlands)

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

    2014-01-01

    This study examines the influence of extremely low water content on shear-induced starch–gluten separation and how endoxylanases influence the separation by releasing water associated with arabinoxylan. Shearing was performed at a water content ranging from 34% to 43.5% (w/w). It was possible to con

  4. Studies on substrate specificity of β-xylanase from Streptomyces olivaceoviridis E-86

    OpenAIRE

    Yoshida, Shigeki

    1996-01-01

    Plant cell wall is the major reservoir of fixed carbon in nature,and consists of three major polymeric components,namely cellulose,hemicellulose,and lignin.β-1,4-Xylans are the most aboundant components of the hemicellulose and mainly found ...

  5. Application of thermoalkalophilic xylanase from Arthrobacter sp. MTCC 5214 in biobleaching of kraft pulp

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Bhosle, N.B.

    released by enzyme treatment showed a characteristic peak at 280 nm indicating the presence of lignin in the released coloring matter. Enzymatic prebleaching of kraft pulp showed 20 % reduction in kappa number of the pulp without much change in viscosity...

  6. Improvement in the productivity of xylooligosaccharides from rice straw by feed xylanase with ultrafiltration

    OpenAIRE

    Wang Fei; Guohua Hu; Xiao Jianbo; Liu Yang

    2011-01-01

    The effective production of xylooligosaccharides (XOs) from rice straw was investigated. Rice straw contains rich hemicellulose which can be hydrolyzed by enzyme; the XOs were obtained under hydrothermal conditions. To improve the productivity of XOs, ultrafiltration was chosen to eliminate xylan in the XOs. Under optimum hydrolysis conditions (1000 IU enzyme/g, 35 0C, 10% substrate concentration, pH 6.5, 6 h), the DP was the lowest. After ultrafiltration, xylan was eliminated. On the b...

  7. Improvement in the productivity of xylooligosaccharides from rice straw by feed xylanase with ultrafiltration

    Directory of Open Access Journals (Sweden)

    Wang Fei

    2011-01-01

    Full Text Available The effective production of xylooligosaccharides (XOs from rice straw was investigated. Rice straw contains rich hemicellulose which can be hydrolyzed by enzyme; the XOs were obtained under hydrothermal conditions. To improve the productivity of XOs, ultrafiltration was chosen to eliminate xylan in the XOs. Under optimum hydrolysis conditions (1000 IU enzyme/g, 35 0C, 10% substrate concentration, pH 6.5, 6 h, the DP was the lowest. After ultrafiltration, xylan was eliminated. On the basis of experimental data, an industrial XO production process consisting of pretreatment, enzymatic treatment and purification was designed. Using the designed process, 2.9g dry of purified XO was produced from 50g dry rice straw power.

  8. Statistical optimization of thermo-tolerant xylanase activity from Amazon isolated Bacillus circulans on solid-state cultivation.

    Science.gov (United States)

    Heck, Júlio Xandro; Flôres, Simone Hickmann; Hertz, Plinho Francisco; Ayub, Marco Antônio Záchia

    2006-10-01

    A 2(2) factorial design was performed to find the best conditions of pH and temperature for xylanolytic activity of Bacillus circulans BL53 isolated from the Amazon environment. Solid-state cultivation was carried out on an inexpensive, abundant agro-industrial soybean residue. The central composite design (CCD) used for the analysis of treatment combinations showed that a second-order polynomial regression model was in good agreement with experimental results, with R(2) = 0.9369 (P processes. The crude enzyme extract hydrolyzed rice straw, sugar cane bagasse and soybean fiber and its activity was stimulated by Co(2+), Fe(3+), and beta-mercaptoethanol but inhibited by Mn(2+), Cu(2+), Ca(2+), Zn(2+), Ba(2+), Mg(2+) and by EDTA. PMID:16216495

  9. The genome of the mustard leaf beetle encodes two active xylanases originally acquired from bacteria through horizontal gene transfer

    OpenAIRE

    Pauchet, Yannick; Heckel, David G

    2013-01-01

    The primary plant cell wall comprises the most abundant polysaccharides on the Earth and represents a rich source of energy for organisms which have evolved the ability to digest them. Enzymes able to degrade plant cell wall polysaccharides are widely distributed in micro-organisms but are generally absent in animals, although their presence in insects, especially phytophagous beetles from the superfamilies Chrysomeloidea and Curculionoidea, has recently begun to be appreciated. The observed ...

  10. Partial Optimization of Endo-1, 4-Β-Xylanase Production by Aureobasidium pullulansUsing Agro-Industrial Residues

    Directory of Open Access Journals (Sweden)

    Shaghayegh Nasr

    2013-12-01

    This finding indicates the feasibility of growing of A. pullulans strain SN090 on wheat bran as an alternate economical substrate in order for reducing the costs of enzyme production and using this fortified agro-industrial byproduct in formulation of animal feed.

  11. Cellulose and hemicellulose-degrading enzymes in Fusarium commune transcriptome and functional characterization of three identified xylanases

    DEFF Research Database (Denmark)

    Yuhong, Huang; Busk, Peter Kamp; Lange, Lene

    2015-01-01

    Specific enzymes from plant-pathogenic microbes demonstrate high effectiveness for natural lignocellulosic biomass degradation and utilization. The secreted lignocellulolytic enzymes of Fusarium species have not been investigated comprehensively, however. In this study we compared cellulose...... and hemicellulose-degrading enzymes of classical fungal enzyme producers with those of Fusarium species. The results indicated that Fusarium species are robust cellulose and hemicellulose degraders. Wheat bran, carboxymethylcellulose and xylan-based growth media induced a broad spectrum of lignocellulolytic enzymes...... in Fusarium commune. Prediction of the cellulose and hemicellulose-degrading enzymes in the F. commune transcriptome using peptide pattern recognition revealed 147 genes encoding glycoside hydrolases and six genes encoding lytic polysaccharide monooxygenases (AA9 and AA11), including all relevant cellulose...

  12. fRNAdb Summary: FR068821 [

    Lifescience Database Archive (English)

    Full Text Available FR068821 AB019364,AB019365,AB038564,AB038565,AB038568,AB038569,AB038570,AB038573,AB038574,AB0385 ... s,Paecilomyces penicillatus,Chaetomium sp. 73-19-O-Mexico ,foliar endophyte of Picea glauca sp. O6,Chaetomium ...

  13. Growth-promoting effect of thermophilic fungi on the mycelium of the edible mushroom Agaricus bisporus.

    OpenAIRE

    Wiegant, W.M.; Wery, J.; E. T. Buitenhuis; de Bont, J A

    1992-01-01

    The growth-promoting effect of the thermophilic fungus Scytalidium thermophilum in mushroom compost on the mycelium of the edible mushroom Agaricus bisporus was investigated. Results obtained by others were confirmed by showing that S. thermophilum leads to an increased hyphal extension rate of the mushroom mycelium. However, it was demonstrated that hyphal extension rates were not clearly related to mushroom biomass increase rates. A number of experiments pointed strongly towards CO2 as the ...

  14. BLEACHING OF SULFONATED CMP FROM BIO-TREATED WHEAT STRAW

    Institute of Scientific and Technical Information of China (English)

    HongYu; MenghuaQin; XuemeiLu; YinboQu; PeijiGao

    2004-01-01

    Wheat straw chemi-mechanical pulp was pretreated with a crude xylanase which was secreted by white rot fungus Phanerochaete Chrysosporium prior to hydrogen peroxide bleaching. The process of xylanase pretreatment and hydrogen peroxide bleaching was optimized. The xylanase treated pulp achieved a brightness gain of 5.8% ISO over the untreated pulp. The xylanase treatment was found to liberate reducing sugars and facilitating lignin removal. Fiber morphology of pulp treated with xylanase was also studied by SEM.

  15. Enzymatic hydrolysis of water-soluble wheat arabinoxylan. 1. Synergy between alpha-L-arabinofuranosidases, endo-1,4-beta-xylanases, and beta-xylosidase activities

    DEFF Research Database (Denmark)

    Sørensen, H.R.; Meyer, Anne Boye Strunge; Pedersen, S.

    2003-01-01

    Hydrolysis of arabinoxylan is an important prerequisite for improved utilization of wheat hemicellulose in the ethanol fermentation industry. This study investigates the individual and combined efficiencies of three commercial, cellulytic and hemicellulytic enzyme preparations, Celluclast 1.5 L, ...

  16. Purification of Clostridium thermocellum xylanase Z expressed in Escherichia coli and identification of the corresponding product in the culture medium of C. thermocellum.

    OpenAIRE

    Grépinet, O.; Chebrou, M C; Béguin, P

    1988-01-01

    An endoxylanase encoded by the xynZ gene of Clostridium thermocellum was purified from Escherichia coli harbouring a fragment of the gene cloned in pUC8. The purified enzyme showed two active bands of Mr 41,000 and 39,000, the latter one presumably derived from the former through proteolysis. The enzyme was highly active on xylan and para-nitrophenyl-beta-D-xylobioside. The major end product of xylan hydrolysis was xylobiose. With an antiserum raised against the enzyme purified from E. coli, ...

  17. Effects of Exogenous NSP Enzymes(Xylanase, β-glucanase and Cellulase) on Morphology and Functions of Digestive Tract in Growing Pigs Fed with Paddy-Based Diets

    Institute of Scientific and Technical Information of China (English)

    XU Zi-rong; LU Jian-jun

    2003-01-01

    Ninety Landrace X Jia 35±0.40 kg weight growing pigs were randomly allotted to three treatments, each of which was replicated three times with ten pigs per replicate. The pigs were reared on either a conventional corn-based diet (control Ⅰ ) or a paddy-based diet (control Ⅱ ) or a paddy diet supplemented with 0.2% NSP enzymes (test group). All pigs were given ad libitum access to both feed and water. The results of feeding trial showed that supplementation of NSP enzymes significantly increased ADG by 8.78 % (P<0.05) and decreased F/G by 9.42% (P<0.05) over the control group Ⅱ. No significant differences were found in ADG and F/G between control group I and the test group. The digestive trial showed that adding NSP enzymes significantly improved apparent digestibility of CP, EE and CF by 18.76 (P<0.01), 16.04 (P <0.05) and 108.57%(P<0.05), respectively, compared to control Ⅱ. The activities of proteolytic enzyme and α-amylase in duodenal contents were increased by 99.07 (P<0.01) and 18.41% (P<0. 05) with the addition of NSP enzymes. No significant differences between test and control Ⅱ group were found in activities of the pepsin in the gastric content, the trypsin and lipase in duodenal contents, the disaccharidase and γ-glutany transferase (γ-GT) in intestinal mucosa, but there was a tendency towards higher activities associated with the NSP enzymes diet (P>0. 05). The lengths of the villi within the duodenal, jejunal and ileal sections of the small intestine of pigs receiving the NSP enzymes diet were increased by 23.68 (P<0. 05), 56.00 (P<0. 01)and 76.90%(P<0.01) respectively, relative to the pigs in control Ⅱ.

  18. Construction of a Xylanase-Producing Strain of Brevibacterium lactofermentum by Stable Integration of an Engineered xysA Gene from Streptomyces halstedii JM8

    OpenAIRE

    Adham, Sirin A.I.; Campelo, Ana B.; Ramos, Angelina; Gil, José A.

    2001-01-01

    A xylanolytic strain of Brevibacterium lactofermentum containing the Streptomyces halstedii His-tagged xysA gene was generated. The new strain contains DNA derived from S. halstedii, expresses xylanolytic activity, and was obtained by an integrative process mediated by a conjugative plasmid targeted to a dispensable chromosomal region located downstream from the essential cell division gene ftsZ. The His-tagged Xys1 enzyme was constitutively expressed under the control of the kan promoter fro...

  19. 交联木聚糖酶聚集体的制备及其性质研究%Preparation and Characteristics of Cross-linked Xylanase Aggregates

    Institute of Scientific and Technical Information of China (English)

    任延刚; 朱启忠; 黄庆瑞; 王娜; 张瑞景

    2009-01-01

    制备了交联木聚糖酶聚集体(CLEAs),并对其酶学性质进行了研究,以提高木聚糖酶的稳定性.通过毛栓菌发酵分离得木聚糖酶,经硫酸铵沉淀后以戊二醛作为交联剂对其进行化学交联,制得CLEAs,并用常规方法测定了各种因素对游离木聚糖酶和CLEAs的影响.在对耐热性、有机相中稳定性和耐酸碱性的研究中,CLEAs均表现比游离酶有较高的稳定性,因此运用CLEA技术可提高木聚糖酶的稳定性,使其广泛应用于饲料等工业生产中.

  20. 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

    polysaccharide glucose by 21% compared with the control (P = 0.005). Compared with the control, addition of EX, EXP, and CAX decreased the concentration of soluble arabinoxylan in ileal digesta by 40 (P ... increased the concentration of LMW arabinoxylan in ileal digesta by 40 (P = 0.0001), 36 (P = 0.0006), and 24% (P = 0.023), respectively, compared with the control. Addition of EX and EXP decreased the concentration of soluble NSP of ileal digesta by 25 (P = 0.001) and 26% (P

  1. Draft Genome Sequence of Pseudozyma brasiliensis sp. nov. Strain GHG001, a High Producer of Endo-1,4-Xylanase Isolated from an Insect Pest of Sugarcane.

    Science.gov (United States)

    Oliveira, Juliana Velasco de Castro; Dos Santos, Renato Augusto Corrêa; Borges, Thuanny A; Riaño-Pachón, Diego Mauricio; Goldman, Gustavo Henrique

    2013-01-01

    Here, we present the nuclear and mitochondrial genome sequences of Pseudozyma brasiliensis sp. nov. strain GHG001. P. brasiliensis sp. nov. is the closest relative of Pseudozyma vetiver. P. brasiliensis sp. nov. is capable of growing on xylose or xylan as a sole carbon source and has great biotechnological potential. PMID:24356824

  2. Effect of heat-treatment, phytase, xylanase and soaking time on inositol phosphate degradation in vitro in wheat, soybean meal and rapeseed cake

    DEFF Research Database (Denmark)

    Blaabjerg, Karoline; Carlsson, N G; Hansen-Møller, Jens;

    2010-01-01

    An in vitro method was used to evaluate the degradation of myo-inositol hexakisphosphate (InsP6) in non-heat-treated wheat (NHW), heat-treated wheat (HW), soybean meal (SBM) or rapeseed cake (RSC) soaked separately or in combination. The feedstuffs were soaked in water (20 °C) and samples were...

  3. Laboratory research on the efficacy of chlorine dioxide fumigation for the remediation of mold-contaminated buildings--conference paper

    Science.gov (United States)

    The purpose of this project was to determine the efficacy ofCl02 fumigation to inactivate viable mold, mycotoxins, and allergens on building materials. Alternaria alternata, Aspergillus versicolor, Aspergillus Jumigatus, Chaetomium globosum, and Stachybotrys chartarum were indivi...

  4. Purification, biochemical characterization and structural modelling of alkali-stable β-1,4-xylan xylanohydrolase from Aspergillus fumigatus R1 isolated from soil

    OpenAIRE

    Deshmukh, Rehan Ahmed; Jagtap, Sharmili; Mandal, Madan Kumar; Mandal, Suraj Kumar

    2016-01-01

    Background Aspergillus fumigatus R1 produced xylanase under submerged fermentation which degrades the complex hemicelluloses contained in agricultural substrates. Xylanases have gained considerable attention because of their tremendous applications in industries. The purpose of our study was to purify xylanase and study its biochemical properties. We have predicted the secondary structure of purified xylanase and evaluated its active site residues and substrate binding sites based on the glob...

  5. Gene : CBRC-XTRO-01-0641 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-0641 Novel UN C UNKNOWN ALSC_ECOLI 1e-106 60% ref|YP_076170.1| sugar ... ABC transporte ... obacterium thermophilum IAM 14863] dbj|BAD41326.1| sugar ... ABC transporter ATP-binding protein [Symbiobacteri ...

  6. Separation and identification of endoxylanases from Bacillus subtilis and their actions on wheat bran insoluble dietary fibre

    OpenAIRE

    Xiaoping, Yuan; Jing, Wang; Huiyuan, Yao; Nihorimbere, Venant

    2005-01-01

    A novel and convenient method based on native polyacrylamide gel electrophoresis (PAGE) and homogenization extraction was used for the purification of xylanase from crude enzymes. Two xylanases were purified by this method from the crude enzyme preparation from the selected strain of Bacillus subtilis. Subsequent analysis with thin layer chromatography and high pressure liquid chromatography (HPLC) indicated that these two xylanases were endo-acting enzymes, designated xyl I and xyl II. Both ...

  7. Production, Purification, and Characterization of β-(1-4)-Endoxylanase of Streptomyces roseiscleroticus

    OpenAIRE

    Grabski, Anthony C.; Jeffries, Thomas W.

    1991-01-01

    Twelve species of Streptomyces that formerly belonged to the genus Chainia were screened for the production of xylanase and cellulase. One species, Streptomyces roseiscleroticus (Chainia rosea) NRRL B-11019, produced up to 16.2 IU of xylanase per ml in 48 h. A xylanase from S. roseiscleroticus was purified and characterized. The enzyme was a debranching β-(1-4)-endoxylanase showing high activity on xylan but essentially no activity against acid-swollen (Walseth) cellulose. It had a very low a...

  8. Purification and Properties of a Xylan-Binding Endoxylanase from Alkaliphilic Bacillus sp. Strain K-1

    OpenAIRE

    Ratanakhanokchai, Khanok; Kyu, Khin Lay; Tanticharoen, Morakot

    1999-01-01

    An alkaliphilic bacterium, Bacillus sp. strain K-1, produces extracellular xylanolytic enzymes such as xylanases, β-xylosidase, arabinofuranosidase, and acetyl esterase when grown in xylan medium. One of the extracellular xylanases that is stable in an alkaline state was purified to homogeneity by affinity adsorption-desorption on insoluble xylan. The enzyme bound to insoluble xylan but not to crystalline cellulose. The molecular mass of the purified xylan-binding xylanase was estimated to be...

  9. ВЛИЯНИЕ ПОЧВЕННОГО САПРОФИТНОГО ГРИБА CHAETOMIUM COCHLIODES НА АССОЦИАТИВНУЮ СИСТЕМУ « TRITICUM AESTIVUM – AZOSPIRILLUM BRASILENSE»

    OpenAIRE

    Копылов, Е.; Жиденко, А.

    2009-01-01

    В условиях лабораторных и вегетационных опытов показана способность почвенного сумчатого гриба C. cochliodes 3250 способствовать проникновению азотфиксирующих бактерий рода Azospirillum во внутренние ткани корней. При этом формируется эндофитная ассоциация: яровая пшеница – диазотрофы рода Azospirillum – почвенный сапрофитный гриб C. cochliodes 3250, что позволяет активизировать азотфиксацию в корневой зоне яровой пшеницы и биосинтетические процессы в растениях: повысить глутаминсинтетазную а...

  10. Microwave Influence in Fungi a Preliminary Study

    International Nuclear Information System (INIS)

    The behavior of two cellulolytic fungus species under the influence of low intensity microwaves was studied: Chaetomium globosum and Alternaria alternata. Enzyme activity of dehydrogenase complex was investigated by spectrophotometric method in order to real the effect of relatively short daily exposure times. Inhibitory effect was noticed for malate dehydrogenase and succinate dehydrogenase in both fungi while differentiated influence was revealed in alpha ceto glutarate dehydrogenase (inhibitory in Chaetomium globosum but stimulatory in Alternaria alternata). Isocitrate dehydrogenase activity was significantly stimulated in both fungi for 3 hours exposure time. (Author) 15 refs

  11. Microwave Influence in Fungi a Preliminary Study

    Energy Technology Data Exchange (ETDEWEB)

    Manoliu, A. I.; Tufescu, F. M.; Oprica, L.; Olteanu, Z.; Creanga, D. E.

    2004-07-01

    The behavior of two cellulolytic fungus species under the influence of low intensity microwaves was studied: Chaetomium globosum and Alternaria alternata. Enzyme activity of dehydrogenase complex was investigated by spectrophotometric method in order to real the effect of relatively short daily exposure times. Inhibitory effect was noticed for malate dehydrogenase and succinate dehydrogenase in both fungi while differentiated influence was revealed in alpha ceto glutarate dehydrogenase (inhibitory in Chaetomium globosum but stimulatory in Alternaria alternata). Isocitrate dehydrogenase activity was significantly stimulated in both fungi for 3 hours exposure time. (Author) 15 refs.

  12. The occurrence and distribution of cellulolytic fungi and Fusarium in seven Montagu’s Harrier (Circus pygargus

    Directory of Open Access Journals (Sweden)

    Teresa Korniłłowicz-Kowalska

    2013-12-01

    Full Text Available A total of 45 species of cellulolytic fungi and ten Fusarium species were identified. Three genera (Chaetomium, Trichoderma, Fusarium represented 80% of the frequency of cellulolytic fungi. Of them, Chaetomium globosum, Trichoderma viride and T. koningii were some of the most frequent species. A high differentiation of the richness and frequency of species of cellulolytic fungi depending on the nest and its individual layers was observed. Reasons for the differences in the frequency and species composition of the fungi were discussed.

  13. Coexpression and Application of Thermostable Xylanase and Glucuronidase%耐热木聚糖酶和葡萄糖醛酸苷酶的共表达及应用

    Institute of Scientific and Technical Information of China (English)

    沈艺红; 薛业敏; 侯静静; 许家兴; 李相前

    2015-01-01

    目的:提高海栖热袍菌(Thermotoga maritima MSB8)来源的木聚糖酶XynB和α-葡萄糖醛酸苷酶AguA生产效率并降低生产成本.方法:利用基因重组技术将海栖热袍菌的XynB和AguA基因置于不同表达盒下构建共表达载体pET-20b-xynB-aguA和pET-28a-xynB-aguA,分别转化大肠杆菌Escherichia coli JM109 (DE3)进行诱导表达,获得双酶混合物进而水解桦木木聚糖及玉米芯.结果:重组菌E.coli JM109 (DE3) /pET-28a-xynB-aguA比E.coli JM109 (DE3) /pET-20b-xynB-aguA产酶更具优势,在LB培养基中诱导培养8h,XynB和AguA的产量分别达到7.6 U/mL和0.5 U/mL,在TB培养基中培养,XynB可达到10.27 U/mL,AguA为1.5 U/mL.在80℃水解条件下,双酶比单一木聚糖酶能够更彻底降解桦木木聚糖,所得酶解液中木二糖的含量和纯度更高;从还原糖释放量及电子显微镜观察可以看出双酶液对农副产品玉米芯具有良好的降解作用.结论:XynB和AguA基因(T.maritima)的克隆共表达具有可行性,并且在生物转化、食品工业和饲料生产等领域具有潜在应用前景.

  14. The modular xylanase Xyn10A from Rhodothermus marinus is cell-attached, and its C-terminal domain has several putative homologues among cell-attached proteins within the phylum Bacteroidetes

    DEFF Research Database (Denmark)

    Karlsson, Eva Nordberg; Hachem, Maher Abou; Ramchuran, Santosh;

    2004-01-01

    the light of this, a revision of experimental data present on both Xyn10A and Man26A was performed, and the results all indicate a cell-anchoring role of the domain, suggesting that this domain represents a novel type of module that mediates cell attachment in proteins originating from members of the...

  15. DIFFERENT APPROACHES OF USING MULTIPLE CULTURE ISOLATES- FUSARIUM OXYSPORUM F8, PENICILLIUM NOTATUM 101 AND ASPERGILLIUS NIGER F7 FOR HIGHER PRODUCTION OF CELLULASE AND XYLANASE FROM PINUS ROXBURHII NEEDLES

    OpenAIRE

    DIVYA TANDON; NIVEDITA SHARMA; RICHA KAUSHAL

    2013-01-01

    Diminishing fossil fuel reserve and increasing cost of fossil products have rekindled effort on conversion of lignocelluloses to renewable fuel. Among various lignocelluloses' biomass, Pinus roxburghii is a predominant forest species which is scattered throughout the world. The continous shedding of pine needles and its exceptionally slow biodegradation in nature leads to huge accumulation of needles on earth by posing a serious threat to our environment which affects th...

  16. 基于土壤宏基因组文库筛选非培养木聚糖酶基因%Screening of xylanase genes from the soil metagenomic library based on uncultured method

    Institute of Scientific and Technical Information of China (English)

    熊科; 高乐; 杨玉焕; 崔晓亭; 李秀婷

    2015-01-01

    采用物理化学法提取土壤基因组DNA,将土壤DNA纯化后采用序列筛选策略获得非微生物纯培养方式的木聚糖酶基因.序列筛选依据木聚糖酶保守序列设计简并引物,PCR扩增获得200 bp的木聚糖酶核心序列,构建了约5 000个克隆子的序列文库.随机挑取200个克隆子测序,获得11条来自未培养微生物的木聚糖酶核心序列.并对其中细菌源序列1~ 19进行研究,利用HiTAIL-PCR扩增细菌源序列1~19的木聚糖酶基因全长,分析其编码蛋白,并将其与表达载体pET-28a(+)连接后导入E.coli表达,超声破壁后测得木聚糖酶酶活力为(19.94±0.3) U/mL.利用序列筛选获取土壤宏基因组源木聚糖酶基因的研究为筛选获得土壤未培养微生物木聚糖酶基因新资源提供了新的途径.

  17. Enzymatic Hydrolysis of Wheat Arabinoxylan by a Recombinant "Minimal" Enzyme Cocktail Containing beta-Xylosidase and Novel endo-1,4-beta-Xylanase and alpha-L-Arabinofuranosidase Activities

    DEFF Research Database (Denmark)

    Sørensen, Hanne R.; Pedersen, Sven; Jørgensen, Christel T.;

    2007-01-01

    This study describes the identification of the key enzyme activities required in a "minimal" enzyme cocktail able to catalyze hydrolysis of water-soluble and water-insoluble wheat arabinoxylan and whole vinasse, a fermentation effluent resulting from industrial ethanol manufacture from wheat. The...

  18. The preparation and properties of the immobilized Xylanase by carrier-crosslinked enzyme aggregation%固载化木聚糖酶交联酶聚集体的制备及性质

    Institute of Scientific and Technical Information of China (English)

    王冬伟; 孙谧

    2016-01-01

    目的 提高木聚糖酶YS1069的稳定性、重复利用率以及便于从反应体系中分离.方法 本文将交联酶聚集体CLEAs技术与载体固定化技术相结合,用LKZ-128氨基型树脂对木聚糖酶CLEAs进行固定化,并对其性质进行初步研究.结果及结论 采用单因素法和响应面法对影响固定化酶活性的因素进行分析和优化,获得最佳固定化条件为:pH9.0的酶液,以2 mL的异丙醇为沉淀剂,沉淀1h,以1.59 g/L的京尼平为交联剂,交联2h,20℃的环境中固定化9.25h,加酶量为8 mg时回收率最高,达到65.63%,加酶量为60 mg时酶活性最高,达到190U/g.重复使用10次后,其相对酶活仍为42.3%,说明具有良好的批次操作稳定性.该酶经过固定化后热稳定性和pH稳定性均得到了提高.

  19. Use of family 8 enzymes with xylanolytic activity in baking

    OpenAIRE

    Dutron, Agnes; Georis, Jacques; Genot, Bernard; Dauvrin, Thierry; Collins, Tony; Hoyoux, Anne; Feller, Georges

    2012-01-01

    The present invention describes a method to improve the properties of a dough and/or a baked product by adding a bread or dough-improving agent containing a enzyme with xylanolytic activity belonging to glycoside hydrolases family 8. Preferred enzymes are the psychrophilic xylanase from Pseudoalteromonas haloplanktis and the mesophilic xylanase Y from Bacillus halodurans C-125.

  20. Main: 1TE1 [RPSD[Archive

    Lifescience Database Archive (English)

    Full Text Available 1TE1 小麦 Bread Wheat Triticum aestivum Xylanase Inhibitor Protein I Precursor. Name=Xipi; Triti ... anzanares, H.J.Gilbert, N.Juge, A.Roussel The Dual Nature ... Of The Wheat Xylanase Protein Inhibitor Xip-I: Str ...

  1. Main: 1TA3 [RPSD[Archive

    Lifescience Database Archive (English)

    Full Text Available 1TA3 小麦 Bread Wheat Triticum aestivum Xylanase Inhibitor Protein I Precursor. Name=Xipi; Triti ... anzanares, H.J.Gilbert, N.Juge, A.Roussel The Dual Nature ... Of The Wheat Xylanase Protein Inhibitor Xip-I: Str ...

  2. Generation of xylooligosaccharides from microwave irradiated agroresidues using recombinant thermo-alkali-stable endoxylanase of the polyextremophilic bacterium Bacillus halodurans expressed in Pichia pastoris.

    Science.gov (United States)

    Kumar, Vikash; Satyanarayana, T

    2015-03-01

    The recombinant Pichia pastoris harboring the endoxylanase gene (TSEV1xyl) of Bacillus halodurans TSEV1 yielded a high titer of extracellular xylanase (502±23 U ml(-1)) on induction with methanol. The purified recombinant xylanase (TSEV1xyl) displayed optimal activity at 80°C and pH 9.0. The glycosylated recombinant xylanase exhibited higher thermostability (T1/2 of 45 min at 80°C) than the native enzyme (T1/2 of 35 min at 80°C). The agroresidues subjected to pretreatment (soaking in alkali followed by microwave irradiation) liberated xylooligosaccharides (XOS) upon hydrolysis with the recombinant xylanase. The removal of unhydrolyzed agroresidues, xylanase and xylose from the hydrolysate by two-step ultrafiltration led to the purification of XOS as confirmed by TLC as well as HPLC analysis. PMID:25553569

  3. Evaluation of the efficacy of three indigenous strains of entomopathogenic nematodes from Meghalaya, India against mustard sawfly, Athalia lugens proxima Klug (Hymenoptera: Tenthredinidae)

    OpenAIRE

    Yadav, Arun K; Lalramliana

    2012-01-01

    The objective of this study was to evaluate the efficacy of three indigenous strains of entomopathogenic nematodes (EPN) from Meghalaya, India, namely Heterorhabditis indica Poinar, Karunakar and David, Steinernema thermophilum Ganguly and Singh, and Steinernema glaseri (Steiner) against the last instar larva of mustard sawfly, Athalia lugens proxima Klug, a serious pest of mustard and radish in India. The larvae of A. lugens proxima were exposed to 10, 25, 50, 75 and 100 infective juveniles ...

  4. Chimeric Lactase Capable of Spontaneous and Strong Immobilization on Cellulose and Development of a Continuous-Flow System for Lactose Hydrolysis at High Temperatures▿

    OpenAIRE

    Velikodvorskaya, G. A.; Tikhonova, T. V.; Gurvits, I. D.; Karyagina, A. S.; Lavrova, N. V.; O. V. Sergienko; Tashlitskii, V. N.; Lunina, N. A.; Lunin, V.G.

    2010-01-01

    Recombinant plasmids containing fusion proteins composed of two different modules were constructed and expressed in Escherichia coli. The modules encoded the lactase LacA (LacZ) from the thermophilic bacterium Thermoanaerobacter ethanolicus and the cellulase CelD, a cellulose-binding module (CBM) from Anaerocellum thermophilum. The CelD CBM provides a spontaneous and strong sorption of the fusion proteins onto a cellulose carrier. The enzymatic activities of both the free LacA protein and Lac...

  5. 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. PMID:25941022

  6. Local adaptation constrains the distribution potential of heat-tolerant Symbiodinium from the Persian/Arabian Gulf.

    Science.gov (United States)

    D'Angelo, Cecilia; Hume, Benjamin C C; Burt, John; Smith, Edward G; Achterberg, Eric P; Wiedenmann, Jörg

    2015-12-01

    The symbiotic association of corals and unicellular algae of the genus Symbiodinium in the southern Persian/Arabian Gulf (PAG) display an exceptional heat tolerance, enduring summer peak temperatures of up to 36 °C. As yet, it is not clear whether this resilience is related to the presence of specific symbiont types that are exclusively found in this region. Therefore, we used molecular markers to identify the symbiotic algae of three Porites species along >1000 km of coastline in the PAG and the Gulf of Oman and found that a recently described species, Symbiodinium thermophilum, is integral to coral survival in the southern PAG, the world's hottest sea. Despite the geographic isolation of the PAG, we discovered that representatives of the S. thermophilum group can also be found in the adjacent Gulf of Oman providing a potential source of thermotolerant symbionts that might facilitate the adaptation of Indian Ocean populations to the higher water temperatures expected for the future. However, corals from the PAG associated with S. thermophilum show strong local adaptation not only to high temperatures but also to the exceptionally high salinity of their habitat. We show that their superior heat tolerance can be lost when these corals are exposed to reduced salinity levels common for oceanic environments elsewhere. Consequently, the salinity prevailing in most reefs outside the PAG might represent a distribution barrier for extreme temperature-tolerant coral/Symbiodinium associations from the PAG. PMID:25989370

  7. SYNONYMOUS CONDON USAGE BIAS AND OVEREXPRESSION OF A SYNTHETIC xynB GENE FROM Aspergillus niger NL-1 IN Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Fei Li, Shiyi Yang,

    2012-02-01

    Full Text Available To further improve the expression level of recombinant xylanase in Pichia pastoris, the xynB gene, encoding the mature peptide from Aspergillus niger NL-1, was designed and synthesized based on the synonymous condon bias of P. pastoris and optimized G+C content. 155 nucleotides were changed, and the GC content decreased from 57.7% to 43.6%. The synthetic xynB was inserted into the pPICZaA and then integrated into P. pastoris GS115. The activity of the recombinant xylanase reached 1414.7 U/mL, induced with 0.8% methanol after 14-day cultivation at a temperature of 28oC in shake flasks, which was 267% higher than that of the native gene. Furthermore, the maximum xylanase activity of 20424.2 U/mL was obtained by high-density fermentation in a 5-L fermenter, which was the highest xylanase expression in P. pastoris yet reported. The recombinant xylanase had its optimal activity at a pH of 5.0 and temperature of 50oC. The recombinant xylanase was stable over a pH range of 4.5 to 8.0. Thus, this report provides an industrial means to produce the recombinant xylanase in P. pastoris.

  8. Ethylene biosynthesis-inducing protein from cellulysin is an endoxylanase.

    Science.gov (United States)

    Fuchs, Y; Saxena, A; Gamble, H R; Anderson, J D

    1989-01-01

    The proteinaceous ethylene biosynthesis-inducing factor (EIF) that was purified from Cellulysin was also shown to contain a xylanase activity. In all nondenaturing protein separation methods employed (Sephacryl S-200 chromatography, and preparative isoelectric focusing and agarose electrophoresis), xylanase activity copurified with the ethylene biosynthesis-inducing activity. Treatment with heat (60 degrees C) or proteases in 8 molar urea inhibited both ethylene-inducing and xylanase activities. Antibodies raised against purified EIF, which contains three polypeptides of 18, 14, and 10 kilodaltons, immunoprecipitated both ethylene biosynthesis-inducing and xylanase activities. The purified EIF contained no detectable cellulase, polygalacturonase, or protease activity. Other hydrolytic activities as estimated by using p-nitrophenyl derivatives of several sugars as substrates also were not detected. Different commercially available hydrolytic enzyme preparations were tested for both ethylene biosynthesis-inducing and xylanase activities. All enzymes tested contained xylanase activity, but only a few induced ethylene biosynthesis. Western blots of proteins separated by SDS-PAGE, using antibodies prepared against the non-denatured purified EIF, revealed two major bands of about 18 and 14 kilodaltons in EIF. These antibodies seem to be specific for these proteins from Trichoderma viride, because there was little cross-reactivity with the other proteins in Cellulysin and other commercial enzyme preparations. Based on these data, we suggest that EIF contains a specific xylanase activity which is involved in inducing ethylene biosynthesis. PMID:16666504

  9. Production and characterization of ethanol- and protease-tolerant and xylooligosaccharides-producing endoxylanase from Humicola sp. Ly01.

    Science.gov (United States)

    Zhou, Junpei; Wu, Qian; Zhang, Rui; Yang, Yuying; Tang, Xianghua; Li, Junjun; Ding, Junmei; Dong, Yanyan; Huang, Zunxi

    2013-06-28

    This paper reports the production and characterization of crude xylanase from the newly isolated Humicola sp. Ly01. The highest (41.8 U/ml) production of the crude xylanase was obtained under the optimized conditions (w/v): 0.5% wheat bran, 0.2% KH2PO4, and 0.5% peptone; initial pH 7.0; incubation time 72 h; 30°C; and 150 rpm. A considerable amount of the crude xylanase was induced using hulless barley bran or soybean meal as the carbon source, but a small amount of the enzyme was produced when supplementary urea was used as the nitrogen source to wheat bran. The crude xylanase showed apparent optimal cellulase-free xylanase activity at 60°C and pH 6.0, more than 71.8% of the maximum xylanase activity in 3.0-30.0% (v/v) ethanol and more than 82.3% of the initial xylanase activity after incubation in 3.0-30.0% (v/v) ethanol at 30°C for 2 h. The crude xylanase was moderately resistant to both acid and neutral protease digestion, and released 7.9 and 10.9 μmol/ml reducing sugar from xylan in the simulated gastric and intestinal fluids, respectively. The xylooligosaccharides were the main products of the hydrolysis of xylan by the crude xylanase. These properties suggested the potential of the crude enzyme for being applied in the animal feed industry, xylooligosaccharides production, and high-alcohol conditions such as ethanol production and brewing. PMID:23676918

  10. PRODUÇÃO, CARACTERIZAÇÃO E AVALIAÇÃO DE ENZIMAS FIBROLÍTICAS NA DIGESTIBILIDADE DA FORRAGEM DE MILHO

    OpenAIRE

    Cristine dos Santos Settimi Cysneiros; Reginaldo Nassar Ferreira; Michelly Ayres Oliveira; Adriano Oliveira Favoretto; Emmanuel Arnhold; Cirano José Ulhoa

    2013-01-01

    The objectives of this research were to produce and characterize an enzyme complex (EC), using the fungus Humicola grisea, and evaluate its effect on true digestibility of forage maize dry matter. We observed that the fungus produced cellulase, b-glucosidase and xylanase enzymes. The cellulase and xylanase activities were high at the temperature of 50° C. The optimum temperature of β-glucosidase was between 50 and 60° C. The optimum pH of cellulase and xylanase enzyme was 6.0. As for β-glucos...

  11. Enzyme supplementation to improve the nutritional value of fibrous feed ingredients in swine diets fed in dry or liquid form.

    Science.gov (United States)

    Moran, K; de Lange, C F M; Ferket, P; Fellner, V; Wilcock, P; van Heugten, E

    2016-03-01

    This study evaluated the effect of xylanase supplementation (with or without), feeding method (dry or liquid), and feedstuff (corn distiller's dried grains with solubles [DDGS] or wheat middlings) on apparent total tract digestibility (ATTD) and apparent ileal digestibility (AID) of GE and nutrients, intestinal morphology, ileal and cecal pH, and VFA concentrations. Sixty-four growing pigs (25.87 ± 0.38kg initial BW) were blocked by BW and sex and randomly assigned to 8 dietary treatments. Within each feedstuff, diets were fed either liquid or dry, without or with xylanase (24,000 birch xylan units/kg feed), for 16 d. Diets contained 3.32 and 3.19 Mcal/kg ME for DDGS- and wheat middlings-based diets, respectively. Pigs were fed restricted at 3 times maintenance ME requirements. Liquid diets were prepared by steeping DDGS or wheat middlings with water (1:3, wt/vol) with or without xylanase for 24 h followed by mixing with a basal ingredient mixture and water to achieve a final ratio of 1:2.5 (wt/vol). During steeping of wheat middlings, some fiber degradation occurred. When xylanase was added in dry wheat middlings diets, AID of GE ( wheat middlings diets without xylanase (64.50 vs. 54.67% and 52.88 vs. 31.69%, respectively), but supplementation of xylanase did not impact AID of GE and NDF when liquid wheat middlings diets were fed. Xylanase in liquid DDGS diets increased ( wheat middlings diets improved ( wheat middlings diets without xylanase (80.37 vs. 78.07% and 80.23 vs. 77.94%, respectively). However, there was no effect of xylanase in DDGS diets. Pigs fed DDGS diets had greater concentrations of butyrate in the cecum ( = 0.001) than pigs fed wheat middlings diets (27.6 vs. 20.4 mmol/L). Pigs fed DDGS diets with xylanase had deeper crypts ( wheat middlings diets. Results suggest that liquid feeding and xylanase supplementation had limited potential to enhance nutrient digestibility in pigs fed DDGS-based diets. However, xylanase supplementation in dry wheat

  12. Identification of thermostable beta-xylosidase activities produced by Aspergillus brasiliensis and Aspergillus niger

    DEFF Research Database (Denmark)

    Pedersen, Mads; Lauritzen, H.K.; Frisvad, Jens Christian; Meyer, Anne S.

    2007-01-01

    Twenty Aspergillus strains were evaluated for production of extracellular cellulolytic and xylanolytic activities. Aspergillus brasiliensis, A. niger and A. japonicus produced the highest xylanase activities with the A. brasiliensis and A. niger strains producing thermostable beta-xylosidases. The...

  13. Identification of thermostable β-xylosidase activities produced by Aspergillus brasiliensis and Aspergillus niger

    DEFF Research Database (Denmark)

    Pedersen, Mads; Lauritzen, Henrik Klitgaard; Frisvad, Jens Christian; Meyer, Anne Boye Strunge

    2007-01-01

    Twenty Aspergillus strains were evaluated for production of extracellular cellulolytic and xylanolytic activities. Aspergillus brasiliensis, A. niger and A. japonicus produced the highest xylanase activities with the A. brasiliensis and A. niger strains producing thermostable beta-xylosidases. The...

  14. Construction and Identification of Plasmid pTA-TUB2

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    An about 1.40 Kb target gene fragment was yielded by PCR amplification with the plasmid pRB 129,which was identified by restriction enzyme digestion that the PCR product was TU B2 gene.The gene was digested by the restriction enzyme and was linked with pTA plasmid to construct pTA-TU B2 plasmid.The plasmid was transformed into Chaetomium spp.by PEG method and the transformation rate was 27/(2×105) and it is nine times higher than that of pRB 129.The transformants can grow on the PDA containing 1 000 μg*mL-1 carbendazim,which is 1 000 times higher than the original Chaetomium spp.The resistance was stable after 10 times transfer on non-selective medium.

  15. Seleksi dan Identifikasi Bakteri Alkalifilik Penghasil Xilanase dari Tanah Bukit Krakitan, Bayat, Klaten

    Directory of Open Access Journals (Sweden)

    SURANTO

    2007-05-01

    Full Text Available Xylanase producing alkaliphilic bacteria are bacteria that can grow well in alkaline environment. They can produce xylanase that optimum at high pH to hydrolyze xylans to be xylooligosaccharides and xylose. The calcareous soil with pH more than 7,3 can be xylanase producing alkaliphilic bacteria nature habitat. The aims of this research were (1 to isolate xylanase producing alkaliphilic bacteria isolated from calcareous soil in limestone hill area Krakitan, Bayat, Klaten and (2 to determine the kind of xylanase producing alkaliphilic bacteria which has high xylanolytic activity. For isolation and selection of xylanase producing alkaliphilic bacteria, the alkaline medium pH 9-10 were used, and 0,5 % oat spelt xylan was added. The calcareous soil was taken from limestone hill area and then was filtered and inoculated on to alkaline medium. After that, they were incubated in shaker incubator for 3 days at 150 rpm in 30-37°C. Then, the enrichment sample was grown in the same medium, that 1,8% agar was added. The colonies that grew and produced clearing zone which had the highest xylanolytic activity were chosen then characterized and identified. The growth ability isolates were examined in xylan medium at pH 7, 8, 9, 10, and 11 in order to decide bacteria of fakultative or obligate alkaliphilic. Xylanase producing alkaliphilic bacteria could be isolated from calcareous soil in limestone hill area Krakitan, Bayat, Klaten. The identified 10 xylanase producing alkaliphilic bacteria that had xylanolytic activity were Pseudomonas HT-1a, Pseudomonas HT-1b, Bacillus HT-2a, Bacillus HT-2d, Flavobacterium rigense HT-3c, Micrococcus luteus HT-3d, Paracoccus alcaliphilus HT-5c, Alcaligenes HT-4c, Alcaligenes HT-5a, and Alcaligenes HT-5b.

  16. Hemicellulases in the bleaching and characterisation of kraft pulps. Doctoral thesis

    Energy Technology Data Exchange (ETDEWEB)

    Suurnaekki, A.

    1996-03-01

    Xylanase-aided bleaching of kraft pulps is the major industrial application of hemicellulases in pulp processing. In addition to process aids, hemicellulases have recently also been shown to be promising tools in fibre analytics. In this work, the role of xylanase and mannanase pretreatments in the bleaching of softwood pulps produced by different sulphate cooking methods was studied. In addition, the action of hemicellulases in kraft fibres was characterized and exploited in the analysis of the suface composition of kraft pulps.

  17. The genes for three xylan-degrading activities from Bacteroides ovatus are clustered in a 3.8-kilobase region.

    OpenAIRE

    Whitehead, T. R.; Hespell, R B

    1990-01-01

    Genes coding for three xylan-degrading activities, xylanase, xylosidase, and arabinosidase, were simultaneously cloned from the colonic anaerobic organism Bacteriodes ovatus. The genes for the three enzymes were located on a 3.8-kilobase EcoRI genomic insert and were cloned by using plasmid pUC18. All three activities were expressed in Escherichia coli JM83, and all were cell associated. Expression of the xylanase gene was independent from expression of the xylosidase and arabinosidase genes,...

  18. Effect of Chlorine Dioxide Gas on Fungi and Mycotoxins Associated with Sick Building Syndrome

    OpenAIRE

    Wilson, S. C.; Wu, C; Andriychuk, L. A.; Martin, J. M.; Brasel, T. L.; Jumper, C. A.; Straus, D C

    2005-01-01

    The growth of indoor molds and their resulting products (e.g., spores and mycotoxins) can present health hazards for human beings. The efficacy of chlorine dioxide gas as a fumigation treatment for inactivating sick building syndrome-related fungi and their mycotoxins was evaluated. Filter papers (15 per organism) featuring growth of Stachybotrys chartarum, Chaetomium globosum, Penicillium chrysogenum, and Cladosporium cladosporioides were placed in gas chambers containing chlorine dioxide ga...

  19. Biological Control of Olive Green Mold in Agaricus bisporus Cultivation

    OpenAIRE

    Tautorus, T. E.; Townsley, P. M.

    1983-01-01

    Successful methods to control the damaging weed mold Chaetomium olivaceum (olive green mold) in mushroom beds are not presently known. An attempt was made to control C. olivaceum by biological means. A thermophilic Bacillus sp. which showed dramatic activity against C. olivaceum on Trypticase soy agar (BBL Microbiology Systems)-0.4% yeast extract agar plates was isolated from commercial mushroom compost (phase I). When inoculated into conventional and hydroponic mushroom beds, the bacillus no...

  20. Production of mycotoxins on artificially and naturally infested building materials

    DEFF Research Database (Denmark)

    Nielsen, Kristian Fog; Gravesen, S.; Nielsen, P.A.;

    1999-01-01

    In this study, the ability to produce mycotoxins during growth on artificially infested building materials was investigated for Penicillium chrysogenum, Pen. polonicum, Pen. brevicompactum, Chaetomium spp., Aspergillus ustus, Asp. niger, Ulocladium spp., Alternaria spp., and Paecilomyces spp., all......., alternariol and alternariol monomethyl ether were detected. From Ulocladium spp., Paecilomyces spp., and Asp. ustus no known mycotoxins were detected, although the latter two are known mycotoxin producers. Asp. niger produced several naphtho-gamma-pyrones and tetra-cyclic compounds. All investigated species...

  1. Osmoregulatory and tegumental ultrastructural damages to protoscoleces of hydatid cysts Echinococcus granulosus induced by fungal endophytes

    OpenAIRE

    Verma, Vijay C; Gangwar, Mayank; Nath, Gopal

    2013-01-01

    Characteristic ultrastructural changes were observed when protoscoleces of hydatid cysts Echinococcus granulosus was treated with extract of endophytic fungi Eupenicillium and Chaetomium sp. isolated from Azadirachta indica and Piper longum plants respectively. A sharp decrease in viability of protoscoleces was observed after 6 h of incubation with fungal extracts. The ultrastructural changes included rosteller disorganization, loss of hooks and shedding of the microtriches of scolex region. ...

  2. Dicty_cDB: Contig-U06884-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available CP000866_444( CP000866 |pid:none) Nitrosopumilus maritimus SCM1, c... 35 0.84 CP000477_476( CP000477 |pid:none) Methanosaeta thermo...e) Symbiobacterium thermophilum IA... 33 2.4 CP000919_324( CP000919 |pid:none) Streptococcus pneumon...e-131 3 ( EC826426 ) SME00009514 esmbsro2 Sawyeria marylandensis cDNA,... 74 4e-19 2 ( EU494202 ) Chymomyza procnemis voucher... 107543 cytochrome b (cyt... 46 1.1 1 ( EF056697 ) Philaccolilus bellissimus voucher NHM-IR65 1...... 38 0.099 CP000575_43( CP000575 |pid:none) Staphylothermus

  3. Dicty_cDB: Contig-U15121-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available U262054 ) Dictyostelium discoideum vegetative cDNA clone:VS... 1057 0.0 2 ( AU269272 ) Dictyostelium discoideum vegetative...( AU261749 ) Dictyostelium discoideum vegetative cDNA clone:VS... 870 0.0 2 ( C90128 ) Dictyostelium discoid...AU262428 ) Dictyostelium discoideum vegetative cDNA clone:VS... 547 e-156 2 ( AU071376 ) Dictyostelium disco...CU928178_107( CU928178 |pid:none) Zygosaccharomyces rouxii strain ... 53 1e-05 AC116032_6( AC116032 |pid:none) Dictyostelium disco...um thermophilum IA... 43 0.014 L36204_1( L36204 |pid:none) Dictyostelium discoideum c

  4. Ancestral genetic diversity associated with the rapid spread of stress-tolerant coral symbionts in response to Holocene climate change

    OpenAIRE

    Hume, Benjamin C C; VOOLSTRA, CHRISTIAN R.; Arif, Chatchanit; D'Angelo, Cecilia; Burt, John A.; Eyal, Gal; Loya, Yossi; Wiedenmann, Jörg

    2016-01-01

    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 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 th...

  5. Dicty_cDB: Contig-U16340-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 11289 ) Oryza glaberrima clone OG_BBa0090P20, *** SEQUENC... 36 3.2 5 ( DL174241 ) Methods for Identifying t...IYYKFVPKLKSQPKPSSPT Frame C: dn*kvisnie*klyiklslllnhlnktlnqtvklkhhikmiynhqmnndmdpielvk*li fvelelvyfykdifqlql...35 3.7 CP001393_946( CP001393 |pid:none) Anaerocellum thermophilum DSM 67... 35 4.8 (Q2IA00) RecName: Full=Sperm flagellar protein...Number of HSP's successfully gapped: 7 Length of query: 372 Length of database: 1,061,185,681 Length adjustment: 130 Effective leng... Effective search space: 153981560722 Effective search space used: 153981560722 Neighboring words threshold: 12 Window for mul

  6. Dicty_cDB: Contig-U07109-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available -17 CP000852_1186( CP000852 |pid:none) Caldivirga maquilingensis IC-16... 82 3e-1... |pid:none) Rhodococcus jostii RHA1, comple... 39 0.15 CP000852_1418( CP000852 |pid:none) Caldivirga maqui...Anaerocellum thermophilum DSM 67... 39 0.26 CP000852_1427( CP000852 |pid:none) Caldivirga maquilingensis IC-...p. TP009 clone 4 seque... 37 0.99 CP000852_1423( CP000852 |pid:none) Caldivirga maquilingensis IC-16... 37 0...2 |pid:none) Caldivirga maquilingensis IC-16... 43 0.014 AP008226_643( AP008226 |

  7. 黑曲霉固体发酵产木聚糖酶的响应面优化设计及其酶学性质的研究%Optimization of Solid State Fermentation Conditions for Xylanase Production by A spergillus niger Using Response Surface Methodology and the Enzymatic Properties

    Institute of Scientific and Technical Information of China (English)

    刘明启; 关荣发; 陈文伟

    2010-01-01

    本研究采用响应面法的中心组合对影响黑曲霉(aspergillus niger JL 15)固体发酵产木聚糖酶的条件进行了优化.以橘皮粉为基质,补充碳源、氮源,以及含水量和发酵时间对木聚糖酶产量具有显著影响(P<0.05).模拟二次多项式回归预测模型,并建立自变量与响应值的回归方程,获得各因素的最佳水平,即:甘油、硫酸铵的添加量分别为4.2%和3.1%,含水量为61%,发酵时间为73.4 h,木聚糖酶活性最大预测值达922.9 U/g干发酵产物,验证值为917.7 U/g干发酵产物,高出基础培养基酶活3.2倍.酶学性质分析研究表明,黑曲霉木聚糖酶(XylA)的最适温度和最适pH分别为55oC和pH 5.0.XylA的K_m和V_(max)值分别为9.24 mg/mL和54.05 μmol/min/mL.Mn~(2+)、Zn~(2+)和斗和Mg~(2+)对XylA活性具有促进作用,而Fe~(3+)和cu~(2+)对XylA活性具有明显抑制作用.HPLC分析结果表明,XylA的水解桦木木聚糖和麸皮不溶性木聚糖的产物均为木糖至木六糖,主要产物为木三糖.

  8. Cellulase and xylanase productions by isolated Amazon Bacillus strains using soybean industrial residue based solid-state cultivation Produção de celulase e xilanase por cepas de Bacillus isoladas da Amazônia em culivo semi-sólido utilizando resíduo da indústria da soja

    OpenAIRE

    Júlio X. Heck; Plinho F. Hertz; AYUB Marco A.Z.

    2002-01-01

    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 ...

  9. Starch digestibility, energy utilization, and growth performance of broilers fed corn-soybean basal diets supplemented with enzymes.

    Science.gov (United States)

    Stefanello, C; Vieira, S L; Santiago, G O; Kindlein, L; Sorbara, J O B; Cowieson, A J

    2015-10-01

    A study was conducted to evaluate the effects of dietary α-amylase and β-xylanase supplementation of corn-soy diets, formulated with or without supplemental phytase, on growth performance, energy utilization, and starch digestibility in broiler chickens. A total of 336 slow-feathering, Cobb × Cobb 500 male broilers were randomly distributed to 6 treatments having 8 replicates of 7 birds each. Birds were fed a common starter diet to d 14 post-hatch (3,050 kcal/kg AMEn, 21.7% CP, 1.05% Ca, and 0.53% nPP). The experimental diets were provided afterwards until d 25. A 2 × 3 factorial arrangement of 2 control diets (basal = corn-soy diet without added phytase or PHY = corn-soy diet formulated with 1,000 phytase units/kg) and 3 carbohydrase supplementations (0, 80 kilo-Novo α-amylase units/kg, or 80 kilo-Novo α-amylase units/kg + 100 fungal β-xylanase units/kg) was used from d 14 to 25. Excreta were collected from 21 to 24 d and all birds were euthanized at 25 d for jejunum and ileum content collection. Samples of feed, excreta, and jejunal and ileal digesta were analyzed for determination of total tract retention and ileal apparent digestibility. No interactions between diet and carbohydrase were observed. Broilers fed diets formulated with phytase or supplemented with amylase + xylanase had higher BW gain (BWG) and lower FCR (P amylase and amylase + xylanase, respectively. Starch digestibility in the jejunum and ileum was increased (P amylase + xylanase. Results from this experiment show that corn-soy diets having phytase and supplemented with amylase and xylanase led to increased growth performance, AMEn, and starch digestibility in broilers. Furthermore, the efficacy of exogenous amylase and xylanase was independent of the presence of microbial phytase. PMID:26316335

  10. Production of xylanolytic enzymes by Aspergillus terricola in stirred tank and airlift tower loop bioreactors.

    Science.gov (United States)

    Michelin, Michele; Polizeli, Maria de Lourdes Teixeira de Moraes; Silva, Daniel Pereira da; Ruzene, Denise Santos; Vicente, António Augusto; Jorge, João Atílio; Terenzi, Héctor Francisco; Teixeira, José António

    2011-12-01

    Fungi producing high xylanase levels have attracted considerable attention because of their potential industrial applications. Batch cultivations of Aspergillus terricola fungus were evaluated in stirred tank and airlift bioreactors, by using wheat bran particles suspended in the cultivation medium as substrate for xylanase and β-xylosidase production. In the stirred tank bioreactor, in physical conditions of 30°C, 300 rpm, and aeration of 1 vvm (1 l min⁻¹), with direct inoculation of fungal spores, 7,475 U l⁻¹ xylanase was obtained after 36 h of operation, remaining constant after 24 h. In the absence of air injection in the stirred tank reactor, limited xylanase production was observed (final concentration 740 U l⁻¹). When the fermentation process was realized in the airlift bioreactor, xylanase production was higher than that observed in the stirred tank bioreactor, being 9,265 U l⁻¹ at 0.07 vvm (0.4 l min⁻¹) and 12,845 U l⁻¹ at 0.17 vvm (1 l min⁻¹) aeration rate. PMID:21626207

  11. ISOLATION AND IDENTIFICATION OF XYLANOLYTIC ENZYME FROM AN EFFECTIVE STRAIN BACILLUS LICHENIFORMIS ISOLATED FROM THE DECAYING WOOD

    Directory of Open Access Journals (Sweden)

    Sarika Chaturvedi

    2013-12-01

    Full Text Available A new species of Bacillus licheniformis produced extracellular xylanase under submerged fermentation when wheat bran is used as carbon source. The xylan is the most common hemicellulosic polysaccharide in food industry and agricultural wastes, comprising a backbone of xylose residues linked by β-1,4 glycosidic bonds. Bacillus licheniformis has been shown to be a promising organism for enhanced production of xylanases & β-xylosidase under submerged fermentation (SmF. The optimization of cultural conditions and carbon, nitrogen sources for enzymes production. The bacterial strain Bacillus licheniformis was cultivated using as substrate xylan, wheat bran, corn straw, corncob, and sugarcane bagasse. Wheat bran has been a good xylanase (16.8U/ml & β xylosidase (5.6U/ml activity after 48h of fermentation. Maximum enzyme activity was observed in xylan as carbon source and peptone as nitrogen source. Both crude enzymes were characterized and a bacterial xylanase shows optimum pH for xylanase activity at 6.5 & β xylosidase were found to be 6.0. The optimum temperatures were 450C for both and they were thermally stable up to 500C. The parameters of Vmax and Km obtained using Line weaver-Burk plot method were 277.7μmol / min/mg and 5.26 mg /L correspondingly

  12. Pantoea stewartii subsp. stewartii produces an endoglucanase that is required for full virulence in sweet corn.

    Science.gov (United States)

    Mohammadi, Mojtaba; Burbank, Lindsey; Roper, M Caroline

    2012-04-01

    Pantoea stewartii subsp. stewartii, a xylem-dwelling bacterium, is the causal agent of Stewart's wilt and blight of sweet corn. The goal of this study was to characterize the only gene in the P. stewartii subsp. stewartii genome predicted to encode an endoglucanase (EGase); this gene was designated engY. Culture supernatants from P. stewartii subsp. stewartii and Escherichia coli expressing recombinant EngY protein possessed both EGase and xylanase activities. Deletion of engY abolished EGase and xylanase activity, demonstrating that EngY appears to be the major EGase or xylanase produced by P. stewartii subsp. stewartii. Most importantly, our results show that EngY contributes to movement in the xylem and disease severity during the wilting phase of Stewart's wilt but is not required for water-soaked lesion formation. PMID:22122328

  13. Exploring the Plant–Microbe Interface by Profiling the Surface-Associated Proteins of Barley Grains

    DEFF Research Database (Denmark)

    Sultan, Abida; Andersen, Birgit; Svensson, Birte;

    2016-01-01

    Cereal grains are colonized by a microbial community that actively interacts with the plant via secretion of various enzymes, hormones, and metabolites. Microorganisms decompose plant tissues by a collection of depolymerizing enzymes, including β-1,4-xylanases, that are in turn inhibited by plant......-associated proteins and xylanolytic activities of two barley cultivars. The surface-associated proteome was dominated by plant proteins with roles in defense and stress-responses, while the relatively less abundant microbial (bacterial and fungal) proteins were involved in cell-wall and polysaccharide degradation and...... included xylanases. The surface-associated proteomes showed elevated xylanolytic activity and contained several xylanases. Integration of proteomics with enzyme assays is a powerful tool for analysis and characterization of the interaction between microbial consortia and plants in their natural environment....

  14. 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

    DEFF Research Database (Denmark)

    Schiøtt, Morten; De Fine Licht, Henrik H; Lange, Lene; Boomsma, Jacobus J

    2008-01-01

    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...

  15. Controlled enzyme catalyzed heteropolysaccharide degradation

    DEFF Research Database (Denmark)

    Rasmussen, Louise Enggaard

    The work presented in this PhD thesis has provided a better understanding of the enzyme kinetics and quantitative phenomena of the hydrolysis of xylan substrates by selected pure enzyme preparations. Furthermore, the options for producing specific substituted xylooligosaccharides from selected...... substrates by specific xylanase treatment have been examined. The kinetics of the enzymatic degradation of water-extractable wheat arabinoxylan (WE-AX) during designed treatments with selected monocomponent enzymes was investigated by monitoring the release of xylose and arabinose. The results of different...... effects between -xylosidase and the α-L-arabinofuranosidases on the xylose release were low as compared to the effect of xylanase addition with β-xylosidase, which increased the xylose release by ~25 times in 30 minutes. At equimolar addition levels of the four enzymes, the xylanase activity was thus rate...

  16. 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

  17. Inclusion of different exogenous fibrolytic enzymes to dry jowar fodder and their effect on in vitro total gas production

    Directory of Open Access Journals (Sweden)

    S.H. Sipai

    2013-09-01

    Full Text Available Aim: Our objective was to estimate in-vitro gas production from dry jowar fodder added with differentconcentrations of exogenous fibrolytic enzymes (EFEs like neutral cellulase and fungal xylanase.Materials and Methods: 34 different samples of dry jowar fodder were prepared according to differentconcentrations of neutral cellulase, fungal xylanase and neutral cellulase + fungal xylanase (1:1. Sample notcontaining any enzymes was considered as the control group. These 34 samples were subjected to further in vitrogas production analysis.Results: Statistically, significantly higher (P<0.05 potential gas production was recorded for 0.7 % at 6 hr period,0.7 % at 12 hr period, 0.7 %, 0.8 % at 18 hr period and 0.7 %, 0.8 % at 24 hr period in the samples treated withneutral cellulase. Significantly higher potential gas production was recorded for 0.5 %, 0.8 % at 6 hr period, 0.5 %,0.6 %, 0.8 % at 12 hr period, 0.8 % at 18 hr period and 0.5 %, 0.6 %, 0.8 % at 24 hr period in the samples treated withfungal xylanase. Significantly higher potential gas production was recorded for 0.6 %, 0.6 %, 0.8 % at 6 hr period,0.6 %, 0.8 % at 12 hr period, 0.6 %, 0.8 % at 18 hr period and 0.6 %, 0.8 % at 24 hr period in the samples treated withmixture of neutral cellulase + fungal xylanase (1:1.Conclusion: Addition of neutral cellulase and fungal xylanase into the samples of dry jowar fodder increased invitro total potential gas production. EFEs increase substrate degradation and there by improve the nutritive value ofdry jowar fodder.

  18. Mixed Enzyme Systems for Delignification of Lignocellulosic Biomass

    Directory of Open Access Journals (Sweden)

    Elisa M. Woolridge

    2014-01-01

    Full Text Available The application of enzymes such as laccase and xylanase for the preparation of cellulose from lignocellulosic material is an option for those industries seeking to reduce the use of chlorine-containing bleach agents, thus minimizing the environmental impact of their processes. Mixed hydrolytic and oxidative enzyme systems have been well described in the context of biopulping, and thus provide good precedent regarding effectiveness, despite the susceptibility of xylanase to inactivation by laccase-generated oxidants. This paper examines the progress towards development of sequential and simultaneous mixed enzyme systems to accomplish delignification.

  19. Purification and characterization of two 1,4-beta-xylan endohydrolases from the rumen fungus Neocallimastix frontalis.

    OpenAIRE

    Gomez de Segura, B; Fevre, M.

    1993-01-01

    Two beta-endoxylanases produced by Neocallimastix frontalis have been purified by ammonium sulfate precipitation, gel filtration, and ion-exchange chromatography. Xylanase I is a nonglycosylated protein with an apparent molecular mass of 45 kDa. Xylanase II is a glycoprotein with an apparent molecular mass of 70 kDa. The pH optima of these enzymes were 5.5 and 6, respectively, and the temperature optimum was 55 degrees C for each enzyme. The endo mode of action of the enzymes was revealed by ...

  20. Analysis of an Effective Antibiotic (Chaetomacin) Isolated from a Thermophilic Bacillus sp. Against Olive Green Mold

    OpenAIRE

    Tautorus, T. E.; Townsley, P. M.

    1984-01-01

    Successful methods to control the damaging weed mold Chaetomium olivaceum (olive green mold) in mushroom beds are not known. An effective antibiotic (named chaetomacin) against C. olivaceum was isolated from a thermophilic Bacillus sp. This compound was shown to be an extremely potent and stable antibiotic, effective over a wide range of both pH (2 to 10) and temperature (−15 to 150°C). Chaetomacin is soluble in most polar solvents and insoluble in nonpolar solvents. It is produced only at me...

  1. Phyllosphere mycobiota on garden ponds plants

    Directory of Open Access Journals (Sweden)

    Maria Kowalik

    2013-12-01

    Full Text Available Investigations were conducted on calamus, common cattail, soft rush, yellow iris and white water lily plants in twenty ponds in Malopolska and Podkarpacie Regions. Mycobiota existing in the phyllosphere caused discolouring and necroses of leaves and shoots. 88 species of mycobiota were identified and isolated from the diseased tissues. Dominant were Alternaria alternata, Epicoccum nigrum and Isaria farinosa. Fungi of genera: Aspergillus, Botrytis, Chaetomium, Cladosporium, Fusarium, Ilyonectria, Mortierella, Mucor, Penicillium, Phialophora, Phoma, Pleustomophora, Sordaria, Trichoderma and Umbelopsis were also numerous. The monophagous and the polyphagous were identified.

  2. Effect of vapor phase corrosion inhibitor on microbial corrosion of aluminum alloys.

    Science.gov (United States)

    Yang, S S; Ku, C H; Bor, H J; Lin, Y T

    1996-02-01

    Vapor phase corrosion inhibitors were used to investigate the antimicrobial activities and anticorrosion of aluminum alloy. Aspergillus flavus, A. niger, A. versicolor, Chaetomium globosum and Penicillium funiculosum had moderate to abundant growth on the aluminum alloy AA 1100 at Aw 0.901, while there was less growth at Aw 0.842. High humidity stimulated microbial growth and induced microbial corrosion. Dicyclohexylammonium carbonate had a high inhibitory effect on the growth of test fungi and the microbial corrosion of aluminum alloy, dicyclohexylammonium caprate and dicyclohexylammonium stearate were the next. Aluminum alloy coating with vapor phase corrosion inhibitor could prevent microbial growth and retard microbial corrosion. PMID:10592784

  3. Biodiversity of endophytic fungi from seven herbaceous medicinal plants of Malnad region, Western Ghats, southern India

    Institute of Scientific and Technical Information of China (English)

    B. Shankar Naik; M. Krishnappa; Y. L. Krishnamurthy

    2014-01-01

    A total of 3611 fungal isolates were recovered from 4200 leaf segments incubated from 7 medicinal herbs during monsoon, winter and summer seasons. These fungal isolates belonged to teleomorphic Asco-mycota (23.5%), anamorphic Ascomycota producing conidiomata (17.4%), anamorphic Ascomycota without conidiomata (46.9%), Zygo-mycota (1.42%) and sterile forms (10.6%). Chaetomium globosum, As-pergillus niger, Aureobasidium pullulans, Curvularia lunata, Fusarium spp., Penicillium spp., Pestalotiopsis spp., Trichoderma viridae, Cladosporium cladosporioides, were frequently isolated from more than one host plant. The number of endophytic isolates was higher in winter than in monsoon and summer seasons.

  4. Use of agricultural by-products for the production of xylitol. I. The production of xylose

    Energy Technology Data Exchange (ETDEWEB)

    De Menezes, H.C.

    1976-01-01

    A Rhizopus species capable of converting xylan into xylose was isolated from the soil, and purified. The xylanase produced by this fungus was capable of producing xylose from corn cob, wheat bran, and rice hulls without prior extraction of the xylan.

  5. Trichoderma reesei CE16 acetyl esterase and its role in enzymatic degradation of acetylated hemicellulose

    DEFF Research Database (Denmark)

    Biely, Peter; Cziszarava, Maria; Agger, Jane W.;

    2014-01-01

    Results The combined action of GH10 xylanase and acetylxylan esterases (AcXEs) leads to formation of neutral and acidic xylooligosaccharides with a few resistant acetyl groups mainly at their non-reducing ends. We show here that these acetyl groups serve as targets for TrCE16 AcE. The most promin...

  6. 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...

  7. PRODUÇÃO, CARACTERIZAÇÃO E AVALIAÇÃO DE ENZIMAS FIBROLÍTICAS NA DIGESTIBILIDADE DA FORRAGEM DE MILHO

    Directory of Open Access Journals (Sweden)

    Cristine dos Santos Settimi Cysneiros

    2013-12-01

    Full Text Available The objectives of this research were to produce and characterize an enzyme complex (EC, using the fungus Humicola grisea, and evaluate its effect on true digestibility of forage maize dry matter. We observed that the fungus produced cellulase, b-glucosidase and xylanase enzymes. The cellulase and xylanase activities were high at the temperature of 50° C. The optimum temperature of β-glucosidase was between 50 and 60° C. The optimum pH of cellulase and xylanase enzyme was 6.0. As for β-glucosidase, the enzyme showed higher activity at pH 6.5. Cellulase remained stable for 60 minutes at 39° C. Xylanase and β-glucosidase maintained 99.2 and 88.2% of their activity at 50° C for 240 minutes, respectively. The treatments were as follows: control (10 mL of sterile water, level 1 (2.5 mL EC, level 2 (5.0 mL EC and level 3 (10 mL EC. In the digestibility experiment, there was interaction between enzyme levels and time of incubation in the rumen. The addition of 10 mL of the fibrolytic enzymes improved the digestibility at 10.58; 12.52; 9.05 and 6.81% compared to control for 12; 24; 48 and 96 hours of incubation, respectively. The fungus Humicola grisea is an enzyme producer that is important in ruminant feed.

  8. Interaction of water unextractable solids with gluten protein: Effect on dough properties and gluten quality

    NARCIS (Netherlands)

    Wang, M.; Oudgenoeg, G.; Vliet, T. van; Hamer, R.J.

    2003-01-01

    In a previous study, we have shown that water unextractable solids (WUS) interfere with gluten formation and affect the quality of the resulting gluten. In this study we aim to explain how WUS can affect the process of gluten formation. To this end, WUS were modified with NaOH, xylanase, horseradish

  9. Extracellular Hemicellulolytic Enzymes from the Maize Endophyte Acremonium zeae

    Science.gov (United States)

    The maize endophyte Acremonium zeae was examined for production of extracellular enzymes that hydrolyze cellulose and hemicellulose. The most prominent enzyme activity in cell-free culture media from A. zeae NRRL 6415 was xylanase, with a specific activity of 60 U/mg from cultures grown on crude co...

  10. Cloning and characterization of alpha-glucuronidase enzyme

    Science.gov (United States)

    Hemicellulose is the second largest source of biomass on Earth. Xylan, a polymer of beta-1,4-linked xylose residues, is a common component of hemicellulose. The enzymes xylanase and beta-xylosidase hydrolyze the xylan into xylose which can then be fermented into value-added products. However, the...

  11. Delignification outperforms alkaline extraction for xylan fingerprinting of oil palm empty fruit bunch.

    Science.gov (United States)

    Murciano Martínez, Patricia; Kabel, Mirjam A; Gruppen, Harry

    2016-11-20

    Enzyme hydrolysed (hemi-)celluloses from oil palm empty fruit bunches (EFBs) are a source for production of bio-fuels or chemicals. In this study, after either peracetic acid delignification or alkaline extraction, EFB hemicellulose structures were described, aided by xylanase hydrolysis. Delignification of EFB facilitated the hydrolysis of EFB-xylan by a pure endo-β-1,4-xylanase. Up to 91% (w/w) of the non-extracted xylan in the delignified EFB was hydrolysed compared to less than 4% (w/w) of that in untreated EFB. Alkaline extraction of EFB, without prior delignification, yielded only 50% of the xylan. The xylan obtained was hydrolysed only for 40% by the endo-xylanase used. Hence, delignification alone outperformed alkaline extraction as pretreatment for enzymatic fingerprinting of EFB xylans. From the analysis of the oligosaccharide-fingerprint of the delignified endo-xylanase hydrolysed EFB xylan, the structure was proposed as acetylated 4-O-methylglucuronoarabinoxylan. PMID:27561506

  12. Biosynthesis of xylobiose: a strategic way to enrich the value of oil palm empty fruit bunch fiber.

    Science.gov (United States)

    Lakshmi, G Suvarna; Rajeswari, B Uma; Prakasham, R S

    2012-08-01

    Xylooligosaccharides are functional foods mainly produced during the hydrolysis of xylan by physical, chemical, or enzymatic methods. In this study, production of xylobiose was investigated using oil palm empty fruit bunch fiber (OPEFB) as a source material, by chemical and enzymatic methods. Xylanase-specific xylan hydrolysis followed by xylobiose production was observed. Among different xylanases, xylanase from FXY-1 released maximum xylobiose from pretreated OPEFB fiber, and this fungal strain was identified as Aspergillus terreus and subsequently deposited under the accession Number MTCC- 8661. The imperative role of lignin on xylooligosaccharides enzymatic synthesis was exemplified with the notice of xylobiose production only with delignified material. A maximum 262 mg of xylobiose was produced from 1.0 g of pretreated OPEFB fiber using FXY-1 xylanase (6,200 U/ml) at pH 6.0 and 45° C. At optimized environment, the yield of xylobiose was improved to 78.67 g/100 g (based on xylan in the pretreated OPEFB fiber). PMID:22713984

  13. 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 xylos

  14. Trichoderma reesei XYN VI--a novel appendage-dependent eukaryotic glucuronoxylan hydrolase.

    Science.gov (United States)

    Biely, Peter; Puchart, Vladimír; Stringer, Mary Ann; Mørkeberg Krogh, Kristian B R

    2014-09-01

    Expression of a Trichoderma reesei gene coding for a putative GH30 xylanase in Aspergillus oryzae led to isolation and purification of a novel xylanase exhibiting catalytic properties different from those of the previously characterized GH30 xylanase XYN IV of T. reesei. The novel enzyme, named XYN VI, exhibited catalytic properties similar to appendage-dependent GH30 glucuronoxylanases previously recognized only in bacteria. XYN VI showed high specific activity only on xylans or xylooligosaccharides containing 4-O-methyl-D-glucuronic acid or D-glucuronic acid side substituents. The cleavage of the main chain takes place primarily at the second glycosidic linkage from the branch towards the reducing end of the polysaccharides or aldouronic acids. These catalytic properties resemble bacterial GH30 glucuronoxylanases, although the recognition of the uronic acid side chains by XYN VI is apparently based on interaction of the substrate with other amino acids. Moreover, in contrast to bacterial enzymes, XYN VI is also capable of slower but significant cleavage of unsubstituted parts of xylan and acidic xylooligosaccharides. The data point to a great catalytic diversity of xylanases produced by the most extensively studied cellulolytic fungus. PMID:25041335

  15. Delignification outperforms alkaline extraction for xylan fingerprinting of oil palm empty fruit bunch

    NARCIS (Netherlands)

    Murciano Martínez, Patricia; Kabel, Mirjam A.; Gruppen, Harry

    2016-01-01

    Enzyme hydrolysed (hemi-)celluloses from oil palm empty fruit bunches (EFBs) are a source for production of bio-fuels or chemicals. In this study, after either peracetic acid delignification or alkaline extraction, EFB hemicellulose structures were described, aided by xylanase hydrolysis. Deligni

  16. Feruloyl esterase production by Aspergillus terreus CECT 2808 and subsequent application to enzymatic hydrolysis.

    Science.gov (United States)

    Pérez-Rodríguez, N; Moreira, C D; Torrado Agrasar, A; Domínguez, J M

    2016-09-01

    Ferulic acid esterases (FAE) were produced by Aspergillus terreus CECT 2808 from vine trimming shoots (VTS) and corn cob. Later, the fungal extracts thus obtained were used to enzymatically release ferulic acid (FA) from both substrates. Our findings showed a higher FAE activity in the enzymatic extracts produced on corn cob (0.070±0.004U/mL). Nevertheless, the enzymatic extracts produced on VTS demonstrated a better performance for FA release from both corn cob (2.05±0.01mg/g) and VTS (0.19±0.003mg/g). This result was probably because of the higher xylanase/FAE ratio determined in VTS extract. Therefore, an additional assay was carried out by supplementing corn cob extract with a commercial xylanase to test the influence of FAE/xylanase ratio in FA release. The results revealed the relevance of the FAE/xylanase ratio for an optimal FA release. PMID:27444329

  17. Cost-effective production of biotechnologically important hydrolytic enzymes by Sporotrichum thermophile.

    Science.gov (United States)

    Bala, Anju; Singh, Bijender

    2016-01-01

    Economical production of xylanase and three cellulases, endo-β-1,4-glucanase (CMCase), exo-β-1,4-glucanase (FPase), β-glucosidase (BGL) was studied in submerged fermentation using cane molasses medium. A statistical optimization approach involving Plackett-Burman design and response surface methodology (RSM) resulted in the production of 72,410, 36,420, 32,420 and 5180 U/l of xylanase, CMCase, FPase and β-glucosidase, respectively. Optimization resulted in more than fourfold improvements in production of xylanolytic and cellulolytic enzymes. Scale up of enzymes production in shake flasks of varied volumes was sustainable, suggesting a good scope for large scale enzyme production. Addition of microparticles engineered fungal morphology and enhanced enzymes production. Xylanase of S. thermophile is a neutral xylanase displaying its optimal activity at 60 °C while all the cellulases are optimally active at pH 5.0 and 60 °C. The efficacy of enzyme cocktail in waste tea cup paper and rice straw hydrolysis showed that maximum sugar yield of 578.12 and 421.79 mg/g substrate for waste tea cup and rice straw, respectively, were achieved after 24 h. Therefore, concomitant production of cellulolytic and xylanolytic enzymes will be beneficial for the saccharification of lignocellulosics in generating both monomeric and oligomeric sugars for biofuels and other biotechnological applications. PMID:26581490

  18. Dicty_cDB: Contig-U11841-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available meleagridis partial mRN... 100 2e-19 U66424_1( U66424 |pid:none) Arabidopsis thalia...2 7e-11 CR548612_182( CR548612 |pid:none) Paramecium tetraurelia macronucl... 58 7e-11 AE014188_70( AE014188 |pid:none) Plas...146_1446( CP001146 |pid:none) Dictyoglomus thermophilum H-6-1... 60 3e-07 CR548612_204( CR548612 |pid:none) Paramecium tetraurelia...er chromos... 59 6e-07 E71606( E71606 )hypothetical protein PFB0765w - malaria parasite (... 59 6e-07 CP0000...: Full=Myosin heavy chai... 59 7e-07 DQ227281_1( DQ227281 |pid:none) Canis familiaris fast myosin heavy... 5

  19. AcEST: DK957554 [AcEST

    Lifescience Database Archive (English)

    Full Text Available plantlets Developmental stage gametophytes with sporophytes C...us-veneris mRNA. clone: TST39A01NGRL0028_K21. 5' end sequence. Accession DK957554 Tissue type prothallia with...|Q67SM3|Q67SM3_SYMTH Putative uncharacterized protein OS=Symbiobacterium thermophilum Align length 31 Score (bit) 35.4 E-val...ochrome c-type protein torY OS=Haemophil... 30 8.3 >sp|O73691|EGR1_CHICK Early growth response prote...SPAHTTFPSPSIATTYS 148 >sp|P44799|TORY_HAEIN Cytochrome c-type protein torY OS=Haemophilus influenzae GN=torY PE=3 SV=1 Length

  20. Dicty_cDB: Contig-U13605-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ... 82 2e-14 CP000852_493( CP000852 |pid:none) Caldivirga maquilingensis IC-167... 80 8e-14 AM8494...one) Herpetosiphon aurantiacus ATCC ... 42 0.019 CP001348_774( CP001348 |pid:none) Clostridium cellulolyticu...159437_1( AK159437 |pid:none) Mus musculus osteoclast-like cell ... 191 2e-47 M76762_1( M76762 |pid:none) Mu...J783889_1( AJ783889 |pid:none) Dascillus cervinus mRNA for S18e r... 181 2e-44 CP000499_614( CP000499 |pid:n... translocase... 41 0.042 CP001393_1648( CP001393 |pid:none) Anaerocellum thermophilum DSM 6... 4