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Sample records for chaetomium thermophilum xylanase

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

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    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.

  2. 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

    been sequenced, its carbohydrate esterases are not annotated yet. We applied peptide pattern recognition (PPR) tool for sequence analysis of the C. thermophilum genome, and 11 carbohydrate esterase genes were discovered. Furthermore, we cloned and heterologously expressed two putative acetyl xylan...

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

    DEFF Research Database (Denmark)

    Jepsen, Helene Friborg; Jensen, B.

    2004-01-01

    against high concentrations of sodium chloride nor directly linked to thermophily. In C. thermophilum var. coprophilum three different trehalose hydrolyzing activities were eluted from a mono Q anion exchange column by sodium chloride concentrations of 0.10, 0.15 and 0.24 M, respectively....

  4. Breeding of xylanase producing strain with chaetomium globosum by ultraviolet mutation%紫外诱变球毛壳霉选育木聚糖酶高产菌株

    Institute of Scientific and Technical Information of China (English)

    杨健; 姚色笛; 王颖; 王月; 张丽媛

    2011-01-01

    Xylanase has important potential applications value. In order to gain the strain which has stable genetic traits to produce xylanase, we took the xylanase activity as evaluation index, used ultraviolet mutation method, determined the time of ultraviolet irradiation by single factor experiment. By preliminary screening of Ultraviolet mutation strains in the medium containing xylan, the xylanase activity was measured by DNS methods. Screen was then repeated in the mutation strains. The highest xylanase activity mutant strains were selected to determine their activity and the genetic stability. The results of this experiment showed that the power of ultraviolet lamp was 20W, exposure distance was 30cm and exposure time was 120s. The fatality rate of chaetomium globosum was 92. 35%. Under these conditions, 15 mutant strains with higher yield of xylanase was selected. The strain Z30 - 56 had the highest xylanase activity and highest yield was 9850IU/mL, increasing 53.7% compared with the original strain. The genetics of the strain was also very stable.%木聚糖酶具有重要的潜在应用价值,为获得一株遗传性状稳定的木聚糖酶高产菌株,以木聚糖酶的酶活为评价指标,采用紫外线诱变的方法,利用单因素试验确定紫外线照射时间,通过含木聚糖的初筛培养基对紫外诱变后的菌株进行初筛,然后采用DNS法测定木聚糖酶活性对诱变菌株进行复筛.将筛选出的木聚糖酶活力最高的诱变菌株进行传代培养,测定其酶活,探讨其遗传稳定性.试验结果表明:紫外灯功率20W,照射距离30cm,照射时间120s,球毛壳霉致死率为92.35%.在此条件下进行菌株的紫外诱变获得高产木聚糖酶的正突变株15株,其中1株高产菌株Z30 - 56的酶活为9850IU/mL,比出发菌株提高53.7%,并且遗传性能稳定.

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

    DEFF Research Database (Denmark)

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

    2010-01-01

    The genes encoding the two endo-xylanases XynA and XynB from the thermophilic bacterium Dictyoglomus thermophilum were codon optimized for expression in plants. Both xylanases were designed to be constitutively expressed under the control of the CaMV 35S promoter and targeted to the apoplast. Tra...

  6. Chromones from an ascomycete,Chaetomium aureus

    Institute of Scientific and Technical Information of China (English)

    Li Mei Li; Qiang Zou; Guo You Li

    2010-01-01

    A novel chromone,named chaetoaurin (1),along with six known chromone derivatives (2-7),was isolated from the ethyl acetate extract of a solid-state fermented culture of Chaetomium aureus.Their structures were elucidated by extensive spectral analysis.All of these compounds were reported from C.aureus for the first time.

  7. Fungi of Delhi XXXIII. Chaetomium putrefactus sp.n.

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    Rani Grupta

    2014-08-01

    Full Text Available Chaetomium putrefactus has been described as a new species. It has smaller and sparser hairs and ellipto-fusoid ascospores. We have isolated and described several species of Chaetomium from living and dead leaves of various plants. One species isolated from decaying leaves of Corchorus olitorius appeared to be interesting and new. It is characterized by smaller and fewer hairs and ellipso-fusoid ascospores.

  8. [Occupational allergies to xylanases].

    Science.gov (United States)

    van Kampen, V; Merget, R; Brüning, T

    2004-02-01

    The exposure against enzyme dusts have long been known to cause occupational allergies. In the 1960s an increasing number of occupational allergies in the detergent industry were observed. In this context the high sensitization potential of enzyme dusts attracted attention. The present evaluation of literature data confirms that this is also true for xylanases. These frequently used industrial enzymes belong to the hemicellulases and are mostly of fungal origin. Several cases of specific airway sensitization caused by xylanases or other hemicellulases are verified by a number of case reports and cross sectional studies. As symptoms, results of skin prick tests, detection of specific IgE-antibodies and results of specific bronchoprovocation tests are consistent, an immunologic mechanism can be assumed.

  9. Analysis of the tryptophanase expression in Symbiobacterium thermophilum in a coculture with Geobacillus stearothermophilus.

    Science.gov (United States)

    Watsuji, Tomo-O; Takano, Hideaki; Yamabe, Tomoya; Tamazawa, Satoshi; Ikemura, Hiroka; Ohishi, Takanori; Matsuda, Tohyo; Shiratori-Takano, Hatsumi; Beppu, Teruhiko; Ueda, Kenji

    2014-12-01

    The tryptophanase-positive Symbiobacterium thermophilum is a free-living syntrophic bacterium that grows effectively in a coculture with Geobacillus stearothermophilus. Our studies have shown that S. thermophilum growth depends on the high CO2 and low O2 condition established by the precedent growth of G. stearothermophilus. The use of an anoxic atmosphere containing high CO2 allows S. thermophilum to grow independently of G. stearothermophilus, but the cellular yield is ten times lower than that achieved in the coculture. In this study, we characterized the coculture-dependent expression and activity of tryptophanase in S. thermophilum. S. thermophilum cells accumulated a marked amount of indole in a coculture with G. stearothermophilus, but not in the bacterium's pure culture irrespective of the addition of tryptophan. S. thermophilum cells accumulated indole in its pure culture consisting of conditioned medium (medium supplied with culture supernatant of G. stearothermophilus). Proteomic analysis identified the protein specifically produced in the S. thermophilum cells grown in conditioned medium, which was a tryptophanase encoded by tna2 (STH439). An attempt to isolate the tryptophanase-inducing component from the culture supernatant of G. stearothermophilus was unsuccessful, but we did discover that the indole accumulation occurs when 10 mM bicarbonate is added to the medium. RT-PCR analysis showed that the addition of bicarbonate stimulated transcription of tna2. The transcriptional start site, identified within the tna2 promoter, was preceded by the -24 and -12 consensus sequences specified by an alternative sigma factor, σ(54). The evidence suggests that the transcription of some genes involved in amino acid metabolism is σ(54)-dependent, and that a bacterial enhancer-binding protein containing a PAS domain controls the transcription under the presence of high levels of bicarbonate.

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

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    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.

  11. Chaetomium and Stachybotrys in water-damaged buildings

    DEFF Research Database (Denmark)

    Andersen, Birgitte; Lewinska, Anna Malgorzata; Nielsen, Jakob Blæsbjerg

    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....

  12. Bacillus thermophilum sp. nov., isolated from a microbial fuel cell.

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    Tang, Jia; Yang, Guiqin; Wen, Junlin; Yu, Zhen; Zhou, Shungui; Liu, Zhi

    2014-09-01

    A novel thermophilic, Gram-staining positive bacterium, designated DX-2(T), was isolated from the anode biofilm of a microbial fuel cell. Cells of the strain were oxidase positive, catalase positive, facultative anaerobic, motile rods. The isolate grew at 30-60 °C (optimum 50 °C) and pH 5-9 (optimum pH 8-8.5). The pairwise 16S rRNA gene sequence similarities showed that strain DX-2(T) was most closely related to Bacillus fumarioli LMG 17489(T) (96.2 %), B. firmus JCM 2512(T) (96.0 %) and B. foraminis DSM 19613(T) (95.7 %). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain DX-2(T) formed a cluster with B. smithii (95.5 %) and B. infernus (94.9 %). The genomic G+C content of DX-2(T) was 43.7 mol%. The predominant respiratory quinone was MK-7. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and unknown phospholipids. The major cellular fatty acid was iso-C16:0. Based on its phenotypic characteristics, chemotaxonomic features, and results of phylogenetic analysis, the strain was identified to represent a distinct novel species in the genus Bacillus, and the name proposed is B. thermophilum sp. nov. The type strain is DX-2(T) (=CCTCC AB2012194(T) = KCTC 33128(T)).

  13. 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...... was found, but no single stranded intermediates, characteristic of rolling circle replication, were found on Southern blots. The larger plasmid, pBAL, was found to be a 8294 bp plasmid with a GC content of 39%. It revealed 17 ORFs, of which three showed similarity at the amino acid (aa) level to known...

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

    NARCIS (Netherlands)

    Wiegant, W.M.

    1992-01-01

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

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

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    Fliss Ismaïl

    2007-08-01

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

  16. Efficacy of Chaetomium Species as Biological Control Agents against Phytophthora nicotianae Root Rot in Citrus.

    Science.gov (United States)

    Hung, Phung Manh; Wattanachai, Pongnak; Kasem, Soytong; Poeaim, Supattra

    2015-09-01

    Thailand is one of the largest citrus producers in Southeast Asia. Pathogenic infection by Phytophthora, however, has become one of major impediments to production. This study identified a pathogenic oomycete isolated from rotted roots of pomelo (Citrus maxima) in Thailand as Phytophthora nicotianae by the internal transcribed spacer ribosomal DNA sequence analysis. Then, we examined the in vitro and in vivo effects of Chaetomium globosum, Chaetomium lucknowense, Chaetomium cupreum and their crude extracts as biological control agents in controlling this P. nicotianae strain. Represent as antagonists in biculture test, the tested Chaetomium species inhibited mycelial growth by 50~56% and parasitized the hyphae, resulting in degradation of P. nicotianae mycelia after 30 days. The crude extracts of these Chaetomium species exhibited antifungal activities against mycelial growth of P. nicotianae, with effective doses of 2.6~101.4 µg/mL. Under greenhouse conditions, application of spores and methanol extracts of these Chaetomium species to pomelo seedlings inoculated with P. nicotianae reduced root rot by 66~71% and increased plant weight by 72~85% compared to that in the control. The method of application of antagonistic spores to control the disease was simple and economical, and it may thus be applicable for large-scale, highly effective biological control of this pathogen.

  17. Production and Optimization of Exo and Endocellulases from Thermophilic Fungi Scytalidium thermophilum SKESMBKU02

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    Sriyamjalla SANTOSHKUMAR

    2016-01-01

    Full Text Available The study aimed at isolation and screening of thermophilic cellulase producer, optimization and stability studies for maximum cellulase production. Fifteen thermophilic cellulase producing fungi were isolated from thermogenic habitats (vegetable market compost, mushroom compost, horse dung, municipal waste, nests of birds, decomposing litter, soils from furnace area, cattle dung, zoo dump, and industrial waste. of Telangana. All the fungal isolates were screened for their ability to produce cellulases. Scytalidium thermophilum SKESMBKU02 showed the highest cellulase activity in screening and was selected for further studies. The results showed that S. thermophilum SKESMBKU02 found to have high cellulolytic activity at 45 °C and pH 5.0 - 6.0. Optimization of enzyme production was studied in different carbon and nitrogen sources. The endo and exoglucanase activities were higher in media containing glucose as their carbon source followed by xylose. Yeast extract and peptone were good nitrogen sources for endoglucanase and exoglucanase activity respectively. The organism showed maximum dry weight in (NH42SO4 and NH4Cl. The exo and endocellulases produced by the S. thermophilum SKESMBKU02 were highly stable at pH 8.0 and temperature of 75 °C. The results indicate that the endo and exocellulases produced by the fungi were more stable at high temperature and alkaline pH.

  18. Potentially harmful secondary metabolites produced by indoor Chaetomium species on artificially and naturally contaminated building materials

    DEFF Research Database (Denmark)

    Dosen, Ina; Nielsen, Kristian Fog; Clausen, Geo;

    2017-01-01

    The presence of the fungal genus Chaetomium and its secondary metabolites in indoor environments is suspected to have a negative impact on human health and wellbeing. About 200 metabolites have been currently described from Chaetomium spp., but only the bioactive compound group, chaetoglobosins, ...

  19. Bioassays guided isolation of compounds from Chaetomium globosum.

    Science.gov (United States)

    Awad, N E; Kassem, H A; Hamed, M A; El-Naggar, M A A; El-Feky, A M M

    2014-06-01

    The aim of the present study was to evaluate different biological activities of the fungus Chaetomium globosum (family Chaetomiaceae). The evaluation was done through testing its antimicrobial, antioxidant and anticancer effects. C. globosum was isolated from the Cucumber soil (rhizosphere) and caused inhibition of the mycelial growth of Fusarium solani, Rhizoctonia solani and Sclerotium rolfsii in the biculture test. Petroleum ether and ethyl acetate extracts of the liquid culture of C. globosum showed potent in vitro antioxidant activity. C. globosum proved potent antibacterial activity against Bacillus subtilis, Escherichia coli and Pseudomonas fluorescens. It also recorded significant antifungal activity against Candida albicans, F. solani, Fusarium oxysporum, R. solani and Pythium ultimum. It exerted cytotoxic effect on human hepatocellular carcinoma cell line (HepG2). Unsaponifiable and saponifiable matters of the petroleum ether extract showed the presence of hydrocarbons, sterols and fatty acids. The ethyl acetate extract showed the presence of prenisatin, chrysophanol, chrysazin, chaetoviridin A and B. The isolated secondary metabolites proved significant antioxidant and antimicrobial activity on B. subtilis, E. coli and R. solani. In conclusion, this fungus showed different biological activities. Further studies must be done to apply its use in the agricultural and medicinal field.

  20. 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...... of its unusual habitat, which is a geothermal area with ground temperatures of 55-65 °C. Affinities clearly lie with the annual H. hirsutissimum Grauer, which is distributed in N Africa, the East Mediterranean area, and SW Asia; the latter, however, is hexaploid (2n = 48) and never occurs in thermal...

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

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    Juturu, Veeresh; Wu, Jin Chuan

    2012-01-01

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

  2. Potentially harmful secondary metabolites produced by indoor Chaetomium species on artificially and naturally contaminated building materials

    DEFF Research Database (Denmark)

    Dosen, Ina; Nielsen, Kristian Fog; Clausen, Geo

    2017-01-01

    , have been screened for, and thus detected in buildings. In this study, we used a liquid chromatography-high resolution mass spectrometry approach to screen both artificially and naturally infected building materials for all the Chaetomium metabolites described in the literature. Pure agar cultures were...

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

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    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.

  4. Invasive pulmonary mycosis due to Chaetomium globosum with false-positive galactomannan test: a case report and literature review.

    Science.gov (United States)

    Capoor, Malini R; Agarwal, Poojan; Goel, Manoj; Jain, Sarika; Shivaprakash, Mandya Rudramurthy; Honnavar, Prasanna; Gupta, Sunita; Chakrabarti, Arunaloke

    2016-03-01

    In this case, the authors report Chaetomium globosum as a cause of invasive pulmonary infection in a patient with Wegener's granulomatosis. Fungal hyphae (KOH and Calcofluor) were seen on direct microscopy of lung biopsy sample and bronchoalveolar lavage (BAL) sample. C. globosum isolated on culture clinched the diagnosis of invasive pulmonary infection by Chaetomium spp. A positive galactomannan of serum and BAL was repeatedly seen and was utilised for follow-up and as prognostic marker in patient management. The patient was successfully treated with liposomal amphotericin B followed by voriconazole. All the Chaetomium infections reported till date since 1980 are reviewed. Chaetomium spp. with its unique ecology has a hidden clinical potential to cause invasive mould infections.

  5. Producción y caracterización de la fenol oxidasa de Scytalidium thermophilum

    OpenAIRE

    Yasmin Sánchez-Rosario; José E. Sánchez; Rafael Vázquez-Duhalt; René H Andrade-Gallegos

    2011-01-01

    En este trabajo se estudió la producción de fenol oxidasa de tres cepas de Scytalidium thermophilum y se caracterizó la enzima de la cepa más productora. Se evaluó la producción de enzima en tres diferentes medios de cultivo: almidón (SM), papa dextrosa y levadura (PDY) e infusión de pasto (GI) a diferentes temperaturas (42, 45 y 48 °C) y diferentes valores de pH (6, 7 y 8). La actividad enzimática más alta (0.115 U/ml) fue observada con la cepa ECS-0602 en infusión de pasto Pangola a pH 8 y ...

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

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    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.

  7. Method for rapid detection and identification of chaetomium and evaluation of resistance to peracetic acid.

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    Nakayama, Motokazu; Hosoya, Kouichi; Tomiyama, Daisuke; Tsugukuni, Takashi; Matsuzawa, Tetsuhiro; Imanishi, Yumi; Yaguchi, Takashi

    2013-06-01

    In the beverage industry, peracetic acid has been increasingly used as a disinfectant for the filling machinery and environment due to merits of leaving no residue, it is safe for humans, and its antiseptic effect against fungi and endospores of bacteria. Recently, Chaetomium globosum and Chaetomium funicola were reported resistant to peracetic acid; however, little is known concerning the detail of peracetic acid resistance. Therefore, we assessed the peracetic acid resistance of the species of Chaetomium and related genera under identical conditions and made a thorough observation of the microstructure of their ascospores by transmission electron microscopy. The results of analyses revealed that C. globosum and C. funicola showed the high resistance to peracetic acid (a 1-D antiseptic effect after 900 s and 3-D antiseptic effect after 900 s) and had thick cell walls of ascospores that can impede the action mechanism of peracetic acid. We also developed specific primers to detect the C. globosum clade and identify C. funicola by using PCR to amplify the β-tubulin gene. PCR with the primer sets designed for C. globosum (Chae 4F/4R) and C. funicola (Cfu 2F/2R) amplified PCR products specific for the C. globosum clade and C. funicola, respectively. PCR with these two primer sets did not detect other fungi involved in food spoilage and environmental contamination. This detection and identification method is rapid and simple, with extremely high specificity.

  8. Indole diketopiperazines from endophytic Chaetomium sp 88194 induce breast cancer cell apoptotic death.

    Science.gov (United States)

    Wang, Fu-qian; Tong, Qing-yi; Ma, Hao-ran; Xu, Hong-feng; Hu, Song; Ma, Wei; Xue, Yong-bo; Liu, Jun-jun; Wang, Jian-ping; Song, Hong-ping; Zhang, Jin-wen; Zhang, Geng; Zhang, Yong-hui

    2015-03-19

    Diketopiperazines are important secondary metabolites of the fungi with variety bioactivities. Several species belonging to genus Chaetomium produce compounds of this class, such as chetomin. To identify new antitumor agents, secondary metabolites of fungus Chaetomium sp 88194 were investigated and three new indole diketopiperazines, Chaetocochins G (1), Oidioperazines E (2) and Chetoseminudin E (3), along with two known compounds Chetoseminudins C (4) and N-acetyl-β-oxotryptamine (5), were obtained. Chaetocochins G and Chetoseminudin E were recrystallized in CHCl3 containing a small amount of MeOH, and their structures with absolute configuration were established by spectroscopic data interpretation and single-crystal X-ray diffraction analysis. The absolute configuration of Oidioperazines E was defined by comparing of experimental and calculated electronic circular dichroism spectra. These isolates were also evaluated the anticancer activity, and Chaetocochins G displayed more potent cytotoxicity in MCF-7 cells than the common chemotherapeutic agent (5-fluorouracil) associated with G2/M cell cycle arrest. More importantly, Chaetocochins G induced cell apoptotic death via caspase-3 induction and proteolytic cleavage of poly (ADP-ribose) polymerase, concomitantly with increased Bax and decreased Bcl-2 expression. Our findings suggested that indole diketopiperazines from endophytic Chaetomium sp 88194 may be potential resource for developing anti-cancer reagents.

  9. Chaetoglobosins produced by Chaetomium globosum, endophytic fungus found in association with Viguiera robusta Gardn (Asteraceae); Chaetoglobosinas produzidas por Chaetomium globosum, fungo endofitico associado a Viguiera robusta Gardn. (Aasteraceae)

    Energy Technology Data Exchange (ETDEWEB)

    Momesso, Luciano da S.; Kawano, Cristina Y.; Ribeiro, Patricia H.; Nomizo, Auro; Goldman, Gustavo H.; Pupo, Monica T. [Universidade de Sao Paulo (USP), Ribeirao Preto, SP (Brazil). Faculdade de Ciencias Farmaceuticas]. E-mail: mtpupo@fcfrp.usp.br

    2008-07-01

    Endophytes live in association with host plants during all or part of their life cycle without causing any apparent disease. They are considered outstanding and underexploited sources of novel bioactive compounds. Chaetomium globosum was isolated as an endophytic fungus from the healthy leaves of Viguiera robusta. C. globosum is a remarkable producer of chaetoglobosins, which are typically cytotoxic. In this work, chaetoglobosins B (1), D (2) and E (3) have been produced by the endophytic C. globosum strain. Chaetoglobosin B was evaluated against Jurkat (leukemia) and B16F10 (melanoma) tumoral cells and showed 89.55% and 57.10% of inhibition at 0.1 mg mL{sup -1}, respectively. Chaetoglobosin B also showed weak antibacterial activity against Staphylococcus aureus (MIC 120 {mu}g/mL) and Escherichia coli (MIC 189 {mu}g/mL). (author)

  10. 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

    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...

  11. Isolamento de Chaetomium spp. em lesões subcutâneas de um cão: relato de caso Isolation of Chaetomium spp. on subcutaneous lesions of a dog: case report

    Directory of Open Access Journals (Sweden)

    C.A.S.B. Braga

    2013-02-01

    Full Text Available Caracterizou-se clinicamente a infecção pelo Chaetomium spp. em um cão, e descreveu-se seu isolamento e identificação. Ao exame dermatológico foram observadas pápulas nas orelhas, no tronco lateral e nos membros pélvicos. Ao rompimento de uma dessas pápulas, fluiu um líquido serosanguinolento com consequente úlcera no local. Foi colhido material para isolamento micológico, por meio de raspado das pápulas da orelha e da cauda. O diagnóstico foi micose subcutânea por Chaetomium spp.This work aimed to clinically characterize the infection by Chaetomium spp. in a dog, as well as describe its isolation and identification. Upon dermatological exam, papules on ears, lateral trunk and pelvic members were noticed. After the disruption of these papules there was serosanguineous secretion flowed by consequent ulcer in the region. Material for mycological isolation was picked, and a scraping of papules from ear and tail was done. The diagnosis was subcutaneous mycosis caused by Chaetomium spp.

  12. Purification and properties of thermostable tryptophanase from an obligately symbiotic thermophile, Symbiobacterium thermophilum.

    Science.gov (United States)

    Suzuki, S; Hirahara, T; Horinouchi, S; Beppu, T

    1991-12-01

    A thermostable tryptophanase was extracted from a thermophilic bacterium, Symbiobacterium thermophilum strain T, which is obligately symbiotic with the thermophilic Bacillus strain S. The enzyme was purified 21-fold to homogeneity with 19% recovery by a series of chromatographies using anion-exchange, hydroxylapatite, hydrophobic interaction, and MonoQ anion-exchange columns. The molecular weight of the purified enzyme was estimated to be approximately 210,000 by gel filtration, while the molecular weight of its subunit was 46,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which indicates that the native enzyme is composed of four homologous subunits. The isoelectric point of the enzyme was 4.9. The tryptophanase was stable to heating at 65 degrees C for 20 min and the optimum temperature for the enzyme activity for 20 min reaction was 70 degrees C. The optimum pH was 7.0. The NH2-terminal amino acid sequence of this tryptophanase shows similarity to that of Escherichia coli K-12, despite a great difference in the thermostability of these two enzymes. The purified enzyme catalyzed the degradation (alpha, beta-elimination) of L-tryptophan into indole, pyruvate, and ammonia in the presence of pyridoxal-5'-phosphate. The Km value for L-tryptophan was 1.47 mM. 5-Hydroxy-L-tryptophan, 5-methyl-DL-tryptophan, L-cysteine, S-methyl-L-cysteine, and L-serine were also used as substrates and converted to pyruvate. The reverse reaction of alpha, beta-elimination of this tryptophanase produced L-tryptophan from indole and pyruvate in the presence of a high concentration of ammonium acetate.

  13. 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.

    , among which Chaetomium sp. (NIOCC 36) was found to grow in a wide range of pH (between 4 and 12). This alkaline tolerant fungus was further tested for production of alkaline cellulases (Beta-endoglucanase, Beta-exoglucanase, Beta-glucosidase) using...

  14. Cellulase-free xylanases from Bacillus and other microorganisms.

    Science.gov (United States)

    Subramaniyan, S; Prema, P

    2000-02-01

    Xylanases are used mainly in the pulp and paper industries for the pretreatment of Kraft pulp prior to bleaching to minimize use of chlorine, the conventional bleaching agent. This application has great potential as an environmentally safe method. Hydrolysis by xylanases of relocated and reprecipitated xylan on the surface of cellulose fibres formed during Kraft cooking facilitates the removal of lignin by increasing permeability to oxidising agents. Most of the xylanases reported in the literature contained significant cellulolytic activity, which make them less suitable for pulp and paper industries. The need for large quantities of xylanases which would be stable at higher temperatures and pH values and free of cellulase activity has necessitated a search for novel enzymes. We have isolated and characterised several xylanase-producing cultures, one of which (an alkalophilic Bacillus SSP-34) produced more than 100 IU ml(-1) of xylanase activity. The SSP-34 xylanases have optimum activity at 50 degrees C in a pH range 6-8, with only small amounts of cellulolytic activity (CMCase (0.4 IU ml(-1), pH 7), FPase (0.2 IU ml(-1), pH 7) and no activity at pH 9).

  15. Cytoglobosins H and I, New Antiproliferative Cytochalasans from Deep-Sea-Derived Fungus Chaetomium globosum

    Directory of Open Access Journals (Sweden)

    Zhihan Zhang

    2016-12-01

    Full Text Available Cytoglobosins H (1 and I (2, together with seven known cytochalasan alkaloids (3–9, were isolated from the deep-sea-derived fungus Chaetomium globosum. The structures of new compounds 1 and 2 were elucidated by extensive 1D and 2D NMR and mass spectroscopic data. All the compounds were evaluated for their antiproliferative activities against MDA-MB-231 human breast cancer cells, LNCaP human prostate cancer cells, and B16F10 mouse melanoma cells. Compound 6 showed significant antiproliferative activity against LNCaP and B16F10 cell lines with IC50 values of 0.62 and 2.78 μM, respectively. Further testing confirmed that compound 6 inhibited the growth of LNCaP cells by inducing apoptosis.

  16. Cytoglobosins H and I, New Antiproliferative Cytochalasans from Deep-Sea-Derived Fungus Chaetomium globosum

    Science.gov (United States)

    Zhang, Zhihan; Min, Xitian; Huang, Junjun; Zhong, Yue; Wu, Yuehua; Li, Xiaoxia; Deng, Yinyue; Jiang, Zide; Shao, Zongze; Zhang, Lianhui; He, Fei

    2016-01-01

    Cytoglobosins H (1) and I (2), together with seven known cytochalasan alkaloids (3–9), were isolated from the deep-sea-derived fungus Chaetomium globosum. The structures of new compounds 1 and 2 were elucidated by extensive 1D and 2D NMR and mass spectroscopic data. All the compounds were evaluated for their antiproliferative activities against MDA-MB-231 human breast cancer cells, LNCaP human prostate cancer cells, and B16F10 mouse melanoma cells. Compound 6 showed significant antiproliferative activity against LNCaP and B16F10 cell lines with IC50 values of 0.62 and 2.78 μM, respectively. Further testing confirmed that compound 6 inhibited the growth of LNCaP cells by inducing apoptosis. PMID:27999388

  17. 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.

  18. XYLANASE PREBLEACHING ON NAOH-AQ WHEAT STRAW PULP

    Institute of Scientific and Technical Information of China (English)

    Caixia Li; Yongjun Deng; Ping Li; Guigan Fang; Shuchai Liu

    2004-01-01

    Before calcium hypochlorite bleaching (H) and chlorination,alkaline extraction, calcium hypochlorite three-stage-bleaching (CEH),we used a kind of hemicellulase, xylanase, to treat wheat straw pulp from Gaoyou Papermill.Xylanase pretreatment contained tow stages, the first stage was xylanase treatment, which was followed by alkaline extraction, the second stage. The xylanase could act on partial lignin and carbohydrate, chiefly xylan. The following alkaline extraction could dissolve something that could not be removed during the first stage. The result of pretreatment was to facilitate penetration of bleaching chemicals, to reduce effective chlorine consumption and to lower pollution loading of bleaching effluent. In the case of these two bleaching processes, the enzymatic pretreatment substantially enhanced the optical properties of the pulps. To calcium hypochlorite bleaching, strength properties of pulps were improved.

  19. XYLANASE PREBLEACHING ON NaOH-AQ WHEAT STRAW PULP

    Institute of Scientific and Technical Information of China (English)

    CaixiaLi; YongjunDeng; PingLi; GuiganFang; ShuchaiLiu

    2004-01-01

    Before calcium hypochlorite bleaching (H) and chlorination, alkaline extraction, calcium hypochlorite three-stage-bleaching (CEH),we used a kind of hemicellulase, xylanase, to treat wheat straw pulpfrom Gaoyou Papermill. Xylanase pretreatment contained tow stages, the first stage was xylanase treatment, which was followed by alkaline extraction, the second stage. The xylanase could act on partial lignin and carbohydrate, chiefly xylan. The following alkaline extraction could dissolve something that could not be removed during the first stage. The result of pretreatment was to facilitate penetration of bleaching chemicals, to reduce effective chlorine consumption and to lower pollution loading of bleaching effluent. In the case of these two bleaching processes, the enzymatic pretreatment substantially enhanced the optical properties of the pulps. To calcium hypochlorite bleaching, strength properties of pulps wereimproved.

  20. Xylanase supplementation to rye diets for growing pigs

    DEFF Research Database (Denmark)

    Nørgaard, Jan Værum; Pedersen, Trine Friis; Blaabjerg, Karoline;

    2016-01-01

    Supplementation of xylanase to pig diets can hydrolyze arabinoxylan (AX) into lower molecular weight compounds and thereby decrease the viscosity and improve nutrient utilization. Xylanase supplementation has, however, shown variable effects in diets containing wheat, rye, and combinations thereof....... Differences in animal age, enzyme source and dose, target substrate, and diet processing may explain this. The objective was to study the effect of increasing doses of endo-1,4-β-xylanase from Trichoderma reesei on apparent total tract digestibility (ATTD) of OM, starch, nonstarch polysaccharides (NSP), AX......, fat, N, and P of rye fed to growing pigs. Twenty-four 47-kg pigs were assigned to 4 diets containing 97.85% rye and 0, 4,000, 8,000, or 16,000 units xylanase/kg (as-fed basis). Pigs were placed in metabolic cages for 10 d: 5 d for adaption and 5 d for total but separate collection of feces and urine...

  1. Identification problems with sterile fungi, illustrated by a keratitis due to a non-sporulating Chaetomium-like species.

    Science.gov (United States)

    Vinod Mootha, V; Shahinpoor, P; Sutton, Deanna A; Xin, Lian; Najafzadeh, M J; de Hoog, G Sybren

    2012-05-01

    A 39-year-old farm worker was injured in her right eye by a piece of wire, which resulted in a corneal ulcer unresponsive to antibiotic treatment. The clinical appearance was that of a corneal infiltrate with feathery borders resembling fungal keratitis. Corneal scrapings were collected and the patient was started on natamycin 5% eye drops, fluconazole 0.3% eye drops, and oral fluconazole. A non-sporulating fungus was isolated from the samples. Based upon macroscopic and microscopic morphologic features, it was provisionally identified as a Papulaspora species due to the fact that members of this genus generally do not form diagnostically useful conidia. However, it was found through the use of ITS sequencing that the isolate clustered within the ascomycete genus Chaetomium. The sequence did not fully match with any sequences of available ex-type strains of Chaetomium, Thielavia and Papulaspora and hence might belong to an undescribed specie. However, without diagnostic morphological features the taxon cannot be introduced as a novel member of the genus Chaetomium. Antifungal susceptibility testing was performed according to published standards. The corneal ulcer was successfully treated with six weeks of antifungal therapy.

  2. Rapid development of xylanase assay conditions using Taguchi methodology.

    Science.gov (United States)

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

    2016-11-01

    The present investigation is mainly concerned with the rapid development of extracellular xylanase assay conditions by using Taguchi methodology. The extracellular xylanase was produced from Aspergillus niger (KP874102.1), a new strain isolated from a soil sample of the Baramura forest, Tripura West, India. Four physical parameters including temperature, pH, buffer concentration and incubation time were considered as key factors for xylanase activity and were optimized using Taguchi robust design methodology for enhanced xylanase activity. The main effect, interaction effects and optimal levels of the process factors were determined using signal-to-noise (S/N) ratio. The Taguchi method recommends the use of S/N ratio to measure quality characteristics. Based on analysis of the S/N ratio, optimal levels of the process factors were determined. Analysis of variance (ANOVA) was performed to evaluate statistically significant process factors. ANOVA results showed that temperature contributed the maximum impact (62.58%) on xylanase activity, followed by pH (22.69%), buffer concentration (9.55%) and incubation time (5.16%). Predicted results showed that enhanced xylanase activity (81.47%) can be achieved with pH 2, temperature 50°C, buffer concentration 50 Mm and incubation time 10 min.

  3. 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

  4. Cloning of xylanase genes from Ruminococcus albus and chromosome mapping of Fibrobacter succinogenes

    DEFF Research Database (Denmark)

    Nagamine, T.; Aminov, Rustam; Ogata, K.

    1997-01-01

    The characterization of the xylanase genes of Ruminococcus albus and the genomic organization of Fibrobacter succinogenes are described.......The characterization of the xylanase genes of Ruminococcus albus and the genomic organization of Fibrobacter succinogenes are described....

  5. 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.

    Thermoalkalophilic Enterobacter sp MTCC 5112 was isolated from a sediment sample collected from the Mandovi estuary, west coat of India. This culture produced extracellular xylanase. The xylanase enzyme was isolated by ammonium sulfate (80...

  6. 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

  7. Bioactive metabolites from Chaetomium globosum L18, an endophytic fungus in the medicinal plant Curcuma wenyujin.

    Science.gov (United States)

    Wang, Yanhong; Xu, Lei; Ren, Weiming; Zhao, Dan; Zhu, Yanping; Wu, Xiaomin

    2012-02-15

    An endophytic fungus, strain L18, isolated from the medicinal plant Curcuma wenyujin Y.H. Chen et C. Ling was identified as Chaetomium globosum Kunze based on morphological characteristics and sequence data for the internal transcribed spacer (ITS-5.8S-ITS2) of the nuclear ribosomal DNA. A new metabolite named chaetoglobosin X (1), together with three known compounds erogosterol (2), ergosterol 5α,8-peroside (3) and 2-methyl-3-hydroxy indole (4), were isolated from C. globosum L18. Their structures were elucidated by spectroscopic methods including NMR, UV, IR and MS data and comparison with published data. Chaetoglobosin X (1) is hitherto unknown, whereas 2-methyl-3-hydroxy indole (4) is reported for the first time as a fungal metabolite, and erogosterol (2) and ergosterol 5α,8-peroside (3) are known fungal metabolites previously identified in other genera. Chaetoglobosin X (1) exhibited a broader antifungal spectrum and showed the strongest cytotoxic activity against H22 and MFC cancer cell lines.

  8. 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.

  9. ENZYMATIC DEINKING OF OLD NEWSPRINT WITH CELLULASES AND XYLANASE

    Institute of Scientific and Technical Information of China (English)

    Shoujuan Wang; Menghua Qin; Yingjuan Fu; Zhiyong Shao

    2004-01-01

    The enzymatic deinking and fiber modification of old newsprint (ONP) with several cellulases and xylanase were investigated and the suitable enzyme candidates were selected for ONP deinking in this paper. The results demonstrated that the cellulases and hemicellulases could significantly improve the deinking efficiency and fiber modification.Moreover, the synergistic effects of Novozym342 and xylanase (HC) can further enhance the deinking performance, reduce the dirt count and improve the brightness of resulting pulp. Additionally, compared to deinked pulps, obtained from conventional chemical materials, enzymatically deinked pulps had better bleachability, and the brightness of the bleached pulp reached 59.1% ISO, 9% ISO higher than the unbleached pulp.

  10. ENZYMATIC DEINKING OF OLD NEWSPRINT WITH CELLULASES AND XYLANASE

    Institute of Scientific and Technical Information of China (English)

    ShoujuanWang; MenghuaQin; YingjuanFu; ZhiyongShao

    2004-01-01

    The enzymatic deinking and fiber modification of old newsprint (ONP) with several cellulases and xylanase were investigated and the suitable enzyme candidates were selected for ONP deinking in this paper. The results demonstrated that the cellulases and hemicellulases could significantly improve the deinking efficiency and fiber modification. Moreover, the synergistic eflbcts of Novozym342 and xylanase (HC) can further enhance the deinking performance, reduce the dirt count and improve the brightness of resulting pulp. Additionally, compared to deinked pulps, obtained from conventional chemical materials, enzymatically deinked pulps had better bleachability, and the brightness of the bleached pulp reached 59.1% ISO, 9% ISO higher than the unbleached pulp.

  11. 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.

  12. Cannibalism as an interacting phenotype: precannibalistic aggression is influenced by social partners in the endangered Socorro Isopod (Thermosphaeroma thermophilum).

    Science.gov (United States)

    Bleakley, B H; Welter, S M; McCauley-Cole, K; Shuster, S M; Moore, A J

    2013-04-01

    Models for the evolution of cannibalism highlight the importance of asymmetries between individuals in initiating cannibalistic attacks. Studies may include measures of body size but typically group individuals into size/age classes or compare populations. Such broad comparisons may obscure the details of interactions that ultimately determine how socially contingent characteristics evolve. We propose that understanding cannibalism is facilitated by using an interacting phenotypes perspective that includes the influences of the phenotype of a social partner on the behaviour of a focal individual and focuses on variation in individual pairwise interactions. We investigated how relative body size, a composite trait between a focal individual and its social partner, and the sex of the partners influenced precannibalistic aggression in the endangered Socorro isopod, Thermosphaeroma thermophilum. We also investigated whether differences in mating interest among males and females influenced cannibalism in mixed sex pairs. We studied these questions in three populations that differ markedly in range of body size and opportunities for interactions among individuals. We found that relative body size influences the probability of and latency to attack. We observed differences in the likelihood of and latency to attack based on both an individual's sex and the sex of its partner but found no evidence of sexual conflict. The instigation of precannibalistic aggression in these isopods is therefore a property of both an individual and its social partner. Our results suggest that interacting phenotype models would be improved by incorporating a new conditional ψ, which describes the strength of a social partner's influence on focal behaviour.

  13. Biological evaluation of endophytic fungus, Chaetomium globosum JN711454, as potential candidate for improving drug discovery.

    Science.gov (United States)

    Selim, Khaled A; El-Beih, Ahmed A; Abdel-Rahman, Tahany M; El-Diwany, Ahmed I

    2014-01-01

    The main objective of this research work focused on investigating the biological and chemical aspects of endophytic fungus Chaetomium globosum, for pharmaceutical purposes to improve the drug discovery process. The endophytic C. globosum was isolated from healthy leaves of Egyptian medicinal plant Adiantum capillus-veneris collected from Saint Katherine Protectorate, Sinai, Egypt. The identification of C. globosum was on the basis of classical and molecular taxonomy. Gene encoding for 18S rRNA was partially sequenced, submitted to the GenBank and got the accession number JN711454, to resolve the phylogenetic relations with fungal ancestor using phylogenetic tree. To explore the biosynthetic power of endophytic C. globosum JN711454, the fungus was cultivated over five different media, oatmeal, rice, yeast malt glucose, potato dextrose agar (PDA) and Czapek's dox media, for 3 weeks at 30 °C, followed by extraction with different solvents, ethyl acetate (EA), and methanol. The ethyl acetate extract of C. globosum cultivated on PDA medium was the most potent extract. It showed strong antioxidant activity with EC50 11.5 μg/ml, potent anticancer activity with 55 % toxicity toward HepG-2 cells at 100 μg/ml and 66 % cytotoxicity to FGC4 cells at 250 μg/ml, promising butyrylcholinesterase inhibitory activities (>85 %), and moderate antimicrobial and stopped the attachment of HSV-2 virus to VERO cells. The metabolomic profiling of PDA-EA extract using LC-MS revealed the presence of several metabolites to which the observed bioactivities could be attributed. Here we report for the first time inhibitory activity of endophytic C. globosum JN711454 secondary metabolites to butyrylcholinesterase, one of neuro hydrolase enzymes that play a major role in development of Alzheimer's disease.

  14. 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...

  15. Visualization of the structural changes in plywood and gypsum board during the growth of Chaetomium globosum and Stachybotrys chartarum

    DEFF Research Database (Denmark)

    Lewinska, Anna Malgorzata; Hoof, Jakob Blæsbjerg; Peuhkuri, Ruut H.

    2016-01-01

    Fungal growth in indoor environments is associated with many negative health effects. Many studies focus on brown- and white-rot fungi and their effect on wood, but there is none that reveals the influence of soft-rot fungi, such as Stachybotrys spp. and Chaetomium spp., on the structure of build...

  16. 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.

  17. Polyethyleneimine as a promoter layer for the immobilization of cellobiose dehydrogenase from Myriococcum thermophilum on graphite electrodes.

    Science.gov (United States)

    Schulz, Christopher; Ludwig, Roland; Gorton, Lo

    2014-05-01

    Cellobiose dehydrogenase (CDH) is a promising enzyme for the construction of biofuel cell anodes and biosensors capable of oxidizing aldoses as cellobiose as well as lactose and glucose and with the ability to connect to an electrode through a direct electron transfer mechanism. In the present study, we point out the beneficial effect of a premodification of spectrographic graphite electrodes with the polycation polyethyleneimine (PEI) prior to adsorption of CDH from Myriococcum thermophilum (MtCDH). The application of PEI shifts the pH optimum of the response of the MtCDH modified electrode from pH 5.5 to 8. The catalytic currents to lactose were increased up to 140 times, and the K(M)(app) values were increased up to 9 times. The previously investigated, beneficial effect of divalent cations on the activity of CDH was also present for graphite/PEI/MtCDH electrodes but was less pronounced. Polarization curves revealed a second unexpected catalytic wave for graphite/PEI/MtCDH electrodes especially pronounced at pH 8. Square wave voltammetric studies revealed the presence of an unknown redox functionality present at 192 mV vs Ag|AgCl (0.1 M KCl) at pH 8, probably originating from an oxidized adenosine derivative. Adenosine is a structural part of the flavin adenine dinucleotide (FAD) cofactor of the dehydrogenase domain of CDH. It is suggested that for some enzyme molecules FAD leaks out from the active site, adsorbs onto graphite, and is oxidized on the electrode surface into a product able to mediate the electron transfer between CDH and the electrode. PEI is suggested and discussed to act in several manners by (a) increasing the surface loading of the enzyme, (b) possibly increasing the electron transfer rate between CDH and the electrode, and (c) facilitating the creation or immobilization of redox active adenosine derivatives able to additionally mediate the electron transfer between CDH and the electrode.

  18. Production, properties and application to biocatalysis of a novel extracellular alkaline phenol oxidase from the thermophilic fungus Scytalidium thermophilum.

    Science.gov (United States)

    Ogel, Z B; Yüzügüllü, Y; Mete, S; Bakir, U; Kaptan, Y; Sutay, D; Demir, A S

    2006-08-01

    Scytalidium thermophilum produces an extracellular phenol oxidase on glucose-containing medium. Certain phenolic acids, specifically gallic acid and tannic acid, induce the expression of the enzyme. Production at 45 degrees C in batch cultures is growth-associated and is enhanced in the presence of 160 microM CuSO4 x 5 H2O and 3 mM gallic acid. The highest enzyme activity is observed at pH 7.5 and 65 degrees C, on catechol. When incubated for 1 h at pH 7 and pH 8, 95% and 86% of the activity is retained. Thermostability decreases gradually from 40 degrees C to 80 degrees C. Estimated molecular mass is c. 83 kDa, and pI is acidic at c. 5.4. Substrate specificity and inhibition analysis in culture supernatants suggest that the enzyme has unique properties showing activity towards catechol; 3,4-dihydroxy-L-phenylalanine (L-DOPA); 4-amino-N, N-diethylaniline (ADA); p-hydroquinone; gallic acid; tannic acid and caffeic acid, and no activity towards L-tyrosine, guaiacol, 2,2'-azino-bis(3-ethyl-benzthiazoline-6-sulphonic acid) (ABTS) and syringaldazine. Inhibition is observed in the presence of salicyl hydroxamic acid (SHAM) and p-coumaric acid. Enzyme activity is enhanced by cetyltrimethylammonium bromide (CTAB) and polyvinylpyrrolidone (PVP), and the organic solvents dimethyl sulfoxide (DMSO) and ethanol. No inhibition is observed in the presence of carbon monoxide. Benzoin, benzoyl benzoin and hydrobenzoin are converted into benzil, and stereoselective oxidation is observed on hydrobenzoin. The reported enzyme is novel due to its catalytic properties resembling mainly catechol oxidases, but displaying some features of laccases at the same time.

  19. 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.

  20. Chaetoglobosinas produzidas por Chaetomium globosum, fungo endofítico associado a Viguiera robusta Gardn. (Asteraceae Chaetoglobosins produced by Chaetomium globosum, endophytic fungus found in association with Viguiera robusta Gardn (Asteraceae

    Directory of Open Access Journals (Sweden)

    Luciano da S. Momesso

    2008-01-01

    Full Text Available Endophytes live in association with host plants during all or part of their life cycle without causing any apparent disease. They are considered outstanding and underexploited sources of novel bioactive compounds. Chaetomium globosum was isolated as an endophytic fungus from the healthy leaves of Viguiera robusta. C.globosum is a remarkable producer of chaetoglobosins, which are typically cytotoxic. In this work, chaetoglobosins B (1, D (2 and E (3 have been produced by the endophytic C. globosum strain. Chaetoglobosin B was evaluated against Jurkat (leukemia and B16F10 (melanoma tumoral cells and showed 89.55% and 57.10% of inhibition at 0.1 mg mL-1, respectively. Chaetoglobosin B also showed weak antibacterial activity against Staphylococcus aureus (MIC 120 µg/mL and Escherichia coli (MIC 189 µg/mL.

  1. Safety evaluation of a xylanase expressed in Bacillus subtilis.

    Science.gov (United States)

    Harbak, L; Thygesen, H V

    2002-01-01

    A programme of studies was conducted to establish the safety of a xylanase expressed in a self-cloned strain of Bacillus subtilis to be used as a processing aid in the baking industry. To assess acute and subchronic oral toxicity, rat feeding studies were conducted. In addition, the potential of the enzyme to cause mutagenicity and chromosomal aberrations was assessed in microbial and tissue culture in vitro studies. Acute and subchronic oral toxicity was not detected at the highest dose recommended by OECD guidelines. There was no evidence of mutagenic potential or chromosomal aberrations. Furthermore, the organism used for production of the xylanase is already accepted as safe by several major national regulatory agencies.

  2. Four new steroids from the endophytic fungus Chaetomium sp. M453 derived of Chinese herbal medicine Huperzia serrata.

    Science.gov (United States)

    Yu, Fei-Xue; Li, Zhe; Chen, Yao; Yang, Yin-He; Li, Guo-Hong; Zhao, Pei-Ji

    2017-03-01

    An endophytic fungus, Chaetomium sp. M453, was isolated from Huperzia serrata (Thunb. ex Murray) Trev. and subjected to phytochemical investigation. Three unusual C25 steroids, neocyclocitrinols E-G (1-3), and 3β-hydroxy-5,9-epoxy-(22E,24R)-ergosta-7,22-dien-6-one (4) together with three known steroids were isolated from solid fermentation products of the fungus, which were elucidated by extensive spectroscopic analyses, including 1D-, 2D-NMR, and HR-ESI-MS experiments. The absolute configuration of 1 was determined by X-ray crystallographic analysis and CD analyses. The acetylcholinesterase inhibitory activities of compounds 1-4 were tested in vitro. Compound 4 showed weak acetylcholinesterase inhibitory activity.

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

    NARCIS (Netherlands)

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

    2016-01-01

    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 af

  4. Bifunctional xylanases and their potential use in biotechnology

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Numan, M.Th.

    is half as sweet as sucrose, can be applicable to foods as a sweetener that is capable of improving diabetic symptoms [37]. Concluding remarks This review provides the information on most of the aspects of bifunctional enzyme with special reference... of the bifunctional xylanases it is necessary in future to utilize such hybrid protein as an alternative to expensive and polluting chemical treatments or to improve already existing enzymatic processes for utilization of veg- etal by-products in the agro...

  5. 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.

  6. 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.

    Thermoalkalophilic Arthrobacter sp. produced extracellular xylanase, when wheat bran, rice husk, rice bran and bagassae were used as carbon source under solid-state fermentation (SSF). The xylanase enzyme was isolated by ammonium sulfate (80...

  7. Baker's asthma due to the enzyme xylanase -- a new occupational allergen.

    Science.gov (United States)

    Baur, X; Sander, I; Posch, A; Raulf-Heimsoth, M

    1998-12-01

    The asthmatic baker showed IgE-mediated sensitization to xylanase of Aspergillus niger used as a baking additive. Inhalative challenge with approximately 0.5 microg of the enzyme resulted in an immediate-type asthmatic reaction. This case, as well as a preliminary screening of symptomatic bakers, shows that xylanase is a further relevant type I-sensitizer in the baking industry.

  8. 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

    induced (140%) when pre-incubated with 0.5 M NaCl for 4 h. The xylanase gene, xyn40, from marine bacterium B. subtilis cho40 was cloned, and expressed in Escherichia coli. The xylanase gene was 645 bp long and had a 215 amino acid ORF protein with a...

  9. A novel thermoalkaliphilic xylanase from Gordonia sp. is salt, solvent and surfactant tolerant.

    Science.gov (United States)

    Kashyap, Radhika; Monika; Subudhi, Enketeswara

    2014-12-01

    Two aerobic bacterial consortia namely Con T and Con R were developed by enrichment technique from termite gut and cow dung respectively, using xylan as a sole carbon source. Molecular characterization of Con R based on 16S rRNA sequence analysis showed the presence of Pannonibacter sp. R-3 and Pseudoxanthomas sp. R-5. On the other hand, Con T showed the presence of Pseudoxanthomas sp. T-5, Cellulosimicrobium sp. T-21, and Gordonia sp. T-30. Being the maximum xylanase producer among the five isolates and being a novel xylanase producing bacterial genus, Gordonia sp. T-30 was selected. Xylanase produced by Gordonia sp. T-30 showed optimum activity at 60 °C and pH 9. Xylanase was 95% stable for 120 min at pH 9.0 and 98% stable at 60 °C for 90 min. Xylanase activity was stimulated in the presence of organic solvents such as petroleum ether, acetone, diethyl ether, n-hexane, and benzene. Detergent like cetyltrimethylammonium bromide and presence of NaCl also accelerated the xylanase function. Comparative evaluation was studied between sterilized and non-sterilized solid fermentation to produce xylanase by Gordonia sp. T-30 using various agricultural residues as growth substrate in cost effective manner. Industrially important features endowed by this xylanase make it a very promising candidate for food, feed, and fuel industry.

  10. 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.

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

    Science.gov (United States)

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

    2015-09-01

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

  12. [Cellulase and xylanase activity of phytopathogenic and endophytic fungal strains of Alternaria alternata (Fr.) Keissler].

    Science.gov (United States)

    Kurchenko, I M; Sokolova, O V; Zhdanova, N M; Iarynchyn, A M; Iovenko, O M

    2008-01-01

    A comparative analysis of cellulase and xylanase activity of 25 fungal strains of phytopathogenic and endophytic Alternaria alternata had been realized for the first time using the qualitative reactions. The rate of their linear growth on the media with carboxymethylcellulose or xylane had been studied. The cellulase and xylanase activities clearly depended on the distinct strain. The absence of distinct dependence of cellulase and xylanase activities on the species and organs of host plants was demonstrated. The majority of investigated strains of A. alternata did not possess a cellulase activity or the latter was low, but as a whole the phytopathogenic strains were more active than endophytic ones. Xylanase activity was considerable for the fungal strains of all trophyc groups. It was shown that the level of xylanase activity cannot become a biochemical marker of the A. alternata isolate pathogenicity.

  13. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  15. Production, purification, and characterization of a major Penicillium glabrum xylanase using Brewer's spent grain as substrate.

    Science.gov (United States)

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

    2013-01-01

    In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn(2+) and the reducing agents β -mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg(2+), Zn(2+), and Cu(2+) 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.

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

    Directory of Open Access Journals (Sweden)

    Adriana Knob

    2013-01-01

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

  17. Synergistic effect and application of xylanases as accessory enzymes to enhance the hydrolysis of pretreated bagasse.

    Science.gov (United States)

    Gonçalves, Geisa A L; Takasugi, Yusaku; Jia, Lili; Mori, Yutaro; Noda, Shuhei; Tanaka, Tsutomu; Ichinose, Hirofumi; Kamiya, Noriho

    2015-05-01

    Recently, the new trend in the second-generation ethanol industry is to use mild pretreatments, in order to reduce costs and to keep higher content of hemicellulose in the biomass. Nevertheless, a high enzyme dosage is still required in the conversion of (hemi)cellulose. The interaction between cellulases and xylanases seems to be an effective alternative to reduce enzyme loading in the saccharification process. At first, to evaluate the synergism of xylanases on bagasse degradation, we have produced two xylanases from glycoside hydrolase family 10 (GH10) and three xylanases from glycoside hydrolase family 11 (GH11), from two thermophilic organisms, Thermobifida fusca and Clostridium thermocellum, and one mesophilic organism, Streptomyces lividans. Peracetic acid (PAA) pretreated bagasse was used as substrate. The combination of XynZ-C (GH10, from C. thermocellum), and XlnB (GH11, from S. lividans) presented the highest degree of synergy after 6h (3.62). However, the combination of XynZ-C and Xyn11A (GH11, from T. fusca) resulted in the highest total yield of reducing sugars. To evaluate the synergism between xylanases and cellulases, commercial cellulase preparation from Trichoderma reesei was combined with the selected xylanases, XynZ-C and Xyn11A. About 2-fold increase was observed in the concentration of reducing sugars, when both xylanases, XynZ-C and Xyn11A, were added together with T. reesei cellulases in the reaction mixture.

  18. Novel thermostable endo-xylanase cloned and expressed from bacterium Geobacillus sp. WSUCF1.

    Science.gov (United States)

    Bhalla, Aditya; Bischoff, Kenneth M; Uppugundla, Nirmal; Balan, Venkatesh; Sani, Rajesh K

    2014-08-01

    A gene encoding a GH10 endo-xylanase from Geobacillus sp. WSUCF1 was cloned and expressed in Escherichia coli. Recombinant endo-xylanase (37kDa) exhibited high specific activity of 461.0U/mg of protein. Endo-xylanase was optimally active on birchwood xylan at 70°C and pH 6.5. The endo-xylanase was found to be highly thermostable at 50 and 60°C, retaining 82% and 50% of its original activity, respectively, after 60h. High xylan conversions (92%) were obtained with oat-spelt xylan hydrolysis. Higher glucan and xylan conversions were obtained on AFEX-treated corn stover with an enzyme cocktail containing WSUCF1 endo-xylanase (71% and 47%) as compared to enzyme cocktail containing commercial fungal endo-xylanase (64% and 41%). High specific activity, active at high pH's, wide substrate specificity, and higher hydrolytic activity on recalcitrant lignocellulose, make this endo-xylanase a suitable candidate for biofuel and bioprocess industries.

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

    Science.gov (United States)

    Sakthiselvan, Punniavan; Naveena, Balakrishnan; Partha, Nagarajan

    2014-01-01

    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 7(th) 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.

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

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    Guo-Qiang Guan

    2016-01-01

    Full Text Available 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.

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

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    G. Ghoshal

    2016-12-01

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

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

    Directory of Open Access Journals (Sweden)

    Punniavan Sakthiselvan

    2014-12-01

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

  3. 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.

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

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    Aditya eBhalla

    2015-06-01

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

  5. Visualization of the structural changes in plywood and gypsum board during the growth of Chaetomium globosum and Stachybotrys chartarum.

    Science.gov (United States)

    Lewinska, Anna M; Hoof, Jakob B; Peuhkuri, Ruut H; Rode, Carsten; Lilje, Osu; Foley, Matthew; Trimby, Patrick; Andersen, Birgitte

    2016-10-01

    Fungal growth in indoor environments is associated with many negative health effects. Many studies focus on brown- and white-rot fungi and their effect on wood, but there is none that reveals the influence of soft-rot fungi, such as Stachybotrys spp. and Chaetomium spp., on the structure of building materials such as plywood and gypsum wallboard. This study focuses on using micro-computed tomography (microCT) to investigate changes of the structure of plywood and gypsum wallboard during fungal degradation by S. chartarum and C. globosum. Changes in the materials as a result of dampness and fungal growth were determined by measuring porosity and pore shape via microCT. The results show that the composition of the building material influenced the level of penetration by fungi as shown by scanning electron microscopy (SEM). Plywood appeared to be the most affected, with the penetration of moisture and fungi throughout the whole thickness of the sample. Conversely, fungi grew only on the top cardboard in the gypsum wallboard and they did not have significant influence on the gypsum wallboard structure. The majority of the observed changes in gypsum wallboard occurred due to moisture. This paper suggests that the mycelium distribution within building materials and the structural changes, caused by dampness and fungal growth, depend on the type of the material.

  6. Purification and characterization of a moderately thermostable xylanase from Bacillus sp. strain SPS-0.

    Science.gov (United States)

    Bataillon; Nunes Cardinali A; Castillon; Duchiron

    2000-02-01

    A Bacillus spp. strain SPS-0, isolated from a hot spring in Portugal, produced an extracellular xylanase upon growth on wheat bran arabinoxylan. The enzyme was purified to homogeneity by ammonium sulfate precipitation, anion exchange, gel filtration, and affinity chromatography. The optimum temperature and pH for activity was 75 degrees C and 6.0. Xylanase was stable up to 70 degrees C for 4 h at pH 6.0 in the presence of xylane. Xylanase was completely inhibited by the Hg(2+) ions. beta-Mercaptoethanol, dithiothreitol, and Mn(2+) stimulated the xylanase activity. The products of birchwood xylan hydrolysis were xylose, xylobiose, xylotriose, and xylotetraose. Kinetic experiments at 60 degrees C and pH 6.0 gave V(max) and K(m)values of 2420 nkat/mg and 0.7 mg/ml.

  7. [Cellulase and xylanase activities of Fusarium Lk:Fr. genus fungi of different trophic groups].

    Science.gov (United States)

    Kurchenko, I M; Sokolova, O V; Zhdanova, N M; Iarynchyn, A M; Iovenko, O M

    2008-01-01

    A comparative analysis of cellulase and xylanase activities of 26 fungal strains of phytopathogenic, saprophytic and endophytic Fusarium species has been realized using the qualitative reactions. The rare of their linear growth on the media with carboxymethyl cellulose or xylane has been studied. It was shown that the fungi of genus Fusarium belonging to different trophic groups possessed low activities of investigated enzymes as a whole, but in endophytic strains their levels were lower than in phytopathogenic ones. At the same time the distinct strain dependence of cellulase and xylanase activities was fixed in the fungi of different trophic groups. As far as the cellulase and xylanase activities in phytopathogenic isolates varied from complete absence to high levels, and since the activity maximum for each of the investigated strains was observed in different growth terms the conclusion was made that the cellulase and xylanase activities could not be considered as possible markers of the fungal isolate pathogenicity on the strain level.

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

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    Yasser Bakri

    2011-08-01

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

  9. Mutation-Screening in Xylanase-Producing Strains by Ion Implantation

    Institute of Scientific and Technical Information of China (English)

    李市场; 吴敏; 姚建铭; 潘仁瑞; 余增亮

    2005-01-01

    With ion implantation (N+, energy 10 keV and dosage 1.56×1015N+ cm-2), a high xylanase-producing strain Aspergillus niger N212 was selected. Based on an orthogonal experiment, an optimal fermentation condition was designed for this high-yield strain. The suitable medium was composed of 8% corncob; 1.0% wheat bran; 0.1% TWEEN20; 0.5% (NH4)2SO4;3.0×10-4. At present, under our experiment condition, xylanase activity of Aspergillus niger N212 reached a level of 600 IU/ml, almost increased by 100% in xylanase production and the time of yielding xylanase was largely reduced to 12 h at 28 ℃.

  10. Antimicrobial, antioxidant, and cytotoxic activities of endhopitic fungi Chaetomium sp. isolated from Phyllanthus niruri Linn: in vitro and in silico studies

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    Rollando Rollando

    2017-03-01

    Full Text Available Endophytic fungi Chaetomium sp isolated from Phyllanthus niruri Linn. Mycelium powder was extracted by using ethyl acetate. Extract was fractionated using n-hexane, dichloromethane and ethanol 96%. The antimicrobial test was carried out using disc diffusion and microdilution methods. The antioxidant activity of the fraction was determined using hydrogen peroxide free radical scavenging and reducing power capacity activities. The cytotoxicity assay of the fraction against T47D breast cancer cell was carried out using dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide method (MTT. The in silico prediction of chemical substances which are reported exist in Chaetomium sp. performed using AutoDockVina embedded in PyRx version 8.0. Dichloromethane fraction was found as the most active sample against Escherichia coli (IC50 20.76 mg/mL, Staphylococcus aureus (IC50 70.15 mg/mL, Salmonella typhi (49.13 mg/mL and was found as the most high phenolic content with value 47.44 mg GAE/g fraction, whereas the best antioxidant activity was performed by ethanol 96% fraction (85%. Cytotoxicity assay against T47D cell line showed dichloromethane fraction have highest activity with IC50 10.76 mg/mL. The docking studies showed that compounds bearing xanthone structure were potential for maltose binding periplasmic and human aromatase associating with their potencies as antibacteria and anticancer. Endophytic fungi Chaetomium sp. was isolated from Phyllanthus niruri using n-hexane, dichloromethane and ethanol fractions was studied its various biological activities as antimicrobial, antioxidant and cytotoxic agent against breast cancer cell.

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

    Science.gov (United States)

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

    2010-05-01

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

  12. Mutagenesis and Screening of High Yield Xylanase Production Strain of Aspergillus usamii by Microwave Irradiation

    Institute of Scientific and Technical Information of China (English)

    李永泉; 陈时飞; 岑沛霖

    2003-01-01

    A high yield xylanase producing strain, A. usamii L336-23, was screened out from its parent strain A.usamii L336 after microwave irradiation. Its productivity of xylanase activity increased by 35.7% from 21000μ·m1-1 to 28500μ·m1-1 and was stable after frequent subcultures and storage for more than two months.The mechanism of microwave mutation was also discussed.

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

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    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.

  14. Bacterial xylanase expression in mammalian cells and transgenic mice.

    Science.gov (United States)

    Fontes, C M; Ali, S; Gilbert, H J; Hazlewood, G P; Hirst, B H; Hall, J

    1999-06-11

    The energy which simple-stomached livestock can derive from dietary plant material is limited by the lack of plant polysaccharide degrading enzymes in their gastro-intestinal (GI) tract and the inefficient microbial fermentation of such material in their hind-gut. In poultry the non-starch polysaccharides found in cereal grains can also impair normal digestive function as they form viscous gels in the GI tract inhibiting the breakdown and absorption of nutrients. The nutrition of such livestock could, therefore, be improved by the introduction of enzymes able to degrade plant polysaccharides in the small intestine. We describe the expression of a xylanase, XYLY', from the bacterium Clostridium thermocellum in mammalian cells and the exocrine pancreas of transgenic mice. The enzyme is synthesised, secreted and functionally active in the eukaryote system. This work demonstrates the feasibility of generating animals with the endogenous capacity to depolymerise the xylan component of hemi-cellulose.

  15. Molecular breeding of transgenic rice expressing a xylanase domain of the xynA gene from Clostridium thermocellum.

    Science.gov (United States)

    Kimura, T; Mizutani, T; Tanaka, T; Koyama, T; Sakka, K; Ohmiya, K

    2003-09-01

    The gene encoding the catalytic domain of thermostable xylanase from Clostridium thermocellum F1 was expressed in rice plants under the control of a constitutive promoter. The gene encoding Xylanase A was modified to encode the catalytic domain of family 11 xylanase without the signal sequence (xynA1), and was introduced into rice plants and expressed under the control of a modified cauliflower mosaic virus 35S promoter. Zymogram analysis indicated that the recombinant xylanase was produced in rice plants. The xynA1 gene was stably expressed in rice straw and seed grains. No phenotypic effect of xylanase expression was noted. The enzyme was detected in the desiccated grain. High levels of enzyme activity were maintained in the cell-free extract during incubation at 60 degrees C for 24 h. The results indicated that high levels of xylanase can be produced in rice plants.

  16. 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.

  17. [Xylanase and cellulase of fungus Cerrena unicolor VKM F-3196: production, properties, and applications for the saccharification of plant material].

    Science.gov (United States)

    Belova, O V; Lisov, A V; Vinokurova, N G; Kostenevich, A A; Sapunova, L I; Lobanok, A G; Leont'evskiĭ, A A

    2014-01-01

    Under the conditions of submerged cultivation in a medium containing microcrystalline cellulose, the Cerrena unicolor VKM F-3196 basidiomycete is capable of producing xylanase and cellulase. Electrophoretically homogeneous cellulase and xylanase were obtained using ion exchange and hydrophobic chromatography. The molecular weight of both cellulase and xylanase was -44 kDa. It was shown that xylanase catalyzed the hydrolysis of xylan with the production of xylose, xylobiose, and xylotetrose and it exhibited properties of endoxylanases. Cellulase hydrolyzed carboxymethylcellulose, xylan, and microcrystalline cellulose with the formation of cellotriose and cellotetraose. For both enzymes, the pH optimum was -4.0. The enzymes exhibited moderate thermostability: xylanase retained 35% of the initial activity for an hour at 60 degrees C; cellulase, 10% under the same conditions. Xylanase, cellulose, and a mixture of these enzymes saccharified plant material (wheat, rye, wheat middling, and oat), indicating the possible use of these enzymes in biotechnology.

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

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    Ibrahim Che Omar

    2005-03-01

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

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

    Science.gov (United States)

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

    2013-08-01

    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) and from Penicillium funiculosum (PfXynC) were heterologously expressed in Pichia pastoris. The PoXyn3 and PfXynC cDNAs encoding mature xylanases were cloned into pGAPZαA vectors and integrated into the genome of P. pastoris X-33 under the control of the glyceraldehyde 3-phosphate dehydrogenase constitutive promoter. PfXynC was expressed as a His-tagged recombinant protein and purified from the supernatant homogeneity by a one-step purification protocol using immobilized metal affinity chromatography. The recombinant PoXyn3 was purified using a single anion-exchange chromatography. The purified recombinant enzymes were optimally active at 45°C and pH 4.0 for PoXyn3 and 40°C and pH 3.0 for PfXynC. The measured kinetic parameters (k(cat) and Vmax) showed that PfXynC was five times more active than PoXyn3 irrespective of the substrate whereas the apparent affinity (K(m)) was similar. The recombinant enzymes showed distinct sensitivity to the Triticum aestivum xylanase inhibitor TAXI-I.

  20. Efficiency of different xylanase preparations in diets for pekin ducks.

    Science.gov (United States)

    Timmler, R; Rodehutscord, M

    2001-01-01

    Four experiments were conducted with a total of 2288 pekin ducks. Day-old ducklings were group-penned on straw bedding and were fed complete, pelleted diets ad libitum for up to 49 days depending on experiment. In each experiment, starter diets (until day 21) and grower diets (from day 22) were used adequate in ME content and nutrient content. The sum of wheat, rye, and triticale amounted to at least 57% (starter diet) and 63% (grower diet), respectively. The inclusion level of wheat, rye, and triticale was different between experiments, with a maximum rye inclusion of 45%. Five different enzyme preparations all having, 1,4-beta-xylanase as the main activity were considered in this study with either one (2 preparations) or three (3 preparations) levels of supplementation. The effect of enzyme supplementation on ileal digesta viscosity was studied at the end of two experiments comprising 4 enzyme preparations. A significant reduction in digesta viscosity was determined for all preparations. The viscosity of digesta was higher in birds that were fed 45% rye in their diet as compared to those fed a diet based on triticale and wheat, even with enzyme supplementation. Differences in digesta viscosity were not reflected in growth or feed conversion data. In one experiment, the body weight of ducks on day 21 was significantly improved by enzyme supplementation. This effect disappeared with progress in experiment. In another experiment, feed intake was significantly improved with enzyme supplementation. Apart from this, no statistically significant improvement in performance could be detected. On overall average, the final BW of ducks fed an enzyme was (as compared to the unsupplemented control = 100), 100, and the feed conversion ratio was 101. There is no indication from the growth and feed conversion data that an enzyme effect becomes more pronounced with increasing inclusion rate of soluble NSP by rye. It is concluded that supplementary xylanases are efficient in

  1. 产木聚糖酶菌株的分离及木聚糖酶的固定化%Isolation of Xylanase Producing Strain and Immobilization of Xylanase

    Institute of Scientific and Technical Information of China (English)

    王瑞君

    2012-01-01

    从草鱼肠道中分离筛选出6株产木聚糖酶的菌株,从中挑选产酶量最大的菌株测定酶液活力,并采用壳聚糖、海藻酸钠等固定游离酶.结果表明,与游离酶相比,壳聚糖固定酶的最适pH向碱性方向偏移,海藻酸钠固定酶的最适pH向酸性方向偏移.与游离酶相比,固定化酶有较高的最适温度和较强的热稳定性.%6 xylanase producing strains were screened from the intestinal tract of grass carp, among -which the strain with the highest xylanase yield was adopted to detect the enzyme activity of xylanase. Furthermore, the xylanase was immobilized by chitosan and sodium alginate. The results showed that compared with free enzyme, the optimal pH for chitosan immobilized xylanase was higher while for sodium alginate immobilized xylanase was lower. The optimal temperature for the immobilized xylanase was higher than that of free enzyme. The immobilized xylanase also showed broader temperature adaptability.

  2. Stable production of thermotolerant xylanase B of Clostridium stercorarium in transgenic tobacco and rice.

    Science.gov (United States)

    Kimura, Tetsuya; Mizutani, Tomomi; Sun, Jia-Lin; Kawazu, Tetsu; Karita, Shuichi; Sakka, Makiko; Kobayashi, Yasuo; Ohmiya, Kunio; Sakka, Kazuo

    2010-01-01

    The xylanase B gene encoding a thermostable family 10 xylanase of Clostridium stercorarium was expressed in plants under the control of a constitutive promoter. Two forms of the xylanase B gene, the xynB gene encoding the full length of the xylanase B gene including the bacterial signal sequence and the xynBM gene without the signal sequence region, were introduced into tobacco BY-2 cells and tobacco plants respectively under the control of the cauliflower mosaic virus 35S promoter. Transgenic BY-2 cells and tobacco plants showed xylanase activity and normal growth. The recombinant enzyme produced in transgenic BY-2 cells harboring the xynB gene was secreted into the culture supernatant, and the recombinant enzyme produced in transgenic BY-2 cells harboring the xynBM gene was localized in the cells. In contrast to tobacco plants, expression of the xynB gene under the control of the rice actin promoter in rice plants was toxic to host cells. However, the recombinant XynBM accumulated in leaf cells, and no phenotypic effect of expression of the xynBM gene was observed. Enzyme activity was maintained in cell-free extracts of transgenic rice leaves at 60 degrees C for 72 h, and the recombinant XynBM degraded hemicellulosic polymers in cell-free extracts of transgenic rice leaves.

  3. Structure-Specificity Relationships of an Intracellular Xylanase from Geobacillus stearothermophilus

    Energy Technology Data Exchange (ETDEWEB)

    Solomon,V.; Teplitsky, A.; Shulami, S.; Zolotnitsky, G.; Shoham, Y.; Shoham, G.

    2007-01-01

    Geobacillus stearothermophilus T-6 is a thermophilic Gram-positive bacterium that produces two selective family 10 xylanases which both take part in the complete degradation and utilization of the xylan polymer. The two xylanases exhibit significantly different substrate specificities. While the extracellular xylanase (XT6; MW 43.8 kDa) hydrolyzes the long and branched native xylan polymer, the intracellular xylanase (IXT6; MW 38.6 kDa) preferentially hydrolyzes only short xylo-oligosaccharides. In this study, the detailed three-dimensional structure of IXT6 is reported, as determined by X-ray crystallography. It was initially solved by molecular replacement and then refined at 1.45 {angstrom} resolution to a final R factor of 15.0% and an R{sub free} of 19.0%. As expected, the structure forms the classical ({alpha}/{beta}){sub 8} fold, in which the two catalytic residues (Glu134 and Glu241) are located on the inner surface of the central cavity. The structure of IXT6 was compared with the highly homologous extracellular xylanase XT6, revealing a number of structural differences between the active sites of the two enzymes. In particular, structural differences derived from the unique subdomain in the carboxy-terminal region of XT6, which is completely absent in IXT6. These structural modifications may account for the significant differences in the substrate specificities of these otherwise very similar enzymes.

  4. Analysis of inducers of xylanase and cellulase activities production by Ganoderma applanatum LPB MR-56.

    Science.gov (United States)

    Salmon, Denise Naomi Xavier; Spier, Michele Rigon; Soccol, Carlos Ricardo; Vandenberghe, Luciana Porto de Souza; Weingartner Montibeller, Valesca; Bier, Mário César Jucoski; Faraco, Vincenza

    2014-08-01

    This manuscript describes the analysis of the effect of cellulose, carboxymethylcellulose (CMC), xylan, and xylose as inducers of cellulase and xylanase activity production by Ganoderma applanatum MR-56 and the optimization of their production in liquid cultures by statistical methods. The Plackett-Burman screening design was applied to identify the most significant inducers of xylanase and cellulase activities production by G. applanatum MR-56. The most significant effect on xylanase and cellulase activities production was exercised by cellulose, even if xylose and CMC were also effective at some times. The combined effect of cellulose, yeast extract, and pH was analyzed by a 2(3) factorial experimental design with four central points that showed that the maximum tested cellulose (1 % w/v) and yeast extract (5 g L(-1)) concentrations gave the maximum production of xylanase (8.24 U mL(-1)) and cellulase (3.29 U mL(-1)) activity at pH 6 and 4, respectively. These values achieved for cellulase and xylanase activity represent 12-25 fold and 36 fold higher values than the maximum so far reported for other strains of G. applanatum, respectively.

  5. 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.

  6. Isolation and properties of Aspergillus niger IBT-90 xylanase for bakery.

    Science.gov (United States)

    Romanowska, Irena; Polak, Jacek; Bielecki, Stanisław

    2006-02-01

    Xylanase of low molecular weight (K II) was isolated from the fungus Aspergillus niger IBT-90 cultivated in medium with wheat bran. K II was purified by precipitation with ammonium sulphate (20-80% saturation) and gel filtration on Biogel P-10. This enzyme is most active in hydrolysis of birchwood xylan at 50 degrees C and pH 5.5. Xylanase K II has an ability to degrade 1,4-beta-bonds and to debranch substrates. It degrades not only xylans but also cellulose, an important factor for its application in bakery. Ag+, Fe3+ and NBS are strong inhibitors of the enzyme. DTT and Na+ activate xylanase K II by 24 and 13%, respectively. Enzyme K II used as additive to flour improves dough properties, increases the volume of wheat-rye and whole meal bread, and increases the porosity of crumb and the moisture of the final product, consequently extending the shelf life of bread.

  7. 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.

  8. Characterization and high expression of recombinant Ustilago maydis xylanase in Pichia pastoris.

    Science.gov (United States)

    Han, Hongjuan; You, Shuang; Zhu, Bo; Fu, Xiaoyan; Sun, Baihui; Qiu, Jin; Yu, Chengye; Chen, Lei; Peng, Rihe; Yao, Quanhong

    2015-03-01

    A recombinant xylanase gene (rxynUMB) from Ustilago maydis 521 was expressed in Pichia pastoris, and the enzyme was purified and characterized. Phylogenetic analysis demonstrated that rxynUMB belongs to glycosyl hydrolase family 11. The Trp84, Trp95, Glu93, and Glu189 residues are proposed to be present at the active site. The apparent molecular mass of the recombinant xylananse was approximately 24 kDa, and the optimum pH and temperature were 4.3 and 50 °C, respectively. Xylanase activity was enhanced by 166 and 115% with Fe(2+) and Mn(2+), respectively. The biochemical properties of this recombinant xylanase suggest that it may be a useful candidate for a variety of commercial applications.

  9. Improvement of xylanase production by Penicillium canescens 10-10c in solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Thonart P.

    2008-01-01

    Full Text Available Among hemicellulases, xylanases are catalysts of considerable interest so as fundamental than applied point of view. However,it is paradoxical to note that the high cost of their production limits their use on a large scale. The use of purified xylan as culturesubstrate increases the production cost of the enzyme. Consequently, for commercial applications, it is advisable to developprocesses starting from inexpensive substrates. The purpose of this study is to optimise xylanases production in solid-statefermentation based on agricultural residues. The strain is Penicillium canescens 10-10c, selected in our laboratory for his abilityto produce xylanase activity free of cellulase. Assays concern optimization of different culture parameters in order to developin the future a solid-state fermentation reactor with soya oil cake. These parameters are: medium composition, temperatureincubation, induction and repression mechanisms. Soya oil cake in pellets (size > 10 mm gave a higher enzymatic activity.Great volume of culture medium reduced the enzymatic production. The presence of lactose, saccharose or starch of corn hasa positive effect on the production of xylanase while the presence of xylose, mannose, galactose, arabinose, cellobiose andpectin or methylcellulose reduces the production of xylanase. The sources of phosphorus (di-potassic and di-sodic enhancexylanase production. The enzymatic production obtained in Erlenmeyer flasks (250 ml after 7 days incubation at 30°C isabout 14 000 U.g-1 of carbon source. The nature of inoculum affects the enzymatic productivity. Indeed, better productivitywas obtained with inoculation by solid preculture (956 U.g-1 per day than liquid preculture (473 U.g-1 per day and sporessuspension (383 U.g-1 per day. These observed enzymatic activity levels are higher than those related in the literature, whichshows all the potentialities of this strain and this technique for the production of xylanase and allow to develop

  10. 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., & Suhartono, M. T. (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.

  11. 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 ; Sorgatto, Michele ; Nogueira, Simone de Peixoto; Betini, Jorge Henrique ; Zanoelo, Fabiana ; Marques, Maria ; Polizeli, Maria de Lourdes Teixeira de Moraes; 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 ...

  12. Stable expression of a thermostable xylanase of Clostridium thermocellum in cultured tobacco cells.

    Science.gov (United States)

    Kimura, Tetsuya; Mizutani, Tomomi; Sakka, Kazuo; Ohmiya, Kunio

    2003-01-01

    Two distinct domains of the xynA gene from Clostridium thermocellum encoding a xylanase catalytic domain (XynAl) and a xylanase catalytic domain with a cellulose binding domain (XynA2) under the control of the cauliflower mosaic virus 35S promoter were electroporated into cultured tobacco BY-2 cells. Transgenic BY -2 calli expressing xylan-hydrolyzing activity were obtained at high frequency for both genes. Western blot analysis using an anti-XynA antibody indicated that XynAl and XynA2 were produced in these calli.

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

    DEFF Research Database (Denmark)

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

    2003-01-01

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

  14. 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.

  15. STUDY ON CELLULASE FERMENT PRODUCTION OF CHAETOMIUM CUPREUM%角毛壳菌产纤维素酶条件的研究

    Institute of Scientific and Technical Information of China (English)

    张海燕; 杨谦

    2006-01-01

    对角毛壳菌(Chaetomium cupreum)产纤维素酶的发酵条件进行了研究.结果表明:Mendels培养基适于角毛壳菌产纤维素酶,CMC酶活可达到15.7 U,β-葡萄糖苷酶活达到了15.8 U,滤纸酶活达到12.1 U.最佳产酶碳源是微结晶纤维素.发酵156 h是角毛壳菌产滤纸酶活的最佳时间,达到了14.4 U,发酵180 h是角毛壳菌产CMC酶活和β-葡萄糖苷酶活的最佳时间,酶活可分别达到37.6 U和38.4 U.

  16. 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.

  17. Cloning, expression and applicability of thermo-alkali-stable xylanase of Geobacillus thermoleovorans in generating xylooligosaccharides from agro-residues.

    Science.gov (United States)

    Verma, Digvijay; Satyanarayana, T

    2012-03-01

    A xylanase gene (xyl-gt) of 1.224 kbp was cloned from the extremely thermophilic bacterium Geobacillus thermoleovorans that encodes a protein containing 408 amino acid residues. Eight conserved regions (signature sequences) of GH family 10 xylanases have been found in the xylanase. When the xylanase gene was cloned and expressed in Escherichia coli BL21 (DE3), the recombinant strain produced xylanase titer of 270 U mg(-1) which is 27-fold higher than the wild strain. It is optimally active at 80°C and pH 8.5 with a high thermostability over broad range of pH (6-12) and temperature (40-100°C). The end products of the hydrolysis of birch wood xylan and agro-residues included xylobiose, xylotriose, xylotetraose and xylopentaose. The xylanase of G. thermoleovorans is one of the rare xylanases that exhibits thermo-alkali-stability, and thus, it is a suitable candidate for pre-bleaching of paper pulps and generating xylooligosaccharides from agro-residues for use as prebiotics.

  18. Functional analysis of glycoside hydrolase family 8 xylanases shows narrow but distinct substrate specificities and biotechnological potential.

    Science.gov (United States)

    Pollet, Annick; Schoepe, Jan; Dornez, Emmie; Strelkov, Sergei V; Delcour, Jan A; Courtin, Christophe M

    2010-08-01

    The potential of glycoside hydrolase family (GH) 8 xylanases in biotechnological applications is virtually unexplored. Therefore, the substrate preference and hydrolysis product profiles of two GH8 xylanases were evaluated to investigate their activities and substrate specificities. A GH8 xylanase from an uncultured bacterium (rXyn8) shows endo action but very selectively releases xylotriose from its substrates. It has a higher activity than the Pseudoalteromonas haloplanktis GH8 endo-xylanase (PhXyl) on xylononaose and smaller xylo-oligosaccharides. PhXyl preferably degrades xylan substrates with a high degree of polymerization. It is sterically more hindered by arabinose substituents than rXyn8, producing larger end hydrolysis products. The specificities of rXyn8 and PhXyl differ completely from these of the previously described GH8 xylanases from Bifidobacterium adolescentis (BaRexA) and Bacillus halodurans (BhRex). As reducing-end xylose-releasing exo-oligoxylanases, they selectively release xylose from the reducing end of small xylo-oligosaccharides. The findings of this study show that GH8 xylanases have a narrow substrate specificity, but also one that strongly varies between family members and is distinct from that of GH10 and GH11 xylanases. Structural comparison of rXyn8, PhXyl, BaRexA, and BhRex showed that subtle amino acid changes in the glycon as well as the aglycon subsites probably form the basis of the observed differences between GH8 xylanases. GH8 xylanases, therefore, are an interesting group of enzymes, with potential towards engineering and applications.

  19. 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 ...

  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 the thermophilic mold Humicola lanuginosa in solid-state fermentation.

    Science.gov (United States)

    Kamra, Pankaj; Satyanarayana, T

    2004-11-01

    Among the lignocellulosic substrates tested, wheat bran supported a high xylanase (EC 3.2.1.8) secretion by Humicola lanuginosa in solid-state fermentation (SSF). Enzyme production reached a peak in 72 h followed by a decline thereafter. Enzyme production was very high (7832 U/g of dry moldy bran) when wheat bran was moistened with tap water at a substrate--to--moistening agent ratio of 1:2.5 (w/v) and an inoculum level of 3 x 106 spores/10 g of wheat bran at a water activity (aw) of 0.95. Cultivation of the mold in large enamel trays yielded a xylanase titer comparable with that in flasks. Parametric optimization resulted in a 31% increase in enzyme production in SSF. Xylanase production was approx 23-fold higher in SSF than in submerged fermentation (SmF). A threshold constitutive level of xylanase was secreted by H. lanuginosa in a medium containing glucose as the sole carbon source. The enzyme was induced by xylose and xylan. Enzyme synthesis was repressed beyond 1.0% (w/v) xylose in SmF, whereas it was unaffected up to 3.0% (w/w) in SSF, suggesting a minimization of catabolite repression in SSF.

  2. Improvement of the quality of wheat bread by addition of glycoside hydrolase family 10 xylanases.

    Science.gov (United States)

    Zheng, Han; Guo, Bing; Chen, Xiu-Lan; Fan, Sou-Jin; Zhang, Yu-Zhong

    2011-04-01

    Although many xylanases are widely used in the baking industry, only one glycoside hydrolase family 10 (GH 10) xylanase has previously been reported to be effective in baking. In this study, we compared the effectiveness of two GH 10 xylanases, psychrophilic XynA from Glaciecola mesophila and mesophilic EX1 from Trichoderma pseudokoningii, in bread making. The optimal dosages needed to improve wheat flour dough and bread quality were 270-U/kg flour for EX1 and 0.9-U/kg flour for XynA. At their optimal dosage, both XynA and EX1 had significant dough-softening ability, resulting in a 50% reduction in Brabender units. XynA was more effective than EX1 in reducing the time to reach maximum consistency. XynA and EX1 showed similar effects in improving the bread volume (~30% increase). EX1 was more effective in reducing the initial crumb firmness. Although both enzymes exhibited similar anti-staling effects on the bread, based on a decrease in the bread firmness, XynA had a greater effect on reducing the firming rate, and EX1 showed an enhanced reduction in the initial firmness. These results show that these two GH 10 xylanases have unique advantages in improving dough and bread quality and indicate their potential in bread making.

  3. 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 (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.

  4. [Biosynthesis of cellulolytic enzymes and xylanase during submerged cultivation of the fungus Aspergillus terreus 17P].

    Science.gov (United States)

    Loginova, L G; Guzhova, E P; Ismanlova, D Iu; Burdenko, L G

    1978-01-01

    The fungus Aspergillus terreus 17P--producer of cellulolytic enzymes--was cultivated in the Biotec 10 l fermenter on the medium containing minced and heated (at 200 degrees) wheat straw aerated with a different rate. At the mixing rate of 350 rpm and aeration rate of 0.7 r/rpm on the fourth day the culture liquid was obtained whose filtrate contained an active complex of cellulolytic enzymes and xylanase: CI--3.4; APB--1.1, Cx--35.7, cellobiase--0.23, xylanase--73.8 units/ml. The fractionation of the culture liquid filtrate with ammonium sulphate showed that the fraction precipitated at an interval of saturation of 0.3--0.7 contained the largest portion of cellulolytic enzymes and xylanase. The isolated enzymic preparations had a cellulolytic and xylanase activity and contained lipase, pectinase, laminarinase. They also contained low quantities of amylase, protease, beta-1,4- and beta-1,6-glucanase. Enzymic hylrolysis by the Asp. terreus 17P preparation of straw yielded glucose and xylose, of cotton, Na-KMC, cellobiose--glucose, Xylane hydrolyzate contained xylose and arabinose.

  5. Thermal Stability and Thermodynamics of Xylanase from Melanocarpus albomyces in Presence of Polyols and Salts

    Directory of Open Access Journals (Sweden)

    Gupteshwar Gupta

    2014-08-01

    Full Text Available An extracellular xylanase from the thermophilic fungus Melanocarpus albomyces IIS 68 was evaluated for its activity and stability in the presence of polyols and salts at 60 °C, and found to be an effective protecting agent for thermal deactivation of enzyme. Response surface methodology was employed to study the synergistic effects of glycerol and NaCl (thermo-stabilizers for xylanase stability. The addition of these thermo-stabilizers resulted in more than a 10-fold increment in enzyme half-life. Activation energy (Ea and thermodynamic parameters such as ∆H, ∆G, and ∆S were calculated for the thermal inactivation of free and immobilized xylanase. The immobilized enzyme underwent substantially less conformational changes because of its enhanced stability and increased compactness, providing better thermo-stability at elevated temperatures. These findings suggest that the combined effect of glycerol and sodium chloride serves as a potential stabilizer for extracellular thermophilic xylanase, which finds commercial application in many industries, especially in the pulp and paper industry.

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

    Digital Repository Service at National Institute of Oceanography (India)

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

    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...

  7. Characterization of a bifunctional xylanase/endoglucanase from yak rumen microorganisms.

    Science.gov (United States)

    Chang, Lei; Ding, Mozhu; Bao, Lei; Chen, Yingzhi; Zhou, Jungang; Lu, Hong

    2011-06-01

    A new gene, RuCelA, encoding a bifunctional xylanase/endoglucanase, was cloned from a metagenomic library of yak rumen microorganisms. RuCelA showed activity against xylan and carboxymethylcellulose (CMC), suggesting bifunctional xylanase/endoglucanase activity. The optimal conditions for xylanase and endoglucanase activities were 65°C, pH 7.0 and 50°C, pH 5.0, respectively. In addition, the presence of Co(+) and Co(2+) can greatly improve RuCelA's endoglucanase activity, while inhibits its xylanase activity. Further examination of substrate preference showed a higher activity against barley glucan and lichenin than against xylan and CMC. Using xylan and barley glucan as substrates, RuCelA displayed obvious synergistic effects with β-1,4-xylosidase and β-1,4-glucosidase. Generation of soluble oligosaccharides from lignocellulose is the key step in bioethanol production, and it is greatly notable that RuCelA can produce xylo-oligosaccharides and cello-oligosaccharides in the continuous saccharification of pretreated rice straw, which can be further degraded into fermentable sugars. Therefore, the bifunctional RuCelA distinguishes itself as an ideal candidate for industrial applications.

  8. Toevoeging van ß-glucanase en xylanase aan mengvoeders voor gespeende biggen

    NARCIS (Netherlands)

    Scholten, R.H.J.; Binnendijk, G.P.

    1997-01-01

    Op het proefbedrijf van het Proefstation voor de Varkenshouderij te Rosmalen is van november 1995 tot en met februari 1996 een onderzoek uitgevoerd om na te gaan wat de mogelijkheden zijn van toevoeging van de enzymen B-glucanase en xylanase (PorzymeSlOO@) aan biggenrantsoenen

  9. Marine-derived fungus Aspergillus cf. tubingensis LAMAI 31: a new genetic resource for xylanase production.

    Science.gov (United States)

    Dos Santos, Juliana A; Vieira, Juliana M F; Videira, Alexandre; Meirelles, Lucas A; Rodrigues, André; Taniwaki, Marta H; Sette, Lara D

    2016-03-01

    Marine-derived fungi have been reported as relevant producers of enzymes, which can have different properties in comparison with their terrestrial counterparts. The aim of the present study was to select from a collection of 493 marine-derived fungi the best producer of xylanase in order to evaluate the enzymatic production under different conditions. A total of 112 isolates produced xylanase in solid medium containing xylan as the carbon source, with 31 of them able to produce at least 10 U/mL of the enzyme. The best production (49.41 U/mL) was achieved by the strain LAMAI 31, identified as Aspergillus cf. tubingensis. After confirming the lack of pathogenicity (absence of ochratoxin A and fumonisin B2 production) this fungus was submitted to the experimental design in order to evaluate the effect of different variables on the enzymatic production, with the aim of optimizing culture conditions. Three experimental designs (two Plackett-Burman and one factorial fractional) were applied. The best condition for the enzymatic production was defined, resulting in an increase of 12.7 times in comparison with the initial production during the screening experiments. In the validation assay, the peak of xylanase production (561.59 U/mL) was obtained after 96 h of incubation, being the best specific activity achieved after 72 h of incubation. Xylanase from A. cf. tubingensis LAMAI 31 had optimum pH and temperature at 5.0 and 55 °C, respectively, and was shown to be stable at a range of 40-50 °C, and in pH from 3.6 to 7.0. Results from the present work indicate that A. cf. tubingensis LAMAI 31 can be considered as a new genetic resource for xylanase production.

  10. 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.

  11. 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.

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

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    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.

  13. A comparison of plate assay methods for detecting extracellular cellulase and xylanase activity.

    Science.gov (United States)

    Meddeb-Mouelhi, Fatma; Moisan, Jessica Kelly; Beauregard, Marc

    2014-11-01

    Identification of microorganisms for the production of carbohydrolytic enzymes is extremely important given the increased demand for these enzymes in many industries. To this end, dye-polysaccharide interactions which provide a visual indication of polymer hydrolysis (clear zones or halos) have been used for decades. For the detection of extracellular cellulase or xylanase activity many laboratories use Gram's iodine as the chromogenic dye, as it is a more rapid initial screening method compared to the use of other dyes. Here, we compared Gram's iodine and Congo red as indicators of polysaccharide hydrolysis. We attempted to detect cellulase activity using carboxymethylcellulose, and xylanase activity using birchwood xylan, in fourteen uncharacterized bacteria isolated from wood chips. Our results indicate that Gram's iodine may lead to identification of false positives in a typical screening protocol and that Congo red allows for avoidance of such pitfall. Congo red allowed detection of cellulase activity from live microbial colonies but not Gram's iodine. To confirm this, detection of enzymatic activity was also assessed using cell-free enzyme preparations. Congo red was found to be reliable in detecting cellulase activity with isolated enzymes preparations. Under the same conditions, neither of these dyes detected xylanase activity, despite independent evidence of xylanase activity for one of the preparations. We detected xylanase activity for this particular enzyme preparation using a coloured derivative of xylan (Remazol Brillant Blue R-xylan adduct) that respond to xylan hydrolysis. Our results suggest that methods that rely on interactions between a dye (Congo red or Gram's iodine) and a polymeric substrate (carboxymethylcellulose or birchwood xylan) for indirect detection of hydrolysis may require the use of relevant controls and independent confirmation of enzymatic activities.

  14. Optimization of Production Xylanase from Marine Bacterium Bacillus safensis P20 on Sugarcane Baggase by Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Nanik Rahmani

    2014-01-01

    Full Text Available Endo-1, 4-β-xylanase commonly called xylanase is an industrially important enzyme which degrades of lignocellulosic materials to sugar, alcohol and other useful product. The use of commercially xylan is too expensive for use at industrial scale production. For commercial applications, xylanases should ideally be produced from simple and inexpensive substrates. Indonesia has abundantly agro-residues such as sugar cane bagasse which is attractive to be used as carbon sources for the production of enzyme.  In this study, optimization of fermentation condition extracellular xylanasefrom marine bacterium, Bacillus safensisP20 has been conducted by using sugarcane bagasse as carbon source under sub merged fermentation (SMF. Maximum xylanase production was obtained at sugar cane bagasse concentration 1.5%, pH medium 7, and temperature fermentation 20oC, lactose as a carbone source and urea as a nitrogen source with activity 4.06 U/mL for 96 hours.

  15. 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.

  16. Research on microorganism xylanases and their applications%微生物木聚糖酶及其应用

    Institute of Scientific and Technical Information of China (English)

    高艳秀; 陈复生; 丁长河

    2012-01-01

    Xylan is the major constituent of hemicelluloses. Due to the structural heterogeneity of the xylans, xylan-degrading enzyme systems include several hydrolytic enzymes. Xylanase(EC 3.2.1.8,1,4-β-D-xylanase)can hydrolyze β-1,4-glycosidic linkages of the xylan backbone to produce short chain xylooligosacchrides of various lengths. Hence, endo-β - xylanase is the crucial enzyme component of microbial xylanolytic enzyme systems. And the microorganism xylanases system, classification, source and distribution were summarized. The microorganism xylanases including their properties, production and their applications in food, papermaking, feed industries were reviewed.%木聚糖(Xylan)是植物半纤维素的主要成分,是一种复杂的多聚五碳糖.木聚糖酶(Xylanase,EC 3.2.1.8)以内切方式作用于木聚糖主链,产生不同链长的寡糖和少量的木糖,是木聚糖降解酶系中最为关键的酶.本文综述了微生物木聚糖酶系统、微生物木聚糖酶的分类及其来源分布、微生物木聚糖酶的特性、产生及在食品、造纸、饲料行业的应用.

  17. Investigating the expression of F10 and G11 xylanases in Aspergillus niger A09 with qPCR.

    Science.gov (United States)

    Cui, Shixiu; Wang, Tianwen; Hu, Hong; Liu, Liangwei; Song, Andong; Chen, Hongge

    2016-09-01

    There exist significant differences between the 2 main types of xylanases, family F10 and G11. A clear understanding of the expression pattern of microbial F10 and G11 under different culture conditions would facilitate better production and industrial application of xylanase. In this study, the fungal xylanase producer Aspergillus niger A09 was systematically investigated in terms of induced expression of xylanase F10 and G11. Results showed that carbon and nitrogen sources could influence xylanase F10 and G11 transcript abundance, with G11 more susceptible to changes in culture media composition. The most favorable carbon and nitrogen sources for high G11 and low F10 production by A. niger A09 were xylan (2%) and (NH4)2C2O4 (0.3%), respectively. Following cultivation at 33 °C for 60 h, the highest xylanase activity (1132 IU per gram of wet mycelia) was observed. On the basis of differential gene expression of F10 and G11, as well as their different properties, we deduced that the F10 protein initially targeted xylan and hydrolyzed it into fragments including xylose, after which xylose acted as the inducer of F10 and G11 gene expression. These speculations also accounted for our failure to identify conditions favoring the high production of F10 but a low production of G11.

  18. Mutation breeding of antibiotics-production strain Chaetomium cupreum CH21%产抗生素角毛壳菌CH21的诱变育种

    Institute of Scientific and Technical Information of China (English)

    王俊; 谭红; 钟娟; 周金燕; 舒丹; 杨杰

    2011-01-01

    In order to obtain mutants with higher antibiotics yield, the germinating spores of the wild strain Chaetomium cupreum CH21 were mutated by two mutagens, microwave and UV, and screened by PDA double-plate and bioassay both using Colletotrichum capsici as indicator. The optimal mutagen dosages were determined: 500W 2450MHz under microwave irradiation for 15s, and 15W, 30cm under ultraviolet ray irradiation for 300s. CH21, the diameter of inhibiting ring of which was 12.50mm, was mutated by microwave and UV each for once. Five mutants with high yield of antibiotics were isolated. The diameter of inhibiting ring of CH21-2-20 was 20.20 mm, which was the largest and increased by 62% in comparison to CH21. Cultivated for five continuous generations, the diameter of inhibiting ring of CH21-2-20 stabilized between 19.40 mm and 20.38 mm.%以角毛壳菌(Chaetomium cupreum)CH21的萌发孢子作为诱变材料,采用微波辐照和紫外线照射进行诱变处理,用辣椒炭疽菌(Colletotrichum capsici)作为测试菌株,结合PDA双层平板法初筛和管碟法复筛,选育高产抗生素的突变株.结果表明:最佳微波诱变条件为500W、2450MHz、15s,紫外诱变条件为15W、30cm、300s.对出发菌株CH21进行一轮微波诱变和一轮紫外诱变,筛选得到5株高产菌株,其中CH21-2-20抑菌圈直径最大,为20.20mm,较抑菌圈直径为12.50mm的出发菌株增大了62%.连续传代5次,CH21-2-20抑菌圈直径稳定在19.40~20.38mm.

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

    Institute of Scientific and Technical Information of China (English)

    Ranganathan KAPILAN; Vasanthy ARASARATNAM

    2011-01-01

    To optimize culture conditions for xylanase production by solid state fermentation (SSF) using Bacillus pumilus,with paddy husk as support,solid medium contained 200 g of paddy husk with 800 mL of liquid fermentation medium [xylan,20.0 g/L; peptone,2.0 g/L; yeast extract,2.5 g/L; K2HPO4,2.5 g/L; KH2PO4,1.0 g/L; NaCl,0.1 g/L; (NH4)2SO4,2.0 g/L,CaCl22H2O,0.005 g/L; MgCl2·6H2O,0.005 g/L; and FeCl3,0.005 g/L] at pH 9.0 was applied.The highest xylanase activity (142.0 +0.47 U/g DM] was obtained on the 6th day at 30℃.The optimized paddy husk to liquid fermentation medium ratio was 2∶9,and the optimized culture temperature was 40℃.When commercial Birchwood xylan was replaced with different concentrations of corncob,xylanase production was maximized (224.2 U/g DM) in the medium with 150 g/L corncob.Xylanase production was increased by sucrose,fructose and arabinose,whereas reduced by glucose,galactose,lactose and amylose.When organic nitrogen sources were replaced with locally available nitrogen sources such as groundnut powder or sesame seedcake powder or coconut seedcake powder or soy meal powder,the highest xylanase production (290.7 U/g DM) was obtained in the medium with soy meal powder and 16.0 g/L of soy meal powder was the optimum (326.5±0.34 U/g DM).Based on the optimization studies,B.pumilus produced 2.3 times higher xylanase activity.The medium cost was reduced from 2 458.3 to 178.3 SLR/kg and the total activity which could be obtained from 1 kg of the medium was increased from 48 624 to 220 253 Units.

  20. Production, purification and characterisation of alkali stable xylanase from Cellulosimicrobium sp. MTCC 10645

    Institute of Scientific and Technical Information of China (English)

    Rajashri D Kamble; Anandrao R Jadhav

    2012-01-01

    Objective: The aim of this experimental study was production, purification and characterization of alkali stable xylanase from locally isolated Cellulosimicrobium sp. MTCC 10645, which is an important industrial enzyme used in the pulp and paper industry. Methods: The enzyme was produced in Erlenmeyer flasks containing fresh basal salt medium supplemented with 1% oat spelt xylan. The enzyme was extracted and isolated using ammonium sulphate precipitation and dialysis. It was further purified using DEAE cellulose chromatography and purity was checked by SDS-PAGE. Effect of temperature and pH on activity and stability of enzyme was studied. The enzyme was laso studied for its substrate specificity and kinetic parameters. Results: The isolate was identified on the basis of cultural, morphological, physiological and biochemical properties as well as 16S rRNA sequencing. Among the carbon sources tested, birchwood xylan found prominent for increased level of xylanase i. e. 96.33 U/ml. The enzyme was purified by DEAE cellulose chromatography at NaCl concentration of 0.25 M and had a molecular mass of 78.0 kDa. Xylanase was purified sixteen fold with a specific activity of 246.6 U/mg. Xylanase activity was maximum at 50℃. The enzyme was thermostable retaining 8%of the original activity after incubation at 60℃ of 4 h. The enzyme was active over a pH range of 6.0-11.0, although its activity was optimal at pH 7.0. About 48.52% of the enzyme activity was retained after 4 h at pH 11.0. The enzyme was active on oat spelt and birchwood xylans but not on avicel, CMC, cellobiose, starch or p-nitrophenyl xylopyranoside. The xylanase had Km and Vmax values of 4.76 mg/ml and 232.5 μmol/min/mg, respectively when birchwood xylan used as substrate. Conclusions:The xylanase showed a unique pattern of xylan hydrolysis releasing a large amount of intermediate products (xylotriose and xylobiose) with small quantity of xylose. Some of these characteristics make this enzyme potentially

  1. Cloning of xylanase gene of Streptomyces flavogriseus in Escherichia coli and bacteriophage lambda-induced lysis for the release of cloned enzyme.

    Science.gov (United States)

    Srivastava, R; Ali, S S; Srivastava, B S

    1991-03-01

    The xylanase gene of Streptomyces flavogriseus was cloned in pUC8 plasmid and expressed in Escherichia coli lysogenic for lambda cI857. lambda-Induced lysis of E. coli at 42 degrees C allowed efficient release of cloned enzyme activity in extracellular environment. The xylanase gene was located in the 0.8-kb HindIII fragment and coded for 18,000 Mr xylanase.

  2. 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

  3. 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.

  4. 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.

  5. Toxicological studies on Thermomyces lanuginosus xylanase expressed by Fusarium venenatum, intended for use in food.

    Science.gov (United States)

    Pedersen, P B; Broadmeadow, A

    2000-09-01

    The xylanase used in this study was produced by a submerged fermentation of Fusarium venenatum and contained a gene code originating from Thermomyces lanuginosus. The enzyme was subject to a 13-week toxicological test in rats and in vitro tests to document its safety in use. The enzyme is to be applied as a processing aid in the baking industry to improve handling and stability of dough. The enzyme was not found to be mutagenic in the Salmonella typhimurium reverse mutation assay, nor did it cause chromosomal aberrations in cultured human lymphocytes. Oral administration to rats of up to 10.0 ml/kg bw/day (equivalent to a Total Organic Solids dosage of 1.12 g/kg bw/day or a xylanase dosage of 89422 FXU (W)/kg bw/day) for 13 weeks did not cause any adverse effect.

  6. 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.

  7. 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

  8. 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%).

  9. Effects of disruption of xylanase-encoding genes on the xylanolytic system of Streptomyces lividans.

    Science.gov (United States)

    Arhin, F F; Shareck, F; Kluepfel, D; Morosoli, R

    1994-01-01

    Wild-type Streptomyces lividans produced the three xylanases (XlnA, XlnB, and XlnC) when xylan, xylan hydrolysates obtained by the action of XlnA, XlnB, and XlnC, or purified small xylo-oligosaccharides (xylobiose [X2], xylotriose [X3], xylotetraose [X4], and xylopentaose [X5]) were used as the carbon source. The three xylanase genes of S. lividans (xlnA, xlnB, and xlnC) were disrupted by using vectors that integrate into the respective genes. Disruption of one or more of the xln genes resulted in reduced growth rates and reduced total xylanase activities when the strain was grown in xylan. The greatest effect was observed when xlnA was disrupted. In medium containing xylan, disruption of xlnA did not affect expression of xlnB and xlnC; disruption of xlnB did not affect expression of xlnA but affected expression of xlnC; and disruption of xlnC did not affect expression of xlnA but affected expression of xlnB. A fraction of XlnB or XlnC hydrolytic products (those with a degree of polymerization greater than 11 [X11]) was found to stimulate expression of xlnB and xlnC in strains disrupted in xlnC and xlnB, respectively, whereas lower-molecular-weight fractions as well as purified small xylo-oligosaccharides did not. The stimulating molecule(s) lost its effect when it was hydrolyzed further by XlnA. A mechanism of transglycosylation reactions by the S. lividans xylanases is postulated to be involved in the regulation of xln genes. Images PMID:8051006

  10. Cloning, overexpression, and characterization of a novel alkali-thermostable xylanase from Geobacillus sp. WBI.

    Science.gov (United States)

    Mitra, Suranjita; Mukhopadhyay, Bidhan Chandra; Mandal, Anisur Rahaman; Arukha, Ananta Prasad; Chakrabarty, Kuheli; Das, Gourab Kanti; Chakrabartty, Pran Krishna; Biswas, Swadesh Ranjan

    2015-04-01

    An endo-β-1,4-xylanase gene xynA of a thermophilic Geobacillus sp. WBI from "hot" compost was isolated by PCR amplification. The gene encoding 407 residues were overexpressed in E. coli and purified by Ni-NTA chromatography. The purified enzyme (47 kDa) had a broad pH optimum of 6.0 to 9.0, and was active between 50 and 90 °C. The enzyme retained 100% of its activity when incubated at 65 °C for 1 h under alkaline condition (pH 10.0) and retained 75% activity at pH 11.0. The K(m) and V(max) of the enzyme were 0.9 mg ml(-1) and 0.8 µmol ml(-1) min(-1), respectively. In molecular dynamics simulation at 338 K (65 °C), the enzyme was found to be stable. At an elevated temperature (450 K) specific α-helix and β-turns of the proteins were most denatured. The denaturation was less in WBI compared with its highest homolog G. stearothermophilus T-6 xylanase with difference of six residues. The results predict that these regions are responsible for the improved thermostability observed over related enzymes. The present work encourages further experimental demonstration to understand how these regions contribute thermostability to WBI xylanase. The study noted that WBI produces a xylanase with unique characteristics, specifically alkali-thermostability.

  11. TLXI, a novel type of xylanase inhibitor from wheat (Triticum aestivum) belonging to the thaumatin family.

    Science.gov (United States)

    Fierens, Ellen; Rombouts, Sigrid; Gebruers, Kurt; Goesaert, Hans; Brijs, Kristof; Beaugrand, Johnny; Volckaert, Guido; Van Campenhout, Steven; Proost, Paul; Courtin, Christophe M; Delcour, Jan A

    2007-05-01

    Wheat (Triticum aestivum) contains a previously unknown type of xylanase (EC 3.2.1.8) inhibitor, which is described in the present paper for the first time. Based on its >60% similarity to TLPs (thaumatin-like proteins) and the fact that it contains the Prosite PS00316 thaumatin family signature, it is referred to as TLXI (thaumatin-like xylanase inhibitor). TLXI is a basic (pI> or =9.3 in isoelectric focusing) protein with a molecular mass of approx. 18-kDa (determined by SDS/PAGE) and it occurs in wheat with varying extents of glycosylation. The TLXI gene sequence encodes a 26-amino-acid signal sequence followed by a 151-amino-acid mature protein with a calculated molecular mass of 15.6-kDa and pI of 8.38. The mature TLXI protein was expressed successfully in Pichia pastoris, resulting in a 21-kDa (determined by SDS/PAGE) recombinant protein (rTLXI). Polyclonal antibodies raised against TLXI purified from wheat react with epitopes of rTLXI as well as with those of thaumatin, demonstrating high structural similarity between these three proteins. TLXI has a unique inhibition specificity. It is a non-competitive inhibitor of a number of glycoside hydrolase family 11 xylanases, but it is inactive towards glycoside hydrolase family 10 xylanases. Progress curves show that TLXI is a slow tight-binding inhibitor, with a K(i) of approx. 60-nM. Except for zeamatin, an alpha-amylase/trypsin inhibitor from maize (Zea mays), no other enzyme inhibitor is currently known among the TLPs. TLXI thus represents a novel type of inhibitor within this group of proteins.

  12. Computational design of an endo-1,4-[beta]-xylanase ligand binding site

    Energy Technology Data Exchange (ETDEWEB)

    Morin, Andrew; Kaufmann, Kristian W.; Fortenberry, Carie; Harp, Joel M.; Mizoue, Laura S.; Meiler, Jens (Vanderbilt)

    2012-09-05

    The field of computational protein design has experienced important recent success. However, the de novo computational design of high-affinity protein-ligand interfaces is still largely an open challenge. Using the Rosetta program, we attempted the in silico design of a high-affinity protein interface to a small peptide ligand. We chose the thermophilic endo-1,4-{beta}-xylanase from Nonomuraea flexuosa as the protein scaffold on which to perform our designs. Over the course of the study, 12 proteins derived from this scaffold were produced and assayed for binding to the target ligand. Unfortunately, none of the designed proteins displayed evidence of high-affinity binding. Structural characterization of four designed proteins revealed that although the predicted structure of the protein model was highly accurate, this structural accuracy did not translate into accurate prediction of binding affinity. Crystallographic analyses indicate that the lack of binding affinity is possibly due to unaccounted for protein dynamics in the 'thumb' region of our design scaffold intrinsic to the family 11 {beta}-xylanase fold. Further computational analysis revealed two specific, single amino acid substitutions responsible for an observed change in backbone conformation, and decreased dynamic stability of the catalytic cleft. These findings offer new insight into the dynamic and structural determinants of the {beta}-xylanase proteins.

  13. 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.

  14. Gene cloning and characterization of an acidic xylanase from Acidobacterium capsulatum.

    Science.gov (United States)

    Inagaki, K; Nakahira, K; Mukai, K; Tamura, T; Tanaka, H

    1998-06-01

    The gene xynA encoding an acid endo-beta-1,4-xylanase from an acidophilic bacterium, Acidobacterium capsulatum 161, was cloned and expressed in Eschrichia coli. The nucleotide sequence of the 1.6-kb DNA fragment containing xynA was analyzed, revealing an open reading frame of 1,215 bp encoding a peptide of 405 amino acid residues. The deduced amino acid sequence of XynA was very similar to other xylanases that are from the glycosyl hydrolase family 10. XynA was purified to homogeneity by SDS-polyacrylamide gel electrophoresis from E. coli transformants. The molecular mass and isoelectric point of XynA were 41 kDa and 7.3, respectively. The xylanase activity of the cloned XynA is an endo-acting enzyme that shows optimal activity at pH 5.0 and 65 degrees C, and is stable pH between 3.0 and 8.0. The K(m) and Vmax with oat spelt xylan as a substrate at pH 5.0 and 30 degrees C are 3.5 mg/ml and 403 mumol/min/mg.

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

    Directory of Open Access Journals (Sweden)

    Lončar Davor M.

    2016-01-01

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

  16. Screening of a xylanase clone from a fosmid library of rumen microbiota in Hu sheep.

    Science.gov (United States)

    Wang, Jiakun; Sun, Zhongyuan; Zhou, Yan; Wang, Qian; Ye, Jun'an; Chen, Zhenming; Liu, Jianxin

    2012-01-01

    The glycosyl hydrolase family 11, which is responsible for carbohydrate metabolism, was identified in the open reading frame (ORF) 6 of a xylanase positive clone from a fosmid library of rumen microbiota of Hu sheep. A BLASTP search of GenBank revealed that ORF6 encoded a 355-amino acid putative endoxylanase, having 61% similarity (e(-73)) to endo-1,4-β-xylanase of Fibrobacter succinogenes S85 (YP_003250510.1). Predicted with the SWISS-MODEL, there were two separate β-sandwich clusters linked with a high serine containing linker in ORF6. The N-terminal β-sandwich is a novel endoxylanase of the glycosyl hydrolase family 11 with a specific activity of 1150.00 U/mg. The optimal pH and temperature for this enzyme were shown to be pH 5.0 and 50°C, respectively. The C-terminal helped increase the stability of the xylanase but decreased the activity to some degree. The C-terminal β-sandwich could bind avicel, but no conserved domain could be found. It may be a novel carbohydrate-binding module.

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

    Science.gov (United States)

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

    2013-11-01

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

  18. 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

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

    Science.gov (United States)

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

    2014-10-01

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

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

    Directory of Open Access Journals (Sweden)

    Uma Gupta

    2009-12-01

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

  1. Cloning, expression and characteristics of a novel alkalistable and thermostable xylanase encoding gene (Mxyl retrieved from compost-soil metagenome.

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    Digvijay Verma

    Full Text Available 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 from DNA extracted from the compost-soil in the p18GFP vector, a clone (TSDV-MX1 was detected that exhibited clear zone of xylan hydrolysis on RBB xylan plate. The sequencing of 6.321 kb DNA insert and its BLAST analysis detected the presence of xylanase gene that comprised 1077 bp. The deduced protein sequence (358 amino acids displayed homology with glycosyl hydrolase (GH family 11 xylanases. The gene was subcloned into pET28a vector and expressed in E. coli BL21 (DE3. The recombinant xylanase (rMxyl exhibited activity over a broad range of pH and temperature with optima at pH 9.0 and 80°C. The recombinant xylanase is highly thermostable having T1/2 of 2 h at 80°C and 15 min at 90°C. CONCLUSION/SIGNIFICANCE: This is the first report on the retrieval of xylanase gene through metagenomic approach that encodes an enzyme with alkalistability and thermostability. The recombinant xylanase has a potential application in paper and pulp industry in pulp bleaching and generating xylooligosaccharides from the abundantly available agro-residues.

  2. 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

  3. Global and grain-specific accumulation of glycoside hydrolase family 10 xylanases in transgenic maize (Zea mays).

    Science.gov (United States)

    Gray, Benjamin N; Bougri, Oleg; Carlson, Alvar R; Meissner, Judy; Pan, Shihao; Parker, Matthew H; Zhang, Dongcheng; Samoylov, Vladimir; Ekborg, Nathan A; Michael Raab, R

    2011-12-01

    In planta expression of cell wall degrading enzymes is a promising approach for developing optimized biomass feedstocks that enable low-cost cellulosic biofuels production. Transgenic plants could serve as either an enzyme source for the hydrolysis of pretreated biomass or as the primary biomass feedstock in an autohydrolysis process. In this study, two xylanase genes, Bacillus sp. NG-27 bsx and Clostridium stercorarium xynB, were expressed in maize (Zea mays) under the control of two different promoters. Severe phenotypic effects were associated with xylanase accumulation in maize, including stunted plants and sterile grains. Global expression of these xylanases from the rice ubiquitin 3 promoter (rubi3) resulted in enzyme accumulation of approximately 0.01 mg enzyme per gram dry weight, or approximately 0.1% of total soluble protein (TSP). Grain-specific expression of these enzymes from the rice glutelin 4 promoter (GluB-4) resulted in higher-level accumulation of active enzyme, with BSX and XynB accumulating up to 4.0% TSP and 16.4% TSP, respectively, in shriveled grains from selected T0 plants. These results demonstrate the potential utility of the GluB-4 promoter for biotechnological applications. The phenotypic effects of xylanase expression in maize presented here demonstrate the difficulties of hemicellulase expression in an important crop for cellulosic biofuels production. Potential alternate approaches to achieve xylanase accumulation in planta without the accompanying negative phenotypes are discussed.

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

    Directory of Open Access Journals (Sweden)

    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.

  5. 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.

  6. Combination of Xylanase and Debranching Enzymes Specific to Wheat Arabinoxylan Improve the Growth Performance and Gut Health of Broilers.

    Science.gov (United States)

    Lei, Zhao; Shao, Yuxin; Yin, Xiaonan; Yin, Dafei; Guo, Yuming; Yuan, Jianmin

    2016-06-22

    Arabinoxylan (AX) is the major antinutritional factor of wheat. This study evaluated the synergistic effects of xylanase and debranching enzymes (arabinofuranosidase [ABF] and feruloyl esterase [FAE]) on AX. During in vitro tests, the addition of ABF or FAE accelerated the hydrolysis of water-soluble AX (WE-AX) and water-insoluble AX (WU-AX) and produced more xylan oligosaccharides (XOS) than xylanase alone. XOS obtained from WE-AX stimulated greater proliferation of Lactobacillus brevis and Bacillus subtilis than did fructo-oligosaccharides (FOS) and glucose. During in vivo trials, xylanase increased the average daily growth (ADG), decreased the feed-conversion ratio (FCR), and reduced the digesta viscosity of jejunum and intestinal lesions of broilers fed a wheat-based diet on day 36. ABF or FAE additions further improved these effects. Broilers fed a combination of xylanase, ABF, and FAE exhibited the best growth. In conclusion, the synergistic effects among xylanase, ABF, and FAE increased AX degradation, which improve the growth performance and gut health of broilers.

  7. Xylanases, Cellulases, and Acid Protease Produced by Stenocarpella maydis Grown in Solid-state and Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Edna María Hernández-Domínguez

    2014-03-01

    Full Text Available Activity levels of extracellular hydrolytic enzymes produced by Stenocarpella maydis, a fungal pathogen of maize, have so far not been reported. Production of xylanase, cellulase, and acid protease by this ascomycete using different culture media in solid-state and submerged fermentation was studied. In solid-state fermentation, polyurethane foam was used as an inert support, and corncob, corn leaves, and broken corn were used as biodegradable supports. The highest xylanase activity was produced in the medium with xylan in both fermentation systems, reaching 18,020 U/L and 19,266 U/L for submerged and solid-state fermentation, respectively. Cellulase production was observed only in the culture medium with carboxymethylcellulose, obtaining values of 7,872 U/L in submerged fermentation and 9,439 U/L in solid-state fermentation. The acid protease was produced only in minimal medium with glucose in acidic pH, reaching the highest levels of activity in SSF (806 U/L. The corncob was the best biodegradable support for the production of xylanases and acid protease. Two isoenzymes of xylanase and cellulase were observed in both fermentation systems, and three isoenzymes of xylanase were produced on the biodegradable supports.

  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. 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.

  10. Antioxidant activity of endophytic Chaetomium globosum from Eucommia ulmoides%杜仲内生球毛壳菌的抗氧化活性研究

    Institute of Scientific and Technical Information of China (English)

    谢辉; 陈双林

    2009-01-01

    本文研究了分离自杜仲叶片的内生真菌球毛壳菌菌株No.173发酵液提取物的抗氧化活性,采用铁氰化钾还原力测定法、β-胡萝卜素/亚油酸模型和光照核黄素体系评价了发酵液提取物的抗氧化作用,并采用Folin-Ciocalteu法测定杜仲内生球毛壳菌发酵液提取物总多酚含量.结果表明,其抗氧化能力与Vc基本相当;清除超氧阴离子的能力优于芦丁;多酚含量为255.53±1.38mg/g,是其主要的抗氧化活性成分.%Antioxidant activity of fermentation broth extracts of Chaetomium globosum strain No. 173 isolated from leaves of Eucommia ulmoides was studied. The reducing power determination, the bleaching of β-carotene suboleic acid model and the riboflavine photosensitization system were applied to evaluate the antioxidant effects of the extracts. Total phenolic content in the extracts was estimated by using the Folin-Ciocalteu colorimetric method. The results showed that the antioxidant capacity of the extracts was equivalent to Vc, and the capacity scavenging O2-. was better than rutin. The phenolic content of the extracts was 255.53±1.38mg/g (gallic acid equivalent).

  11. Studies on Mutation Breeding of High-Yielding Xylanase Strains by Low-Energy Ion Beam Implantation

    Institute of Scientific and Technical Information of China (English)

    LI Shichang; YAO Jianming; YU Zhengling

    2007-01-01

    As a new mutagenetic method,low-energy ion implantation has been used widely in many research areas in recent years.In order to obtain some industrial strains with high xylanase yield,the wild type strain Aspergillus niger A3 was mutated by means of nitrogen ions implantation (10 keV,2.6 × 1014~1.56 × 1015 ions/cm2) and a mutant N212 was isolated subsequently.However,it was found that the initial screening means of the high-yielding xylanase strains such as transparent halos was unfit for first screening.Compared with that of the wild type strain,xylanase production of the mutant N212 was increased from 320 IU/ml to 610 IU/ml,and the optimum fermentation temperature was increased from 28 ℃ to 30 ℃.

  12. 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

    ), water-unextractable arabinoxylan (WUAX), birchwood xylan, and wheat bran. Both the BsX and the BsX(mut) catalyzed the release of xylo-oligosaccharides with higher degree of polymerization from WUAX than from WEAX. At equimolar addition levels the activity of the BsX(mut) was lower than that of the Bs......X with respect to both the initial rate and the product yields obtained after prolonged reaction on the xylan substrates. The calculated substrate selectivity factors indicated that the BsX and the BsX(mut) both had higher catalytic rate on WUAX than on WEAX. Addition of a 100:1 (TAXI:xylanase) molar ratio...

  13. A Fusarium graminearum xylanase expressed during wheat infection is a necrotizing factor but is not essential for virulence.

    Science.gov (United States)

    Sella, Luca; Gazzetti, Katia; Faoro, Franco; Odorizzi, Silvana; D'Ovidio, Renato; Schäfer, Wilhelm; Favaron, Francesco

    2013-03-01

    Fusarium graminearum is the fungal pathogen mainly responsible for Fusarium head blight (FHB) of cereal crops, which attacks wheat spikes, reducing crop production and quality of grain by producing trichothecene mycotoxins. Several cytohistological studies showed that spike infection is associated with the production of cell wall degrading enzymes. Wheat tissue, as in other commelinoid monocot plants, is particularly rich in xylan which can be hydrolyzed by fungal endo-1,4-β-xylanase. The FG_03624 is one of the most expressed xylanase genes in wheat spikes 3 days after inoculation and was heterologously expressed in the yeast Pichia pastoris. The recombinant protein (22.7 kDa) possessed xylanase activity and induced cell death and hydrogen peroxide accumulation in wheat leaves infiltrated with 10 ng/μl or in wheat lemma surface treated with 20 ng/μl. This effect reflects that observed with other described fungal xylanases (from Trichoderma reesei, Trichoderma viride and Botrytis cinerea) with which the FG_03624 protein shares a stretch of amino acids reported as essential for elicitation of necrotic responses. Several F. graminearum mutants with the FG_03624 gene disrupted were obtained, and showed about 40% reduction of xylanase activity in comparison to the wild type when grown in culture with xylan as carbon source. However, they were fully virulent when assayed by single floret inoculation on wheat cvs. Bobwhite and Nandu. This is the first report of a xylanase able to induce hypersensitive-like symptoms on a monocot plant.

  14. Xylanase production by Penicillium canescens on soya oil cake in solid-state fermentation.

    Science.gov (United States)

    Antoine, Assamoi Allah; Jacqueline, Destain; Thonart, Philippe

    2010-01-01

    There is an increasing interest for the organic residues from various sectors of agriculture and industries over the past few decades. Their application in the field of fermentation technology has resulted in the production of bulk chemicals and value-added products such as amino acid, enzymes, mushroom, organic acids, single-cell protein, biologically active secondary metabolites, etc. (Ramachandran et al., Bioresource Technology 98:2000-2009, 2007). In this work, the production of extracellular xylanase by the fungus Penicillium canescens was investigated in solid-state fermentation using five agro-industrial substrates (soya oil cake, soya meal, wheat bran, whole wheat bran, and pulp beet). The best substrate was the soya oil cake. In order to optimize the production, the most effective cultivation conditions were investigated in Erlenmeyer flasks and in plastic bags with 5 and 100 g of soya oil cake, respectively. The initial moisture content, initial pH, and temperature of the culture affected the xylanase synthesis. The optimal fermentation medium was composed by soya oil cake crushed to 5 mm supplemented with 3% and 4% (w/w) of casein peptone and Na(2)HPO(4) x 2H(2)O. After 7 days of incubation at 30 degrees C and under 80% of initial moisture, a xylanase production level of 18,895 +/- 778 U/g (Erlenmeyer flasks) and 9,300 +/- 589 U/g (plastic bags) was reached. The partially purified enzyme recovered by ammonium sulfate fractionation was completely stable at freezing and refrigeration temperatures up to 6 months and reasonably stable at room temperature for more than 3 months.

  15. Biorefining of wood: combined production of ethanol and xylanase from waste fiber sludge.

    Science.gov (United States)

    Cavka, Adnan; Alriksson, Björn; Rose, Shaunita H; van Zyl, Willem H; Jönsson, Leif J

    2011-08-01

    The possibility to utilize fiber sludge, waste fibers from pulp mills and lignocellulose-based biorefineries, for combined production of liquid biofuel and biocatalysts was investigated. Without pretreatment, fiber sludge was hydrolyzed enzymatically to monosaccharides, mainly glucose and xylose. In the first of two sequential fermentation steps, the fiber sludge hydrolysate was fermented to cellulosic ethanol with the yeast Saccharomyces cerevisiae. Although the final ethanol yields were similar, the ethanol productivity after 9.5 h was 3.3 g/l/h for the fiber sludge hydrolysate compared with only 2.2 g/l/h for a reference fermentation with similar sugar content. In the second fermentation step, the spent fiber sludge hydrolysate (the stillage obtained after distillation) was used as growth medium for recombinant Aspergillus niger expressing the xylanase-encoding Trichoderma reesei (Hypocrea jecorina) xyn2 gene. The xylanase activity obtained with the spent fiber sludge hydrolysate (8,500 nkat/ml) was higher than that obtained in a standard medium with similar monosaccharide content (1,400 nkat/ml). Analyses based on deglycosylation with N-glycosidase F suggest that the main part of the recombinant xylanase was unglycosylated and had molecular mass of 20.7 kDa, while a minor part had N-linked glycosylation and molecular mass of 23.6 kDa. Chemical analyses of the growth medium showed that important carbon sources in the spent fiber sludge hydrolysate included xylose, small aliphatic acids, and oligosaccharides. The results show the potential of converting waste fiber sludge to liquid biofuel and enzymes as coproducts in lignocellulose-based biorefineries.

  16. 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.

  17. Cloning and characterization of the first GH10 and GH11 xylanases from Rhizopus oryzae.

    Science.gov (United States)

    Xiao, Zhizhuang; Grosse, Stephan; Bergeron, Hélène; Lau, Peter C K

    2014-10-01

    The only available genome sequence for Rhizopus oryzae strain 99-880 was annotated to not encode any β-1,4-endoxylanase encoding genes of the glycoside hydrolase (GH) family 10 or 11. Here, we report the identification and cloning of two such members in R. oryzae strain NRRL 29086. Strain 29086 was one of several selected fungi grown on wheat or triticale bran and screened for xylanase activity among other hydrolytic actions. Its high activity (138 U/ml) in the culture supernatant led to the identification of two activity-stained proteins, designated Xyn-1 and Xyn-2 of respective molecular masses 32,000 and 22,000. These proteins were purified to electrophoretic homogeneity and characterized. The specific activities of Xyn-1 and Xyn-2 towards birchwood xylan were 605 and 7,710 U/mg, respectively. Kinetic data showed that the lower molecular weight Xyn-2 had a higher affinity (K m=3.2 ± 0.2 g/l) towards birchwood xylan than Xyn-1 by about 4-fold. The melting temperature (T m) of the two proteins, estimated to be in the range of 49.5-53.7 °C indicated that they are rather thermostable proteins. N-terminal and internal peptide sequences were obtained by chemical digestion of the purified xylanases to facilitate cloning, expression in Escherichia coli, and sequencing of the respective gene. The cloned Rhizopus xylanases were used to demonstrate release of xylose from flax shives-derived hemicellulose as model feedstock. Overall, this study expands the catalytic toolbox of GH10 and 11 family proteins that have applications in various industrial and bioproducts settings.

  18. 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 cat......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...

  19. 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

    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......-d-xylanase and β-xylosidase activities; and XYL11 was a true xylanase characterized by high substrate specificity. These results indicate that F. commune with genetic modification is a promising source of enzymes for the decomposition of lignocellulosic biomass....

  20. Fungal cellulase/xylanase production and corresponding hydrolysis using pretreated corn stover as substrates.

    Science.gov (United States)

    Zhang, Liang; Wang, Xiaoqing; Ruan, Zhenhua; Liu, Ying; Niu, Xiaorui; Yue, Zhengbo; Li, Zhimin; Liao, Wei; Liu, Yan

    2014-01-01

    Three pretreated corn stover (ammonia fiber expansion, dilute acid, and dilute alkali) were used as carbon source to culture Trichoderma reesei Rut C-30 for cellulase and xylanase production. The results indicated that the cultures on ammonia fiber expansion and alkali pretreated corn stover had better enzyme production than the acid pretreated ones. The consequent enzymatic hydrolysis was performed applying fungal enzymes on pretreated corn stover samples. Tukey's statistical comparisons exhibited that there were significant differences on enzymatic hydrolysis among different combination of fungal enzymes and pretreated corn stover. The higher sugar yields were achieved by the enzymatic hydrolysis of dilute alkali pretreated corn stover.

  1. 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......-xylosidase activity was optimal at 65degreesC. SDS-PAGE and zymogram techniques revealed the presence of two xylan-degrading proteins of approximately 45 and 67 kDa in culture supernatants. To our knowledge, this paper is the first report on hemicellulose-degrading enzymes produced by an extremely halophilic archaeon....

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

    OpenAIRE

    Seyis,Isil; Aksoz, Nilufer

    2005-01-01

    The effect of some natural wastes (orange pomace, orange peel, lemon pomace, lemon peel, apple pomace, pear peel, banana peel, melon peel and hazelnut shell) on the production of xylanase from Trichoderma harzianum 1073 D3 has been studied and maximum activity has been observed on melon peel (26.5 U/mg of protein) followed by apple pomace and hazelnut shell. Also, molasses could be used as an additional carbon source as it decreased the production time approximately by 50 %. Finally, potentia...

  3. 嗜热毛壳菌(Chaetomium thermophile)液体发酵产生纤维素酶及酶学性质的研究%Studies on liquid-state fermentation for and properties of cellulase from Chaetomium thermophlie

    Institute of Scientific and Technical Information of China (English)

    路梅; 李多川

    2003-01-01

    探讨了嗜热真菌Chaetomium thermophile产生纤维素酶的液体发酵条件及滤纸酶(FPA)的特性.采用液体发酵培养法,通过对碳源、氮源、培养时间、培养基的起始pH值及产酶过程中pH值和蛋白质含量变化的研究发现:在2%纤维素、1%可溶性淀粉为碳源;2.0% KNO3+0.2%酵母粉为氮源;起始 pH值为6.5,50℃下培养9d后,各种酶活最高.发酵过程中,pH值和蛋白质的含量均在前3d下降,后升高.FPA的反应最适温度和pH值分别为60℃和5.5~6.0;且具有较高的热稳定性和pH稳定性.

  4. 嗜热多拟青霉发酵产物杀虫活性的初步研究%Preliminary Study on Insecticidal Activity of Zymotic Metabolites from Polypaecilum thermophilum D.M.Wang et D.C.Li

    Institute of Scientific and Technical Information of China (English)

    王冬梅

    2011-01-01

    以纯化的嗜热多拟青霉液体发酵培养,测定其代谢产物对棉蚜和温室白粉虱的毒杀效果.用水培棉苗法培养棉蚜,盆栽番茄培养温室白粉虱,采用菌株发酵产物浸渍法进行生物测定.结果表明:该代谢产物表现出较高的毒杀作用,室内生物测定72 h对棉蚜和温室白粉虱的校正死亡率分别为80.5%和66.3%.%The purifided Polypaecilum thermophilum D. M. Wang et D. C. Li was cultured by fermenta tion , and the effects of its zymotic metabolites on Aphis gossypii raised on water - planting cotton seedlings and Trialeurodes vaporariorum raised on potted tomato plants were studied. The bioassay was conducted through dipping leaves in fungi zymotic metabolites. The results showed that the zymotic metabolites had higher toxicity to Aphis gossypii and Trialeurodes vaporariorum with adjusted mortality of 80.5% and 66.3% respectively after bioassay for 72 hours.

  5. 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.

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

    Directory of Open Access Journals (Sweden)

    Ramanjaneyulu Golla

    2016-09-01

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

  7. Strategies of xylanase supplementation for an efficient saccharification and cofermentation process from pretreated wheat straw.

    Science.gov (United States)

    Alvira, Pablo; Tomás-Pejó, Elia; Negro, María José; Ballesteros, Mercedes

    2011-07-01

    Ethanol production from lignocellulosic raw materials includes a pretreatment step before enzymatic hydrolysis (EH). Pretreated substrates contain complex hemicelluloses in the solid fraction that can protect the cellulose from enzymatic attack. In addition, soluble xylooligomers are contained in the pretreated materials and may have an inhibitory effect on cellulase activity. In this context, several approaches for xylanase supplementation have been studied to increase EH yields. In this study, the whole slurry obtained after steam explosion pretreatment of wheat straw has been used as substrate. EH experiments were performed using commercial cellulase preparations supplemented with an endoxylanase (XlnC) from Aspergillus nidulans. Among different strategies of XlnC supplementation, the 24-h xylanase treatment before cellulase addition yielded an increase of 40.1 and 10.1% in glucose and xylose production, respectively. Different XlnC addition strategies were integrated in a simultaneous saccharification and cofermentation process (SSCF) using the xylose fermenting strain Saccharomyces cerevisiae F12. Ethanol production in SSCF was 28.4% higher when comparing to a simultaneous saccharification and fermentation process.

  8. 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.

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

    Directory of Open Access Journals (Sweden)

    BUDI SAKSONO

    2010-12-01

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

  10. 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 10 h 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.

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

    Directory of Open Access Journals (Sweden)

    L Sciessere

    2011-09-01

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

  12. 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.

  13. 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.

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

    Science.gov (United States)

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

    2013-12-01

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

  15. Crystallization and preliminary crystallographic analysis of endo-1,4-beta-xylanase I from Aspergillus niger

    NARCIS (Netherlands)

    Krengel, U; Rozeboom, HJ; Kalk, KH; Dijkstra, BW

    1996-01-01

    A family G xylanase from Aspergillus niger has been crystallized using the vapor-diffusion method. Several crystal forms could be obtained using various sodium salts as precipitants. Three of the crystal forms belong to space groups P2(1), P2(1)2(1)2(1) and P4(3) and have cell parameters of approxim

  16. 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.

  17. Fusion of carbohydrate binding modules from Thermotoga neapolitana with a family 10 xylanase from Bacillus halodurans S7.

    Science.gov (United States)

    Mamo, Gashaw; Hatti-Kaul, Rajni; Mattiasson, Bo

    2007-01-01

    Xylanase A of Thermotoga neapolitana contains binding domains both at the N- and C-terminal ends of the catalytic domain. In the N-terminal position it contains two carbohydrate-binding modules (CBM) which belong to family 22. These CBMs bind xylan but not to cellulose. The gene encoding the mature peptide of these CBMs was fused with an alkaline active GH10 xylanase from Bacillus halodurans S7 and expressed in Escherichia coli. The (His)(6) tagged hybrid protein was purified by immobilized metal affinity chromatography and characterized. Xylan binding by the chimeric protein was influenced by NaCl concentration and pH of the binding medium. Binding increased with increasing salt concentration up to 200 mM. Higher extent of binding was observed under acidic conditions. The fusion of the CBM structures enhanced the hydrolytic efficiency of the xylanase against insoluble xylan, but decreased the stability of the enzyme. The optimum temperature and pH for the activity of the xylanase did not change.

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

    Science.gov (United States)

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

  19. Seven N-terminal residues of a thermophilic xylanase are sufficient to confer hyperthermostability on its mesophilic counterpart.

    Directory of Open Access Journals (Sweden)

    Shan Zhang

    Full Text Available Xylanases, and especially thermostable xylanases, are increasingly of interest for the deconstruction of lignocellulosic biomass. In this paper, the termini of a pair of xylanases, mesophilic SoxB and thermophilic TfxA, were studied. Two regions in the N-terminus of TfxA were discovered to be potentially important for the thermostability. By focusing on Region 4, it was demonstrated that only two mutations, N32G and S33P cooperated to improve the thermostability of mesophilic SoxB. By introducing two potential regions into SoxB in combination, the most thermostable mutant, M2-N32G-S33P, was obtained. The M2-N32G-S33P had a melting temperature (Tm that was 25.6°C higher than the Tm of SoxB. Moreover, M2-N32G-S33P was even three-fold more stable than TfxA and had a Tm value that was 9°C higher than the Tm of TfxA. Thus, for the first time, the mesophilic SoxB "pupil" outperformed its thermophilic TfxA "master" and acquired hyperthermostability simply by introducing seven thermostabilizing residues from the extreme N-terminus of TfxA. This work suggested that mutations in the extreme N-terminus were sufficient for the mesophilic xylanase SoxB to acquire hyperthermostability.

  20. 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.

  1. Synthesis, processing and export of cytoplasmic endo-ß-1,4-xylanase from barley aleurone during germination

    NARCIS (Netherlands)

    Caspers, M.P.M.; Lok, F.; Sinjorgo, K.M.C.; Zeijl, M. van; Nielsen, K.A.; Cameron-Mills, V.

    2001-01-01

    We have identified the major endo-β-l,4-xylanase (XYN-1) in the aleurone of germinating barley grain, and show that it is expressed as a precursor of Mr 61 500 with both N- and C-terminal propeptides. XYN-1 is synthesized as an inactive enzyme in the cytoplasm, and only becomes active at a late stag

  2. 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.

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

    OpenAIRE

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

    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...

  4. Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris.

    Science.gov (United States)

    Sriyapai, Thayat; Somyoonsap, Peechapack; Matsui, Kenji; Kawai, Fusako; Chansiri, Kosum

    2011-05-01

    A thermophilic xylan-degrading Actinomadura sp. S14 was isolated from compost in Thailand. Hemicellulase activities such as endo-1,4-β-xylanase, β-xylosidase and α-arabinofuranosidase were induced with xylan-containing agriculture wastes and oat spelt xylan. The gene encoding xylanase consisting of 687bp was cloned from Actinomadura sp. S14. The deduced amino acid sequence contained a signal peptide of 41 amino acids and a probable mature xylanase of 188 amino acids. An open reading frame (xynS14) corresponding to a mature xylanase was expressed in Escherichia coli and Pichia pastoris. The specific activity of purified XynS14 (P. pastoris) was 2.4-fold higher than XynS14 (E. coli). Both XynS14s showed the same basic properties such as optimal pH and temperature (pH 6.0 and 80°C) and stability in a broad pH range (pH 5.0-11.0) and at high temperatures up to 80°C. Both XynS14s showed approximately the same substrate specificity and K(m) values toward various xylans, but XynS14 (P. pastoris) showed higher V(max) and K(cat) than XynS14 (E. coli). Higher specific activities of XynS14 (P. pastoris) may be due to protein-folding in the host. Purified XynS14 showed more endo-1,4-β-xylanase activity on xylan and xylooligosaccharides than on xylotriose.

  5. Xylan-binding xylanase Xyl30 from Streptomyces avermitilis: cloning, characterization, and overproduction in solid-state fermentation.

    Science.gov (United States)

    Hernández, Alberto; López, José C; Santamaría, Ramón; Díaz, Margarita; Fernández-Abalos, José M; Copa-Patiño, José L; Soliveri, Juan

    2008-06-01

    A DNA fragment from the lignocellulolytic actinomycete Streptomyces avermitilis CECT 3339 was cloned using a DNA probe from the xylanase gene xysA of Streptomyces halstedii. The nucleotide sequence analysis revealed two potential ORFs, xyl30 and hd30, encoding a deduced multimodular F/10 xylanase with a binding domain and a secreted glycoxyl hydrolase, respectively. In Streptomyces lividans carrying the subcloned DNA fragment, two xylanase activity bands with estimated molecular masses of 42.8 and 35 kDa (named Xyl30 forms "h" and "l", respectively), were detected by zymograms and SDS-PAGE. The two xylanases had identical N-terminal sequences, suggesting that Xyl30 "l" derived from Xyl30 "h" by C-terminal processing in the culture supernatant. No transcripts of hd30 were detected by RT-PCR. Characterization of the partially purified Xyl30 "h" confirmed the presence of a modular endoxylanase containing a xylan-binding domain, which after processing in the culture supernatant loses the aforementioned domain and thus its capacity to bind xylan (Xyl30 "l"). Xyl30 "h" achieved maximal activity at pH 7.5 and 60 degrees C, retaining more than 50% of its activity from pH 3 to 9 and more than 40% after a 1-h incubation at 70 masculineC. Moreover, in the recombinant host strain up to 400 U xylanase/g medium (dry weight) was produced in solid-state fermentation (SSF) using cereal bran as substrate. The high production yields of this enzyme and its biochemical features make it a good candidate for use in industrial applications.

  6. Energy and nutrient utilization of broiler chickens fed corn-soybean meal and corn-based diets supplemented with xylanase.

    Science.gov (United States)

    Stefanello, C; Vieira, S L; Carvalho, P S; Sorbara, J O B; Cowieson, A J

    2016-08-01

    A study was conducted to evaluate the effects of increased levels of a β-xylanase on energy and nutrient utilization of broiler chickens fed corn-soy diets. A total of 480 slow feathering Cobb × Cobb 500 male broilers were randomly distributed to 10 treatments having 8 replicates of 6 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 25 d. Two experimental diets, a conventional corn/soy-based basal diet (CS) and the basal diet in which 40% of the diet was displaced by corn (CN), were fed as-is or supplemented with 50, 100, 150, or 200 fungal β-xylanase units (FXU)/kg. Dietary treatments were distributed factorially as a 2 × 5 arrangement. Samples of feed, excreta, and ileal digesta were analyzed for determination of ileal digestible energy (IDE), metabolizable energy, and total tract retention of protein and lipid. No interactions between diet and xylanase were observed. The CS diets had higher (P energy utilization and nutrient digestibility when compared to the CN diets. AMEn and IDE were improved (P energy utilization and digestibility of crude protein and dry matter increased with xylanase supplementation in corn/soy-based diets. When xylanase was tested in the CS diet, 92 and 124 FXU/kg maximized the energy release effect; however, the maximum energy response in the CN diet or corn was not achieved until 200 FXU/kg.

  7. 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.

  8. Regulatory puzzle of xyn1 gene (xylanase1) expression in Trichoderma reesei

    Institute of Scientific and Technical Information of China (English)

    Robert L Mach; Elisabeth Würleitner; Astrid R Stricker; Roman Rauscher; Christian Wacenovsky

    2004-01-01

    @@ Xylanase 1 (Xyn1) is one of the two major representatives of the xylanase system of T. reesei; the mechanisms governing its expression were analysed throughout this study. All factors and regulatory motifs responsible for transcriptional regulation and the model of their interplay in induction and repression will be presented. Using in vivo foot printing analysis of xylan-induced and glucose repressed mycelia, we detected three adjacent nucleotide sequences contacted by DNA-binding proteins. Protection within the inverted repeat of the Cre1 (SYGGRG) consensus sequence on the non coding strand under repressing conditions is in perfect agreement with the previously reported Cre1dependent glucose repression of xyn1. Constitutive protein binding could be observed to a CCAAT-box and an inverted repeat of a 5′ GGCTAA 3′ sequence. EMSA with crude extracts from induced and repressed mycelia revealed that the latter motifs are sufficient for formation of the basal transcriptional complex under all conditions. The inverted repeat of GGCTAA closely resembles the consensus sequences of the cellulase and xylanase regulators Ace1, Ace2 and, Xyr1 (encoded by xyr1, cloned and characterised in this study) EMSA with heterologously expressed components of each factor and of the T. reesei Hap2/3/5 protein complex revealed that the basal transcriptional complex is formed by Xyr1and the Hap2/3/5. Additionally to the Cre1 mediated carbon catabolite repression a yet unknown mechanism antagonizing induction of xyn1 expression could be elucidated. Latter occurs through competition of the repressor Ace1 and Xyr1 for the GGCTAA motif. In vivo proof for the relevance of identified motifs could be given through analysis of T. reesei transformants containing correspondingly mutated versions of the xyn1 promoter fused to the A. niger goxA gene. The results indicated that the basal as well as the induction level of xyn1 gene transcription is dependent on an interaction of Xyr1with the

  9. An overview of the safety evaluation of the Thermomyces lanuginosus xylanase enzyme (SP 628) and the Aspergillus aculeatus xylanase enzyme (SP 578).

    Science.gov (United States)

    Bergman, A; Broadmeadow, A

    1997-01-01

    Xylanases SP 628 and SP 578 were produced by submerged fermentation of Aspergillus oryzae, containing a gene code originating from Thermomyces lanuginosus and Aspergillus aculeatus, respectively. Both enzymes were subject to the same series of toxicological tests to document their safety in use. The enzymes are to be applied as processing aids in the baking industry and in wheat starch separation. Neither enzyme was found to be mutagenic in the Salmonella typhimurium reverse mutation assay, nor did they cause chromosomal aberrations in cultured human peripheral lymphocytes. No evidence of inhalation toxicity or skin and eye irritation was found. The enzymes are not regarded as skin-sensitizers, although the Buehler test with guinea-pigs revealed a minor potential. Oral administration up to 10.0 ml/kg bw/day (equivalent to a Total Organic Solids amount of 13.3% for SP 628 and of 11.3% for SP 578) in 13-week rat studies did not show any adverse effect.

  10. Effect of xylanase and cellulase supplementation on growth performance, volatile fatty acids and caecal bacteria of broiler chickens fed with palm kernel meal-based diet

    OpenAIRE

    2014-01-01

    In this study, the effect of xylanase and cellulase supplementation in palm kernel meal (PKM) based diet on growth performance, volatile fatty acids (VFAs) and the caecal bacterial populations of broiler chickens were investigated. Seventy five day old male Cobb broiler chicks were randomly allocated to three dietary treatment groups receiving T1 (20% PKM-based diet without enzyme), T2 (20% PKM-based diet with xylanase) and T3 (20% PKM-based diet with cellulase). Each enzyme was supplemented ...

  11. Production of xylanase and protease by Penicillium janthinellum CRC 87M-115 from different agricultural wastes.

    Science.gov (United States)

    Oliveira, Luciana A; Porto, Ana L F; Tambourgi, Elias B

    2006-04-01

    Five agricultural wastes were evaluated in submerged fermentation for xylanolytic enzymes production by Penicillium janthinellum. The wastes were hydrolyzed in acid medium and the liquid fraction was used for cultivation. Corn cob (55.3 U/mL) and oat husk (54.8 U/mL) were the best inducers of xylanase. Sugar cane bagasse (23.0 U/mL) and corn husk (23.8 U/mL) were moderately good, while cassava peel was negligible. Protease production was very low in all agro-industrial residues. The maximum biomass yields were 1.30 and 1.17 g/L for cassava peel and corn husk after 180 h, respectively. Xylanolytic activity showed a cell growth associated profile.

  12. Alkalistable xylanase production by alkalitolerant Paenibacillus montaniterrae RMV1 isolated from red mud.

    Science.gov (United States)

    Arora, Amita; Krishna, Pankaj; Malik, Vinita; Reddy, M Sudhakara

    2014-10-01

    The alkalitolerant and xylanolytic bacterial strain (RMV1) isolated from red mud pond was identified as Paenibacillus montaniterrae based on both biochemical and 16S rDNA sequence analysis. The RMV1 bacterial isolate produced alkalistable and thermostable endoxylanase active over a broad range of pH (4.0-11.0) and temperature (20-100 °C), with optima at 50 °C and pH 9.0 with a T1/2 of 6.7 hours at 50 °C. This is the first report on the isolation of P. montaniterrae from bauxite residue with quite a high xylanase producing ability.

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

    Directory of Open Access Journals (Sweden)

    Isil Seyis

    2005-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Chun-Han Ko

    2013-02-01

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

  15. Cell Wall Degrading Enzymes Involved in Mycoparasitism of the Biocontrol Agent Chaetomium spirale ND35%生防因子螺旋毛壳ND35的细胞壁降解酶与重寄生作用

    Institute of Scientific and Technical Information of China (English)

    高克祥; 刘晓光; Dana Friesem; Leonid Chernin; 时呈奎

    2005-01-01

    Some Chaetomium spp. Are capable of antagonizing several plant pathogenic fungi through production of antibiotics and mycoparasitism. Secretion of lytic enzymes, mainly including glucanases and chitinases, is considered the most important step in the mycoparasitic process. In this study, an about 110kDa exo - β - 1,3 - glucanase from C. Spirale ND35 was detected both in culture filtrate and directly on PAGE and IEF gels, as well as chitinases, although protease was not detectable on Litmus milk agar plates. Coiling and penetrating the hyphae of host fungus Valsa mali were observed by scanning electron microscope (SEM), which may be related to the synergistic interaction between β - 1,3 - glucanase and chitinases. Β - 1,3 - glucanase activity of C. Spirale ND35 varied considerably when C. Spirale ND35 was grown in different carbon sources during various incubation time, and might be subjected to both induction by substrate and catabolite repression.

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

    Science.gov (United States)

    Todero Ritter, Carla Eliana; Camassola, Marli; Zampieri, Denise; Silveira, Mauricio Moura; Dillon, Aldo José Pinheiro

    2013-01-01

    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) (1.95  ±  0.04 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 (9.99 ± 0.75 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.

  17. Optimization of Xylanase Production through Response Surface Methodology by Fusarium sp. BVKT R2 Isolated from Forest Soil and Its Application in Saccharification

    Science.gov (United States)

    Ramanjaneyulu, Golla; Rajasekhar Reddy, Bontha

    2016-01-01

    Xylanses are hydrolytic enzymes with wide applications in several industries like biofuels, paper and pulp, deinking, food, and feed. The present study was aimed at hitting at high yield xylanase producing fungi from natural resources. Two highest xylanase producing fungal isolates—Q12 and L1 were picked from collection of 450 fungal cultures for the utilization of xylan. These fungal isolates—Q12 and L1 were identified basing on ITS gene sequencing analysis as Fusarium sp. BVKT R2 (KT119615) and Fusarium strain BRR R6 (KT119619), respectively with construction of phylogenetic trees. Fusarium sp. BVKT R2 was further optimized for maximum xylanase production and the interaction effects between variables on production of xylanase were studied through response surface methodology. The optimal conditions for maximal production of xylanase were sorbitol 1.5%, yeast extract 1.5%, pH of 5.0, Temperature of 32.5°C, and agitation of 175 rpm. Under optimal conditions, the yields of xylanase production by Fusarium sp. BVKT R2 was as high as 4560 U/ml in SmF. Incubation of different lignocellulosic biomasses with crude enzyme of Fusarium sp. BVKT R2 at 37°C for 72 h could achieve about 45% saccharification. The results suggest that Fusarium sp. BVKT R2 has potential applications in saccharification process of biomass. PMID:27713726

  18. 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.

  19. 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.

  20. 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.

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

    OpenAIRE

    L. dos Reis; Ritter,C. E. T.; R. C. Fontana; Camassola,M.; A. J. P. Dillon

    2015-01-01

    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 enzym...

  2. Properties of an alkali-thermo stable xylanase from Geobacillus thermodenitrificans A333 and applicability in xylooligosaccharides generation.

    Science.gov (United States)

    Marcolongo, Loredana; La Cara, Francesco; Morana, Alessandra; Di Salle, Anna; Del Monaco, Giovanni; Paixão, Susana M; Alves, Luis; Ionata, Elena

    2015-04-01

    An extracellular thermo-alkali-stable and cellulase-free xylanase from Geobacillus thermodenitrificans A333 was purified to homogeneity by ion exchange and size exclusion chromatography. Its molecular mass was 44 kDa as estimated in native and denaturing conditions by gel filtration and SDS-PAGE analysis, respectively. The xylanase (GtXyn) exhibited maximum activity at 70 °C and pH 7.5. It was stable over broad ranges of temperature and pH retaining 88 % of activity at 60 °C and up to 97 % in the pH range 7.5-10.0 after 24 h. Moreover, the enzyme was active up to 3.0 M sodium chloride concentration, exhibiting at that value 70 % residual activity after 1 h. The presence of other metal ions did not affect the activity with the sole exceptions of K(+) that showed a stimulating effect, and Fe(2+), Co(2+) and Hg(2+), which inhibited the enzyme. The xylanase was activated by non-ionic surfactants and was stable in organic solvents remaining fully active over 24 h of incubation in 40 % ethanol at 25 °C. Furthermore, the enzyme was resistant to most of the neutral and alkaline proteases tested. The enzyme was active only on xylan, showing no marked preference towards xylans from different origins. The hydrolysis of beechwood xylan and agriculture-based biomass materials yielded xylooligosaccharides with a polymerization degree ranging from 2 to 6 units and xylobiose and xylotriose as main products. These properties indicate G. thermodenitrificans A333 xylanase as a promising candidate for several biotechnological applications, such as xylooligosaccharides preparation.

  3. Identification of glutamic acid 78 as the active site nucleophile in Bacillus subtilis xylanase using electrospray tandem mass spectrometry.

    Science.gov (United States)

    Miao, S; Ziser, L; Aebersold, R; Withers, S G

    1994-06-14

    A new mechanism-based inactivator of beta-1,4-xylanases, 2',4'-dinitrophenyl 2-deoxy-2-fluoro-beta-xylobioside, has been synthesized and used to trap the covalent intermediate formed during catalysis by Bacillus subtilis xylanase. Electrospray mass spectrometry confirmed the 1:1 stoichiometry of the incorporation of inactivator into the enzyme. Inactivation of xylanase followed the expected pseudo-first-order kinetic behavior, and kinetic parameters were determined. The intermediate trapped was relatively stable toward hydrolytic turnover (t1/2 = 350 min). However, turnover could be facilitated by transglycosylation following the addition of the acceptor benzyl thio-beta-xylobioside, thus demonstrating the catalytic competence of the trapped intermediate. Reactivation kinetic parameters for this process of kre = 0.03 min-1 and Kre = 46 mM were determined. The nucleophilic amino acid was identified as Glu78 by a tandem mass spectrometric technique which does not require the use of radiolabels. The peptic digest of the labeled enzyme was separated by high-performance liquid chromatography and the eluent fed into a tandem mass spectrometer via an electrospray ionization device. The labeled peptide was identified as one of m/z = 826 (doubly charged) which fragmented in the collision chamber between the mass analyzers with loss of the mass of a 2-fluoroxylobiosyl unit. Confirmation of the peptide identity was obtained both by tandem mass spectrometric sequencing and by Edman degradation of the purified peptide. Glu78 is completely conserved in all members of this xylanase family and indeed is shown to be located in the active site in the recently determined X-ray crystal structure.

  4. 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.

  5. 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.

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

    Directory of Open Access Journals (Sweden)

    Gloria López

    2014-01-01

    Full Text Available 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 calorimetry experiments. At protein concentrations ≤0.56 mg·mL−1 the calorimetric transition is reversible and the data were fitted to a non-two-state model and deconvoluted into six transitions, whereas at concentrations greater than 0.56 mg·mL−1 the calorimetric transition is irreversible with an exothermic contribution to the thermogram. The apparent Tm increased linearly with the scan rate according to first order inactivation kinetics. The effect of additives on the calorimetric transition of xylanase is dependent on their nature. The addition of sorbitol transforms reversible transitions into irreversible transitions while stabilizing the protein as the apparent Tm increases linearly with sorbitol concentration. d-Glucono-1,5-lactone, a noncompetitive inhibitor in xylanase kinetics, and soluble xylan change irreversible processes into reversible processes at high protein concentration.

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

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    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.

  8. 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.

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

    Directory of Open Access Journals (Sweden)

    T Haryati

    2010-03-01

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

  10. Assay Methods and Unit Definitions of Xylanase Activity%木聚糖酶酶活性测定方法及酶活性单位定义

    Institute of Scientific and Technical Information of China (English)

    王晓丹; 郭丽琼; 赵力超; 林俊芳

    2009-01-01

    The definition and determination of xylanase activity was unified in this article. Through summarizing and comparing the present knowledge on different procedure for the determination of xylanase activity,one unit of xylanase activity was defined as the quantity of enzyme required to liberate 1 (xmol of reducing sugar ( as xylose) per minute at the measure conditions . This DNS method of expressing xylanase activity is scientifically sound and we suggest that this method be utilized as the universal and standard method of determining xylanase activity.%为统一木聚糖酶酶活性的单位定义及测定方法进行了实验.通过对现有酶活性的单位定义及测定方法的比较分析,得出木聚糖酶的酶活性单位定义为:在特定条件下,每分钟水解木聚糖形成1μmol木糖(还原糖)所需酶量为1个酶活力单位(U),并且采用还原糖法中的DNS法作为测定木聚糖酶酶活性的方法,该法具有合理性和科学性,建议以此作为木聚糖酶酶活性测定及酶活性单位定义的统一方法.

  11. Research Progress on the Production of Xylanase by Microorganism Fermentation%木聚糖酶微生物发酵生产的研究进展

    Institute of Scientific and Technical Information of China (English)

    孙孝梅; 黄建忠

    2012-01-01

    Xylanase is one of the most important enzymes for industry application, which has a wide range of applica- tions, such as pulp and paper industry, food, feed, medicine and biotransformation. Microorganism fermentation is the major approach to produce Xylanase. This paper reviews the research progress on the aspects such as the breeding of high-yielding bacterial strain for Xylanase and the production of Xylanase by microorganism fermentation, and finally looks into the distance to the research directions and prospects on the production of Xylanase by microorganism fermentation.%木聚糖酶是一种重要的工业用酶制剂,广泛应用于造纸、食品、饲料、医药及生物转化等领域。木聚糖酶最主要的来源是通过微生物发酵而成。该文综述了木聚糖酶高产菌株的选育和微生物发酵生产等方面的研究进展,并展望了木聚糖酶微生物发酵生产的研究方向及前景。

  12. OPTIMIZATION OF CELLULASE-FREE XYLANASE PRODUCED BY A POTENTIAL THERMOALKALOPHILIC PAENIBACILLUS SP.N1 ISOLATED FROM HOT SPRINGS OF NORTHERN HIMALAYAS IN INDIA

    Directory of Open Access Journals (Sweden)

    Sanjeev Kumar Verma

    2012-08-01

    Full Text Available Hot spring bacteria are found a novel source of highly active xylanase enzyme with significant activity at high temperature. Among bacteria, Paenibacillus sp.N1 isolated from hot water spring of Manikaran, H.P., India showed highest 24.60 IU.ml-1 of cellulase-free xylanase on Reese medium. Growth conditions including medium, incubation time, pH, temperature, inoculum size, aminoacids, carbon sources, nitrogen sources and additives that affect the xylanase production by Paenibacillus sp.N1 were studied sequentially using the classical “change-one factor at a time” method. The optimal cultivation conditions predicated from canonical analysis of this model were achieved by using basal salt medium on 3rd day, pH 9.0, temperature 50ºC with inoculum size of 12.5%, phenylalanine as aminoacid, xylose as carbon source, (NH42HPO4 as nitrogen source and Tween 20 as detergent added with an approximate yield of 52.30 IU.ml-1 escalating the over level of xylanase production by 113.38%. A rare combination of all characters i.e. thermoalkalophilic nature and high units of cellulase-free xylanase produced from a new Paenibacillus sp.N1 make it of special industrial interest.

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

    Science.gov (United States)

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

    2010-11-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

  15. High-yield recombinant xylanase production by Aspergillus nidulans under pyridoxine limitation.

    Science.gov (United States)

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

    2014-10-01

    The present study investigated the limitation of pyridoxine on an Aspergillus nidulans culture that produces xylanase B (XynB) as a client enzyme and was unable to synthesize pyridoxine. This technique was used to limit cell growth and divert substrate to product formation for a surface grown culture that could be used in trickle bed reactors. It was observed that growth was limited when pyridoxine was absent, while enzyme production was unaffected. Enzyme production was 1,026 U after 480 h of continuous fermentation, which was similar to a culture that grew on medium with pyridoxine. Furthermore, the present study investigated the growth rate of A. nidulans with pyridoxine in the medium and determined the productivity of XynB production with and without pyridoxine. A maximum growth rate of 0.311/h was observed. The maximum XynB productivity of 21.14 U/g h was achieved when pyridoxine was not added to the medium.

  16. Ethanol/Water Pulps From Sugar Cane Straw and Their Biobleaching With Xylanase from Bacillus pumilus

    Science.gov (United States)

    Moriya, Regina Y.; Gonçalves, Adilson R.; Duarte, Marta C. T.

    The influence of independent variables (temperature and time) on the cooking of sugar cane straw with ethanol/water mixtures was studied to determine operating conditions that obtain pulp with high cellulose contents and a low lignin content. An experimental 22 design was applied for temperatures of 185 and 215°C, and time of 1 and 2.5 h with the ethanol/water mixture concentration and constant straw-to-solvent ratio. The system was scaled-up at 200°C cooking temperature for 2 h with 50% ethanol-water concentration, and 1∶10 (w/v) straw-to-solvent ratio to obtain a pulp with 3.14 cP viscosity, 58.09 kappa-number, and the chemical composition of the pulps were 3.2% pentosan and 31.5% lignin. Xylanase from Bacillus pumilus was then applied at a loading of 5-150 IU/g dry pulp in the sugar cane straw ethanol/water pulp at 50°C for 2 and 20 h. To ethanol/water pulps, the best enzyme dosage was found to be 20 IU/g dry pulp at 20 h, and a high enzyme dosage of 150 IU/g dry pulp did not decrease the kappa-number of the pulp.

  17. 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.

  18. 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.

  19. 球毛壳ND35菌株在宿主植物上的侵染定殖%Infection and colonization of Chaetomium globosum ND35 on host plant

    Institute of Scientific and Technical Information of China (English)

    米士伟; 戴杨; 刘晓光; 孟庆果; 高克祥; Kurt Mendgen

    2011-01-01

    为了解球毛壳Chaetomium globosum ND35菌株在宿主植物上的侵染定殖方式和途径,以毛白杨组培苗为宿主植物,借助光学显微镜、扫描电镜、透射电镜,结合免疫荧光标记技术,研究了球毛壳ND35菌株子囊孢子萌发后在毛白杨上的侵染行为及其菌丝在组培苗根部的定殖。结果显示,子囊孢子萌发后形成的菌丝,能从杨树苗根、茎部表面细胞间的缝隙侵入或在根表面形成附着胞,进而形成侵染钉直接从表皮细胞侵入,在叶部主要从气孔侵入叶片内部。侵入根部的菌丝主要定殖于表皮细胞、外皮层细胞和细胞间隙,未进入内皮层和维管束组织。%Chinese white poplar plantlets from tissue culture were served as host plant in order to understand the mode and approach of infection and colonization of Chaetomium globosum ND35 on host plant.Infection of C.globosum ND35 ascospores on Chinese white poplar plantlets after germinating and colonization of C.globosum ND35 hyphae in poplar root were investigated by means of light microscopy,scanning electron microscopy,transmission electron microscopy and with immunofluorescent labeling.The results showed that hyphae of C.globosum ND35 invaded poplar root and stem from gap of epidermal surface cells,or directly penetrated epidermis by penetration peg formed from appressorium after ascospores germinating;and invaded poplar leaves mainly from stomata.Hyphae invaded root of poplar mainly colonized in the epidemic cells,the outermost cortical layer cells or intercellular spaces of cortical layer which was not found in endodermis and vascular tissues.

  20. Exogenous dietary xylanase ameliorates viscosity-induced anti-nutritional effects in wheat-based diets for White Pekin ducks (Anas platyrinchos domesticus).

    Science.gov (United States)

    Adeola, Olayiwola; Bedford, Michael R

    2004-07-01

    Nutrient utilisation and growth performance responses of White Pekin ducks (Anas platyrinchos domesticus) offered diets containing low- or high-viscosity wheat supplemented with xylanase were investigated in two studies. In Expt 1, six diets consisting of low-viscosity wheat or high-viscosity wheat supplemented with 0.0, 1.5 or 3.0 g xylanase (2590 units/g)/kg diet were used in a true metabolisable energy (TME) bioassay with eight 8-week-old ducks per diet group. In Expt 2, eight pens of ten 3-d-old ducks per pen for each of six wheat-based diets arranged in a 2 x 3 factorial of low-viscosity or high-viscosity wheat and 0.0, 1.5 or 3.0 g xylanase/kg were used in a 42 d growth study. High-viscosity wheat depressed (Pducks.

  1. 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

    2016-04-04

    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.

  2. 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.

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

    Science.gov (United States)

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

    2011-10-01

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

  4. Cellulose and hemicellulose-degrading enzymes in Fusarium commune transcriptome and functional characterization of three identified xylanases.

    Science.gov (United States)

    Huang, Yuhong; Busk, Peter Kamp; Lange, Lene

    2015-06-01

    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 decomposing enzymes (GH3, GH5, GH6, GH7, GH9, GH45 and AA9), and abundant hemicellulases. We further applied peptide pattern recognition to reveal nine and seven subfamilies of GH10 and GH11 family enzymes, respectively. The uncharacterized XYL10A, XYL10B and XYL11 enzymes of F. commune were classified, respectively, into GH10 subfamily 1, subfamily 3 and GH11 subfamily 1. These xylanases were successfully expressed in the PichiaPink™ system with the following properties: the purified recombinant XYL10A had interesting high specific activity; XYL10B was active at alkaline conditions with both endo-1,4-β-d-xylanase and β-xylosidase activities; and XYL11 was a true xylanase characterized by high substrate specificity. These results indicate that F. commune with genetic modification is a promising source of enzymes for the decomposition of lignocellulosic biomass.

  5. Simultaneous bioethanol distillery wastewater treatment and xylanase production by the phyllosphere yeast Pseudozyma antarctica GB-4(0).

    Science.gov (United States)

    Watanabe, Takashi; Suzuki, Ken; Sato, Ikuo; Morita, Tomotake; Koike, Hideaki; Shinozaki, Yukiko; Ueda, Hirokazu; Koitabashi, Motoo; Kitamoto, Hiroko K

    2015-12-01

    Bioethanol production using lignocellulosic biomass generates lignocellulosic bioethanol distillery wastewater (LBDW) that contains a large amount of xylose, making it a potential inexpensive source of xylose for biomaterials production. The main goal of this study was the production of useful enzymes from LBDW during treatment of this wastewater. In this study, we found that xylose strongly induced two yeast strains, Pseudozyma antarctica T-34 and GB-4(0), to produce novel xylanases, PaXynT and PaXynG, respectively. The nucleotide sequence of PaXynT [accession No. DF196774 (GAC73192.1)], obtained from the genome database of strain T-34 using its N-terminal amino acid sequence, was 91% identical to that of PaXynG (accession No. AB901085), and the deduced amino acid sequence is 98% identical. The specific activities of the purified PaXynT and PaXynG were about 52 U/mg. The optimal pH and temperature for both enzymes' activities were 5.2 and 50°C, respectively. They hydrolyzed xylan to xylose and neither had β-xylosidase (EC 3.2.1.37) activity, indicating that they are endo-β-xylanases (EC 3.2.1.8). With these results, we expect that PaXyns can be employed in saccharizing lignocellulosic biomass materials for the production of useful products just like other endoxylanases. After 72 h of LBDW fed-batch cultivation using a jar-fermentor, strain GB-4(0) produced 17.3 U/ml (corresponding to about 0.3 g/l) of PaXynG and removed 63% of dissolved organic carbon and 87% of dissolved total phosphorus from LBDW. These results demonstrate the potential of P. antarctica for xylanase production during LBDW treatment.

  6. Conformation analysis of a surface loop that controls active site access in the GH11 xylanase A from Bacillus subtilis.

    Science.gov (United States)

    Vieira, Davi Serradella; Ward, Richard John

    2012-04-01

    Xylanases (EC 3.2.1.8 endo-1,4-glycosyl hydrolase) catalyze the hydrolysis of xylan, an abundant hemicellulose of plant cell walls. Access to the catalytic site of GH11 xylanases is regulated by movement of a short β-hairpin, the so-called thumb region, which can adopt open or closed conformations. A crystallographic study has shown that the D11F/R122D mutant of the GH11 xylanase A from Bacillus subtilis (BsXA) displays a stable "open" conformation, and here we report a molecular dynamics simulation study comparing this mutant with the native enzyme over a range of temperatures. The mutant open conformation was stable at 300 and 328 K, however it showed a transition to the closed state at 338 K. Analysis of dihedral angles identified thumb region residues Y113 and T123 as key hinge points which determine the open-closed transition at 338 K. Although the D11F/R122D mutations result in a reduction in local inter-intramolecular hydrogen bonding, the global energies of the open and closed conformations in the native enzyme are equivalent, suggesting that the two conformations are equally accessible. These results indicate that the thumb region shows a broader degree of energetically permissible conformations which regulate the access to the active site region. The R122D mutation contributes to the stability of the open conformation, but is not essential for thumb dynamics, i.e., the wild type enzyme can also adapt to the open conformation.

  7. 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 xylanase...

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

    Directory of Open Access Journals (Sweden)

    Saddler Jack N

    2011-10-01

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

  9. 球毛壳菌60S核糖体蛋白L10a基因克隆与特性分析%Cloning and Characterization Analysis of 60S Ribosomal Protein L10a Gene from Chaetomium globosum

    Institute of Scientific and Technical Information of China (English)

    刘志华; 杨谦

    2006-01-01

    用粗糙脉孢菌(Neurospora crassa)XP_322380和赤霉菌(Gibberella zeae)PH-1(EAA76971)的60S核糖体蛋白L10a基因(60S ribosomal protein L10a,RPL10a)蛋白序列对球毛壳菌(Chaetomium globosum)ESTs序列数据库进行tBlastn检索,获得了球毛壳菌RPL10a cDNA序列.cDNA序列长765 bp,开放阅读框654 bp,编码217个氨基酸组成的多肽,蛋白分子量为23.9 kD.BlastP分析表明该基因氨基酸序列与粗糙脉胞菌相似最高为89%;与玉蜀黍黑粉菌(Ustilago maydis)相似性最低为78%.cDNA序列及推测的氨基酸序列在GenBank登录(登录号分别为AY669070,AAT74578).

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

    Science.gov (United States)

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

    2017-01-01

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

  11. Engineering a high-performance, metagenomic-derived novel xylanase with improved soluble protein yield and thermostability.

    Science.gov (United States)

    Qian, Changli; Liu, Ning; Yan, Xing; Wang, Qian; Zhou, Zhihua; Wang, Qianfu

    2015-03-01

    The novel termite gut metagenomic-derived GH11 xylanase gene xyl7 was expressed in Escherichia coli BL21, and the purified XYL7 enzyme exhibited high specific activity (6340U/mg) and broad pH active range of 5.5-10.0. Directed evolution was employed to enhance the thermostability of XYL7; two mutants (XYL7-TC and XYL7-TS) showed a 250-fold increase in half-life at 55°C, with a 10°C increase in optimal temperature compared to that of wild-type XYL7. A truncated enzyme (XYL7-Tr3) acquired by protein engineering showed similar catalytic properties as the wild-type, with a tenfold increase in soluble protein yield by the mutant. The reducing sugar produced by XYL7-TC was about fourfold greater than that produced by their parents when incubated with xylan at 60°C for 4h. The engineered novel xylanase exhibited superior enzymatic performance and showed promise as an excellent candidate for industrial application due to its high specific activity, stability and soluble protein yield.

  12. 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.

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

    Science.gov (United States)

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

    2010-04-01

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

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

    Directory of Open Access Journals (Sweden)

    RODICA CHEREJI

    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.

  15. Characterizing and improving the thermostability of purified xylanase from Aspergillus niger DFR-5 grown on solid-state-medium

    Directory of Open Access Journals (Sweden)

    Ajay Pal

    2010-12-01

    Full Text Available  The thermostability of absolutely purified xylanase from Aspergillus niger DFR-5 was improved using polyols. Supplementation of sorbitol at 2M concentration was found to increase the half-life and D-value of xylanase at elevated temperatures (45-70ºC. Thermodynamic parameters associated with the process were analyzed revealing that the stability at higher temperatures was due to the increased enthalpy (∆Hº and free energy (∆Gº change of enzyme denaturation in the presence of sorbitol. The negative values of ∆Sº (-150.093 Jmol-1K-1 at 70ºC clearly indicated that enzyme underwent a significant process of aggregation during denaturation. The enzyme required divalent cations for maximum activity and inhibited by chelator. The diminution of activity by various thiol-binding agents and enhancement by reducing agents like β-ME confirmed the essentiality of cysteine for catalysis. The enzyme had a half-life and D-value of 277 and 921 days when stored at 4 ºC.

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

    Science.gov (United States)

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

    2011-09-01

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

  17. Engineering the hydrophobic residues of a GH11 xylanase impacts its adsorption onto lignin and its thermostability.

    Science.gov (United States)

    Rakotoarivonina, Harivony; Hermant, Béatrice; Aubry, Nathalie; Rémond, Caroline

    2015-12-01

    This study aimed to characterise the parameters governing the non-specific adsorption of a xylanase from Thermobacillus xylanilyticus (Tx-Xyn11) onto lignin isolated from maize stems. Such adsorption may be due to hydrophobic interactions between Tx-Xyn11 and lignin. Our strategy was to mutate hydrophobic residues present on the surface of Tx- Xyn11 into non-hydrophobic residues. Three mutants (P1, P2, and P3) with altered hydrophobic regions were produced and characterised. The thermostability of the P1 mutant was largely decreased compared with the thermostable Tx-Xyn11. The rate of adsorbed enzyme onto lignin was reduced to a similar extent for the P1 and P2 mutants, whereas the adsorption of the P3 mutant was less affected compared with that of Tx-Xyn11. When considered separately, the hydrophobic residues did not affect xylanase adsorption onto lignin. The addition of Tween 20 also led to the decreased adsorption of Tx-Xyn11 onto lignin. These results suggest that hydrophobic interactions are a key parameter in the interaction of Tx-Xyn11 with isolated lignin.

  18. Antioxidant capacity of arabinoxylan oligosaccharide fractions prepared from wheat aleurone using Trichoderma viride or Neocallimastix patriciarum xylanase.

    Science.gov (United States)

    Malunga, Lovemore Nkhata; Beta, Trust

    2015-01-15

    The effect of xylanase type (Trichoderma viride or Neocallimastix patriciarum) and graded ethanol fractionation on the antioxidant capacity (AOC) of arabinoxylan oligosaccharides (AXOS) obtained from wheat aleurone was investigated. AXOS yields were higher using N. patriciarum (62%) than T. viride (44%). The fraction (F100) collected at >80% ethanol concentration constituted 60% of total recovered AXOS. Degree of substitution ranged from 0.20 to 0.60 for ethanol graded fractions. Ferulic acid (FA) esterified to AXOS (8.0 μg/ mg) was 2-fold lower for the N. patriciarum treatment. The mean AOC (41.6, 183.1, and 394.9 μM TE/mg) of T. viride treated AXOS was >1.4-fold higher than N. patriciarum treatment using DPPH and ABTS and ORAC assays, respectively. Fraction F100 had highest AOC. AOC was influenced by the content of esterified FA (R(2)=0.94). The type of xylanase had a major influence on the AOC of the resultant AXOS rich in FA content.

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

    Science.gov (United States)

    Lærke, H N; Arent, S; Dalsgaard, S; Bach Knudsen, K E

    2015-09-01

    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 without (NX) or with addition of xylanase from Aspergillus niger (AN), (BS), or (TR) was fed to 8 ileal-cannulated barrows (initial BW 30.9 ± 0.3 kg) for 1 wk each according to a double 4 × 4 Latin square design. In Exp. 2, fine rye, fine wheat, and coarse wheat with or without a combination of xylanase from and were fed to 6 ileal-cannulated barrows (initial BW 33.6 ± 0.5 kg) for 1 wk according to a 6 × 6 Latin square design with a 2 × 3 factorial arrangement of enzyme and cereal matrix. Chromic oxide (0.2%) was used as an inert marker. Ileal effluent was collected for 8 h on d 5 and 7 and pooled for analysis. In Exp. 1, TR reduced intestinal viscosity of pigs fed rye from 9.3 mPa·s in the control diet (NX) to 6.0 mPa·s ( fat digestibility of fine wheat with enzyme addition ( < 0.012) in Exp. 2. Collectively, the results suggest that xylanase is more efficient in degrading arabinoxylan from wheat than from rye.

  20. Feed intake, growth, digestibility of dry matter and nitrogen in young pigs as affected by dietary cation-anion difference and supplementation of xylanase

    NARCIS (Netherlands)

    Dersjant-Li, Y.; Schulze, H.; Schrama, J.W.; Verreth, J.A.J.; Verstegen, M.W.A.

    2001-01-01

    An experiment was conducted to test the effect of dietary cation-anion difference (CAD, Na K -Cl, mEq/kg diet) and xylanase addition on feed consumption, digestibility of nutrients, plasma electrolyte balance and growth performance in young pigs. A 2 3 factorial arrangement with three dietary CAD le

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

    Science.gov (United States)

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

    2015-01-01

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

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

    NARCIS (Netherlands)

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

    2012-01-01

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

  3. A new chitinase-like xylanase inhibitor protein (XIP from coffee (Coffea arabica affects Soybean Asian rust (Phakopsora pachyrhizi spore germination

    Directory of Open Access Journals (Sweden)

    Mehta Angela

    2011-02-01

    Full Text Available Abstract Background Asian rust (Phakopsora pachyrhizi is a common disease in Brazilian soybean fields and it is difficult to control. To identify a biochemical candidate with potential to combat this disease, a new chitinase-like xylanase inhibitor protein (XIP from coffee (Coffea arabica (CaclXIP leaves was cloned into the pGAPZα-B vector for expression in Pichia pastoris. Results A cDNA encoding a chitinase-like xylanase inhibitor protein (XIP from coffee (Coffea arabica (CaclXIP, was isolated from leaves. The amino acid sequence predicts a (β/α8 topology common to Class III Chitinases (glycoside hydrolase family 18 proteins; GH18, and shares similarity with other GH18 members, although it lacks the glutamic acid residue essential for catalysis, which is replaced by glutamine. CaclXIP was expressed as a recombinant protein in Pichia pastoris. Enzymatic assay showed that purified recombinant CaclXIP had only residual chitinolytic activity. However, it inhibited xylanases from Acrophialophora nainiana by approx. 60% when present at 12:1 (w/w enzyme:inhibitor ratio. Additionally, CaclXIP at 1.5 μg/μL inhibited the germination of spores of Phakopsora pachyrhizi by 45%. Conclusions Our data suggests that CaclXIP belongs to a class of naturally inactive chitinases that have evolved to act in plant cell defence as xylanase inhibitors. Its role on inhibiting germination of fungal spores makes it an eligible candidate gene for the control of Asian rust.

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

    NARCIS (Netherlands)

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

    2011-01-01

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

  5. Improvement of the optimum pH of Aspergillus niger xylanase towards an alkaline pH by site-directed mutagenesis.

    Science.gov (United States)

    Li, Fei; Xie, Jingcong; Zhang, Xuesong; Zhao, Linguo

    2015-01-01

    In an attempt to shift the optimal pH of the xylanase B (XynB) from Aspergillus niger towards alkalinity, target mutation sites were selected by alignment between Aspergillus niger xylanase B and other xylanases that have alkalophilic pH optima that highlight charged residues in the eight-residues-longer loop in the alkalophilic xylanase. Multiple engineered XynB mutants were created by site-directed mutagenesis with substitutions Q164K and Q164K+D117N. The variant XynB-117 had the highest optimum pH (at 5.5), which corresponded to a basic 0.5 pH unit shift when compared with the wild-type enzyme. However, the optimal pH of the XynB- 164 mutation was not changed, similar to the wild type. These results suggest that the residues at positions 164 and 117 in the eight-residues-longer loop and the cleft's edge are important in determining the pH optima of XynB from Aspergillus niger.

  6. Expression of the C-terminal family 22 carbohydrate-binding module of xylanase 10B of Clostridium themocellum in tobacco plant

    NARCIS (Netherlands)

    Olawole, O.

    2009-01-01

    Carbohydrate-binding modules have been shown to alter plant cell wall structural architecture. Hence, they have the potential application of being used to engineer the plant to produce tailor-made natural fibers in the cell wall. The Clostridium thermocellum xylanase, Xyn10B, contains two CBMs that

  7. 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.

  8. 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 .

  9. 2株内生真菌对菊花抗旱特性的影响%Effect of PEG stress on plantlets of Chrysanthemum morifolium induced by endophytic Botrytis sp.(C1)and Chaetomium globosum(C4)

    Institute of Scientific and Technical Information of China (English)

    宋文玲; 刘晓珍; 蔡信之; 孙迪; 戴传超

    2011-01-01

    The effect of the endophytic fungi Botrytis sp. ( C1 ) or Chaetomium globosum (C4) on the drought resistance of Chrysanthemum morifolium was studied. Ch. morifolium plantlets were inoculated with C1, C4 and cultured in the pots for 60 days, then the plantlets were stressed by 0%, 10%, 20%, 30%, 40% PEG6000 respectively in order to simulate different drought conditions. Biomass, the activities of SOD, POD, PAL, the contents of MDA and soluble protein of each group were determined. The results showed that endophytic fungi groups grew better than the control ( without inoculation endophytic fungi). With the increasing of the concentration of PEG6000, the biomass of Ch. morifolium of each groups decreased, while the biomass of fungi groups was significantly higher than that of control, moreover C4 group higher than C1 group. With the concentration of PEG increasing, the content of MDA of each group increased too, while POD activity and soluble protein content of all treatments increased at first and then decreased. SOD activity and PAL activity of the control were increased with the increase of PEG concentration, but SOD activity of the two ftmgi groups were stable. After been stressed by different concentrations of PEG, MDA content of two fungi groups were always lower than the control, while SOD activity, POD activity, PAL activity and soluble protein content were higher. In conclusion, endophytic fungi can increase the drought resistance of Ch. morifolium.%目的:以PEG6000模拟干旱条件,研究接种内生真菌(葡萄孢属C1菌Botrytis sp.、球毛壳菌G4菌Chaetomium globosum对药用菊花Ch.morifolium抗旱性的影响.方法:分别用0%,10%,20%,30%,40%PEG6000胁迫菊花组培苗4 d,测定各处理组菊花生物量,叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)活性及叶片丙二醛(MDA)、可溶性蛋白含量.结果:模拟干旱胁迫后,接种内生真菌的菊花长势好于对照(未接菌),PEG6000

  10. 嗜热毛壳菌内切β-葡聚糖酶的分离纯化及特性%Purification and Properties of An Endocellulase from The Thermophilic Fungus Chaetomium thermophile

    Institute of Scientific and Technical Information of China (English)

    路梅; 李多川; 张成省

    2002-01-01

    探讨了液体发酵嗜热毛壳菌(Chaetomium thermophile)产生的内切β-葡聚糖酶的分离纯化及特性.粗酶液经硫酸铵分级沉淀、DEAE-Sepharose Fast Flow阴离子层析、Pheny1-Sepharose疏水层析、Sephacry1 S-100分子筛层析等步骤便可获得凝胶电泳均一的内切β-葡聚糖酶.经12.5%SDS-PAGE和凝胶过滤层析法分别测得所分离纯化酶蛋白的分子量约为67.8kD和69.8kD.该酶反应的最适温度和pH分别为60℃和4.0~4.5在pH5.0条件下,该酶在60℃下稳定;70℃保温1h后,仍保留30%的活性;在80℃的半衰期为25min.金属离子对内切β-葡聚糖酶的活性影响较大,其中Na+对酶有激活作用;Fe2+、Ag+、Cu2+、Ba2+、Zn2+等对酶有抑制作用.该酶对结晶纤维素没有水解能力.

  11. Biochemical and thermodynamic characteristics of thermo-alkali-stable xylanase from a novel polyextremophilic Bacillus halodurans TSEV1.

    Science.gov (United States)

    Kumar, Vikash; Satyanarayana, T

    2013-09-01

    The purified extracellular xylanase of polyextremophilic Bacillus halodurans TSEV1 has been visualized as a single band on SDS-PAGE and eluted as single peak by gel filtration, with a molecular mass of 40 kDa. The peptide finger print and cloned xylanase gene sequence analyses indicate that this enzyme belongs to GH family 10. The active site carboxyl residues are mainly involved in catalysis, while tryptophan residues are involved in substrate binding. The enzyme is optimally active at 80 °C and pH 9.0, and stable in the pH range of 7.0-12.0 with T 1/2 of 35 min at 80 °C (pH 9.0). Activation energy for birch wood xylan hydrolysis is 30.51 kJ mol(-1). The K m, V max and k cat (birchwood xylan) are 2.05 mg ml(-1), 333.33 μmol mg(-1 )min(-1) and 3.33 × 10(4) min(-1), respectively. The pKa1 and pKa2 of ionizable groups of the active site that influence V max are 8.51 and 11.0. The analysis of thermodynamic parameters for xylan hydrolysis suggests this as a spontaneous process. The enzyme is resistant to chemical denaturants like urea and guanidinium-HCl. The site-directed mutagenesis of catalytic glutamic acid residues (E196 and E301) resulted in a complete loss of activity. The birch wood xylan hydrolyzate contained xylobiose and xylotriose as the main products without any trace of xylose, and the enzyme hydrolyzes xylotetraose and xylopentaose rapidly to xylobiose. Thermo-alkali-stability, resistance to various chemical denaturants and mode of action make it a useful biocatalyst for generating xylo-oligosaccharides from agro-residues and bleaching of pulp in paper industries.

  12. Production and Accumulation of Xylooligosaccharides with Long Chains by Growing Culture and Xylanase of a Mutant Strain of Bacillus pumilus X-6-19

    Institute of Scientific and Technical Information of China (English)

    Qingzhu Yuan; Tsuyoshi Adachi; Shinji Takenaka; Shuichiro Murakami; Machiko Tanaka; Kenji Aoki

    2008-01-01

    Bacillus pumilus X-6-9 isolated from soil and subsequently identified, produced xylooligosacchatides with long chainsfrom xylan and accumulated them in the culture. By improving the culture conditions and mutating the bacterium, a 3.2-fold increasein the production of the xylooligosaccharides was established, when compared to the original culture conditions of B. pumilus X-6-19.The addition of D-glucose to the culture of the mutant swain U-3 of B. pumilus X-6-9 repressed the synthesis of β-xylosidase, but notxylanase. Thus, it was revealed that strain U-3 was a good organism for the production and accumulation of xylooligosaccharideswith long chains from xylan by a microbial culture. Xylanase produced by strain U-3 was purified to homogeneity and characterized.The hydrolyzates generated by the purified xylanase contained xylobiose, xylotrinse, xylotewaose, and xylopentaose, but not xylose.

  13. The N-terminal cellulose-binding domain of EGXA increases thermal stability of xylanase and changes its specific activities on different substrates

    Institute of Scientific and Technical Information of China (English)

    Ming Ding; Yigang Teng; Qiuyu Yin; Jie Zhao; Fukun Zhao

    2008-01-01

    A full-length EGXA enzyme from a mollusk, Ampullaria crossean, was cloned into pFastBac vector and then heterogeneously expressed in insect Tn5 cells. Its natural N-terminal signal peptide worked well in the insect Tn5 cells.The recombinant EGXA was a 63 kDa protein and had active endo-β-1,4-glucanase (EC 3.2.1.4) and endo-β-1,4-xylanase (EC 3.2.1.8). The specific activity of endo-β-1,4-xylanase was higher than in the EGX, which was purified from the stomach tissues of Ampullaria crossen. The N-terminal cellulosebinding domain of EGXA made it bind to cellulose and xylan more efficiently. This cellulose-binding domain also increased the thermal stability of this recombinant enzyme and decreased the recombinant EGXA's specific activities on p-nitrophenyi-β-D-cellobioside and sodium carboxymethyl cellulose.

  14. Individual and combined effects of water addition with xylanases and laccase on the loaf quality of composite wheat–cassava bread

    DEFF Research Database (Denmark)

    Serventi, Luca; Skibsted, Leif H.; Kidmose, Ulla

    2016-01-01

    hardness was measured at 58 % water addition, likely due to insufficient plasticization of the gluten–starch network, while hardening and crumb structure collapse were observed at 76 % water addition. Enzyme evaluation revealed higher pore size upon treatment with the xylanase Panzea® BG (Panzea) compared......The objective was to study how water addition and addition of enzymes like xylanases and a laccase will improve the loaf quality of composite wheat–cassava bread. The loaf quality was determined by sensory profiling, volume measurement and texture profile analysis. High intensity of sensory...... of polyphenols and/or arabinoxylans. In confirmation to these findings, the combination of high water addition (70 %) and Panzea treatment generated larger and better structured bread....

  15. 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 the......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...... of a thermophilic eukaryote for studying complex molecular machines....

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

    OpenAIRE

    Marli Camassola; Dillon, Aldo J.P.

    2014-01-01

    The main limitation to the industrial scale hydrolysis of cellulose is the cost of cellulase production. This study evaluated cellulase and xylanase enzyme production by the cellulolytic mutant Penicillium echinulatum 9A02S1 using pretreated sugar cane bagasse as a carbon source. Most cultures grown with pretreated bagasse showed similar enzymatic activities to or higher enzymatic activities than cultures grown with cellulose or untreated sugar cane bagasse. Higher filter paper activity (1.25...

  17. Ectoine-mediated protection of enzyme from the effect of pH and temperature stress: a study using Bacillus halodurans xylanase as a model.

    Science.gov (United States)

    Van-Thuoc, Doan; Hashim, Suhaila O; Hatti-Kaul, Rajni; Mamo, Gashaw

    2013-07-01

    Compatible solutes are small, soluble organic compounds that have the ability to stabilise proteins against various stress conditions. In this study, the protective effect of ectoines against pH stress is examined using a recombinant xylanase from Bacillus halodurans as a model. Ectoines improved the enzyme stability at low (4.5 and 5.0) and high pH (11 and 12); stabilisation effect of hydroxyectoine was superior to that of ectoine and trehalose. In the presence of hydroxyectoine, residual activity (after 10 h heating at 50 °C) increased from about 45 to 86 % at pH 5 and from 33 to 89 % at pH 12. When the xylanase was incubated at 65 °C for 5 h with 50 mM hydroxyectoine at pH 10, about 40 % of the original activity was retained while no residual activity was detected in the absence of additives or in the presence of ectoine or trehalose. The xylanase activity was slightly stimulated in the presence of 25 mM ectoines and then gradually decreased with increase in ectoines concentration. The thermal unfolding of the enzyme in the presence of the compatible solutes showed a modest increase in denaturation temperature but a larger increase in calorimetric enthalpy.

  18. 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)

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

    Directory of Open Access Journals (Sweden)

    Qing Qing

    2011-06-01

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

  20. [Cultivation of a novel cellulase/xylanase producer, Trichoderma longibrachiatum mutant TW 1-59-27: production of the enzyme preparation and the study of its properties].

    Science.gov (United States)

    Bekkarevicha, A O; Nemashkalov, V A; Koshelev, A V; Goryazchev, D A; Bubnova, T V; Matys, V Yu; Osipov, D O; Kondrat'eva, E G; Okunev, O N; Sinitsyn, A P

    2015-01-01

    As a result of gamma-mutagenesis of Trichoderma longibrachiatum TW1 and the subsequent selection of improved producers, a novel mutant strain, TW1-59-27, capable of efficiently secreting cellulase and xylanase was obtained. In a fed-batch cultivation, the new TW1-59-27 mutant was significantly more active compared with the original TW1 strain. For instance, the activities of cellulase (towards carboxymethylcellulose) and xylanase in the culture broth (CB) increased by 1.8 and two times, respectively, and the protein content increased by 1.47 times. The activity of these enzymes in the dry enzyme preparation derived from the CB of the TW1-59-27 mutant was 1.3-1.8 times higher than that in the preparation derived from the original TW1 strain. It was established that the cellulase from the enzyme preparation of the mutant strain demonstrated the maximum activity at 55-65 degrees C; it occurred in xylanase at 60 degrees C. The pH optima of these enzymes were pH 4.5-5.0 and pH 5.0-6.0, respectively. It was shown that the content of endoglucanases in the enzyme preparation increased from 7% to 13.5%; the effect is largely driven by the secretion of endoglucanase-1. An enzyme preparation with increased endoglucanase-1 content is promising for use as a feed additive in agriculture.

  1. An approach to the production of soluble protein from a fungal gene encoding an aggregation-prone xylanase in Escherichia coli.

    Science.gov (United States)

    Le, Yilin; Peng, Jingjing; Wu, Huawei; Sun, Jianzhong; Shao, Weilan

    2011-04-08

    The development of new procedures and protocols that allow researchers to obtain recombinant proteins is of fundamental importance in the biotechnology field. A strategy was explored to overcome inclusion-body formation observed when expressing an aggregation-prone fungal xylanase in Escherichia coli. pHsh is an expression plasmid that uses a synthetic heat-shock (Hsh) promoter, in which gene expression is regulated by an alternative sigma factor (σ(32)). A derivative of pHsh was constructed by fusing a signal peptide to xynA2 gene to facilitate export of the recombinant protein to the periplasm. The xylanase was produced in a soluble form. Three factors were essential to achieving such soluble expression of the xylanase: 1) the target gene was under the control of the Hsh promoter, 2) the gene product was exported into the periplasm, and 3) gene expression was induced by a temperature upshift. For the first time we report the expression of periplasmic proteins under the control of an Hsh promoter regulated by σ(32). One unique feature of this approach was that over 200 copies of the Hsh promoter in an E. coli cell significantly increased the concentration of σ(32). The growth inhibition of the recombinant cells corresponded to an increase in the levels of soluble periplasmic protein. Therefore, an alternative protocol was designed to induce gene expression from pHsh-ex to obtain high levels of active soluble enzymes.

  2. Study on xylanase production by Neurospora sitophila in liquid fermentation%好食脉孢霉液态发酵产木聚糖酶的研究

    Institute of Scientific and Technical Information of China (English)

    邓永平; 刘晓兰; 艾瑞波; 郑喜群; 任凭

    2013-01-01

    试验以豆渣和麸皮为主要原料,研究了好食脉孢霉菌株产木聚糖酶的液态发酵条件.以木聚糖酶的催化活力为指标,确定液态发酵培养基由麸皮、豆渣和蛋白胨组成,初始pH值5.0、培养温度28℃、摇床转速150 r/min,培养72 h后木聚糖酶活力可达到476 U/ml.%Okara and bran as the main raw material, the liquid fermentation conditions of xylanase production by Neurospora sitophila was studied in this paper. Using xylanase catalytic activity as an indicator, the compositions of the liquid fermentation medium which were bran, okara and peptone were determined. When the initial pH was 5.0, incubation temperature was 28 ℃, the shaking speed was 150 r/min and culture time was 72 h, xylanase activity was 476 U/ml.

  3. Interactive effects of phytase and xylanase supplementation with extractable salt-soluble protein content of corn in diets with adequate calcium and nonphytate phosphorus fed to broilers.

    Science.gov (United States)

    Gehring, C K; Bedford, M R; Dozier, W A

    2013-07-01

    The objective was to determine the effects of extractable salt-soluble protein content of corn (PS) and exogenous enzyme supplementation on N, starch, and energy digestibility in broilers fed diets adequate in Ca and nonphytate P. Broilers were randomly distributed into floor pens (6 replicate pens per treatment) with 28 birds per pen at 1 d of age. Treatments consisting of 4 sources of corn varying in PS (A, 58.1; B, 54.2; C, 53.7; and D, 30.6 mg of BSA equivalent values) with or without phytase (0 and 1,000 phytase units/kg) and xylanase (0 and 16,000 units of xylanase activity/kg) were randomly assigned to each pen. Different sources of corn were provided from 1 to 9 and 24 to 29 d of age. However, enzyme treatments were provided throughout the experiment. From 1 to 9 d of age, no interactions were observed. Apparent ileal N digestibility (AIND) and apparent ileal digestible energy (IDE) of diets with the lowest PS (based on corn D) were lower (P ≤ 0.05) than those of diets with a higher PS. Phytase increased (P ≤ 0.01) AIND and IDE by 5 and 16%, respectively, and xylanase exerted the opposite effect (P ≤ 0.03). From 24 to 29 d of age, phytase and xylanase in combination resulted in reduced (P ≤ 0.05) AIND of diets with a low PS (based on corn D) compared with the basal diet in broilers. Broilers fed diets with the highest or lowest PS (based on corn A or D) had lower (3-way interaction; P ≤ 0.05) IDE when phytase and xylanase were supplemented in combination compared with either enzyme alone. In conclusion, responses to exogenous enzyme supplementation are not constant and are influenced by the source of ingredients as well as the age of broilers. The magnitudes of the responses to phytase on nutrient and energy digestibility were greater at 9 compared with 29 d of age.

  4. 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.

  5. Production of xylooligosaccharides from the steam explosion liquor of corncobs coupled with enzymatic hydrolysis using a thermostable xylanase.

    Science.gov (United States)

    Teng, Chao; Yan, Qiaojuan; Jiang, Zhengqiang; Fan, Guangsen; Shi, Bo

    2010-10-01

    The production of xylooligosaccharides (XOs) from corncobs was studied using a two-stage process based on a steam explosion pretreatment followed by enzymatic hydrolysis. Corncobs with different chip sizes were subjected to steam explosion under different experimental conditions of temperature and time, namely 188-204 degrees C for 2.5-7.5 min. The results indicate that corncobs were optimally steam exploded at 196 degrees C for 5 min, resulting in hemicellulose recovery of 22.8%. Especially, corncobs with large chip sizes (approximately 100 mm) during steam explosion pretreatment were suitable to produce XOs. Furthermore, a thermostable xylanase from Paecilomyces themophila J18 was used to hydrolyze steam explosion liquor of corncobs (SELC) for the production of XOs. A maximum XOs yield of 28.6 g XOs/100 g xylan in corncobs was achieved and XOs syrup contained more than 90% of xylobiose and xylotriose when the hydrolysis was carried out under the optimized conditions (pH 7.0, 70 degrees C, 7.5 U mL(-1) and 2.5 h). These results suggest that the process might be effective in production of XOs for industrial applications.

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

    Science.gov (United States)

    Campos Antoniêto, Amanda Cristina; Ramos Pedersoli, Wellington; Dos Santos Castro, Lílian; da Silva Santos, Rodrigo; Cruz, Aline Helena da Silva; Nogueira, Karoline Maria Vieira; Silva-Rocha, Rafael; Rossi, Antonio; Silva, Roberto Nascimento

    2017-01-01

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

  7. Potential of thermo and alkali stable xylanases from Thielaviopsis basicola (MTCC-1467) in biobleaching of wood kraft pulp.

    Science.gov (United States)

    Goluguri, Baby Rani; Thulluri, Chiranjeevi; Cherupally, Madhu; Nidadavolu, Nagaraju; Achuthananda, Das; Mangamuri, Lakshmi Narasu; Addepally, Uma

    2012-08-01

    Thermo- and alkali-stable xylanases produced from Thielaviopsis basicola (MTCC-1467) on low-cost carbon source like rice straw were evaluated for their potential application in biobleaching of wood kraft pulp. Enzyme treatment at retention time of 240 min with 20 IU/gm of dried pulp resulted in ~85.2 % of reduction in kappa number. When compared to control, 110.8, 93, and 72.2 % of enhancement in brightness (percent International Organization of Standardization), whiteness, and fluorescence, respectively, were observed for enzyme-treated pulp. Spectroscopic analysis showed significant release of chromophoric compounds from enzyme-treated pulp. Furthermore, scanning electron microscope studies of unbleached and enzyme bleached pulp revealed the effectiveness of enzymatic treatment. The enzyme-treated pulp subjected to later stages of chemical bleaching resulted in 16 % decrease in chlorine consumption along with considerable reduction in chemical oxygen demand percentage (14.5 %) level of effluent. Various pulp properties like fiber length, fiber width, burst strength, burst index, tear strength, tear index, tensile strength, and breaking length were also significantly improved after enzyme treatment when compared to control.

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

    Science.gov (United States)

    Campos Antoniêto, Amanda Cristina; Ramos Pedersoli, Wellington; dos Santos Castro, Lílian; da Silva Santos, Rodrigo; Cruz, Aline Helena da Silva; Nogueira, Karoline Maria Vieira; Silva-Rocha, Rafael; Rossi, Antonio

    2017-01-01

    Microorganisms play a vital role in bioethanol production whose usage as fuel energy is increasing worldwide. The filamentous fungus Neurospora crassa synthesize and secrete the major enzymes involved in plant cell wall deconstruction. The production of cellulases and hemicellulases is known to be affected by the environmental pH; however, the regulatory mechanisms of this process are still poorly understood. In this study, we investigated the role of the pH regulator PAC-3 in N. crassa during their growth on sugarcane bagasse at different pH conditions. Our data indicate that secretion of cellulolytic enzymes is reduced in the mutant Δpac-3 at alkaline pH, whereas xylanases are positively regulated by PAC-3 in acidic (pH 5.0), neutral (pH 7.0), and alkaline (pH 10.0) medium. Gene expression profiles, evaluated by real-time qPCR, revealed that genes encoding cellulases and hemicellulases are also subject to PAC-3 control. Moreover, deletion of pac-3 affects the expression of transcription factor-encoding genes. Together, the results suggest that the regulation of holocellulase genes by PAC-3 can occur as directly as in indirect manner. Our study helps improve the understanding of holocellulolytic performance in response to PAC-3 and should thereby contribute to the better use of N. crassa in the biotechnology industry. PMID:28107376

  9. 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.

  10. 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.

  11. 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

  12. 纤维素和木聚糖复合诱导合成木聚糖酶的研究%STUDY ON THE PRODUCTION OF XYLANASES CO-INDUCED BY CELLULOSE-XYLAN

    Institute of Scientific and Technical Information of China (English)

    刘超纲; 勇强; 余世袁

    2001-01-01

    This paper deals with the inducing function of cellulose andcellulose-xylan mixture on xylanases production with Trichoderma reesei. It was found that xylanase could be induced by cellulose, whereas cellulose-xylan exhibited a kind of co-inducing function to xylanases, which could greatly improve xylanase activity. Compared to pure xylan(5 g/L) xylanase activity could be increased by 45% by using xylan (4 g/L)-pulp(1 g/L)mixture as carbon source. This result provides the theoretical basis for xylanase production with plant fibre raw materials rich in xylan as carbon source.%以里氏木霉(Trichodermareesei)为产酶菌,分别对纤维素、纤维素和木聚糖诱导产酶的功能进行了研究。研究发现,纤维素具有诱导木聚糖酶合成的功能;纤维素和木聚糖混合对木聚糖酶合成具有促进作用,可大幅度提高木聚糖酶活力。与纯木聚糖(5g/L)产酶相比,纯木聚糖(4g/L)和纸浆(1g/L)混合产酶木聚糖酶活可以提高45%。研究成果为采用富含木聚糖的植物纤维原料作碳源制备木聚糖酶提供了理论依据。

  13. Energy utilization and growth performance of chickens fed novel wheat inbred lines selected for different pentosan levels with and without xylanase supplementation.

    Science.gov (United States)

    Pirgozliev, V; Rose, S P; Pellny, T; Amerah, A M; Wickramasinghe, M; Ulker, M; Rakszegi, M; Bedo, Z; Shewry, P R; Lovegrove, A

    2015-02-01

    Different F5 recombinant inbred lines from the cross Yumai 34×Ukrainka were grown in replicated trials on a single site in one harvest year at Rothamsted Research. A total of 10 samples from those lines were harvested and used in a broiler experiment. Twenty nutritionally complete meal-form diets that had 630 g/kg of wheat with different amounts of pentosan, with and without exogenous xylanase supplementation, were used to compare broiler growth performance and determine apparent metabolizable energy corrected for N retention (AMEn). We examined the relationship between the nutritive value of the wheat samples and their chemical compositions and results of quality tests. The amounts of total and water soluble pentosans in wheat samples ranged from 36.7 to 48.0 g/kg DM, and 6.7 to 11.6 g/kg DM, respectively. The mean crude oil and protein contents of the wheat samples were 10.5 and 143.9 g/kg DM, respectively. The average determined value for the kinematic viscosity was 0.0018 mPa.s, and 2.1 mPa.s for the dynamic viscosity. The AMEn of the wheat-based diets had a maximum range of 0.47 MJ/kg DM within the ten wheat samples that were tested. Xylanase supplementation improved (Ppentosan content. There was a negative relationship between the total pentosan content in the wheat and broiler growth performance. An increase by 10 g of pentosan per kg of wheat reduced (Ppentosan content. Supplementary xylanase improved energy and nutrient availability of all wheat samples that was independent of differences in pentosan content.

  14. Expression of the C-terminal family 22 carbohydrate-binding module of xylanase 10B of Clostridium themocellum in tobacco plant

    OpenAIRE

    Olawole, O.

    2009-01-01

    Carbohydrate-binding modules have been shown to alter plant cell wall structural architecture. Hence, they have the potential application of being used to engineer the plant to produce tailor-made natural fibers in the cell wall. The Clostridium thermocellum xylanase, Xyn10B, contains two CBMs that belong to family 22 (CBM22). The C-terminal CBM22-2 of the glycoside hydrolase (GH) 10 had been characterized to interact with xylan, a major hemicellulosic component in the secondary cell wall of ...

  15. 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.

  16. Effect of different pretreatment of sugar cane bagasse on cellulase and xylanases production by the mutant Penicillium echinulatum 9A02S1 grown in submerged culture.

    Science.gov (United States)

    Camassola, Marli; Dillon, Aldo J P

    2014-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Marli Camassola

    2014-01-01

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

  18. 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.

  19. Conservation of XYN11A and XYN11B xylanase genes in Bipolaris sorghicola, Cochliobolus sativus, Cochliobolus heterostrophus, and Cochliobolus spicifer.

    Science.gov (United States)

    Emami, Kaveh; Hack, Ethan

    2002-10-01

    Two types of xylanase gene, XYN11A ( XYL1) and XYN11B ( XYL2), were amplified by PCR and partially sequenced in four phytopathogenic species of the ascomycete fungal genus Cochliobolus (anamorph genus Bipolaris). Three of the species, C. heterostrophus ( B. maydis), C. sativus ( B. sorokiniana), and Bipolaris sorghicola (no teleomorph known), are interrelated; the fourth, C. spicifer ( B. spicifera), was found, through analysis of the 5.8S RNA and internal transcribed spacer (ITS) sequences of its ribosomal DNA, to be more distantly related to the other three. Isolates from all four species contain orthologous XYN11A and XYN11B genes, but a set of laboratory strains of C. heterostrophus gave no product corresponding to the XYN11B gene. The patterns of evolution of the two xylanase genes and ribosomal DNA sequences are mutually consistent; the results indicate that the two genes were present in the common ancestor of all Cochliobolus species and are evolving independently of each other.

  20. Comparison of a xylanase and a complex of non starch polysaccharide-degrading enzymes with regard to performance and bacterial metabolism in weaned piglets.

    Science.gov (United States)

    Vahjen, Wilfried; Osswald, Tanja; Schäfer, Klaus; Simon, Ortwin

    2007-04-01

    Weaned piglets were fed a wheat based diet either non-supplemented or supplemented with a multi-enzyme preparation or a xylanase mono-enzyme preparation, respectively. Both enzyme preparations increased live weight gain nonsignificantly, but only animals of the xylanase group showed a trend (p = 0.076) for an improved feed conversion. Only precaecal digestibility of total amino acids was improved significantly when the mono-enzyme preparation was added. Improvements of digestibility of crude fat, crude protein and starch did not reach the significance level. Both enzyme preparations reduced jejunal viscosity, however viscosity in the colon was only reduced by the mono-enzyme preparation. Both enzymes significantly reduced Lactobacillus spp. cell numbers as well as bacterial metabolites in the stomach and showed similar nonsignificant modifications in jejunum contents except for acetate in the mono-enzyme group. Total jejunal bile acids were unchanged. Compared to the control, the ratio of the main conjugated to the main deconjugated bile acid was significantly higher in the mono-enzyme group. This study has shown that the mono- and multi-enzyme preparation can lead to improved performance in wheat based diets for piglets. Like in poultry, the main mode of action seems to be the reduction of small intestinal viscosity. However, the generation of fermentable carbohydrates by the multi-enzyme preparation may mask beneficial effects on performance due to the development of an active bile acid deconjugating microbiota in the small intestine.

  1. IMPROVEMENT OF TCF BLEACHING OF OLIVE TREE PRUNING RESIDUE PULP BY ADDITION OF A LACCASE AND/OR XYLANASE PRE-TREATMENT

    Directory of Open Access Journals (Sweden)

    Raquel Martín-Sampedro,

    2012-02-01

    Full Text Available This study aimed at assessing the biobleachability of soda pulps obtained from olive tree pruning residue. The enzymatic (LMS pre-treatment was applied prior to a simple totally chlorine free (TCF bleaching sequence, consisting of an alkaline extraction and a hydrogen peroxide stage. Additionally, the effect of adding xylanase jointly with or prior to LMS was evaluated. All of these enzymatic pre-treatments were associated with an enhancement of the bleaching sequence. The best results were found when both enzymes were applied in the same stage: lowest hydrogen peroxide consumption (63 percent; kappa number, 11.6; brightness, 46 percent ISO. The mechanical properties observed were similar to those reported by other authors who have studied pulps from olive tree pruning residue. Finally, bleached pulps were subjected to accelerated ageing in order to assess the evolution of brightness and colorimetric properties. Although biobleached pulps showed lower stability upon ageing, the best optical properties, even after ageing, were observed in pulps treated with both xylanase and laccase.

  2. 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.

  3. Immobilization of His-tagged recombinant xylanase from Penicillium occitanis on Nickel-chelate Eupergit C for increasing digestibility of poultry feed

    Science.gov (United States)

    Driss, Dorra; Driss, Zied; Chaari, Fatma; Chaabouni, Semia Ellouz

    2014-01-01

    Recombinant xylanase 2 from Penicillium occitanis expressed with an His-tag in Pichia pastoris, termed PoXyn2, was immobilized on nickel-chelate Eupergit C by covalent coupling reaction with a high immobilization yield up to 93.49%. Characterization of the immobilized PoXyn2 was further evaluated. The optimum pH was not affected by immobilization, but the immobilized PoXyn2 exhibited more acidic and large optimum pH range (pH 2.0–4.0) than that of the free PoXyn2 (pH 3.0). The free PoXyn2 had an optimum temperature of 50 °C, whereas that of the immobilized enzyme was shifted to 65 °C. Immobilization increased both pH stability and thermostability when compared with the free enzyme. Thermodynamically, increase in enthalpy and free energy change after covalent immobilization could be credited to the enhanced stability. Immobilized xylanase could be reused for 10 consecutive cycles retaining 60% of its initial activity. It was found to be effective in releasing reducing sugar from poultry feed. Immobilization on Eupergit C is important due to its mechanical resistance at high pH and temperature. Hence, considerable stability and reusability of bound enzyme may be advantageous for its industrial application. PMID:24932488

  4. Study on Production of Xylooligosaccharides from Wheat Bran with Xylanase%酶法制备麸皮中低聚木糖的研究

    Institute of Scientific and Technical Information of China (English)

    隋明; 荣加超; 张阳; 曲鹏; 祝玉松; 张丽萍

    2012-01-01

    wheat bran is treated by xylanase to produce xylooligosaccharides. The best optimised process of enzymatic hydrolysis were determined by means of test,which was that :the accession of the xylanase is 800 IU/g,wheatbran dosage is 12% , zymohydrolysis temperature is 55 ℃, zymohydrolysis time is 4h, Under this condition, the yield of sugar could be up to 61. 02% , the DP is 3. 01. The yield of xylooligosaccharides is 28.67%.%木聚糖酶酶解小麦麸皮中的木聚糖制备低聚木糖,试验确定了最佳的酶解工艺条件,即酶添加量为800 IU/g,麸皮用量为12%,酶解温度为55℃,酶解时间为4h,总糖得率为61.02%,DP值为3.01,低聚木糖的得率为28.67%.

  5. Modulation of inhibitory activity of xylanase - α-amylase inhibitor protein (XAIP: binding studies and crystal structure determination of XAIP- II from Scadoxus multiflorus at 1.2 Å resolution

    Directory of Open Access Journals (Sweden)

    Dey Sharmistha

    2010-11-01

    Full Text Available Abstract Background Plants produce a wide range of proteinaceous inhibitors to protect themselves against hydrolytic enzymes. Recently a novel protein XAIP belonging to a new sub-family (GH18C was reported to inhibit two structurally unrelated enzymes xylanase GH11 and α-amylase GH13. It was shown to inhibit xylanase GH11 with greater potency than that of α-amylase GH13. A new form of XAIP (XAIP-II that inhibits α-amylase GH13 with a greater potency than that of XAIP and xylanase GH11 with a lower potency than that of XAIP, has been identified in the extracts of underground bulbs of Scadoxus multiflorus. This kind of occurrence of isoforms of inhibitor proteins is a rare observation and offers new opportunities for understanding the principles of protein engineering by nature. Results In order to determine the structural basis of the enhanced potency of XAIP-II against α-amylase GH13 and its reduced potency against xylanase GH11 as compared to that of XAIP, we have purified XAIP-II to homogeneity and obtained its complete amino acid sequence using cloning procedure. It has been crystallized with 0.1 M ammonium sulphate as the precipitating agent and the three-dimensional structure has been determined at 1.2 Å resolution. The binding studies of XAIP-II with xylanase GH11 and α-amylase GH13 have been carried out with surface plasmon resonance (SPR. Conclusion The structure determination revealed that XAIP-II adopts the well known TIM barrel fold. The xylanase GH11 binding site in XAIP-II is formed mainly with loop α3-β3 (residues, 102 - 118 which has acquired a stereochemically less favorable conformation for binding to xylanase GH11 because of the addition of an extra residue, Ala105 and due to replacements of two important residues, His106 and Asn109 by Thr107 and Ser110. On the other hand, the α-amylase binding site, which consists of α-helices α6 (residues, 193 - 206, α7 (residues, 230 - 243 and loop β6-α6 (residues, 180 - 192

  6. 牦牛瘤胃元基因组文库中木聚糖酶基因的分析%Analysis of xylanases derived from the metagenomic BAC clone library of yak rumen

    Institute of Scientific and Technical Information of China (English)

    王敏; 陈富荣; 张山; 朱雅新; 东秀珠; 黄力; 田瑞华; 董志扬; 戴欣

    2011-01-01

    [ Objective ] The diversity of rumen microbial xylanases and their degradation characteristics were studied and the resources of new xylanases genes and enzymes were provided. [ Methods ] According to the result of the high-throughput sequencing of the rumen microbial metagenome bacterial artificial chromosome ( B AC ) clones library, the diversity of xylanase genes was analyzed and screened. Then one xylanase and its downstream xylosidase gene screened was cloned and expressed in Escherichia coli. The enzyme characterization of the recombinant xylanase and xylosidase and their synergistic effect were studied. [Results] All 14 xylanases screened from the library belong to GH10 family proteins. These xylanases shared the amino acid sequence similarity between 20. 5% and 91. 3% . Intriguingly, 7 xylanase genes in different contigs were found to be followed by xylosidase gene. The specific enzyme activity of the xylanase (Xyn32) was 1.98 U/mg and no ferulic acid esterase activity was detected. The specific enzyme activity of the coupled xylosidase (Xyl33) was 0.07 IU/mg, and xylosidase ( Xyl33 ) also displayed the activity of arabinofuranosidase. In addition, the in vitro experiment confirmed the synergistic effect between the coupled xylanase and xylosidase.%[目的]了解牦牛瘤胃微生物木聚糖酶多样性及其降解特征,为木聚糖降解提供新的基因资源.[方法]根据对已构建的瘤胃微生物元基因组细菌人工染色体( BAC)克隆文库高通量测序结果的注释,筛选其中编码木聚糖酶的基因并进行多样性分析;对其中一个木聚糖酶基因及其连锁的木糖苷酶基因进行克隆表达和酶学性质表征,分析其协同作用.[结果]共筛选到14个木聚糖酶基因,均编码GH1O家族木聚糖酶,其氨基酸序列之间的相似性为20.5% -91.3%;其中7个木聚糖酶基因所在的不同的DNA片段(contig)上存在木糖苷酶基因,编码的木糖苷酶属于GH43或GH3糖苷水解酶家

  7. Difference in Ligocellulose Degradation of Endophytic Chaetomium globosum Isolates and Related Genes Analysis%球毛壳菌降解天然木质纤维素能力差异及酶系基因分析

    Institute of Scientific and Technical Information of China (English)

    郝晓冉; 牛学良; 李强; 潘皎; 朱旭东

    2014-01-01

    利用奠定了基础。%Objective: The decomposition abilities of lignocellulosic materials by Chaetomium globosum NK102, NK103, NK104 and NK105, endophytes from different plants were evaluated. Methods: The cellulose utilizing ca-pability of the C.globosum isolates were tested on carboxymethylcellulose agar and cellulose-congo red agar. The lignin utilizing experiments were performed on Bavendamm plates, and the degradative activity was compared by measuring the respective zone of color change. The lignocellulases production potential of these isolates using mi-crocrystalline cellulose, the leaves of Populus sp. and wood powder as the sole carbon source in liquid fermenta-tion was assessed. In addition, secondary metabolites produced by the C.globosum isolates were detected after 12 days cultivation. In the sequenced genome of C.globosum CBS148.51, genes encoding enzymes involved in lignocel-luloses degrading were identified by sequence homology alignment. Results: C.globosum NK102, NK103, NK104 and NK105 formed clear zones when cultured on carboxymethylcellulose or cellulose-congo red agar. In addition, all four isolates exhibited biodegradation capabilities against lignin and the strong-to-weak sequence was NK 103, NK102, NK105 and NK104 by Bavendamm reaction. In liquid culture, all isolates secreted cellulases and laccase on agar containing microcrystalline cellulose, the leaves of Populus sp., and wood powder. The highest activity of cellulase(0.76 U/mL) was obtained by NK102 when cultured on wood powder medium supplemented with peptone. The highest activity of laccase was obtained by NK103 when cultured on the leaves of Populus sp. medium. Chae-toglobosin A(ChA) was detected in the cultue broth of all four isolates. The yield of ChA was affected by carbon biomass and the highest yield was obtained by NK104 when cultured on the leaves of Populus with a yield of 14.88 mg/L. In the sequenced genome of C.globosum CBS148.51, we defined 119 genes encoding enzymes in-volved in cellulose and

  8. 诱导食用真菌代谢生产木聚糖酶的研究%Screening of Xylanase-producing Edible Fungi by Xylan Induction

    Institute of Scientific and Technical Information of China (English)

    邢振楠; 臧海莲; 王丹丽; 范冬茹

    2014-01-01

    以木聚糖为诱导底物,对66株大型食用真菌进行液体发酵诱导培养,采用超薄层琼脂板扩散法和DNS测定法进行初筛、复筛,最终筛选出3株产木聚糖酶较高的菌株。检测单因素结合正交试验,最终确定黑木耳LK8、玉田平菇、香菇236经木聚糖诱导,产木聚糖酶的最佳培养时间、pH值和底物木聚糖浓度分别为20d、7.5和1.0g/100mL;25d、7.0和1.0g/100mL;25d、5.0和0.75g/100mL,最佳条件组合时的酶活分别为:600.6IU/mL,672.5IU/mL,345IU/mL。并且,培养液pH值是影响诱导效果的主要因素。%Xylan as the inducing substrate ,using ultrathin-layer agar plate diffusion technique and DNS assay and rescreening ,eventually screened 3 xylanase producing high strains from 66 edible fungus in liq-uid fermentation induced culture .Black fungus LK8 ,Yutian mushroom ,M ushroom 236 have better en-zyme production capacity ,detection of single factor combined with orthogonal experiment finalized train-ing time ,pH and xylan concentration as the optimal condition for xylanase production were 20 d ,7 .5 and 1 .0 g/100mL ;25 d ,7 .0 and 1 .0 g/100mL ;25 d ,5 .0 and 0 .75 g/100mL ,respectively .Xylanase activi-ty under optimum conditions were :600 .6 IU/mL ,672 .5 IU/mL ,345 IU/mL .The culture solution pH value is the main factor affecting the induction effect .

  9. 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.

  10. 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...... marker. Ileal effluent was collected for 8 h on d 5 and 7 and pooled for analysis. In Exp. 1, TR reduced intestinal viscosity of pigs fed rye from 9.3 mPa·s in the control diet (NX) to 6.0 mPa·s (P ... AID of arabinoxylan by 91% to 107% (P fat digestibility of fine wheat with enzyme addition (P

  11. Development of a novel ultrasound-assisted alkali pretreatment strategy for the production of bioethanol and xylanases from chili post harvest residue.

    Science.gov (United States)

    Sindhu, Raveendran; Binod, Parameswaran; Mathew, Anil Kuruvilla; Abraham, Amith; Gnansounou, Edgard; Ummalyma, Sabeela Beevi; Thomas, Leya; Pandey, Ashok

    2017-03-04

    A novel ultrasound-assisted alkali pretreatment strategy was developed which could effectively remove lignin and hemicelluloses and improve the sugar yield from chili post harvest residue. Operational parameters that affect the pretreatment efficiency were studied and optimized. Inhibitor analysis of the hydrolyzate revealed that major fermentation inhibitors like furfural, 5-hydroxymethyl furfural as well as organic acids like citric acid, succinic acid and propionic acid were absent. Hence fermentation can be carried out without detoxification of the hydrolyzate. Changes in structural properties of the biomass were studied in relation to the pretreatment process using Scanning Electron Microscopy (SEM) and the changes in chemical composition were also monitored. The biomass pretreated with the optimized novel method could yield 0.428g/g of reducing sugars upon enzymatic hydrolysis. The hydrolyzate obtained by this novel pretreatment strategy was found to be suitable for bioethanol and xylanase production.

  12. 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 enteritis-related lesions (d 17) were more severe (P 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.

  13. Optimization of biobleaching of paper pulp in an expanded bed bioreactor with immobilized alkali stable xylanase by using response surface methodology.

    Science.gov (United States)

    Senthilkumar, Sundar Rajan; Dempsey, Michael; Krishnan, Chandraraj; Gunasekaran, Paramasamy

    2008-11-01

    Purified alkali stable xylanase from Aspergillus fischeri was immobilized on polystyrene beads using diazotization method. An expanded bed bioreactor was developed with these immobilized beads to biobleach the paper pulp in continuous mode. Response surface methodology was applied to optimize the biobleaching conditions. Temperature (degrees C), flow rate of pulp (ml/min) and concentration of the pulp (%) were selected as variables in this study. Optimal conditions for biobleaching process were reaction temperature 60 degrees C, flow rate of 2 ml/min and 5% (w/v) of pulp. The kappa number reduced from 66 in the unbleached pulp to 20 (reduction of 87%). This system proves to be a better option for the conventional chlorine based pulp bleaching.

  14. Medium Optimization of Production of Xylanase by Solid State Fermentation from Brevibacillus borstelensis – MTCC 9874 Isolated from Soil Sample of Eastern Nepal

    Directory of Open Access Journals (Sweden)

    Budhathoki, U.

    2011-01-01

    Full Text Available The main aim of this study was to optimize production medium in solid state fermentation for production of xylanase using Brevibacillus borstelensis MTCC 9874. The organism was isolated from Morang district of Nepal and it was grown for 96 h in five different mineral salt solutions (MMS with rice husk and MSS-1 was selected as a medium for further study based on xylanolytic activity measured using DNS method. Plackett Burman design (Minitab 15.1 was done with six variables viz. dipotassium hydrogen phosphate, rice husk, sodium chloride, magnesium sulphate, sodium carbonate and calcium chloride. The result showed that dipotassium hydrogen phosphate and rice husk were significant factors for xylanase production (> 95% confidence levels. Full factorial Centre composite design (CCD was used to optimize the two significant factors. Response surface and contour plot were used to locate the optimal value of the two factors. There was 279.88% increase in xylanolytic activity after optimization of the medium. Study of effect of temperature on xylanolytic activity showed that maximum xylanolytic activity (6.58±1.1 IU/mL was found at 60 °C. Optimum pH was found to be 7.6 (Xylanolytic activity = 6.81±2.32 IU/mL. Thermal stability study showed that the enzyme has a good stability at 60 °C (95.62%. Lineweaver – Burk plot showed that the enzyme has Vmax and Km values 0.1075 µg/mL .min and 1427.63 µg/mL respectively.

  15. 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.

  16. Impact of combined β-glucanase and xylanase enzymes on growth performance, nutrients utilization and gut microbiota in broiler chickens fed corn or wheat-based diets.

    Science.gov (United States)

    Munyaka, P M; Nandha, N K; Kiarie, E; Nyachoti, C M; Khafipour, E

    2016-03-01

    The effects of a xylanase and β-glucanase (XB) blend (2,500 U of xylanase and 250 U of β-glucanase per kg of complete feed) on growth performance, nutrients utilization and digesta microbiota in broiler chickens were investigated. A total of 140 day-old male Ross 308 broiler chicks were randomly assigned to 7 replicate cages and fed experimental diets. Diets were based on either corn or wheat without or with supplemental XB. Performance was monitored weekly and excreta were collected from d 17 to 20 for nutrients digestibility and AMEn measurements. On d 21, jejunal contents were collected for viscosity determination whereas ileal and cecal contents were obtained for microbial analysis by Illumina sequencing. Microbial data were analyzed using QIIME and PLS-DA whilst other data were analyzed using SAS. Birds fed wheat diets had higher (P diet whilst birds fed XB had better BWG (4%) and FCR (7%) than birds fed non-XB diets. Birds fed wheat diet had higher (P diet. XB reduced (P diet (-31%) than in corn-based diet (-10%). Birds fed wheat-based diet with XB had higher (3.5%) starch digestibility than birds fed this diet without XB. Janthinobacterium was associated with non-XB corn-based diet, whereas Ruminococcus, Lachnospiraceae, Lactobacillaceae, Peptostreptococcaceae, Clostridiales, Acidovorax and Blautia were associated with XB corn-based diet in the ileum. A relatively similar microbiome clustering was observed in wheat-based treatments in the cecum. There were no significant (P ≥ 0.05) correlations between selected ileal or cecal bacterial taxa and AMEn. Diet impacted growth performance but XB was efficacious across diet types, implying that degradation of dietary fibrous components by feed enzymes may stimulate performance in young birds. Data provided significant insight on ileal and cecal microbial profile associated with the dietary types and XB; however their functional roles require further investigations.

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

    Directory of Open Access Journals (Sweden)

    Dongling Zhan

    2014-09-01

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

  18. STUDY OF THE XYLANASE IN COMMERCIAL POULTRY FOODS SUPPLIED TO RATS UNDER LABORATORY CONDITIONS ESTUDO DA XILANASE EM RAÇÃO COMERCIAL DE AVES FORNECIDA A RATOS SOB CONDIÇÕES DE LABORATÓRIO

    Directory of Open Access Journals (Sweden)

    Celso de Paula Costa

    2007-09-01

    Full Text Available

    Two trials were conducted at Laboratório de Enzimologia do Instituto de Ciências Biológicas (I.C.B. and Departamento de Zootecnia da Escola de Agronomia e Veterinária da Universidade Federal de Goiás, to determine: a Influence of xylanase (fungic preparations in poultry rations fed to rats; b Possibility to use xylanase to improve commercial rations to monogastrics. A total of 48 rats were randomized in 12 metallic cages in an experimental design with 4 treatments and 3 repetitions. The treatments used were: Trial 1 : T - Commercial ration to broilers (table II; A - Commercial ration + 0.02% of xylanase; B - Commercial ration + 0.04% of xylanase; C - Commercial ration + 0.06% of xylanase. Trial 2 - As trial 1, but the levels of xylanase were 0.1, 0.2 and 0.4% (treatment A, B and C, respectively. The following conclusions are presented: a Increasing levels of xylanase didn’t improve weight gains, feed consumption and feed gain; b Different levels of crude fiber and xylanase should be studied in other experiments.

    Dois experimentos foram conduzidos nos Laboratórios de Enzimologia dos Instituto de Ciências Biológicas (ICE e no Departamento de Zootecnia da Escola de Agronomia e Veterinária da Universidade Federal de Goiás, objetivando determinar: a a influência da xilanase em rações de aves fornecidas a ratos; b a possibilidade de se empregar a xilanase em rações comerciais de monogástricos para o melhor aproveitamento de alimentos. Um total de 48 ratos albinos foram distribuídos ao acaso, em 12 gaiolas metálicas de fundo telado. O controle das dietas foi feito diariamente e as pesagens individuais foram feitas no início e final das duas semanas experimentais. O delineamento empregado foi o inteiramente casualizado, com 4 tratamentos e 3 repetições. Os tratamentos utilizados foram os seguintes

  19. 高产木聚糖酶的紫色红曲霉菌株筛选及酶学性质研究%Selection of High Xylanase Yielding Monascus pupureus Bacterium Strain and Its Properties of Enzyme

    Institute of Scientific and Technical Information of China (English)

    薛英丽; 邬应龙

    2011-01-01

    [目的]筛选紫色红曲霉木聚糖酶,并研究其酶学性质.[方法]采用平板筛选和固体培养方法获得了产木聚糖酶较高的紫红曲霉(Monascus pupureus)533.采用50%~70%硫酸铵盐析、Sephadex G-25凝胶层析、DEAE Sepharose fast flow阴离子交换层析和Sephacryl S-200凝胶层析进行纯化.最后,研究温度、pH、金属离子和化学物质对酶活性的影响.[结果]经分离纯化后得到相对分子质量约为46.0 kD的木聚糖酶.木聚糖酶最适反应温度为47℃;最适反应pH为5.5,pH稳定范围为4.5~5.5 ; Ca2+、Fe2+、Na+和Tween 80对酶活力有激活作用;Mn2+、Tris、EDTA、尿素和SDS对酶活力有抑制作用;Cu2+和K+对酶活力影响不大.[结论]该研究筛选到1株高产木聚糖酶的紫色红曲菌株,扩大了产木聚糖酶的菌株范围.%[Objective J The research aimed to screen the xylanase from Monascus pupweus bacterium strain, and studied its properties of enzyme. [ Method] Monascus pupureus 533 was isolated from Monascus as the xylanase producer by the solid state culture. The xylanase was purified by 50% - 70% ammonium sulfate precipitation, Sephadex G-25, DEAE-Sepharose fast flow column chromatographies and Sephacryl S-200. Finally, the effects of reaction temperature, pH, metal ions and chemical matters on the enzyme activity were studied. [Result] The molecular weight of xylanase was 46.0 kD by separation and purification. The optimal temperature and pH for xylanase activity were 47 ℃ and 5. 5, respectively. The xylanase was stable when pH was 4.5 -5.5. Its activity was activated by Ca2+ , Fe2+ , Na+ and Tween80, inhibited by Mn2* , Tris, EDTA, urea and SDS, while Cu2+ and K+ had a little effect on its activity. [Conclusion] The study selects a high xylanase yielding Monascus pupureus bacterium strain, to enlarge the range of strain producing xylanase.

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

    Science.gov (United States)

    Gottschalk, Leda Maria Fortes; de Sousa Paredes, Raquel; Teixeira, Ricardo Sposina Sobral; da Silva, Ayla Sant’Ana; da Silva Bon, Elba Pinto

    2013-01-01

    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 °C 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. PMID:24294256

  1. 一株产木聚糖酶嗜热菌的鉴定及酶学性质%Identification of a thermophilic bacterium and preliminary characterization of the secreted xylanase

    Institute of Scientific and Technical Information of China (English)

    陈学敏; 刘培培; 张波

    2011-01-01

    从云南腾冲热泉水样中分离筛选得到一株产木聚糖酶的菌株.通过细菌形态观察、生理生化特性并结合16S rDNA序列分析,经鉴定,该菌为地芽孢杆菌(Geobacillus sp.),命名为Geobacillus sp.PZH1.对该菌株所产嗜热木聚糖酶及酶学特性进行了初步研究.SDS-PAGE和酶谱分析测得该酶分子量约为69 kD;酶的最适反应pH和最适反应温度分别为7.0和70℃,在pH 5.0-11.0和40℃-100℃范围内均有较高酶活;该酶在pH 5.0-12.0范围内和70℃以内具有较高的稳定性:40℃-100℃范围内,该木聚糖酶没有被检测到纤维素酶活性.%The bacterium isolated from a water sample of Yunnan tengchong hot spring can secrete a kind of thermophilic xylanase. It was identified and named as Geobacillus sp. PZH1 by morphologic observation, physio-biochemical characteristics and 16S rDNA sequence alignment. Subsequently, its secreted xylanase and the xylanase's characteristics were researched preliminarily. SDS-PAGE electrophoresis and zymogram analysis suggested that the xylanase's molecular mass was 69 kD; the optimum pH and temperature of the partially purified enzyme were 7.0 and 70 ℃ respectively, and it performed noted activities from pH 5.0 to pH 11.0 and from 40 ℃ to 100 ℃; it had high stability from pH 5.0 to pH 12.0 and under 70 ℃; from 40 ℃ to 100 ℃, no cellulase activity was detected for the partially purified xylanase.

  2. 维生素C复合木聚糖酶对馒头形态影响%Effect of compound with vitamin C and xylanase on the shape properties of Chinese steaming bread

    Institute of Scientific and Technical Information of China (English)

    张晶; 赵阳; 陈海华; 张滢滢; 王雨生

    2014-01-01

    Effect of compound with vitamin C and xylanase on the shape properties of Chinese Steaming bread was investigated based on the single factor experiment and orthodoxy test. The rheological properties of the dough with Vitamin C and xylanase were also studied by farinograph and extensograph. The results showed that the optimal shape properties of Chinese steaming bread was achieved with xylanase of 10~20 mg/kg or vitamin C of 8 mg/kg, respectively. The results of orthogonal test showed that the optimum formula of the compound was Vitamin C of 8 mg/kg and xylanase of 10 mg/kg. Under the above conditions, the specific volume and aspect ratio of Chinese steaming bread were 2.434 and 0.648, respectively. Compared with the control and the dough added of vitamin C or xylanase only, the processing qualities of the dough prepared by the optimum formula were effectively improved, which was in accordance with the results of shape properties.%该实验通过单因素及正交试验探讨了维生素C复合木聚糖酶对馒头形态影响,并采用粉质仪、拉伸仪对单独添加木聚糖酶、Vc及二者复合最优添加量下面团的流变性质进行测定。结果表明:木聚糖酶、Vc单独添加时,最适添加量分别为10~20 mg/kg、8 mg/kg。二者复合添加时最优组合为,木聚糖酶添加量10 mg/kg、Vc添加量8 mg/kg;在此条件下制得馒头比容为2.434,高径比为0.648。与空白对照及单独添加木聚糖酶或Vc的面团相比,木聚糖酶与Vc复合添加时面团加工品质较好,流动性与硬度之间达到平衡,这与馒头形态测定结果一致。

  3. 组合型木聚糖酶对21日龄黄羽肉鸡器官指数、内源消化酶活性以及肠道吸收功能的影响%Effect of combination xylanase on organ index,endogenous digestive enzyme activity and absorptive function of intestinal tract for yellow-feather broilers

    Institute of Scientific and Technical Information of China (English)

    徐昌领; 左建军; 冯定远

    2011-01-01

    This study was conducted to evaluate effects on animal organ and the difference between single xylanase and combinative xylanase by measuring organ index, secretion level of pepsin, trypsin,lipase and amylase and xylose in serum of 21-day Yellow-feather broiler.The indexes of liver, gizzard, glandular stomach, duodenum, jejunum and ileum were decreased and the pancreas index was increased with xylanase addtion in wheat-soybean meal diet.Moreover,the secretion levels of pepsin and trypsin were enhanced with xylanase supplementation.In addition, small intestine index was lowered with xylanase cocktail addtion than single xylanase.The secretion of lipase was enhanced with single xylanase supplementation than combinative xylanase.However, no difference was observed.The level of xylose in serum was increased when corn was replaced by wheat as engergy feed,which was lowered with xylanase supplementation.There was no difference among different treatments.%通过对21日龄黄羽肉鸡器官指数和胃蛋白酶、胰蛋白酶、胰脂肪酶、胰淀粉酶以及血清中木糖水平进行检测,研究酶制剂对动物内脏器官所产生的影响,以及组合酶和单酶之间的差异.木聚糖酶在小麦型日粮中添加,降低了肝脏、肌胃、腺胃、十二指肠、空肠和回肠的器官指数,增加了胰腺的器官指数;降低了胃蛋白酶和胰蛋白酶的分泌水平;组合酶比单酶降低小肠器官指数更明显,单酶增加了胰脂肪酶的分泌水平,组合酶降低了胰脂肪酶的分泌水平,差异不显著(P>0.05).小麦型日粮饲喂组血清中木糖的水平高于玉米型日粮组和加酶组,酶制剂的添加降低了血液中的木糖水平,各处理组之间差异不显著(P>0.05).

  4. THUMB-LOOPS UP FOR CATALYSIS: A STRUCTURE/FUNCTION INVESTIGATION OF A FUNCTIONAL LOOP MOVEMENT IN A GH11 XYLANASE

    Directory of Open Access Journals (Sweden)

    Gabriel Paës

    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.

  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

    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 ...

  6. Synergistic degradation of arabinoxylan with alpha-L-arabinofuranosidase, xylanase and beta-xylosidase from soy sauce koji mold, Aspergillus oryzae, in high salt condition.

    Science.gov (United States)

    Hashimoto, Tadaaki; Nakata, Yoshiyuki

    2003-01-01

    This study addresses induction and some properties of alpha-L-arabinofuranosidase from a soy sauce koji mold, Aspergillus oryzae HL15, cultured on solid and liquid media. Alpha-L-arabinofuranosidase was induced by soybean polysaccharide and secreted into media on solid cultivation; the enzyme was associated with mycelium as a cell-wall-bound form in liquid cultivation. A major alpha-L-arabinofuranosidase, which was purified homogeneously on SDS-PAGE, showed highest activity in the presence of 10% of NaCl; also, somewhat higher activity was observed even in 25% NaCl than in the absence of NaCl. Arabinoxylan was synergistically degraded by xylanase, beta-xylosidase, and alpha-L-arabinofuranosidase from A. oryzae HL15 in the condition of imitative pH 5.0 and 15% NaCl concentration of the soy sauce moromi mash. These results suggest that arabinose and xylose, which are closely related to melanoidin formation, can be released by synergistic action of these enzymes in soy sauce moromi mash.

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

    Science.gov (United States)

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

    2010-05-05

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

  8. Role of N-linked glycosylation in the enzymatic properties of a thermophilic GH 10 xylanase from Aspergillus fumigatus expressed in Pichia pastoris

    Science.gov (United States)

    Chang, Xiaoyu; Xu, Bo; Bai, Yingguo; Luo, Huiying; Ma, Rui; Shi, Pengjun; Yao, Bin

    2017-01-01

    N-Glycosylation is a posttranslational modification commonly occurred in fungi and plays roles in a variety of enzyme functions. In this study, a xylanase (Af-XYNA) of glycoside hydrolase (GH) family 10 from Aspergillus fumigatus harboring three potential N-glycosylation sites (N87, N124 and N335) was heterologously produced in Pichia pastoris. The N-glycosylated Af-XYNA (WT) exhibited favorable temperature and pH optima (75°C and pH 5.0) and good thermostability (maintaining stable at 60°C). To reveal the role of N-glycosylation on Af-XYNA, the enzyme was deglycosylated by endo-β-N-acetylglucosaminidase H (DE) or modified by site-directed mutagenesis at N124 (N124T). The deglycosylated DE and mutant N124T showed narrower pH adaptation range, lower specific activity, and worse pH and thermal stability. Further thermodynamic analysis revealed that the enzyme with higher N-glycosylation degree was more thermostable. This study demonstrated that the effects of glycosylation at different degrees and sites were diverse, in which the glycan linked to N124 played a key role in pH and thermal stability of Af-XYNA. PMID:28187141

  9. Novel intracellular GH10 xylanase from Cohnella laeviribosi HY-21: biocatalytic properties and alterations of substrate specificities by site-directed mutagenesis of Trp residues.

    Science.gov (United States)

    Kim, Do Young; Han, Mi Kyoung; Oh, Hyun-Woo; Bae, Kyung Sook; Jeong, Tae-Sook; Kim, Sung Uk; Shin, Dong-Ha; Kim, In-Ho; Rhee, Young Ha; Son, Kwang-Hee; Park, Ho-Yong

    2010-11-01

    The novel intracellular GH10 xylanase (iXylC) gene (1023-bp) of Cohnella laeviribosi HY-21 encoded a protein consisting of 340 amino acids with a deduced molecular mass of 39,330Da and a calculated pI of 5.81. The primary structure of iXylC was 70% identical to that of Geobacillus sp. GH10 enzyme (GenBank accession number: EDV78425). Xylanolytic activity of the His-tagged iXylC overproduced in Escherichiacoli BL21 was stimulated by 2.2-fold in the presence of 0.5% non-ionic detergents. iXylC produced a mixture of xylooligosaccharides (xylobiose to xylooctaose) from xylotriose and xylotetraose used as the hydrolytic substrate. In addition, it exhibited considerable cleavage activities for p-nitrophenylxylopyranoside (PNP-xylopyranoside) and PNP-cellobioside, indicating that iXylC is a unique GH10 enzyme. The hydrolytic activity (57.8IUmL(-1)) of iXylC toward PNP-xylopyranoside increased to 8.3-fold by W217A and W315A mutations, while mutations of W133A, W295A, and W303A abolished the hydrolytic activity of the enzyme.

  10. 一株高产木聚糖酶真菌的筛选及酶学性质分析%Screening of xylanase-producing fungus and analysis of enzymatic properties

    Institute of Scientific and Technical Information of China (English)

    孙婕; 郁惠蕾; 李春秀; 寇振福; 许建和

    2013-01-01

    从近百份土样中,首先筛选得到具有明显半纤维素水解活性透明圈的菌株564株,其中202株的木聚糖酶活力用96孔板筛选方法进行了验证.结果显示:基于96孔深孔板测定的酶活力与初筛测得的透明圈活力趋势一致.最终筛选得到一株高产木聚糖酶的菌株ECU2023,经核糖体rDNA内部转录间隔区的序列对比,鉴定为泡盛曲霉(Aspergillus awamori).研究获得了其最佳培养条件:pH 6.0,培养时间4d,C源为30~ 40 g/L玉米芯粉.经过(NH4)2SO4沉淀和Superdex G-75凝胶过滤层析后,其中主要木聚糖酶成分的最适反应pH为6.0,最适反应温度为50℃.%Hundreds of various soil samples were collected. Firstly,564 strains with halo-forming zones were isolated. Then, the xylanase activities of 202 strains were further verified by a 96-deep-well microplate screening assay. The activities from 96-deep-well microplate and halo-forming plate showed the same trend. The best strain ECU2023 with the highest xylanase production was identified as Aspergillus awamori by internal transcnbed spacer (ITS) sequencing. The culture conditions were optimized as follows:30 ~ 40 g/L corn cob powder, pH 6. 0, cultivation for 4 d. Furthermore, the main xylanase component was crudely was purified by ammonium sulfate precipitation and Superdex G-75 chromatograph. The xylanase activity was optimally performed at 50 ℃ and pH 6.0.

  11. Studies on solid state fermentation of xylanase for cellulosic ethanol%纤维素乙醇木聚糖酶的固体发酵工艺研究

    Institute of Scientific and Technical Information of China (English)

    杨付伟; 王林风; 任建伟; 赵子高; 程远超

    2011-01-01

    该研究立足于河南天冠企业集团纤维素乙醇项目,以酶解糖化工艺对木聚糖酶的需求为出发点,利用黑曲霉X06作为产酶菌株,采用固体发酵工艺,通过正交设计试验,优化了培养基配方和发酵控制工艺,最优方案:麸皮:玉米芯=6:4、硝酸铵4%、尿素1%、磷酸二氢钾0.4%、硫酸镁0.2%、初始含水量65%、初始pH=4.0、温度28℃、环境相对湿度70%、72 h酶活达到10096.74 IU/g.这一方案在生产中得到进一步放大和优化,所生产的固体木聚糖酶应用在秸秆酶解工艺中,酶解液中木糖含量提高了66.9%.%Based on the needs of xylanase in the cellulosic ethanol project of Henan Tianguan Group Co., Ltd., Using Aspergillus niger X06 as xylanase-producing strain, through orthogonal experiment, the optimal solution of solid state fermentation was obtained as follows: the wheat bran to corn cob ratio is 6:4,the NH4NO3 content is 4%, the CO(NH2)2 content is 1%, the KH2PO4 content is 0.4%, the MgSO4·TH2O content is 0.2%, the initial moisture content is 65%, the initial pH value is 4.0, the incubation temperature is 28 ℃, the relative humidity of the environment is 70%. The activity of xylanase could reach 10 096.74 IU/g in 72 h. This program has been amplified and further optimized in large-scale production. The production of solid xylanase has been used in straw hydrolysis process, and the xylose content increased by 66.9%.

  12. 嗜热菌Geobacillus sp.PZH1产木聚糖酶发酵条件的优化%Optimization of fermentation conditions of xylanase from thermophilic bacterium Geobacillus sp.PZH1

    Institute of Scientific and Technical Information of China (English)

    刘培培; 陈学敏; 王石峰; 张波

    2012-01-01

    The culture conditions for alkali-thermo-stable xylanase production from the thermophilic bacterium Geobacillus sp.PZH1 were optimized.Five factors,such as carbon source,nitrogen source,initial pH,inoculum size and fermentation temperature,were researched in single-factor experiment.C/N ratio,initial pH and inoculum size were researched in orthogonal experiment.The results showed that the xylanase yield reached a highest level for 7d culture,and the best combination of fermentation conditions for alkali-thermo-stable xylanase production from the thermophilic bacterium Geobacillus sp.PZH1 was brichwood xylan as carbon source,beef extract as nitrogen source,C N ratio 2∶3,initial pH 7.0,inoculum size 4%,fermentation temperature 50℃ and fermentation time 7d.Under these optimal conditions,the xylanase production from the thermophilic bacterium Geobacillus sp.PZH1 was 2.56IU/mL,1.44 fold higher than that before the optimization.%对嗜热菌Geobacillus sp.PZH1发酵产嗜热耐碱木聚糖酶的培养条件进行了优化研究。对碳源、氮源、初始pH、接种量以及发酵温度五个因素进行了单因素实验,在此基础上对碳氮比、初始pH以及接种量进行了正交实验。结果表明,该菌株在发酵培养7d时有最大产酶量,Geobacillus sp.PZH1发酵产木聚糖酶最佳发酵条件为:桦木木聚糖为碳源,牛肉膏为氮源,碳氮比2∶3,初始pH7.0,接种量4%,发酵温度50℃,发酵时间7d。在最佳产酶条件下进行发酵,木聚糖酶活力可达2.56IU/mL,是未优化前酶活的1.44倍。

  13. Effect of exogenous xylanase on sensory quality and texture of fresh bread%外源木聚糖酶对面包感官品质和质地的影响

    Institute of Scientific and Technical Information of China (English)

    王军节; 努力扎提; 张怀予; 李贞子

    2011-01-01

    The correlation between sensory evaluation and texture index applied in analysis quality of bread and the effect of exogenous xylanase on sensory quality and texture were studied.The results showed that xylanase application at 0.25 g/kg can improve sensory quality of bread significantly.The sensory score of bread by xylanase treatment was 11.7% higher than that of control.The exogenous enzyme can also increase the springiness,chewiness,gumminess and adhesiveness while decrease the hardness and cohesiveness.The springiness and chewiness of the treated bread were 23.9% and 50.8% higher than that in the control individually.The results also indicated that the relationship between the elastic ability of sensory evaluation and the springiness of textural analysis was positively correlated.It is suggested that appropriate exogenous xylanase could be used to ameliorate bread quality,and texture profile analysis would be an ideal method applied to evaluate the quality of bread instead of sensory analysis.%试验研究了外源木聚糖酶添加对面包感官品质及质地指标的影响,并进一步分析感官评价与质地多面性分析之间关系。结果表明:0.25g/kg木聚糖酶能显著提高面包感官品质,处理组感官得分比对照高11.7%;木聚糖酶还能显著提高弹性、咀嚼性、胶黏性和黏附性而降低硬度和内聚性,其中处理组弹性和咀嚼性改良最为明显,分别比对照组高23.9%和50.8%;研究还表明,感官评价弹柔性与质构分析弹性之间成正相关关系。由此表明,适宜外源木聚糖酶可用于改良面包品质,且质构多面性分析将是替代感官评价用于面包品质分析的理想方法。

  14. The critical role of N- and C-terminal contact in protein stability and folding of a family 10 xylanase under extreme conditions.

    Directory of Open Access Journals (Sweden)

    Amit Bhardwaj

    Full Text Available BACKGROUND: Stabilization strategies adopted by proteins under extreme conditions are very complex and involve various kinds of interactions. Recent studies have shown that a large proportion of proteins have their N- and C-terminal elements in close contact and suggested they play a role in protein folding and stability. However, the biological significance of this contact remains elusive. METHODOLOGY: In the present study, we investigate the role of N- and C-terminal residue interaction using a family 10 xylanase (BSX with a TIM-barrel structure that shows stability under high temperature, alkali pH, and protease and SDS treatment. Based on crystal structure, an aromatic cluster was identified that involves Phe4, Trp6 and Tyr343 holding the N- and C-terminus together; this is a unique and important feature of this protein that might be crucial for folding and stability under poly-extreme conditions. CONCLUSION: A series of mutants was created to disrupt this aromatic cluster formation and study the loss of stability and function under given conditions. While the deletions of Phe4 resulted in loss of stability, removal of Trp6 and Tyr343 affected in vivo folding and activity. Alanine substitution with Phe4, Trp6 and Tyr343 drastically decreased stability under all parameters studied. Importantly, substitution of Phe4 with Trp increased stability in SDS treatment. Mass spectrometry results of limited proteolysis further demonstrated that the Arg344 residue is highly susceptible to trypsin digestion in sensitive mutants such as DeltaF4, W6A and Y343A, suggesting again that disruption of the Phe4-Trp6-Tyr343 (F-W-Y cluster destabilizes the N- and C-terminal interaction. Our results underscore the importance of N- and C-terminal contact through aromatic interactions in protein folding and stability under extreme conditions, and these results may be useful to improve the stability of other proteins under suboptimal conditions.

  15. 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

  16. Feeding value of field beans (Vicia faba L. var. minor) with and without enzyme containing tannase, pectinase and xylanase activities for broilers.

    Science.gov (United States)

    Abdulla, Jalil Mahmwd; Rose, Stephen Paul; Mackenzie, Alexander Mackay; Pirgozliev, Vasil Radoslavov

    2017-04-01

    Effects of field beans with various tannin content and exogenous enzyme mixture containing tannase, pectinase and xylanase activities on N-corrected dietary apparent metabolisable energy (AMEn), coefficients of dry matter (DMR) and nitrogen retention (NR), fat digestibility, gastrointestinal tract (GIT) development, jejunal villus morphometry, ileal digesta viscosity and sialic acid were examined. Birds' growth performance and energy conversion ratio (ECR) were also measured. Birds were fed one of eight mash diets. The Control diet contained as major ingredients wheat (400 g/kg) and soybean meal (SBM) (127 g/kg and 221 g crude protein/kg and 12.83 MJ AMEn/kg. To reduce nutrient density, the Control diet also contained washed sand at 119 g/kg. Another three diets containing 300 g/kg of each of three experimental field bean cultivar samples in replacement for SBM and sand were also mixed. Each diet was fed to nine pens with two male Ross 308 broilers. Diets high in tannin had low AMEn, ECR, DMR and NR (p < 0.001). Feeding field beans increased (p < 0.001) the weights of the pancreas and the proventriculus and gizzard (PG) of the birds. Supplementing diets with the enzyme mixture improved (p < 0.001) feed conversion efficiency, AMEn and all nutrient utilisation coefficients despite the tannins in diets. The enzyme mixture reduced ileal digesta viscosity (p < 0.001) and the weight of pancreas, total GIT and PG (p < 0.05) of the birds. It can be concluded that the feeding value of field beans with different tannin contents may vary when fed to broilers. The supplementation of the enzyme mixture improved the feeding value of diets for broilers. The beneficial effect of the addition of the enzyme mixture seems to be mediated through reduced ileal digesta viscosity and improved nutrient availability.

  17. Effects of cellulase and xylanase enzymes mixed with increasing doses ofSalix babylonicaextract onin vitrorumengas production kinetics of a mixture of corn silage with concentrate

    Institute of Scientific and Technical Information of China (English)

    Abdelfattah Z M Salem; German Buenda-Rodrguez; Mona M M Elghandour; Mara A Mariezcurrena Berasain; Francisco J Pea Jimnez; Alberto B Pliego; Juan C V Chagoyn; Mara A Cerrillo; Miguel A Rodrguez

    2015-01-01

    Anin vitro gas production (GP) technique was used to investigate the effects of combining different doses ofSalix babylonicaextract (SB) with exogenous ifbrolytic enzymes (EZ) based on xylanase (X) and celulase (C), or their mixture (XC; 1:1 v/v) onin vitrofermentation characteristics of a total mixed ration of corn silage and concentrate mixture (50:50, w/w)as substrate. Four levels of SB (0, 0.6, 1.2 and 1.8 mL g–1 dry matter (DM))andfour supplemental styles of EZ (1 µL g–1 DM; control (no enzymes), X, C and XC (1:1, v/v) were used in a 4×4 factorial arrangement.In vitro GP (mL g–1 DM) were recorded at 2, 4, 6, 8, 10, 12, 24, 36, 48 and 72 h of incubation. After 72 h, the incubation process was stopped and supernatant pH was determined, and then ifltered to determine dry matter degradability (DMD). Fermentation parameters, such as the 24 h gas yield (GY24),in vitro organic matter digestibility (OMD), metabolizable energy (ME), short chain fatty acid concentrations (SCFA), and microbial crude protein production (MCP) were also estimated. Results indicated that there was a SB´EZ interaction (P0.05) on OMD, pH, ME, GY24, SCFA and MP. The combination of SB with EZ increased (P<0.001) OMD, ME, SCFA, PF72 and GP24, whereas there was no impact on pH. It could be concluded that addition of SB extract, C, and X effectively improved thein vitro rumen fermentation, and the combination of enzyme with SB extract at the level of 1.2 mL g–1 was more effective than the other treatments.

  18. 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.

  19. 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.

  20. 重组多功能木聚糖酶酶解条件的优化%Optimization of enzymatic hydrolysis of recombinant multi-functional xylanase

    Institute of Scientific and Technical Information of China (English)

    赵力超; 王燕; 刘晓娟; 林俊芳

    2014-01-01

    重组多功能木聚糖酶(recombinant multi-functional xylanase,RMFXase)是以草菇为表达载体,具有多种酶活力的新型酶。为揭示该酶与底物的作用规律,同时为转基因酶的应用提供基础数据,该文利用重组多功能木聚糖酶水解南方特色的甘蔗加工废弃物生产低聚木糖,通过响应面法建立以重组多功能木聚糖酶质量浓度、底物质量浓度和酶解时间为控制因素的反应动力学方程组,从而推导得到酶解最佳工艺参数。试验并通过高效液相色谱法法,监测反应过程中产物的动态变化,分析产物的生成规律。结果表明,通过响应面法推导出符合连续式酶解反应器的最佳酶解工艺参数为:加酶量900 U/mL、底物质量浓度110 mg/mL、反应时间80 min。该条件下酶解木聚糖含量为54.7%的蔗渣半纤维素,得到的低聚木糖DP值达到3,酶解率达到23.94%。酶解过程中重组多功能木聚糖酶表现出高内切-β-1,4-木聚糖活力,优先降解大分子木聚糖,后期产物主要以木二糖和木三糖为主,且底物纯度越高,底物利用率越高。%The utilization of microorganisms or their enzymes may be the most promising method to produce Xylo-oligosaccharides (XOs) from agricultural wastes because of fewer undesirable by-products and fewer monosaccharides produced, and no special equipments required. Industrial-scale production of XOs using bioconversion requires low-cost substrates with a high xylan content, low lignin content and enzymes with high activity, process stability and β-1,4-xylosidic bond specificity. In the current study, bagasse, by-products of sugarcane production, was used as substrates for XOs production. And hydrolysis of bagasse hemicellulose was carried out with a recombinant multi-functional xylanase (RMFXase). RMFXase is a novel enzyme produced from genetically engineered straw mushroom (Volvariella volvacea) strain, which has exo

  1. Scientific Opinion on the safety and efficacy of Rovabio® Excel (endo-1,3(4-beta-glucanase and endo-1,4-beta-xylanase as a feed additive for lactating sows

    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 Rovabio® Excel is a preparation of endo-1,3(4-beta-glucanase and endo-1,4-beta-xylanase that is intended to be used as a feed additive for lactating sows, at a dose of 1 500 glucanase U and 1 100 xylanase U/kg feed, in order to minimise the mobilisation of body reserves of sows during lactation. The European Food Safety Authority issued an opinion on the safety and efficacy of Rovabio® Excel as a feed additive for chickens and turkeys for fattening, laying hens, piglets (weaned and pigs for fattening, ducks, guinea fowl, quails, geese, pheasants and pigeons. The full description of the formulations, manufacturing processes, purity, stability and homogeneity of the product is given in that assessment. The FEEDAP Panel considers that the safety aspects, other than for the new target species, are covered in the previous assessment and would not be affected by the extension of use requested. The results of a tolerance study showed that 200-fold the recommended dose was tolerated well by the sows when offered for a period of seven weeks, including gestation and lactation. Therefore, the Panel concludes that the recommended dose is safe in lactating sows. The Panel cannot conclude on the efficacy of Rovabio® Excel from the available efficacy data.

  2. 产木聚糖酶海洋微生物的筛选与诱变育种%Screening and breeding by induced mutation of xylanase-producing marine microorganism

    Institute of Scientific and Technical Information of China (English)

    黄小云; 林娟; 林小洪; 王国增; 叶秀云

    2015-01-01

    With self-made xylan as the only carbon source of primary screening medium , totally 60 strains which showed transparent zones and different morphology were screened from different marine samples .The shake flask fermentation results showed that 38 strains had xylanase-producing capabil-ities and strain B659 had the highest xylanase activity of 525.3 U· mL-1 .Via morphological charac-teristics and 16S rDNA sequence analysis , strain B659 was identified as Bacillus.The stable mutant strain G3-17 with 13.9%enhancement of xylanase activity was screened from the UV mutagenesis of strain B659.A stable mutant strain W1-40 with 11.6% xylanase activity increase was obtained by microwave mutation and screening from strain G 3-17.In 72 h fermentation of strain B659 and strain W1-40, the xylanase activity of latter (645.2 U· mL-1 ) achieved 24.6%elevation than the former (517.9 U· mL-1).%以自制木聚糖为初筛培养基的唯一碳源,从多个海洋来源样品中一共筛到60株有透明圈且形态各异的菌株,摇瓶发酵结果显示,38株具有产木聚糖酶能力,其中B659菌株产酶能力最高,酶活力为525.3 U· mL-1.结合B659菌株的形态特征和16S rDNA序列分析鉴定该菌株属于Bacillus属.对B659菌株进行紫外诱变,筛选得到酶活提高13.9%且稳定遗传的突变菌株G3-17;对G3-17菌株进一步进行微波诱变得到酶活较G3-17菌株高出11.6%且稳定遗传的突变菌株W1-40.对B659菌株和W1-40突变菌株进行发酵试验,72 h时W1-40菌株的酶活力达到645.2 U· mL-1,比B659菌株(517.9 U· mL-1)提高24.6%.

  3. 戊聚糖及木聚糖酶对肉雏鸡器官发育的影响%Effects of Pentosans and Xylanase Supplementation on Organ Development of Broiler Chick

    Institute of Scientific and Technical Information of China (English)

    盛清凯; 赵红波; 黄保华

    2013-01-01

    The experiment was conducted to study effects of water-soluble pentosan (WSP),alkai-extractable pentosan (AEP) and xylanane on the organ development of broiler chicks fed on the corn-soybean meal diets and probe into the difference of their mechanisms.Three hundred and fifty 1-day-old Avain broiler chicks were randomly divided into 7 treatments with 5 replicates and 10 broilers in each replicates,fed corn-soybean meal basal diet and experimental diets supplemented with 50 mg/kg WSP,100 mg/kg.WSP,50 mg/kg AEP,100 mg/kg AEP,3 mg/kg xylanase,6 mg/kg xylanase,respectively.The experiment lasted for 15 days.The results showed that compared with control group,WSP and AEP and xylanase increased significantly body weight,the weight of bursa of fabricius and gizzard and glandular stomach,the length of gut (P <0.05),but had no significant effect on lactobacillus and streptococcus and salmonella and coliform in recta chyme.WSP and AEP raised the viscosity and the xylanase decreased the viscosity with the two chime viscosities extremely significant different (P <0.01).50 mg/kg WSP supplementation promoted the development of broiler chicks best.The result indicated WSP and AEP had different mechanism with xylanase in promoting the organ development of broiler chicks.%研究水溶戊聚糖(WSP)、碱溶戊聚糖(AEP)和木聚糖酶对玉米-豆粕型日粮肉雏鸡器官发育的影响,探讨三者作用差异.350只1日龄肉雏鸡随机分为7组,每组5个重复,每重复10只鸡,每组分别饲喂玉米-豆粕基础日粮和外加50 mg/kg WSP、100mg/kg WSP、50 mg/kg AEP、100 mg/kg AEP、3 mg/kg木聚糖酶、6mg/kg木聚糖酶的试验日粮.试验期15日.结果表明,和对照组相比,WSP、AEP和木聚糖酶显著增加了肉雏鸡体重、法氏囊、腺胃和肌胃重量以及肠道长度(P<0.05),对直肠中乳酸菌、链球菌、沙门氏菌和大肠杆菌无显著影响.WSP和AEP增加了食糜粘度,而木聚糖酶降低了食糜粘度,二者

  4. 水分胁迫下内生真菌球毛壳ND35对冬小麦苗期生长和抗旱性的影响%Effect of the endophytic fungus Chaetomium globosum ND35 on the growth and resistance to drought of winter wheat at the seedling stage under water stress

    Institute of Scientific and Technical Information of China (English)

    丛国强; 尹成林; 何邦令; 李玲; 高克祥

    2015-01-01

    为明确不同水分条件下内生真菌对冬小麦苗期生长和抗旱性的影响,以抗旱型小麦品种山农16和水分敏感型小麦品种山农22为材料,利用荧光定量PCR技术检测小麦干旱诱导基因脱水素wzy2的表达量来了解冬小麦在干旱胁迫下相关基因的表达差异,通过测定相关生理指标与酶活性来判断小麦发育及其在干旱胁迫下的生理响应状况.结果表明,与正常水分ND35组相比,接种球毛壳菌(Chaetomium globosum)ND35的干旱处理组小麦的根冠比、总蛋白含量、脯氨酸含量及丙二醛含量等指标显著提高,小麦叶片含水量和可溶性糖含量有所降低.在干旱处理组中,球毛壳菌ND35可以显著提高小麦山农16的根长和山农22的株高,接种球毛壳ND35的山农16脯氨酸含量、可溶性糖含量、过氧化氢酶活性比对照组均显著提高,丙二醛含量比对照组降低9.0%,但差异不显著;山农22脯氨酸含量和过氧化氢酶活性比对照组显著提高,丙二醛含量和可溶性糖含量比对照组有所降低,但可溶性糖含量差异不显著;相对定量检测数据显示,接种球毛壳ND35后,两种小麦脱水素wzy2基因的表达量较对照组均能够显著提高.综合分析说明内生真菌球毛壳ND35可以促进冬小麦苗期根系和植株发育,小麦提前进入三叶期,增强小麦避旱性,同时提高小麦根系活力,增强小麦耐旱性;提高个体细胞内水分、糖分、脯氨酸含量,降低丙二醛的氧化性损伤,增强过氧化氢酶活性,从而提高两种冬小麦对干旱胁迫的耐受能力;球毛壳ND35促进小麦干旱诱导相关基因wzy2的表达量,进而提高抗旱相关蛋白的表达,从而提高两种冬小麦耐脱水性和对干旱胁迫的适应性.

  5. Research of the dietary fiber modified from apple flesh pomace using xylanase enzyme%木聚糖酶对苹果肉渣膳食纤维改性的研究

    Institute of Scientific and Technical Information of China (English)

    付成程; 郭玉蓉; 严迈; 霍天博; 薛战锋; 孙迪迪; 贾莉

    2013-01-01

    Study on the optimal process and determine physical and chemical properties of soluble dietary fiber and separated residues,with the apple flesh pomace as raw materials which were hydrolysed by xylanase.The results showed that the yield of soluble dietary fiber was 19.58% processed by xylanase.The optimal reaction conditions were as follows: the enzyme addition was 12.375U/g, reaction time was 7h, reaction temperature was 55℃,and reaction pH was 5.0.Water-soluble dietary fiber gained by xylanase had higher solubility and lower apparent viscosity than raw materials. The retention ability and expansibility of the separated residues were improved. The infrared spectrum analysis described both soluble dietary fiber and separated residues illustrated similar characteristic absorption peak as polysaccharide,accompanied by obvious changes of microstructure which could be seen under scanning electron microscope.%以苹果肉渣为原料,通过木聚糖酶酶解,确定最优工艺,并研究所得水溶性膳食纤维及分离滤渣的理化性质.结果表明:采用木聚糖酶法,得到的水溶性膳食纤维提取率为19.58%.酶解温度55℃,酶解pH7.0时其最适反应条件为:酶添加量12.375U/g、酶解时间7h.酶解所得水溶性膳食纤维有较高的溶解性,表观粘度有所降低;分离滤渣的持水力与膨胀力均提高;红外光谱分析显示,酶解所得水溶性膳食纤维以及分离滤渣均产生糖的特征吸收峰,而在电镜扫描下,超微结构变化明显.

  6. Phylogenetic analysis of the complete mitochondrial genome of Madurella mycetomatis confirms its taxonomic position within the order Sordariales.

    Directory of Open Access Journals (Sweden)

    Wendy W J van de Sande

    Full Text Available BACKGROUND: Madurella mycetomatis is the most common cause of human eumycetoma. The genus Madurella has been characterized by overall sterility on mycological media. Due to this sterility and the absence of other reliable morphological and ultrastructural characters, the taxonomic classification of Madurella has long been a challenge. Mitochondria are of monophyletic origin and mitochondrial genomes have been proven to be useful in phylogenetic analyses. RESULTS: The first complete mitochondrial DNA genome of a mycetoma-causative agent was sequenced using 454 sequencing. The mitochondrial genome of M. mycetomatis is a circular DNA molecule with a size of 45,590 bp, encoding for the small and the large subunit rRNAs, 27 tRNAs, 11 genes encoding subunits of respiratory chain complexes, 2 ATP synthase subunits, 5 hypothetical proteins, 6 intronic proteins including the ribosomal protein rps3. In phylogenetic analyses using amino acid sequences of the proteins involved in respiratory chain complexes and the 2 ATP synthases it appeared that M. mycetomatis clustered together with members of the order Sordariales and that it was most closely related to Chaetomium thermophilum. Analyses of the gene order showed that within the order Sordariales a similar gene order is found. Furthermore also the tRNA order seemed mostly conserved. CONCLUSION: Phylogenetic analyses of fungal mitochondrial genomes confirmed that M. mycetomatis belongs to the order of Sordariales and that it was most closely related to Chaetomium thermophilum, with which it also shared a comparable gene and tRNA order.

  7. 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.

  8. 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

    cell attachment. To confirm this theory, R. marinus was grown, and activity assays showed that the major part of the xylanase activity was connected to whole cells. Moreover, immunocytochemical detection using a Xyn10A-specific antibody proved presence of Xyn10A on the R. marinus cell surface......-termini of proteins that were predominantly extra-cellular/cell attached. A primary structure motif of three conserved regions including structurally important glycines and a proline was also identified suggesting a conserved 3D fold. This bioinformatic evidence suggested a possible role of this domain in mediating....... In 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 phylum...

  9. 金针菇菌糠中木聚糖酶的分离纯化及其酶学性质研究%Purification and Properties of Xylanase from Spent Mushroom Compost of Flammulina velutipes

    Institute of Scientific and Technical Information of China (English)

    杨云龙; 黄彩梅; 白植成; 胡开辉

    2015-01-01

    废弃食用菌栽培料( SMC)中含有一定活性的木聚糖酶,作者以金针菇菌糠为材料,采用盐析、Sephadex G-100凝胶层析和DEAE C-52离子交换层析等技术,对SMC中的木聚糖酶进行分离和纯化,并进行酶学性质研究。结果表明,该酶经SDS-PAGE电泳鉴定为电泳纯,分子量为37.8 KDa,酶比活力达到946.67 U/mg,纯化倍数为12.74;酶的最适温度和pH分别为50℃和pH 6.0,Fe2+、Zn2+对木聚糖酶有激活作用,Ca2+、Mn2+、K+有促进作用,Mg2+、Cu2+、Fe3+有抑制作用,反应动力学参数Vmax和Km 分别为15.43μg/mL/min、9.61 mg/mL。%Xylanase produced by Spent Mushroom Compost was isolated and purified by ammonium sulfate salting-out,Sephadex G⁃100 column chromatography and DEAE C⁃52 anion chromatography exchange,and its enzymatic properties were studied.The results illustrated that it exhibited a single band by the SDS⁃PAGE,and the molecular weight was estimated as 37. 8 KDa. The purification made the specific activity as high as 946.67 U/mg and purification multiple reached 12.74.The optimal temperature and pH value for activity were 50 ℃ and pH 6.0 respectively,which had a certain acid and alkali resistance and high⁃temperature⁃resisting.Xy⁃lanase obtained in this study was activated by Fe2+and Zn2+;promoted by Mn2+,K+and Ca2+;while inhibited by Mg2+,Cu2+ and Fe3+.The Vmax and Km values of the xylanase for xylan were 15.43μg/mL/min and 9.61 mg/mL, respectively.The study suggests that this xylanase has a good potential to be a cold⁃stable xylanase in paper man⁃ufacturing and feed industries.

  10. 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

  11. Bio-conventional bleaching of kadam kraft-AQ pulp by thermo-alkali-tolerant xylanases from two strains of Coprinellus disseminatus for extenuating adsorbable organic halides and improving strength with optical properties and energy conservation.

    Science.gov (United States)

    Lal, Mohan; Dutt, Dharm; Tyagi, C H

    2012-04-01

    Two novel thermo-alkali-tolerant crude xylanases namely MLK-01 (enzyme-A) and MLK-07 (enzyme-B) from Coprinellus disseminatus mitigated kappa numbers of Anthocephalus cadamba kraft-AQ pulps by 32.5 and 34.38%, improved brightness by 1.5 and 1.6% and viscosity by 5.75 and 6.47% after (A)XE(1) and (B)XE(1)-stages, respectively. The release of reducing sugars and chromophores was the highest during prebleaching of A. cadamba kraft-AQ pulp at enzyme doses of 5 and 10 IU/g, reaction times 90 and 120 min, reaction temperatures 75 and 65°C and consistency 10% for MLK-01 and MLK-07, respectively. MLK-07 was more efficient than MLK01 in terms of producing pulp brightness, improving mechanical strength properties and reducing pollution load. MLK-01 and MLK-07 reduced AOX by 19.51 and 42.77%, respectively at 4% chlorine demands with an increase in COD and colour due to removal of lignin carbohydrates complexes. A. cadamba kraft-AQ pulps treated with xylanases from MLK-01 to MLK-07 and followed by CEHH bleaching at half chlorine demand (2%) showed a drastic reduction in brightness with slight improvement in mechanical strength properties compared to pulp bleached at 4% chlorine demand. MLK-01 reduced AOX, COD and colour by 43.83, 39.03 and 27.71% and MLK-07 by 38.34, 40.48 and 30.77%, respectively at half chlorine demand compared to full chlorine demand (4%). pH variation during prebleaching of A. cadamba kraft-AQ pulps with strains MLK-01 and MLK-07 followed by CEHH bleaching sequences showed a decrease in pulp brightness, AOX, COD and colour with an increase in mechanical strength properties, pulp viscosity and PFI revolutions to get a beating level of 35 ± 1 °SR at full chlorine demand.

  12. 脉孢霉固体发酵产木聚糖酶的条件研究%The solid state fermentation conditions of xylanase production by Neurospora sitophila

    Institute of Scientific and Technical Information of China (English)

    宫晓; 郑喜群; 刘晓兰; 邓永平

    2015-01-01

    The solid state fermentation conditions of xylanase production by Neurospora sitophila was studied ,in order to improve the yield .Through single factors and Plackett‐Burman design method ,the optimum culture conditions and inorganic salt types were selected .The results showed that the suitable culture medium was composed of soybean powder and corn bran with the ratio of 4∶1 ;inorganic salt was copper sulfate and calcium chloride with concentration of 0 .3% and 0 .6% ,respec‐tively ;fermentation time was 60 h and extraction time was 2 h . Under the conditions ,the activity of xylanase reached 577 U/m l .%研究了好食脉孢菌固体发酵生产木聚糖酶的条件,以提高其产量。通过单因素及Plackett‐Burman设计法,初步选出生产木聚糖酶适合的培养条件和无机盐种类。结果表明:培养基主要物料是豆饼粉、玉米皮,比例为4∶1,无机盐为0.3%硫酸铜与0.6%氯化钙,发酵时间为60h,浸提时间为4h,在此优化条件下,木聚糖酶活力可达577 U/m l。

  13. 中度嗜盐菌产木聚糖酶发酵条件的研究%Study on Fermentation Conditions of A Moderately Halophilic Bacterium Producing Xylanase

    Institute of Scientific and Technical Information of China (English)

    韩秋菊; 高飞

    2011-01-01

    中度嗜盐菌在盐碱环境下生长繁殖,其产生的木聚糖酶也同样具有在盐碱环境下发挥作用的特性.本文对一株中度嗜盐菌的产木聚糖酶活性进行了初步研究.研究包括氮源、液体种子接种量、培养温度、pH值、培养时间等因素对该菌株产木聚糖酶能力的影响.结果表明,最佳培养氮源为蛋白胨;最佳产生木聚糖酶的发酵条件是液体种子接种量为6%,温度为35℃,pH值7,培养时间为4 d.%Moderately halophilic bacteria can grow and propagate under high saline environment, the producing xylanase also has the characteristics of playing roles in the high salinity and alkalinity environment. We studied its activity of xylanase, the effects of nitrogen sources, inoculating quantity of liquid seed, temperature, pH value and culture time. The results showed that the optimal nitrogen source was peptone and the optimum conditions produ cing the enzyme were as follows: the liquid inoculation of 6%, culture temperature of 35℃, initial pH of 7 and culture time of 4d.

  14. 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.

  15. Purification and Properties of Xylanases from Pencillium corylophilum%顶青霉木聚糖酶的纯化与性质

    Institute of Scientific and Technical Information of China (English)

    杨瑞金; 许时婴; 王璋

    2001-01-01

    Three parts of xylanases (Part A, Part B and Part C) wereseparated and purified from a culture filtrate of Pencillium corylophilum No.P-3-31. Part B was further purified to homogeneity and Part C was further purified to almost homogeneity by the same procedures as Part B. The molecular weights of Part B and Part C were estimated to be 24 200 and 48 300 respectively by SDS-PAGE. The optimal pH and temperature of enzymes were 4.0 and 45 ℃ for Part A and Part B, while 5.5 and 50~55 ℃ for Part C. Part C showed a substrate-cross- specificity on hydrolyzing CMC and had an activity of β-xylosidase, but this activity was unable to hydrolyze xylobiose. Part A and Part B did not show the substrate-cross-specificity. Both crude and purified enzymes showed significant differences in their activities to hydrolyze different xylans from different sources. For the crude enzyme, if the activity to birchwood xylan was set as 100%, then those to corncob xylan and bagasse xylan were 143% and 124% respectively.%从顶青霉(Pencilliumcorylophilum)P-3-31培养液中分离到3种木聚糖酶组分,分别称为PartA、PartB和PartC.PartB进一步纯化,经SDS-PAGE鉴定为单带,相对分子质量为24200;PartC进一步纯化,经SDS-PAGE鉴定也是单带,相对分子质量为48300.PartA和PartB的最适反应条件为pH4.0,45℃;PartC的最适反应条件为pH5.5,55℃.PartA和PartB对木聚糖以外的底物不能水解;PartC具有水解CMC的交叉活性和β-木糖苷酶活性,但不能将木二糖水解成木糖.3个纯化酶组分和粗酶对不同来源的木聚糖底物均表现出不同的活性.对于粗酶,若桦木木聚糖为底物的相对酶活为100%,则玉米芯木聚糖为底物的相对酶活为143%,蔗渣木聚糖为底物的相对酶活为124%.

  16. 黄瓜内生Ch1001菌株种子处理防治根结线虫的施用剂量研究%Optimization of Seed Treatment Dose of the Cucumber Endophytic Chaetomium Ch1001 Strain for Biological Control of Root Knot Nematode Meloidogyne incognita

    Institute of Scientific and Technical Information of China (English)

    鄢小宁; 杨艳; 贺春萍; 郑经武

    2011-01-01

    The cucumber endophytic Chaetomium strain Ch1001 has shown its ability to suppress root knot nematode, Meloidogyne incognita, when applied as seed treatment. However, its optimum application dose, which is an important parameter for practical use in order to obtain robust seedlings as well satisfied nematode control efficacy has not been determined. In this paper, the root viability, defense related enzymes activity as well root knot nematode control efficacy were tested after cucumber seeds were inoculated with Ch1001 at 1×103, 1×104, 1×105,1×106, 1×107, 1×108ascospores/seed respectively at sowing. Results indicated that root viability was not influenced by Ch1001 when the inoculum dose was at or lower than 1×105 ascospores/seed,but increased significantly at the inoculum dose of 1×106 and 1×107 ascospores/seed, and decreased dramatically at 1×108 ascospores/seed. PAL and chitinase activity did not enhance after Ch1001 treatment, POD and CAT activity significantly increased at 1×105, 1×10' and 1×107 ascospores/seed, indicating that Ch1001 induced resistance related to active oxygen metabolism. The root knot nematode M. incognita control efficacy was positively correlated to seed treatment dose, the galls formation was significantly inhibited at 1×104,1×105,1×106 and 1×107 ascospores/seed treatment, the satisfied inhibition reached at 1×106 and 1×107 ascopores/seed treatment, which were 45.3% and 57.0% respectively. Based on the above results, the dose at 1 ×107 ascospores/seed was optimum for practical use because of the enhanced root viability, defense related enzymes activity as well as the lowest nematode invasion.%内生毛壳菌Ch1001菌株种子处理对黄瓜幼苗上发生的根结线虫有防效.为培育壮苗并达到理想的防效,合适的施用剂量是实际应用所需的一个重要参数.笔者在黄瓜播种时分别用1×103、1×104、1 ×105、1× 106、1×107、1×108个孢子/种子的剂量处理种子,测定幼苗的

  17. Utilization of feruloyl esterase and xylanase for the degradation of brewers' spent grain%阿魏酸酯酶和木聚糖酶协同降解麦糟

    Institute of Scientific and Technical Information of China (English)

    李夏兰; 程珊影; 杨道秀; 方柏山

    2012-01-01

    橘青霉以麦糟为唯一碳源培养时,阿魏酸酯酶的最佳发酵时间为60 h,其酶活力可达40.8 mU/mL。在pH值为5.0、45℃、料液比1∶30(g∶mL)条件下,取47.5 U/mL的木聚糖酶粗酶液15 mL,加入1.0 g麦糟的乙醇不溶物,反应12 h后,加入40.8 U/mL的阿魏酸酯酶粗酶液15 mL再反应12 h,阿魏酸和低聚木糖释放率分别为54.1%和161 mg/g(麦糟的乙醇不溶物)。实验结果还表明,阿魏酸酯酶与木聚糖酶存在协同作用,能极大提高麦糟中阿魏酸及低聚木糖的释放率,有利于麦糟的降解。%The optimal fermentation time ofPenicillium citrinum feruloyl esterase ( PcFAE ) was 60 h and its activity was 40.8 mU/mL when the Brewers' Spent Grain ( BSG ) was used as the sole carbon source for Penicillium citrinum. One gram of alcohol-insoluble residue ( AIR ) of Brewers'spent grain ( BSG ) was processed by the solution of crude xylanase ( XG180 ) ( 47.5 U/mL, 15 mL ) for 12 hours at the condition of the material-liquid ratio 1 : 30( W/V ), pH 5.0 and 45 ℃, respectively. And then the solution of crude PcFAE ( 40.8 mU/mL )was added and the sample was processed for another 12 h. The maximum release rate of Ferulic acid ( FA ) was 54.1% and the release amount of xylooligosaccharide (XOS) was 161 mg/g (BSG-AIR), respectively. The results of our experiments showed that the release rate of FA and XOS from BSG-AIR increased significantly as the PcFAE could coordinate with the xylanase and enzymolysis process was also conducive to the degradation of BSG.

  18. 产耐热木聚糖酶细菌的分离鉴定及酶易错PCR致突变条件优化%Isolation & Identification of a Heat-Resistant Xylanase-Producing Bacterial Strain & Optimization of the Enzyme Error-Prone PCR Mutagenic Conditions

    Institute of Scientific and Technical Information of China (English)

    赵超; 张宁宁; 梅凡; 艾超; 阮灵伟; 黄一帆; 刘斌

    2013-01-01

    从福建省永泰县温泉采集样品中筛选到1株产耐热木聚糖酶嗜热菌株TC-W7,并获得该木聚糖酶基因。在此基础上,采用易错PCR技术在木聚糖酶基因中引入突变,研究Mg2+浓度、Mn2+浓度、dTTP/dCTP浓度等条件对突变率的影响。通过形态特征、生理生化试验及16S rRNA序列相似性比对分析,初步鉴定菌株TC-W7为土壤芽胞杆菌(Geobacillus),菌株TC-W7在最适温度75℃和 pH 8.2条件下,其木聚糖酶活力为215.83 U/mL,Triton X-100和DDT能显著增强该酶的活性。在 Mg2+浓度为20μmol/L,Mn2+浓度为0.80μmol/L,dTTP/dCTP浓度为0.30 mmol/L的致突变条件下,碱基突变率为0.98%。 Geobacillus sp. TC-W7产木聚糖酶具有较好的耐热和耐碱等工业应用特性,对该酶易错PCR致突变条件优化结果,可用于后续木聚糖酶的耐热定向进化。%A heat-resistant xylanase-producing bacterial strain TC-W7 from samples collected in a hot spring in Yong-tai County, Fujian Province was screened and obtained xylanase gene of the strain. Based on these an error-prone PCR ( Ep-PCR) technique was adopted to introduce mutation in the xylanase gene, to study the effects of the concentration such as Mg2+, Mn2+ and dTTP/dCTP and other conditions on the mutation rate. It was initially identified that strain TC-W7 belonged to Geobacillus through morphology features, physiological and biochemical tests as well as 16S rRNA sequence comparative analysis. Under the most suitable temperature 75℃ and pH 8. 2, the activity of xylanase was at 215. 83 U/mL, Triton X-100 and DDT could remarkably increase the activity of xylanase. The base mutation rate was at 0. 98% under the mutagenic conditions of 20. 0 μmol/L Mg2+, 0. 80 μmol/L Mn2+ and 0. 30 mmol/L dTTP/dCTP. The xylanase-producing Geobacillus sp. TC-W7 had a fine heat and alkali resistance and other industry appli-cable features. The results of Ep-PCR mutagenic conditions optimization of the enzyme can be used for

  19. Xylanase production by Trichoderma harzianum rifai by solid state fermentation on sugarcane bagasse Produção de xilanase pelo Trichoderma harzianum rifai por fermentação em fase sólida em bagaço de cana de açúcar

    Directory of Open Access Journals (Sweden)

    Maria Inês Rezende

    2002-01-01

    Full Text Available Sugarcane bagasse was used as substrate for xylanase production by means of a strain of Trichoderma harzianum Rifai isolated from decaying Aspidosperma sp. (peroba wood. The bagasse was washed, dried, milled and wetted with minimal salts medium and the cultures grown at 28 ± 2ºC for 7 days. Two extraction methods were tested for enzyme recovery: (A Tween 80, 0.1% (v/v, in physiological saline, and (B 50mM sodium acetate buffer, pH 5.0, under agitation (180rpm for 15, 30 and 60min. After a single extraction, both extraction methods recovered an average of 15U/ml of xylanase activity, independent on the time of shaking. A second and third extraction recovered 10.4 and 6.6U/ml xylanase, respectively. The effect of volume size for extraction, and sugarcane bagasse concentration, on xylanase production were also investigated. The growth profile of Trichoderma harzianum was followed over 20 days on 14% (w/v bagasse, and highest xylanase activity (288U/ml appeared on the seventh day. The enzymatic extract after precipitation with ammonium sulphate was submitted to electrophoresis on polyacrylamide gels and showed 4 protein-staining bands, one of which exhibited xylanase activity.Bagaço de cana-de-açúcar foi utilizado como substrato para a produção de xilanase pelo Trichoderma harzianum Rifai, isolado previamente de peroba (Aspidosperma sp em decomposição. Após ter sido lavado, seco e moído, o bagaço foi umedecido com meio mínimo de Vogel e autoclavado. Dois métodos de extração foram avaliados para recuperação da enzima: (A Tween 80 0,1% em salina fisiológica (v/v e (B tampão acetato pH 5 (50mM sob agitação (180rpm durante 15, 30 e 60 minutos. Em média, foram recuperados 15U/ml de atividade xilanásica com ambos os extratores, após uma única extração, independente do tempo de agitação. Uma segunda e terceira reextrações recuperaram 10,4 e 6,6U/ml, respectivamente. O efeito de diferentes volumes de extração e a concentra

  20. Contribution of protein, starch, and fat to the apparent ileal digestible energy of corn- and wheat-based broiler diets in response to exogenous xylanase and amylase without or with protease.

    Science.gov (United States)

    Romero, L F; Sands, J S; Indrakumar, S E; Plumstead, P W; Dalsgaard, S; Ravindran, V

    2014-10-01

    The ileal energy contribution of protein, starch, and fat in response to 2 exogenous enzyme combinations was studied in 2 digestibility assays with 21- (experiment 1; 432 birds) and 42-d-old (experiment 2; 288 birds) Ross 308 broiler chickens. A 2 × 2 × 3 factorial arrangement of treatments with 2 base grains (corn or wheat), without or with high fiber ingredients (corn distillers dried grains with solubles and canola meal), and 3 enzyme treatments was implemented. Enzyme treatments, fed from 12 to 21 d or 32 to 42 d, were 1) without enzymes, 2) with xylanase from Trichoderma ressei (2,000 U/kg) and amylase from Bacillus licheniformis (200 U/kg; XA), or 3) with XA plus protease from Bacillus subtilis (4,000 U/kg; XAP). All diets contained Escherichia coli phytase (500 FTU/kg). Apparent ileal digestibility (AID) of protein, starch, and fat, as well as the apparent ileal digestible energy, were determined using titanium dioxide as inert marker. A generalized mixed model was used to test main effects and 2-way interactions at P starch at 21 and 42 d, and AID of fat at 21 d, with greater effects of enzymes in wheat-based compared with corn-based diets, but significant increments due to enzymes compared with controls in both diet types. Apparent ileal digestibility of fat at 42 d increased with enzyme supplementation compared with the control treatments. The XA and XAP treatments gradually (P starch, fat, and protein were affected differentially by base grain and the presence of fibrous ingredients at 21 and 42 d of age.

  1. The Effect of Triticale and Enzyme Cocktail (Xylanase & β-Glucanase Replacement in Grower Diet on Performance, Digestive Organ Relative Weight, Gut Viscosity and Gut Morphology of Broiler Chickens

    Directory of Open Access Journals (Sweden)

    Heydar Zarghi

    2016-11-01

    Full Text Available Introduction Corn and wheat are the grains most routinely used in commercial poultry diets. For consumption of these cereals, there are a competition between humans and mono-gastric animals. Due to corn crop limitation in Iran, approximately 50% of the corn required for poultry nutrition is supplied through imports. Since triticale is more resistant to various diseases, dry weather and in similar culture and weather conditions can produce higher yield than wheat, triticale considered as a crop suitable for cultivation in inefficient lands and its culture in the world has being increased. The use of triticale in broiler feed has been limited because of the presence of soluble non-starch polysaccharides components. The purpose of this study was to evaluate the effects of different replacement levels of corn by triticale in grower diets with and without exogenous enzyme supplementation on growth performance, relative weight of digestive organs, jejunal morphology, and intestinal viscosity of broiler chickens. Materials and Methods Five hundred 11 d old male broiler chicks (Ross 308, were assigned to a factorial arrangement (5×2 with a completely randomized design with 5 replicates of 10 chicks each. The factors included 5 levels of triticale replacement levels for corn (0, 10, 20, 30 and 40% and 2 levels (zero and 0.5 g /kg of diet of enzyme cocktail “Xylanase & β-Glucanase” in the broiler grower diets. The experimental diets were isocalric and isonitrogenous and fed ad-libitum from 11 to 24 d of age. The growth performance as mean body weight at 24 d of age, daily weight gain, daily feed intake, and feed conversion ratio were calculated. At 24d of age, one bird from each pen, close to the average pen weight was selected, weighed, and euthanized by cervical dislocation. The gastrointestinal (GI tract organs were emptied and weighed. Approximately 1.5 g of wet weight of the fresh digesta was immediately placed in a micro centrifuge tube and

  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

    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

  3. 厌氧菌群SVY42产酶条件分析及产木聚糖酶菌株的分离鉴定%Enzyme-Producing Conditions Analysis of Anaerobic Bactearial Population SVY42 andIsolation and Identification of a Xylanase-Producing Strain

    Institute of Scientific and Technical Information of China (English)

    赵超; 李婷; 邓云金; 刘晓艳; 黄一帆; 刘斌

    2013-01-01

    A highly effective and stable cellulose and hemicellulose degradable anaerobic bacterial population (ABP) SVY42 from the samples collected from Great Basin Hot Spring in Nevada,USA as material was enriched with and obtained.Their production conditions of CMCase,β-galactosidase and xylanase by ABP SVY42 were studied with the giant Juncao,bagasse,waste mushroom culturing cylinder,sodium carboxymethyl cellulose (CMC),filter paper and xylan as carbon sources.Based on these,a xylanase high-producing strain was screened using xylan as substrate.The highest β-galactosidase activity was 0.23 U/mL using the giant Juncao as substrate.The highest CMCase and xylanase activities were 0.31 U/mL and 0.35 U/mL using xylan as substrate respectively.A xylanase high-producing thermophilic strain SVY42-1 from ABP SVY42 was screened from ABP SVY42.The xylanase activity reached 0.26 U/mL under the optimal temperature (41℃) and pH (8.0).Strain SVY42-1 was identified by 16S rDNA sequencing analysis and only 93.8% similar to the highest homology of the known strains,and identified initially belong to a new genus.%以美国内华达州大盆地温泉采集样品为材料,富集获得纤维素及半纤维素高效稳定降解厌氧菌群SVY42,以巨菌草、甘蔗渣、废菇筒、羧甲基纤维素钠、滤纸、木聚糖为碳源,分析菌群SVY42产内切葡聚糖酶(CMC酶)、β-葡萄糖苷酶和木聚糖酶的情况.在此基础上,以木聚糖为底物筛选高产木聚糖酶的菌株.菌群SVY42在以巨菌草作为碳源时的β-葡萄糖苷酶活最高为0.23 U/mL,以木聚糖作为碳源时CMC酶活和木聚糖酶活均为最高,分别为0.31 U/mL和0.35 U/mL.从菌群SVY42中筛选得到1株高产木聚糖酶厌氧菌株SVY42-1,该菌在最适温度41℃和pH 8.0条件下,其木聚糖酶活力为0.26 U/mL,对其进行16S rDNA序列系统进化分析,SVY42-1与已知菌株的最高同源性仅为93.81%,初步鉴定属于新属.

  4. 一株产木聚糖酶放线菌的液体发酵条件优化及水解特性研究%Optimization of Liquid Fermentation Conditions for Xylanase Production by a Actinomyces and Characterization of the Enzyme

    Institute of Scientific and Technical Information of China (English)

    朱运平; 禇文丹; 李秀婷; 滕超; 李娥; 杨然

    2012-01-01

    以木聚糖为唯一碳源制作选择培养基,利用透明圈法筛选高产木聚糖酶菌株,对其中一株产酶较高的菌株L10608进行液体发酵条件优化并对所产木聚糖酶的水解特性进行研究。结果表明:菌株L10608最佳产酶条件为以质量浓度25g/L、80目的水不溶性玉米芯木聚糖为碳源,10g/L大豆蛋白胨和5g/L酵母浸膏为复合氮源,初始pH6.0、培养温度40℃、转速200r/min、表面活性剂吐温-80质量浓度4g/L,最佳产酶条件下木聚糖酶活力达1074.8U/mL。以桦木木聚糖、榉木木聚糖和燕麦木聚糖为底物研究菌株L10608所产木聚糖酶的水解特性,结果表明该木聚糖酶为内切型木聚糖酶,水解主要产物为木二糖和木三糖。表明菌株L10608有望作为功能性低聚木糖的生产菌株。%In the current study, high-xylanase-producing strain was screened from different soil samples by transparent circle method, using xylan as the only carbon source in medium. The cultural condition for xylanase production by strain L10608 was optimized and the hydrolysis property of the enzyme was further investigated. The results indicated that the optimum fermentation medium contained a carbon source of water-insoluble xylan (80 mesh) of 25 g/L, compound nitrogen source of soya peptone of 25 g/L and yeast extract of 5 g/L, initial pH 6.0, cultural temperature 40 ℃, rotational speed of 200 r/min, surfactant polysorbate 80 of 4 g/L. Under the optimized condition, the enzyme activity reached 1074.8 U/mL. The xylanase utilized all birch wood xylan, beech wood xylan and oat xylan as the substrate, exhibiting that xylanase produced by L10608 was endo-xylanase with xylobiose and xylotriose as the major hydrates. These results showed that strain L10608 could hopefully be used for industrial production of functional xyloolygosaccharides.

  5. 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.

  6. Effect of Inducers on Xylanase Activity and Biosynthesis of Ferulic Oligosaccharides in Aureobasidium pullulans%诱导物对出芽短梗霉木聚糖酶活力和阿魏酰低聚糖合成的调控影响

    Institute of Scientific and Technical Information of China (English)

    张雨青; 余晓红; 顾振新; 屠康

    2012-01-01

    Feruloyl oligosaccharides(FOs) could be produced as a result of the hydrolysis of wheat bran fiber by xylanase synthesized during fermentation by a mutant strain of Aureobasidium pullulans without the secretion of melanin bred using wheat bran.The effects of inducers such as carbon source,nitrogen source,metal ion and surfactant on the activity of xylanase and the formation of FOs were investigated.The medium composition for FOs production by the strain was optimized using the orthogonal array design method.The highest Fos production and xylanase activity,627 nmol/L and 52.66 IU/mL,respectively,were achieved by using a medium comprised of 50 g/L wheat bran,15 g/L glucose,1 g/L peptone and 1 g/L corn syrup.%以麦麸为原料诱变选育的不产黑色素出芽短梗霉为发酵菌株,利用其发酵过程中合成的木聚糖酶水解麦麸纤维,制备阿魏酰低聚糖(FOs)。研究碳源、氮源、金属离子和表面活性剂等诱导物对出芽短梗霉木聚糖酶活力和FOs合成的影响,以探明各物质对出芽短梗霉木聚糖酶活力和FOs合成的影响;利用正交试验设计方法研究各物质添加量对出芽短梗霉产酶和FOs的影响,确定芽短梗霉发酵制备FOs的最佳培养基。结果表明:50g/L麦麸处理液中添加15g/L葡萄糖、1g/L蛋白胨和1g/L玉米浆,FOs产量和木聚糖酶活力最高分别达到627nmol/L和52.66IU/mL。

  7. 嗜热脂肪土芽孢杆菌木聚糖酶基因的合成及其在大肠杆菌中的表达%De novo Synthesis and Expression of a Thermostable Xylanase from Geobacillus stearothermophilus in Escherichia coil

    Institute of Scientific and Technical Information of China (English)

    张志刚; 裴小琼; 吴中柳

    2009-01-01

    The endoxylanase XT6 secreted from Geobacillus stearothermophilus is a particularly attractive candidate for some industrial purposes and was used successfully on an industrial-scale mill trial. The gene was de novo synthesized with the codons adjusted to fit the bias of that of Escherichia coli and constructed into vector pET28a (+). After optimizing the expression conditions, functional xylanase XT6 was over expressed in E. coll with up to 65% of total protein. A maximum xylanase activity of 3,030 U/mL was obtained from cell extract against birchwood xylan. The recombinant XT6 was partly characterized and was similar with those of the native enzyme in G. stearothermophilus. This is the first report on the over expression of a de novo synthesized xylanase XT6 gene from Geobacillus stearothermophilus. Fig 6, Tab 1, Ref 19%来自嗜热脂肪土芽孢杆菌的木聚糖内切酶XT6在工业上有着重要的应用,已经成功应用于工业规模的生产试验.本文作者在合成XT6基因全序列的同时对其密码子进行了优化,且构建重组质粒在大肠杆菌中高表达.通过优化表达条件,功能正常的XT6基因在大肠杆菌中成功过量表达,蛋白表达量占细胞中总蛋白的65%.重组表达的木聚糖内切酶XT6特性和天然酶相似,以桦木木聚糖为底物测定细胞提取物中木聚糖酶活性,最大活性高达3 030 U/mL.本文首次报道了来自嗜热脂肪土芽孢杆菌中木聚糖酶基因全序列的合成和在大肠杆菌中成功过量表达.图6表1参19

  8. Implantation phaeohyphomycosis caused by a non-sporulating Chaetomium species. Phaeohyphomycose d’inoculation causée par une espèce non sporulante de Chaetomium

    NARCIS (Netherlands)

    Najafzadeh, M.J.; Fata, A.; Naseri, A.; Saradeghi Keisarid, M.; Farahyarf, S.; Gangbakhsh, M.; Ziaee, M.; Dolatabadi, S.; de Hoog, G.S.

    2013-01-01

    We report the case of a 66-year-old Iranian woman with a phaeohyphomycotic cyst (approximately 3 × 2.5 cm 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 examin

  9. 木聚糖酶诱导释放负电荷及其对漂白硫酸盐浆纤维胶体作用和留着的影响%Xylanase-induced liberation of negatively charged species and their effect on colloidal interactions and the retention of bleached kraft pulp ifbers

    Institute of Scientific and Technical Information of China (English)

    苗成; 刘忠

    2016-01-01

    The ability and specificity of various monocomponent endo-1,4-β-xylanases to release negatively charged species from never-dried, bleached, birch kraft pulp was studied. The effects of dissolution of these xylan-based components on pulp ifltrate properties and the subsmoluent chemical retention were determined. The results revealed that the amount of charged species released depended on the xylanase and that the ratio of charged species released to dissolved xylan is not linear. Chemical retention tests showed that high levels of dissolved xylan interfere with the ifxation of colloidal species, which was conifrmed by removing the dissolved hemicelluloses. The roles of residual hemicellulose and the properties of modified fibers on chemical retention and the level of internal sizing are discussed.%本文研究未干燥的漂白硫酸盐浆中各种内切-1,4-β-木聚糖酶组分释放负电荷的能力和特异性,同时分析木聚糖组分溶解液对浆料滤液性能和化学留着的影响。实验结果表明,电荷释放的数量和木聚糖酶相关,溶解木聚糖的质量和电荷释放量不成线性关系。化学留着实验表明,高含量溶解木聚糖能够影响胶体的固化作用,因为该过程能够移除溶解的半纤维素。本文探究残留的半纤维素和改性纤维性能对化学留着的作用和浆内施胶的影响。

  10. Dynamics and Modes of Action of Xylanases fromPenicillium corylophilum on Xylan%顶青霉木聚糖酶的水解动力学与水解模式

    Institute of Scientific and Technical Information of China (English)

    杨瑞金; 许时婴; 王璋

    2001-01-01

    从顶青霉(Penicillium corylophilum)培养液中分离纯化得到的3种木聚糖酶组分(Part A, Part B 和 Part C),对其水解动力学和水解模式的研究结果表明,3种木聚糖酶的动力学常数(桦木木聚糖为底物)分别为:Part A,Km=1.00 mg/mL,Vmax=0.159 U/mL;Part B, Km=1.59 mg/mL, Vmax=0.274 U/mL;Part C,Km=0.85 mg/mL,Vmax=0.200 U/mL. 3种木聚糖酶的水解模式相同,主要从一端(非还原端)水解得到木三糖和木二糖,并且得到木三糖的速度大于得到木二糖的速度;水解会产生木四糖,但木四糖会很快进一步水解成木二糖,木四糖水解成木糖和木三糖的可能性极小;木三糖会进一步水解成木糖和木二糖,但速度不快;木二糖不会进一步水解。粗酶水解玉米芯木聚糖时,阿拉伯糖侧链的水解与木聚糖主链的水解同步进行,粗酶中含有水解阿拉伯糖基侧链的阿拉伯糖苷酶活力.%Dynamics and mode of acting on xylan of three parts of xylanases (Part A, Part B and Part C) separated and purified from a culture filtrate of Penicillium corylophilum No. P-3-31 were investigated. The Km and Vmax of the three purified enzymes with birchwood xylan as a substrate were 1.00 mg/mL and 0.159 U/mL for Part A ( pH 4.0, 50 ℃) , 1.59 mg/mL and 0.274 U/mL for Part B (pH 4.0,50 ℃) and 0.85 mg/mL and 0.200 U/mL for Part C (pH 5.5,50 ℃). Based on the dynamic studies, it was deduced that three purified enzymes had the same mode of action on xylan. Xylotriose and xylobiose were main hydrolysates of long-chain xylan by these enzymes. Xylooligosaccharides upwards from xylotetraose were immediately hydrolyzed, and xylotetraose was mainly hydrolyzed to xylobiose. Xylotriose was slowly hydrolyzed, but xylobiose was unable to be further hydrolyzed by these enzymes. The studies on the hydrolysis dynamics of xylan from corncob with crude enzyme showed that the hydrolysis of arabinose side-chain was almost

  11. 不同木聚糖水平饲粮中添加木聚糖酶对断奶仔猪生长性能及肠道微生态环境的影响%Xylanase Supplementation of Diets Containing Different Levels of Xylan for Weaned Piglets: Effects on Growth Performance and Intestinal Microecology

    Institute of Scientific and Technical Information of China (English)

    叶楠; 陈代文; 毛湘冰; 黄志清; 毛倩; 陈洪; 韩国全; 余冰

    2011-01-01

    A 2 x 3 factorial design was conducted to study the effects of dietary xylan level and xylanase supplementation on growth performance, intestinal microbial metabolites and intestinal flora of weaned piglets. Tow main factors were xylanase supplementation (0 and 1 000 U/kg) and dietary xylan level (4. 58% , 6.23% and 7. 54% ). Thirty cross-bred (Duroc x Landrace x Large white) weaned piglets at 28 days of age with the similar weight and genetic background were randomly allotted to 6 treatments with 5 replicates each and 1 piglet in each replicate. The experiment lasted for 14 days. The results showed that during the experimental period from 1 to 14 days, the supplementation of xylanase increased the average daily gain (P 0. 05), and improved the total volatile fatty acid content of ileum digesta (P <0. 05), reduced the ammonia content of caecum digeta (P <0. 05), and increased the amount of Bifidobacterium in caecum digeta (P <0.05). When dietary xylan level was 7. 54% , the most significant effect of xylanase supplementation was found in those indexes ( P <0.05). However, dietary xylan level did not significantly affect those indexes. In conclusion, under the condition of this trial, the supplementation of xylanase can improve growth performance and intestinal microenvironment of weaned piglets. In addition, there is a dose-dependent relationship between xylanase supplementation and dietary xylan level.%本试验旨在研究不同木聚糖水平饲粮中添加木聚糖酶对断奶仔猪生长性能、肠道微生物代谢产物和菌群数量的影响.选择30头体重相近,遗传背景相似的28日龄“杜×长×大”三元杂交断奶阉公猪,采用2×3因子试验设计,设2个饲粮木聚糖酶添加量(0和1 000 U/kg)和3个饲粮木聚糖水平(4.58%、6.23%和7.54%),将仔猪随机分为6个处理,每个处理5个重复,每个重复1头猪,试验期14 d.结果表明:试验l~14 d,添加木聚糖酶能显著提高仔猪平均日增重(P<0.05)

  12. Studies on Secreted Expression of an Acidic Xylanase in Pichia pastoris and Evaluation of its in vitro Activity%一种酸性木聚糖酶在毕赤酵母中的分泌表达及体外活性评价研究

    Institute of Scientific and Technical Information of China (English)

    杨雯涵; 郭晓晶; 陈轶群; 吕俊楠; 谢飞; 曹云鹤

    2013-01-01

    通过构建硫色曲霉酸性木聚糖酶基因xynA串联双拷贝工程菌,提高酶的发酵活力。10 L发酵罐中,双拷贝菌株发酵活力达3574 U/mL。重组木聚糖酶的最适催化温度为50℃,最适反应pH为2.2~3.4,在pH 1.7的酸性缓冲液下保持100 min,酶活残留70%以上。重组木聚糖酶对胃蛋白酶和胰蛋白酶也具有较好的耐受性。该木聚糖酶在低pH条件下的高催化活性、耐酸性及蛋白酶耐受性,使之具有较好的应用前景。通过体外模拟猪胃肠道环境以快速评价重组酶的作用效果。试验以小麦型日粮作为基础日粮,设计4个处理组,重组酶的添加量分别为0 U/kg、2000 U/kg、4000 U/kg和6000 U/kg,每个处理5个重复,使用SDS-Ⅱ单胃动物仿生消化系统模拟猪的胃肠道环境消化试验日粮,消化后的残渣烘干后测定其总能、粗蛋白质、中性洗涤纤维含量以计算其消化率。结果表明,添加重组酸性木聚糖酶显著提高了总能、粗蛋白质(线性和二次;P<0.01)和中性洗涤纤维的消化率(线性和二次;P<0.05)。重组酶的最适宜添加量为4000 U/kg。%In this study, engineering Pichia pastoris containing double copies of Aspergillus sulphureus xylanase gene xynA was constructed in order to earn a high expression level and low production cost. In a 10 L fermentor, double-copy recombinant strain yielded the enzyme activity of 3 574 U/mL. The recombinant xylanase exhibited optimal activity at 50℃ and pH 2.2~3.4. Residual activity of the raw recombinant xylanase was over 70%, when fermentation broth was pretreated with Na2 HPO4-citric acid buffer of pH 1.7 for 100 min. And the recombinant xylanase exhibited strong resistance to pepsin and trypsin. High level of catalytic activity in low pH and acid resistance suggested that the recombinant xylanase has a prospective application in feed industry as an additive. Then the recombinant enzyme

  13. Expression of a Xylanase Gene Originated from Rumen Anaerobic FungiNeocallimastix frontalis inPichia pastoris%来源于瘤胃厌氧真菌Neocallimastix frontalis木聚糖酶在毕赤酵母中的表达

    Institute of Scientific and Technical Information of China (English)

    汪艳; 李晓; 陈勇; 武运

    2015-01-01

    厌氧真菌Neocallimastix frontalis是瘤胃中降解木聚糖和纤维素的主要微生物之一,其木聚糖酶具有潜在的应用价值.对来源于Neocallimastix frontalis木聚糖酶基因Xyn11B进行密码子优化;通过全基因合成优化后的木聚糖酶基因Xyn11Bm,构建该基因的酵母表达载体pPIC9K-Xyn11Bm,并在毕赤酵母GS115中诱导表达.摇瓶水平时,重组Xyn11Bm酶活性最高为4 874.8 U/mL.在10 L发酵罐中诱导96 h后,重组Xyn11Bm的酶活性为5 139.7 U/mL,菌体湿重和干重达到216.7 g/L和117.3 g/L.酶学性质分析表明,重组Xyn11Bm的最适反应温度为50℃,最适反应pH为5.0.在pH5.0-8.0时该酶具有较好的稳定性,但温度稳定性较差.底物特异性分析表明,重组Xyn11Bm可水解燕麦木聚糖、桦木木聚糖和可溶性木聚糖4-O-Me-D-glucurono-D-xylan,但不降解地衣多糖和大麦β-葡聚糖.结果表明重组Xyn11Bm具有潜在的应用价值.%The anaerobic fungus Neocallimastix frontalisis one of main microorganisms in the rumen degrading xylan and cellulose and its xylanase has the potential application value. In this study, a xylanase gene Xyn11B originated from N. frontaliswas codon optimized, and the optimized gene Xyn11Bm was synthesized. Based on the gene engineering technology, the yeast expression vector pPIC9K-Xyn11Bm was constructed, and the xylanase was induced and expressed in Pichia pastoris GS115. In shake flask level, enzyme activity of the recombinant Xyn11Bm reached the maximum up to 4874.8 U/mL. In 10 L fermentor, at 96 h after induction, the activity of recombinant enzyme was 5139.7 U/mL, cell wet weight and dry weight were 216.7 g/L and 117.3 g/L. Enzymatic properties analysis showed that the optimum reaction temperature and pH of Xyn11Bm were 50℃ and 5.0. In pH5.0-8.0, the enzyme had sound stability, but poor temperature stability. Substrate specificity analysis showed that recombinant Xyn11Bm could hydrolyze oat spelt xylan, birch xylan and soluble

  14. BÚSQUEDA DE LAS MEJORES CONDICIONES PARA LA EXTRACCIÓN Y MEDIDA DE ACTIVIDAD DE CELULASA Y XILANASA EXTRAÍDAS DE LA CORTEZA DE PITAYA AMARILLA (Acanthocereus pitajaya Searching the Best Conditions for the Extraction and Activity Measurement of Cellulase and Xylanase Extracted from the Yellow Pitaya Fruit Peel (Acanthocereus pitajaya

    Directory of Open Access Journals (Sweden)

    YENNY MARITZA DUEÑAS GÓMEZ

    Full Text Available Para pitaya amarilla (Acanthocereus pitajaya se ha encontrado que el ablandamiento excesivo de la cáscara contribuye al deterioro del fruto, al aplicar diferentes técnicas de conservación en fresco. Dado que tanto la celulasa como la xilanasa se han vinculado con el ablandamiento de la cáscara de frutos, este trabajo se basó en la búsqueda de las mejores condiciones de extracción y medida de actividad de celulasa y xilanasa. El mejor sistema de extracción fue buffer fosfato 20 mM, NaCl 0,5 M, pH 7,0. Para la medida de actividad de celulasa es necesario incubar durante 60 min a 37 ºC, con un volumen de extracto enzimático crudo de 30 µL, empleando buffer acetato 100 mM a pH 5,0; los valores de constante aparente de Michaelis Menten (K M aparente y velocidad máxima (V MÁX fueron 0,279 mg/mL y 0,00014 nmol glucosa/min, respectivamente. Para determinar la actividad de xilanasa se establecieron 15 min de tiempo de incuba-ción, a 50 ºC, empleando 30 µL de extracto enzimático crudo a pH 4,0 (buffer acetato 100 mM; los valores de K M aparente y V MÁX para xilanasa fueron 0,073 mg/mL y 0,0011 nmol glucosa/min, respectivamente.By applying different conservation techniques on yellow pitaya fruit (Acanthocereus pitajaya it has been found that excessive softening of the peel contributes to the deterioration of the fruit. Due to that both cellulase and xylanase have been related to the softening of the fruit's peel; this work was based on the search of the best conditions not only for the extraction, but also for the activity measurement of both cellulase and xylanase. The best extraction system for both enzymes was 20 mM buffer phosphate, 0.5 M NaCl, pH 7.0. For the cellulase activity measurement it was necessary to incubate during 60 min at 37 ºC, with a volume of raw enzymatic extract of 30 µL, using buffer acetate 100 mM at pH 5,0; the values of apparent K M and V MÁX were 0.279 mg/mL and 0.00014 nmol glucose/min, respectively. To

  15. 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

  16. Formation of cellulase and protein in the growth of Chaetomium cellulolyticum on cellulose-containing substrates

    Energy Technology Data Exchange (ETDEWEB)

    Faehnrich, P.; Irrgang, K.

    1982-01-01

    C. cellulolyticum was grown on glucose and different types of cellulose. The organism studied was mutant 7S/7, derived from C. cellulolyticum ATCC 32319. Cellulase formation on glucose was very slight, whereas it was much greater on cellulose, indicating that most of the activity is induced. Both cellulose and protein production were greater on newsprint than on Avicel, and greater on alkali-treated than on untreated newsprint. Crystaline structure in cellulose inhibited fermentation.

  17. 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.

    EDTA H 2 O 2 system i.e. fenton reaction [25]. The reaction was performed in phosphate buffer (20 mM, pH 7.4) containing 2– deoxyribose (2.8 mM), FeCl 3 (100 µM), H 2 O 2 (1 mM), EDTA (100 µM) and various concentrations (50–200 µg/ml) of sample... absorbance) × 100] 7 2.4.7. Ferric reducing/antioxidant power (FRAP) assay The FRAP assay was performed for the samples and reference compound of various concentrations (50 – 200 µg/m) based on Benzie and Strain [26]. The FRAP reagent (2.5 ml of 20...

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

    Directory of Open Access Journals (Sweden)

    E. P. Kopylov

    2009-11-01

    Full Text Available 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 glutamine synthetase activity, chlorophylls content in leaves and plants’ productivity.

  19. Production of xylanase and CMCase on solid state fermentation in different residues by Thermoascus aurantiacus miehe Produção de xilanase e CMCase por fermentação em estado sólido em diferentes resíduos pelo fungo termofílico Thermoascus aurantiacus miehe

    Directory of Open Access Journals (Sweden)

    Roberto da Silva

    2005-09-01

    Full Text Available The use of waste as raw material is important for government economy and natural balance. The purpose of this work was to study the production of CMCase and xylanase by a Brazilian strain of Thermoascus aurantiacus in solid state fermentation (SSF using different agricultural residues (wheat bran, sugarcane bagasse, orange bagasse, corncob, green grass, dried grass, sawdust and corn straw as substrates without enrichment of the medium and characterize the crude enzymes.The study of the extracellular cellulolytic and hemicellulolytic enzymes showed that T. arantiacus is more xylanolytic than cellulolytic. The highest levels of enzymes were produced in corncob, grasses and corn straw. All the enzymes were stable at room temperature by 24 h over a broad pH range (3.0-9.0 and also were stable at 60ºC for 1 h. The optimum pH and temperature for xylanase and CMCase were 5.0-5.5 and 5.0 and 75ºC, respectively. The microorganism grew quickly in stationary, simple and low cost medium. The secreted extracellular enzymes presented properties that match with those frequently required in industrial environment.O emprego de residuos como matéria prima é importante como estrategia governamental e para o balanço ambiental. O propósito deste trabalho foi estudar a produção de CMCase e xilanase de uma linhagem de Thermoascus aurantiacus isolado de solo brasileiro em fermentação em estado sólido (SSF usando diferentes resíduos agrícolas (farelo de trigo, bagaço de cana, bagaço de laranja, sabugo de milho, grama verde, grama seca, serragem de eucalipto e palha de milho como substratos sem enriquecimento dos meios e caracterizar as enzimas. O estudo das enzimas hemiceluloliticas extracelulares mostrou que o fungo T. arantiacus é mais xilanolítico do que celulolítico. Ele produziu maiores níveis das enzimas em meios contendo sabugo de milho, grama e palha de milho. Todas as enzimas foram estáveis por 24 h à temperatura ambiente numa ampla faixa

  20. 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.

  1. Resistance determinant erm(X) is borne by transposon Tn5432 in Bifidobacterium thermophilum and Bifidobacterium animals subsp. lactis

    NARCIS (Netherlands)

    Hoek, van A.H.A.M.; Mayrhofer, S.; Domig, K.J.; Aarts, H.J.M.

    2008-01-01

    The erm(X) gene from erythromycin- and clindamycin-resistant Bifidobacterium strains was characterised by polymerase chain reaction and sequence analysis, including flanking regions. Results suggest that the resistance determinant was part of transposon Tn5432 that has been described in several oppo

  2. Prediction of Optimum pH of G/11 Xylanases and the Relationship between the Location of Amino Acid and Optimum pH Value%G/11木聚糖酶最适 pH 值的预测及其与氨基酸位置的关系

    Institute of Scientific and Technical Information of China (English)

    林源清; 张光亚

    2014-01-01

    把木聚糖酶全序列均分为 N 端,中间端(I 端)及 C 端3个部分,并分别以全序列及分段氨基酸的组成作为模型输入值.通过主成分分析(PCA)方法探讨全序列及分段氨基酸组成和最适 pH 值的相关性,运用均匀设计法分别优化支持向量机和 BP 神经网络运行参数.研究结果表明:支持向量机获得的预测模型优于神经网络,其中 RBF 支持向量机是最佳的模型.主成分分析结果显示:I 端主成分跟最适 pH 值相关性最高;相关系数 R 绝对值为0.68,得到的结果与支持向量机结果一致.%We divided the xylanase sequences into three equally segments named N-terminus,I-terminus and C-terminus. And then,we calculated the amino acid compositions of the whole sequences and the segmented sequences,respectively. The amino acid compositions were used as the input values of these models.The principal component analysis (PCA) method was utilized to analyze the relationship between the amino acid composition and the optimum pH.The uniform de-sign was used to optimize the running parameters of support vector machines (SVM)and neural network (BPNN),re-spectively.Our results showed the predicted model obtained by SVM was better than that of BPNN,and the SVM model based on RBF kernel was best.The results of PCA showed the correlation between principle component and optimum pH was best in the I-terminus with the R=-0.68,which coincided with the result of the SVM.

  3. Structure of the Tuberous Sclerosis Complex 2 (TSC2) N Terminus Provides Insight into Complex Assembly and Tuberous Sclerosis Pathogenesis.

    Science.gov (United States)

    Zech, Reinhard; Kiontke, Stephan; Mueller, Uwe; Oeckinghaus, Andrea; Kümmel, Daniel

    2016-09-16

    Tuberous sclerosis complex (TSC) is caused by mutations in the TSC1 and TSC2 tumor suppressor genes. The gene products hamartin and tuberin form the TSC complex that acts as GTPase-activating protein for Rheb and negatively regulates the mammalian target of rapamycin complex 1 (mTORC1). Tuberin contains a RapGAP homology domain responsible for inactivation of Rheb, but functions of other protein domains remain elusive. Here we show that the TSC2 N terminus interacts with the TSC1 C terminus to mediate complex formation. The structure of the TSC2 N-terminal domain from Chaetomium thermophilum and a homology model of the human tuberin N terminus are presented. We characterize the molecular requirements for TSC1-TSC2 interactions and analyze pathological point mutations in tuberin. Many mutations are structural and produce improperly folded protein, explaining their effect in pathology, but we identify one point mutant that abrogates complex formation without affecting protein structure. We provide the first structural information on TSC2/tuberin with novel insight into the molecular function.

  4. 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.

  5. 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.

  6. Identification problems with sterile fungi, illustrated by a keratitis due to a non-sporulating Chaetomium-like species

    NARCIS (Netherlands)

    Vinod Mootha, V.; Shahinpoor, P.; Sutton, D.A.; Xin, L.; Najafzadeh, M.J.; de Hoog, G.S.

    2012-01-01

    A 39-year-old farm worker was injured in her right eye by a piece of wire, which resulted in a corneal ulcer unresponsive to antibiotic treatment. The clinical appearance was that of a corneal infiltrate with feathery borders resembling fungal keratitis. Corneal scrapings were collected and the pati

  7. Production of Agaricus bisporus on substrates pre-colonized by Scytalidium thermophilum and supplemented at casing with protein-rich supplements.

    Science.gov (United States)

    Coello-Castillo, M M; Sánchez, J E; Royse, D J

    2009-10-01

    The objective of this study was to evaluate performance of Agaricus bisporus (Ab) on substrates pre-colonized by Scytalidiumthermophilum (St), a thermophilic fungus known to enhance yields of Ab and increase selectivity of the substrate. The radial extension rate (RER) of the mycelium of three strains of St and their influence on the growth of a brown strain of Ab were evaluated. We also determined the time required for colonization of pangola grass by St in a compost pile and the influence of three protein-rich supplements on yield of Ab on pangola grass (Digitaria decumbens) colonized by St. RER of St ranged from 10.1mm/d on grass to 18.9 mm/d on potato dextrose yeast extract agar, with significant differences among substrates and among strains. Ab grew faster on substrate colonized for 1, 2, or 3 days by St (RER of 3.31, 3.29, 3.23 mm/d, respectively) compared to non-colonized substrate (1.85 mm/d). Ab was cultivated on substrate samples selected daily from the St-inoculated pile, with biological efficiencies (BE) ranging from 4% (day 0) to 73.9% (day 2). Protein-rich supplements (soybean, black beans and cowpeas) added at casing significantly stimulated mushroom yield on St-colonized substrate compared to the non-supplemented control. BE varied from 26.1% on substrate non-supplemented to 73.1% on compost supplemented with ground soybean. There were no significant differences in mushroom yield observed among supplements evaluated.

  8. 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.

  9. LC-MS based analysis of secondary metabolites from Chaetomium and Stachybotrys growth in indoor environments

    DEFF Research Database (Denmark)

    Dosen, Ina

    spores and mycelium parts containing secondary metabolites and bioactive compounds are released from the building material. Not only is the variety of different fungal species present in the indoor environment large, the number of bioactive compounds they produce is also broad. This PhD study focused...... in wet, fungi contaminated indoor environment. Otherwise healthy people may also experience negative health effects, such as skin rashes, headaches, dizziness and chronic fatigue. During their growth on building materials, indoor fungi produce and release many different kinds of components....... These components are suspected of causing adverse health effects, however that causality has yet to be documented. Fungi produce biomass in the form of mycelium and spores both of which contain an array of secondary metabolites and bioactive compounds. When fungal biomass dries up, whole viable spores, fragmented...

  10. Rapid detection and identification of Stachybotrys and Chaetomium species using tissue PCR analysis

    DEFF Research Database (Denmark)

    Lewinska, Anna Malgorzata; Peuhkuri, Ruut Hannele; Rode, Carsten

    2016-01-01

    Indoor fungi are a worldwide problem causing negative health effects for infected building's occupants and even deterioration of building structures. Different fungal species affect buildings and their inhabitants differently. Therefore, rapid and accurate identification of fungi to the species l...

  11. Biocidal synthetic coatings based on high-molecular metaloorganic compounds. [Aspergillus flavus; Aspergillus niger; Aspergillus terreus; Penicillium funiculosum; penicillium cyclopium; Penicillium chrysogenum; Paecilomyces varioti; Chaetomium globosum; trichoderma viride

    Energy Technology Data Exchange (ETDEWEB)

    Mishchenko, V.F.; Zubov, V.A.; Eremenko, Yu.G.

    Long-term stays of man and animals in closed life-support systems lead to contamination of the space-craft by various microorganisms. Organotin compounds are considered promising biocidal agents for paints and varnishes with a broad spectrum of action against a variety of microorganisms. Several organotin polymers of the acrylate type were prepared and were found to be effective fungicides.

  12. 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

    The mature barley grain contains a complement of enzymes that are synthesized during seed development for degradation of seed storage reserves during germination. These enzyme activities (first wave enzymes) are considered important for maximizing nutrient digestibility in food and feed. Several...... of these enzyme activities between barley cultivars, as well as the distribution and composition of the residing commensal fungal community and the grain surface associated proteins. To obtain new insight into the interplay between barley grains and the colonizing fungi, we set out to study the activities...

  13. Evidence that pentosans and xylanase affect the re-agglomeration of the gluten network

    NARCIS (Netherlands)

    Wang, M.; Vliet, T. van; Hamer, R.J.

    2004-01-01

    In the gluten-starch separation process gluten is formed first as a result of breakdown of the gliadin-glutelin structures during mixing followed by their re-agglomeration. To date the effect of pentosans and enzymes have not been studied separately. A simple modification of TNO Glutomatic system en

  14. Assessment of xylanase activity in dry storage as a potential method of reducing feedstock cost.

    Science.gov (United States)

    Smith, William A; Thompson, David N; Thompson, Vicki S; Radtke, Corey W; Carter, Brady

    2009-05-01

    Enzymatic preprocessing of lignocellulosic biomass in dry storage systems has the potential to improve feedstock characteristics and lower ethanol production costs. To assess the potential for endoxylanase activity at low water contents, endoxylanase activity was tested using a refined wheat arabinoxylan substrate and three commercial endoxylanases over the water activity range 0.21-1.0, corresponding to water contents of 5% to >60% (dry basis). Homogeneously mixed dry samples were prepared at a fixed enzyme to substrate ratio and incubated in chambers at a variety of fixed water activities. Replicates were sacrificed periodically, and endoxylanase activity was quantified as an increase in reducing sugar relative to desiccant-stored controls. Endoxylanase activity was observed at water activities over 0.91 in all enzyme preparations in less than 4 days and at a water activity of 0.59 in less than 1 week in two preparations. Endoxylanase activity after storage was confirmed for selected desiccant-stored controls by incubation at 100% relative humidity. Water content to water activity relationships were determined for three lignocellulosic substrates, and results indicate that two endoxylanase preparations retained limited activity as low as 7% to 13% water content (dry basis), which is well within the range of water contents representative of dry biomass storage. Future work will examine the effects of endoxylanase activity toward substrates such as corn stover, wheat straw, and switchgrass in low water content environments.

  15. 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...

  16. Dicty_cDB: Contig-U04414-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ... 44 8.6 1 ( DV547508 ) rbcmb0_000208 Chaetomium cupreum mycelium cDNA li... 44 8.6 1 ( DV547506 ) rbcmb0_...000206 Chaetomium cupreum mycelium cDNA li... 44 8.6 1 ( DV547503 ) rbcmb0_000203 Chaetomium cupreum mycel...ium cDNA li... 44 8.6 1 ( DV547502 ) rbcmb0_000202 Chaetomium cupreum mycelium cDNA... li... 44 8.6 1 ( DV547441 ) rbcmb0_000101 Chaetomium cupreum mycelium cDNA li... 44 8.6 1 ( DB915235 ) Idio

  17. 产耐高温木聚糖酶菌株的筛选及其产酶条件优化%Screening of thermostable xylanase producing strains and its condition optimization for xylanase production

    Institute of Scientific and Technical Information of China (English)

    孙明哲; 郑宏臣; 孙君社; 裴海生; 刘逸寒; 张璟; 韩杨; 路福平

    2013-01-01

    从牛粪堆肥中分离、筛选到1株产耐高温木聚糖酶的细菌NF1,经形态特征和16S rDNA序列分析,鉴定菌株NF1为类芽孢杆菌,命名为类芽孢杆菌(Paenibacillus sp.) NF1.采用响应面法对该菌种产木聚糖酶的发酵条件进行优化.首先利用Plackett Burman试验设计筛选出影响产酶量的3个主要因素,即牛肉膏浓度、NaCl浓度和初始pH值.在此基础上使用Design-Expert软件进行Box-Behnken试验设计,通过响应面分析得出菌株NF1发酵产酶的最佳条件为玉米芯20g/L、牛肉膏28.2g/L、K2HPO45g/L、MgSO4 0.5g/L、NaC1 7.1 g/L、初始pH值为6.9;装液量40mL、转速160r/min、接种量2%、温度28℃.经过验证试验,优化后粗酶液酶活达到160.6IU/mL,与响应面预测结果一致,较优化前木聚糖酶酶活提高了51.4倍.

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

    DEFF Research Database (Denmark)

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

    2010-01-01

    . Extensive analysis of the cell walls showed a 10%-15% increase in arabinose to xylose ratio, a 50% increase in the proportion of water-extractable arabinoxylan, and a shift in the MW of the water-extractable arabinoxylan from being mainly larger than 85 kD to being between 2 and 85 kD. Ferulic acid esterase......-expressing grains were also shrivelled, and the seed weight was decreased by 20%-50%. No ferulic acid esterase activity could be detected in wild-type grains whereas ferulic acid esterase activity was detected in transgenic lines. The grain cell walls had 15%-40% increase in water-unextractable arabinoxylan...

  19. Thermal stability of xylanases produced by Aspergillus awamori Estabilidade térmica de xilanases produzidas por Aspergillus awamori

    Directory of Open Access Journals (Sweden)

    Judith Liliana Solórzano Lemos

    2000-09-01

    Full Text Available The effect of temperature on the activity and stability of endoxylanase and beta-xylosidase from Aspergillus awamori was investigated. The growth of A. awamori in milled sugar cane bagasse produced predominantly extracellular endoxylanase (30 U/ml and lower amounts of beta-xylosidase (1.3 U/ml. Grown in sugar cane bagasse as the principal carbon source, the microorganism produced a quite stable beta-xylosidase in a temperature range of 35-55°C, but it exhibited a lower thermostable endoxylanase. The thermostability of endoxylanase was enhanced through addition of polyhydric alcohols, mainly 2 M xylitol and sorbitol solutions. Particular stability upon storage (100% was found for endoxylanase at -4°C for 165 days. Yet for beta-xylosidase, an activity decrease of approximately 20% was observed during the first 15 days of storage, maintaining roughly 75% of initial activity until the end of the experiment.O presente trabalho trata do estudo do efeito da temperatura na atividade e estabilidade de endo-xilanase e beta-xilosidase produzidas, extracelularmente, por Aspergillus awamori. O cultivo deste microrganismo, em bagaço de cana finamente dividido, produziu predominantemente endo-xilanases (30 U/ml e menores atividades de beta-xilosidase (1,3 U/ml; esta última exibiu considerável estabilidade em faixa de temperatura variando de 35 a 55°C, por outro lado verificou-se uma menor termo estabilidade para a endoenzima. A estabilidade térmica de endo-xilanase foi aumentada consideravelmente através da adição de polióis, principalmente xilitol e sorbitol em concentração de 2,0 M. No que concerne à estocagem a baixa temperatura (-4°C, observou-se uma estabilidade particular na atividade endo-xilanásica (100%, durante 165 dias, porém, um decréscimo de aproximadamente 20% na atividade beta-xilosidásica foi verificado após os primeiros 15 dias de armazenamento nas mesmas condições, mantendo-se aproximadamente em 75% da atividade inicial no mesmo período de tempo.

  20. Studies on the Xylanase Producer Aspergillus niger%黑曲霉产木聚糖酶的研究

    Institute of Scientific and Technical Information of China (English)

    张年凤; 赵允麟

    2003-01-01

    筛选了1株高产木聚糖酶的黑曲霉(Aspergillus niger)An-238菌株,研究了其在固态培养基中的产酶条件.该菌株发酵曲中除含有木聚糖酶5 117 U/g(干曲)外,还有纤维素酶425 U/g(干曲),果胶酶1 236 U/g(干曲),蛋白酶22 531 U/g(干曲).

  1. 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.

  2. Dicty_cDB: Contig-U04139-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available N PROGRESS *** f... 38 8.8 2 ( DV547337 ) rbcma0_005360 Chaetomium cupreum mycelium cDNA li... 34 9.1 2 ( DV...547434 ) rbcma0_005590 Chaetomium cupreum mycelium cDNA li... 34 9.1 2 ( BI067048 ) pgf1n.pk010.h1 normalize

  3. 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.

  4. 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

    heat-treatment and soaking time (P≤0.001). This was mainly due to a smaller proportion of non-degraded InsP6-P at 24 h in HW compared with NHW (0.13 vs. 0.47) (P≤0.001) possibly caused by structural changes imposed by the heat-treatment. In SBM, RSC, SBM/NHW or RSC/NHW, the InsP6 degradation...... in a negligible accumulation of InsP5-InsP2. Soaking of plant feedstuffs prior to feeding of pigs may improve P digestibility compared with dry feeding, but studies with pigs are required to clarify this....

  5. 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

    with CAX, EX, or EXP—in a double 4 × 4 Latin square design. The experimental period lasted 7 d; adaptation lasted 4 d, and the ileal digesta were collected for 8 h on d 5 and 7, when spot samples of feces were also collected. Digesta samples were analyzed for NDC, total and soluble nonstarch...

  6. 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

    in the broiler caeca were Clostridium coccoides-Eubacterium rectale followed by Bacteroides sp., Lactobacillus sp./Enterococcus sp., Bifidobacterium sp. and Enterobacteriaceae. For both cereal types, the enzyme supplementation significantly decreased the relative amount of Enterobacteriaceae. 4. The T...

  7. 木聚糖酶的分类及其在面包烘焙中的应用%Xylanase Families and its Application in Baking Industry

    Institute of Scientific and Technical Information of China (English)

    聂文秀

    2013-01-01

    木聚糖酶(E.C.3.2.1.8)是一类重要的降解线性β-1,4-木聚糖生成木糖、木二糖或低木聚糖等的木糖苷水解酶,并因其巨大的工业潜在价值而得到越来越多研究者的关注.本文详尽地对目前被发现的木聚糖酶进行家族分类研究,阐述各家族成员水解特点,并综述了木聚糖酶在面包烘焙中的应用进展.

  8. 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.

  9. 黑曲霉产木聚糖酶发酵条件的研究%The Study on Xylanase fermentation by Aspergillus niger sp.

    Institute of Scientific and Technical Information of China (English)

    毕瑞明; 孙迅; 任少亭

    2000-01-01

    正交设计试验结果表明,黑曲霉(Aspergillus niger m12)产木聚糖酶活力达76.60 u/ml,合适的产酶发酵条件如下,培养基(g/L):麸皮40,尿素6.67,KH2PO41.0,MgSO4·7H2O 0.5,NaCl 0.3,Tween-80 3.0,CaCO3 2.0,28℃,120 r/min水浴振荡培养5.5 d.

  10. 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...

  11. Fungal bis-Naphthopyrones as Inhibitors of Botulinum Neurotoxin Serotype A

    Science.gov (United States)

    2012-04-02

    Chaetochromin A (1) was isolated from solid-substrate fermentation cultures of Chaetomium arcuatum (syn. Chaetomium virescens) (NRRL 25243 = IMI...86456) isolated from a soil sample collected in Lucknow, India.21 Talaroderxines A and B (2 and 3) were obtained from liquid cultures of a coprophilous... feeding chaetochromin-containing diet. Proc. Jpn. Assoc. Mycotoxicol. 1982, 22−23. (35) Ito, Y.; Ohtsubo, K. Teratogenicity of oral chaetochromin, a

  12. 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; Triticum Aestivum Molecule: Xylanase Inhibitor Protein I; Chain: A; Synonym: Xip-1; Molecule: Endo-1,4-Xylanase; Chain...: B; Synonym: Gh11; Engineered: Yes Hydrolase Inhibitor/Hydrolase 3.2.1.8 (Endo-1,4-Xylanase...rand, P.Manzanares, H.J.Gilbert, N.Juge, A.Roussel The Dual Nature Of The Wheat Xylanase Protein Inhibitor X

  13. Development of a Genome-Proxy Microarray for Profiling Marine Microbial Communities and its Application to a Time Series in Monterey Bay, California

    Science.gov (United States)

    2008-09-01

    Costas, A.M., Maresca, J.A., Chew, A.G., Klatt, C.G., Bateson , MM. et al. (2007) Candidatus Chloracidobacterium thermophilum: an aerobic...Microbiology 70: 4103-4110. Thyssen, M., Tarran, G.A., Zubkov, M.V., Holland, R.J., Gregori , G., Burkill, P.H., and Denis, M. (2008) The emergence of

  14. Solid-state fermentation conditions of xylanase by Thermophilic sporotrichum%嗜热侧孢霉固体发酵产木聚糖酶条件研究

    Institute of Scientific and Technical Information of China (English)

    张辉; 孙占斌; 桑青

    2010-01-01

    对嗜热侧孢霉(Thermophilic sporotrichum)木聚糖酶条件进行了研究.经正交试验得m,最佳产酶条件为:玉米芯+麸皮(1:2)15.0g,(NH4)2SO4+蛋白胨(2:1)2%,KH2PO40.3%,MgSO4·7H2O0.05%,pH6.0,含水量60%,培养温度和时间分别为40℃和168h.在此培养条件下木聚糖酶活力为5633.2 U.

  15. 添加绿汁发酵液和木聚糖酶对稻草青贮品质的影响%Effects of Fermented Green Juice and Xylanase on Quality of Rice Straw Silage

    Institute of Scientific and Technical Information of China (English)

    朱小清; 邹长连; 陈国富; 叶杭; 张文昌; 庄益芬

    2014-01-01

    本试验旨在研究绿汁发酵液和木聚糖酶对稻草青贮品质的影响.在常规水分(MC1)稻草青贮和低水分(MC2)稻草青贮中均设对照(CON)组及添加2 mL/kg绿汁发酵液(FGJ)组、20 mg/kg木聚糖酶(XYL)组、2 mL/kg绿汁发酵液与20 mg/kg木聚糖酶的复合(MIX)组,每个处理组重复3次.常温下贮存60 d开封,测定青贮的发酵品质和化学成分.结果表明,MC1青贮中,与CON组相比,MIX组水分和酸性洗涤纤维(ADF)含量显著降低(P<0.05),FGJ、XYL、MIX组氨态氮(AN)含量极显著降低(P<0.01)、水溶性碳水化合物(WSC)含量显著或极显著增加(P<0.05;P<0.01),XYL、MIX组中性洗涤纤维(NDF)含量显著降低(P<0.05),FGJ组pH极显著降低(P<0.01);MC2青贮中,与CON组相比,FGJ组水分含量显著降低、水溶性碳水化合物含量及干物质回收率(DMR)显著增加(P<0.05),FGJ、MIX组氨态氮和中性洗涤纤维含量显著或极显著降低(P<0.05;P<0.01)、pH极显著降低(P<0.01),MIX组酸性洗涤纤维含量显著降低(P<0.05).较MC1青贮,MC2青贮的pH和干物质回收率极显著增加(P<0.01),水溶性碳水化合物含量显著增加(P<0.05),氨态氮含量极显著降低(P<0.01).综上所述,在常规水分和低水分稻草青贮中,3种添加剂均有显著的添加效果,以绿汁发酵液与木聚糖酶复合添加的效果最佳,且低水分稻草青贮的效果优于常规水分稻草青贮.

  16. 小麦型日粮中添加木聚糖酶对肉鸭生产性能的影响%Effect of xylanase supplementation in wheat based diet on the growth performance of meat ducks

    Institute of Scientific and Technical Information of China (English)

    杨加豹; 张滴; 魏敏

    2010-01-01

    本试验在小麦型基础日粮中分别添加木聚糖酶0、50、100、150、200g/t饲料(5000U/g酶)饲喂12日龄樱桃谷肉鸭,观察木聚糖酶对肉鸭生产性能的影响.结果表明:添加木聚糖酶对肉鸭生产性能影响不显著(P>0.05),单位增重饲料成本以添加量50 g/t(即每1 kg饲料含250U木聚糖酶)的有效含量为佳.

  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

    24 h at pH 5, 50 degrees C. A 10%:40%:50% mixture of Abf II, Abf III, and beta-xyl released 56 mg of arabinose and 91 mg of xylose per gram of vinasse dry matter after 24 h at pH 5, 50 degrees C. The optimal dosages of the "minimal" enzyme cocktails were determined to be 0.4, 0.3, and 0.2 g enzyme......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...

  18. 米曲霉RIBl28耐酸性木聚糖酶的研究%Study on An Acid-Stable Xylanase from Aspergillus oryzae RIB128

    Institute of Scientific and Technical Information of China (English)

    陆健; 曹钰; 陈坚; 若林三郎

    2001-01-01

    通过RBB-Xylan筛选平板和固体培养方法获得了产木聚糖酶菌株米曲霉(Aspergillus oryzae) RIBl28.在液体发酵时木聚糖是有效的诱导底物.通过离子交换和凝胶过滤色谱纯化得到了一个耐酸性木聚糖酶(木聚糖酶B).它的相对分子质量为65 000,最适作用温度和pH分别为55 ℃(pH 6.5),它在50 ℃时很稳定,并且在pH 2.0时仍保留相当高的活性.木聚糖酶B含有较多的天冬氨酸、丝氨酸、苏氨酸和丙氨酸.

  19. Combinative Degradation of Xylan with Acetyl Xylan Esterase and Xylanase%乙酰木聚糖酯酶协同木聚糖酶降解木聚糖的研究

    Institute of Scientific and Technical Information of China (English)

    2016-01-01

    探讨了宇佐美曲霉(A spergillus usamii)乙酰木聚糖酯酶和第10、11家族木聚糖酶之间的协同作用.利用作者所在实验室构建保藏的3株工程酵母Pichia pastoris GS115/A uaxe、GS115/Auxyn11A和GS115/A uxyn10A进行甲醇诱导发酵,分别获得重组乙酰木聚糖酯酶和木聚糖酶.在木聚糖酶最适pH、40℃、料液质量体积比1 g:60 mL、水解2h的条件下分别研究了不同加酶量作用于小麦麸皮时生成的还原糖量.结果表明,乙酰木聚糖酯酶与第11家族木聚糖酶有更好的协同作用,并在添加量比(酶活力比)为5:1时所测协同效果最好,还原糖生成量较木聚糖酶单独作用增加了46%.因此,在乙酰木聚糖酯酶的作用下,木聚糖上乙酰基的去除,对提高木聚糖酶对木聚糖的水解效率具有重要作用.

  20. 木聚糖酶产生菌的分离筛选及酶学性质%Separation and Selection to Bacteria Strains Producting Xylanase and Their Enzymatic Properties

    Institute of Scientific and Technical Information of China (English)

    葛晓萍; 郭清吉; 石琰璟; 冯灵刚

    2007-01-01

    利用透明圈法从云南腾冲温泉出水口土样中粗筛出12株木聚糖酶产生茵,其中Xylan-12菌活力较高,对其进行了16S rDNA部分序列测定,经BLAST同源性分析确定其为Bacillus flavothermus.对Xylan-12产生的木聚糖酶进行了酶学性质的初步研究,结果表明,该酶的最适PH为6,最适温度为65℃.

  1. Cloning of Acidic Xylanase Gene and Its Secretion Expression in Pichia pastoris%酸性木聚糖酶基因的克隆及其在毕赤酵母中的分泌表达

    Institute of Scientific and Technical Information of China (English)

    李春华; 李翔; 马立新

    2005-01-01

    运用"鸟枪法"克隆构建了环境微生物的基因组文库,并从中筛选得到一个酸性木聚糖酶基因,命名为xyl3,其在GenBank中的登录号为gb:AY300805.BLAST分析表明,该基因的序列同源性很低,其中仅存在很短的木聚糖酶基因的同源片段,其编码的木聚糖酶属于Glycosyl hydrolases family 10,与来源于Geobacillus stearothermophilus的intra-cellularxylanase在氨基酸水平具77%同源性.该基因经T4 DNA polymerase处理后,克隆至经限制性内切酶Cpo Ⅰ和Not Ⅰ双酶切后的毕赤酵母表达载体pHBM905,获得重组质粒pHBM706.此重组质粒转化毕赤酵母GS115,经含有交联木聚糖的选择性培养平板和PCR扩增鉴定筛选得到重组毕赤酵母GS115(pHBM706).以0.5%甲醇于28℃诱导产酶,测得重组毕赤酵母GS115(pHBM706)在诱导的第36h产酶达最高值,所产粗酶液酶活为0.177 IU/mL.该酶的最适反应pH为5.5,最适反应温度为50℃.

  2. Studies on the influencing factors for xylanase production by Pleurotus ostreatus SYJ-012%食用菌Pleurotus ostreatus SYJ042产木聚糖酶的影响因素研究

    Institute of Scientific and Technical Information of China (English)

    岳晓禹; 蔡青和; 牛天贵; 惠明; 隋继学; 张雪

    2006-01-01

    实验研究了食用菌SYJ042在液体培养条件下,不同的培养条件对该菌产木聚糖酶的影响.研究表明,培养条件(碳源、氮源、通气量、初始pH和接种量)对其产木聚糖酶有显著的影响.最佳碳源为2.5%玉米芯+2.5%麸皮,氮源为0.2%的蛋白胨,接种量每50mL发酵液加4块菌丝块,通气量为发酵液占容积的1/3,初始pH6.0.

  3. 平菇产木聚糖酶固态发酵条件优化和酶学性质研究%Study on enzymatic characteristics and solid-phase fermentation condition of xylanase from Pleurotus ostreatus

    Institute of Scientific and Technical Information of China (English)

    吴萍; 周鸣鸣; 盛伟

    2010-01-01

    利用平菇(Pleurotus ostreatus)降解甘蔗渣固态发酵生产木聚糖酶,通过单因素和正交实验确定最佳培养基组分及其配比,并时其粗酶的酶学性质进行了研究.结果表明:碳源为甘蔗渣8g:玉米芯2g,氮源为2.0%酵母膏,pH为4,料水比为1:3.2,装瓶量为5g/125mL三角瓶,培养10d时,得到的酶活力最高,产酶量可达2918.95 U/g.粗酶液的最适反应pH为5,最适反应温度为40℃,在pH4~6的范围内酶活性较稳定.温度的适应性较宽,在30~70℃的范围里,相对酶活仍保持在65%以上.

  4. 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.

  5. 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.

  6. Studies on the Production of Xylanase for Submerged Culture by Aspergillus Clavatus GM-69%棒曲霉GM-69深层发酵生产木聚糖酶的研究

    Institute of Scientific and Technical Information of China (English)

    彭中健; 翁照南; 梁淑娃; 谭颖嫦; 蓝碧锋; 杜少平; 陈郧东

    2004-01-01

    报道了发酵条件对棒曲霉GM-69产木聚糖酶的影响,并用50L发酵罐进行了深层发酵试验.确定产酶摇瓶培养的最佳条件:培养时间为72h,温度为28℃,培养基起始pH为3.8,接种量为5%(V/V),转速为250r/min,装液量为90mL/500mL三角瓶.其酶活力最高362IU/mL,平均为343 IU/mL,比培养条件优化前增加了55.6%.50L发酵罐发酵最佳条件:转速为400r/min,通风量为1:0.6~1,罐压为0.08Mpa,装量为70%,发酵周期为72h,温度28℃,培养基起始pH3.8,接种量5%(V/V),酶活力平均为351IU/mL.

  7. 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.

  8. Five new records of thermotolerant fungi from China%耐热真菌五个中国新记录种

    Institute of Scientific and Technical Information of China (English)

    张勇; 李多川

    2013-01-01

    Based on morphological characteristics and the sequence of ribosomal DNA-ITS, five new records of thermotolerant fungi are reported, they are: Trichoderma ghanense, Neosartorya fennelliae, Hypoxylon fragiforme, Chaetomium jodhpurense, Chaetomium luteum. Illustrated descriptions of the species were given according to the Chinese strains, and the taxonomy of these species was discussed. Specimens and living cultures examined are deposited in the Herbarium of Shandong Agricultural University, Plant Pathology (HSAUP).%通过形态特征观察、rDNA-ITS序列测定及分析,报道5个耐热真菌中国新记录种,即加纳木霉Trichoderma ghanense,芬尼新萨托菌Neosartorya fennelliae,草莓状炭团菌Hypoxylon fragiforme,焦特普尔毛壳Chaetomium jodhpurense,黄色毛壳Chaetomium luteum.根据所采集的标本和菌种对这些种进行了描述和讨论.研究标本保存在山东农业大学植物病理学标本室(HSAUP).

  9. Farrowia, a new genus in the Chaetomiaceae

    NARCIS (Netherlands)

    Hawksworth, D.L.

    1975-01-01

    The new genus Farrowia D. Hawksw. is described to accommodate Chaetomium longicolleum Krzem. & Badura and C. longirostre (Farrow) L. Ames, species formerly incorrectly referred to Chaetoceratostoma Turc. & Maffei. These two species are united under the name F. longicollea (Krzem. & Badura) D. Hawksw

  10. Associations between Fungal Species and Water-Damaged Building Materials

    DEFF Research Database (Denmark)

    Andersen, Birgitte; Frisvad, Jens Christian; Søndergaard, Ib;

    2011-01-01

    melleus, Aspergillus niger, Aspergillus ochraceus, Chaetomium spp., Mucor racemosus, Mucor spinosus, and concrete and other floor-related materials. These results can be used to develop new and resistant building materials and relevant allergen extracts and to help focus research on relevant mycotoxins...

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

    NARCIS (Netherlands)

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

    2007-01-01

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

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

    NARCIS (Netherlands)

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

    2001-01-01

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

  13. 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.

  14. The effect of the introduction of exogenous strain Acidithiobacillus thiooxidans A01 on functional gene expression, structure and function of indigenous consortium during pyrite bioleaching.

    Science.gov (United States)

    Liu, Yi; Yin, Huaqun; Zeng, Weimin; Liang, Yili; Liu, Yao; Baba, Ngom; Qiu, Guanzhou; Shen, Li; Fu, Xian; Liu, Xueduan

    2011-09-01

    Acidithiobacillus thiooxidans A01 was added to a consortium of bioleaching bacteria including Acidithiobacilluscaldus, Leptospirillumferriphilum, Acidithiobacillus ferrooxidans, Sulfobacillus thermosulfidooxidans, Acidiphilium spp., and Ferroplasma thermophilum cultured in modified 9 K medium containing 0.5% (w/v) pyrite, and 10.7% increase of bioleaching rate was observed. Changes in community structure and gene expression were monitored with real-time PCR and functional gene arrays (FGAs). Real-time PCR showed that addition of At. thiooxidans caused increased numbers of all consortium members except At. caldus, and At. caldus, L. ferriphilum, and F. thermophilum remained dominant in this community. FGAs results showed that after addition of At. thiooxidans, most genes involved in iron, sulfur, carbon, and nitrogen metabolisms, metal resistance, electron transport, and extracellular polymeric substances of L. ferriphilum, F. thermophilum, and Acidiphilium spp., were up-regulated while most of these genes were down-regulated at 70-78 h in At. caldus and up-regulated in At. ferrooxidans, then down-regulated at 82-86 h.

  15. The identification, purification, and characterization of STXF10 expressed in Streptomyces thermonitrificans NTU-88.

    Science.gov (United States)

    Cheng, Hsueh-Ling; Tsai, Chih-Yun; Chen, Hui-Jye; Yang, Shang-Shyng; Chen, Yo-Chia

    2009-03-01

    Multiple xylanolytic enzymes of Streptomyces thermonitrificans NTU-88 were induced by oat-spelt xylan and separated by two-dimensional polyacrylamide and zymogram gels. Nineteen clear spots differed in pI and molecular weight values were found on the zymogram, and only spot one was seen on the corresponding silver-stained gel. These results revealed that multiple xylanases were secreted when S. thermonitrificans NTU-88 was induced and the spot (STXF10), identified as being a glycosyl hydrolase family 10 xylanase, was the predominant one among xylanases. STXF10 showed a tolerance for high temperatures and broad pH ranges and high affinity and hydrolysis efficiency for xylans. Furthermore, it also featured the minor ability to degrade different lignocellulosic substrates. Although S. thermonitrificans NTU-88 possesses multiple xylanases, our results suggest that the major form of xylanase might be selectively and specifically induced depending on the type of substrate to which the microorganism is exposed.

  16. Synergy between cellulases and pectinases in the hydrolysis of hemp.

    Science.gov (United States)

    Zhang, Junhua; Pakarinen, Annukka; Viikari, Liisa

    2013-02-01

    The impact of pectinases in the hydrolysis of fresh, steam-exploded and ensiled hemp was investigated and the synergy between cellulases, pectinases and xylanase in the hydrolysis was evaluated. About half; 59.3% and 46.1% of pectin in the steam-exploded and ensiled hemp, respectively, could be removed by a low dosage of pectinases used. Pectinases were more efficient than xylanase in the hydrolysis of fresh and ensiled hemp whereas xylanase showed higher hydrolytic efficiency than the pectinase preparation used in the hydrolysis of steam-exploded hemp. Clear synergistic action between cellulases and xylanase could be observed in the hydrolysis of steam-exploded hemp. Supplementation of pectinase resulted in clear synergism with cellulases in the hydrolysis of all hemp substrates. Highest hydrolysis yield of steam-exploded hemp was obtained in the hydrolysis with cellulases and xylanase. In the hydrolysis of ensiled hemp, the synergistic action between cellulases and pectinases was more obvious for efficient hydrolysis.

  17. 中国耐热真菌三个新记录种%Three new records of thermotolerant fungi from China

    Institute of Scientific and Technical Information of China (English)

    张勇; 李多川

    2011-01-01

    报道3个耐热真菌中国新记录种,瘤突毛壳Chaetomium strumarium,耐热梭孢壳Thielavia subthermophila,榛色钩囊菌Hamigera avellanea.根据所采集的标本和菌种对这些种进行了描述和讨论.研究标本保存在山东农业大学植物病理学标本室(HSAUP).%Three new Chinese records of thermotolerant fungi are reported,they are Chaetomium strumarium,Thielavia subthermophila and Hamigera avellanea.Illustrated descriptions of the species were given according to our strains,and their taxonomy was discussed.Specimens and living cultures examined arc deposited in the Herbarium of Shandong Agricultural University,Plant Pathology(HSAUP).

  18. 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.

  19. The Effects of Wheat Bran Composition on the Production of Biomass-Hydrolyzing Enzymes by Penicillium decumbens

    Science.gov (United States)

    Sun, Xianyun; Liu, Ziyong; Qu, Yinbo; Li, Xuezhi

    The effects of the starch, protein, and soluble oligosaccharides contents in wheat bran on the extracellular biomass-hydrolyzing enzymes activities released by Penicillium decumbens mycelia grown in batch fermentations have been examined. The results showed increased starch content correlated directly with an increase in released amylase activity but inversely with the levels of secreted cellulase and xylanase. High amounts of protein in wheat bran also reduced the activities of cellulase, xylanase and protease in the culture medium. The effects of the soluble and insoluble components of wheat bran and cello-oligosaccharides supplements on production of extracellular cellulase and xylanase were compared. The soluble cello-oligosaccharides compositions in wheat bran were proved to be one of the most significant factors for cellulase production. According to the results of this research, determining and regulating the composition of wheat bran used as a fermentation supplement may allow for improved induction of cellulase and xylanase production.

  20. An efficient treatment for detoxification process of cassava starch by plant cell wall-degrading enzymes.

    Science.gov (United States)

    Sornyotha, Somphit; Kyu, Khin Lay; Ratanakhanokchai, Khanok

    2010-01-01

    The objective of this work was to remove linamarin in starch from cassava (Manihot esculenta Crantz cv. KU-50) roots, a high-cyanogen variety by using plant cell wall-degrading enzymes, xylanase and cellulase. The combination of xylanase from Bacillus firmus K-1 and xylanase and cellulase from Paenibacillus curdlanolyticus B-6 at the ratio of 1:9 showed the maximum synergism at 1.8 times for hydrolyzing cassava cortex cell walls and releasing linamarase. Combined enzyme treatment enhanced linamarin liberation from the parenchyma by 90%. In addition, when the combined enzymes were applied for detoxification during cassava starch production, a low-cyanide-product was obtained with decreased linamarin concentration (96%) compared to non-enzyme treated tissues. Based on these results, xylanase and cellulase treatment is a good method for low-cyanide-cassava starch production and could be applied for detoxification of cassava products during processing.

  1. 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

    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...

  2. 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

    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...

  3. 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.

  4. Acceleration of Fibrous Biomass Degradation by Bacterial Enzymes

    OpenAIRE

    大宮, 邦雄; 河津, 哲; 孫, 嘉琳; 木村, 哲哉; 苅田, 修一; 粟冠, 和郎; Ohmiya,Kunio; Kawazu, Tetsu; Sun, Jialin; KIMURA, TETSUYA; Karita, Shuichi; Sakka, Kazuo

    1997-01-01

    Since biomass photosynthesized from C02 and H2O is one of the most predominant storage sites of solar energy, its effective utilization will be essential to overcome the shortage of foods and energy in future. Relaxation of biomass tissue is focused to enhance solubilization by expressing fiber‐degrading enzyme genes in plants. A xylanase gene from Clostridium thermocellum was highly expressed in tobacco plant (4%) without apparent defects in the growth. Another xylanase from Clostridium ste...

  5. Thermomyces lanuginosus: properties of strains and their hemicellulases.

    Science.gov (United States)

    Singh, Suren; Madlala, Andreas M; Prior, Bernard A

    2003-04-01

    The non-cellulolytic Thermomyces lanuginosus is a widespread and frequently isolated thermophilic fungus. Several strains of this fungus have been reported to produce high levels of cellulase-free beta-xylanase both in shake-flask and bioreactor cultivations but intraspecies variability in terms of beta-xylanase production is apparent. Furthermore all strains produce low extracellular levels of other hemicellulases involved in hemicellulose hydrolysis. Crude and purified hemicellulases from this fungus are stable at high temperatures in the range of 50-80 degrees C and over a broad pH range (3-12). Various strains are reported to produce a single xylanase with molecular masses varying between 23 and 29 kDa and pI values between 3.7 and 4.1. The gene encoding the T. lanuginosus xylanase has been cloned and sequenced and is shown to be a member of family 11 glycosyl hydrolases. The crystal structure of the xylanase indicates that the enzyme consists of two beta-sheets and one alpha-helix and forms a rigid complex with the three central sugars of xyloheptaose whereas the peripheral sugars might assume different configurations thereby allowing branched xylan chains to be accepted. The presence of an extra disulfide bridge between the beta-strand and the alpha-helix, as well as to an increase in the density of charged residues throughout the xylanase might contribute to the thermostability. The ability of T. lanuginosus to produce high levels of cellulase-free thermostable xylanase has made the fungus an attractive source of thermostable xylanase with potential as a bleach-boosting agent in the pulp and paper industry and as an additive in the baking industry.

  6. 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.

  7. Fungal community dynamics and driving factors during agricultural waste composting.

    Science.gov (United States)

    Yu, Man; Zhang, Jiachao; Xu, Yuxin; Xiao, Hua; An, Wenhao; Xi, Hui; Xue, Zhiyong; Huang, Hongli; Chen, Xiaoyang; Shen, Alin

    2015-12-01

    This study was conducted to identify the driving factors behind fungal community dynamics during agricultural waste composting. Fungal community abundance and structure were determined by quantitative PCR and denaturing gradient gel electrophoresis analysis combined with DNA sequencing. The effects of physico-chemical parameters on fungal community abundance and structure were evaluated by least significant difference tests and redundancy analysis. The results showed that Cladosporium bruhnei, Hanseniaspora uvarum, Scytalidium thermophilum, Tilletiopsis penniseti, and Coprinopsis altramentaria were prominent during the composting process. The greatest variation in the distribution of fungal community structure was statistically explained by pile temperature and total organic carbon (TOC) (P composting.

  8. 中国吐鲁番两株产木聚糖酶的极端耐碱Bacillus halodurans的分类鉴定%Isolation and identification of two xylanase-producing extremely alkali-tolerant strains of Bacillus halodurans from Turpan in China

    Institute of Scientific and Technical Information of China (English)

    易霞; 谢周杰; 邓爱华; 王宁; 艾尔肯·热合曼

    2006-01-01

    通过生理生化实验、16S rDNA 序列分析和同源性杂交,将分离到的XJU-1和XJU-80菌种进行了分类鉴定.XJU-1和XJU-80具有较宽的pH生长范围(分别是pH4.5~12.6和pH3.8~12.6),其G+C mol%含量分别是40.5mol%和42.2mol%.16S rDNA 序列分析和DNA-DNA同源杂交结果表明,XJU-1和XJU-80与Bacillus halodurans C-125和Bacillus halodurans DSM497T具有较高的同源性(99%);两者之间也具有85%的相关性,但其与Bacillus halodurans C-125和Bacillus halodurans DSM497T分别具有81.3%和71.5%的相关性.基于以上结果,将两株分离菌株分类为Bacillus halodurans的两个新品系.%Bacillus halodurans XJU-1 and XJU-80 were characterized in terms of physiological and biochemical characteristics, and 16S rDNA sequence homology and DNA-DNA hybridization analysis. The two isolates can grow in nutrient broth at a broad range of pH values from 4.5 to 12.6 for XJU-1 and from 3.8 to 12.8 for XJU-80, respectively. And the optimum temperature of growth were around 39℃and 42℃, respectively. Phylogenetic analysis of the two strains based on comparison of 16S rRNA sequence revealed that they are closely related to Bacillus halodurans C-125 and DSM497Twith 99% identity. DNA-DNA hybridization showed that the highest levels of DNA-DNA relatedness were found between the two strains (85%) and the B. Halodurans type strains (81.3% and 71.5%), respectively. Moreover, the G+C content of the genomic DNA was 40.5 mol% for XJU-1 and 42.2 mol% for XJU-80.Our results demonstrate that strains XJU-1 and XJU-80 should be classified as two new members of the species B. Halodurans.

  9. 重组枯草芽孢杆菌产极端耐热木聚糖酶条件的优化%Optimization of extreme-thermostable xylanase B production by Bacillus subtilis-xyB in liquid state fermetation

    Institute of Scientific and Technical Information of China (English)

    张伟; 娄恺; 李冠

    2009-01-01

    对基因工程菌Bacillus subtilis-xyB在摇瓶发酵水平产极端耐热木聚糖酶条件进行了优化.结果表明:该基因工程菌发酵产极端耐热木聚糖酶的优化后的培养基配方是:4%麸皮、1%(NH4)2HPO4、0.1%K2HPO4、0.2%吐温80、1 mmol/L Fe2+,初始pH为自然值(7.0).优化后的摇瓶发酵条件是:36℃、转速180 r/min、500 mL三角瓶中加入50 mL培养基、接种量2%.采用优化后的培养基发酵培养18 h酶活力达到最大值10.71 XU/mL.与优化前相比,产酶最高峰提前6h,而木聚糖酶活力是优化前的2.5倍,本实验得到的优化发酵条件大大提高了重组菌产酶能力.

  10. The conditions of the liquid fermentation of pleurotus ostreatus to produce xylanase using brewer's spent grain as raw materials%利用啤酒糟液体深层培养风尾菇产木聚糖酶的初步研究

    Institute of Scientific and Technical Information of China (English)

    潘真清; 陈涛

    2010-01-01

    研究了风尾菇(Pleurotus ostreatus)利用啤酒糟作为原料进行液态发酵产木聚糖酶的可行性.通过正交试验得出最佳的培养基配方.同时对不同发酵时间还原糖的减少和木聚糖酶活性的进行测定.培养基最佳配方:啤酒糟6 g,黄豆粉0.32 g,玉米粉2 g,糖10 g,pH值5.0.发酵的最佳时间为72 h,此时木聚糖酶的活性最大,为9.2 U/mL.

  11. Purification and Properties of 43 kD Xylanase XYNA from Streptomyces olivaceoviridis A1%橄榄绿链霉菌A1所产43kD木聚糖酶XYNA的纯化及其酶学性质

    Institute of Scientific and Technical Information of China (English)

    张红莲; 姚斌; 袁铁铮; 王亚茹; 操时树; 范云六

    2002-01-01

    橄榄绿链霉菌A1(Streptomyces olivaceoviridisA1)所产木聚糖酶XYNA经阴离子交换层析和分子筛层析分离,得到纯化的XYNA.XYNA的分子量约为43 kD.其最适温度为60℃,最适pH5.6.XYNA在55℃下以4-O-Me-D-glucurono-D-xylan的Km值和Vmax分别是332 g/kg和15.72μmol/(mL@min).SDS、EDTA对XYNA有轻微的抑制作用.XYNA无纤维素酶活性.胃蛋白酶、胰蛋白酶处理30 min,对酶活性无影响.XYNA成熟蛋白N端10个氨基酸序列为Ala-G1u-Ser-Thr-Leu-Gly-Ala-A1a-Ala-Ala.

  12. 一株嗜热菌产耐热木聚糖酶对馒头品质和保质期的影响%Effect of Thermo-tolerant Xylanase from Thermophilic Geobacillus sp.PZH1 on the Quality and Shelf-life of Steamed Bread

    Institute of Scientific and Technical Information of China (English)

    王石峰; 林孔亮; 秦晓培; 陈学敏; 刘培培; 郭小虎; 张波

    2011-01-01

    研究嗜热细菌Geobacillus sp.PZH1产耐热木聚糖酶对馒头品质及保质期的影响作用.从嗜热细菌Geobacillus sp.的PZH1制备木聚糖酶,添加到面粉中制作馒头,观察其对馒头品质和保质期的影响.结果表明:在馒头中添加适量的耐热木聚糖酶,能明显降低馒头的持水性,提高面筋网络的弹性,改变面团的加工及稳定性能,增大馒头体积,并能有效抑制馒头中细菌的生长,延长馒头的保质期.

  13. Influence of Xylan Content on the Oxygen Delignification Performance of Eucalypt Kraft Pulps as Studied Using Prehydrolysis and Xylanase Treatment%聚木糖含量对硫酸盐桉木浆氧脱木素性能的影响

    Institute of Scientific and Technical Information of China (English)

    马倩倩

    2014-01-01

    评价氧脱木素效率的常用参数有卡伯值和Klason木素含量.聚木糖含量的变化通常会导致己烯糖醛酸(HexA)含量的变化,用卡伯值衡量氧脱木素程度时应考虑HexA的影响.由于硫酸盐阔叶木浆中残余木素量较少,因此以Klason木素含量法评价氧脱木素效率的精确性值得商榷.通过用聚木糖酶处理未漂硫酸盐浆和用酸预水解硫酸盐法制浆,获得了聚木糖含量不同的纸浆,探讨了聚木糖含量对硫酸盐巨尾桉浆氧脱木素效率的影响.实验结果表明,随着聚木糖的去除,以HexA修正的卡伯值表示的氧脱木素程度无显著变化,以Klason木素含量表示的卡伯值的变化趋势也不明显.

  14. Content of Pentosan in Wheat Grain and Xylanase Supplementation on Measurements of AME and Nutrient Ingredient Digestibility of Cocks%小麦戊聚糖含量及添加木聚糖酶对鸡表观代谢能值和养分消化率的影响

    Institute of Scientific and Technical Information of China (English)

    王修启; 李春喜; 林东康; 常绢; 秦磊

    2002-01-01

    测定了河南省9个主栽小麦品种豫麦49号、豫麦47号、高优503、郑麦9023、豫麦34号、皖麦38、豫麦70号、孟12和河北8901的戊聚糖含量,结果表明,9个品种的戊聚糖含量为6.25%~8.23%;选取两个有代表性的品种豫麦70号和豫麦49号,用64只小公鸡进行代谢试验,评定添加1.2‰木聚糖酶对鸡表观代谢能(AME)值和养分消化率的影响.

  15. The Construction of A Metagenomic Library of Uncultured Bacteria from Compost and the Cloning and Identification of Novel Xylanase Genes%堆肥未培养细菌的宏基因组文库构建及新的木聚糖酶基因的克隆和鉴定

    Institute of Scientific and Technical Information of China (English)

    张鹏; 段承杰; 庞浩; 封毅; 靳振江; 许跃强; 莫新春; 唐纪良; 冯家勋

    2005-01-01

    从自制堆肥样品中提取未培养细菌的总DNA,用柯斯质粒pWEB∷TNC为载体构建宏基因组文库,对文库进行筛选获得表达木聚糖酶活性的克隆,再进行亚克隆、测序分析以及Blastx搜索GenBank分析木聚糖酶基因.结果构建得到一个包含约5万个克隆的宏基因组文库,文库中外源DNA总容量约为1.8×106 kb,获得2个表达木聚糖酶活性的克隆:pGXN1050和pGXN1051,鉴定分析表明:pGXN1050上潜在的木聚糖酶基因umxyn11A具1个771bp的ORF(Open Reading Frame),可编码含257个氨基酸的蛋白质,所编码产物与混合纤维弧菌(Cellvibrio mixtus)的内切-1,4-β-木聚糖酶(GenBank索引号Z48925.1)的氨基酸序列有46%的一致性和57%的相似性;pGXN1051上潜在的木聚糖酶基因umxyn11B具一个723bp的ORF,可编码含241个氨基酸的蛋白质,编码产物与混合纤维弧菌的内切-1,4-β-木聚糖酶(GenBank索引号Z48925.1)的氨基酸序列有73%的一致性和80%的相似性.木聚糖酶Umxyn11A和Umxyn11B都属于糖基水解酶家族11的成员.

  16. Study on the optimum medium component of liquid fermentation by xylanase and its enzyme characteristics%一株霉菌产木聚糖酶液体发酵培养基组分优化及酶学性质的研究

    Institute of Scientific and Technical Information of China (English)

    吴萍; 饶圣宏; 马忠友; 张生

    2008-01-01

    本试验主要对一株产木聚糖酶B45菌株的液体发酵培养基组分进行优化.通过单因素和正交试验确定了所试因素的最佳组合,并对其酶学性质进行研究.结果表明:B45菌株液体发酵培养基中的最优组分为:碳源(玉米芯)2%,氮源(玉米粉)0.2%,无机盐(KH22PO4+MgSO4)0.1%,微量元素(FeSO4)0.0015%.木聚糖酶的最适反应pH为5.0,最适反应温度为50℃,有较好的热稳定性,但不耐强酸(pH≥4)和碱性(pH≤7).

  17. 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.

  18. HMMCAS: a web tool for the identification and domain annotations of Cas proteins.

    Science.gov (United States)

    Chai, Guoshi; Yu, Min; Jiang, Lixu; Duan, Yaocong; Huang, Jian

    2017-02-07

    The CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated proteins) adaptive immune systems are discovered in many bacteria and most archaea. These systems are encoded by cas (CRISPR-associated) operons that have an extremely diverse architecture. The most crucial step in the depiction of cas operons composition is the identification of cas genes or Cas proteins. With the continuous increase of the newly sequenced archaeal and bacterial genomes, the recognition of new Cas proteins is becoming possible, which not only provides candidates for novel genome editing tools but also helps to understand the prokaryotic immune system better. Here we describe HMMCAS, a web service for the detection of CRISPR-associated structural and functional domains in protein sequences. HMMCAS uses hmmscan similarity search algorithm in HMMER3.1 to provide a fast, interactive service based on a comprehensive collection of hidden Markov models of Cas protein family. It can accurately identify the Cas proteins including those fusion proteins, for example the Cas1-Cas4 fusion protein in Candidatus Chloracidobacterium thermophilum B (Cab. thermophilum B). HMMCAS can also find putative cas operon and determine which type it belongs to. HMMCAS is freely available at http://i.uestc.edu.cn/hmmcas.

  19. Ancestral genetic diversity associated with the rapid spread of stress-tolerant coral symbionts in response to Holocene climate change

    KAUST Repository

    Hume, Benjamin C. C.

    2016-04-05

    Coral communities in the Persian/Arabian Gulf (PAG) withstand unusually high salinity levels and regular summer temperature maxima of up to ∼35 °C that kill conspecifics elsewhere. Due to the recent formation of the PAG and its subsequent shift to a hot climate, these corals have had only <6, 000 y to adapt to these extreme conditions and can therefore inform on how coral reefs may respond to global warming. One key to coral survival in the world\\'s warmest reefs are symbioses with a newly discovered alga, Symbiodinium thermophilum. Currently, it is unknown whether this symbiont originated elsewhere or emerged from unexpectedly fast evolution catalyzed by the extreme environment. Analyzing genetic diversity of symbiotic algae across >5, 000 km of the PAG, the Gulf of Oman, and the Red Sea coastline, we show that S. thermophilum is a member of a highly diverse, ancient group of symbionts cryptically distributed outside the PAG. We argue that the adjustment to temperature extremes by PAG corals was facilitated by the positive selection of preadapted symbionts. Our findings suggest that maintaining the largest possible pool of potentially stress-tolerant genotypes by protecting existing biodiversity is crucial to promote rapid adaptation to present-day climate change, not only for coral reefs, but for ecosystems in general.

  20. Effect of amount of straw provided to growing/finishing pigs on gastric ulceration at slaughter

    DEFF Research Database (Denmark)

    Herskin, Mette S; Jensen, Henrik E.; Jespersen, A.;

    2014-01-01

    The effect of a current commercial xylanase (DAN) and experimental xylanase (EX), and EX in combination with protease (EXP), on the degradation and apparent ileal digestibility (AID) of non-starch polysaccharides (NSP) in wheat Distillers Dried Grains with Solubles (DDGS), was studied in 8 ileum-...... enzymes increase the degradation of NSP and arabinoxylan in wheat DDGS, with ranking of DAN > EXP > EX. Keywords: DDGS, non-starch polysaccharide digestibility, xylanase......The effect of a current commercial xylanase (DAN) and experimental xylanase (EX), and EX in combination with protease (EXP), on the degradation and apparent ileal digestibility (AID) of non-starch polysaccharides (NSP) in wheat Distillers Dried Grains with Solubles (DDGS), was studied in 8 ileum......-cannulated pigs (initial BW 36.6±2.8 kg) following a double 4x4 Latin Square design. The control and three enzyme diets, each containing 96% DDGS, were supplemented with vitamins, minerals, L-lysine, 500 FTU phytase/kg feed, dust-binder and chromic oxide (3 g/kg). The pigs were fed 3 times daily for 1 week...

  1. Detection of Extracellular Enzyme Activity in Penicillium using Chromogenic Media.

    Science.gov (United States)

    Yoon, Ji Hwan; Hong, Seung Beom; Ko, Seung Ju; Kim, Seong Hwan

    2007-09-01

    A total of 106 Penicillium species were tested to examine their ability of degrading cellobiose, pectin and xylan. The activity of β-glucosidase was generally strong in all the Penicillium species tested. P. citrinum, P. charlesii, P. manginii and P. aurantiacum showed the higher ability of producing β-glucosidase than other tested species. Pectinase activity was detected in 24 Penicillium species. P. paracanescens, P. sizovae, P. sartoryi, P. chrysogenum, and P. claviforme showed strong pectinase activity. In xylanase assay, 84 Penicillium species showed activity. Strong xylanase activity was detected from P. megasporum, P. sartoryi, P. chrysogenum, P. glandicola, P. discolor, and P. coprophilum. Overall, most of the Penicillium species tested showed strong β-glucosidase activity. The degree of pectinase and xylanase activity varied depending on Penicillium species.

  2. Increasing efficiency of enzymatic hemicellulose removal from bamboo for production of high-grade dissolving pulp.

    Science.gov (United States)

    Zhao, Lingfeng; Yuan, Zhaoyang; Kapu, Nuwan Sella; Chang, Xue Feng; Beatson, Rodger; Trajano, Heather L; Martinez, D Mark

    2017-01-01

    To improve the efficiency of enzymatic hemicellulose removal from bamboo pre-hydrolysis kraft pulp, mechanical refining was conducted prior to enzyme treatment. Refining significantly improved the subsequent hemicellulose removal efficiency by xylanase treatment. Results showed that when PFI refining was followed by 3h xylanase treatment, the xylan content of the bamboo pre-hydrolysis kraft pulp (after first stage oxygen delignification) could be decreased to 2.72% (w/w). After bleaching of enzyme treated pulp, the alpha-cellulose content was 93.4% (w/w) while the xylan content was only 2.38%. The effect of refining on fibre properties was investigated in terms of freeness, water retention value, fibre length and fibrillation characteristics. The brightness, reactivity and viscosity were also determined to characterize the quality of final pulp. Results demonstrated the feasibility of combining refining and xylanase treatment to produce high quality bamboo dissolving pulp.

  3. Hydrolyzabilities of different corn stover fractions after aqueous ammonia pretreatment.

    Science.gov (United States)

    Sun, Zongping; Ge, Xiaoyan; Xin, Donglin; Zhang, Junhua

    2014-02-01

    The effect of aqueous ammonia pretreatment on the hydrolysis of different corn stover fractions (rind, husk, leaf, and pith) by xylanase (XYL) with cellulases (CELs) was evaluated. The aqueous ammonia pretreatment had excellent delignification ability (above 66%) for different corn stover fractions. The corn rind exhibited the lowest susceptibility to aqueous ammonia pretreatment. The pretreated rind showed the lowest hydrolyzability by CEL and XYL, which was supported by a high content of crystalline cellulose in the hydrolyzed residues of rind, as confirmed by X-ray diffraction (XRD). With the addition of 1 mg XYL/g dry matter, a high glucose yield (above 90%) could be obtained from the pretreated rind by CEL. The results revealed that a high hydrolyzate yield of corn rind after aqueous ammonia pretreatment could be obtained with 1 mg xylanase/g dry matter, showing that aqueous ammonia pretreatment and xylanase addition to cellulases have great potential for the efficient hydrolysis of corn stover without previous fractionation.

  4. 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;

    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 fungal symbiont. CONCLUSION...

  5. DETERMINATION OF ENZYMES PRODUCED BY CERIPORIOPSIS SUBVERMISPORA DURING PRETREATMENT OF DIFFERENT BIOMASS SOURCES

    Directory of Open Access Journals (Sweden)

    Miroslav Ondrejovič

    2012-02-01

    Full Text Available The aim of this paper was to study of lignocellulolytic enzymes producing by Ceriporiopsis subvermispora during its cultivation on three types of plant biomass differentiated by chemical composition and physical properties (wheat straw, pine and poplar wood. The activity of lignocellulolytic enzymes in cultivation medium was determined by catalytic transformation of their natural substrates to products which were detected by photometric methods. Cellulase activities were very low while xylanases predominated. Wheat straw was best substrate for production of cellulases (4.38 U/mL and xylanases (23.34 U/mL. The maximum activity of cellulase and xylanase was reached at 8th and 3rd day, respectively. Laccase activity reached the maximum after 16 days and then gradually decreased. The best substrate for production of laccases was poplar wood (1.67 U/mL.

  6. 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...... 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...

  7. Effect of selected natural dyes in reduction on colour changes of Egyptian linen textiles by fungi.

    Science.gov (United States)

    Abdel-Kareem, Omar

    2007-07-01

    Linen is the most historical Egyptian textile fibre liable to fungal deterioration. Fungal deterioration of dyed linen textiles may appear as undesirable different stains. In order to success in removing of fungal stains from biodeteriorated historical Egyptian dyed linen textiles, it is necessary to understand the nature and causes of these stains, hence their subsequent removal. So this paper aims to investigate the effect of fungi on dyed linen textiles. In this study linen textile samples were experimentally dyed by two different dyes, blue one as an example to vat dye and yellow one as an example to direct dye. This work is done on two of the most important dyes (Turmeric and indigo), which were popular in most of historical periods in Egypt. Dyed linen samples were experimentally biodegraded by thirty different fungal strains isolated previously from historical Egyptian linen samples. The produced change in colours of the biodeteriorated samples was detected visually. Also, the change in reflection spectra and colour differences produced to dyed linen textiles after fungal deterioration, were assessed and evaluated by using spectrophotometer. This study reported that most of tested fungi contribute to discoloration of all tested dyed linen samples. These results indicate that most of stains on historical Egyptian dyed linen textiles, may be fungal stains. The results confirm that undyed linen textiles more liable to fungal biodeterioration than dyed ones. Also the results show that yellow dyed linen textiles are more susceptible to fungal deterioration than blue dyed linen textiles. The obtained results show that Alternaria tenuissima, Chaetomium globosum, Chaetomium sp., Penicillium raistrickii, P. soppi, P. asperum, P. citrinum, Aspergillus carbonarius, A. fischeri, A. nidulans, A. terreus and A. niger, had showed the maximum colour changes of the deteriorated yellow dyed linen samples. The results also show that Alternaria tenuissima, Chaetomium sp

  8. 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

  9. 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.

  10. Structural insights into the specificity of Xyn10B from Paenibacillus barcinonensis and its improved stability by forced protein evolution.

    Science.gov (United States)

    Gallardo, Oscar; Pastor, F I Javier; Polaina, Julio; Diaz, Pilar; Łysek, Robert; Vogel, Pierre; Isorna, Pablo; González, Beatriz; Sanz-Aparicio, Julia

    2010-01-22

    Paenibacillus barcinonensis is a soil bacterium bearing a complex set of enzymes for xylan degradation, including several secreted enzymes and Xyn10B, one of the few intracellular xylanases reported to date. The crystal structure of Xyn10B has been determined by x-ray analysis. The enzyme folds into the typical (beta/alpha)(8) barrel of family 10 glycosyl hydrolases (GH10), with additional secondary structure elements within the beta/alpha motifs. One of these loops -L7- located at the beta7 C terminus, was essential for xylanase activity as its partial deletion yielded an inactive enzyme. The loop contains residues His(249)-Glu(250), which shape a pocket opened to solvent in close proximity to the +2 subsite, which has not been described in other GH10 enzymes. This wide cavity at the +2 subsite, where methyl-2,4-pentanediol from the crystallization medium was found, is a noteworthy feature of Xyn10B, as compared with the narrow crevice described for other GH10 xylanases. Docking analysis showed that this open cavity can accommodate glucuronic acid decorations of xylo-oligosaccharides. Co-crystallization experiments with conduramine derivative inhibitors supported the importance of this open cavity at the +2 subsite for Xyn10B activity. Several mutant derivatives of Xyn10B with improved thermal stability were obtained by forced evolution. Among them, mutant xylanases S15L and M93V showed increased half-life, whereas the double mutant S15L/M93V exhibited a further increase in stability, showing a 20-fold higher heat resistance than the wild type xylanase. All the mutations obtained were located on the surface of Xyn10B. Replacement of a Ser by a Leu residue in mutant xylanase S15L can increase hydrophobic packing efficiency and fill a superficial indentation of the protein, giving rise to a more compact structure of the enzyme.

  11. Why are Aspergilli so different in their expression of secondary metabolites from section to section?

    DEFF Research Database (Denmark)

    Frisvad, Jens Christian; Rank, Christian; Larsen, Thomas Ostenfeld

    for example Bipolaris, Chaetomium, Humicola, & Podospora. On the other hand SMs such as the ochratoxins have only been found in Aspergillus and Penicillium so far, and the aflatoxins have only been found in Aspergillus. Within Aspergillus, which comprises 9 very different teleomorphs (Eurotium, Chaetosartorya...... produced by species in different sections (Fig. 1, Fig. 2). Aspergillic acids and ochratoxins have been found in Flavi and Circumdati, pseurotins have been found in Fumigati and Clavati , kojic acid + aflatoxins have been found in few Nidulantes but many Flavi species; cyclopiazonic acid has been found...

  12. Biological Control of Olive Green Mold in Agaricus bisporus Cultivation.

    Science.gov (United States)

    Tautorus, T E; Townsley, P M

    1983-02-01

    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 not only provided a significant degree of protection from C. olivaceum, but also increased yields of Agaricus bisporus.

  13. The occurrence of keratinophilic fungi in sewage sludge from Egypt.

    Science.gov (United States)

    Abdel-Hafez, A I; el-Sharouny, H M

    1990-01-01

    The keratinophilic fungi of 40 sewage sludge samples from Upper Egypt were studied using a goat hair-baiting technique. 43 species representing 22 genera were isolated, 17 species of which were dermatophytes and closely related fungi: Chrysosporium state of Arthroderma tuberculatum, C. asperatum, C. georgii, C. indicum, C. keratinophilum, C. pseudomerdarium, C. queenslandicum, Chrysosporium state of Thielavia sepedonium, C. tropicum, Microsporum cookei, M. gypseum, Myceliophthora anamorph of Corynascus novoguineensis, M. vellerea and Trichophyton terrestre. 26 species of cycloheximide resistant fungi were collected and these included members of Acremonium, Aspergillus, Alternaria, Chaetomium, Cladosporium, Cunninghamella, Emericella, Fusarium, Geotrichum, Penicillium and others.

  14. 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.

  15. 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.

  16. Distribution frequency and incidence of seed-borne pathogens of some cereals and industrial crops in Serbia

    OpenAIRE

    Jelena Lević; Slavica Stankоvić; Vesna Krnjaja; Aleksandra Bočarov-Stančić; Dragica Ivanović

    2012-01-01

    A total of 41 species of fungi were isolated from seed samples of barley, maize, soybean, and sunflower collected at different locations in Serbia. The majority of detected species occurred on barley (35 of 41 species or 87.8%) comparing to soybean (17 of 41 species or 41.5%), sunflower (16 of 41 species or 39.0%) and maize (15 of 41 species or 36.9%). Species belonging to genera Alternaria, Chaetomium, Epicoccum, Fusarium, Penicillium and Rhizopus were present on seeds of all four plant spec...

  17. Degradation of toxaphene by Bjerkandera sp. strain BOL13 using waste biomass as a cosubstrate.

    Science.gov (United States)

    Lacayo Romero, Martha; Terrazas, Enrique; van Bavel, Bert; Mattiasson, Bo

    2006-07-01

    The white-rot fungus Bjerkandera sp. strain BOL13 was capable of degrading toxaphene when supplied with wood chips, wheat husk or cane molasses as cosubstrates in batch culture experiments. Approximately 85% of toxaphene was removed when wheat husk was the main substrate. The production of lignin peroxidase was only stimulated when wheat husk was present in the liquid medium. Although xylanase was always detected, wheat husk supported the highest xylanase production. A negligible amount of beta-glucosidase and cellulase were found in the batch culture medium. To the best of our knowledge, this is the first reported case of toxaphene degradation by white-rot fungi.

  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. Examining the Potential of Plasma-Assisted Pretreated Wheat Straw for Enzyme Production by Trichoderma reesei

    DEFF Research Database (Denmark)

    Rodríguez Gómez, Divanery; Lehmann, Linda Olkjær; Schultz-Jensen, Nadja

    2012-01-01

    Plasma-assisted pretreated wheat straw was investigated for cellulase and xylanase production by Trichoderma reesei fermentation. Fermentations were conducted with media containing washed and unwashed plasma-assisted pretreated wheat straw as carbon source which was sterilized by autoclavation....... To account for any effects of autoclavation, a comparison was made with unsterilized media containing antibiotics. It was found that unsterilized washed plasma-assisted pretreated wheat straw (which contained antibiotics) was best suited for the production of xylanases (110 IU ml(-1)) and cellulases (0...

  20. 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

    -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....

  1. Hydrogen production by anaerobic microbial communities exposed to repeated heat treatments.

    Science.gov (United States)

    Duangmanee, T; Padmasiri, S I; Simmons, J J; Raskin, L; Sung, S

    2007-09-01

    Biological hydrogen production by anaerobic mixed communities was studied in laboratory-scale bioreactors using sucrose as the substrate. A bioreactor in which a fraction of the return sludge was exposed to repeated heat treatments performed better than a control bioreactor without repeated heat treatment of return sludge and produced a yield of 2.15 moles of hydrogen per mole of sucrose, with 50% hydrogen in the biogas. Terminal restriction fragment length polymorphism analysis showed that two different Clostridium groups (comprised of one or more species) were dominant during hydrogen production. The relative abundance of two other non-Clostridium groups increased during periods of decreased hydrogen production. The first group consisted of Bifidobacterium thermophilum, and the second group included one or more of Bacillus, Melissococcus, Spirochaeta, and Spiroplasma spp.

  2. Intramolecular cross-linking in a bacterial homolog of mammalian SLC6 neurotransmitter transporters suggests an evolutionary conserved role of transmembrane segments 7 and 8

    DEFF Research Database (Denmark)

    Kniazeff, Julie; Loland, Claus Juul; Goldberg, Naomi;

    2005-01-01

    The extracellular concentration of the neurotransmitters dopamine, serotonin, norepinephrine, GABA and glycine is tightly controlled by plasma membrane transporters belonging to the SLC6 gene family. A very large number of putative transport proteins with a remarkable homology to the SLC6...... transporters has recently been identified in prokaryotes. Here we have probed structural relationships in a 'microdoman' corresponding to the extracellular ends of transmembrane segments (TM) 7 and 8 in one of these homologs, the tryptophan transporter TnaT from Symbiobacterium thermophilum. We found...... proximity between TM 7 and 8 in the tertiary structure of TnaT as previously suggested for the mammalian counterparts. Furthermore, the inhibition of uptake upon cross-linking the two cysteines provides indirect support for a conserved conformational role of these transmembrane domains in the transport...

  3. Dicty_cDB: Contig-U09796-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 1) RecName: Full=Nuclease sbcCD subunit C; &AP008934_1411... 36 1.2 ( O76329 ) RecName: Full=Interaptin; Alt...epticum strain ... 33 7.6 (Q2YXX0) RecName: Full=Nuclease sbcCD subunit C; 33 7.6... ( P44928 ) RecName: Full=Multidrug resistance protein A homolog; ... 33 10.0 (Q8NWV1) RecName: Full=Nuclease sbc...01393 |pid:none) Anaerocellum thermophilum DSM 6... 33 10.0 (A5ISM9) RecName: Full=Nuclease sbcCD subunit C;...FYT3) RecName: Full=Nuclease sbcCD subunit C; 33 10.0 CP001107_490( CP001107 |pid

  4. 一幅霉变书画的扫描电镜分析%Scanning electron microscopic analysis of a mold-contaminated painting

    Institute of Scientific and Technical Information of China (English)

    杨娟

    2015-01-01

    利用扫描电镜分析一幅霉变清代书画样品上菌落的表面结构和形貌,明确了污染书画上的霉菌为毛壳菌。通过对纸张纤维超微结构分析显示:毛壳菌对纸张纤维结构的破坏作用不容忽视。此研究为后期文物的修复和保存提供了超微形态学依据。%The ultra⁃morphology of fungi on the contaminated sample from a Qing dynasty painting was examined by using digital microscope and scanning electron microscope. The fungi belongs to Chaetomium species. The analysis of paper fiber structure suggests that Chaetomium species have devastating effect on paper which cannot be overlooked in any cases. The results provide ultramicroscopic morphology evidence for future conservation and restoration of paintings.

  5. 山东海岸木生海洋真菌的研究I.%WOOD-INHABITING MARINE FUNGI FROM THE COAST OF SHANDONG I.

    Institute of Scientific and Technical Information of China (English)

    金静; 李桂舫; 李保华; 张天宇; 梁晨; 李宝笃

    2004-01-01

    为调查中国黄海海域的丝状海洋真菌,从山东威海潮间带海滩收集了沉没木、附着木和沙埋木,并从其上分离到7种高等海洋真菌.Arenariomyces trifurcata,Corollospora maritima,Alternaria maritima,Trichocladium achrasporum为中国大陆新记录种,Chaetomium gl0b0sum,Tetraploa aristita,Torula sp.为中国大陆新生境报道.对每个种作了描述并对分类和形态进行了讨论.%To survey the filamentous marine fungi from Yellow Sea of China, submerged wood, trap wood amongst rocks, and sand-buried wood were taken from the intertidal beach sites in Weihai,Shandong Province. Seven species of higher marine fungi were found. Arenariomyces trifurcata,Corollospora maritima, Alternaria maritima and Trichocladium achrasporum were new records to mainland of China; Chaetomium globosum, Tetraploa aristita and Torula sp. were found for the first time in the sea report in mailand of China. Taxonomy and morphology of these species were discussed.

  6. 海南菠萝几种叶部真菌病害研究%Study on Several Fungal Leaf Diseases of Pineapple of Hainan

    Institute of Scientific and Technical Information of China (English)

    罗志文; 范鸿雁; 李向宏; 刘银叶; 华敏; 王祥和; 余乃通; 周文静; 何凡

    2012-01-01

    对海南5个菠萝产区菠萝叶部真菌病害进行了调查研究.结果表明,共有6种真菌病害发生较普遍,危害较重,分别由胶孢炭疽菌Colletotrichum gloeosporioides、画眉草弯孢菌Curvularia eragrostidis、刺环裂壳孢菌Annellolacinia dinemasporioides、掌状拟盘多毛孢菌Pestalotiopsis palmarum、喙状凸脐蠕孢菌Exserohilum rostratum和金色毛壳菌Chaetomium aureum引起,其中喙状凸脐蠕孢菌 E.rostratum和金色毛壳菌C.aureum为我国首次报道.本文同时对6种病害的发病症状和病原形态进行了初步描述.%The fungal diseases on pineapple (Ananas comosus (L. ) Merr. ) leaf were primarily investigated in 5 producing regions of Hainan. The result showed that 6 kinds of fungal leaf diseases, caused by Colletotrichum gloeosporioides, Cwvularia eragrostidis, Annellolacinia dinemasporioidcs, Pestalotiopsis palmarum, Exxrohilum rostratum and Chaetomium aureum, respectively, were found to widely occur and had severe harm here, and the E. rostratum and C. aureum were firstly reported on pineapple in China. Besides, the symptoms and pathogen morphology of 6 kinds of fungal leaf diseases were primarily described.

  7. Main: LECPLEACS2 [PLACE

    Lifescience Database Archive (English)

    Full Text Available LECPLEACS2 S000465 24-April-2005 (last modified) kehi Core element in LeCp (tomato ...Cys protease) binding cis-element (from -715 to -675) in LeAcs2 gene; cysteine protease; ethylene; xylanase; ACS; Lycopersicon esculentum (tomato) TAAAATAT ...

  8. Monosaccharides and Ethanol Production from Superfine Ground Sugarcane Bagasse Using Enzyme Cocktail

    Directory of Open Access Journals (Sweden)

    Jingbo Li

    2014-03-01

    Full Text Available In this work, the effect of particle size on the enzymatic hydrolysis of milled and sieved sugarcane bagasse (SCB was studied. The enzymatic hydrolysis and fermentability of superfine ground SCB (SGP400 using an enzyme cocktail strategy were also explored. Particle size reduction improved the enzymatic hydrolysis. The highest glucose yield was 44.75%, which was obtained from SGP400. The enzyme cocktail strategy greatly enhanced the glucose and xylose yield. The maximum glucose and xylose yield was from the enzyme cocktail of cellulase, xylanase, and pectinase. Synergistic action between xylanase and pectinase as well as cellulase and pectinase was quite noticeable. Hydrolysis times affected the degree of synergism. Ethanol production was carried out by employing simultaneous saccharification and fermentation (SSF and semi-SSF using enzymes and their cocktails. Semi-SSF was found to be the better one compared with SSF. Xylanase and pectinase aided the ethanol production in both fermentation modes. Ethanol yield was 7.81 and 7.30 g/L for semi-SSF and SSF, respectively by using an enzyme cocktail of cellulase, β-glucosidase, pectinase, and xylanase.

  9. Metagenomic analysis of novel lignocellulose-degrading enzymes from higher termite guts inhabiting microbes.

    Science.gov (United States)

    Nimchua, Thidarat; Thongaram, Taksawan; Uengwetwanit, Tanaporn; Pongpattanakitshote, Somchai; Eurwilaichitr, Lily

    2012-04-01

    A metagenomic fosmid library was constructed from genomic DNA isolated from the microbial community residing in hindguts of a wood-feeding higher termite (Microcerotermes sp.) collected in Thailand. The library was screened for clones expressing lignocellulolytic activities. Fourteen independent active clones (2 cellulases and 12 xylanases) were obtained by functional screening at pH 10.0. Analysis of shotgun-cloning and pyrosequencing data revealed six ORFs, which shared less than 59% identity and 73% similarity of their amino acid sequences with known cellulases and xylanases. Conserved domain analysis of these ORFs revealed a cellulase belonging to the glycoside hydrolase family 5, whereas the other five xylanases showed significant identity to diverse families including families 8, 10, and 11. Interestingly, one fosmid clone was isolated carrying three contiguous xylanase genes that may comprise a xylanosome operon. The enzymes with the highest activities at alkaline pH from the initial activity screening were characterized biochemically. These enzymes showed a broad range of enzyme activities from pH 5.0 to 10.0, with pH optimal of 8.0 retaining more than 70% of their respective activities at pH 9.0. The optimal temperatures of these enzymes ranged from 50 degrees C to 55 degrees C. This study provides evidence for the diversity and function of lignocellulose-degrading enzymes in the termite gut microbial community, which could be of potential use for industrial processes such as pulp biobleaching and denim biostoning.

  10. Main: 1TA3 [RPSD[Archive

    Lifescience Database Archive (English)

    Full Text Available 1TA3 小麦 Bread Wheat Triticum aestivum Xylanase Inhibitor Protein I Precursor. Name=...479.1; -.|PDB; 1OM0; X-ray; A=31-304.|PDB; 1TA3; X-ray; A=-.|PDB; 1TE1; X-ray; A=...CGYPPAAHVGRALATGIFERAHVRTYESDKWCNQNLGWEGSWDKWTAAYPATRFYVGLTADDKSHQWVHPKNVYYGVAPVAQKKDNYGGIMLWDRYFDKQTNYSSLIKYYA wheat_1TA3.jpg ...

  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 ( nutritional value of wheat middlings-based diets.

  12. 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...

  13. 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.

  14. Supplementing enzymes to extruded, soybean based diet improves breakdown of non-starch polysaccharides in rainbow trout (Oncorhynchus mykiss)

    DEFF Research Database (Denmark)

    Dalsgaard, Anne Johanne Tang; Knudsen, Knud Erik Bach; Verlhac, Viviane

    2016-01-01

    presumably by assisting in the breakdown of NSP. This study examined the effects on NSP degradation when supplementing β-glucanase, xylanase, protease or a mix of the three enzymes to an extruded, juvenile rainbow trout (Oncorhynchus mykiss) diet containing 344 g kg−1 de-hulled, solvent-extracted soybean...

  15. The global regulator LaeA controls production of citric acid and endoglucanases in Aspergillus carbonarius

    DEFF Research Database (Denmark)

    Linde, Tore; Zoglowek, Marta; Lübeck, Mette;

    2016-01-01

    The global regulatory protein LaeA is known for regulating the production of many kinds of secondary metabolites in Aspergillus species, as well as sexual and asexual reproduction, and morphology. In Aspergillus carbonarius, it has been shown that LaeA regulates production of ochratoxin. We have ...... production or on cellulose or xylanase activity....

  16. Hyperthermostable cellulolytic and hemicellulolytic enzymes and their biotechnological applications

    OpenAIRE

    Tipparat Hongpattarakere

    2002-01-01

    Hyperthermal cellulases and hemicellulases have been intensively studied due to their highly potential applications at extreme temperatures, which mimic industrial processes involving cellulose and hemicellulose degradation. More than 50 species of hyperthermophiles have been isolated, many of which possess hyperthermal enzymes required for hydrolyzing cellulose and hemicelluloses. Endoglucanases, exoglucanases, cellobiohydrolases, xylanases, β-glucosidase and β-galactosidase, which are produ...

  17. Optimization of hydrothermal pretreatment of wheat straw for production of bioethanol at low water consumption without addition of chemicals

    DEFF Research Database (Denmark)

    Østergaard Petersen, Mai; Larsen, Jan; Thomsen, Mette Hedegaard

    2009-01-01

    In the IBUS process (Integrated Biomass Utilization System) lignocellulosic biomass is converted into ethanol at high dry matter content without addition of chemicals and with a strong focus on energy efficiency. This study describes optimization of continuous hydrothermal pretreatment of wheat s...... cellulase mixtures - increasing to 92% when adding a commercial xylanase. (C) 2009 Elsevier Ltd. All rights reserved....

  18. 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

  19. Search for cell-wall-degrading enzymes of world-wide rice grains by PCR and their effects on the palatability of rice.

    Science.gov (United States)

    Nakamura, Sumiko; Machida, Keisuke; Ohtsubo, Ken'ichi

    2012-01-01

    Such rice cultivars as Japonica, Japonica-Indica hybrid, Javanica and Indica, were evaluated for their main chemical components (amylose content and protein content), pasting property of rice flour (consistency), physical property of the cooked rice grains (adhesion, L3), and enzyme activities (cellulase and xylanase). The amylose content, cellulase activity and xylanase activity showed significant positive or negative correlation with the pasting property (consistency) of rice flour (r = 0.89, r = 0.58, r = 0.70, respectively) and with the physical property of the cooked rice grains (adhesion, L3: r = -0.51, r = -0.61, r = -0.71, respectively) at the level of 1%. Endogenous xylanase and cellulase played important roles to determine the texture of the cooked rice grains similarly to the amylose content. Part of the DNA sequences of the α-glucosidase gene differed among the Japonica, Japonica-Indica hybrid and Indica subspecies. We found discriminative DNA bands appearing by PCR, corresponding to 1,4-β-xylanase and endo-1,4-β-glucanase 13 in the case of Indica rice, Indica-Japonica hybrid rice, and Javanica rice (non-Japonica subspecies). The equation for estimating the physical property (adhesion) of cooked rice grains by PCR was improved by adding novel primers related to the cell-wall-degrading enzymes.

  20. 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