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Sample records for fungal enzyme preparations

  1. Safety evaluation of a fungal pectinesterase enzyme preparation and its use in food.

    Science.gov (United States)

    Lissau, B G; Pedersen, P B; Petersen, B R; Budolfsen, G

    1998-01-01

    The Aspergillus aculeatus pectinesterase enzyme is used to modify the texture of plant derived products. It is produced by A. oryzae transformed with the cloned full length cDNA of A. aculeatus encoding pectinesterase. It was subjected to a series of toxicological tests to document safety in use. The enzyme preparation was not found to be mutagenic in the Ames test, and did not cause chromosomal damage in a human lymphocyte assay. In a 13-week oral-toxicity study in rats, with daily dosages up to 10 g enzyme preparation kg body weight (b.w.), there were no adverse effects on mortality, clinical signs, body weight, food or water consumption, ophthalmoscopic findings, haematology or clinical chemistry. There were also no notable necropsy or histological findings. Statistically significant increases in heart weight were noted in male animals treated with 5 or 10 g enzyme preparation/kg b.w./day, following covariance analysis. However, this was not considered to be related to treatment with the enzyme preparation. The issue of the levels of free liberation of methanol in products processed with pectinesterase is addressed, and it is concluded that, from a nutritional and physiological point of view, free as well as bound methanol must be considered.

  2. Irradiation of industrial enzyme preparations. II. Characterization of fungal pectinase by thin-layer isoelectric focusing and gel filtration

    Energy Technology Data Exchange (ETDEWEB)

    Delincee, H.

    1978-01-01

    Industrial dry fungal pectinase from A. niger was irradiated with doses (up to 1 Mrad) of /sup 60/Co-..gamma..rays effective in reducing microbial contamination. The pectinase was characterized by thin-layer isoelectric focusing and gel filtration in order to detect possible radiation-induced structural alterations. Thin-layer isoelectric focusing revealed at least fifteen multiple forms with pectin-depolymerizing activity, with isoelectric points in the range pH 4.5 to 7. Heterogeneity of pectinesterase was also demonstrated, the main band occurring around pH 4. By thin-layer gel filtration the molecular weight of the pectin-depolymerase was estimated as being about 36,000, and that of pectinesterase as about 33,000. Radiation-induced changes of the charge properties or molecular size of the irradiated pectinase preparation were not observed. The feasibility of using ionizing radiation for the reduction of microbial contamination of industrial enzyme preparations looks promising.

  3. Polyphosphate present in DNA preparations from fungal species of Collectotrichum inhibits restriction endonucleases and other enzymes

    Science.gov (United States)

    Rodriguez, R.J.

    1993-01-01

    During the development of a procedure for the isolation of total genomic DNA from filamentous fungi (Rodriguez, R. J., and Yoder, 0. C., Exp. Mycol. 15, 232-242, 1991) a cell fraction was isolated which inhibited the digestion of DNA by restriction enzymes. After elimination of DNA, RNA, proteins, and lipids, the active compound was purified by gel filtration to yield a single fraction capable of complete inhibition of restriction enzyme activity. The inhibitor did not absorb uv light above 220 nm, and was resistant to alkali and acid at 25°C and to temperatures as high as 100°C. More extensive analyses demonstrated that the inhibitor was also capable of inhibiting T4 DNA ligase and TaqI DNA polymerase, but not DNase or RNase. Chemical analyses indicated that the inhibitor was devoid of carbohydrates, proteins, lipids, and nucleic acids but rich in phosphorus. A combination of nuclear magnetic resonance, metachromatic shift of toluidine blue, and gel filtration indicated that the inhibitor was a polyphosphate (polyP) containing approximately 60 phosphate molecules. The mechanism of inhibition appeared to involve complexing of polyP to the enzymatic proteins. All species of Colletotrichum analyzed produced polyP equivalent in chain length and concentration. A modification to the original DNA extraction procedure is described which eliminates polyP and reduces the time necessary to obtain DNA of sufficient purity for restriction enzyme digestion and TaqI polymerase amplification.

  4. Hydrolysis of solubilized hemicellulose derived from wet-oxidized wheat straw by a mixture of commercial fungal enzyme preparations

    Energy Technology Data Exchange (ETDEWEB)

    Skammelsen Schmidt, Anette; Thomsen, Alle Belinda; Woidemann, Anders [Risoe National Lab. (Denmark); Tenkanen, Maija [VTT Biotechnology and Food Research (Finland)

    1998-04-01

    The enzymatic hydrolysis of the solubilized hemicellulose fraction from wet-oxidized wheat straw was investigated for quantification purposes. An optimal hydrolysis depends on factors such as composition of the applied enzyme mixture and the hydrolysis conditions (enzyme loading, hydrolysis time, pH-value, and temperature). A concentrated enzyme mixture was used in this study prepared at VTT Biotechnology and Food Research, Finland, by mixing four commercial enzyme preparations. No distinctive pH-value and temperature optima were identified after a prolonged incubation of 24 hours. By reducing the hydrolysis time to 2 hours a temperature optimum was found at 50 deg. C, where a pH-value higher than 5.2 resulted in reduced activity. An enzyme-substrate-volume-ratio of 0.042, a pH-value of 5.0, and a temperature of 50 deg. C were chosen as the best hydrolysis conditions due to an improved monosaccharide yield. The hydrolysis time was chosen to be 24 hours to ensure equilibrium and total quantification. Even under the best hydrolysis conditions, the overall sugar yield from the enzymatic hydrolysis was only 85% of that of the optimal acid hydrolysis. The glucose yield were approximately the same for the two types of hydrolyses, probably due to the high cellulase activity in the VTT-enzyme mixture. For xylose and arabinose the enzymatic hydrolysis yielded only 80% of that of the acid hydrolysis. As the pentoses existed mainly as complex polymers their degradation required many different enzymes, some of which might be missing from the VTT-enzyme mixture. Furthermore, the removal of side-choins from the xylan backbone during the wet-oxidation pretreatment process might enable the hemicellulosic polymers to interact and precipitate, hence, reducing the enzymatic digestibility of the hemicellulose. (au) 8 tabs., 10 ills., 65 refs.

  5. Modelling Fungal Fermentations for Enzyme Production

    DEFF Research Database (Denmark)

    Albæk, Mads Orla; Gernaey, Krist; Hansen, Morten S.

    We have developed a process model of fungal fed-batch fermentations for enzyme production. In these processes, oxygen transfer rate is limiting and controls the substrate feeding rate. The model has been shown to describe cultivations of both Aspergillus oryzae and Trichoderma reesei strains in 550...

  6. Fungal enzymes in the attine ant symbiosis

    DEFF Research Database (Denmark)

    de Fine Licht, Henrik Hjarvard; Schiøtt, Morten; Boomsma, Jacobus Jan

    build huge nests displacing several cubic meters of soil, whereas lower attine genera such as Cyphomyrmex and Mycocepurus have small nests with a fungus garden the size of a table-tennis ball. Only the leaf-cutter ants are specialized on using fresh leaves as substrate for their fungus gardens, whereas...... or different efficiencies of enzyme function. Fungal enzymes that degrade plant cell walls may have functionally co-evolved with the ants in this scenario. We explore this hypothesis with direct measurements of enzyme activity in fungus gardens in 12 species across 8 genera spanning the entire phylogeny...

  7. Chitinases biosynthesis by immobilized Aeromonas hydrophila SBK1 by prawn shells valorization and application of enzyme cocktail for fungal protoplast preparation.

    Science.gov (United States)

    Halder, Suman Kumar; Maity, Chiranjit; Jana, Arijit; Ghosh, Kuntal; Das, Arpan; Paul, Tanmay; Das Mohapatra, Pradeep Kumar; Pati, Bikas Ranjan; Mondal, Keshab Chandra

    2014-02-01

    Production and optimization of β-N-acetyl glucosaminidase and chitinase by Ca-alginate immobilized Aeromonas hydrophila SBK1 was carried out using prawn shell as cost-effective substrate. Beads prepared with 5.0% Na-alginate (containing 2.0% colloidal chitin) and 1.0 M CaCl2 showed considerable beads integrity and supported maximum production of chitinolytic enzymes. Bead diameter, 3 mm; temperature, 35°C; pH 7.0; agitation, 90 rpm were found ideal for the maximum production of the enzymes. The fermentation and thermodynamic indices revealed the feasibility of immobilized cells over free cells for enzymes production. Reasonable amount of chitosaccharides (degree of polymerization; 1-6) accumulated in the production media which have paramount antioxidant activity. Scale up experiment was successfully carried out in 5 L fermentor. In immobilized state, the chitosaccharides yield and antioxidant activity increased about 44.76% and 22.22%, whereas specific productivity of β-N-acetyl glucosaminidase and chitinase increased by 22.86% and 33.37% over free state. The cell entrapped beads can be reused upto ten cycles without marked loss of its biocatalytic efficiency. High level of protoplast of Aspergillus niger was generated by treating mycelia with 10 U/ml of crude chitinase after 4 h at pH 7.0 and in the temperature 35-40°C, and 67% of the protoplasts were found to be regenerated. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  8. Evolution of Fungal enzymes in the attine ant symbiosis

    DEFF Research Database (Denmark)

    de Fine Licht, Henrik Hjarvard; Schiøtt, Morten; Boomsma, Jacobus Jan

    The attine ant symbiosis is characterized by ancient but varying degrees of diffuse co-evolution between the ants and their fungal cultivars. Domesticated fungi became dependent on vertical transmission by queens and the ant colonies came to rely on their symbiotic fungus for food and thus......, indirectly, on fungal enzymes to break down the plant material brought in by the ants as fungal substrate. The more than 210 extant fungus-growing ant species differ considerably in colony size, social complexity and substrate-use. Only the derived leaf-cutting ants are specialized on using fresh leaves...... as garden substrate, whereas the more basal genera use leaf litter, insect feces and insect carcasses. We hypothesized that enzyme activity of fungal symbionts has co-evolved with substrate use and we measured enzyme activities of fungus gardens in the field to test this, focusing particularly on plant...

  9. Production of amylase enzyme from mangrove fungal isolates ...

    African Journals Online (AJOL)

    The mangrove ecosystem serves as a bioresource for various industrially important microorganisms. The use of fungi as a source of industrially relevant enzymes led to an increased interest in the application of microbial enzymes in various industrial processes. Fungal colonies were isolated from sediments of five different ...

  10. Evolution of Fungal enzymes in the attine ant symbiosis

    DEFF Research Database (Denmark)

    de Fine Licht, Henrik Hjarvard; Schiøtt, Morten; Boomsma, Jacobus Jan

    The attine ant symbiosis is characterized by ancient but varying degrees of diffuse co-evolution between the ants and their fungal cultivars. Domesticated fungi became dependent on vertical transmission by queens and the ant colonies came to rely on their symbiotic fungus for food and thus...... as garden substrate, whereas the more basal genera use leaf litter, insect feces and insect carcasses. We hypothesized that enzyme activity of fungal symbionts has co-evolved with substrate use and we measured enzyme activities of fungus gardens in the field to test this, focusing particularly on plant...... essential for the symbiosis in general, but have contributed specifically to the evolution of the symbiosis....

  11. Production of amylase enzyme from mangrove fungal isolates

    African Journals Online (AJOL)

    sunny

    2014-11-12

    Nov 12, 2014 ... Production of amylase enzyme from mangrove ... Production medium. The production medium used for growth of the fungal isolate was soluble starch 50 g, yeast extract 0.5 g, KH2PO4 10 g, (NH4)2SO4 ..... Very good (Upper part- Light brownish, lower- yellowish black, Pigment- yellow to black soluble). MA.

  12. 21 CFR 864.4400 - Enzyme preparations.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Enzyme preparations. 864.4400 Section 864.4400...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Specimen Preparation Reagents § 864.4400 Enzyme preparations. (a) Identification. Enzyme preparations are products that are used in the histopathology...

  13. Production of theabrownins using a crude fungal enzyme concentrate.

    Science.gov (United States)

    Wang, Qiuping; Gong, Jiashun; Chisti, Yusuf; Sirisansaneeyakul, Sarote

    2016-08-10

    Theabrownins were produced from infusions of sun-dried green tea leaves using a crude enzyme concentrate of Aspergillus tubingensis TISTR 3647. This fungus had been isolated from a solid state fermentation of Pu-erh type tea. The crude enzyme concentrate contained activities of peroxidase, catechol oxidase and laccase. The enzyme concentrate effectively oxidized the phenolic compounds in green tea infusion to theabrownins. A theabrownins concentration of 56.0g/L was obtained in 44h. The reaction mixture contained the green tea infusion and crude enzyme concentrate in the volume ratio of 1: 0.205. The tea infusion had been produced using 200g of tea leaves per liter of distilled water. The reaction was carried out in a stirred bioreactor at 37°C with an aeration rate of 1 vvm, an agitation speed of 250rpm and a controlled pH of 7.0. Peroxidase, catechol oxidase, and laccase acted synergistically to convert the phenolic compounds in green tea infusion to theabrownins. Previously, theabrownins had been produced from green tea infusions only by using live fungal cultures. Production using the microorganism-free enzyme concentrate was comparable to production using the fungus A. tubingensis TISTR 3647. The proposed novel production process using the fungal crude enzymes and green tea infusion, offers a more controlled, reproducible and highly productive option for commercial production of theabrownins. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Bacterial and Fungal Proteolytic Enzymes: Production, Catalysis and Potential Applications.

    Science.gov (United States)

    da Silva, Ronivaldo Rodrigues

    2017-09-01

    Submerged and solid-state bioprocesses have been extensively explored worldwide and employed in a number of important studies dealing with microbial cultivation for the production of enzymes. The development of these production technologies has facilitated the generation of new enzyme-based products with applications in pharmaceuticals, food, bioactive peptides, and basic research studies, among others. The applicability of microorganisms in biotechnology is potentiated because of their various advantages, including large-scale production, short time of cultivation, and ease of handling. Currently, several studies are being conducted to search for new microbial peptidases with peculiar biochemical properties for industrial applications. Bioprospecting, being an important prerequisite for research and biotechnological development, is based on exploring the microbial diversity for enzyme production. Limited information is available on the production of specific proteolytic enzymes from bacterial and fungal species, especially on the subgroups threonine and glutamic peptidases, and the seventh catalytic type, nonhydrolytic asparagine peptide lyase. This gap in information motivated the present study about these unique biocatalysts. In this study, the biochemical and biotechnological aspects of the seven catalytic types of proteolytic enzymes, namely aspartyl, cysteine, serine, metallo, glutamic, and threonine peptidase, and asparagine peptide lyase, are summarized, with an emphasis on new studies, production, catalysis, and application of these enzymes.

  15. Fungal Enzymes and Yeasts for Conversion of Plant Biomass to Bioenergy and High-Value Products.

    Science.gov (United States)

    Lange, Lene

    2017-01-01

    Fungi and fungal enzymes play important roles in the new bioeconomy. Enzymes from filamentous fungi can unlock the potential of recalcitrant lignocellulose structures of plant cell walls as a new resource, and fungi such as yeast can produce bioethanol from the sugars released after enzyme treatment. Such processes reflect inherent characteristics of the fungal way of life, namely, that fungi as heterotrophic organisms must break down complex carbon structures of organic materials to satisfy their need for carbon and nitrogen for growth and reproduction. This chapter describes major steps in the conversion of plant biomass to value-added products. These products provide a basis for substituting fossil-derived fuels, chemicals, and materials, as well as unlocking the biomass potential of the agricultural harvest to yield more food and feed. This article focuses on the mycological basis for the fungal contribution to biorefinery processes, which are instrumental for improved resource efficiency and central to the new bioeconomy. Which types of processes, inherent to fungal physiology and activities in nature, are exploited in the new industrial processes? Which families of the fungal kingdom and which types of fungal habitats and ecological specializations are hot spots for fungal biomass conversion? How can the best fungal enzymes be found and optimized for industrial use? How can they be produced most efficiently-in fungal expression hosts? How have industrial biotechnology and biomass conversion research contributed to mycology and environmental research? Future perspectives and approaches are listed, highlighting the importance of fungi in development of the bioeconomy.

  16. 21 CFR 184.1372 - Insoluble glucose isomerase enzyme preparations.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Insoluble glucose isomerase enzyme preparations... enzyme preparations. (a) Insoluble glucose isomerase enzyme preparations are used in the production of... additional requirements for enzyme preparations in the Food Chemicals Codex, 3d Ed. (1981), p. 107, which is...

  17. Bacterial and fungal keratitis in Upper Egypt: In vitro screening of enzymes, toxins and antifungal activity

    Directory of Open Access Journals (Sweden)

    Abdullah A Gharamah

    2014-01-01

    Full Text Available Purpose: This work was conducted to study the ability of bacterial and fungal isolates from keratitis cases in Upper Egypt to produce enzymes, toxins, and to test the isolated fungal species sensitivity to some therapeutic agents. Materials and Methods: One hundred and fifteen patients clinically diagnosed to have microbial keratitis were investigated. From these cases, 37 bacterial isolates and 25 fungal isolates were screened for their ability to produce extra-cellular enzymes in solid media. In addition, the ability of fungal isolates to produce mycotoxins and their sensitivity to 4 antifungal agents were tested. Results: Protease, lipase, hemolysins, urease, phosphatase, and catalase were detected respectively in 48.65%, 37.84%, 59.46%, 43.24%, 67.57%, and 100% out of 37 bacterial isolates tested. Out of 25 fungal isolates tested during the present study, 80% were positive for protease, 84% for lipase and urease, 28% for blood hemolysis, and 100% for phosphatase and catalase enzymes. Thirteen fungal isolates were able to produce detectable amounts of 7 mycotoxins in culture medium (aflatoxins (B1, B2, G1, and G2, sterigmatocystin, fumagillin, diacetoxyscirpenol, zearalenone, T-2 toxin, and trichodermin. Among the antifungal agents tested in this study, terbinafine showed the highest effect against most isolates in vitro. Conclusion: In conclusion, the ability of bacterial and fungal isolates to produce extracellular enzymes and toxins may be aid in the invasion and destruction of eye tissues, which, in turn, lead to vision loss.

  18. Immunoassays of fungal laccases for screening of natural enzymes and control of recombinant enzyme production.

    Science.gov (United States)

    Loginov, Dmitry S; Vavilova, Ekaterina A; Savinova, Оlga S; Abyanova, Alfia R; Chulkin, Andrey M; Vasina, Daria V; Zherdev, Anatoly V; Koroleva, Olga V

    2014-01-01

    Because of the wide application of laccases in different biotechnological processes and intense studies of the enzymes from different sources, the development of efficient techniques for monitoring laccase level is a task of significant importance. Enzyme-linked immunosorbent assay (ELISA) and Western blotting techniques were developed to control total content and isoform composition of laccases, including their recombinant preparations. Because glycosylated and nonglycosylated forms have different structures and sets of epitopes, two kinds of polyclonal antibodies were obtained and applied. The first antibody recognized the native (glycosylated) laccase purified from Trametes hirsuta and the second one reacted with recombinant (nonglycosylated) laccase expressed in Escherichia coli. Titers of the antibodies were analyzed by indirect ELISA with laccases isolated from several strains of basidiomycetes. The obtained cross-reactivity data for both antibodies demonstrated a correspondence with sequence homology of the laccases. The antibodies raised against recombinant (nonglycosylated) laccase had higher titers and thus were preferable for screening of recombinant laccase in cultural media. Thus, optimal antibody preparations were selected for screening of laccase-producing strains, and the control of recombinant enzymes and the efficiency of their use in immunochemical control of laccase levels were confirmed. © 2013 International Union of Biochemistry and Molecular Biology, Inc.

  19. Enzymes and fungal virulence | Tonukari | Journal of Applied ...

    African Journals Online (AJOL)

    This paper presents a comprehensive literature review of cell wall degrading enzymes (CWDEs). Plant pathogenic fungi secrete extracellular enzymes that are capable of degrading the cell walls of their host plants. These CWDEs may be necessary for penetration of the cell wall barrier, as well as for generation of simple ...

  20. Fungal and enzyme treatment kidney: a promising way to help pulp and paper mills to achieve zero effluent discharge

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, X.; Soong, J.J.; Stebbing, D.W.; Saddler, J.N. [British Columbia Univ., Vancouver, BC (Canada). Forest Products Biotechnology Faculty of Forestry; Beatson, R.P. [British Columbia Inst. of Technology, Burnaby, BC (Canada). Advanced Papermaking Initiative

    2001-06-01

    Water usage and effluent discharge must be minimized by pulp and paper mills to comply with increasingly stringent environmental standards and market demands. As a result, the potential use of a combined fungal and enzyme system was evaluated as an internal water treatment kidney in a TMP/newsprint mill with a closed water system. The evaluation involved the use of three different white water samples. A sample of mill white water (MWW) was taken from the cloudy white water chest of the disk thickener at the Howe Sound Pulp and Paper Ltd. A mix of interior spruce/pine/fir and coastal hemlock from British Columbia comprised the chip supply used during the collection. Then , the model recycled white water (RWW) was prepared, as was the membrane filtered model recycled white water (FWW). A significant decrease in the amount of total dissolved and colloidal substances was observed as a result of the growth of the white-rot fungus Trametes versicolor on these waters. After seven days of fungal treatment, in excess of 75 per cent of the extractives, and 62-71 per cent of the carbohydrates initially present in the three white waters were removed. Laccase, cellulase and lipase enzyme activities had been noted in the fungal culture filtrates (FCF) during the growth of the fungus. Using the fungal culture filtrates obtained after two days growth of Trametes versicolor, subsequent fungal enzyme treatments of MWW and RWW were performed on mill white water. After a three-hour FCF treatment at 65 Celsius, more than 90 per cent of the lignans and ester bonded extractives (steryl esters and triglycerides) were removed from both white waters. During the same period, the resin and fatty acids content decreased by almost 40 per cent in the mill white water while it decreased by almost 60 per cent in the model white water sample. The polymerization of low molecular weight phenolics into higher molecular weight lignin-types of material occurred as a result of the fungal and enzyme

  1. Lytic Polysaccharide Monooxygenases - Studies of Fungal Secretomes and Enzyme Properties

    DEFF Research Database (Denmark)

    Nekiunaite, Laura

    degradation, were also identified upstream the LPMO genes, providing evidence for a co-regulatory mechanism of LPMOs and amylolytic hydrolases. The second part of the PhD thesis is focused on understanding the binding properties of LPMOs to starch and starch mimic substrate. It was shown that LPMOs possessing...... to different substrates at the protein level. It could help to design better enzyme cocktails that increase efficiency of biomass degradation. The secretomes of A. nidulans revealed differences in growth and secretion of enzymes, depending on the type and properties of starches. A common characteristic...... conversion as they produce a wide diversity of degrading enzymes. In the first part of this PhD thesis, the secretomes of the well-known fungus Aspergillus nidulans grown on cereal and legume starches were analyzed. Secretomics is a powerful tool to unravel secretion patterns of fungi and their response...

  2. The prominent role of fungi and fungal enzymes in the ant-fungus biomass conversion symbiosis.

    Science.gov (United States)

    Lange, L; Grell, M N

    2014-06-01

    Molecular studies have added significantly to understanding of the role of fungi and fungal enzymes in the efficient biomass conversion, which takes place in the fungus garden of leaf-cutting ants. It is now clear that the fungal symbiont expresses the full spectrum of genes for degrading cellulose and other plant cell wall polysaccharides. Since the start of the genomics era, numerous interesting studies have especially focused on evolutionary, molecular, and organismal aspects of the biological and biochemical functions of the symbiosis between leaf-cutting ants (Atta spp. and Acromyrmex spp.) and their fungal symbiont Leucoagaricus gongylophorus. Macroscopic observations of the fungus-farming ant colony inherently depict the ants as the leading part of the symbiosis (the myrmicocentric approach, overshadowing the mycocentric aspects). However, at the molecular level, it is fungal enzymes that enable the ants to access the nutrition embedded in recalcitrant plant biomass. Our hypothesis is that the evolutionary events that established fungus-farming practice were predisposed by a fascinating fungal evolution toward increasing attractiveness to ants. This resulted in the ants allowing the fungus to grow in the nests and began to supply plant materials for more fungal growth. Molecular studies also confirm that specialized fungal structures, the gongylidia, with high levels of proteins and rich blend of enzymes, are essential for symbiosis. Harvested and used as ant feed, the gongylidia are the key factor for sustaining the highly complex leaf-cutting ant colony. This microbial upgrade of fresh leaves to protein-enriched animal feed can serve as inspiration for modern biorefinery technology.

  3. Screening for cellulose and hemicellulose degrading enzymes from the fungal genus Ulocladium

    DEFF Research Database (Denmark)

    Pedersen, Mads; Hollensted, Morten; Lange, L.

    2009-01-01

    The fungal genus Ulocladium consists mostly of saprotrophic species and can readily be isolated from dead vegetation, rotten wood. paper, textiles and other cellulose containing materials. Thus, they must produce cellulolytic and hemicellulolytic enzymes. In this study fifty Ulocladium strains fr...

  4. Fungal Morphology in Industrial Enzyme Production - Modelling and Monitoring

    DEFF Research Database (Denmark)

    Quintanilla, D.; Hagemann, T.; Hansen, K.

    2015-01-01

    Filamentous fungi are widely used in the biotechnology industry for the production of industrial enzymes. Thus, considerable work has been done with the purpose of characterizing these processes. The ultimate goal of these efforts is to be able to control and predict fermentation performance......, and on the way the data is interpreted-i.e. which models were applied. The main filamentous fungi used in industrial fermentation are introduced, ranging from Trichoderma reesei to Aspergillus species. Due to the fact that secondary metabolites, like antibiotics, are not to be considered bulk products, organisms...... that spans across strains and scales, as most studies indeed conclude that their respective results might be strain specific and not necessarily valid across scales....

  5. Evaluation of a Hypocrea jecorina Enzyme Preparation for Hydrolysis of Tifton 85 Bermudagrass

    Science.gov (United States)

    Ximenes, E. A.; Brandon, S. K.; Doran-Peterson, J.

    Tifton 85 bermudagrass, developed at the ARS-USDA in Tifton, GA, is grown on over ten million acres in the USA for hay and forage. Of the bermudagrass cultivars, Tifton 85 exhibits improved digestibility because the ratio of ether- to ester-linked phenolic acids has been lowered using traditional plant breeding techniques. A previously developed pressurized batch hot water (PBHW) method was used to treat Tifton 85 bermudagrass for enzymatic hydrolysis. Native grass (untreated) and PBHW-pretreated material were compared as substrates for fungal cultivation to produce enzymes. Cellulase activity, measured via the filter paper assay, was higher for fungi cultivated on PBHW-pretreated grass, whereas the other nine enzyme assays produced higher activities for the untreated grass. Ferulic acid and vanillin levels increased significantly for the enzyme preparations produced using PBHW-pretreated grass and the release of these phenolic compounds may have contributed to the observed reduction in enzyme activities. Culture supernatant from Tifton 85 bermudagrass-grown fungi were combined with two commercial enzyme preparations and the enzyme activity profiles are reported. The amount of reducing sugar liberated by the enzyme mixture from Hypocrea jecorina (after 192 h incubation with untreated bermudagrass) individually or in combination with feruloyl esterase was 72.1 and 84.8%, respectively, of the commercial cellulase preparation analyzed under the same conditions.

  6. Development of a freeze-dried fungal wettable powder preparation able to biodegrade chlorpyrifos on vegetables.

    Directory of Open Access Journals (Sweden)

    Jie Liu

    Full Text Available Continuous use of the pesticide chlorpyrifos has resulted in harmful contaminations in environment and species. Based on a chlorpyrifos-degrading fungus Cladosporium cladosporioides strain Hu-01 (collection number: CCTCC M 20711, a fungal wettable powder preparation was developed aiming to efficiently remove chlorpyrifos residues from vegetables. The formula was determined to be 11.0% of carboxymethyl cellulose-Na, 9.0% of polyethylene glycol 6000, 5.0% of primary alcohol ethoxylate, 2.5% of glycine, 5.0% of fucose, 27.5% of kaolin and 40% of freeze dried fungi by response surface methodology (RSM. The results of quality inspection indicated that the fungal preparation could reach manufacturing standards. Finally, the degradation of chlorpyrifos by this fungal preparation was determined on pre-harvest cabbage. Compared to the controls without fungal preparation, the degradation of chlorpyrifos on cabbages, which was sprayed with the fungal preparation, was up to 91% after 7 d. These results suggested this freeze-dried fungal wettable powder may possess potential for biodegradation of chlorpyrifos residues on vegetables and provide a potential strategy for food and environment safety against pesticide residues.

  7. Fungal Community and Ligninolytic Enzyme Activities in Quercus deserticola Trel. Litter from Forest Fragments with Increasing Levels of Disturbance

    Directory of Open Access Journals (Sweden)

    Jesús A. Rosales-Castillo

    2017-12-01

    Full Text Available Litter fungal communities and their ligninolytic enzyme activities (laccase, Mn-peroxidase, and lignin-peroxidase play a vital role in forest biogeochemical cycles by breaking down plant cell wall polymers, including recalcitrant lignin. However, litter fungal communities and ligninolytic enzyme activities have rarely been studied in Neotropical, non-coniferous forests. Here, we found no significant differences in litter ligninolytic enzyme activities from well preserved, moderately disturbed, and heavily disturbed Quercus deserticola Trel. forests in central Mexico. However, we did find seasonal effects on enzyme activities: during the dry season, we observed lower laccase, and increased Mn-peroxidase and lignin-peroxidase activities, and in the rainy season, Mn-peroxidase and lignin-peroxidase activities were lower, while laccase activity peaked. Fungal diversity (Shannon-Weaver and Simpson indices based on ITS-rDNA analyses decreased with increased disturbance, and principal component analysis showed that litter fungal communities are structured differently between forest types. White-rot Polyporales and Auriculariales only occurred in the well preserved forest, and a high number of Ascomycota were shared between forests. While the degree of forest disturbance significantly affected the litter fungal community structure, the ligninolytic enzyme activities remained unaffected, suggesting functional redundancy and a possible role of generalist Ascomycota taxa in litter delignification. Forest conservation and restoration strategies must account for leaf litter and its associated fungal community.

  8. Method of preparing cross-linked enzyme particles

    NARCIS (Netherlands)

    Mateo, C.; Van Langen, L.M.; Van Rantwijk, F.

    2004-01-01

    The invention relates to a method of preparing cross-linked enzyme particles using a cross-linking agent. According to the invention, the enzyme particles are formed and subsequently cross-linked using a cross-linking agent having at least n reactive groups where N>=3 and a molecular weight of

  9. Variation in fungal enzyme spectra may affect mutualistic division of labour between ants and fungus gardens

    DEFF Research Database (Denmark)

    de Fine Licht, Henrik Hjarvard; Boomsma, Jacobus Jan

    partners vary in metabolic performance, division of labour may not always be optimized and co-evolutionary trajectories become less predictable. The higher fungus-growing (attine) ants consist of the leafcutter ants (Acromyrmex and Atta), which rear a single fungal species throughout their Latin American...... in garden enzyme activity independent of ant species association. This illustrates that the mosaic of coevolutionary interactions in Trachymyrmex and Sericomyrmex fungus-growers may be fundamentally different from the interactions in Acromyrmex and, particularly, Atta leaf-cutting ants. The former appear...

  10. Fungal inhibitors of proteolytic enzymes: classification, properties, possible biological roles, and perspectives for practical use.

    Science.gov (United States)

    Dunaevsky, Y E; Popova, V V; Semenova, T A; Beliakova, G A; Belozersky, M A

    2014-06-01

    Peptidase inhibitors are ubiquitous regulatory proteins controlling catalytic activity of proteolytic enzymes. Interest in these proteins increased substantially after it became clear that they can be used for therapy of various important diseases including cancer, malaria, and autoimmune and neurodegenerative diseases. In this review we summarize available data on peptidase inhibitors from fungi, emphasizing their properties, biological role, and possible practical applications of these proteins in the future. A number of fungal peptidase inhibitors with unique structure and specificity of action have no sequence homology with other classes of peptidase inhibitors, thus representing new and specific candidates for therapeutic use. The main classifications of inhibitors in current use are considered. Available data on structure, mechanisms and conditions of action, and diversity of functions of peptidase inhibitors of fungi are analyzed. It is mentioned that on one side the unique properties of some inhibitors can be used for selective inhibition of peptidases responsible for initiation and development of pathogenic processes. On the other side, general inhibitory activity of other inhibitors towards peptidases of various catalytic classes might be able to provide efficient defense of transgenic plants against insect pests by overcoming compensatory synthesis of new peptidases by these pests in response to introduction of a fungal inhibitor. Together, the data analyzed in this review reveal that fungal inhibitors extend the spectrum of known peptidase inhibitors potentially suitable for use in medicine and agriculture. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  11. Isolation and Screening of Thermo-Stable Cellulase Enzyme Fungal Producer at Different Temperature

    International Nuclear Information System (INIS)

    Noor Ashiqin Jamroo; Noor Azrimi Umor; Kamsani

    2015-01-01

    Thermo stable cellulase from fungi has high potential for industrial application. In this study, wild -type of fungal were isolate from different sources such as hot spring water, sea water, soft wood, rice straw and cow dung. The isolates were characterized by cultural and morphological observation. Based on morphological characteristics, the genera of all fungal cultures were identified namely Aspergillus fumigatus. The screening for thermo stable cellulase were done using 2 % carboxymethyl cellulose and congo red as an indicator at temperature 30, 37, 45 and 50 degree Celsius respectively. Out of 26 fungal isolates, only eight isolates were selected for further screening and showed the abilities to secrete cellulases by forming distinct halo zones on selective agar plate. The maximum halo zone ranging from 32 mm to 35 mm were obtained after 72 hour incubation at 50 degree Celsius by H2, SW1 and C1 isolates. As contrary other isolates showed halo zone range from 22 mm to 29 mm at same temperature. All the isolates showed the abilities to secrete cellulase enzyme at other temperature but lower when compared to 50 degree Celsius referred to the halo zone obtained. The SW1 isolates showed highest cellulolytic index which was 2.93 measured at 37 degree Celsius and 2.67 at 50 degree Celsius respectively. (author)

  12. Multifunctional Cellulolytic Enzymes Outperform Processive Fungal Cellulases for Coproduction of Nanocellulose and Biofuels.

    Science.gov (United States)

    Yarbrough, John M; Zhang, Ruoran; Mittal, Ashutosh; Vander Wall, Todd; Bomble, Yannick J; Decker, Stephen R; Himmel, Michael E; Ciesielski, Peter N

    2017-03-28

    Producing fuels, chemicals, and materials from renewable resources to meet societal demands remains an important step in the transition to a sustainable, clean energy economy. The use of cellulolytic enzymes for the production of nanocellulose enables the coproduction of sugars for biofuels production in a format that is largely compatible with the process design employed by modern lignocellulosic (second generation) biorefineries. However, yields of enzymatically produced nanocellulose are typically much lower than those achieved by mineral acid production methods. In this study, we compare the capacity for coproduction of nanocellulose and fermentable sugars using two vastly different cellulase systems: the classical "free enzyme" system of the saprophytic fungus, Trichoderma reesei (T. reesei) and the complexed, multifunctional enzymes produced by the hot springs resident, Caldicellulosiruptor bescii (C. bescii). We demonstrate by comparative digestions that the C. bescii system outperforms the fungal enzyme system in terms of total cellulose conversion, sugar production, and nanocellulose production. In addition, we show by multimodal imaging and dynamic light scattering that the nanocellulose produced by the C. bescii cellulase system is substantially more uniform than that produced by the T. reesei system. These disparities in the yields and characteristics of the nanocellulose produced by these disparate systems can be attributed to the dramatic differences in the mechanisms of action of the dominant enzymes in each system.

  13. Fungal Enzymes for Bio-Products from Sustainable and Waste Biomass.

    Science.gov (United States)

    Gupta, Vijai K; Kubicek, Christian P; Berrin, Jean-Guy; Wilson, David W; Couturier, Marie; Berlin, Alex; Filho, Edivaldo X F; Ezeji, Thaddeus

    2016-07-01

    Lignocellulose, the most abundant renewable carbon source on earth, is the logical candidate to replace fossil carbon as the major biofuel raw material. Nevertheless, the technologies needed to convert lignocellulose into soluble products that can then be utilized by the chemical or fuel industries face several challenges. Enzymatic hydrolysis is of major importance, and we review the progress made in fungal enzyme technology over the past few years with major emphasis on (i) the enzymes needed for the conversion of polysaccharides (cellulose and hemicellulose) into soluble products, (ii) the potential uses of lignin degradation products, and (iii) current progress and bottlenecks for the use of the soluble lignocellulose derivatives in emerging biorefineries. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Cellulose digestion in Monochamus marmorator Kby. (coleoptera: Cerambycidae): role of acquired fungal enzymes

    International Nuclear Information System (INIS)

    Kukol, J.J.; Martin, M.M.

    1986-01-01

    Larvae of the balsam fir sawyer, Monochamus marmorator Kby. (Coleoptera, Cerambycidae), contain midgut digestive enzymes active against hemicellulose and cellulose. Cellulases from larvae fed on balsam fir wood infected with the fungus, Trichoderma harzianum Rifai (Deuteromycetes, Moniliales, Moniliaceae), were found to be identical to those of the cellulase complex produced by this fungus when compared using chromatography, electrophoresis, and isofocusing. When larvae are maintained on a fungusfree diet, their midgut fluids lack cellulolytic activity, and they are unable to digest cellulose. Cellulolytic capacity can be restored by feeding the larvae wood permeated by fungi. We conclude that the enzymes which enable M. marmorator larvae to digest cellulose are not produced by the larvae. Instead, the larvae acquire the capacity to digest cellulose by ingesting active fungal cellulases while feeding in fungus-infected wood

  15. Cellulose digestion in Monochamus marmorator Kby. (coleoptera: Cerambycidae): role of acquired fungal enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Kukol, J.J.; Martin, M.M.

    1986-05-01

    Larvae of the balsam fir sawyer, Monochamus marmorator Kby. (Coleoptera, Cerambycidae), contain midgut digestive enzymes active against hemicellulose and cellulose. Cellulases from larvae fed on balsam fir wood infected with the fungus, Trichoderma harzianum Rifai (Deuteromycetes, Moniliales, Moniliaceae), were found to be identical to those of the cellulase complex produced by this fungus when compared using chromatography, electrophoresis, and isofocusing. When larvae are maintained on a fungusfree diet, their midgut fluids lack cellulolytic activity, and they are unable to digest cellulose. Cellulolytic capacity can be restored by feeding the larvae wood permeated by fungi. We conclude that the enzymes which enable M. marmorator larvae to digest cellulose are not produced by the larvae. Instead, the larvae acquire the capacity to digest cellulose by ingesting active fungal cellulases while feeding in fungus-infected wood.

  16. Continuous enzyme reactions with immobilized enzyme tubes prepared by radiation cast-polymerization

    International Nuclear Information System (INIS)

    Kumakura, Minoru; Kaetsu, Isao

    1986-01-01

    Immobilized glucose oxidase tubes were prepared by radiation cast-polymerization of 2-hydroxyethyl methacrylate and tetraethyleneglycol diacrylate monomer at low temperatures. The immobilized enzyme tubes which were spirally set in a water bath were used as reactor, in which the enzyme activity varied with tube size and flow rate of the substrate. The conversion yield of the substrate in continuous enzyme reaction was about 80%. (author)

  17. Synthetic inhibitors of the fungal detoxifying enzyme brassinin oxidase based on the phytoalexin camalexin scaffold.

    Science.gov (United States)

    Pedras, M Soledade C; Minic, Zoran; Sarma-Mamillapalle, Vijay K

    2009-03-25

    Brassinin (1) is an essential phytoalexin produced in plants of the family Brassicaceae (common name crucifer) due to its role as a biosynthetic precursor of other phytoalexins and antimicrobial activity. The dithiocarbamate group of brassinin (1) is the toxophore responsible for its fairly broad antifungal activity. To the detriment of many agriculturally important crops, several pathogenic fungi of crucifers are able to overcome brassinin by detoxification. In this work, inhibitors of brassinin oxidase, a phytoalexin detoxifying enzyme produced by the plant pathogenic fungus Leptosphaeria maculans (asexual stage Phoma lingam ), were synthesized and evaluated. The camalexin scaffold was used for the design of brassinin oxidase inhibitors (i.e., paldoxins, phytoalexin detoxification inhibitors) because camalexin is a phytoalexin not produced by the Brassica species and L. maculans is unable to metabolize it. The inhibitory effect of camalexin and derivatives decreased as follows: 5-methoxycamalexin > 5-fluorocamalexin = 6-methoxycamalexin > camalexin > 6-fluorocamalexin; 5-methoxycamalexin was determined to be the best inhibitor of brassinin oxidase discovered to date. In addition, the results suggested that camalexin might induce fungal pathways protecting L. maculans against oxidative stress (induction of superoxide dismutase) as well as brassinin toxicity (induction of brassinin oxidase). Overall, these results revealed additional biological effects of camalexin and its natural derivatives and emphasized that different phytoalexins could have positive or negative impacts on plant resistance to different fungal pathogens.

  18. Influence of fungal morphology on the performance of industrial fermentation processes for enzyme production

    DEFF Research Database (Denmark)

    Quintanilla Hernandez, Daniela Alejandra

    at predicting mycelial fragmentation across scales, compared to the EDCF. Furthermore, the morphological development of an industrial strain of T. reesei was monitored in pilot scale fermentations. This study showed that the morphology monitored with laser diffraction also granted the possibility to study......Production of industrial enzymes is usually carried out as submerged aerobic fermentations. Filamentous microorganisms are widely used as hosts in these processes due to multiple advantages. Nevertheless, they also present major drawbacks, due to the unavoidable oxygen transfer limitations...... in this work, along with its correlation to viscosity and other process variables. Considerable research work has been conducted through the years to study fungal morphology and its relation to productivity. However, the work reported in the literature lacks relevant industrial data. In this work, a platform...

  19. 21 CFR 184.1148 - Bacterially-derived carbohydrase enzyme preparation.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Bacterially-derived carbohydrase enzyme preparation... carbohydrase enzyme preparation. (a) Bacterially-derived carbohydrase enzyme preparation is obtained from the... Bacillus subtilis or B. amyloliquefaciens. The preparation is characterized by the presence of the enzymes...

  20. 21 CFR 184.1150 - Bacterially-derived protease enzyme preparation.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Bacterially-derived protease enzyme preparation... enzyme preparation. (a) Bacterially-derived protease enzyme preparation is obtained from the culture... subtilis or B. amyloliquefaciens. The preparation is characterized by the presence of the enzymes...

  1. Biochemical Control of Fungal Biomass and Enzyme Production During Native Hawaiian Litter Degradation

    Science.gov (United States)

    Amatangelo, K. L.; Cordova, T. P.; Vitousek, P. M.

    2007-12-01

    Microbial growth and enzyme production during decomposition is controlled by the availability of carbon substrates, essential elements, and the ratios of these (such as lignin:N). We manipulated carbon:nutrient stoichiometry during decomposition using a natural fertility gradient in Hawaii and litter of varying initial biochemistry. We collected freshly senesced litter of seven biochemically distinct species from three sites offering differing levels of N, P, cations, and 15N , but similar yearly rainfall and temperature patterns. Litter types were decomposed at both the sites they were collected, and at the other site(s) that species was found. Litter was collected at multiple time points, and after one year of decomposition, calculated K constants varied an order of magnitude, from 0.276 to 2.76. Decomposition rates varied significantly with both litter site of origin and deployment, except at the oldest, P-limited site, where litter site of origin was not significantly correlated with decomposition within species. As microbial exocellular enzymes provide the catalyst for the breakdown of organic molecules including phenols, cellulose, and cutin, we assayed polyphenol oxidase, cellobiohydrolase, cutinase, chitinase, and lignin peroxidase to evaluate the breakdown sequence of different litter types. To measure the fungal biomass accumulating during decomposition, we extracted (22E)-Ergosta-5,7,22-trien-3beta- ol (ergosterol) on a subset of samples. The production of particular exocellular enzymes on litter species responded distinctly to origin and decomposition sites: after six months, chitinase and cellobiohydrolase were significantly affected by origin site, whereas polyphenol oxidase activity was controlled by deployment site. We conclude that site characteristics can alter the interaction between litter carbon:nutrient ratios and decomposition rate, mediated through microbial biomass and enzyme production.

  2. Enhancement of fungal pectinolytic enzymes production using gamma radiation under solid state fermentation

    International Nuclear Information System (INIS)

    Ibrahim, Sh.A.

    2013-01-01

    fungal species were screened for their ability to produce pectinases on sugar-beet pulp medium. The highest producer strain was identified as Penicillium citrinum. The optimum conditions for polygalacturonases production were achieved by growing the fungus on sugar beet pulp mineral salts medium and incubation for 7 days at 250 degree C, ph 5.5 and 0.04 g N/g dry SBP by using the conventional method and 1.2 % of nitrogen source by using the factorial design method and surfactant of 0.1% Tween 40. The use of gamma irradiation at a dose of 0.7% kGy yields the highest increase of production of PGase. Polygalacturonases were precipitated from culture supernatant using ammonium sulphate then purified by gel filtration chromatography on sephadex G-100. The optimum ph and temperature of the enzyme activity production were found to be 6.0 and 40°C respectively. The enzyme was found to be stable at ph rang 4 – 8 and showed high stability at temperature rang 20°C -60°C. Mg +2 and Zn +2 stimulated PGase activity.

  3. The preparation and function of the hypertensin-converting enzyme.

    Science.gov (United States)

    SKEGGS, L T; KAHN, J R; SHUMWAY, N P

    1956-03-01

    It has been shown by use of isolated, perfused rat kidneys that hypertensin II is a potent vasoconstrictor substance while hypertensin I is not. Hence it would appear that in intact animals the pressor activity of hypertensin I results from its rapid conversion to hypertensin II. An enzyme which effects this conversion has been procured from horse plasma in a semipurified form by means of ammonium sulfate fractionation and isoelectric precipitation. A method is described for estimating the activity of the enzyme. An example of the use of the preparation in converting purified hypertensin I to hypertensin II has been described.

  4. Enzymes in Commercial Cellulase Preparations Bind Differently to Dioxane Extracted Lignins

    Energy Technology Data Exchange (ETDEWEB)

    Yarbrough, John M.; Mittal, Ashutosh; Katahira, Rui; Mansfield, Elisabeth; Taylor, Larry E.; Decker, Stephen R.; Himmel, Michael E.; Vinzant, Todd

    2017-04-24

    Commercial fungal cellulases used in biomass-to-biofuels processes can be grouped into three general classes: native, augmented, and engineered. To evaluate lignin binding affinities of different enzyme activities in various commercial cellulase formulations in order to determine if enzyme losses due to lignin binding can be modulated by using different enzymes of the same activity We used water:dioxane (1:9) to extract lignin from pretreated corn stover. Commercial cellulases were incubated with lignin and the unbound supernatants were evaluated for individual enzyme loss by SDS=PAGE and these were correlated with activity loss using various pNP-sugar substrates. Colorimetric assays for general glycosyl hydrolase activities showed distinct differences in enzyme binding to lignin for each enzyme activity. Native systems demonstrated low binding of endo- and exo-cellulases, high binding of xylanase, and moderate ..beta..-glucosidase binding. Engineered cellulase mixtures exhibited low binding of exo-cellulases, very strong binding of endocellulases and ..beta..- glucosidase, and mixed binding of xylanase activity. The augmented cellulase had low binding of exocellulase, high binding of endocellulase and xylanase, and moderate binding of ..beta..-glucosidase activities. Bound and unbound activities were correlated with general molecular weight ranges of proteins as measured by loss of proteins bands in bound fractions on SDS-PAGE gels. Lignin-bound high molecular weight bands correlated with binding of ..beta..-glucosidase activity. While ..beta..-glucosidases demonstrated high binding in many cases, they have been shown to remain active. Bound low molecular weight bands correlated with xylanase activity binding. Contrary to other literature, exocellulase activity did not show strong lignin binding. The variation in enzyme activity binding between the three classes of cellulases preparations indicate that it is certainly possible to alter the binding of specific

  5. Several genes encoding enzymes with the same activity are necessary for aerobic fungal degradation of cellulose in nature

    DEFF Research Database (Denmark)

    Busk, Peter Kamp; Lange, Mette; Pilgaard, Bo

    2014-01-01

    feature as it provides a direct route to predict function from primary sequence. Furthermore, we used Peptide Pattern Recognition to compare the cellulose-degrading enzyme activities encoded by 39 fungal genomes. The results indicated that cellobiohydrolases and AA9 lytic polysaccharide monooxygenases....... In the present study we further developed the method Peptide Pattern Recognition to an automatic approach not only to find all genes encoding glycoside hydrolases and lytic polysaccharide monooxygenases in fungal genomes but also to predict the function of the genes. The functional annotation is an important...

  6. Cell Wall Remodeling Enzymes Modulate Fungal Cell Wall Elasticity and Osmotic Stress Resistance.

    Science.gov (United States)

    Ene, Iuliana V; Walker, Louise A; Schiavone, Marion; Lee, Keunsook K; Martin-Yken, Hélène; Dague, Etienne; Gow, Neil A R; Munro, Carol A; Brown, Alistair J P

    2015-07-28

    The fungal cell wall confers cell morphology and protection against environmental insults. For fungal pathogens, the cell wall is a key immunological modulator and an ideal therapeutic target. Yeast cell walls possess an inner matrix of interlinked β-glucan and chitin that is thought to provide tensile strength and rigidity. Yeast cells remodel their walls over time in response to environmental change, a process controlled by evolutionarily conserved stress (Hog1) and cell integrity (Mkc1, Cek1) signaling pathways. These mitogen-activated protein kinase (MAPK) pathways modulate cell wall gene expression, leading to the construction of a new, modified cell wall. We show that the cell wall is not rigid but elastic, displaying rapid structural realignments that impact survival following osmotic shock. Lactate-grown Candida albicans cells are more resistant to hyperosmotic shock than glucose-grown cells. We show that this elevated resistance is not dependent on Hog1 or Mkc1 signaling and that most cell death occurs within 10 min of osmotic shock. Sudden decreases in cell volume drive rapid increases in cell wall thickness. The elevated stress resistance of lactate-grown cells correlates with reduced cell wall elasticity, reflected in slower changes in cell volume following hyperosmotic shock. The cell wall elasticity of lactate-grown cells is increased by a triple mutation that inactivates the Crh family of cell wall cross-linking enzymes, leading to increased sensitivity to hyperosmotic shock. Overexpressing Crh family members in glucose-grown cells reduces cell wall elasticity, providing partial protection against hyperosmotic shock. These changes correlate with structural realignment of the cell wall and with the ability of cells to withstand osmotic shock. The C. albicans cell wall is the first line of defense against external insults, the site of immune recognition by the host, and an attractive target for antifungal therapy. Its tensile strength is conferred by

  7. 21 CFR 184.1420 - Lipase enzyme preparation derived from Rhizopus niveus.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Lipase enzyme preparation derived from Rhizopus... GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1420 Lipase enzyme preparation derived from Rhizopus niveus. (a) Lipase enzyme preparation contains lipase enzyme (CAS Reg. No...

  8. Several genes encoding enzymes with the same activity are necessary for aerobic fungal degradation of cellulose in nature.

    Directory of Open Access Journals (Sweden)

    Peter K Busk

    Full Text Available The cellulose-degrading fungal enzymes are glycoside hydrolases of the GH families and lytic polysaccharide monooxygenases. The entanglement of glycoside hydrolase families and functions makes it difficult to predict the enzymatic activity of glycoside hydrolases based on their sequence. In the present study we further developed the method Peptide Pattern Recognition to an automatic approach not only to find all genes encoding glycoside hydrolases and lytic polysaccharide monooxygenases in fungal genomes but also to predict the function of the genes. The functional annotation is an important feature as it provides a direct route to predict function from primary sequence. Furthermore, we used Peptide Pattern Recognition to compare the cellulose-degrading enzyme activities encoded by 39 fungal genomes. The results indicated that cellobiohydrolases and AA9 lytic polysaccharide monooxygenases are hallmarks of cellulose-degrading fungi except brown rot fungi. Furthermore, a high number of AA9, endocellulase and β-glucosidase genes were identified, not in what are known to be the strongest, specialized lignocellulose degraders but in saprophytic fungi that can use a wide variety of substrates whereas only few of these genes were found in fungi that have a limited number of natural, lignocellulotic substrates. This correlation suggests that enzymes with different properties are necessary for degradation of cellulose in different complex substrates. Interestingly, clustering of the fungi based on their predicted enzymes indicated that Ascomycota and Basidiomycota use the same enzymatic activities to degrade plant cell walls.

  9. The fungal cultivar of leaf-cutter ants produces specific enzymes in response to different plant substrates

    Energy Technology Data Exchange (ETDEWEB)

    Khadempour, Lily [Department of Bacteriology, University of Wisconsin-Madison, Madison WI 53706 USA; Department of Zoology, University of Wisconsin-Madison, Madison WI 53706 USA; Department of Energy Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison WI 53706 USA; Burnum-Johnson, Kristin E. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA 99352 USA; Baker, Erin S. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA 99352 USA; Nicora, Carrie D. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA 99352 USA; Webb-Robertson, Bobbie-Jo M. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA 99352 USA; White, Richard A. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA 99352 USA; Monroe, Matthew E. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA 99352 USA; Huang, Eric L. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA 99352 USA; Smith, Richard D. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA 99352 USA; Currie, Cameron R. [Department of Bacteriology, University of Wisconsin-Madison, Madison WI 53706 USA; Department of Energy Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison WI 53706 USA

    2016-10-26

    Herbivores use symbiotic microbes to help gain access to energy and nutrients from plant material. Leaf-cutter ants are a paradigmatic example, having tremendous impact on their ecosystems as dominant generalist herbivores through cultivation of a fungus, Leucoagaricus gongylophorous. Here we examine how this mutualism could facilitate the flexible substrate incorporation of the ants by providing leaf-cutter ant subcolonies four substrate types: leaves, flowers, oats, and a mixture of all three. Through metaproteomic analysis of the fungus gardens, we were able to identify and quantify 1766 different fungal proteins, including 161 biomass-degrading enzymes. This analysis revealed that fungal protein profiles were significantly different between subcolonies fed different substrates with the highest abundance of cellulolytic enzymes observed in the leaf and flower treatments. When the fungus garden is provided with leaves and flowers, which contain the majority of their energy in recalcitrant material, it increases its production of proteins that break down cellulose: endoglucanases, exoglucanase and β-glucosidase. Further, the complete metaproteomes for the leaves and flowers treatments were very similar, the mixed treatment closely resembled the treatment with oats alone. This suggests that when provided a mixture of substrates, the fungus garden preferentially produces enzymes necessary for breakdown of simpler, more digestible substrates. This flexible, substrate-specific response of the fungal cultivar allows the leaf-cutter ants to derive energy from a wide range of substrates, which may contribute to their ability to be dominant generalist herbivores.

  10. Electron beam technology for production of preparations of immobilized enzymes

    International Nuclear Information System (INIS)

    Gonchar, A.M.; Auslender, V.L.; Polyakov, V.A.

    1995-01-01

    Possibility of electron beam usage for proteases immobilization on 1,4-polyalkylene oxide (1,4-PAO) was studied to obtain biologically active complex for multi-purpose usage. It is shown that immobilization of Bacillus Subtilis protease is done due to free-radical linking of enzyme and carrier with formation of mycelium-like structures. Immobilization improves heat resistance of enzyme up to 60 centigrade without substrate and up to 80 centigrade in presence of substrate, widens range pH activity in comparison with non-immobilized forms. Immobilized proteases does not contain peroxides and long-live radicals. Our results permitted to create technologies for production of medical and veterinary preparations, active components for wool washing agents and leather fabrication technology

  11. 21 CFR 184.1250 - Cellulase enzyme preparation derived from Trichoderma longibrachiatum.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Cellulase enzyme preparation derived from....1250 Cellulase enzyme preparation derived from Trichoderma longibrachiatum. (a) Cellulase enzyme.... reesei). The enzyme, cellulase, catalyzes the endohydrolysis of 1,4-beta-glycosidic linkages in cellulose...

  12. 21 CFR 173.357 - Materials used as fixing agents in the immobilization of enzyme preparations.

    Science.gov (United States)

    2010-04-01

    ... immobilization of enzyme preparations. 173.357 Section 173.357 Food and Drugs FOOD AND DRUG ADMINISTRATION... used as fixing agents in the immobilization of enzyme preparations. Fixing agents may be safely used in the immobilization of enzyme preparations in accordance with the following conditions: (a) The...

  13. 21 CFR 184.1924 - Urease enzyme preparation from Lactobacillus fermentum.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Urease enzyme preparation from Lactobacillus... GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1924 Urease enzyme preparation from Lactobacillus fermentum. (a) This enzyme preparation is derived from the nonpathogenic...

  14. 21 CFR 184.1387 - Lactase enzyme preparation from Candida pseudotropicalis.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Lactase enzyme preparation from Candida... GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1387 Lactase enzyme preparation from Candida pseudotropicalis. (a) This enzyme preparation is derived from the nonpathogenic...

  15. 21 CFR 184.1388 - Lactase enzyme preparation from Kluyveromyces lactis.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Lactase enzyme preparation from Kluyveromyces... GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1388 Lactase enzyme preparation from Kluyveromyces lactis. (a) This enzyme preparation is derived from the nonpathogenic...

  16. 21 CFR 184.1012 - α-Amylase enzyme preparation from Bacillus stearothermophilus.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true α-Amylase enzyme preparation from Bacillus... GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1012 α-Amylase enzyme preparation from Bacillus stearothermophilus. (a) α-Amylase enzyme preparation is obtained from the culture...

  17. Imaging of Bacterial and Fungal Cells Using Fluorescent Carbon Dots Prepared from Carica papaya Juice.

    Science.gov (United States)

    Kasibabu, Betha Saineelima B; D'souza, Stephanie L; Jha, Sanjay; Kailasa, Suresh Kumar

    2015-07-01

    In this paper, we have described a simple hydrothermal method for preparation of fluorescent carbon dots (C-dots) using Carica papaya juice as a precursor. The synthesized C-dots show emission peak at 461 nm with a quantum yield of 7.0 %. The biocompatible nature of C-dots was confirmed by a cytotoxicity assay on E. coli. The C-dots were used as fluorescent probes for imaging of bacterial (Bacillus subtilis) and fungal (Aspergillus aculeatus) cells and emitted green and red colors under different excitation wavelengths, which indicates that the C-dots can be used as a promising material for cell imaging.

  18. Differences in forage-acquisition and fungal enzyme activity contribute to niche segregation in Panamanian leaf-cutting ants.

    Directory of Open Access Journals (Sweden)

    Pepijn W Kooij

    Full Text Available The genera Atta and Acromyrmex are often grouped as leaf-cutting ants for pest management assessments and ecological surveys, although their mature colony sizes and foraging niches may differ substantially. Few studies have addressed such interspecific differences at the same site, which prompted us to conduct a comparative study across six sympatric leaf-cutting ant species in Central Panama. We show that foraging rates during the transition between dry and wet season differ about 60 fold between genera, but are relatively constant across species within genera. These differences appear to match overall differences in colony size, especially when Atta workers that return to their nests without leaves are assumed to carry liquid food. We confirm that Panamanian Atta specialize primarily on tree-leaves whereas Acromyrmex focus on collecting flowers and herbal leaves and that species within genera are similar in these overall foraging strategies. Species within genera tended to be spaced out over the three habitat categories that we distinguished (forest, forest edge, open grassland, but each of these habitats normally had only a single predominant Atta and Acromyrmex species. We measured activities of twelve fungus garden decomposition enzymes, belonging to the amylases, cellulases, hemicellulases, pectinases and proteinases, and show that average enzyme activity per unit of fungal mass in Atta gardens is lower than in Acromyrmex gardens. Expression profiles of fungal enzymes in Atta also appeared to be more specialized than in Acromyrmex, possibly reflecting variation in forage material. Our results suggest that species- and genus-level identities of leaf-cutting ants and habitat-specific foraging profiles may give predictable differences in the expression of fungal genes coding for decomposition enzymes.

  19. Differences in Forage-Acquisition and Fungal Enzyme Activity Contribute to Niche Segregation in Panamanian Leaf-Cutting Ants

    Science.gov (United States)

    Kooij, Pepijn W.; Liberti, Joanito; Giampoudakis, Konstantinos; Schiøtt, Morten; Boomsma, Jacobus J.

    2014-01-01

    The genera Atta and Acromyrmex are often grouped as leaf-cutting ants for pest management assessments and ecological surveys, although their mature colony sizes and foraging niches may differ substantially. Few studies have addressed such interspecific differences at the same site, which prompted us to conduct a comparative study across six sympatric leaf-cutting ant species in Central Panama. We show that foraging rates during the transition between dry and wet season differ about 60 fold between genera, but are relatively constant across species within genera. These differences appear to match overall differences in colony size, especially when Atta workers that return to their nests without leaves are assumed to carry liquid food. We confirm that Panamanian Atta specialize primarily on tree-leaves whereas Acromyrmex focus on collecting flowers and herbal leaves and that species within genera are similar in these overall foraging strategies. Species within genera tended to be spaced out over the three habitat categories that we distinguished (forest, forest edge, open grassland), but each of these habitats normally had only a single predominant Atta and Acromyrmex species. We measured activities of twelve fungus garden decomposition enzymes, belonging to the amylases, cellulases, hemicellulases, pectinases and proteinases, and show that average enzyme activity per unit of fungal mass in Atta gardens is lower than in Acromyrmex gardens. Expression profiles of fungal enzymes in Atta also appeared to be more specialized than in Acromyrmex, possibly reflecting variation in forage material. Our results suggest that species- and genus-level identities of leaf-cutting ants and habitat-specific foraging profiles may give predictable differences in the expression of fungal genes coding for decomposition enzymes. PMID:24718261

  20. Enhanced oxidation of benzo[a]pyrene by crude enzyme extracts produced during interspecific fungal interaction of Trametes versicolor and Phanerochaete chrysosporium.

    Science.gov (United States)

    Qian, Linbo; Chen, Baoliang

    2012-01-01

    The effects of interspecific fungal interactions between Trametes versicolor and Phanerochaete chrysosporium on laccase activity and enzymatic oxidation of polycyclic aromatic hydrocarbons (PAHs) were investigated. A deadlock between the two mycelia rather than replacement of one fungus by another was observed on an agar medium. The laccase activity in crude enzyme extracts from interaction zones reached a maximum after a 5-day incubation, which was significantly higher than that from regions of T. versicolor or P. chrysosporium alone. The enhanced induction of laccase activity lasted longer in half nutrition than in normal nutrition. A higher potential to oxidize benzo[a]pyrene by a crude enzyme preparation extracted from the interaction zones was demonstrated. After a 48 hr incubation period, the oxidation of benzo[a]pyrene by crude enzyme extracts from interaction zones reached 26.2%, while only 9.5% of benzo[a]pyrene was oxidized by crude extracts from T. versicolor. The oxidation was promoted by the co-oxidant 2,2'-azinobis-3-ethylbenzthiazoline-6-sulphonate diammonium salt (ABTS). These findings indicate that the application of co-culturing of white-rot fungi in bioremediation is a potential ameliorating technique for the restoration of PAH-contaminated soil.

  1. Using an Inducible Promoter of a Gene Encoding Penicillium verruculosum Glucoamylase for Production of Enzyme Preparations with Enhanced Cellulase Performance.

    Directory of Open Access Journals (Sweden)

    Alexander G Bulakhov

    Full Text Available Penicillium verruculosum is an efficient producer of highly active cellulase multienzyme system. One of the approaches for enhancing cellulase performance in hydrolysis of cellulosic substrates is to enrich the reaction system with β -glucosidase and/or accessory enzymes, such as lytic polysaccharide monooxygenases (LPMO displaying a synergism with cellulases.Genes bglI, encoding β-glucosidase from Aspergillus niger (AnBGL, and eglIV, encoding LPMO (formerly endoglucanase IV from Trichoderma reesei (TrLPMO, were cloned and expressed by P. verruculosum B1-537 strain under the control of the inducible gla1 gene promoter. Content of the heterologous AnBGL in the secreted multienzyme cocktails (hBGL1, hBGL2 and hBGL3 varied from 4 to 10% of the total protein, while the content of TrLPMO in the hLPMO sample was ~3%. The glucose yields in 48-h hydrolysis of Avicel and milled aspen wood by the hBGL1, hBGL2 and hBGL3 preparations increased by up to 99 and 80%, respectively, relative to control enzyme preparations without the heterologous AnBGL (at protein loading 5 mg/g substrate for all enzyme samples. The heterologous TrLPMO in the hLPMO preparation boosted the conversion of the lignocellulosic substrate by 10-43%; however, in hydrolysis of Avicel the hLPMO sample was less effective than the control preparations. The highest product yield in hydrolysis of aspen wood was obtained when the hBGL2 and hLPMO preparations were used at the ratio 1:1.The enzyme preparations produced by recombinant P. verruculosum strains, expressing the heterologous AnBGL or TrLPMO under the control of the gla1 gene promoter in a starch-containing medium, proved to be more effective in hydrolysis of a lignocellulosic substrate than control enzyme preparations without the heterologous enzymes. The enzyme composition containing both AnBGL and TrLPMO demonstrated the highest performance in lignocellulose hydrolysis, providing a background for developing a fungal strain capable

  2. The fungal cultivar of leaf-cutter ants produces specific enzymes in response to different plant substrates.

    Science.gov (United States)

    Khadempour, Lily; Burnum-Johnson, Kristin E; Baker, Erin S; Nicora, Carrie D; Webb-Robertson, Bobbie-Jo M; White, Richard A; Monroe, Matthew E; Huang, Eric L; Smith, Richard D; Currie, Cameron R

    2016-11-01

    Herbivores use symbiotic microbes to help derive energy and nutrients from plant material. Leaf-cutter ants are a paradigmatic example, cultivating their mutualistic fungus Leucoagaricus gongylophorus on plant biomass that workers forage from a diverse collection of plant species. Here, we investigate the metabolic flexibility of the ants' fungal cultivar for utilizing different plant biomass. Using feeding experiments and a novel approach in metaproteomics, we examine the enzymatic response of L. gongylophorus to leaves, flowers, oats or a mixture of all three. Across all treatments, our analysis identified and quantified 1766 different fungal proteins, including 161 putative biomass-degrading enzymes. We found significant differences in the protein profiles in the fungus gardens of subcolonies fed different plant substrates. When provided with leaves or flowers, which contain the majority of their energy as recalcitrant plant polymers, the fungus gardens produced more proteins predicted to break down cellulose: endoglucanase, exoglucanase and β-glucosidase. Further, the complete metaproteomes for the leaves and flowers treatments were very similar, while the mixed substrate treatment closely resembled the treatment with oats alone. This indicates that when provided a mixture of plant substrates, fungus gardens preferentially break down the simpler, more digestible substrates. This flexible, substrate-specific enzymatic response of the fungal cultivar allows leaf-cutter ants to derive energy from a wide range of substrates, which likely contributes to their ability to be dominant generalist herbivores. © 2016 John Wiley & Sons Ltd.

  3. Efficient screening of fungal cellobiohydrolase class I enzymes for thermostabilizing sequence blocks by SCHEMA structure-guided recombination.

    Science.gov (United States)

    Heinzelman, Pete; Komor, Russell; Kanaan, Arvind; Romero, Philip; Yu, Xinlin; Mohler, Shannon; Snow, Christopher; Arnold, Frances

    2010-11-01

    We describe an efficient SCHEMA recombination-based approach for screening homologous enzymes to identify stabilizing amino acid sequence blocks. This approach has been used to generate active, thermostable cellobiohydrolase class I (CBH I) enzymes from the 390 625 possible chimeras that can be made by swapping eight blocks from five fungal homologs. Constructing and characterizing the parent enzymes and just 32 'monomeras' containing a single block from a homologous enzyme allowed stability contributions to be assigned to 36 of the 40 blocks from which the CBH I chimeras can be assembled. Sixteen of 16 predicted thermostable chimeras, with an average of 37 mutations relative to the closest parent, are more thermostable than the most stable parent CBH I, from the thermophilic fungus Talaromyces emersonii. Whereas none of the parent CBH Is were active >65°C, stable CBH I chimeras hydrolyzed solid cellulose at 70°C. In addition to providing a collection of diverse, thermostable CBH Is that can complement previously described stable CBH II chimeras (Heinzelman et al., Proc. Natl Acad. Sci. USA 2009;106:5610-5615) in formulating application-specific cellulase mixtures, the results show the utility of SCHEMA recombination for screening large swaths of natural enzyme sequence space for desirable amino acid blocks.

  4. Flavourzyme, an Enzyme Preparation with Industrial Relevance: Automated Nine-Step Purification and Partial Characterization of Eight Enzymes.

    Science.gov (United States)

    Merz, Michael; Eisele, Thomas; Berends, Pieter; Appel, Daniel; Rabe, Swen; Blank, Imre; Stressler, Timo; Fischer, Lutz

    2015-06-17

    Flavourzyme is sold as a peptidase preparation from Aspergillus oryzae. The enzyme preparation is widely and diversely used for protein hydrolysis in industrial and research applications. However, detailed information about the composition of this mixture is still missing due to the complexity. The present study identified eight key enzymes by mass spectrometry and partially by activity staining on native polyacrylamide gels or gel zymography. The eight enzymes identified were two aminopeptidases, two dipeptidyl peptidases, three endopeptidases, and one α-amylase from the A. oryzae strain ATCC 42149/RIB 40 (yellow koji mold). Various specific marker substrates for these Flavourzyme enzymes were ascertained. An automated, time-saving nine-step protocol for the purification of all eight enzymes within 7 h was designed. Finally, the purified Flavourzyme enzymes were biochemically characterized with regard to pH and temperature profiles and molecular sizes.

  5. Milk clotting and proteolytic activity of enzyme preparation from ...

    African Journals Online (AJOL)

    Some microorganisms have the ability to produce enzymes that could clot milk and used as a substitute for calf rennet. Strains of lactic acid bacteria (LAB) could produce proteolytic enzymes that may have the potential to be used as a source of milk clotting enzyme (MCE). In the present study, LAB isolated from shrimp paste ...

  6. The phytoalexins brassilexin and camalexin inhibit cyclobrassinin hydrolase, a unique enzyme from the fungal pathogen Alternaria brassicicola.

    Science.gov (United States)

    Pedras, M Soledade C; Minic, Zoran

    2014-01-01

    Alternaria brassicicola is a fungal pathogen of many agriculturally important cruciferous crops. Cyclobrassinin hydrolase (CH) is an enzyme produced by A. brassicicola that catalyzes the transformation of the cruciferous phytoalexin cyclobrassinin into S-methyl[(2-sulfanyl-1H-indolyl-3)methyl]carbamothioate. The purification and characterization of CH was performed using a four-step chromatography method. SDS-PAGE and gel exclusion chromatography indicated that CH is a tetrameric protein with molecular mass of 330 kDa. Sequence analysis and chemical modification of CH with selective reagents suggested that the enzyme mediates hydrolysis of cyclobrassinin using a catalytic amino acid triad. Enzyme kinetic studies using cyclobrassinin and 1-methylcyclobrassinin as substrates revealed that CH displayed positive substrate cooperativity. Investigation of the effect of nine phytoalexins and two derivatives on the activity of CH indicated that six compounds displayed inhibitory activity: brassilexin, 1-methylbrassilexin, dioxibrassinin, camalexin, brassicanal A and sinalexin. The enzyme kinetics of CH strongly suggested that brassilexin and 1-methylbrassilexin are noncompetitive inhibitors of CH activity, and that camalexin is a competitive inhibitor while dioxibrassinin inhibits CH through a mixed mechanism. The phytoalexin brassilexin is the most effective inhibitor of CH (K(i)=32 ± 9 μM). These results suggest that crops able to accumulate higher concentration of brassilexin would display higher resistance levels to the fungus. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Variation in fungal enzyme spectra may affect mutualistic division of labour between ants and fungus gardens

    DEFF Research Database (Denmark)

    de Fine Licht, Henrik Hjarvard; Boomsma, Jacobus Jan

    partners vary in metabolic performance, division of labour may not always be optimized and co-evolutionary trajectories become less predictable. The higher fungus-growing (attine) ants consist of the leafcutter ants (Acromyrmex and Atta), which rear a single fungal species throughout their Latin American...... to be analogous to a diverse array of subsistence farming practices at the mercy of local conditions, whereas the latter resemble large-scale, low-diversity “industrial” farming....

  8. Fungal Enzymes and Yeasts for Conversion of Plant Biomass to Bioenergy and High-Value Products

    DEFF Research Database (Denmark)

    Lange, Lene

    2017-01-01

    in the conversion of plant biomass to value-added products. These products provide a basis for substituting fossil-derived fuels, chemicals, and materials, as well as unlocking the biomass potential of the agricultural harvest to yield more food and feed. This article focuses on the mycological basis for the fungal...... contributed to mycology and environmental research? Future perspectives and approaches are listed, highlighting the importance of fungi in development of the bioeconomy....

  9. Application of residual polysaccharide-degrading enzymes in dried shiitake mushrooms as an enzyme preparation in food processing.

    Science.gov (United States)

    Tatsumi, E; Konishi, Y; Tsujiyama, S

    2016-11-01

    To examine the activities of residual enzymes in dried shiitake mushrooms, which are a traditional foodstuff in Japanese cuisine, for possible applications in food processing. Polysaccharide-degrading enzymes remained intact in dried shiitake mushrooms and the activities of amylase, β-glucosidase and pectinase were high. A potato digestion was tested using dried shiitake powder. The enzymes reacted with potato tuber specimens to solubilize sugars even under a heterogeneous solid-state condition and that their reaction modes were different at 38 and 50 °C. Dried shiitake mushrooms have a potential use in food processing as an enzyme preparation.

  10. Effort versus Reward: Preparing Samples for Fungal Community Characterization in High-Throughput Sequencing Surveys of Soils.

    Directory of Open Access Journals (Sweden)

    Zewei Song

    Full Text Available Next generation fungal amplicon sequencing is being used with increasing frequency to study fungal diversity in various ecosystems; however, the influence of sample preparation on the characterization of fungal community is poorly understood. We investigated the effects of four procedural modifications to library preparation for high-throughput sequencing (HTS. The following treatments were considered: 1 the amount of soil used in DNA extraction, 2 the inclusion of additional steps (freeze/thaw cycles, sonication, or hot water bath incubation in the extraction procedure, 3 the amount of DNA template used in PCR, and 4 the effect of sample pooling, either physically or computationally. Soils from two different ecosystems in Minnesota, USA, one prairie and one forest site, were used to assess the generality of our results. The first three treatments did not significantly influence observed fungal OTU richness or community structure at either site. Physical pooling captured more OTU richness compared to individual samples, but total OTU richness at each site was highest when individual samples were computationally combined. We conclude that standard extraction kit protocols are well optimized for fungal HTS surveys, but because sample pooling can significantly influence OTU richness estimates, it is important to carefully consider the study aims when planning sampling procedures.

  11. 21 CFR 184.1985 - Aminopeptidase enzyme preparation derived from lactococcus lactis.

    Science.gov (United States)

    2010-04-01

    ... lactococcus lactis. 184.1985 Section 184.1985 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF... Aminopeptidase enzyme preparation derived from lactococcus lactis. (a) Aminopeptidase enzyme preparation is derived from the nonpathogenic and nontoxicogenic bacterium Lactococcus lactis (previously named...

  12. Analysis of the Sequences, Structures, and Functions of Product-Releasing Enzyme Domains in Fungal Polyketide Synthases

    Directory of Open Access Journals (Sweden)

    Lu Liu

    2017-09-01

    Full Text Available Product-releasing enzyme (PRE domains in fungal non-reducing polyketide synthases (NR-PKSs play a crucial role in catalysis and editing during polyketide biosynthesis, especially accelerating final biosynthetic reactions accompanied with product offloading. However, up to date, the systematic knowledge about PRE domains is deficient. In the present study, the relationships between sequences, structures, and functions of PRE domains were analyzed with 574 NR-PKSs of eight groups (I–VIII. It was found that the PRE domains in NR-PKSs could be mainly classified into three types, thioesterase (TE, reductase (R, and metallo-β-lactamase-type TE (MβL-TE. The widely distributed TE or TE-like domains were involved in NR-PKSs of groups I–IV, VI, and VIII. The R domains appeared in NR-PKSs of groups IV and VII, while the physically discrete MβL-TE domains were employed by most NR-PKSs of group V. The changes of catalytic sites and structural characteristics resulted in PRE functional differentiations. The phylogeny revealed that the evolution of TE domains was accompanied by complex functional divergence. The diverse sequence lengths of TE lid-loops affected substrate specificity with different chain lengths. The volume diversification of TE catalytic pockets contributed to catalytic mechanisms with functional differentiations. The above findings may help to understand the crucial catalysis of fungal aromatic polyketide biosyntheses and govern recombination of NR-PKSs to obtain unnatural target products.

  13. Comprehensive analysis of fungal diversity and enzyme activity in nuruk, a Korean fermenting starter, for acquiring useful fungi.

    Science.gov (United States)

    Carroll, Emily; Trinh, Tran Ngoc; Son, Hokyoung; Lee, Yin-Won; Seo, Jeong-Ah

    2017-05-01

    Nuruk is a fermenting starter that is involved in the production of alcoholic beverages, and has been used in South Korea for a very long time. To analyze the fungal diversity, we collected a total of 59 nuruk samples from several companies and persons in 2013 to 2014, and obtained 364 isolates. All of the single isolated fungi were identified, both morphologically and molecularly, based on the sequences of ribosomal RNA gene [18S, ITS1-5.8S-ITS2, and 26S (D1/D2 region)]. In 46 nuruk samples out of 59 (78%), Saccharomycopsis fibuligera, a dimorphic yeast, was most frequently isolated. Among the filamentous fungi, Aspergillus and Lichtheimia were found in more than 50% of the samples with lower colony forming unit (CFU/g of sample) than those of yeasts. The yeasts S. fibuligera and Wickerhamomyces anomalus were counted with maximum 1.3-1.8 × 10 8 CFU/g. Among Mucorales fungi, Lichtheimia and Mucor were isolated in much higher numbers than Rhizopus and Rhizomucor. Overall, the home-made nuruks tend to contain more diverse filamentous fungi than the commercial nuruks. To acquire industrially useful filamentous fungi and yeasts, we analyzed the enzyme activities of α-amylase, glucoamylase and acid protease associated with brewing properties for 131 strains. Aspergillus oryzae and S. fibuligera had high α- and glucoamylase activities and most isolates of Lichtheimia ramosa had high acid protease activity. For further applications, 27 fungal strains were chosen based on isolation frequencies from nuruk, and the ability to produce useful enzyme.

  14. Effect of neohesperidin dihydrochalcone on the activity and stability of alpha-amylase: a comparative study on bacterial, fungal, and mammalian enzymes.

    Science.gov (United States)

    Kashani-Amin, Elaheh; Ebrahim-Habibi, Azadeh; Larijani, Bagher; Moosavi-Movahedi, Ali Akbar

    2015-10-01

    Neohesperidin dihydrochalcone (NHDC) was recently introduced as an activator of mammalian alpha-amylase. In the current study, the effect of NHDC has been investigated on bacterial and fungal alpha-amylases. Enzyme assays and kinetic analysis demonstrated the capability of NHDC to significantly activate both tested alpha-amylases. The ligand activation pattern was found to be more similar between the fungal and mammalian enzyme in comparison with the bacterial one. Further, thermostability experiments indicated a stability increase in the presence of NHDC for the bacterial enzyme. In silico (docking) test locates a putative binding site for NHDC on alpha-amylase surface in domain B. This domain shows differences in various alpha-amylase types, and the different behavior of the ligand toward the studied enzymes may be attributed to this fact. Copyright © 2015 John Wiley & Sons, Ltd.

  15. Enzymes from fungal and plant origin required for chemical diversification of insecticidal loline alkaloids in grass-Epichloë symbiota.

    Directory of Open Access Journals (Sweden)

    Juan Pan

    Full Text Available The lolines are a class of bioprotective alkaloids that are produced by Epichloë species, fungal endophytes of grasses. These alkaloids are saturated 1-aminopyrrolizidines with a C2 to C7 ether bridge, and are structurally differentiated by the various modifications of the 1-amino group: -NH2 (norloline, -NHCH3 (loline, -N(CH32 (N-methylloline, -N(CH3Ac (N-acetylloline, -NHAc (N-acetylnorloline, and -N(CH3CHO (N-formylloline. Other than the LolP cytochrome P450, which is required for conversion of N-methylloline to N-formylloline, the enzymatic steps for loline diversification have not yet been established. Through isotopic labeling, we determined that N-acetylnorloline is the first fully cyclized loline alkaloid, implying that deacetylation, methylation, and acetylation steps are all involved in loline alkaloid diversification. Two genes of the loline alkaloid biosynthesis (LOL gene cluster, lolN and lolM, were predicted to encode an N-acetamidase (deacetylase and a methyltransferase, respectively. A knockout strain lacking both lolN and lolM stopped the biosynthesis at N-acetylnorloline, and complementation with the two wild-type genes restored production of N-formylloline and N-acetylloline. These results indicated that lolN and lolM are required in the steps from N-acetylnorloline to other lolines. The function of LolM as an N-methyltransferase was confirmed by its heterologous expression in yeast resulting in conversion of norloline to loline, and of loline to N-methylloline. One of the more abundant lolines, N-acetylloline, was observed in some but not all plants with symbiotic Epichloë siegelii, and when provided with exogenous loline, asymbiotic meadow fescue (Lolium pratense plants produced N-acetylloline, suggesting that a plant acetyltransferase catalyzes N-acetylloline formation. We conclude that although most loline alkaloid biosynthesis reactions are catalyzed by fungal enzymes, both fungal and plant enzymes are responsible for

  16. Enzymes from Fungal and Plant Origin Required for Chemical Diversification of Insecticidal Loline Alkaloids in Grass-Epichloë Symbiota

    Science.gov (United States)

    Pan, Juan; Bhardwaj, Minakshi; Nagabhyru, Padmaja; Grossman, Robert B.; Schardl, Christopher L.

    2014-01-01

    The lolines are a class of bioprotective alkaloids that are produced by Epichloë species, fungal endophytes of grasses. These alkaloids are saturated 1-aminopyrrolizidines with a C2 to C7 ether bridge, and are structurally differentiated by the various modifications of the 1-amino group: -NH2 (norloline), -NHCH3 (loline), -N(CH3)2 (N-methylloline), -N(CH3)Ac (N-acetylloline), -NHAc (N-acetylnorloline), and -N(CH3)CHO (N-formylloline). Other than the LolP cytochrome P450, which is required for conversion of N-methylloline to N-formylloline, the enzymatic steps for loline diversification have not yet been established. Through isotopic labeling, we determined that N-acetylnorloline is the first fully cyclized loline alkaloid, implying that deacetylation, methylation, and acetylation steps are all involved in loline alkaloid diversification. Two genes of the loline alkaloid biosynthesis (LOL) gene cluster, lolN and lolM, were predicted to encode an N-acetamidase (deacetylase) and a methyltransferase, respectively. A knockout strain lacking both lolN and lolM stopped the biosynthesis at N-acetylnorloline, and complementation with the two wild-type genes restored production of N-formylloline and N-acetylloline. These results indicated that lolN and lolM are required in the steps from N-acetylnorloline to other lolines. The function of LolM as an N-methyltransferase was confirmed by its heterologous expression in yeast resulting in conversion of norloline to loline, and of loline to N-methylloline. One of the more abundant lolines, N-acetylloline, was observed in some but not all plants with symbiotic Epichloë siegelii, and when provided with exogenous loline, asymbiotic meadow fescue (Lolium pratense) plants produced N-acetylloline, suggesting that a plant acetyltransferase catalyzes N-acetylloline formation. We conclude that although most loline alkaloid biosynthesis reactions are catalyzed by fungal enzymes, both fungal and plant enzymes are responsible for the

  17. Enzymes from fungal and plant origin required for chemical diversification of insecticidal loline alkaloids in grass-Epichloë symbiota.

    Science.gov (United States)

    Pan, Juan; Bhardwaj, Minakshi; Nagabhyru, Padmaja; Grossman, Robert B; Schardl, Christopher L

    2014-01-01

    The lolines are a class of bioprotective alkaloids that are produced by Epichloë species, fungal endophytes of grasses. These alkaloids are saturated 1-aminopyrrolizidines with a C2 to C7 ether bridge, and are structurally differentiated by the various modifications of the 1-amino group: -NH2 (norloline), -NHCH3 (loline), -N(CH3)2 (N-methylloline), -N(CH3)Ac (N-acetylloline), -NHAc (N-acetylnorloline), and -N(CH3)CHO (N-formylloline). Other than the LolP cytochrome P450, which is required for conversion of N-methylloline to N-formylloline, the enzymatic steps for loline diversification have not yet been established. Through isotopic labeling, we determined that N-acetylnorloline is the first fully cyclized loline alkaloid, implying that deacetylation, methylation, and acetylation steps are all involved in loline alkaloid diversification. Two genes of the loline alkaloid biosynthesis (LOL) gene cluster, lolN and lolM, were predicted to encode an N-acetamidase (deacetylase) and a methyltransferase, respectively. A knockout strain lacking both lolN and lolM stopped the biosynthesis at N-acetylnorloline, and complementation with the two wild-type genes restored production of N-formylloline and N-acetylloline. These results indicated that lolN and lolM are required in the steps from N-acetylnorloline to other lolines. The function of LolM as an N-methyltransferase was confirmed by its heterologous expression in yeast resulting in conversion of norloline to loline, and of loline to N-methylloline. One of the more abundant lolines, N-acetylloline, was observed in some but not all plants with symbiotic Epichloë siegelii, and when provided with exogenous loline, asymbiotic meadow fescue (Lolium pratense) plants produced N-acetylloline, suggesting that a plant acetyltransferase catalyzes N-acetylloline formation. We conclude that although most loline alkaloid biosynthesis reactions are catalyzed by fungal enzymes, both fungal and plant enzymes are responsible for the

  18. Synergistic action of enzyme preparations towards recalcitrant corn silage polysaccharides

    NARCIS (Netherlands)

    Neumüller, K.G.; Streekstra, H.; Schols, H.A.; Gruppen, H.

    2014-01-01

    Corn silage, its water unextractable solids (WUS) and enzyme recalcitrant solids (ErCS) and an industrial corn silage-based anaerobic fermentation residue (AFR) represent corn substrates with different levels of recalcitrance. Compositional analysis reveals different levels of arabinoxylan

  19. Small-scale distribution of extracellular enzymes, fungal, and bacterial biomass in Quercus petraea forest topsoil

    Czech Academy of Sciences Publication Activity Database

    Baldrian, Petr; Merhautová, Věra; Cajthaml, Tomáš; Petránková, Mirka; Šnajdr, Jaroslav

    2010-01-01

    Roč. 46, č. 7 (2010), s. 717-726 ISSN 0178-2762 R&D Projects: GA MŠk LC06066; GA MŠk(CZ) ME10152; GA MZe QH72216 Institutional research plan: CEZ:AV0Z50200510 Keywords : Decomposition * Extracellular enzymes * Forest soil Subject RIV: EE - Microbiology, Virology Impact factor: 2.156, year: 2010

  20. Diversity and Enzyme Activity of Ectomycorrhizal Fungal Communities Following Nitrogen Fertilization in an Urban-Adjacent Pine Plantation

    Directory of Open Access Journals (Sweden)

    Chen Ning

    2018-02-01

    Full Text Available Rapid economic development and accelerated urbanization in China has resulted in widespread atmospheric nitrogen (N deposition. One consequence of N deposition is the alteration of mycorrhizal symbioses that are critical for plant resource acquisition (nitrogen, N, phosphorus, P, water. In this study, we characterized the diversity, composition, and functioning of ectomycorrhizal (ECM fungal communities in an urban-adjacent Pinus elliottii plantation under ambient N deposition (~24 kg N ha−1 year−1, and following N fertilization (low N, 50 kg N ha−1 year−1; high N, 300 kg N ha−1 year−1. ECM functioning was expressed as the potential activities of extracellular enzymes required for organic N (protease, P (phosphomonoesterase, and recalcitrant polymers (phenol oxidase. Despite high ambient N deposition, ECM community composition shifted under experimental N fertilization, and those changes were linked to disparate levels of soil minerals (P, K and organic matter (but not N, a decline in acid phosphatase (AP, and an increase in phenol oxidase (PO potential activities. Based on enzyme stoichiometry, medium-smooth exploration type ECM species invested more in C acquisition (PO relative to P (AP following high N fertilization than other exploration types. ECM species with hydrophilic mantles also showed higher enzymatic PO:AP ratios than taxa with hydrophobic mantles. Our findings add to the accumulating evidence that shifts in ECM community composition and taxa specialized in organic C, N, and P degradation could modulate the soil nutrient cycling in forests exposed to chronic elevated N input.

  1. Isotope Effects Associated with N2O Production by Fungal and Bacterial Nitric Oxide Reductases: Implications for Enzyme Mechanisms

    Science.gov (United States)

    Hegg, E. L.; Yang, H.; Gandhi, H.; McQuarters, A.; Lehnert, N.; Ostrom, N. E.

    2014-12-01

    Nitrous oxide (N2O) is both a powerful greenhouse gas and a key participant in ozone destruction. Microbial activity accounts for over 70% of the N2O produced annually, and the atmospheric concentration of N2O continues to rise. Because the fungal and bacterial denitrification pathways are major contributors to microbial N2O production, understanding the mechanism by which NO is reduced to N2O will contribute to both N2O source tracing and quantification. Our strategy utilizes stable isotopes to probe the enzymatic mechanism of microbial N2O production. Although the use of stable isotopes to study enzyme mechanisms is not new, our approach is distinct in that we employ both measurements of isotopic preferences of purified enzyme and DFT calculations, thereby providing a synergistic combination of experimental and computational approaches. We analyzed δ18O, δ15Nα (central N atom in N2O), and δ15Nβ (terminal N atom) of N2O produced by purified fungal cytochrome P450 nitric oxide reductase (P450nor) from Histoplasma capsulatum as well as bacterial cytochrome c dependent nitric oxide reductase (cNOR) from Paracoccus denitrificans. P450nor exhibits an inverse kinetic isotope effect for Nβ (KIE = 0.9651) but a normal isotope effect for both Nα (KIE = 1.0127) and the oxygen atom (KIE = 1.0264). These results suggest a mechanism where NO binds to the ferric heme in the P450nor active site and becomes Nβ. Analysis of the NO-binding step indicated a greater difference in zero point energy in the transition state than the ground state, resulting in the inverse KIE observed for Nβ. Following protonation and rearrangement, it is speculated that this complex forms a FeIV-NHOH- species as a key intermediate. Our data are consistent with the second NO (which becomes Nα and O in the N2O product) attacking the FeIV-NHOH- species to generate a FeIII-N2O2H2 complex that enzymatically (as opposed to abiotically) breaks down to release N2O. Conversely, our preliminary data

  2. Enzyme Preparations of the Pancreas without Enteric Coating: Whether they are Used Today?

    Directory of Open Access Journals (Sweden)

    A.V. Savustyanenko

    2014-09-01

    Full Text Available Introduction. For the treatment of exocrine pancreatic insufficiency (EPI it is recommended enzyme replacement therapy with high-dose enteric-coated preparations. In this regard, the question arises about the rationale of clinical application of the older low-dose non enteric-coated enzyme preparations (as a reference drug served Mezim® forte. Results. A number of anatomical and physiological characteristics and active use of suppressants of gastric secretion — proton pump inhibitors, antacids (sodium bicarbonate — allows significantly to increase the «survival» of pancreatic enzymes that do not contain enteric coating in gastric juice and their delivery to the duodenum. This formulation is used for the relief of pain in case of chronic pancreatitis (enteric-coated preparations are uneffective. It was proposed to use a combination of two dosage forms — enteric-coated and non enteric-coated — for more effective therapy EPI. In functional diseases of the gastrointestinal tract (irritable bowel syndrome with functional disorders of the pancreas, functional dyspepsia non enteric-coated enzyme preparations speeds recovery of patients. In inflammatory bowel disease (ulcerative colitis, Crohn’s disease and chronic obstructive pulmonary disease, this formulation increases the duration of remission, and in case of inflammatory bowel disease — also the quality of life during remission. Non enteric-coated enzyme preparations are used in a number of other diseases associated with maldigestion/malabsorption. Additionally, they are used for the gradual cessation of therapy by high-dose enzyme preparations. They also find application for the correction of EPI during snacks and some gaps in the diet. Financial sales figures indicate a high demand for low-dose non enteric-coated enzyme preparations (Mezim® forte. Conclusions. Low-dose non enteric-coated enzyme preparations are still widely used in current clinical practice.

  3. Molecular engineering of fungal GH5 and GH26 beta-(1,4-mannanases toward improvement of enzyme activity.

    Directory of Open Access Journals (Sweden)

    Marie Couturier

    Full Text Available Microbial mannanases are biotechnologically important enzymes since they target the hydrolysis of hemicellulosic polysaccharides of softwood biomass into simple molecules like manno-oligosaccharides and mannose. In this study, we have implemented a strategy of molecular engineering in the yeast Yarrowia lipolytica to improve the specific activity of two fungal endo-mannanases, PaMan5A and PaMan26A, which belong to the glycoside hydrolase (GH families GH5 and GH26, respectively. Following random mutagenesis and two steps of high-throughput enzymatic screening, we identified several PaMan5A and PaMan26A mutants that displayed improved kinetic constants for the hydrolysis of galactomannan. Examination of the three-dimensional structures of PaMan5A and PaMan26A revealed which of the mutated residues are potentially important for enzyme function. Among them, the PaMan5A-G311S single mutant, which displayed an impressive 8.2-fold increase in kcat /KM due to a significant decrease of KM, is located within the core of the enzyme. The PaMan5A-K139R/Y223H double mutant revealed modification of hydrolysis products probably in relation to an amino-acid substitution located nearby one of the positive subsites. The PaMan26A-P140L/D416G double mutant yielded a 30% increase in kcat /KM compared to the parental enzyme. It displayed a mutation in the linker region (P140L that may confer more flexibility to the linker and another mutation (D416G located at the entrance of the catalytic cleft that may promote the entrance of the substrate into the active site. Taken together, these results show that the directed evolution strategy implemented in this study was very pertinent since a straightforward round of random mutagenesis yielded significantly improved variants, in terms of catalytic efiiciency (kcat/KM.

  4. Nuclear magnetic resonance characterization of apple juice containing enzyme preparations

    International Nuclear Information System (INIS)

    Prestes, Rosilene A.; Almeida, Denise Milleo; Barison, Andersson; Pinheiro, Luis Antonio; Wosiacki, Gilvan

    2012-01-01

    In this work, 1 H nuclear magnetic resonance ( 1 H NMR) was employed to evaluate changes in apple juice in response to the addition of Panzym Yieldmash and Ultrazym AFP-L enzymatic complexes and compare it with premium apple juice. The juice was processed at different temperatures and concentrations of enzymatic complexes. The differences in the results were attributed mainly to the enzyme concentrations, since temperature did not cause any variation. A quantitative analysis indicated that the concentration of fructose increased while the concentrations of sucrose and glucose decreased in response to increasing concentrations of the enzymatic complexes. (author)

  5. Enzyme activity and effect of heat treatment on some fungal diseases of postharvest tomato fruits.

    Science.gov (United States)

    Amer, M A; El-Abd, S M; Mansour, F G F

    2013-01-01

    The activity of heat treatments for controlling tomato black mould caused by Alternaria alternata (Fr.) Keissler and grey mould caused by Botrytis cinerea Pers. Ex. Pers. wWas tested. Spore suspension of the grey mold pathogen which was exposed to hot water treatment at 58 degrees C for 6 min., failed to germinate, whereas A. alternata failed to germinate when exposed to hot water at 58 degrees C, for 8 min. Exposing discs bearing growth of any both tested fungi to hot air at 48 degrees C for 72 h caused growth suppression for the both tested pathogens. Dipping tomato fruits in hot water at 58 degrees C for 8 min or keeping in hot air for 72 h at 40 degrees C prevented decay development in non inoculated or artificially inoculated fruits with A. alternata and B. cinerea up to 12 days when stored at 24 degrees C. The high amount of polygalacturonase (PG) and pectinmethylestrase (BME) enzymes were investigated in infected fruits by the two tested pathogens as compared by healthy ones. The culture filtrate of A. alternata had the highest protease activity, white B. cinerea was the lowest. Protease activity was higher when CD liquid medium contain casein was used.

  6. An easily regenerable enzyme reactor prepared from polymerized high internal phase emulsions

    Energy Technology Data Exchange (ETDEWEB)

    Ruan, Guihua, E-mail: guihuaruan@hotmail.com [Guangxi Key Laboratory of Electrochemical and Magnetochemical Functional Materials, College of Chemistry and Bioengineering, Guilin University of Technology, Guangxi 541004 (China); Guangxi Collaborative Innovation Center for Water Pollution Control and Water Safety in Karst Area, Guilin University of Technology, Guilin 541004 (China); Wu, Zhenwei; Huang, Yipeng; Wei, Meiping; Su, Rihui [Guangxi Key Laboratory of Electrochemical and Magnetochemical Functional Materials, College of Chemistry and Bioengineering, Guilin University of Technology, Guangxi 541004 (China); Du, Fuyou, E-mail: dufu2005@126.com [Guangxi Key Laboratory of Electrochemical and Magnetochemical Functional Materials, College of Chemistry and Bioengineering, Guilin University of Technology, Guangxi 541004 (China); Guangxi Collaborative Innovation Center for Water Pollution Control and Water Safety in Karst Area, Guilin University of Technology, Guilin 541004 (China)

    2016-04-22

    A large-scale high-efficient enzyme reactor based on polymerized high internal phase emulsion monolith (polyHIPE) was prepared. First, a porous cross-linked polyHIPE monolith was prepared by in-situ thermal polymerization of a high internal phase emulsion containing styrene, divinylbenzene and polyglutaraldehyde. The enzyme of TPCK-Trypsin was then immobilized on the monolithic polyHIPE. The performance of the resultant enzyme reactor was assessed according to the conversion ability of N{sub α}-benzoyl-L-arginine ethyl ester to N{sub α}-benzoyl-L-arginine, and the protein digestibility of bovine serum albumin (BSA) and cytochrome (Cyt-C). The results showed that the prepared enzyme reactor exhibited high enzyme immobilization efficiency and fast and easy-control protein digestibility. BSA and Cyt-C could be digested in 10 min with sequence coverage of 59% and 78%, respectively. The peptides and residual protein could be easily rinsed out from reactor and the reactor could be regenerated easily with 4 M HCl without any structure destruction. Properties of multiple interconnected chambers with good permeability, fast digestion facility and easily reproducibility indicated that the polyHIPE enzyme reactor was a good selector potentially applied in proteomics and catalysis areas. - Graphical abstract: Schematic illustration of preparation of hypercrosslinking polyHIPE immobilized enzyme reactor for on-column protein digestion. - Highlights: • A reactor was prepared and used for enzyme immobilization and continuous on-column protein digestion. • The new polyHIPE IMER was quite suit for protein digestion with good properties. • On-column digestion revealed that the IMER was easy regenerated by HCl without any structure destruction.

  7. Rapid shifts in Atta cephalotes fungus-garden enzyme activity after a change in fungal substrate (Attini, Formicidae)

    DEFF Research Database (Denmark)

    Kooij, P W; Schiøtt, M; Boomsma, J J

    2011-01-01

    , we measured the changes in enzyme activity after a controlled shift in fungal substrate offered to six laboratory colonies of Atta cephalotes. An ant diet consisting exclusively of grains of parboiled rice rapidly increased the activity of endo-proteinases and some of the pectinases attacking...... from the rice diet, relative to the leaf diet controls. Enzyme activity in the older, bottom sections of fungus gardens decreased, indicating a faster processing of the rice substrate compared to the leaf diet. These results suggest that leaf-cutting ant fungus gardens can rapidly adjust enzyme......Fungus gardens of the basidiomycete Leucocoprinus gongylophorus sustain large colonies of leaf-cutting ants by degrading the plant material collected by the ants. Recent studies have shown that enzyme activity in these gardens is primarily targeted toward starch, proteins and the pectin matrix...

  8. Selection of pectinesterase overproducing fungal strains for the low degree methoxylated pectin preparation.

    Science.gov (United States)

    Aizenberg, V L; Zakharchenko, V A; Semakova, M V; Syrchin, S A; Sedina, S A; Kapichon, A P

    2002-01-01

    The selection of the novel pectinesterase (PE) producing strains is an important step in their biotechnological application. The present work displays the purposeful multistage selection of strains with the increased ability to synthesize extracellular PE among 700 filamentous fungal collection cultures belonging to 45 genera and 93 species. A scheme of targeted multistage screening of PE producers has been developed. At the first stage of the screening 23 potentially active strains belonging to the genera Penicillium, Fusarium, Aspergillus, Rhizopus, Gliocladium, etc., were selected. Strains of the Trichoderma, Cochliobolus, Cladosporium and Thielavia genera not yet described as PE-active ones were selected. The P. funiculosum species (0.28-0.56 units/ml) possessed higher ability to synthesize PE. High level of the PE activity was revealed in P. rubrum (0.37 units/ml). No correlation between the level of fungal biomass accumulation on pectin-containing medium and the level of their PE activity was observed. A principal chance of the fungal PE substitution for some chemical reagents at the stage of demethoxylation of the pectin-containing raw materials should be noted. The technology of the fungal PE application for production of the low-degree methoxylated pectin is under development. Such pectin is necessary for the low-calorie jellfying products in the food industry and in medicine as a preventive agent removing the toxic substances from the human body.

  9. Screening of Argentine native fungal strains for biocontrol of the grasshopper Tropidacris collaris: relationship between fungal pathogenicity and chitinolytic enzyme activity.

    Science.gov (United States)

    Pelizza, S A; Elíades, L A; Saparrat, M C N; Cabello, M N; Scorsetti, A C; Lange, C E

    2012-04-01

    Tropidacris collaris (Orthoptera: Romaleidae) is a large and voracious grasshopper, which, in recent years, has become a recurrent pest in increasingly extensive areas of Argentina's northern provinces. In the present work chitinase activity was measured in 59 entomopathogenic fungal isolates native to Argentina, and the relationship between enzymatic activity and fungal virulence was assessed. Isolate LPSC 1067 caused the highest mortality on T. collaris nymphs (97.7 ± 1.22%). Nine isolates caused no mortality, while the remaining 49 caused mortalities ranging from 6.6 ± 0.3% (LPSC 770) to 91.06 ± 1.51% (LPSC 906). Several isolates revealed chitinolytic capabilities on test plates, although the activities differed with respect to the ratio of the chitin-decay-halo and fungal-colony diameters. A principal component analysis indicated that isolate LPSC 1067, obtained from a long-horned grasshopper (Orthoptera: Tettigoniidae), would be a potential candidate for T. collaris biocontrol because the strain exhibited the highest mortality, a shorter median lethal time, and a high enzymatic activity and growth rate.

  10. Enzyme production in immobilized Trichoderma reesei cells with hydrophobic polymers prepared by radiation polymerization method

    International Nuclear Information System (INIS)

    Luzhao Xin; Kumakura, Minoru; Kaetsu, Isao

    1993-01-01

    Trichoderma reesei cells were immobilized on paper covered with hydrophobic monomer, trimethylpropane triacrylate by radiation polymerization. The effect of immobilization condition on enzyme productivity was studied by measuring filter paper and cellobiose activity. The cells were adhered and grew on the surface of the carrier with the polymer giving high enzyme productivity in the immobilized cells in comparison with the free cells. Optimum concentration and volume of the coating monomer for the preparation of the immobilized cells were obtained. (author)

  11. Bitterness in sodium caseinate hydrolysates: role of enzyme preparation and degree of hydrolysis.

    Science.gov (United States)

    O'Sullivan, Dara; Nongonierma, Alice B; FitzGerald, Richard J

    2017-10-01

    Enzymatic hydrolysis of sodium caseinate (NaCas) may lead to the development of bitterness. Careful selection of hydrolysis conditions (i.e. enzyme preparation and duration) yielding different degrees of hydrolysis (DH) may aid in the development of low bitterness. Eighteen NaCas hydrolysates were generated with four enzyme preparations (Alcalase 2.4L, Prolyve 1000, FlavorPro Whey and pepsin) to different DH values. Hydrolysate bitterness score, assessed using a trained panel (ten assessors), generally increased at higher DH values for Alcalase, Prolyve and pepsin hydrolysates. However, all FlavorPro Whey hydrolysates (DH 0.38-10.62%) displayed low bitterness score values ( 0.05). Enzyme preparation and DH affect the bitterness of NaCas hydrolysates. The results are relevant for the generation of NaCas hydrolysates with reduced bitterness. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  12. Enzyme

    Science.gov (United States)

    Enzymes are complex proteins that cause a specific chemical change in all parts of the body. For ... use them. Blood clotting is another example of enzymes at work. Enzymes are needed for all body ...

  13. Changes in bacterial and fungal communities across compost recipes, preparation methods, and composting times.

    Science.gov (United States)

    Neher, Deborah A; Weicht, Thomas R; Bates, Scott T; Leff, Jonathan W; Fierer, Noah

    2013-01-01

    Compost production is a critical component of organic waste handling, and compost applications to soil are increasingly important to crop production. However, we know surprisingly little about the microbial communities involved in the composting process and the factors shaping compost microbial dynamics. Here, we used high-throughput sequencing approaches to assess the diversity and composition of both bacterial and fungal communities in compost produced at a commercial-scale. Bacterial and fungal communities responded to both compost recipe and composting method. Specifically, bacterial communities in manure and hay recipes contained greater relative abundances of Firmicutes than hardwood recipes with hay recipes containing relatively more Actinobacteria and Gemmatimonadetes. In contrast, hardwood recipes contained a large relative abundance of Acidobacteria and Chloroflexi. Fungal communities of compost from a mixture of dairy manure and silage-based bedding were distinguished by a greater relative abundance of Pezizomycetes and Microascales. Hay recipes uniquely contained abundant Epicoccum, Thermomyces, Eurotium, Arthrobotrys, and Myriococcum. Hardwood recipes contained relatively abundant Sordariomycetes. Holding recipe constant, there were significantly different bacterial and fungal communities when the composting process was managed by windrow, aerated static pile, or vermicompost. Temporal dynamics of the composting process followed known patterns of degradative succession in herbivore manure. The initial community was dominated by Phycomycetes, followed by Ascomycota and finally Basidiomycota. Zygomycota were associated more with manure-silage and hay than hardwood composts. Most commercial composters focus on the thermophilic phase as an economic means to insure sanitation of compost from pathogens. However, the community succeeding the thermophilic phase begs further investigation to determine how the microbial dynamics observed here can be best managed

  14. Changes in bacterial and fungal communities across compost recipes, preparation methods, and composting times.

    Directory of Open Access Journals (Sweden)

    Deborah A Neher

    Full Text Available Compost production is a critical component of organic waste handling, and compost applications to soil are increasingly important to crop production. However, we know surprisingly little about the microbial communities involved in the composting process and the factors shaping compost microbial dynamics. Here, we used high-throughput sequencing approaches to assess the diversity and composition of both bacterial and fungal communities in compost produced at a commercial-scale. Bacterial and fungal communities responded to both compost recipe and composting method. Specifically, bacterial communities in manure and hay recipes contained greater relative abundances of Firmicutes than hardwood recipes with hay recipes containing relatively more Actinobacteria and Gemmatimonadetes. In contrast, hardwood recipes contained a large relative abundance of Acidobacteria and Chloroflexi. Fungal communities of compost from a mixture of dairy manure and silage-based bedding were distinguished by a greater relative abundance of Pezizomycetes and Microascales. Hay recipes uniquely contained abundant Epicoccum, Thermomyces, Eurotium, Arthrobotrys, and Myriococcum. Hardwood recipes contained relatively abundant Sordariomycetes. Holding recipe constant, there were significantly different bacterial and fungal communities when the composting process was managed by windrow, aerated static pile, or vermicompost. Temporal dynamics of the composting process followed known patterns of degradative succession in herbivore manure. The initial community was dominated by Phycomycetes, followed by Ascomycota and finally Basidiomycota. Zygomycota were associated more with manure-silage and hay than hardwood composts. Most commercial composters focus on the thermophilic phase as an economic means to insure sanitation of compost from pathogens. However, the community succeeding the thermophilic phase begs further investigation to determine how the microbial dynamics observed here

  15. Biodegradation of paraffin wax by crude Aspergillus enzyme preparations for potential use in removing paraffin deposits.

    Science.gov (United States)

    Zhang, Junhui; Xue, Quanhong; Gao, Hui; Wang, Ping

    2015-11-01

    Paraffin deposition problems have plagued the oil industry. Whist mechanical and chemical methods are problematic, microbiological method of paraffin removal is considered an alternative. However, studies have mainly investigated the use of bacteria, with little attention to the potential of fungi. The performance of six Aspergillus isolates to degrade paraffin wax was evaluated under laboratory conditions using solid enzyme preparations. The results showed that all the six enzyme preparations efficiently improved the solubility of paraffin wax in n-hexane and degraded n-alkanes in paraffin wax. The degradation process was accompanied by dynamic production of gases (CO2 and H2 ) and organic acids (oxalate and propionate). The shape of wax crystals markedly changed after enzymatic degradation, with a rough surface and a loose structure. This study indicates that extracellular enzymes from Aspergillus spp. can efficiently degrade paraffin wax. These enzyme preparations have the potential for use in oil wells with paraffin deposition problems. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Enrichment and Broad Representation of Plant Biomass-Degrading Enzymes in the Specialized Hyphal Swellings of Leucoagaricus gongylophorus, the Fungal Symbiont of Leaf-Cutter Ants

    Energy Technology Data Exchange (ETDEWEB)

    Aylward, Frank O.; Khadempour, Lily; Tremmel, Daniel; McDonald, Bradon R.; Nicora, Carrie D.; Wu, Si; Moore, Ronald J.; Orton, Daniel J.; Monroe, Matthew E.; Piehowski, Paul D.; Purvine, Samuel O.; Smith, Richard D.; Lipton, Mary S.; Burnum-Johnson, Kristin E.; Currie, Cameron R.

    2015-08-28

    Leaf-cutter ants are prolific and conspicuous Neotropical herbivores that derive energy from specialized fungus gardens they cultivate using foliar biomass. The basidiomycetous cultivar of the ants, Leucoagaricus gongylophorus, produces specialized hyphal swellings called gongylidia that serve as the primary food source of ant colonies. Gongylidia also contain lignocellulases that become concentrated in ant digestive tracts and are deposited within fecal droplets onto fresh foliar material as it is foraged by the ants. Although the enzymes concentrated by L. gongylophorus within gongylidia are thought to be critical to the initial degradation of plant biomass, only a few enzymes present in these hyphal swellings have been identified. Here we use proteomic methods to identify proteins present in the gongylidia of three Atta cephalotes colonies. Our results demonstrate that a diverse but consistent set of enzymes is present in gongylidia, including numerous lignocellulases likely involved in the degradation of polysaccharides, plant toxins, and proteins. Overall, gongylidia contained over three-quarters of all lignocellulases identified in the L. gongylophorus genome, demonstrating that the majority of the enzymes produced by this fungus for biomass breakdown are ingested by the ants. We also identify a set of 23 lignocellulases enriched in gongylidia compared to whole fungus garden samples, suggesting that certain enzymes may be particularly important in the initial degradation of foliar material. Our work sheds light on the complex interplay between leaf-cutter ants and their fungal symbiont that allows for the host insects to occupy an herbivorous niche by indirectly deriving energy from plant biomass.

  17. Redistribution of mineral elements in wheat grain when applying the complex enzyme preparations based on phytase

    Directory of Open Access Journals (Sweden)

    Elena Kuznetsova

    2016-01-01

    Full Text Available Biogenic minerals play an important role in the whole human nutrition, but they are included in the grain of the phytates that reduces their bioavailability. Whole wheat bread is generally considered a healthy food, but the presence of mineral elements in it is insignificant, because of weak phytate degradation. From all sources of exogenous phytase the most productive are microscopic fungi. To accelerate the process of transition hard mineral elements are mobilized to implement integrated cellulolytic enzyme preparation based on the actions of phytase (producer is Penicillium canescens. Phytase activity was assessed indirectly by the rate of release of phosphate from the substrate. It has been established that the release rate of the phosphoric acid substrate is dependent on the composition of the drug and the enzyme complex is determined by the presence of xylanase. The presented experimental data shows that a cellulase treatment of the grain in conjunction with the β-glucanase or xylanase leading to an increase in phytase activity could be 1.4 - 2.3 times as compared with the individual enzymes. As a result of concerted action of enzymes complex preparation varies topography grain, increase the pore sizes in seed and fruit shells that facilitate the penetration of the enzyme phytase in the aleurone layer to the site of phytin hydrolysis and leads to an increase in phytase activity. In terms of rational parameters of enzymatic hydrolysis, the distribution of mineral elements in the anatomical parts of the grain after processing complex enzyme preparation with the help of X-ray detector EMF miniCup system in a scanning electron microscope JEOL JSM 6390 were investigated. When processing enzyme preparation wheat trend in the distribution of mineral elements, characteristic of grain - the proportion of these elements in the aleurone layer decreases, and in the endosperm increases. Because dietary fiber and phytate found together in the

  18. Stable preparations of tyrosine hydroxylase provide the solution structure of the full-length enzyme

    Science.gov (United States)

    Bezem, Maria T.; Baumann, Anne; Skjærven, Lars; Meyer, Romain; Kursula, Petri; Martinez, Aurora; Flydal, Marte I.

    2016-01-01

    Tyrosine hydroxylase (TH) catalyzes the rate-limiting step in the biosynthesis of catecholamine neurotransmitters. TH is a highly complex enzyme at mechanistic, structural, and regulatory levels, and the preparation of kinetically and conformationally stable enzyme for structural characterization has been challenging. Here, we report on improved protocols for purification of recombinant human TH isoform 1 (TH1), which provide large amounts of pure, stable, active TH1 with an intact N-terminus. TH1 purified through fusion with a His-tagged maltose-binding protein on amylose resin was representative of the iron-bound functional enzyme, showing high activity and stabilization by the natural feedback inhibitor dopamine. TH1 purified through fusion with a His-tagged ZZ domain on TALON is remarkably stable, as it was partially inhibited by resin-derived cobalt. This more stable enzyme preparation provided high-quality small-angle X-ray scattering (SAXS) data and reliable structural models of full-length tetrameric TH1. The SAXS-derived model reveals an elongated conformation (Dmax = 20 nm) for TH1, different arrangement of the catalytic domains compared with the crystal structure of truncated forms, and an N-terminal region with an unstructured tail that hosts the phosphorylation sites and a separated Ala-rich helical motif that may have a role in regulation of TH by interacting with binding partners. PMID:27462005

  19. Rapid preparation of functional polysaccharides from Pyropia yezoensis by microwave-assistant rapid enzyme digest system.

    Science.gov (United States)

    Lee, Ji-Hyeok; Kim, Hyung-Ho; Ko, Ju-Young; Jang, Jun-Ho; Kim, Gwang-Hoon; Lee, Jung-Suck; Nah, Jae-Woon; Jeon, You-Jin

    2016-11-20

    This study describes a simple preparation of functional polysaccharides from Pyropia yezoensis using a microwave-assistant rapid enzyme digest system (MAREDS) with various carbohydrases, and evaluates their antioxidative effects. Polysaccharide hydrolysates were prepared using MAREDS under different hydrolytic conditions of the carbohydrases and microwave powers. Polysaccharides less than 10kDa (Low molecular weight polysaccharides, LMWP, ≤10kDa) were efficiently obtained using an ultrafiltration (molecular weight cut-off of 10kDa). MAREDS increases AMG activation via an increased degree of hydrolysis; the best AMG hydrolysate was prepared using a 10:1 ratio of substrate to enzyme for 2h in MAREDS with 400W. LMWP consisted of galactose (27.3%), glucose (64.5%), and mannose (8.3%) from the AMG hydrolysate had stronger antioxidant effects than the high molecular weight polysaccharides (>10kDa). We rapidly prepared functional LMWPs by using MAREDS with carbohydrases, and suggest that LMWP might be potentially a valuable algal polysaccharide antioxidant. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Preparation and Optimisation of Cross-Linked Enzyme Aggregates Using Native Isolate White Rot Fungi Trametes versicolor and Fomes fomentarius for the Decolourisation of Synthetic Dyes

    Science.gov (United States)

    Vršanská, Martina; Voběrková, Stanislava; Jiménez Jiménez, Ana María; Strmiska, Vladislav; Adam, Vojtěch

    2017-01-01

    The key to obtaining an optimum performance of an enzyme is often a question of devising a suitable enzyme and optimisation of conditions for its immobilization. In this study, laccases from the native isolates of white rot fungi Fomes fomentarius and/or Trametes versicolor, obtained from Czech forests, were used. From these, cross-linked enzyme aggregates (CLEA) were prepared and characterised when the experimental conditions were optimized. Based on the optimization steps, saturated ammonium sulphate solution (75 wt.%) was used as the precipitating agent, and different concentrations of glutaraldehyde as a cross-linking agent were investigated. CLEA aggregates formed under the optimal conditions showed higher catalytic efficiency and stabilities (thermal, pH, and storage, against denaturation) as well as high reusability compared to free laccase for both fungal strains. The best concentration of glutaraldehyde seemed to be 50 mM and higher efficiency of cross-linking was observed at a low temperature 4 °C. An insignificant increase in optimum pH for CLEA laccases with respect to free laccases for both fungi was observed. The results show that the optimum temperature for both free laccase and CLEA laccase was 35 °C for T. versicolor and 30 °C for F. fomentarius. The CLEAs retained 80% of their initial activity for Trametes and 74% for Fomes after 70 days of cultivation. Prepared cross-linked enzyme aggregates were also investigated for their decolourisation activity on malachite green, bromothymol blue, and methyl red dyes. Immobilised CLEA laccase from Trametes versicolor showed 95% decolourisation potential and CLEA from Fomes fomentarius demonstrated 90% decolourisation efficiency within 10 h for all dyes used. These results suggest that these CLEAs have promising potential in dye decolourisation. PMID:29295505

  1. Olive mill wastewater biodegradation potential of white-rot fungi--Mode of action of fungal culture extracts and effects of ligninolytic enzymes.

    Science.gov (United States)

    Ntougias, Spyridon; Baldrian, Petr; Ehaliotis, Constantinos; Nerud, Frantisek; Merhautová, Věra; Zervakis, Georgios I

    2015-01-01

    Forty-nine white-rot strains belonging to 38 species of Basidiomycota were evaluated for olive-mill wastewater (OMW) degradation. Almost all fungi caused high total phenolics (>60%) and color (⩽ 70%) reduction, while COD and phytotoxicity decreased to a lesser extent. Culture extracts from selected Agrocybe cylindracea, Inonotus andersonii, Pleurotus ostreatus and Trametes versicolor strains showed non-altered physicochemical and enzymatic activity profiles when applied to raw OMW in the presence or absence of commercial catalase, indicating no interaction of the latter with fungal enzymes and no competition for H2O2. Hydrogen peroxide's addition resulted in drastic OMW's decolorization, with no effect on phenolic content, suggesting that oxidation affects colored components, but not necessarily phenolics. When fungal extracts were heat-treated, no phenolics decrease was observed demonstrating thus their enzymatic rather than physicochemical oxidation. Laccases added to OMW were reversibly inhibited by the effluent's high phenolic load, while peroxidases were stable and active during the entire process. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. The use of combined radiation methods for decreasing the bacterial dissemination of enzyme preparations

    International Nuclear Information System (INIS)

    Samojlenko, I.I.; Fedotov, N.S.; Tumanyan, M.A.; Korolev, N.I.

    1984-01-01

    A study was made on possibility of using ionizing radiation in combination with alternative magnetic field (AMF) and heating for decreasing the bacterial dissemination of proteolytic enzymes. Papain, trypsin, chymotrypsin and amylorysin (the preparation possessing proteolytic and amylolytic activities) were subjected to gamma irradiation at 10-25 kGy dose range, the effect of AMF with 750 oe and heating at 50 deg during 60 min. Model tests conducted with the use of Escherichia Coli cells and Bacillus anthracoides spores showed that survival rate of bacteria irradiated in protective medium was lower in the case of combined magnetoradiation and thermoradiation effect. The use of 10 kGy dose of ionizing radiation in combination with treatment in alternative magnetic field or with heating provided the required decrease of dissemination of irradiated enzyme samples with complete conservation of proteolytic activity by them

  3. Modulation of liver enzymes by an Iranian preparation of Echinacea purpurea

    Directory of Open Access Journals (Sweden)

    A. Manayi

    2015-10-01

    Full Text Available Hepatitis B, a common infectious disease of liver, is transmitted by blood and body fluids like semen and vaginal fluid that carry hepatitis B virus (HBV.  In chronic infection, medical care is required to decrease possibility of cirrhosis and liver cancer. In the present report, the hepatoprotective effect of an Echinacea purpurea preparation (Echiherb® has been described in a patient who suffered from HBV infection. The levels of both enzymes of aspartate aminotransferase (AST and alanine aminotransferase (ALT decreased to their normal level after 6 weeks of treatment. Therefore, this report may provide a new perspective for protection of liver in patients with HBV infection along with other diseases which damage liver cells using E. purpurea preparations.

  4. Antitumor activity of a fungal glucan tylopilan and Propionibacterium acnes preparation

    Directory of Open Access Journals (Sweden)

    Jan Grzybek

    2014-01-01

    Full Text Available The study evaluated the antitumor activity of tylopilan, aβ- (1→3 (1→6 linked glucan isolated from fruiting bodies of Tylopilus felleus (Bull.: Fr. P. Karst. (Boletaceae, and Propionibacterium acnes (P.a. preparation. The antitumor effect of tylopilan and P.a. used alone or in combination was studied in NMRI mice inoculated i.p. with 106 180-TG Crocker tumor cells. All experiments were based on a pretreatment with tylopilan and/or P.a. 5 days and/or 2 h before tumor cell inoculation. Mean survival time (MST of tumor - bearing mice was significantly prolonged in comparison to control mice by a single injection of tylopilan (25 µg/mouse or 50 µg/mouse or P.a. (1 mg/mouse. MST was 23.6; 22.8 days in the tylopilan injected mice and 17.5 in the control animals. Tylopilan injected in conjunction with P.a. prolonged signifi-cantly MST in comparison to control mice as well as to tylopilan alone treated mice. We have found that P.a. which stimulate immune response enhanced significantly antitumor activity of tylopilan. The cytotoxicity of tylopilan at concentrations of 300, 150, 75 and 37.5 µg/ml towards 180-TG Crocker cells in vitro studies was evaluated. All examined tylopilan concentrations showed cytotoxic activity.

  5. On-site hydrolytic enzymes production from fungal co-cultivation of Bermuda grass and corn cob.

    Science.gov (United States)

    Amaro-Reyes, Aldo; Gracida, Jorge; Huizache-Peña, Nelson; Elizondo-García, Norberto; Salazar-Martínez, José; García Almendárez, Blanca E; Regalado, Carlos

    2016-07-01

    Solid state fermentation (SSF) is used to produce industrial enzymes. The objective of this study was to use a co-culture of Aspergillus niger GS1 and Trichoderma reesei, grown on a mixture of Bermuda grass and corn cob to obtain fermented forage (FF) rich in hydrolytic enzymes, as a value added ingredient for animal feed. FPase, amylase and xylanase productivities (dry matter, DM) were 8.8, 181.4, and 42.1Ug(-1)h(-1), respectively (1U=reducing sugars released min(-1)), after 12-16h of SSF with C/N=60. Cellulose, hemicellulose and lignin decreased 1.6-, 2.7- and 1.9-fold (DM), respectively. In vitro ruminal and true digestibility of DM was improved 2.4- and 1.4-fold. Ruminal digestion of FF reduced 1.32-fold the acetate:propionate ratio, which may reduce the environmental impact of ruminants feeding. On-site hydrolytic enzymes productivity using SSF without enzymes extraction could be of economic potential for digestibility improvement in animal feed. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Development and selection of fungal and bacterial mutants using ionizing radiation and radioisotopes for improved enzyme production (cellulase and coagulase)

    International Nuclear Information System (INIS)

    Markov, K.I.

    1975-01-01

    Ultraviolet and gamma radiations, chemical mutagens, and combinations of chemical and physical mutagens were used in order to obtain mutants of Bacillus mesentericus and Trichoderma viridae with a higher production of coagulase and cellulase, respectively. It was possible to isolate mutant strains, with enzyme activity increased by a factor of 2 and 3

  7. Facile Method To Prepare Microcapsules Inspired by Polyphenol Chemistry for Efficient Enzyme Immobilization.

    Science.gov (United States)

    Zhang, Shaohua; Jiang, Zhongyi; Wang, Xiaoli; Yang, Chen; Shi, Jiafu

    2015-09-09

    In this study, a method inspired by polyphenol chemistry is developed for the facile preparation of microcapsules under mild conditions. Specifically, the preparation process includes four steps: formation of the sacrificial template, generation of the polyphenol coating on the template surface, cross-linking of the polyphenol coating by cationic polymers, and removal of the template. Tannic acid (TA) is chosen as a representative polyphenol coating precursor for the preparation of microcapsules. The strong interfacial affinity of TA contributes to the formation of polyphenol coating through oxidative oligomerization, while the high reactivity of TA is in charge of reacting/cross-linking with cationic polymer polyethylenimine (PEI) through Schiff base/Michael addition reaction. The chemical/topological structures of the resultant microcapsules are simultaneously characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier Transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), etc. The wall thickness of the microcapsules could be tailored from 257±20 nm to 486±46 nm through changing the TA concentration. The microcapsules are then utilized for encapsulating glucose oxidase (GOD), and the immobilized enzyme exhibits desired catalytic activity and enhanced pH and thermal stabilities. Owing to the structural diversity and functional versatility of polyphenols, this study may offer a facile and generic method to prepare microcapsules and other kinds of functional porous materials.

  8. Do new cellulolytic enzyme preparations affect the industrial strategies for high solids lignocellulosic ethanol production?

    Science.gov (United States)

    Cannella, David; Jørgensen, Henning

    2014-01-01

    Production of ethanol from lignocellulosic materials has a promising market potential, but the process is still only at pilot/demonstration scale due to the technical and economical difficulties of the process. Operating the process at very high solids concentrations (above 20% dry matter-DM) has proven essential for economic feasibility at industrial scale. Historically, simultaneous saccharification and fermentation (SSF) was found to give better ethanol yields compared to separate hydrolysis and fermentation (SHF), but data in literature are typically based on operating the process at low dry matter conditions. In this work the impact of selected enzyme preparation and processing strategy (SHF, presaccharification and simultaneous saccharification and fermentation-PSSF, and SSF) on final ethanol yield and overall performance was investigated with pretreated wheat straw up to 30% DM. The experiments revealed that an SSF strategy was indeed better than SHF when applying an older generation enzyme cocktail (Celluclast-Novozym 188). In case of the newer product Cellic CTec 2, SHF resulted in 20% higher final ethanol yield compared to SSF. It was possible to close the mass balance around cellulose to around 94%, revealing that the most relevant products could be accounted for. One observation was the presence of oxidized sugar (gluconic acid) upon enzymatic hydrolysis with the latest enzyme preparation. Experiments showed gluconic acid formation by recently discovered enzymatic class of lytic polysaccharides monoxygenases (LPMO's) to be depending on the processing strategy. The lowest concentration was achieved in SSF, which could be correlated with less available oxygen due to simultaneous oxygen consumption by the yeast. Quantity of glycerol and cell mass was also depending on the selected processing strategy. © 2013 Wiley Periodicals, Inc.

  9. Role of hydrogen peroxide and antioxidant enzymes in the interaction between a hemibiotrophic fungal pathogen, Leptosphaeria maculans, and oilseed rape

    Czech Academy of Sciences Publication Activity Database

    Jindřichová, Barbora; Fodor, J.; Šindelářová, Milada; Burketová, Lenka; Valentová, O.

    2011-01-01

    Roč. 72, č. 2 (2011), s. 149-156 ISSN 0098-8472 R&D Projects: GA ČR GA522/08/1581; GA MŠk MEB040923 Institutional research plan: CEZ:AV0Z50380511 Keywords : hydrogen peroxide * antioxidant enzymes * hemibiotrophic pathogen Subject RIV: GF - Plant Pathology, Vermin, Weed, Plant Protection Impact factor: 2.985, year: 2011

  10. Performance of hemicellulolytic enzymes in culture supernatants from a wide range of fungi on insoluble wheat straw and corn fiber fractions

    NARCIS (Netherlands)

    Gool, van M.P.; Toth, K.; Schols, H.A.; Szakacs, G.; Gruppen, H.

    2012-01-01

    Filamentous fungi are a good source of hemicellulolytic enzymes for biomass degradation. Enzyme preparations were obtained as culture supernatants from 78 fungal isolates grown on wheat straw as carbon source. These enzyme preparations were utilized in the hydrolysis of insoluble wheat straw and

  11. 21 CFR 173.115 - Alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation derived from a recombinant Bacillus...

    Science.gov (United States)

    2010-04-01

    ... preparation derived from a recombinant Bacillus subtilis. 173.115 Section 173.115 Food and Drugs FOOD AND DRUG... Bacillus subtilis. The food additive alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation, may be... derived from a modified Bacillus subtilis strain that contains the gene coding for α-ALDC from Bacillus...

  12. Expression of fungal cutinase and swollenin in tobacco chloroplasts reveals novel enzyme functions and/or substrates.

    Directory of Open Access Journals (Sweden)

    Dheeraj Verma

    Full Text Available In order to produce low-cost biomass hydrolyzing enzymes, transplastomic lines were generated that expressed cutinase or swollenin within chloroplasts. While swollenin expressing plants were homoplasmic, cutinase transplastomic lines remained heteroplasmic. Both transplastomic lines showed interesting modifications in their phenotype, chloroplast structure, and functions. Ultrastructural analysis of chloroplasts from cutinase- and swollenin-expressing plants did not show typical lens shape and granal stacks. But, their thylakoid membranes showed unique scroll like structures and chloroplast envelope displayed protrusions, stretching into the cytoplasm. Unusual honeycomb structures typically observed in etioplasts were observed in mature chloroplasts expressing swollenin. Treatment of cotton fiber with chloroplast-derived swollenin showed enlarged segments and the intertwined inner fibers were irreversibly unwound and fully opened up due to expansin activity of swollenin, causing disruption of hydrogen bonds in cellulose fibers. Cutinase transplastomic plants showed esterase and lipase activity, while swollenin transplastomic lines lacked such enzyme activities. Higher plants contain two major galactolipids, monogalactosyldiacylglycerol (MGDG and digalactosyldiacylglycerol (DGDG, in their chloroplast thylakoid membranes that play distinct roles in their structural organization. Surprisingly, purified cutinase effectively hydrolyzed DGDG to MGDG, showing alpha galactosidase activity. Such hydrolysis resulted in unstacking of granal thylakoids in chloroplasts and other structural changes. These results demonstrate DGDG as novel substrate and function for cutinase. Both MGDG and DGDG were reduced up to 47.7% and 39.7% in cutinase and 68.5% and 67.5% in swollenin expressing plants. Novel properties and functions of both enzymes reported here for the first time should lead to better understanding and enhanced biomass hydrolysis.

  13. Fungal Keratitis

    Science.gov (United States)

    ... Español Eye Health / Eye Health A-Z Fungal Keratitis Sections What is Fungal Keratitis? Fungal Keratitis Causes ... Keratitis Symptoms Fungal Keratitis Treatment What is Fungal Keratitis? Leer en Español: ¿Qué Es la Queratitis Fúngica? ...

  14. Assessment of Dextran Antigenicity of Intravenous Iron Preparations with Enzyme-Linked Immunosorbent Assay (ELISA

    Directory of Open Access Journals (Sweden)

    Susann Neiser

    2016-07-01

    Full Text Available Intravenous iron preparations are typically classified as non-dextran-based or dextran/dextran-based complexes. The carbohydrate shell for each of these preparations is unique and is key in determining the various physicochemical properties, the metabolic pathway, and the immunogenicity of the iron-carbohydrate complex. As intravenous dextran can cause severe, antibody-mediated dextran-induced anaphylactic reactions (DIAR, the purpose of this study was to explore the potential of various intravenous iron preparations, non-dextran-based or dextran/dextran-based, to induce these reactions. An IgG-isotype mouse monoclonal anti-dextran antibody (5E7H3 and an enzyme-linked immunosorbent assay (ELISA were developed to investigate the dextran antigenicity of low molecular weight iron dextran, ferumoxytol, iron isomaltoside 1000, ferric gluconate, iron sucrose and ferric carboxymaltose, as well as isomaltoside 1000, the isolated carbohydrate component of iron isomaltoside 1000. Low molecular weight iron dextran, as well as dextran-based ferumoxytol and iron isomaltoside 1000, reacted with 5E7H3, whereas ferric carboxymaltose, iron sucrose, sodium ferric gluconate, and isolated isomaltoside 1000 did not. Consistent results were obtained with reverse single radial immunodiffusion assay. The results strongly support the hypothesis that, while the carbohydrate alone (isomaltoside 1000 does not form immune complexes with anti-dextran antibodies, iron isomaltoside 1000 complex reacts with anti-dextran antibodies by forming multivalent immune complexes. Moreover, non-dextran based preparations, such as iron sucrose and ferric carboxymaltose, do not react with anti-dextran antibodies. This assay allows to assess the theoretical possibility of a substance to induce antibody-mediated DIARs. Nevertheless, as this is only one possible mechanism that may cause a hypersensitivity reaction, a broader set of assays will be required to get an understanding of the

  15. Aged-look vat dyed cotton with anti-bacterial/anti-fungal properties by treatment with nano clay and enzymes.

    Science.gov (United States)

    Maryan, Ali Sadeghian; Montazer, Majid; Harifi, Tina; Rad, Mahnaz Mahmoudi

    2013-06-05

    In this research, nanotechnology as a route to functional finishing of textiles was used along with bio-finishing to enhance the cotton fabrics performance. For this purpose, quaternary modified montmorillonite and common enzymes such as cellulase, laccase and their mixture were applied on vat dyed cotton fabric. Characteristic analysis of the treated samples and the dispersed nano clays in the effluent of the treatment was performed by various analyzing methods. The nano/bio-finishing is believed to impart antibacterial and antifungal activities with simultaneously higher lightness, advanced softness and handle properties into cotton fabrics. Moreover, cotton fabrics were proved to have no adverse effects (low toxicity) on human dermal fibroblasts. Findings suggest the potential of the proposed method in reducing the risk of microorganism for textile applications and imparting better handle and appearance properties. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Comparing the physiochemical parameters of three celluloses reveals new insights into substrate suitability for fungal enzyme production.

    Science.gov (United States)

    Hassan, Lara; Reppke, Manfred J; Thieme, Nils; Schweizer, Steffen A; Mueller, Carsten W; Benz, J Philipp

    2017-01-01

    The industrial applications of cellulases are mostly limited by the costs associated with their production. Optimized production pathways are therefore desirable. Based on their enzyme inducing capacity, celluloses are commonly used in fermentation media. However, the influence of their physiochemical characteristics on the production process is not well understood. In this study, we examined how physical, structural and chemical properties of celluloses influence cellulase and hemicellulase production in an industrially-optimized and a non-engineered filamentous fungus: Trichoderma reesei RUT-C30 and Neurospora crassa . The performance was evaluated by quantifying gene induction, protein secretion and enzymatic activities. Among the three investigated substrates, the powdered cellulose was found to be the most impure, and the residual hemicellulosic content was efficiently perceived by the fungi. It was furthermore found to be the least crystalline substrate and consequently was the most readily digested cellulose in vitro. In vivo however, only RUT-C30 was able to take full advantage of these factors. When comparing carbon catabolite repressed and de-repressed strains of T. reesei and N. crassa , we found that cre1 / cre - 1 is at least partially responsible for this observation, but that the different wiring of the molecular signaling networks is also relevant. Our findings indicate that crystallinity and hemicellulose content are major determinants of performance. Moreover, the genetic background between WT and modified strains greatly affects the ability to utilize the cellulosic substrate. By highlighting key factors to consider when choosing the optimal cellulosic product for enzyme production, this study has relevance for the optimization of a critical step in the biotechnological (hemi-) cellulase production process.

  17. Enzyme and fungal treatments and a combination thereof reduce olive mill wastewater phytotoxicity on Zea mays L. seeds.

    Science.gov (United States)

    Quaratino, Daniele; D'Annibale, Alessandro; Federici, Federico; Cereti, Carlo Fausto; Rossini, Francesco; Fenice, Massimiliano

    2007-01-01

    The phytotoxicity of olive-mill wastewater (OMW) has been suggested to be mainly due to its phenolic components. This study investigated the impact of three different low-cost dephenolization treatments on the wastewater phytotoxicity. To this aim, germinability of maize (Zea mays L.) seeds sown on a sandy-loamy soil which had been spread with different volumes (from 40 to 160m(3)ha(-1)) of either biologically-treated OMW or relative incubation control was determined. Biological treatments included either Panus tigrinus liquid cultures or incubation with commercial laccase (1UIml(-1)) or an innovative sequential combination of laccase and P. tigrinus cultures. All treatments markedly reduced phytotoxicity and promising results were obtained with commercial laccase. In fact, germinability and mean germination times in soil spread with laccase-treated OMW, did not significantly differ from those observed in soil irrigated with tap water (control) up to OMW volumes of 120m(3)ha(-1). Although the highest phenol reduction (ca. 81%) was obtained by the sequential use of laccase and P. tigrinus, the feasibility of the enzyme treatment is undoubtedly more convincing under the technological point of view.

  18. β-(1 → 3)-Glucanolytic yeasts from Brazilian grape microbiota: production and characterization of β-glucanolytic enzymes by Aureobasidium pullulans 1WA1 cultivated on fungal Mycelium.

    Science.gov (United States)

    Bauermeister, Anelize; Amador, Ismael R; Pretti, Carla P; Giese, Ellen C; Oliveira, André L M

    2015-01-14

    A total of 95 yeast strains were isolated from the microbiota of different grapes collected at vineyards in southern Brazil. The yeasts were screened for β-(1 → 3)-glucanases using a newly developed zymogram method that relies upon the appearance of clearance zones around growing colonies cultured on agar–botryosphaeran medium and also by submerged fermentation on nutrient medium containing botryosphaeran, a (1 → 3),(1 → 6)-β-d-glucan. Among 14 β-(1 → 3)-glucanase-positive yeasts identified, four strains produced the highest β-glucanolytic activities and were evaluated for enzyme production on cellobiose, botryosphaeran, and mycelial biomass from Botryosphaeria rhodina (MAMB-05). Yeast strain 1WA1 produced the highest β-(1 → 3)-glucanase and β-glucosidase activities and was identified by molecular characterization as Aureobasidium pullulans. The physicochemical properties of the crude β-glucanolytic enzyme preparation were characterized, and the preparation was used to hydrolyze several β-d-glucans (laminarin, botryosphaeran, lasiodiplodan, pustulan, and curdlan). The production and physicochemical properties of the β-glucanolytic preparation enable its potential applications in wine enology and production of prebiotics through hydrolysis of β-d-glucans.

  19. Carbonic anhydrase inhibitors: inhibition of the beta-class enzymes from the fungal pathogens Candida albicans and Cryptococcus neoformans with simple anions.

    Science.gov (United States)

    Innocenti, Alessio; Mühlschlegel, Fritz A; Hall, Rebecca A; Steegborn, Clemens; Scozzafava, Andrea; Supuran, Claudiu T

    2008-09-15

    The catalytic activity and inhibition of the beta-carbonic anhydrases (CAs, EC 4.2.1.1) from the pathogenic fungi Candida albicans (Nce103) and Cryptococcus neoformans (Can2) with inorganic anions such as halogenides, pseudohalogenides, bicarbonate, carbonate, nitrate, nitrite, hydrogen sulfide, bisulfite, perchlorate, sulfate were investigated. The two enzymes showed appreciable CO(2) hydrase activity (k(cat) in the range of (3.9-8.0)x10(5)s(-1), and k(cat)/K(m) in the range of (4.3-9.7)x10(7)M(-1)s(-1)). Can2 was weakly inhibited by cyanide and sulfamic acid (K(I)s of 8.22-13.56 mM), while all other anions displayed more potent inhibition. Nce103 was strongly inhibited by cyanide and carbonate (K(I)s of 10-11 microM), and weakly inhibited by sulfate, phenylboronic, and phenyl arsonic acid (K(I)s of 14.15-30.85 mM). These data demonstrate that pathogenic, fungal beta-CAs may be targets for the development of antifungals that have a novel mechanism of action.

  20. [State of Fungal Lipases of Rhizopus microsporus, Penicillium sp. and Oospora lactis in Border Layers Water-Solid Phase and Factors Affecting Catalytic Properties of Enzymes].

    Science.gov (United States)

    Khasanov, Kh T; Davranov, K; Rakhimov, M M

    2015-01-01

    We demonstrated that a change in the catalytic activity of fungal lipases synthesized by Rhizopus microsporus, Penicillium sp. and Oospora lactis and their ability to absorb on different sorbents depended on the nature of groups on the solid phase surface in the model systems water: lipid and water: solid phase. Thus, the stability of Penicillium sp. lipases increased 85% in the presence ofsorsilen or DEAE-cellulose, and 55% of their initial activity respectively was preserved. In the presence of silica gel and CM-cellulose, a decreased rate of lipid hydrolysis by Pseudomonas sp. enzymes was observed in water medium, and the hydrolysis rate increased by 2.4 and 1.5 times respectively in the presence of aminoaerosil and polykefamid. In an aqueous-alcohol medium, aminoaerosil and polykefamid decreased the rate of substrate hydrolysis by more than 30 times. The addition of aerosil to aqueous and aqueous-alcohol media resulted in an increase in the hydrolysis rate by 1.2-1.3 times. Sorsilen stabilized Penicillium sp. lipase activity at 40, 45, 50 and 55 degrees C. Either stabilization or inactivation of lipases was observed depending on the pH of the medium and the nature of chemical groups localized on the surface of solid phase. The synthetizing activity of lipases also changed depending on the conditions.

  1. Preparation and Characterization of Enzyme Compartments in UV-Cured Polyurethane-Based Materials and Their Application in Enzymatic Reactions

    Directory of Open Access Journals (Sweden)

    Diana Uhrich

    2017-11-01

    Full Text Available The preparation and characterization of UV-cured polyurethane-based materials for the mild inclusion immobilization of enzymes was investigated. Full curing of the polymer precursor/enzyme solution mixture was realized by a short irradiation with UV-light at ambient temperatures. The included aqueous enzyme solution remains highly dispersed in the polymer material with an even size distribution throughout the polymer material. The presented concept provides stable enzyme compartments which were applied for an alcohol dehydrogenase-catalyzed reduction reaction in organic solvents. Cofactor regeneration was achieved by a substrate-coupled approach via 2-propanol or an enzyme-coupled approach by a glucose dehydrogenase. This reaction concept can also be used for a simultaneous application of contrary biocatalytic reaction conditions within an enzymatic cascade reaction. Independent polymer-based reaction compartments were provided for two incompatible enzymatic reaction systems (alcohol dehydrogenase and hydroxynitrile lyase, while the relevant reactants diffuse between the applied compartments.

  2. Gelatin hydrolysate from blacktip shark skin prepared using papaya latex enzyme: Antioxidant activity and its potential in model systems.

    Science.gov (United States)

    Kittiphattanabawon, Phanat; Benjakul, Soottawat; Visessanguan, Wonnop; Shahidi, Fereidoon

    2012-12-01

    Antioxidant activities of gelatin hydrolysates from blacktip shark skin prepared using papaya latex enzyme with different degrees of hydrolysis (DHs: 10%, 20%, 30% and 40%) were evaluated. All antioxidant activity indices of hydrolysates increased with increasing DH (Pshark skin (40%DH) can potentially be used as an alternative source of natural antioxidants. Copyright © 2012 Elsevier Ltd. All rights reserved.

  3. A Biocatalytic One-Pot Approach for the Preparation of Lignin Oligomers Using an Oxidase/Peroxidase Cascade Enzyme System

    NARCIS (Netherlands)

    Habib, Mohamed H. M.; Deuss, Peter J.; Loncar, Nikola; Trajkovic, Milos; Fraaije, Marco W.

    2017-01-01

    Synthetic lignin was prepared biocatalytically in a one-pot, two-step reaction using an oxidase/peroxidase cascade enzyme system. Using eugenol in combination with eugenol oxidase and a peroxidase, lignin-like material was produced. The cascade reaction takes advantage of the ability of the oxidase

  4. Preparation of salted meat products, e.g. cured bacon - by injecting liquid comprising meat proteins hydrolysed with enzymes

    DEFF Research Database (Denmark)

    1997-01-01

    Preparation of salted meat products comprises the following:(1) meat is chopped into fine pieces and mixed with water to form a slurry; (2) enzymes hydrolyse proteins in the meat; (3) adding a culture to the resulting medium, which comprises short peptide chains or amino acids; (4) forming...... flavourings as the culture is growing, and (5) injecting the liquid into pieces of meat....

  5. Preparation of Coaxial-Electrospun Poly[bis(p-methylphenoxy]phosphazene Nanofiber Membrane for Enzyme Immobilization

    Directory of Open Access Journals (Sweden)

    Xiao-Jun Huang

    2012-11-01

    Full Text Available A core/sheath nanofiber membrane with poly[bis(p-methylphenoxy]phospha-zene (PMPPh as the sheath and easily spinnable polyacrylonitrile (PAN as the core was prepared via a coaxial electrospinning process. Field-emission scanning electron microscopy and transmission electron microscopy were used to characterize the morphology of the nanofiber membrane. It was found that the concentration of the PAN spinning solution and the ratio of the core/sheath solution flow rates played a decisive role in the coaxial electrospinning process. In addition, the stabilized core/sheath PMPPh nanofiber membrane was investigated as a support for enzyme immobilization because of its excellent biocompatibility, high surface/volume ratio, and large porosity. Lipase from Candida rugosa was immobilized on the nanofiber membrane by adsorption. The properties of the immobilized lipase on the polyphosphazene nanofiber membrane were studied and compared with those of a PAN nanofiber membrane. The results showed that the adsorption capacity (20.4 ± 2.7 mg/g and activity retention (63.7% of the immobilized lipase on the polyphosphazene nanofiber membrane were higher than those on the PAN membrane.

  6. Studies on the preparation of immobilized enzymes by radio-polymerization, 10

    International Nuclear Information System (INIS)

    Amarakone, S.P.; Hayashi, Toru; Kawashima, Koji.

    1983-01-01

    β-Galactosidase of E. coli origin was immobilized in the form of beads by the radiopolymerization of different combinations of monomers using a gamma irradiation technique. With the dialysed enzyme, recoveries of over 300 % could be obtained on suitable monomer combinations containing magnesium and sodium acrylates. The recovery of the enzyme also depended on the irradiation time. The immobilized enzyme had better pH and temperature stability and was less affected by the presence of metal ions in the medium, compared to the native enzyme. The optimum pH and temperatures of the immobilized enzyme were different from those of the native enzyme and were 7.0 to 7.5 and 50 deg C respectively. The immobilized enzyme was used in a column for the continuous determination of lactose with a standard type autoanalyser. Good linearity could be observed even up to 3 % lactose in the sample. (author)

  7. Comprehensive utilization of activated sludge for the preparation of hydrolytic enzymes, polyhydroxyalkanoates, and water-retaining organic fertilizer.

    Science.gov (United States)

    Ni, He; Fan, Xiao-Min; Guo, Hao-Ning; Liang, Jian-Hua; Li, Qing-Rong; Yang, Liu; Li, Hui; Li, Hai-Hang

    2017-07-03

    The urban wastewater treatment industry produces a large amount of excess activated sludge which is mainly composed of microbial biomass and costly to be disposed. In this research, a comprehensive utilization of activated sludge was developed by sequentially extracting hydrolytic enzymes and polyhydroxyalkanoates (PHAs), and the residue was used to prepare water-retaining organic fertilizer. The sludge was extracted with fourfold H 2 O-containing 1% Triton X-100 with the yield of 66.7% protease activity. The enzyme solution was precipitated in 80% acetone and vacuum dried at 40°C at the dried enzyme yield of 2.4 g/kg wet sludge. The enzyme product contains collagenase, lipase, amylase, and cellulase activities, which are good compound enzymes to feed. The PHAs were extracted with 30% sodium hypoclorite:chloroform (1:3). The PHA solution was decolored and dried, and pure white PHAs were obtained at the yield of 70.1 g/kg wet sludge. The residue was used to prepare water-retaining organic fertilizer at the optimal condition. The fertilizer absorbs 131.3-fold distilled water and had good performance in water retention and can effectively slow down the loss of soil moisture when added into soil. This work provides a simple and practical approach for comprehensive utilizing activated sludge with significant economic benefits.

  8. Enzymatic formation of compound-K from ginsenoside Rb1 by enzyme preparation from cultured mycelia of Armillaria mellea.

    Science.gov (United States)

    Upadhyaya, Jitendra; Kim, Min-Ji; Kim, Young-Hoi; Ko, Sung-Ryong; Park, Hee-Won; Kim, Myung-Kon

    2016-04-01

    Minor saponins or human intestinal bacterial metabolites, such as ginsenosides Rg3, F2, Rh2, and compound K, are more pharmacologically active than major saponins, such as ginsenosides Rb1, Rb2, and Rc. In this work, enzymatic hydrolysis of ginsenoside Rb1 was studied using enzyme preparations from cultured mycelia of mushrooms. Mycelia of Armillaria mellea, Ganoderma lucidum, Phellinus linteus, Elfvingia applanata, and Pleurotus ostreatus were cultivated in liquid media at 25°C for 2 wk. Enzyme preparations from cultured mycelia of five mushrooms were obtained by mycelia separation from cultured broth, enzyme extraction, ammonium sulfate (30-80%) precipitation, dialysis, and freeze drying, respectively. The enzyme preparations were used for enzymatic hydrolysis of ginsenoside Rb1. Among the mushrooms used in this study, the enzyme preparation from cultured mycelia of A. mellea (AMMEP) was found to convert ginsenoside Rb1 into compound K with a high yield, while those from G. lucidum, P. linteus, E. applanata, and P. ostreatus produced remarkable amounts of ginsenoside Rd from ginsenoside Rb1. The enzymatic hydrolysis pathway of ginsenoside Rb1 by AMMEP was Rb1 → Rd → F2 → compound K. The optimum reaction conditions for compound K formation from ginsenoside Rb1 were as follows: reaction time 72-96 h, pH 4.0-4.5, and temperature 45-55°C. AMMEP can be used to produce the human intestinal bacterial metabolite, compound K, from ginsenoside Rb1 with a high yield and without food safety issues.

  9. Preparative studies on the isolation of an enzyme associated with carthamin synthesis in Carthamus tinetorius L.

    Directory of Open Access Journals (Sweden)

    Takao Homma

    2014-01-01

    Full Text Available Preliminary operations for the isolation of an enzyme associated with cathamin synthesis (carthamin-synthesizing enzyme were done with a soluble extract of safflower seedlings. Ethanol and acetone seriously affected the solubility and the activity of the enzyme. Ammonium sulfate fractionation, calcium acetate precipitation and protamine sulfate treatment were all useful techniques for removing inert protein from the crude extract however, the activity of the carthamin-synthesizing enzyme could not be separated from that of native polyphenol oxidase or peroxidase. Partial purification of the carthamin-synthesizing enzyme could be achieved by filtration of a crude extract through Sephadex G-100 gel after treating it with ammonium sulfate, calcium acetate and protamine sulfate. -the purified enzyme reacted possitively with precarthamin at pH 4.8 in 50.0 mM acetate buffer. The synthesized product was identified as carthamin by checking its Rf values on silica gel plates developed with three different developing solvents and UV absorption spectral pattern in methanol. The enzyme sample was unstable on storage and stabilization of the sample was tested by applying many procedures. Carthamin-synthesizing enzyme had low affinity to celite 535. No carthamin was found in an incubation medium containing precarthamin and an authentic sample of polyphenol oxidase or peroxidase. Carthamin could not be converted to any blackish catabolites through polyphenol oxidase- or peroxidase-catalyzed oxidative reaction. Changes in the activities of three different enzymes, carthamin-synthesizing enzyme, polyphenol oxidase and peroxidase, were investigated in the extract from both etiolated and green seedlings of safflower and the data were discussed in relation to the catalytic properties of the enzymes.

  10. Preparation and evaluation of a coumarin library towards the inhibitory activity of the enzyme gGAPDH from Trypanosoma cruzi

    International Nuclear Information System (INIS)

    Alvim Junior, Joel; Dias, Ricardo L.A.; Correa, Arlene G.; Castilho, Marcelo S.; Oliva, Glaucius

    2005-01-01

    Chagas' disease, caused by Trypanosoma cruzi, is endemic in 15 countries in Latin America. In this work a library of 38 coumarins was prepared in solution phase and evaluated against T. cruzi glycolytic enzyme glyceraldehyde-3-phosphate-dehydrogenase (gGAPDH). The synthetic route was based on the Knoevenagel condensation of different 2-hydroxybenzaldehydes with Meldrum's acid or diethyl malonate, followed by O-alkylation and/or transesterification reactions. Among the prepared coumarins, the best values obtained to inhibit 50% of the enzymatic activity range from 80 to 130 μM. (author)

  11. Preparation and evaluation of a coumarin library towards the inhibitory activity of the enzyme gGAPDH from Trypanosoma cruzi

    Energy Technology Data Exchange (ETDEWEB)

    Alvim Junior, Joel; Dias, Ricardo L.A.; Correa, Arlene G. [Universidade Federal de Sao Carlos, SP (Brazil). Dept. de Quimica]. E-mail: agcorrea@power.ufscar.br; Castilho, Marcelo S.; Oliva, Glaucius [Sao Paulo Univ., Sao Carlos, SP (Brazil). Inst. de Fisica

    2005-07-15

    Chagas' disease, caused by Trypanosoma cruzi, is endemic in 15 countries in Latin America. In this work a library of 38 coumarins was prepared in solution phase and evaluated against T. cruzi glycolytic enzyme glyceraldehyde-3-phosphate-dehydrogenase (gGAPDH). The synthetic route was based on the Knoevenagel condensation of different 2-hydroxybenzaldehydes with Meldrum's acid or diethyl malonate, followed by O-alkylation and/or transesterification reactions. Among the prepared coumarins, the best values obtained to inhibit 50% of the enzymatic activity range from 80 to 130 {mu}M. (author)

  12. Rapid delivery of diazepam from supersaturated solutions prepared using prodrug/enzyme mixtures: toward intranasal treatment of seizure emergencies.

    Science.gov (United States)

    Kapoor, Mamta; Winter, Tate; Lis, Lev; Georg, Gunda I; Siegel, Ronald A

    2014-05-01

    Current treatments for seizure emergencies, such as status epilepticus, include intravenous or rectal administration of benzodiazepines. While intranasal delivery of these drugs is desirable, the small volume of the nasal cavity and low drug solubility pose significant difficulties. Here, we prepared supersaturated diazepam solutions under physiological conditions and without precipitation, using a prodrug/enzyme system. Avizafone, a peptide prodrug of diazepam, was delivered with--Aspergillus oryzae (A.O.) protease, an enzyme identified from a pool of hydrolytic enzymes in assay buffer, pH 7.4 at 32°C. This enzyme converted avizafone to diazepam at supersaturated concentrations. In vitro permeability studies were performed at various prodrug/enzyme ratios using Madin-Darby canine kidney II-wild type (MDCKII-wt) monolayers, a representative model of the nasal epithelium. Monolayer integrity was examined using TEER measurement and the lucifer yellow permeability assay. Prodrug/drug concentrations were measured using HPLC. Enzyme kinetics with avizafone-protease mixtures revealed K(M) = 1,501 ± 232 μM and V(max) = 1,369 ± 94 μM/s. Prodrug-protease mixtures, when co-delivered apically onto MDCKII-wt monolayers, showed 2-17.6-fold greater diazepam flux (S = 1.3-15.3) compared to near-saturated diazepam (S = 0.7). Data for prodrug conversion upstream (apical side) and drug permeability downstream (basolateral side) fitted reasonably well to a previously developed in vitro two compartment pharmacokinetic model. Avizafone-protease mixtures resulted in supersaturated diazepam in less than 5 min, with the rate and extent of supersaturation determined by the prodrug/enzyme ratio. Together, these results suggest that an intranasal avizafone-protease system may provide a rapid and alternative means of diazepam delivery.

  13. Protoplast preparation from monokaryotic mycelium of Pleurotus sajor-caju using lysing enzyme

    International Nuclear Information System (INIS)

    Hassan Hamdani Mutaat; Mat Rasol Awang

    2004-01-01

    The objective of this study was to determine the optimum parameters of the factors influencing protoplast isolation from monokaryotic mycelium of Pleurotus sajor-caju using lysing enzyme from Trichoderma harzianurm. The study was conducted by manipulating the variables of the factors affecting protoplast isolation, such as age of mycelium culture, period for lysing of mycelium, concentration of lysing enzyme and concentration of osmotic stabilizer. The highest protoplast yield of 8.3 x 104 protoplast/ml was achieved when a 3-day P. sajor-caju mycelium, cultured statically, was incubated for 3 hours in a lytic mixture containing 7.5 mg/ml lysing enzyme and 1.2 M ammonium sulfate as osmotic stabilizer. This protoplast yield, however, is insufficient for regeneration and protoplast fusion works. (Author)

  14. Low-cost, easy-to-prepare magnetic chitosan microparticles for enzymes immobilization

    Czech Academy of Sciences Publication Activity Database

    Pospišková, K.; Šafařík, Ivo

    2013-01-01

    Roč. 96, č. 2 (2013), s. 545-548 ISSN 0144-8617 R&D Projects: GA MŠk(CZ) LD13021 Institutional support: RVO:67179843 Keywords : Chitosan * Magnetite * Microwave irradiation * Enzymes immobilization Subject RIV: CE - Biochemistry Impact factor: 3.916, year: 2013

  15. Changes in activity of industrial enzyme preparations irradiated with sterilizing doses. Part of a coordinated programme on factors influencing the utilization of food irradiation process

    International Nuclear Information System (INIS)

    Bachman, S.

    1984-03-01

    Experiments were carried out to investigate the efficacy of irradiation to sterilize enzyme preparations. Irradiation doses up to 25 kGy caused no changes in basic organoleptic properties of commercial rennin preparations. Dose rate (from 0.5 to 13.5 kGy/hr) has no influence on the changes in enzyme activity during the storage period of 3 months. Doses ranging from 8 to 12 kGy are sufficient to sterilize commercial enzyme preparations. Non-purified, crude rennin preparations appear to be more resistant to radiation than purified samples. Rennin preparations purified by dialysis and treated with 25 kGy resulted in a reduction of activity of 20%. The activity of preparations purified by gel filtration was reduced to 50% when treated with the same dose

  16. Preparation, crystallization and preliminary crystallographic analysis of old yellow enzyme from Trypanosoma cruzi

    Energy Technology Data Exchange (ETDEWEB)

    Sugiyama, Shigeru [Maruwa Foods Co. Ltd, Tsutsui-cho 170-1, Yamatokoriyama, Nara 639-1123 (Japan); Tokuoka, Keiji [Department of Applied Chemistry, Graduate School of Engineering, Osaka University, 2-1 Yamada-Oka, Suita, Osaka 565-0871 (Japan); Uchiyama, Nahoko [Department of Molecular Behavioral Biology, Osaka Bioscience Institute, Osaka 565-0874 (Japan); Okamoto, Naoki; Okano, Yousuke; Matsumura, Hiroyoshi [Department of Applied Chemistry, Graduate School of Engineering, Osaka University, 2-1 Yamada-Oka, Suita, Osaka 565-0871 (Japan); Inaka, Koji [Maruwa Foods Co. Ltd, Tsutsui-cho 170-1, Yamatokoriyama, Nara 639-1123 (Japan); Urade, Yoshihiro [Department of Molecular Behavioral Biology, Osaka Bioscience Institute, Osaka 565-0874 (Japan); Inoue, Tsuyoshi, E-mail: inouet@chem.eng.osaka-u.ac.jp [Department of Applied Chemistry, Graduate School of Engineering, Osaka University, 2-1 Yamada-Oka, Suita, Osaka 565-0871 (Japan); Maruwa Foods Co. Ltd, Tsutsui-cho 170-1, Yamatokoriyama, Nara 639-1123 (Japan)

    2007-10-01

    Old yellow enzyme from Trypanosoma cruzi, has been crystallized using the hanging-drop vapour-diffusion method. Old yellow enzyme (OYE) is an NADPH oxidoreductase that contains a flavin mononucleotide as a prosthetic group. The OYE from Trypanosoma cruzi, which produces prostaglandin F{sub 2α}, a potent mediator of various physiological and pathological processes, from prostaglandin H2. The protein was recombinantly expressed and purified from Escherichia coli and was crystallized using the hanging-drop vapour-diffusion method. The crystal belongs to the monoclinic space group P2{sub 1}, with unit-cell parameters a = 56.3, b = 78.8, c = 78.8 Å, β = 93.4° and two molecules per asymmetric unit. The crystals were suitable for X-ray crystallographic studies and diffracted to 1.70 Å resolution. A Patterson search method is in progress using the structure of OYE from Pseudomonas putida as a starting model.

  17. Utilization of newly developed immobilized enzyme reactors for preparation and study of immunoglobulin G fragments

    Czech Academy of Sciences Publication Activity Database

    Korecká, L.; Bílková, Z.; Holčapek, M.; Královský, J.; Beneš, Milan J.; Lenfeld, Jiří; Minc, N.; Cecal, R.; Viovy, J.-L.; Przybylski, M.

    2004-01-01

    Roč. 808, č. 1 (2004), s. 15-24 ISSN 1570-0232. [International Symposium on Polymer Design for BioSeparation and Nanobiotechnology /8./. Compiegne, 27.11.2003-29.11.2003] Grant - others:GA ČR(CZ) GA203/02/0023 Program:GA Institutional research plan: CEZ:AV0Z4050913 Keywords : immobilized enzyme reactors * immunoglobulin G Subject RIV: CE - Biochemistry Impact factor: 2.176, year: 2004

  18. Fungal Diseases

    Science.gov (United States)

    ... to other illnesses such as the flu or tuberculosis. Some fungal diseases like fungal meningitis and bloodstream ... prevención Fuentes Diagnóstico y pruebas Tratamiento Profesionales de la salud Estadísticas Blastomycosis Definition Symptoms Risk & Prevention Sources ...

  19. Fungal Infections

    Science.gov (United States)

    ... Body Mind Sexual Health Food & Fitness Diseases & Conditions Infections Drugs & Alcohol School & Jobs Sports Expert Answers (Q&A) Staying Safe Videos for Educators Search English Español Fungal Infections KidsHealth / For Kids / Fungal Infections What's in this ...

  20. Changes in Land Use System and Environmental Factors Affect Arbuscular Mycorrhizal Fungal Density and Diversity, and Enzyme Activities in Rhizospheric Soils of Acacia senegal (L.) Willd.

    OpenAIRE

    Ndoye, Fatou; Kane, Aboubacry; Ngonkeu Mangaptché, Eddy Léonard; Bakhoum, Niokhor; Sanon, Arsène; Diouf, Diégane; Sy, Mame Ourèye; Baudoin, Ezékiel; Noba, Kandioura; Prin, Yves

    2012-01-01

    The responses of the soil microbial community features associated to the legume tree Acacia senegal (L.) Willd. including both arbuscular mycorrhizal fungal (AMF) diversity and soil bacterial functions, were investigated under contrasting environmental conditions. Soil samples were collected during dry and rainy seasons in two contrasting rainfall sites of Senegal (Dahra and Goudiry, in arid and semiarid zone, resp.). Soils were taken from the rhizosphere of A. senegal both in plantation and ...

  1. The Effects of Opium Addiction on the Immune System Function in Patients with Fungal Infection.

    Science.gov (United States)

    Ayatollahi-Mousavi, Seyyed Amin; Asadikaram, Gholamreza; Nakhaee, Nouzar; Izadi, Alireza; Keikha, Nasser

    2016-01-01

    The use of narcotics such as opium exposes addicts as susceptible targets of different diseases so that they might easily be exposed to different diseases such as fungal infections. The present study aimed to investigate the effects of addiction to opium and fungal infection on plasma levels of certain cytokines including interleukin-4 (IL-4), IL-6, IL-17, Interferon gamma (IFN-γ) and transforming growth factor-β (TGF-β). Present study included 72 individuals who were divided into 4 groups: 1) opium-addicted with fungal infection; 2) opium-addicted without fungal infection; 3) non-opium-addicted with fungal infection; and 4) normal individuals (non-opium-addicted and non-fungal infection). The fungal samples, after being detected and confirmed by a physician, were prepared based on clinical symptoms and then analyzed by direct smear and culture method. The measurement of the plasma level of cytokines was done by enzyme-linked immunosorbent assay (ELISA) method. The comparison of the mean of the plasma level of cytokines showed that addiction to opium and fungal infection had significant effect on the plasma levels of IL-17, IFN-γ, TGF-β cytokines in all studied groups. The interaction of addiction to opium and fungal infection was only significant in the case of plasma level of IL-6. Addiction to opium and fungal infection, either separately or simultaneously, poses significant effect on the immune system and causes disorders in the cytokine network and the immune system and also provides a suitable environment for fungal infection.

  2. Assessment of the Effects of Intermittent Mixing On Solid-State Fermentation for Biomass-Degrading Enzymes Production by Different Fungal Strains

    OpenAIRE

    Cristiane S. Farinas

    2015-01-01

    The use of solid-state fermentation (SSF) for the production of industrial enzymes has received increasing attention over the years. However, the implementation of large-scale SSF processes requires an understanding of the effects of mixing on microorganism growth and product formation. This paper describes a systematic comparison of the effects of intermittent mixing on SSF, in terms of the production of biomass-degrading enzymes (endoglucanase, β-glucosidase, and xylanase) by different fung...

  3. Preparation of antioxidant enzymatic hydrolysates from honeybee-collected pollen using plant enzymes.

    Science.gov (United States)

    Marinova, Margarita D; Tchorbanov, Bozhidar P

    2011-01-09

    Enzymatic hydrolysates of honeybee-collected pollen were prepared using food-grade proteinase and aminopeptidases entirely of plant origin. Bromelain from pineapple stem was applied (8 mAU/g substrate) in the first hydrolysis stage. Aminopeptidase (0.05 U/g substrate) and proline iminopeptidase (0.03 U/g substrate) from cabbage leaves (Brassica oleracea var. capitata), and aminopeptidase (0.2 U/g substrate) from chick-pea cotyledons (Cicer arietinum L.) were involved in the additional hydrolysis of the peptide mixtures. The degree of hydrolysis (DH), total phenolic contents, and protein contents of these hydrolysates were as follows: DH (about 20-28%), total phenolics (15.3-27.2 μg/mg sample powder), and proteins (162.7-242.8 μg/mg sample powder), respectively. The hydrolysates possessed high antiradical scavenging activity determined with DPPH (42-46% inhibition). The prepared hydrolysates of bee-collected flower pollen may be regarded as effective natural and functional dietary food supplements due to their remarkable content of polyphenol substances and significant radical-scavenging capacity with special regard to their nutritional-physiological implications.

  4. Preparation of Antioxidant Enzymatic Hydrolysates from Honeybee-Collected Pollen Using Plant Enzymes

    Directory of Open Access Journals (Sweden)

    Margarita D. Marinova

    2010-01-01

    Full Text Available Enzymatic hydrolysates of honeybee-collected pollen were prepared using food-grade proteinase and aminopeptidases entirely of plant origin. Bromelain from pineapple stem was applied (8 mAU/g substrate in the first hydrolysis stage. Aminopeptidase (0.05 U/g substrate and proline iminopeptidase (0.03 U/g substrate from cabbage leaves (Brassica oleracea var. capitata, and aminopeptidase (0.2 U/g substrate from chick-pea cotyledons (Cicer arietinum L. were involved in the additional hydrolysis of the peptide mixtures. The degree of hydrolysis (DH, total phenolic contents, and protein contents of these hydrolysates were as follows: DH (about 20–28%, total phenolics (15.3–27.2 μg/mg sample powder, and proteins (162.7–242.8 μg/mg sample powder, respectively. The hydrolysates possessed high antiradical scavenging activity determined with DPPH (42–46% inhibition. The prepared hydrolysates of bee-collected flower pollen may be regarded as effective natural and functional dietary food supplements due to their remarkable content of polyphenol substances and significant radical-scavenging capacity with special regard to their nutritional-physiological implications.

  5. Biotechnological approaches to develop bacterial chitinases as a bioshield against fungal diseases of plants.

    Science.gov (United States)

    Neeraja, Chilukoti; Anil, Kondreddy; Purushotham, Pallinti; Suma, Katta; Sarma, Pvsrn; Moerschbacher, Bruno M; Podile, Appa Rao

    2010-09-01

    Fungal diseases of plants continue to contribute to heavy crop losses in spite of the best control efforts of plant pathologists. Breeding for disease-resistant varieties and the application of synthetic chemical fungicides are the most widely accepted approaches in plant disease management. An alternative approach to avoid the undesired effects of chemical control could be biological control using antifungal bacteria that exhibit a direct action against fungal pathogens. Several biocontrol agents, with specific fungal targets, have been registered and released in the commercial market with different fungal pathogens as targets. However, these have not yet achieved their full commercial potential due to the inherent limitations in the use of living organisms, such as relatively short shelf life of the products and inconsistent performance in the field. Different mechanisms of action have been identified in microbial biocontrol of fungal plant diseases including competition for space or nutrients, production of antifungal metabolites, and secretion of hydrolytic enzymes such as chitinases and glucanases. This review focuses on the bacterial chitinases that hydrolyze the chitinous fungal cell wall, which is the most important targeted structural component of fungal pathogens. The application of the hydrolytic enzyme preparations, devoid of live bacteria, could be more efficacious in fungal control strategies. This approach, however, is still in its infancy, due to prohibitive production costs. Here, we critically examine available sources of bacterial chitinases and the approaches to improve enzymatic properties using biotechnological tools. We project that the combination of microbial and recombinant DNA technologies will yield more effective environment-friendly products of bacterial chitinases to control fungal diseases of crops.

  6. Overproduction of ligninolytic enzymes

    Science.gov (United States)

    Elisashvili, Vladimir; Kachlishvili, Eva; Torok, Tamas

    2014-06-17

    Methods, compositions, and systems for overproducing ligninolytic enzymes from the basidiomycetous fungus are described herein. As described, the method can include incubating a fungal strain of Cerrena unicolor IBB 303 in a fermentation system having growth medium which includes lignocellulosic material and then cultivating the fungal strain in the fermentation system under conditions wherein the fungus expresses the ligninolytic enzymes. In some cases, the lignocellulosic material is mandarin peel, ethanol production residue, walnut pericarp, wheat bran, wheat straw, or banana peel.

  7. Performance of hemicellulolytic enzymes in culture supernatants from a wide range of fungi on insoluble wheat straw and corn fiber fractions.

    Science.gov (United States)

    van Gool, M P; Toth, K; Schols, H A; Szakacs, G; Gruppen, H

    2012-06-01

    Filamentous fungi are a good source of hemicellulolytic enzymes for biomass degradation. Enzyme preparations were obtained as culture supernatants from 78 fungal isolates grown on wheat straw as carbon source. These enzyme preparations were utilized in the hydrolysis of insoluble wheat straw and corn fiber xylan rich fractions. Up to 14% of the carbohydrates in wheat straw and 34% of those in corn fiber were hydrolyzed. The degree of hydrolysis by the enzymes depended on the origin of the fungal isolate and on the complexity of the substrate to be degraded. Penicillium, Trichoderma or Aspergillus species, and some non-identified fungi proved to be the best producers of hemicellulolytic enzymes for degradation of xylan rich materials. This study proves that the choice for an enzyme preparation to efficiently degrade a natural xylan rich substrate, is dependent on the xylan characteristics and could not be estimated by using model substrates. Copyright © 2012 Elsevier Ltd. All rights reserved.

  8. The influence of Saccharomyces cerevisiae enzyme ratio on preparation virgin coconut oil for candidate in-house reference materials

    Science.gov (United States)

    Rohyami, Yuli; Anjani, Rafika Debby; Purwanti, Napthalina Putri

    2017-03-01

    Virgin coconut oil is an excellent product which has result of oil processing business opportunities in the international market. Standardization of virgin coconut oil necessary to satisfy the requirements industry needs. This research is expected as procedure preparation of reference materials. Preparation of virgin coconut oil by Sacharomycescerevisiaeenzyme. Based on the results of this study concluded that the ratio of Saccharomyces cerevisiae can affect the yield of virgin coconut oil produced. The preparation of virgin coconut oil enzymatically using a variety of mass ratio of 0.001 to 0.006% is obtained yield average of 12.40%. The optimum separation of virgin coconut oil on the use of enzymes with a mass ratio of 0.002%. The average water content at a ratio of 0.002% is 0.04 % with a value of uncertainty is 0.005%. The average iodine number in virgin coconut oil produced is 2.4403 ± 0,1974 grams of iodine per 100 grams of oil and optimum iodine number is obtained from the manufacturing process virgin coconut oil with a ratio of 0.006% Saccharomyces cerevisiae. Sacharomycescerevisiae with a ratio of 0.002% results virgin coconut oil with acid number 0.3068 ± 0.1098%. The peroxide value of virgin coconut oil between 0.0108 ± 0.009 to 0.0114 ± 0015milli-equivalent per kilograms. Organoleptic test results and test chemical parameters can be used as the test data that can be developed in prototype preparation of candidate in-house reference material in the testing standards of quality virgin coconut oil.

  9. Preparation of reusable bioreactors using reversible immobilization of enzyme on monolithic porous polymer support with attached gold nanoparticles.

    Science.gov (United States)

    Lv, Yongqin; Lin, Zhixing; Tan, Tianwei; Svec, Frantisek

    2014-01-01

    Porcine lipase has been reversibly immobilized on a monolithic polymer support containing thiol functionalities prepared within confines of a fused silica capillary and functionalized with gold nanoparticles. Use of gold nanoparticles enabled rejuvenation of the activity of the deactivated reactor simply by stripping the inactive enzyme from the nanoparticles using 2-mercaptoethanol and subsequent immobilization of fresh lipase. This flow through enzymatic reactor was then used to catalyze the hydrolysis of glyceryl tributyrate (tributyrin). The highest activity was found within a temperature range of 37-40°C. The reaction kinetics is characterized by Michaelis-Menten constant, Km  = 10.9 mmol/L, and maximum reaction rate, Vmax  = 5.0 mmol/L min. The maximum reaction rate for the immobilized enzyme is 1,000 times faster compared to lipase in solution. The fast reaction rate enabled to achieve 86.7% conversion of tributyrin in mere 2.5 min and an almost complete conversion in 10 min. The reactor lost only less than 10% of its activity even after continuous pumping through it a solution of substrate equaling 1,760 reactor volumes. Finally, potential application of this enzymatic reactor was demonstrated with the transesterification of triacylglycerides from kitchen oil to fatty acid methyl esters thus demonstrating the ability of the reactor to produce biodiesel. © 2013 Wiley Periodicals, Inc.

  10. Early-branching Gut Fungi Possess A Large, And Comprehensive Array Of Biomass-Degrading Enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Solomon, Kevin V.; Haitjema, Charles; Henske, John K.; Gilmore, Sean P.; Borges-Rivera, Diego; Lipzen, Anna; Brewer, Heather M.; Purvine, Samuel O.; Wright, Aaron T.; Theodorou, Michael K.; Grigoriev, Igor V.; Regev, Aviv; Thompson, Dawn; O' Malley, Michelle A.

    2016-03-11

    The fungal kingdom is the source of almost all industrial enzymes in use for lignocellulose bioprocessing. Its more primitive members, however, remain relatively unexploited. We developed a systems-level approach that integrates RNA-Seq, proteomics, phenotype and biochemical studies of relatively unexplored early-branching free-living fungi. Anaerobic gut fungi isolated from herbivores produce a large array of biomass-degrading enzymes that synergistically degrade crude, unpretreated plant biomass, and are competitive with optimized commercial preparations from Aspergillus and Trichoderma. Compared to these model platforms, gut fungal enzymes are unbiased in substrate preference due to a wealth of xylan-degrading enzymes. These enzymes are universally catabolite repressed, and are further regulated by a rich landscape of noncoding regulatory RNAs. Furthermore, we identified several promising sequence divergent enzyme candidates for lignocellulosic bioprocessing.

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

    Science.gov (United States)

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

    2016-05-01

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

  12. The fungal ?-aminoadipate pathway for lysine biosynthesis requires two enzymes of the aconitase family for the isomerization of homocitrate to homoisocitrate

    OpenAIRE

    Fazius, Felicitas; Shelest, Ekaterina; Gebhardt, Peter; Brock, Matthias

    2012-01-01

    Fungi produce ?-aminoadipate, a precursor for penicillin and lysine via the ?-aminoadipate pathway. Despite the biotechnological importance of this pathway, the essential isomerization of homocitrate via homoaconitate to homoisocitrate has hardly been studied. Therefore, we analysed the role of homoaconitases and aconitases in this isomerization. Although we confirmed an essential contribution of homoaconitases from Saccharomyces cerevisiae and Aspergillus fumigatus, these enzymes only cataly...

  13. Production by Tobacco Transplastomic Plants of Recombinant Fungal and Bacterial Cell-Wall Degrading Enzymes to Be Used for Cellulosic Biomass Saccharification

    Directory of Open Access Journals (Sweden)

    Paolo Longoni

    2015-01-01

    Full Text Available Biofuels from renewable plant biomass are gaining momentum due to climate change related to atmospheric CO2 increase. However, the production cost of enzymes required for cellulosic biomass saccharification is a major limiting step in this process. Low-cost production of large amounts of recombinant enzymes by transgenic plants was proposed as an alternative to the conventional microbial based fermentation. A number of studies have shown that chloroplast-based gene expression offers several advantages over nuclear transformation due to efficient transcription and translation systems and high copy number of the transgene. In this study, we expressed in tobacco chloroplasts microbial genes encoding five cellulases and a polygalacturonase. Leaf extracts containing the recombinant enzymes showed the ability to degrade various cell-wall components under different conditions, singly and in combinations. In addition, our group also tested a previously described thermostable xylanase in combination with a cellulase and a polygalacturonase to study the cumulative effect on the depolymerization of a complex plant substrate. Our results demonstrate the feasibility of using transplastomic tobacco leaf extracts to convert cell-wall polysaccharides into reducing sugars, fulfilling a major prerequisite of large scale availability of a variety of cell-wall degrading enzymes for biofuel industry.

  14. Production by Tobacco Transplastomic Plants of Recombinant Fungal and Bacterial Cell-Wall Degrading Enzymes to Be Used for Cellulosic Biomass Saccharification.

    Science.gov (United States)

    Longoni, Paolo; Leelavathi, Sadhu; Doria, Enrico; Reddy, Vanga Siva; Cella, Rino

    2015-01-01

    Biofuels from renewable plant biomass are gaining momentum due to climate change related to atmospheric CO2 increase. However, the production cost of enzymes required for cellulosic biomass saccharification is a major limiting step in this process. Low-cost production of large amounts of recombinant enzymes by transgenic plants was proposed as an alternative to the conventional microbial based fermentation. A number of studies have shown that chloroplast-based gene expression offers several advantages over nuclear transformation due to efficient transcription and translation systems and high copy number of the transgene. In this study, we expressed in tobacco chloroplasts microbial genes encoding five cellulases and a polygalacturonase. Leaf extracts containing the recombinant enzymes showed the ability to degrade various cell-wall components under different conditions, singly and in combinations. In addition, our group also tested a previously described thermostable xylanase in combination with a cellulase and a polygalacturonase to study the cumulative effect on the depolymerization of a complex plant substrate. Our results demonstrate the feasibility of using transplastomic tobacco leaf extracts to convert cell-wall polysaccharides into reducing sugars, fulfilling a major prerequisite of large scale availability of a variety of cell-wall degrading enzymes for biofuel industry.

  15. Fungal keratitis

    Directory of Open Access Journals (Sweden)

    Sonal S Tuli

    2011-02-01

    Full Text Available Sonal S TuliUniversity of Florida, Gainesville, FL, USA  Clinical question: What is the most appropriate management of fungal keratitis?Results: Traditionally, topical Natamycin is the most commonly used medication for filamentous fungi while Amphotericin B is most commonly used for yeast. Voriconazole is rapidly becoming the drug of choice for all fungal keratitis because of its wide spectrum of coverage and increased penetration into the cornea.Implementation: Repeated debridement of the ulcer is recommended for the penetration of topical medications. While small, peripheral ulcers may be treated in the community, larger or central ulcers, especially if associated with signs suggestive of anterior chamber penetration should be referred to a tertiary center. Prolonged therapy for approximately four weeks is usually necessary.Keywords: fungal keratitis, keratomycosis, antifungal medications, debridement

  16. A novel class of fungal lipoxygenases

    NARCIS (Netherlands)

    Heshof, R.; Jylhä, S.; Haarmann, T.; Jørgensen, A.L.W.; Dalsgaard, T.K.; Graaff, de L.H.

    2014-01-01

    Lipoxygenases (LOXs) are well-studied enzymes in plants and mammals. However, fungal LOXs are less studied. In this study, we have compared fungal LOX protein sequences to all known characterized LOXs. For this, a script was written using Shell commands to extract sequences from the NCBI database

  17. Directed evolution of fungal laccases.

    Science.gov (United States)

    Maté, Diana; García-Ruiz, Eva; Camarero, Susana; Alcalde, Miguel

    2011-04-01

    Fungal laccases are generalists biocatalysts with potential applications that range from bioremediation to novel green processes. Fuelled by molecular oxygen, these enzymes can act on dozens of molecules of different chemical nature, and with the help of redox mediators, their spectrum of oxidizable substrates is further pushed towards xenobiotic compounds (pesticides, industrial dyes, PAHs), biopolymers (lignin, starch, cellulose) and other complex molecules. In recent years, extraordinary efforts have been made to engineer fungal laccases by directed evolution and semi-rational approaches to improve their functional expression or stability. All these studies have taken advantage of Saccharomyces cerevisiae as a heterologous host, not only to secrete the enzyme but also, to emulate the introduction of genetic diversity through in vivo DNA recombination. Here, we discuss all these endeavours to convert fungal laccases into valuable biomolecular platforms on which new functions can be tailored by directed evolution.

  18. Lignin-degrading enzyme activities.

    Science.gov (United States)

    Chen, Yi-ru; Sarkanen, Simo; Wang, Yun-Yan

    2012-01-01

    Over the past three decades, the activities of four kinds of enzyme have been purported to furnish the mechanistic foundations for macromolecular lignin depolymerization in decaying plant cell walls. The pertinent fungal enzymes comprise lignin peroxidase (with a relatively high redox potential), manganese peroxidase, an alkyl aryl etherase, and laccase. The peroxidases and laccase, but not the etherase, are expressed extracellularly by white-rot fungi. A number of these microorganisms exhibit a marked preference toward lignin in their degradation of lignocellulose. Interestingly, some white-rot fungi secrete both kinds of peroxidase but no laccase, while others that are equally effective express extracellular laccase activity but no peroxidases. Actually, none of these enzymes has been reported to possess significant depolymerase activity toward macromolecular lignin substrates that are derived with little chemical modification from the native biopolymer. Here, the assays commonly employed for monitoring the traditional fungal peroxidases, alkyl aryl etherase, and laccase are described in their respective contexts. A soluble native polymeric substrate that can be isolated directly from a conventional milled-wood lignin preparation is characterized in relation to its utility in next-generation lignin-depolymerase assays.

  19. Development of technology for gourmet smoked products from cod species using enzyme preparation from hepatopancreas of snow crab Chionoecetes opilio

    Directory of Open Access Journals (Sweden)

    E. B. Shkuratova

    2017-01-01

    Full Text Available At the Food Production Department of Murmansk State Technical University (MSTU was developed a technology of production of smoked fish products with the use of air-flue mixture, produced in an infrared smoke generator under conditions of low-temperature pyrolysis of wood chips with an initial moisture content of 40 to 60% and a bulk density of from 104 to 154 kg/m3. A method of producing smoke and its hardware design enabled to securely control the temperature of the smoke receiving smoking environment, which guarantees a minimum content of threat to human health of polycyclic aromatic hydrocarbons (РАНs. The developed technology involves short cycle smoked salted semi – fish fillets and allows to obtain finished products with excellent organoleptic properties. The content of РАНs in finished products less than 0.0002 microgram/kg, which is significantly below the maximum permissible concentrations (MPC according to the requirements of SаnРiN 2.3.2.1078 (less than 0.001 mg/kg product and according to the requirements of TR CU 021/2011 (0.005 mg/kg product. This technology successfully implemented in production at fish processing plants in Murmansk region. However, marketing research has shown that the market for smoked fish in the Murmansk region is characterized by a narrow range, which affects consumer demand and reduces the competitiveness of the regional enterprises-manufacturers of smoked products. To solve this problem it is offered to expand the range of smoked fish products due to use of non-traditional Smoking techniques raw materials – fillet of cod fish (saithe, haddock, cod. To improve consumer properties of products, in particular, to improve the indicator of “consistency: and "juiciness" of the proposed use on the stage of the salting of prefabricated enzyme preparation from hepatopancreas of snow crab (Chionoecetes opilio, added to brine with density of 1.18 to 1.2 g/сm3 at a dosage of 0.04%, the curing time of

  20. Large propeptides of fungal β-N-acetylhexosaminidases are novel enzyme regulators that must be intracellularly processed to control activity, dimerization, and secretion into the extracellular environment

    Czech Academy of Sciences Publication Activity Database

    Plíhal, Ondřej; Sklenář, Jan; Hofbauerová, Kateřina; Novák, Petr; Man, Petr; Pompach, Petr; Kavan, Daniel; Ryšlavá, H.; Weignerová, Lenka; Charvátová, Andrea; Křen, Vladimír; Bezouška, Karel

    2007-01-01

    Roč. 46, - (2007), s. 2719-2734 ISSN 0006-2960 R&D Projects: GA MŠk LC545; GA MŠk LC06010; GA ČR GA203/04/1045; GA ČR GA203/05/0172; GA ČR GA204/06/0771; GA AV ČR IAA5020403 Institutional research plan: CEZ:AV0Z50200510 Keywords : enzyme regulator * filamentous fungi * hex Subject RIV: EE - Microbiology, Virology Impact factor: 3.368, year: 2007

  1. Antioxidant activity of protein hydrolysates from whole anchovy sprat (Clupeonella engrauliformis) prepared using endogenous enzymes and commercial proteases.

    Science.gov (United States)

    Ovissipour, Mahmoudreza; Rasco, Barbara; Shiroodi, Setareh Ghorban; Modanlow, Maryam; Gholami, Sanaz; Nemati, Mahrokh

    2013-05-01

    The antioxidant activity and chemical properties of fish protein hydrolysates (FPHs) prepared from anchovy sprat (Clupeonella engrauliformis) using endogenous enzymes (autolysis) and commercial proteases were investigated. The highest degree of hydrolysis (DH) was observed with Alcalase and papain and the highest protein recovery (PR) with Alcalase and bromelain. FPH yield was highest with Alcalase (82.3%) and lowest with autolysis (63.64%). Increased DH resulted in increased FPH yield (R(2) = 0.77). The highest oil recovery was observed with bromelain (6.41%) and the lowest with autolysis (3.58%). Antioxidant activity determined by DPPH, reducing power and ferrous chelation assays was highest in bromelain, Promod and papain FPHs respectively. The highest ABTS activity was observed in Alcalase FPH, followed by Promod and Protamex™ FPHs. The lowest antioxidant activity was observed in autolysed and Flavourzyme FPHs (P > 0.05). Heavy metals (arsenic, lead and mercury) were recorded at levels below the regulatory limits established by the FDA. Anchovy sprat hydrolysates showed high antioxidant activities and amino acid contents and low heavy metal concentrations, indicating that they have high potential for use in human and animal diets. The high antioxidant activities are related to the high levels of hydrophobic amino acids found in this study. © 2012 Society of Chemical Industry.

  2. Favorable effects of the detergent and enzyme extraction method for preparing decellularized bovine pericardium scaffold for tissue engineered heart valves.

    Science.gov (United States)

    Yang, Min; Chen, Chang-Zhi; Wang, Xue-Ning; Zhu, Ya-Bin; Gu, Y John

    2009-10-01

    Bovine pericardium has been extensively applied as the biomaterial for artificial heart valves and may potentially be used as a scaffold for tissue-engineered heart valves after decellularization. Although various methods of decellularization are currently available, it is unknown which method is the most ideal one for the decellularization for bovine pericardium. We compared three decellularization methods, namely, the detergent and enzyme extraction (DEE), the trypsin (TS), and the Triton X-100 and sodium-deoxycholate (TSD) method, to examine their efficacy on cell removal and their preservation of the mechanical function and the tissue matrix structure. Results indicated that decellularization was achieved by all the three methods as confirmed by hematoxylin-eosin staining, scanning electron microscopy, as well as quantitative DNA measurement. However, TS and TSD methods resulted in severe structural destruction of the bovine pericardium as shown by von Gieson staining and Gomori staining. Furthermore, both TS and TSD methods changed the mechanical property of the bovine pericardium, as evidenced by a lower elastic modulus, maximal-stress, maximal-disfiguration, maximal-load, and maximal-strain. In conclusion, the DEE method achieved both a complete decellularization and preservation of the mechanical function and tissue structure of the bovine pericardium. Thus, this method is superior to either the TS or the TSD method for preparing decellularized bovine pericardium scaffold for constructing tissue-engineered heart valves. (c) 2009 Wiley Periodicals, Inc.

  3. Preparation of anchovy (Engraulis japonicus) protein hydrolysates with high free radical-scavenging activity using endogenous and commercial enzymes.

    Science.gov (United States)

    He, Silian; Wang, Fanghua; Ning, Zhengxiang; Yang, Bo; Wang, Yonghua

    2014-12-01

    Anchovy protein hydrolysates with high free radical-scavenging activity were prepared by endogenous and commercial enzymes. Various hydrolytic factors (commercial protease composition, protease concentration, temperature, and reaction time) were optimized. Using a single-factor experiment, three commercial proteases (Protamex, Flavourzyme 500 MG, and Alcalase 2.4 L) were selected for further optimization using a simplex lattice design. The optimum composition of Protamex:Flavourzyme 500 MG:Alcalase 2.4 L was found to be 1.1:1.0:0.9. The hydrolytic conditions (commercial protease concentration, temperature, and reaction time) for the optimum protease composition were optimized using a Box-Behnken design. The optimum hydrolytic conditions were as follows: total commercial protease concentration of 3.27%, pH of 7.5, temperature of 55.4℃, and reaction time of 2.7 h. Under these conditions, hydrolysate with a 1, 1-diphenyl-2-picryhydrazyl scavenging activity of 84.7% was obtained. Meanwhile, a degree of hydrolysis of 33.2% and high protein nitrogen recovery of 87.5% were achieved. The amino acid composition of the hydrolysates demonstrated that they have high nutritional value, thereby suggesting that the hydrolysates have potential to be used as raw material for functional food. © The Author(s) 2013 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  4. Fungal diagnostics.

    Science.gov (United States)

    Kozel, Thomas R; Wickes, Brian

    2014-04-01

    Early diagnosis of fungal infection is critical to effective treatment. There are many impediments to diagnosis such as a diminishing number of clinical mycologists, cost, time to result, and requirements for sensitivity and specificity. In addition, fungal diagnostics must meet the contrasting needs presented by the increasing diversity of fungi found in association with the use of immunosuppressive agents in countries with high levels of medical care and the need for diagnostics in resource-limited countries where large numbers of opportunistic infections occur in patients with AIDS. Traditional approaches to diagnosis include direct microscopic examination of clinical samples, histopathology, culture, and serology. Emerging technologies include molecular diagnostics and antigen detection in clinical samples. Innovative new technologies that use molecular and immunoassay platforms have the potential to meet the needs of both resource-rich and resource-limited clinical environments.

  5. Oxidation of indole-3-acetic acid to oxindole-3-acetic acid by an enzyme preparation from Zea mays

    Science.gov (United States)

    Reinecke, D. M.; Bandurski, R. S.

    1988-01-01

    Indole-3-acetic acid is oxidized to oxindole-3-acetic acid by Zea mays tissue extracts. Shoot, root, and endosperm tissues have enzyme activities of 1 to 10 picomoles per hour per milligram protein. The enzyme is heat labile, is soluble, and requires oxygen for activity. Cofactors of mixed function oxygenase, peroxidase, and intermolecular dioxygenase are not stimulatory to enzymic activity. A heat-stable, detergent-extractable component from corn enhances enzyme activity 6- to 10-fold. This is the first demonstration of the in vitro enzymic oxidation of indole-3-acetic acid to oxindole-3-acetic acid in higher plants.

  6. Superficial fungal infections.

    Science.gov (United States)

    Kelly, Brendan P

    2012-04-01

    Tinea capitis, tinea corporis, and pityriasis versicolor are common superficial fungal infections in the pediatric population. • Tinea capitis is the most common dermatophyte infection worldwide. In North America, the cause is almost exclusively T tonsurans. Diagnosis of tinea capitis usually can be made by clinical features alone, especially when occipital or postauricular lymphadenopathy is present. Skin scrapings prepared with potassium hydroxide for microscopic examination, or a cotton swab for fungal culture, usually are diagnostic. • Treatment of tinea capitis requires systemic antifungal therapy. Terbinafine and griseofulvin are both effective against T tonsurans and are FDA-approved for this indication in children. • Adjunctive topical therapy for the patient and household contacts decreases transmission of this infection. • Topical antifungal therapy usually is effective for tinea corporis and pityriasis versicolor. However, recurrences of pityriasis versicolor are common.

  7. Production of Microbial Protease from Selected Soil Fungal Isolates ...

    African Journals Online (AJOL)

    Production of Microbial Protease from Selected Soil Fungal Isolates. ... Nigerian Journal of Biotechnology ... and 500C. The optimal pH on the enzyme production was observed to be between pH 3.5 and 5.5 for the organisms. Keywords: Soil microorganism, fungal isolate, incubation period, microbial enzyme. Nig J. Biotech.

  8. The fungal α-aminoadipate pathway for lysine biosynthesis requires two enzymes of the aconitase family for the isomerization of homocitrate to homoisocitrate.

    Science.gov (United States)

    Fazius, Felicitas; Shelest, Ekaterina; Gebhardt, Peter; Brock, Matthias

    2012-12-01

    Fungi produce α-aminoadipate, a precursor for penicillin and lysine via the α-aminoadipate pathway. Despite the biotechnological importance of this pathway, the essential isomerization of homocitrate via homoaconitate to homoisocitrate has hardly been studied. Therefore, we analysed the role of homoaconitases and aconitases in this isomerization. Although we confirmed an essential contribution of homoaconitases from Saccharomyces cerevisiae and Aspergillus fumigatus, these enzymes only catalysed the interconversion between homoaconitate and homoisocitrate. In contrast, aconitases from fungi and the thermophilic bacterium Thermus thermophilus converted homocitrate to homoaconitate. Additionally, a single aconitase appears essential for energy metabolism, glutamate and lysine biosynthesis in respirating filamentous fungi, but not in the fermenting yeast S. cerevisiae that possesses two contributing aconitases. While yeast Aco1p is essential for the citric acid cycle and, thus, for glutamate synthesis, Aco2p specifically and exclusively contributes to lysine biosynthesis. In contrast, Aco2p homologues present in filamentous fungi were transcribed, but enzymatically inactive, revealed no altered phenotype when deleted and did not complement yeast aconitase mutants. From these results we conclude that the essential requirement of filamentous fungi for respiration versus the preference of yeasts for fermentation may have directed the evolution of aconitases contributing to energy metabolism and lysine biosynthesis. © 2012 Blackwell Publishing Ltd.

  9. Effect of Enzyme Preparation with Activity Directed Towards Degradation of Non Starch Polysaccharides on Yellow Lupine Seed Based Diet for Young Broilers

    Directory of Open Access Journals (Sweden)

    Bogusław I Olkowski

    2010-01-01

    Full Text Available This work examined the impact of enzyme preparation with specific activity towards non starch polysaccharides on performance, morphological characteristics of gastrointestinal tract organs, microscopic evaluation of jejunal mucosa, and microbial status of ileum, caeca, and excreta in broilers fed a diet containing a high content of lupine meal. One-day-old chickens (Ross 308, mixed sex were randomly divided into control and experimental groups. Each group consisted of 36 birds, with 6 replications,and with 6 chickens per replication. The control group was fed the basal diet (consisting of maize and 40% of lupine, while the experimental treatment group was fed the basal diet supplemented with 0.06% commercial enzyme (Ronozyme VP. Chickens were fed diets in mash form for 4 weeks. Enzyme preparation significantly (P P P Enterobacteriaceae in caeca and excreta, and coliforms in excreta only (P < 0.01. Appropriate combination of enzyme preparations with activity towards degrading carbohydrates may offer a potential to reduce the deleterious impact of lupine in broilers.

  10. Fungal type III polyketide synthases.

    Science.gov (United States)

    Hashimoto, Makoto; Nonaka, Takamasa; Fujii, Isao

    2014-10-01

    This article covers the literature on fungal type III polyketide synthases (PKSs) published from 2005 to 2014. Since the first discovery of fungal type III PKS genes in Aspergillus oryzae, reported in 2005, putative genes for type III PKSs have been discovered in fungal genomes. Compared with type I PKSs, type III PKSs are much less abundant in fungi. However, type III PKSs could have some critical roles in fungi. This article summarizes the studies on fungal type III PKS functional analysis, including Neurospora crassa ORAS, Aspergillus niger AnPKS, Botrytis cinerea BPKS and Aspergillus oryzae CsyA and CsyB. It is mostly in vitro analysis using their recombinant enzymes that has revealed their starter and product specificities. Of these, CsyB was found to be a new kind of type III PKS that catalyses the coupling of two β-keto fatty acyl CoAs. Homology modelling reported in this article supports the importance of the capacity of the acyl binding tunnel and active site cavity in fungal type III PKSs.

  11. Enzyme Informatics

    Science.gov (United States)

    Alderson, Rosanna G.; Ferrari, Luna De; Mavridis, Lazaros; McDonagh, James L.; Mitchell, John B. O.; Nath, Neetika

    2012-01-01

    Over the last 50 years, sequencing, structural biology and bioinformatics have completely revolutionised biomolecular science, with millions of sequences and tens of thousands of three dimensional structures becoming available. The bioinformatics of enzymes is well served by, mostly free, online databases. BRENDA describes the chemistry, substrate specificity, kinetics, preparation and biological sources of enzymes, while KEGG is valuable for understanding enzymes and metabolic pathways. EzCatDB, SFLD and MACiE are key repositories for data on the chemical mechanisms by which enzymes operate. At the current rate of genome sequencing and manual annotation, human curation will never finish the functional annotation of the ever-expanding list of known enzymes. Hence there is an increasing need for automated annotation, though it is not yet widespread for enzyme data. In contrast, functional ontologies such as the Gene Ontology already profit from automation. Despite our growing understanding of enzyme structure and dynamics, we are only beginning to be able to design novel enzymes. One can now begin to trace the functional evolution of enzymes using phylogenetics. The ability of enzymes to perform secondary functions, albeit relatively inefficiently, gives clues as to how enzyme function evolves. Substrate promiscuity in enzymes is one example of imperfect specificity in protein-ligand interactions. Similarly, most drugs bind to more than one protein target. This may sometimes result in helpful polypharmacology as a drug modulates plural targets, but also often leads to adverse side-effects. Many cheminformatics approaches can be used to model the interactions between druglike molecules and proteins in silico. We can even use quantum chemical techniques like DFT and QM/MM to compute the structural and energetic course of enzyme catalysed chemical reaction mechanisms, including a full description of bond making and breaking. PMID:23116471

  12. Preparation

    Directory of Open Access Journals (Sweden)

    M.M. Dardir

    2014-03-01

    Full Text Available Some hexanamide-mono and di-linoleniate esters were prepared by the reaction of linolenic acid and hexanamide (derived from the reaction of hexanoic acid and diethanolamine. The chemical structure for the newly prepared hexanamide-mono and di-linoleniate esters were elucidated using elemental analysis, (FTIR, H 1NMR and chemical ionization mass spectra (CI/Ms spectroscopic techniques. The results of the spectroscopic analysis indicated that they were prepared through the right method and they have high purity. The new prepared esters have high biodegradability and lower toxicity (environmentally friendly so they were evaluated as a synthetic-based mud (ester-based mud for oil-well drilling fluids. The evaluation included study of the rheological properties, filtration and thermal properties of the ester based-muds formulated with the newly prepared esters compared to the reference commercial synthetic-based mud.

  13. The effects of preparing methods and enzyme supplementation on the utilization of brown marine algae (Sargassum dentifebium meal in the diet of laying hens

    Directory of Open Access Journals (Sweden)

    Mohammed A. Al-Harthi

    2011-10-01

    Full Text Available Brown marine algae (BMA; Sargassum dentifebium were collected from Jeddah on the shores of the Red Sea and sun dried at an average daily temperature of 40°C until constant weight was obtained. Part of the sun dried brown marine algae was subsequently processed by boiling (BBMA;boiled brown marine algae in water and by autoclaving (ABMA; autoclaved brown marine algae. The SBMA, BBMA and ABMA were included in laying hen diet during weeks 23-42 of age at concentrations of 0.0%, 3.0% and 6.0%. The diets were given with or without enzyme supplementation. This resulted in 3 (preparation methods × 2 (concentrations of supplemented BMA, i.e. 3 and 6 % × 2 (with and without enzyme supplementation diet programs plus two control groups (with and without enzyme supplementation for a total of 14 treatments. Each treatment was represented by six replicates of five hens each. Sun dried or autocalved brown marine algae at 3% without enzyme supplementation in the laying hen diet could be fed to laying hens without any adverse effect on laying performance. However, enzyme supplementation to a diet containing 6% autocalved brown marine algae improved productive performance and eggshell quality.

  14. Soil fungal community responses to global changes

    DEFF Research Database (Denmark)

    Haugwitz, Merian Skouw

    Global change will affect the functioning and structure of terrestrial ecosystems and since soil fungi are key players in organic matter decomposition and nutrient turnover, shifts in fungal community composition might have a strong impact on soil functioning. The main focus of this thesis...... was therefore to investigate the impact of global environmental changes on soil fungal communities in a temperate and subartic heath ecosystem. The objective was further to determine global change effects on major functional groups of fungi and analyze the influence of fungal community changes on soil carbon...... and nutrient availability and storage. By combining molecular methods such as 454 pyrosequencing and quantitative PCR of fungal ITS amplicons with analyses of soil enzymes, nutrient pools of carbon, nitrogen and phosphorus we were able to characterize soil fungal communities as well as their impact on nutrient...

  15. Cytochrome P450 enzyme systems in fungi

    NARCIS (Netherlands)

    Brink, H.M. van den; Gorcom, R.F.M. van; Hondel, C.A.M.J.J. van den; Punt, P.J.

    1998-01-01

    The involvement of cytochrome P450 enzymes in many complex fungal bioconversion processes has been characterized in recent years. Accordingly, there is now considerable scientific interest in fungal cytochrome P450 enzyme systems. In contrast to S. cerevisiae, where surprisingly few P450 genes have

  16. Directed Evolution of Fungal Laccases

    OpenAIRE

    Maté, Diana; García-Ruiz, Eva; Camarero, Susana; Alcalde, Miguel

    2011-01-01

    Fungal laccases are generalists biocatalysts with potential applications that range from bioremediation to novel green processes. Fuelled by molecular oxygen, these enzymes can act on dozens of molecules of different chemical nature, and with the help of redox mediators, their spectrum of oxidizable substrates is further pushed towards xenobiotic compounds (pesticides, industrial dyes, PAHs), biopolymers (lignin, starch, cellulose) and other complex molecules. In recent years, extraordinary e...

  17. Genetic engineering of crop plants for fungal resistance: role of antifungal genes.

    Science.gov (United States)

    Ceasar, S Antony; Ignacimuthu, S

    2012-06-01

    Fungal diseases damage crop plants and affect agricultural production. Transgenic plants have been produced by inserting antifungal genes to confer resistance against fungal pathogens. Genes of fungal cell wall-degrading enzymes, such as chitinase and glucanase, are frequently used to produce fungal-resistant transgenic crop plants. In this review, we summarize the details of various transformation studies to develop fungal resistance in crop plants.

  18. Preparation of Stable Cross-Linked Enzyme Aggregates (CLEAs of a Ureibacillus thermosphaericus Esterase for Application in Malathion Removal from Wastewater

    Directory of Open Access Journals (Sweden)

    Yuliya V. Samoylova

    2018-04-01

    Full Text Available In this study, the active and stable cross-linked enzyme aggregates (CLEAs of the thermostable esterase estUT1 of the bacterium Ureibacillus thermosphaericus were prepared for application in malathion removal from municipal wastewater. Co-expression of esterase with an E. coli chaperone team (KJE, ClpB, and ELS increased the activity of the soluble enzyme fraction up to 200.7 ± 15.5 U mg−1. Response surface methodology (RSM was used to optimize the preparation of the CLEA-estUT1 biocatalyst to maximize its activity and minimize enzyme loss. CLEA-estUT1 with the highest activity of 29.4 ± 0.5 U mg−1 (90.6 ± 2.7% of the recovered activity was prepared with 65.1% (w/v ammonium sulfate, 120.6 mM glutaraldehyde, and 0.2 mM bovine serum albumin at 5.1 h of cross-linking. The biocatalyst has maximal activity at 80 °С and pH 8.0. Analysis of the properties of CLEA-estUT1 and free enzyme at 50–80 °C and pH 5.0–10.0 showed higher stability of the biocatalyst. CLEA-estUT1 showed marked tolerance against a number of chemicals and high operational stability and activity in the reaction of malathion hydrolysis in wastewater (up to 99.5 ± 1.4%. After 25 cycles of malathion hydrolysis at 37 °С, it retained 55.2 ± 1.1% of the initial activity. The high stability and reusability of CLEA-estUT1 make it applicable for the degradation of insecticides.

  19. Fungal enzymes in the attine ant symbiosis

    DEFF Research Database (Denmark)

    de Fine Licht, Henrik Hjarvard; Schiøtt, Morten; Boomsma, Jacobus Jan

    build huge nests displacing several cubic meters of soil, whereas lower attine genera such as Cyphomyrmex and Mycocepurus have small nests with a fungus garden the size of a table-tennis ball. Only the leaf-cutter ants are specialized on using fresh leaves as substrate for their fungus gardens, whereas...

  20. Comparison of a pectinolytic extract of Kluyveromyces marxianus and a commercial enzyme preparation in the production of Ives (Vitis labrusca) grape juice.

    Science.gov (United States)

    Piemolini-Barreto, Luciani Tatsch; Antônio, Regina Vasconcellos; Echeverrigaray, Sergio

    2015-05-01

    This study analyses the effect of the crude enzymatic extract produced by Kluyveromyces marxianus (EEB) in the maceration and clarification of juice produced from Ives (Vitis labrusca) grapes compared to the commercial enzyme preparation Pectinex(®)Ultra Color (PEC). Treatments were conducted with a total pectinolytic activity of 1 U/mL of fruit juice, at 40 °C, for 60 min. After the enzymatic treatment, the juices were evaluated with respect to yield, viscosity, and degree of clarification, as well as the effect of the enzymes on polyphenol concentration, anthocyanins, and juice color. The results showed that both EEB and PEC increase yield, reduce viscosity and contribute to the clarification of grape juice. After enzyme treatment with the EEB preparation, the extraction yield increased 28.02 % and decreased 50.70 % in viscosity during the maceration of the pulp. During the juice production process clarification increased 11.91 %. With PEC, higher values for these parameters: 42.36, 63.20, and 26.81 % respectively, were achieved. The addition of EEB resulted in grape juice with better color intensity and extraction of phenolic compounds and anthocyanins. Considering all comparison criteria, the enzymatic extract of K. marxianus NRRL-Y-7571 can potentially be used in the production of juice.

  1. [A comparative characterization of fungal melanin and the humin-like substances synthesized by Cerrena maxima 0275].

    Science.gov (United States)

    Koroleva, O V; Kulikova, N A; Alekseeva, T N; Stepanova, E V; Davidchik, V N; Beliaeva, E Iu; Tsvetkova, E A

    2007-01-01

    Comparative studies of fungal melanin and two preparations of the high-molecular-weight humin-like substances formed during a solid-phase cultivation of the basidiomycete Cerrena maxima 0275 for 45 and 70 days were performed. The fungal melanin from Aspergillus niger and the humin-like substances synthesized by the basidiomycete C. maxima 0275 are similar in their physicochemical properties (elemental composition and behavior in acids and alkalis) and auxin-like activities. However, these biopolymers differ, essentially, at the structural level. According to IR spectroscopy data, the obtained humin-like substances display a higher similarity to natural humic acids and are more diverse in their functional groups compared with fungal melanins. Presumably, this is connected with the fact that laccase is involved in formation of humin-like substances; moreover, this enzyme is involved not only in the synthesis of these polymers, but also in their modification and degradation.

  2. Synthesis of 5'-oligonucleotide hydrazide derivatives and their use in preparation of enzyme-nucleic acid hybridization probes.

    Science.gov (United States)

    Ghosh, S S; Kao, P M; Kwoh, D Y

    1989-04-01

    A hydrazone-based method for conjugating synthetic nucleic acids and reporter molecules for use as nonradioactive hybridization probes is presented. Oligonucleotides complementary to the hepatitis B virus were derivatized at their 5' ends with hydrazine or homobifunctional acyl hydrazides. These derivatives reacted facilely with aldehydes to give hydrazones, which were characterized by uv spectroscopy and HPLC. Coupling of aldehyde-modified alkaline phosphatase with carbohydrazide-oligonucleotide derivatives provided a mixture of two enzyme-nucleic acid conjugates in 80-85% yield. The conjugates had a 1:1 and a 2:1 oligonucleotide/enzyme ratio, respectively, and were separated by ion-exchange chromatography. Both conjugates were able to detect 7 amol of target DNA in 1 h, using a colorimetric assay. In contrast, oligonucleotide-horseradish peroxidase conjugates were 40-fold lower in sensitivity of detection.

  3. Optimisation of synergistic biomass-degrading enzyme systems for efficient rice straw hydrolysis using an experimental mixture design.

    Science.gov (United States)

    Suwannarangsee, Surisa; Bunterngsook, Benjarat; Arnthong, Jantima; Paemanee, Atchara; Thamchaipenet, Arinthip; Eurwilaichitr, Lily; Laosiripojana, Navadol; Champreda, Verawat

    2012-09-01

    Synergistic enzyme system for the hydrolysis of alkali-pretreated rice straw was optimised based on the synergy of crude fungal enzyme extracts with a commercial cellulase (Celluclast™). Among 13 enzyme extracts, the enzyme preparation from Aspergillus aculeatus BCC 199 exhibited the highest level of synergy with Celluclast™. This synergy was based on the complementary cellulolytic and hemicellulolytic activities of the BCC 199 enzyme extract. A mixture design was used to optimise the ternary enzyme complex based on the synergistic enzyme mixture with Bacillus subtilis expansin. Using the full cubic model, the optimal formulation of the enzyme mixture was predicted to the percentage of Celluclast™: BCC 199: expansin=41.4:37.0:21.6, which produced 769 mg reducing sugar/g biomass using 2.82 FPU/g enzymes. This work demonstrated the use of a systematic approach for the design and optimisation of a synergistic enzyme mixture of fungal enzymes and expansin for lignocellulosic degradation. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Preparation of well-dispersed gold/magnetite nanoparticles embedded on cellulose nanocrystals for efficient immobilization of papain enzyme.

    Science.gov (United States)

    Mahmoud, Khaled A; Lam, Edmond; Hrapovic, Sabahudin; Luong, John H T

    2013-06-12

    A nanocomposite consisting of magnetite nanoparticles (Fe3O4NPs) and Au nanoparticles (AuNPs) embedded on cellulose nanocrystals (CNCs) was used as a magnetic support for the covalent conjugation of papain and facilitated recovery of this immobilized enzyme. Fe3O4NPs (10-20 nm in diameter) and AuNPs (3-7 nm in diameter) were stable and well-dispersed on the CNC surface. Energy-dispersive spectroscopy, X-ray diffraction, and Fourier transform infrared spectroscopy were used to evaluate the surface composition and structure of CNC/Fe3O4NPs/AuNPs. The nanocomposite was successfully used for the immobilization and separation of papain from the reaction mixture. The optimal enzyme loading was 186 mg protein/g CNC/Fe3O4NPs/AuNPs, significantly higher than the value reported in the literature. The activity of immobilized papain was studied by electrochemical detection of its specific binding to the Thc-Fca-Gly-Gly-Tyr-Arg inhibitory sequence bound to an Au electrode. The immobilized enzyme retained 95% of its initial activity after 35 days of storage at 4 °C, compared to 41% for its free form counterpart.

  5. Chlorination and cleavage of lignin structures by fungal chloroperoxidases

    Science.gov (United States)

    Patricia Ortiz-Bermudez; Ewald Srebotnik; Kenneth E. Hammel

    2003-01-01

    Two fungal chloroperoxidases (CPOs), the heme enzyme from Caldariomyces fumago and the vanadium enzyme from Curvularia inaequalis, chlorinated 1-(4-ethoxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)-1,3-dihydroxypropane, a dimeric model compound that represents the major nonphenolic structure in lignin. Both enzymes also cleaved this dimer to give 1-chloro-4-ethoxy-3-...

  6. Structure and biosynthesis of fungal alpha-glucans

    NARCIS (Netherlands)

    Grün, Christian Hugo

    2003-01-01

    The fungal cell wall is unique among eukaryotes and therefore it forms an ideal target for the development of novel antifungal drugs. Fungal cell morphology and integrity depend on a cell-surrounding wall, which is composed of glycoproteins and polysaccharides. Disrupting enzymes that are involved

  7. Fungal inhibitory lactic acid bacteria

    OpenAIRE

    Ström, Katrin

    2005-01-01

    Lactic acid bacteria (LAB) are microorganisms that have been used for centuries to prepare and improve storage of food and for ensiling of different crops for animal feed. This thesis explores the possibility of using LAB to inhibit growth of spoilage fungi in food and feed products. LAB isolates, collected from plant material or dairy products, were screened for antifungal activity in a dual culture assay. Strains with antifungal activity were identified and the fungal inhibitory activity wa...

  8. Soil fungal community responses to global changes

    DEFF Research Database (Denmark)

    Haugwitz, Merian Skouw

    and nutrient availability and storage. By combining molecular methods such as 454 pyrosequencing and quantitative PCR of fungal ITS amplicons with analyses of soil enzymes, nutrient pools of carbon, nitrogen and phosphorus we were able to characterize soil fungal communities as well as their impact on nutrient...... storage in both a shorter and longer term global change experiment. Extended drought at the dry, temperate heath showed that soil fungi were well adapted to dry conditions. Furthermore, soil fungal communities responded significantly to seasonal fluctuations at the temperate heath, but despite large......Global change will affect the functioning and structure of terrestrial ecosystems and since soil fungi are key players in organic matter decomposition and nutrient turnover, shifts in fungal community composition might have a strong impact on soil functioning. The main focus of this thesis...

  9. Improving the direct electron transfer in monolithic bioelectrodes prepared by immobilization of FDH enzyme on carbon-coated anodic aluminum oxide films

    Directory of Open Access Journals (Sweden)

    Alberto eCastro-Muñiz

    2016-02-01

    Full Text Available The present work reports the preparation of binderless carbon-coated porous films and the study of their performance as monolithic bioanodes. The films were prepared by coating anodic aluminum oxide (AAO films with a thin layer of nitrogen-doped carbon by chemical vapor deposition. The films have cylindrical straight pores with controllable diameter and length. These monolithic films were used directly as bioelectrodes by loading the films with D-fructose dehydrogenase (FDH, an oxidoreductase enzyme that catalyzes the oxidation of D-fructose to 5-keto-D-fructose. The immobilization of the enzymes was carried out by physical adsorption in liquid phase and with an electrostatic attraction method. The latter method takes advantage of the fact that FDH is negatively charged during the catalytic oxidation of fructose. Thus the immobilization was performed under the application of a positive voltage to the CAAO film in a FDH-fructose solution in McIlvaine buffer (pH 5 at 25 ºC. As a result, the FDH modified electrodes with the latter method show much better electrochemical response than that with the conventional physical adsorption method. Due to the singular porous structure of the monolithic films, which consists of an array of straight and parallel nanochannels, it is possible to rule out the effect of the diffusion of the D-fructose into the pores. Thus the improvement in the performance upon using the electrostatic attraction method can be ascribed not only to a higher uptake, but also to a more appropriate molecule orientation of the enzyme units on the surface of the electrodes.

  10. Role of fungal peroxidases in biological ligninolysis

    Science.gov (United States)

    Kenneth E. Hammel; Dan Cullen

    2008-01-01

    The degradation of lignin by filamentous fungi is a major route for the recycling of photosynthetically fixed carbon, and the oxidative mechanisms employed have potential biotechnological applications. The lignin peroxidases (LiPs), manganese peroxidases (MnPs), and closely related enzymes of white rot basidiomycetes are likely contributors to fungal ligninolysis. Many...

  11. Fungal cultivation on glass-beads

    DEFF Research Database (Denmark)

    Droce, Aida; Sørensen, Jens Laurids; Giese, Henriette

    Transcription of various bioactive compounds and enzymes are dependent on fungal cultivation method. In this study we cultivate Fusarium graminearum and Fusarium solani on glass-beads with liquid media in petri dishes as an easy and inexpensive cultivation method, that resembles in secondary...

  12. A biotechnology perspective of fungal proteases

    Directory of Open Access Journals (Sweden)

    Paula Monteiro de Souza

    2015-06-01

    Full Text Available Proteases hydrolyze the peptide bonds of proteins into peptides and amino acids, being found in all living organisms, and are essential for cell growth and differentiation. Proteolytic enzymes have potential application in a wide number of industrial processes such as food, laundry detergent and pharmaceutical. Proteases from microbial sources have dominated applications in industrial sectors. Fungal proteases are used for hydrolyzing protein and other components of soy beans and wheat in soy sauce production. Proteases can be produced in large quantities in a short time by established methods of fermentation. The parameters such as variation in C/N ratio, presence of some sugars, besides several other physical factors are important in the development of fermentation process. Proteases of fungal origin can be produced cost effectively, have an advantage faster production, the ease with which the enzymes can be modified and mycelium can be easily removed by filtration. The production of proteases has been carried out using submerged fermentation, but conditions in solid state fermentation lead to several potential advantages for the production of fungal enzymes. This review focuses on the production of fungal proteases, their distribution, structural-functional aspects, physical and chemical parameters, and the use of these enzymes in industrial applications.

  13. A biotechnology perspective of fungal proteases

    Science.gov (United States)

    de Souza, Paula Monteiro; Bittencourt, Mona Lisa de Assis; Caprara, Carolina Canielles; de Freitas, Marcela; de Almeida, Renata Paula Coppini; Silveira, Dâmaris; Fonseca, Yris Maria; Ferreira, Edivaldo Ximenes; Pessoa, Adalberto; Magalhães, Pérola Oliveira

    2015-01-01

    Proteases hydrolyze the peptide bonds of proteins into peptides and amino acids, being found in all living organisms, and are essential for cell growth and differentiation. Proteolytic enzymes have potential application in a wide number of industrial processes such as food, laundry detergent and pharmaceutical. Proteases from microbial sources have dominated applications in industrial sectors. Fungal proteases are used for hydrolyzing protein and other components of soy beans and wheat in soy sauce production. Proteases can be produced in large quantities in a short time by established methods of fermentation. The parameters such as variation in C/N ratio, presence of some sugars, besides several other physical factors are important in the development of fermentation process. Proteases of fungal origin can be produced cost effectively, have an advantage faster production, the ease with which the enzymes can be modified and mycelium can be easily removed by filtration. The production of proteases has been carried out using submerged fermentation, but conditions in solid state fermentation lead to several potential advantages for the production of fungal enzymes. This review focuses on the production of fungal proteases, their distribution, structural-functional aspects, physical and chemical parameters, and the use of these enzymes in industrial applications. PMID:26273247

  14. Fungal infections of the oral cavity.

    Science.gov (United States)

    Zegarelli, D J

    1993-12-01

    Although several strains of Candida can infect the oral mucosa, the most commonly encountered oral fungal infection is Candida albicans, which may be highly infective because of its greater level of pathogenicity and adherence properties. C. albicans is an oral commensal in as many as 40% to 65% of healthy adult mouths. The papillated dorsal surface of the tongue and palatal mucosa beneath a maxillary denture are favored reservoir sites. Oral candidal infection almost always involves a compromised host. The compromise may be local or systemic. Local factors include decreased salivation and the weaning of dentures. Systemic factors include diabetes mellitus, pernicious anemia, and AIDS. Some have even implicated advanced age and the female gender as being mild predisposing factors. Furthermore, the C. albicans infection itself can depress a host's immune system. A patient with oral candidiasis can present with one or more of the following clinical forms: pseudomembranous, erythematous, hyperplastic, and denture erythematous. Many investigators accept median rhomboid glossitis as a form of chronic oral candidiasis. In some patients with angular cheilitis, genesis of the lesions is secondary to monilial infestation. Because C. albicans is a normal inhabitant in many mouths, diagnostic confirmation of infection often rests with successful response (i.e., resolution of lesions) to antifungal medications. This form of diagnostic confirmation can be further enhanced by culturing the offending microbe, preparing a fungal smear, or even incisional biopsy. The microscopic demonstration of fungal hyphae is highly diagnostic of the candidal infection, whether the hyphae are demonstrated on a PAS smear or on a biopsy within surface stratified squamous epithelium. Numerous medications exist for the treatment of oral candidiasis. They include the antibiotic nystatin as well as clotrimazole, ketoconazole, and fluconazole. Nystatin is safe and is used as a topical agent in rinse or

  15. Etiological Analysis of Fungal Keratitis and Rapid Identification of Predominant Fungal Pathogens.

    Science.gov (United States)

    He, Dan; Hao, Jilong; Gao, Song; Wan, Xue; Wang, Wanting; Shan, Qiushi; Wang, Li

    2016-02-01

    Fungal keratitis is a worldwide-distributed refractory and potentially blinding ocular infection caused by various fungi. It is necessary to investigate the etiological and epidemiological characteristics of this disease and establish a rapid and specific pathogenic identification method. Here, we isolated and identified fungal pathogens of 275 patients with presumed fungal keratitis from Jilin Province, China, and conducted statistical analyses of epidemiological information. The positive rate of fungal culture was 72.0 %. Fusarium sp. was the most common genus among 210 fungal isolates. The predominant species were Fusarium solani, Aspergillus fumigatus, and Candida glabrata, which accounted for over 50 % of the isolated organisms. Corneal trauma and previous use of drugs were the most important predisposing factors. In addition, a multiplex polymerase chain reaction (PCR) was designed with species-specific primers of the three species that could identify them with amplicons of approximately 330 bp from F. solani, 275 bp from A. fumigatus, and 230 bp from C. glabrata. Additionally, PCR with fungal universal primers and multiplex PCR were performed using DNA prepared by an improved DNA extraction method from corneal scrapings. With this method, fungal pathogens from corneal scrapings could be specifically and rapidly identified within 8 h. The culture-independent rapid identification of corneal scrapings may have great significance for the early diagnosis and treatment of fungal keratitis.

  16. The effect of commercial enzyme preparation-assisted maceration on the yield, quality, and bioactivity of essential oil from waste carrot seeds (Daucus carota L.

    Directory of Open Access Journals (Sweden)

    Śmigielski, K. B.

    2014-12-01

    Full Text Available Eight enzyme preparations were screened with a view to maximizing the yield of carrot seed essential oil. Three of the eight enzyme preparations investigated, lipase from Mucor circinelloides, XPect® pectinase, and Esperase® protease, significantly influenced the amount of essential oil obtained, with Esperase® being the most effective. The Taguchi method was applied to optimize the processing conditions for the Esperase® protease. Under the optimum conditions, the essential oil yield increased by approximately 48%. The main constituent compounds in the oil are: carotol (OeA: 40.80%–OeB: 46.17%, daucol (OeA: 7.35%–OeB: 6.22%, sabinene (OeA: 5.12%–OeB: 6.13%, alpha-pinene (OeA: 4.24%–OeB: 5.11% and geranyl acetate (OeA: 4.50%–OeB: 3.68%. As compared to the control sample, the essential oil obtained from enzyme-pretreated carrot seeds has the same biological activity against Bacillus subtilis and Candida sp., lower activity against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, and higher activity against Aspergillus niger and Penicillium expansum.Ocho preparados enzimáticos fueron seleccionados con el fin de maximizar el rendimiento de aceites esenciales de semillas de zanahoria. Tres de los ocho preparados de las enzimas investigadas, lipasa de Mucor circinelloides, Xpect® pectinasa y Esperase® proteasa, influyeron de manera significativa sobre la cantidad de aceite esencial obtenido, siendo Esperase® el más eficaz. El método de Taguchi se aplicó para optimizar las condiciones del procesamiento para esta última. Bajo las condiciones óptimas, el rendimiento de los aceite esenciales aumentó aproximadamente un 48%. Los principales compuestos constituyentes del aceite son: carotol (OEA: 40.80%–OeB: 46,17%, ducol (OEA: 7,35%–OeB: 6,22%, sabineno (OEA: 5,12%–OeB: 6,13%, alfa-pineno (OEA: 4,24%– OeB: 5,11% y acetato de geranilo (OEA: 4,50%–OeB: 3,68%. En comparación con la muestra control, el

  17. Enzyme and biochemical producing fungi

    DEFF Research Database (Denmark)

    Lübeck, Peter Stephensen; Lübeck, Mette; Nilsson, Lena

    2010-01-01

    We are developing a biorefinery concept for biological production of chemicals, drugs, feed and fuels using plant biomass as raw material in well-defined cell-factories. Among the important goals is the discovery of new biocatalysts for production of enzymes, biochemicals and fuels and already our...... screening of a large collection of fungal strains isolated from natural habitats have resulted in identification of strains with high production of hydrolytic enzymes and excretion of organic acids. Our research focuses on creating a fungal platform based on synthetic biology for developing new cell...

  18. Anaerobic fungal populations

    International Nuclear Information System (INIS)

    Brookman, J.L.; Nicholson, M.J.

    2005-01-01

    The development of molecular techniques has greatly broadened our view of microbial diversity and enabled a more complete detection and description of microbial communities. The application of these techniques provides a simple means of following community changes, for example, Ishii et al. described transient and more stable inhabitants in another dynamic microbial system, compost. Our present knowledge of anaerobic gut fungal population diversity within the gastrointestinal tract is based upon isolation, cultivation and observations in vivo. It is likely that there are many species yet to be described, some of which may be non-culturable. We have observed a distinct difference in the ease of cultivation between the different genera, for example, Caecomyes isolates are especially difficult to isolate and maintain in vitro, a feature that is likely to result in the under representation of this genera in culture-based enumerations. The anaerobic gut fungi are the only known obligately anaerobic fungi. For the majority of their life cycles, they are found tightly associated with solid digesta in the rumen and/or hindgut. They produce potent fibrolytic enzymes and grow invasively on and into the plant material they are digesting making them important contributors to fibre digestion. This close association with intestinal digesta has made it difficult to accurately determine the amount of fungal biomass present in the rumen, with Orpin suggesting 8% contribution to the total microbial biomass, whereas Rezaeian et al. more recently gave a value of approximately 20%. It is clear that the rumen microbial complement is affected by dietary changes, and that the fungi are more important in digestion in the rumens of animals fed with high-fibre diets. It seems likely that the gut fungi play an important role within the rumen as primary colonizers of plant fibre, and so we are particularly interested in being able to measure the appearance and diversity of fungi on the plant

  19. Freshwater Fungal Infections

    Directory of Open Access Journals (Sweden)

    Dennis J. Baumgardner

    2017-01-01

    Full Text Available Fungal infections as a result of freshwater exposure or trauma are fortunately rare. Etiologic agents are varied, but commonly include filamentous fungi and Candida. This narrative review describes various sources of potential freshwater fungal exposure and the diseases that may result, including fungal keratitis, acute otitis externa and tinea pedis, as well as rare deep soft tissue or bone infections and pulmonary or central nervous system infections following traumatic freshwater exposure during natural disasters or near-drowning episodes. Fungal etiology should be suspected in appropriate scenarios when bacterial cultures or molecular tests are normal or when the infection worsens or fails to resolve with appropriate antibacterial therapy.

  20. Spatial and temporal control of fungal natural product synthesis.

    Science.gov (United States)

    Lim, Fang Yun; Keller, Nancy P

    2014-10-01

    Despite their oftentimes-elusive ecological role, fungal natural products have, for better or worse, impacted our daily lives tremendously owing to their diverse and potent bioactive properties. This Janus-faced nature of fungal natural products inevitably ushered in a field of research dedicated towards understanding the ecology, organisms, genes, enzymes, and biosynthetic pathways that give rise to this arsenal of diverse and complex chemistry. Ongoing research in fungal secondary metabolism has not only increased our appreciation for fungal natural products as an asset but also sheds light on the pivotal role that these once-regarded "metabolic wastes" play in fungal biology, defense, and stress response in addition to their potential contributions towards human mycoses. Full orchestration of secondary metabolism requires not only the seamless coordination between temporal and spatial control of SM-associated machineries (e.g. enzymes, cofactors, intermediates, and end-products) but also integration of these machineries into primary metabolic processes and established cellular mechanisms. An intriguing, but little known aspect of microbial natural product synthesis lies in the spatial organization of both pathway intermediates and enzymes responsible for the production of these compounds. In this highlight, we summarize some major breakthroughs in understanding the genes and regulation of fungal natural product synthesis and introduce the current state of knowledge on the spatial and temporal control of fungal natural product synthesis.

  1. Versatile biocatalysis of fungal cytochrome P450 monooxygenases.

    Science.gov (United States)

    Durairaj, Pradeepraj; Hur, Jae-Seoun; Yun, Hyungdon

    2016-07-18

    Cytochrome P450 (CYP) monooxygenases, the nature's most versatile biological catalysts have unique ability to catalyse regio-, chemo-, and stereospecific oxidation of a wide range of substrates under mild reaction conditions, thereby addressing a significant challenge in chemocatalysis. Though CYP enzymes are ubiquitous in all biological kingdoms, the divergence of CYPs in fungal kingdom is manifold. The CYP enzymes play pivotal roles in various fungal metabolisms starting from housekeeping biochemical reactions, detoxification of chemicals, and adaptation to hostile surroundings. Considering the versatile catalytic potentials, fungal CYPs has gained wide range of attraction among researchers and various remarkable strategies have been accomplished to enhance their biocatalytic properties. Numerous fungal CYPs with multispecialty features have been identified and the number of characterized fungal CYPs is constantly increasing. Literature reveals ample reviews on mammalian, plant and bacterial CYPs, however, modest reports on fungal CYPs urges a comprehensive review highlighting their novel catalytic potentials and functional significances. In this review, we focus on the diversification and functional diversity of fungal CYPs and recapitulate their unique and versatile biocatalytic properties. As such, this review emphasizes the crucial issues of fungal CYP systems, and the factors influencing efficient biocatalysis.

  2. Enzymic lactose hydrolysis

    Energy Technology Data Exchange (ETDEWEB)

    Miller, J.J.; Brand, J.C.

    1980-01-01

    Acid or enzymic hydrolysis can be used to hydrolyze lactose. Advantages of both are compared and details of enzymic hydrolysis using yeast or fungal enzymes given. The new scheme outlined involves recycling lactase. Because lactose and lactase react to ultrafiltration (UF) membranes differently separation is possible. Milk or milk products are ultrafiltered to separate a concentrate from a lactose-rich permeate which is treated with lactase in a reactor until hydrolysis reaches a required level. The lactase can be removed by UF as it does not permeate the membrane, and it is recycled back to the reactor. Permeate from the second UF stage may or may not be recombined with the concentrate from the first stage to produce a low lactose product (analysis of a typical low-lactose dried whole milk is given). Batch or continuous processes are explained and a batch process without enzyme recovery is discussed. (Refs. 4).

  3. A parts list for fungal cellulosomes revealed by comparative genomics

    Energy Technology Data Exchange (ETDEWEB)

    Haitjema, Charles H.; Gilmore, Sean P.; Henske, John K.; Solomon, Kevin V.; de Groot, Randall; Kuo, Alan; Mondo, Stephen J.; Salamov, Asaf A.; LaButti, Kurt; Zhao, Zhiying; Chiniquy, Jennifer; Barry, Kerrie; Brewer, Heather M.; Purvine, Samuel O.; Wright, Aaron T.; Hainaut, Matthieu; Boxma, Brigitte; van Alen, Theo; Hackstein, Johannes H. P.; Henrissat, Bernard; Baker, Scott E.; Grigoriev, Igor V.; O' Malley, Michelle A.

    2017-05-26

    Cellulosomes are large, multi-protein complexes that tether plant biomass degrading enzymes together for improved hydrolysis1. These complexes were first described in anaerobic bacteria where species specific dockerin domains mediate assembly of enzymes onto complementary cohesin motifs interspersed within non-catalytic protein scaffolds1. The versatile protein assembly mechanism conferred by the bacterial cohesin-dockerin interaction is now a standard design principle for synthetic protein-scale pathways2,3. For decades, analogous structures have been reported in the early branching anaerobic fungi, which are known to assemble by sequence divergent non-catalytic dockerin domains (NCDD)4. However, the enzyme components, modular assembly mechanism, and functional role of fungal cellulosomes remain unknown5,6. Here, we describe the comprehensive set of proteins critical to fungal cellulosome assembly, including novel, conserved scaffolding proteins unique to the Neocallimastigomycota. High quality genomes of the anaerobic fungi Anaeromyces robustus, Neocallimastix californiae and Piromyces finnis were assembled with long-read, single molecule technology to overcome their repeat-richness and extremely low GC content. Genomic analysis coupled with proteomic validation revealed an average 320 NCDD-containing proteins per fungal strain that were overwhelmingly carbohydrate active enzymes (CAZymes), with 95 large fungal scaffoldins identified across 4 genera that contain a conserved amino acid sequence repeat that binds to NCDDs. Fungal dockerin and scaffoldin domains have no similarity to their bacterial counterparts, yet several catalytic domains originated via horizontal gene transfer with gut bacteria. Though many catalytic domains are shared with bacteria, the biocatalytic activity of anaerobic fungi is expanded by the inclusion of GH3, GH6, and GH45 enzymes in the enzyme complexes. Collectively, these findings suggest that the fungal cellulosome is an evolutionarily

  4. Reaction of fungal products with amebocyte lysates of the Japanese horseshoe crab, Tachypleus tridentatus.

    Science.gov (United States)

    Hodes, D S; Heon, D; Hass, A; Hyatt, A C; Hodes, H L

    1987-09-01

    A commercially available endotoxin assay (CS-TAL) employing a chromogenic peptide and an amebocyte lysate from the Japanese horseshoe crab, Tachypleus tridentatus, gave a positive result with aqueous extracts of all 15 strains of Candida albicans and 1 strain each of Candida tropicalis, Cryptococcus neoformans, and a Mucor species that we tested. Purified glucans prepared from the Candida strains gave the same results. Reconstruction experiments showed that the positive results were not due to contaminating endotoxin. By contrast, assays employing amebocyte lysates of the American horseshoe crab, Limulus polyphemus, were inconsistent. Japanese workers have presented evidence that glucans activate the Tachypleus amebocyte lysate system by acting on an enzyme different from that on which endotoxin acts. Using a Tachypleus lysate preparation (Endospecy; Seikagaku Kogyo, Tokyo, Japan) from which this enzyme was excluded, we demonstrated a 5- to 10-fold drop in reactivity to the aqueous Candida extracts and glucans, whereas reactivity to endotoxin was unchanged. Normal human plasma was shown to decrease the effect of fungal extracts on CS-TAL. This inhibition was completely removed by heating the plasma. Our results suggest that Tachypleus systems may be of use clinically in distinguishing bacterial from fungal infections.

  5. Remediation of a contaminated soil by Ni+2 after application of biochar prepared from de-inking paper sludge: Influence on enzyme activities

    Science.gov (United States)

    Gascó, G.; Paz-Ferreiro, J.; Araujo, F.; Guerrero, F.; Méndez, A.

    2012-04-01

    In recent years, an increasing proportion of recycled fibres are used in paper industries due to their important environmental and economical benefits. A ton of pulp produced from recycled paper requires 60% less energy to manufacture than a ton of bleached virgin kraft pulp [1]. However, removing the ink, clay, coatings and contaminants from waste paper in order to produce recycled paper creates large amounts of de-inking paper sludge (DPS). Nowadays, more than 200000 t of DPS were produced in Spain. DPS can be used as amendment due to their high organic matter [2] but the high C/N ratio and the heavy metal content can limit its use. For this reason, the preparation of biochar obtained from pyrolysis process for water remediation [3] and soil contaminated by heavy metal can be an valorisation alternative. The main objective of this work is to study the influence of the biochar application prepared from de-inking sewage sludge in the soil enzyme activities of a contaminated soil by Ni+2 at two different concentrations. For this reason, an incubation experiment was performed and several enzymatic activities (dehydrogenase, b-glucosidase, phosphomoesterase and arylsulphatase) were monitored. The study was completed studying the influence of the biochar application in plant-available metals from soil. [1] Thompson C.G. 1992. Recycled Papers. The Essential Guide, MIT Press, Cambridge. [2] Barriga S., Méndez A., Cámara J., Guerrero F., Gascó G. 2010. Agricultural valorisation of de-inking paper sludge as organic amendment in different soils: Thermal study. Journal of Thermal Analysis and Calorimetry 99: 981-986 [3] Méndez A., Barriga S., Fidalgo J.M., Gascó G. 2009. Adsorbent materials from paper industry waste materials and their use in Cu(II) removal from water. Journal of Hazardous Materials 165: 736-743.

  6. Metal-organic coordination-enabled layer-by-layer self-assembly to prepare hybrid microcapsules for efficient enzyme immobilization.

    Science.gov (United States)

    Wang, Xiaoli; Jiang, Zhongyi; Shi, Jiafu; Liang, Yanpeng; Zhang, Chunhong; Wu, Hong

    2012-07-25

    A novel layer-by-layer self-assembly approach enabled by metal-organic coordination was developed to prepare polymer-inorganic hybrid microcapsules. Alginate was first activated via N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxy succinimide (NHS) coupling chemistry, and subsequently reacted with dopamine. Afterward, the dopamine modified alginate (Alg-DA) and titanium(IV) bis(ammonium lactato) dihydroxide (Ti(IV)) were alternatively deposited onto CaCO3 templates. The coordination reaction between the catechol groups of Alg-DA and the Ti(IV) allowed the alternative assembly to form a series of multilayers. After removing the templates, the alginate-titanium hybrid microcapsules were obtained. The high mechanical stability of hybrid microcapsules was demonstrated by osmotic pressure experiment. Furthermore, the hybrid microcapsules displayed superior thermal stability due to Ti(IV) coordination. Catalase (CAT) was used as model enzyme, either encapsulated inside or covalently attached on the surface of the resultant microcapsules. No CAT leakage from the microcapsules was detected after incubation for 48 h. The encapsulated CAT, with a loading capacity of 450-500 mg g(-1) microcapsules, exhibited desirable long-term storage stability, whereas the covalently attached CAT, with a loading capacity of 100-150 mg g(-1) microcapsules, showed desirable operational stability.

  7. Fungal Genomics Program

    Energy Technology Data Exchange (ETDEWEB)

    Grigoriev, Igor

    2012-03-12

    The JGI Fungal Genomics Program aims to scale up sequencing and analysis of fungal genomes to explore the diversity of fungi important for energy and the environment, and to promote functional studies on a system level. Combining new sequencing technologies and comparative genomics tools, JGI is now leading the world in fungal genome sequencing and analysis. Over 120 sequenced fungal genomes with analytical tools are available via MycoCosm (www.jgi.doe.gov/fungi), a web-portal for fungal biologists. Our model of interacting with user communities, unique among other sequencing centers, helps organize these communities, improves genome annotation and analysis work, and facilitates new larger-scale genomic projects. This resulted in 20 high-profile papers published in 2011 alone and contributing to the Genomics Encyclopedia of Fungi, which targets fungi related to plant health (symbionts, pathogens, and biocontrol agents) and biorefinery processes (cellulose degradation, sugar fermentation, industrial hosts). Our next grand challenges include larger scale exploration of fungal diversity (1000 fungal genomes), developing molecular tools for DOE-relevant model organisms, and analysis of complex systems and metagenomes.

  8. Fungal DNA barcoding.

    Science.gov (United States)

    Xu, Jianping

    2016-11-01

    Fungi are ubiquitous in both natural and human-made environments. They play important roles in the health of plants, animals, and humans, and in broad ecosystem functions. Thus, having an efficient species-level identification system could significantly enhance our ability to treat fungal diseases and to monitor the spatial and temporal patterns of fungal distributions and migrations. DNA barcoding is a potent approach for rapid identification of fungal specimens, generating novel species hypothesis, and guiding biodiversity and ecological studies. In this mini-review, I briefly summarize (i) the history of DNA sequence-based fungal identification; (ii) the emergence of the ITS region as the consensus primary fungal barcode; (iii) the use of the ITS barcodes to address a variety of issues on fungal diversity from local to global scales, including generating a large number of species hypothesis; and (iv) the problems with the ITS barcode region and the approaches to overcome these problems. Similar to DNA barcoding research on plants and animals, significant progress has been achieved over the last few years in terms of both the questions being addressed and the foundations being laid for future research endeavors. However, significant challenges remain. I suggest three broad areas of research to enhance the usefulness of fungal DNA barcoding to meet the current and future challenges: (i) develop a common set of primers and technologies that allow the amplification and sequencing of all fungi at both the primary and secondary barcode loci; (ii) compile a centralized reference database that includes all recognized fungal species as well as species hypothesis, and allows regular updates from the research community; and (iii) establish a consensus set of new species recognition criteria based on barcode DNA sequences that can be applied across the fungal kingdom.

  9. Fungal symbiosis unearthed

    Science.gov (United States)

    Daniel Cullen

    2008-01-01

    Associations between plant roots and fungi are a feature of many terrestrial ecosystems. The genome sequence of a prominent fungal partner opens new avenues for studying such mycorrhizal interactions....

  10. Fungal Skin Infections

    Science.gov (United States)

    ... Abbreviations Weights & Measures ENGLISH View Professional English Deutsch Japanese Espaniol Find information on medical topics, symptoms, drugs, ... touching the infected area. Diagnosis Skin scrapings or cultures Doctors may suspect a fungal infection when they ...

  11. JGI Fungal Genomics Program

    Energy Technology Data Exchange (ETDEWEB)

    Grigoriev, Igor V.

    2011-03-14

    Genomes of energy and environment fungi are in focus of the Fungal Genomic Program at the US Department of Energy Joint Genome Institute (JGI). Its key project, the Genomics Encyclopedia of Fungi, targets fungi related to plant health (symbionts, pathogens, and biocontrol agents) and biorefinery processes (cellulose degradation, sugar fermentation, industrial hosts), and explores fungal diversity by means of genome sequencing and analysis. Over 50 fungal genomes have been sequenced by JGI to date and released through MycoCosm (www.jgi.doe.gov/fungi), a fungal web-portal, which integrates sequence and functional data with genome analysis tools for user community. Sequence analysis supported by functional genomics leads to developing parts list for complex systems ranging from ecosystems of biofuel crops to biorefineries. Recent examples of such 'parts' suggested by comparative genomics and functional analysis in these areas are presented here

  12. [Allergic fungal sinusitis].

    Science.gov (United States)

    Gras, J R; Lafarga, J; Ronda, J M; Trigueros, M; Sancho, M; Aracil, A

    2000-10-01

    Allergic fungal sinusitis is a recently described clinical entity that has gained increased attention as a cause of chronic sinusitis. Consist in a benign noninvasive sinus disease related to a hypersensitivity reaction to fungal antigens. It should be suspected in any atopic patient with refractory nasal polyps. Computed tomography (CT) findings are characteristics, but not diagnostic. Diagnosis requires show allergic mucin in the histopathologic examination and hiphae in special fungal stains. The suitable treatment includes the allergic mucin removal and sinus aeration accomplished endoscopically, perioperative systemic steroids and immunotherapy with fungal antigens. We present a case of this kind of chronic sinusitis describing the characteristic histopathologic and radiologic findings, the pathogenic theories and recent advances in immunotherapy.

  13. Fungal toenail infections

    OpenAIRE

    Ferrari, Jill

    2014-01-01

    Fungal toenail infection (onychomycosis) is characterised as infection of part or all of the toenail unit, which includes the nail plate, the nail bed, and the nail matrix. Over time, the infection causes discoloration and distortion of part or all of the nail unit. Fungal infections are reported to cause 23% of foot diseases and 50% of nail conditions in people seen by dermatologists, but are less common in the general population, affecting 3% to 12% of people.Infection can cause discomfo...

  14. Fungal toenail infections

    OpenAIRE

    Ferrari, Jill

    2011-01-01

    Fungal toenail infection (onychomycosis) is characterised as infection of part or all of the toenail unit, which includes the nail plate, the nail bed, and the nail matrix. Over time, the infection causes discoloration and distortion of part or all of the nail unit. Fungal infections are reported to cause 23% of foot diseases and 50% of nail conditions in people seen by dermatologists, but are less common in the general population, affecting 3% to 5% of people.Infection can cause discomfor...

  15. Inositol Polyphosphate Kinases, Fungal Virulence and Drug Discovery.

    Science.gov (United States)

    Li, Cecilia; Lev, Sophie; Saiardi, Adolfo; Desmarini, Desmarini; Sorrell, Tania C; Djordjevic, Julianne T

    2016-09-06

    Opportunistic fungi are a major cause of morbidity and mortality world-wide, particularly in immunocompromised individuals. Developing new treatments to combat invasive fungal disease is challenging given that fungal and mammalian host cells are eukaryotic, with similar organization and physiology. Even therapies targeting unique fungal cell features have limitations and drug resistance is emerging. New approaches to the development of antifungal drugs are therefore needed urgently. Cryptococcus neoformans , the commonest cause of fungal meningitis worldwide, is an accepted model for studying fungal pathogenicity and driving drug discovery. We recently characterized a phospholipase C (Plc1)-dependent pathway in C. neoformans comprising of sequentially-acting inositol polyphosphate kinases (IPK), which are involved in synthesizing inositol polyphosphates (IP). We also showed that the pathway is essential for fungal cellular function and pathogenicity. The IP products of the pathway are structurally diverse, each consisting of an inositol ring, with phosphate (P) and pyrophosphate (PP) groups covalently attached at different positions. This review focuses on (1) the characterization of the Plc1/IPK pathway in C. neoformans ; (2) the identification of PP-IP₅ (IP₇) as the most crucial IP species for fungal fitness and virulence in a mouse model of fungal infection; and (3) why IPK enzymes represent suitable candidates for drug development.

  16. Inositol Polyphosphate Kinases, Fungal Virulence and Drug Discovery

    Directory of Open Access Journals (Sweden)

    Cecilia Li

    2016-09-01

    Full Text Available Opportunistic fungi are a major cause of morbidity and mortality world-wide, particularly in immunocompromised individuals. Developing new treatments to combat invasive fungal disease is challenging given that fungal and mammalian host cells are eukaryotic, with similar organization and physiology. Even therapies targeting unique fungal cell features have limitations and drug resistance is emerging. New approaches to the development of antifungal drugs are therefore needed urgently. Cryptococcus neoformans, the commonest cause of fungal meningitis worldwide, is an accepted model for studying fungal pathogenicity and driving drug discovery. We recently characterized a phospholipase C (Plc1-dependent pathway in C. neoformans comprising of sequentially-acting inositol polyphosphate kinases (IPK, which are involved in synthesizing inositol polyphosphates (IP. We also showed that the pathway is essential for fungal cellular function and pathogenicity. The IP products of the pathway are structurally diverse, each consisting of an inositol ring, with phosphate (P and pyrophosphate (PP groups covalently attached at different positions. This review focuses on (1 the characterization of the Plc1/IPK pathway in C. neoformans; (2 the identification of PP-IP5 (IP7 as the most crucial IP species for fungal fitness and virulence in a mouse model of fungal infection; and (3 why IPK enzymes represent suitable candidates for drug development.

  17. Magnetically responsive enzyme powders

    International Nuclear Information System (INIS)

    Pospiskova, Kristyna; Safarik, Ivo

    2015-01-01

    Powdered enzymes were transformed into their insoluble magnetic derivatives retaining their catalytic activity. Enzyme powders (e.g., trypsin and lipase) were suspended in various liquid media not allowing their solubilization (e.g., saturated ammonium sulfate and highly concentrated polyethylene glycol solutions, ethanol, methanol, 2-propanol) and subsequently cross-linked with glutaraldehyde. Magnetic modification was successfully performed at low temperature in a freezer (−20 °C) using magnetic iron oxides nano- and microparticles prepared by microwave-assisted synthesis from ferrous sulfate. Magnetized cross-linked enzyme powders were stable at least for two months in water suspension without leakage of fixed magnetic particles. Operational stability of magnetically responsive enzymes during eight repeated reaction cycles was generally without loss of enzyme activity. Separation of magnetically modified cross-linked powdered enzymes from reaction mixtures was significantly simplified due to their magnetic properties. - Highlights: • Cross-linked enzyme powders were prepared in various liquid media. • Insoluble enzymes were magnetized using iron oxides particles. • Magnetic iron oxides particles were prepared by microwave-assisted synthesis. • Magnetic modification was performed under low (freezing) temperature. • Cross-linked powdered trypsin and lipase can be used repeatedly for reaction

  18. Pectic enzymes

    NARCIS (Netherlands)

    Benen, J.A.E.; Voragen, A.G.J.; Visser, J.

    2003-01-01

    The pectic enzymes comprise a diverse group of enzymes. They consist of main-chain depolymerases and esterases active on methyl- and acetylesters of galacturonosyl uronic acid residues. The depolymerizing enzymes comprise hydrolases as wel as lyases

  19. Cross-linked enzyme aggregates (CLEAs) of halohydrin dehalogenase from Agrobacterium radiobacter AD1: Preparation, characterization and application as a biocatalyst.

    Science.gov (United States)

    Liao, Qian; Du, Xuan; Jiang, Wei; Tong, Yapei; Zhao, Zhipeng; Fang, Ruiqin; Feng, Juan; Tang, Lixia

    2017-12-19

    Halohydrin dehalogenase from Agrobacterium radiobacter AD1 (HheC) shows great potential to produce valuable optically pure epoxides and β-substituted alcohols. However, this enzyme has been reported to be very sensitive and less stable under oxidative conditions. Enzyme immobilization represents a powerful means to overcome this limitation and provides the enzyme characteristics of a biocatalyst. In this study, the crude extract of HheC was directly subjected to enzyme immobilization using a carrier-free cross-linked enzyme aggregates (CLEAs) method. The results showed that under the optimized conditions, the obtained HheC-CLEAs retained more than 90% activity of the free enzyme; preserved more than 50% of their original activity after storage at 4 °C for 2 months, even in the absence of a reducing agent; displayed a strong tolerance to organic solvents with fully active after incubation in the presence of 50% cyclohexane and n-hexane for 5 hours; the presence of organic solvents could minimize the negative effect of enzyme immobilization on the enzntioselectivity of HheC. Most importantly, HheC-CLEAs maintained more than 70% activity after 10 reusability cycles. The utility of HheC-CLEAs as a valuable biocatalyst was exhibited by the kinetic resolution of azide-mediated ring-opening reaction of rac-1,2-epoxy-2-methylbutane. These results indicated that HheC-CLEAs overcame some disadvantages of free enzymes to become biocatalysts. Together with further engineering of the enzyme, HheC-CLEAs could become a promising biocatalyst for the synthesis of valuable chiral compounds. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Preparation and characterization of domoic acid-protein conjugates using small amount of toxin in a reversed micellar medium: application in a competitive enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Branaa, P; Naar, J; Chinain, M; Pauillac, S

    1999-01-01

    With the aim of producing novel antibodies to domoic acid (DA), an original, rapid, and simple procedure for preparing minute amount of hapten-protein conjugates was developed. The amide-bond-generating mixed anhydride method of Erlanger was performed using 0.32-0.64 micromol of DA in a reversed micellar medium allowing strong carrier haptenization as determined by spectrophotometric measurement. Bovine serum albumin (BSA) and ovalbumin (OVA) conjugates were, respectively, used for immunization of BALB/c mice and antibody screening by enzyme-linked immunosorbent assay (ELISA). Specific polyclonal antibodies were produced upon multiple injections of (DA)(17)-BSA conjugate administered by three different routes: (i) intraperitoneal (i.p.), (ii) intraperitoneal + subcutaneous (i.p. + s.c.), (iii) footpad (f.p.). The i.p. route induced antisera of higher titer (1:350000) than did the other protocols (approximately 1:72900) and was selected throughout further experiments. Using a competitive ELISA format with a peroxidase immunoconjugate and a chromogenic substrate, no significant cross-reactivity was observed with glutamic acid, aspartic acid and kainic acid (KA), a structural analogue of DA. The sensitivity of this assay could be enhanced by 1 order of magnitude by using a beta-galactosidase immunoconjugate with a fluorogenic substrate while preserving DA specificity. The calculated dissociation constant (K(D)) for the interaction of the antibodies with free DA was 5 x 10(-)(7) M (chromogenic assay) and 5 x 10(-)(8) M (fluorogenic assay). Using the optimized assay the limit of detection (LOD) and the limit of quantitation (LOQ) in the ELISA buffer were 1.4 and 3 ng/mL, respectively. Moreover this assay was found applicable for measuring DA levels in spiked mussel extracts pre-cleaned through a solid-phase extraction column, as a very good correlation (r(2) = 0.96) was observed between the actual amounts of DA added and amounts detected by ELISA. Thus, accurate

  1. Fungal polygalacturonase activity reflects susceptibility of carnation cultivars to Fusarium wilt

    NARCIS (Netherlands)

    Baayen, R.P.; Schoffelmeer, E.A.M.; Toet, S.; Elgersma, D.M.

    1997-01-01

    Carnation cultivars with different levels of partial resistance were inoculated with race 2 of Fusarium oxysporum f.sp. dianthi and monitored for accumulation of host phytoalexins, fungal escape from compartmentalization, production of fungal pectin-degrading enzymes and development of external

  2. Fungal toenail infections

    OpenAIRE

    Ferrari, Jill

    2008-01-01

    Fungal toenail infection (onychomycosis) is characterised as infection of part or all of the toenail unit, which includes the nail plate, the nail bed, and the nail matrix. Over time, the infection causes discoloration and distortion of part or all of the nail unit. Fungal infections are reported to cause 23% of foot diseases and 50% of nail conditions in people seen by dermatologists, but are less common in the general population, affecting 3-5% of people.Infection can cause discomfort in...

  3. Magnetically responsive enzyme powders

    Science.gov (United States)

    Pospiskova, Kristyna; Safarik, Ivo

    2015-04-01

    Powdered enzymes were transformed into their insoluble magnetic derivatives retaining their catalytic activity. Enzyme powders (e.g., trypsin and lipase) were suspended in various liquid media not allowing their solubilization (e.g., saturated ammonium sulfate and highly concentrated polyethylene glycol solutions, ethanol, methanol, 2-propanol) and subsequently cross-linked with glutaraldehyde. Magnetic modification was successfully performed at low temperature in a freezer (-20 °C) using magnetic iron oxides nano- and microparticles prepared by microwave-assisted synthesis from ferrous sulfate. Magnetized cross-linked enzyme powders were stable at least for two months in water suspension without leakage of fixed magnetic particles. Operational stability of magnetically responsive enzymes during eight repeated reaction cycles was generally without loss of enzyme activity. Separation of magnetically modified cross-linked powdered enzymes from reaction mixtures was significantly simplified due to their magnetic properties.

  4. Synthetic biology of fungal natural products

    Science.gov (United States)

    Mattern, Derek J.; Valiante, Vito; Unkles, Shiela E.; Brakhage, Axel A.

    2015-01-01

    Synthetic biology is an ever-expanding field in science, also encompassing the research area of fungal natural product (NP) discovery and production. Until now, different aspects of synthetic biology have been covered in fungal NP studies from the manipulation of different regulatory elements and heterologous expression of biosynthetic pathways to the engineering of different multidomain biosynthetic enzymes such as polyketide synthases or non-ribosomal peptide synthetases. The following review will cover some of the exemplary studies of synthetic biology in filamentous fungi showing the capacity of these eukaryotes to be used as model organisms in the field. From the vast array of different NPs produced to the ease for genetic manipulation, filamentous fungi have proven to be an invaluable source for the further development of synthetic biology tools. PMID:26284053

  5. Optimization of Culture Conditions for Some Identified Fungal Species and Stability Profile of α-Galactosidase Produced

    Directory of Open Access Journals (Sweden)

    A. S. Chauhan

    2013-01-01

    Full Text Available Microbial α-galactosidase preparations have implications in medicine and in the modification of various agricultural products as well. In this paper, four isolated fungal strains such as AL-3, WF-3, WP-4 and CL-4 from rhizospheric soil identified as Penicillium glabrum (AL-3, Trichoderma evansii (WF-3, Lasiodiplodia theobromae (WP-4 and Penicillium flavus (CL-4 based on their morphology and microscopic examinations, are screened for their potential towards α-galactosidases production. The culture conditions have been optimized and supplemented with specific carbon substrates (1%, w/v by using galactose-containing polysaccharides like guar gum (GG, soya casein (SC and wheat straw (WS. All strains significantly released galactose from GG, showing maximum production of enzyme at 7th day of incubation in rotary shaker (120 rpm that is 190.3, 174.5, 93.9 and 28.8 U/mL, respectively, followed by SC and WS. The enzyme activity was stable up to 7days at −20°C, then after it declines. This investigation reveals that AL-3 show optimum enzyme activity in guar gum media, whereas WF-3 exhibited greater enzyme stability. Results indicated that the secretion of proteins, enzyme and the stability of enzyme activity varied not only from one strain to another but also differed in their preferences of utilization of different substrates.

  6. Mycelial growth interactions and mannan-degrading enzyme ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-18

    May 18, 2009 ... 1Department of Land Management, Faculty of Agriculture, Universiti Putra Malaysia 43400-UPM, Serdang, Selangor,. Malaysia. ... For mannan- degrading enzyme production, single and co-cultures of these fungi were carried out under submerged ... Fungal species and combinations. The 5 fungal strains ...

  7. Immunoregulation in Fungal Diseases

    Directory of Open Access Journals (Sweden)

    Jonathan A. Roussey

    2016-12-01

    Full Text Available This review addresses specific regulatory mechanisms involved in the host immune response to fungal organisms. We focus on key cells and regulatory pathways involved in these responses, including a brief overview of their broader function preceding a discussion of their specific relevance to fungal disease. Important cell types discussed include dendritic cells and regulatory T cells, with a focus on specific studies relating to their effects on immune responses to fungi. We highlight the interleukin-10, programmed cell death 1, and cytotoxic T lymphocyte-associated protein 4 signaling pathways and emphasize interrelationships between these pathways and the regulatory functions of dendritic cells and regulatory T cells. Throughout our discussion, we identify selected studies best illustrating the role of these cells and pathways in response to specific fungal pathogens to provide a contextual understanding of the tightly-controlled network of regulatory mechanisms critical to determining the outcome of exposure to fungal pathogens. Lastly, we discuss two unique phenomena relating to immunoregulation, protective tolerance and immune reactivation inflammatory syndrome. These two clinically-relevant conditions provide perspective as to the range of immunoregulatory mechanisms active in response to fungi.

  8. Allergic Fungal Airway Disease.

    Science.gov (United States)

    Rick, E M; Woolnough, K; Pashley, C H; Wardlaw, A J

    Fungi are ubiquitous and form their own kingdom. Up to 80 genera of fungi have been linked to type I allergic disease, and yet, commercial reagents to test for sensitization are available for relatively few species. In terms of asthma, it is important to distinguish between species unable to grow at body temperature and those that can (thermotolerant) and thereby have the potential to colonize the respiratory tract. The former, which include the commonly studied Alternaria and Cladosporium genera, can act as aeroallergens whose clinical effects are predictably related to exposure levels. In contrast, thermotolerant species, which include fungi from the Candida, Aspergillus, and Penicillium genera, can cause a persistent allergenic stimulus independent of their airborne concentrations. Moreover, their ability to germinate in the airways provides a more diverse allergenic stimulus, and may result in noninvasive infection, which enhances inflammation. The close association between IgE sensitization to thermotolerant filamentous fungi and fixed airflow obstruction, bronchiectasis, and lung fibrosis suggests a much more tissue-damaging process than that seen with aeroallergens. This review provides an overview of fungal allergens and the patterns of clinical disease associated with exposure. It clarifies the various terminologies associated with fungal allergy in asthma and makes the case for a new term (allergic fungal airway disease) to include all people with asthma at risk of developing lung damage as a result of their fungal allergy. Lastly, it discusses the management of fungirelated asthma.

  9. Thai marine fungal diversity

    OpenAIRE

    Rattaket Choeyklin; Souwalak Phongpaichit; Ittichai Chatmala; Jariya Sakayaroj; Apiradee Pilantanapak; E.B. Gareth Jones

    2006-01-01

    The marine fungal diversity of Thailand was investigated and 116 Ascomycota, 3 Basidiomycota, 28 anamorphic fungi, 7 Stramenopiles recorded, with 30 tentatively identified. These species have primarily been collected from driftwood and attached decayed wood of mangrove trees. The holotype number of 15 taxa is from Thailand and 33 are new records from the country.

  10. Thai marine fungal diversity

    Directory of Open Access Journals (Sweden)

    Rattaket Choeyklin

    2006-07-01

    Full Text Available The marine fungal diversity of Thailand was investigated and 116 Ascomycota, 3 Basidiomycota, 28 anamorphic fungi, 7 Stramenopiles recorded, with 30 tentatively identified. These species have primarily been collected from driftwood and attached decayed wood of mangrove trees. The holotype number of 15 taxa is from Thailand and 33 are new records from the country.

  11. The Fungal Kingdom

    NARCIS (Netherlands)

    Heitman, Joseph; Howlett, B.J.; Crous, P.W.; Stukenbrock, E.H.; James, T.Y.; Gow, N.A.R.

    2017-01-01

    Fungi research and knowledge grew rapidly following recent advances in genetics and genomics. This book synthesizes new knowledge with existing information to stimulate new scientific questions and propel fungal scientists on to the next stages of research. This book is a comprehensive guide on

  12. Fungal Wound Infection

    Centers for Disease Control (CDC) Podcasts

    2016-01-28

    Dr. David Tribble, acting director of the infectious disease clinical research program at Uniformed Services University of the Health Sciences, discusses fungal wound infections after combat trauma.  Created: 1/28/2016 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 1/28/2016.

  13. Autochthonous fungal strains with high ligninolytic activities from ...

    African Journals Online (AJOL)

    These fungal strains were first screened for lignin-modifying enzymes on solid media containing Poly R-478 or ABTS. Of the 315 tested strains, 49 exhibited significant ABTS-oxidation activity, expressed within the first week of incubation and only 18 strains decolourised the Poly R-478. These positive strains were further ...

  14. Generation of transgenic mice producing fungal xylanase in the ...

    African Journals Online (AJOL)

    To our knowledge, this is the first demonstration of the production of fungal xylanase in the saliva of simple-stomached animals. Results from the present study encourage further investigation of employing transgenic technology to enhance the digestive capability of monogastric agriculture animals by introducing enzyme ...

  15. Whole-cell fungal transformation of precursors into dyes

    Science.gov (United States)

    2010-01-01

    Background Chemical methods of producing dyes involve extreme temperatures and unsafe toxic compounds. Application of oxidizing enzymes obtained from fungal species, for example laccase, is an alternative to chemical synthesis of dyes. Laccase can be replaced by fungal biomass acting as a whole-cell biocatalyst with properties comparable to the isolated form of the enzyme. The application of the whole-cell system simplifies the transformation process and reduces the time required for its completion. In the present work, four fungal strains with a well-known ability to produce laccase were tested for oxidation of 17 phenolic and non-phenolic precursors into stable and non-toxic dyes. Results An agar-plate screening test of the organic precursors was carried out using four fungal strains: Trametes versicolor, Fomes fomentarius, Abortiporus biennis, and Cerrena unicolor. Out of 17 precursors, nine were transformed into coloured substances in the presence of actively growing fungal mycelium. The immobilized fungal biomass catalyzed the transformation of 1 mM benzene and naphthalene derivatives in liquid cultures yielding stable and non-toxic products with good dyeing properties. The type of fungal strain had a large influence on the absorbance of the coloured products obtained after 48-hour transformation of the selected precursors, and the most effective was Fomes fomentarius (FF25). Whole-cell transformation of AHBS (3-amino-4-hydroxybenzenesulfonic acid) into a phenoxazinone dye was carried out in four different systems: in aqueous media comprising low amounts of carbon and nitrogen source, in buffer, and in distilled water. Conclusions This study demonstrated the ability of four fungal strains belonging to the ecological type of white rot fungi to transform precursors into dyes. This paper highlights the potential of fungal biomass for replacing isolated enzymes as a cheaper industrial-grade biocatalyst for the synthesis of dyes and other commercially important

  16. Whole-cell fungal transformation of precursors into dyes

    Directory of Open Access Journals (Sweden)

    Jarosz-Wilkołazka Anna

    2010-07-01

    Full Text Available Abstract Background Chemical methods of producing dyes involve extreme temperatures and unsafe toxic compounds. Application of oxidizing enzymes obtained from fungal species, for example laccase, is an alternative to chemical synthesis of dyes. Laccase can be replaced by fungal biomass acting as a whole-cell biocatalyst with properties comparable to the isolated form of the enzyme. The application of the whole-cell system simplifies the transformation process and reduces the time required for its completion. In the present work, four fungal strains with a well-known ability to produce laccase were tested for oxidation of 17 phenolic and non-phenolic precursors into stable and non-toxic dyes. Results An agar-plate screening test of the organic precursors was carried out using four fungal strains: Trametes versicolor, Fomes fomentarius, Abortiporus biennis, and Cerrena unicolor. Out of 17 precursors, nine were transformed into coloured substances in the presence of actively growing fungal mycelium. The immobilized fungal biomass catalyzed the transformation of 1 mM benzene and naphthalene derivatives in liquid cultures yielding stable and non-toxic products with good dyeing properties. The type of fungal strain had a large influence on the absorbance of the coloured products obtained after 48-hour transformation of the selected precursors, and the most effective was Fomes fomentarius (FF25. Whole-cell transformation of AHBS (3-amino-4-hydroxybenzenesulfonic acid into a phenoxazinone dye was carried out in four different systems: in aqueous media comprising low amounts of carbon and nitrogen source, in buffer, and in distilled water. Conclusions This study demonstrated the ability of four fungal strains belonging to the ecological type of white rot fungi to transform precursors into dyes. This paper highlights the potential of fungal biomass for replacing isolated enzymes as a cheaper industrial-grade biocatalyst for the synthesis of dyes and other

  17. [Clinically documented fungal infections].

    Science.gov (United States)

    Kakeya, Hiroshi; Kohno, Shigeru

    2008-12-01

    Proven fungal infections are diagnosed by histological/microbiological evidence of fungi at the site of infection and positive blood culture (fungemia). However, invasive diagnosing examinations are not always applied for all of immunocompromised patients. Clinically documented invasive fungal infections are diagnosed by typical radiological findings such as halo sign on chest CT plus positive serological/molecular evidence of fungi. Serological tests of Aspergillus galactomannan antigen and beta-glucan for aspergillosis and cryptococcal glucuronoxylomannan antigen for cryptococcosis are useful. Hence, none of reliable serological tests for zygomycosis are available so far. In this article, risk factors, sign and symptoms, and diagnostic methods for clinically documented cases of invasive aspergillosis, pulmonary cryptococcosis, and zygomycosis with diabates, are reviewed.

  18. Treatment of superficial fungal infections – recommendations of experts of Mycological Section of Polish Dermatological Society

    OpenAIRE

    Romuald Maleszka; Zygmunt Adamski; Jacek Szepietowski; Eugeniusz Baran

    2015-01-01

    Fungal infections are very commonly occurring diseases in everyday dermatological practice, and knowledge of the wide spectrum of antifungal preparations available nowadays is helpful in almost all medical disciplines. The treatment of fungal infections should be preceded with a proper diagnosis based on the patient’s history, precise clinical examination and diagnostic mycological procedures. A correct diagnosis of fungal infection is possible by its confirmation with proper investigations c...

  19. Metals in fungal virulence.

    Science.gov (United States)

    Gerwien, Franziska; Skrahina, Volha; Kasper, Lydia; Hube, Bernhard; Brunke, Sascha

    2018-01-01

    Metals are essential for life, and they play a central role in the struggle between infecting microbes and their hosts. In fact, an important aspect of microbial pathogenesis is the 'nutritional immunity', in which metals are actively restricted (or, in an extended definition of the term, locally enriched) by the host to hinder microbial growth and virulence. Consequently, fungi have evolved often complex regulatory networks, uptake and detoxification systems for essential metals such as iron, zinc, copper, nickel and manganese. These systems often differ fundamentally from their bacterial counterparts, but even within the fungal pathogens we can find common and unique solutions to maintain metal homeostasis. Thus, we here compare the common and species-specific mechanisms used for different metals among different fungal species-focusing on important human pathogens such as Candida albicans, Aspergillus fumigatus or Cryptococcus neoformans, but also looking at model fungi such as Saccharomyces cerevisiae or A. nidulans as well-studied examples for the underlying principles. These direct comparisons of our current knowledge reveal that we have a good understanding how model fungal pathogens take up iron or zinc, but that much is still to learn about other metals and specific adaptations of individual species-not the least to exploit this knowledge for new antifungal strategies. © FEMS 2017.

  20. Metarhizium anisopliae: influence of pH on enzyme activity and control of Rhipicephalus microplus ticks

    Directory of Open Access Journals (Sweden)

    Allan Felipe Marciano

    2015-12-01

    Full Text Available ABSTRACT. Marciano A.F., Coutinho-Rodrigues C.J.B., Perinotto W.M.S., Camargo M.G., Gôlo P.S., Sá F.A., Quinelato S., Freitas M.C., Angelo I.C., Nogueira M.R.S. & Bittencourt V.R.E.P. [Metarhizium anisopliae: influence of pH on enzyme activity and control of Rhipicephalus microplus ticks.] Metarhizium anisopliae: influência do pH na atividade enzimática e no controle de Rhipicephalus microplus. Revista Brasileira de Medicina Veterinária, 37(Supl.1:85-90, 2015. Departamento de Parasitologia Animal, Universidade Federal Rural do Rio de Janeiro, BR 465, Km 47, Seropédica, RJ 23897-970, Brasil. E-mail: vaniabit@ufrrj.br Rhipicephalus microplus ticks are one of the major agents causing substantial losses to livestock worldwide. In the search for alternative control strategies, both in vitro and in vivo use of the arthropodpathogenic fungus Metarhizium anisopliae has shown promising results against this ectoparasite. During host colonization, protease production by M. anisopliae is considered one important virulence factor once it is directly related to the active penetration process carried by the fungus on the full host cuticle. Nevertheless, limitations as environmental pH may modulate the proteases production and/or activity, as well as, the fungal virulence. The current study aimed evaluate the virulence and total protease activity of M. anisopliae CG 148 sensu lato (s.l.. Fungal aqueous suspensions or 5% mineral oil formulations were used in different pH ranges (5, 7, or 9. Suspensions and formulations were prepared using a pH meter and adjusted to 108 spores mL-1. In the bioassay, four groups were formed for each pH range: the aqueous fungal suspension, the oil-based fungal formulation and their respective controls (aqueous and oil-based, totaling 12 groups. Engorged females were immersed for 3 minutes and maintained under optimal conditions for evaluation of biological parameters. Total protease activity of the artificial medium (after

  1. Fertility, mortality, milk output, and body thermoregulation of growing Hy-Plus rabbits fed on diets supplemented with multi-enzymes preparation.

    Science.gov (United States)

    Gado, Hany M; Kholif, Ahmed E; Salem, Abdelfattah Z M; Elghandour, Mona M M; Olafadehan, Oluwarotimi A; Martinez, Maricela A; Al-Momani, Ahmed Q

    2016-10-01

    The aim of this study was to evaluate the fertility status, milk output, mortality, and body thermoregulation of rabbit does as affected by different levels of multi-enzyme extracts (EZ) in their diets. A total of 120 Hy-Plus rabbit does were divided into four comparable experimental groups (n = 30 does per group). Animals of each group were divided in six pens (five animals per pen), and each pen was used as an experimental unit. The first group was kept untreated and fed a commercial diet alone without enzyme extracts (EZ0), while the other groups were fed the same diet but supplemented with 1 (EZ1), 3 (EZ3), and 5 (EZ5) kg/ton of enzyme extracts, respectively. Feeding EZ additive increased (P < 0.05) conception and kindling rates, litter size and weight at birth, and litter size and bunny weight at weaning, with decreasing (P < 0.05) abortion rate. Moreover, total milk yield increased (P < 0.05) with increasing level of enzyme supplementation. Pre-weaning mortality decreased (P < 0.05) with EZ inclusion. Signs of vitality (rectal temperature, skin temperature, earlobe temperature, respiration rate, and pulse rate) were improved with EZ inclusion. For all results, 5 kg EZ/ton of feed was more effective than 1 and 3 kg EZ/ton feed. It can be concluded that supplementation of EZ in rabbit diet decreased mortality rate and enhanced fertility status and milk output.

  2. Enzymes in Fermented Fish.

    Science.gov (United States)

    Giyatmi; Irianto, H E

    Fermented fish products are very popular particularly in Southeast Asian countries. These products have unique characteristics, especially in terms of aroma, flavor, and texture developing during fermentation process. Proteolytic enzymes have a main role in hydrolyzing protein into simpler compounds. Fermentation process of fish relies both on naturally occurring enzymes (in the muscle or the intestinal tract) as well as bacteria. Fermented fish products processed using the whole fish show a different characteristic compared to those prepared from headed and gutted fish. Endogenous enzymes like trypsin, chymotrypsin, elastase, and aminopeptidase are the most involved in the fermentation process. Muscle tissue enzymes like cathepsins, peptidases, transaminases, amidases, amino acid decarboxylases, glutamic dehydrogenases, and related enzymes may also play a role in fish fermentation. Due to the decreased bacterial number during fermentation, contribution of microbial enzymes to proteolysis may be expected prior to salting of fish. Commercial enzymes are supplemented during processing for specific purposes, such as quality improvement and process acceleration. In the case of fish sauce, efforts to accelerate fermentation process and to improve product quality have been studied by addition of enzymes such as papain, bromelain, trypsin, pepsin, and chymotrypsin. © 2017 Elsevier Inc. All rights reserved.

  3. New insights into the formation of fungal aromatic polyketides.

    Science.gov (United States)

    Crawford, Jason M; Townsend, Craig A

    2010-12-01

    Fungal aromatic polyketides constitute a large family of bioactive natural products and are synthesized by the non-reducing group of iterative polyketide synthases (PKSs). Their diverse structures arise from selective enzymatic modifications of reactive, enzyme-bound poly-β-keto intermediates. How iterative PKSs control starter unit selection, polyketide chain initiation and elongation, intermediate folding and cyclization, selective redox or modification reactions during assembly, and product release are central mechanistic questions underlying iterative catalysis. This Review highlights recent insights into these questions, with a particular focus on the biosynthetic programming of fungal aromatic polyketides, and draws comparisons with the allied biosynthetic processes in bacteria.

  4. Phylogenomic relationships between amylolytic enzymes from 85 strains of fungi.

    Directory of Open Access Journals (Sweden)

    Wanping Chen

    Full Text Available Fungal amylolytic enzymes, including α-amylase, gluocoamylase and α-glucosidase, have been extensively exploited in diverse industrial applications such as high fructose syrup production, paper making, food processing and ethanol production. In this paper, amylolytic genes of 85 strains of fungi from the phyla Ascomycota, Basidiomycota, Chytridiomycota and Zygomycota were annotated on the genomic scale according to the classification of glycoside hydrolase (GH from the Carbohydrate-Active enZymes (CAZy Database. Comparisons of gene abundance in the fungi suggested that the repertoire of amylolytic genes adapted to their respective lifestyles. Amylolytic enzymes in family GH13 were divided into four distinct clades identified as heterologous α-amylases, eukaryotic α-amylases, bacterial and fungal α-amylases and GH13 α-glucosidases. Family GH15 had two branches, one for gluocoamylases, and the other with currently unknown function. GH31 α-glucosidases showed diverse branches consisting of neutral α-glucosidases, lysosomal acid α-glucosidases and a new clade phylogenetically related to the bacterial counterparts. Distribution of starch-binding domains in above fungal amylolytic enzymes was related to the enzyme source and phylogeny. Finally, likely scenarios for the evolution of amylolytic enzymes in fungi based on phylogenetic analyses were proposed. Our results provide new insights into evolutionary relationships among subgroups of fungal amylolytic enzymes and fungal evolutionary adaptation to ecological conditions.

  5. Preparation and characterization of room temperature ionic liquid/single-walled carbon nanotube nanocomposites and their application to the direct electrochemistry of heme-containing proteins/enzymes

    International Nuclear Information System (INIS)

    Du, Pan; Liu, Shuna; Wu, Ping; Cai, Chenxin

    2007-01-01

    This work describes the formation and possible electrochemical application of a novel nanocomposite based on single-walled carbon nanotubes (SWNTs) and imidazolium-based room-temperature ionic liquids (RTILs) of 1-butyl-3-methylimidazolium tetrafluoroborate ([bmim]BF 4 , a hydrophilic RTIL) and 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim]PF 6 , a hydrophobic RTIL). The nanocomposites ([bmim]BF 4 -SWNTs, and [bmim]PF 6 -SWNTs) were formed by simply grinding the SWNTs with the respective RTIL. The results of the X-ray photoelectron spectroscopy (XPS) and Raman spectroscopy indicated that the nanocomposites were formed by adsorption of an imidazolium ion on the surface of SWNTs via the 'cation-π' interaction. SEM images showed that [bmim]BF 4 -SWNTs (or [bmim]PF 6 -SWNTs) nanocomposites could uniformly cover the surface of a glassy carbon (GC) electrode resulting in a RTILs-SWNTs/GC modified electrode with a high stability. The RTILs-SWNTs composite could be readily used as a matrix to immobilize heme-containing proteins/enzymes (myoglobin, cytochrome c, and horseradish peroxidase) without undergoing denaturation, as was verified by UV-vis and circular dichroic (CD) spectroscopic results. The voltammetric results showed that heme-containing proteins/enzymes entrapped in RTILs-SWNTs composites displayed a pair of well-defined, stable redox peaks, which were ascribed to their direct electron-transfer reactions. The results of controlled experiments showed that the positive charged imidazolium ion played a significant effect on the electrochemical parameters, such as the redox peak separation and the value of the formal potentials, etc., of the electron-transfer reaction of non-neutral species dissolved in solution or immobilized on the electrode surface. Further results demonstrated that the heme-containing proteins/enzymes entrapped in RTILs-SWNTs composites could still retain their bioelectrocatalytic activity toward the reduction of oxygen and hydrogen

  6. Expanding the product portfolio of fungal type I fatty acid synthases

    DEFF Research Database (Denmark)

    Zhu, Zhiwei; Zhou, Yongjin J.; Krivoruchko, Anastasia

    2017-01-01

    Fungal type I fatty acid synthases (FASs) are mega-enzymes with two separated, identical compartments, in which the acyl carrier protein (ACP) domains shuttle substrates to catalytically active sites embedded in the chamber wall. We devised synthetic FASs by integrating heterologous enzymes...

  7. The importance of mass spectrometric dereplication in fungal secondary metabolite analysis

    Directory of Open Access Journals (Sweden)

    Kristian Fog Nielsen

    2015-02-01

    Full Text Available Having entered the Genomic Era, it is now evident that the biosynthetic potential of filamentous fungi is much larger than was thought even a decade ago. Fungi harbor many cryptic gene clusters encoding for the biosynthesis of polyketides, non-ribosomal peptides, and terpenoids – which can all undergo extensive modifications by tailoring enzymes – thus potentially providing a large array of products from a single pathway. Elucidating the full chemical profile of a fungal species is a challenging exercise, even with elemental composition provided by high-resolution mass spectrometry (HRMS used in combination with chemical databases (e.g. Antibase to dereplicate known compounds. This has led to a continuous effort to improve chromatographic separation in conjunction with improvement in HRMS detection. Major improvements have also occurred with 2D chromatography, ion-mobility, MS/MS and MS3, stable isotope labeling feeding experiments, classic UV/Vis, and especially automated data-mining and metabolomics software approaches as the sheer amount of data generated is now the major challenge. This review will focus on the development and implementation of dereplication strategies and will highlight the importance of each stage of the process from sample preparation to chromatographic separation and finally towards both manual and more targeted methods for automated dereplication of fungal natural products using state-of-the art MS instrumentation.

  8. 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.

  9. Fungal biodiversity to biotechnology.

    Science.gov (United States)

    Chambergo, Felipe S; Valencia, Estela Y

    2016-03-01

    Fungal habitats include soil, water, and extreme environments. With around 100,000 fungus species already described, it is estimated that 5.1 million fungus species exist on our planet, making fungi one of the largest and most diverse kingdoms of eukaryotes. Fungi show remarkable metabolic features due to a sophisticated genomic network and are important for the production of biotechnological compounds that greatly impact our society in many ways. In this review, we present the current state of knowledge on fungal biodiversity, with special emphasis on filamentous fungi and the most recent discoveries in the field of identification and production of biotechnological compounds. More than 250 fungus species have been studied to produce these biotechnological compounds. This review focuses on three of the branches generally accepted in biotechnological applications, which have been identified by a color code: red, green, and white for pharmaceutical, agricultural, and industrial biotechnology, respectively. We also discuss future prospects for the use of filamentous fungi in biotechnology application.

  10. BAKERY ENZYMES IN CEREAL TECHNOLOGIES

    Directory of Open Access Journals (Sweden)

    Václav Koman

    2012-10-01

    Full Text Available Normal 0 21 false false false SK X-NONE X-NONE Bread is the most common and traditional food in the world. For years, enzymes such as malt and fungal alpha-amylase have been used in bread making. Due to the changes in the baking industry and the ever-increasing demand for more natural products, enzymes have gained real importance in bread-making. If an enzyme is added, it is often destroyed by the heat during the baking process. For generations, enzymes have been used for the improvement of texture and appearance, enhancement of nutritional values and generation of appealing flavours and aromas. Enzymes used in bakery industry constitute nearly one third of the market. The bakery products have undergone radical improvements in quality over the past years in terms of flavour, texture and shelf-life. The the biggest contributor for these improvementsis the usage of enzymes. Present work seeks to systematically describe bakery enzymes, their classification, benefits, usage and chemical reactions in the bread making process.doi:10.5219/193

  11. Enzyme assays.

    Science.gov (United States)

    Brodelius, P E

    1991-02-01

    The past year or so has seen the development of new enzyme assays, as well as the improvement of existing ones. Assays are becoming more rapid and sensitive as a result of modifications such as amplification of the enzyme product(s). Recombinant DNA technology is now being recognized as a particularly useful tool in the search for improved assay systems.

  12. The evolution of fungal epiphytes

    NARCIS (Netherlands)

    Hongsanan, S.; Sánchez-Ramírez, S.; Crous, P.W.; Ariyawansa, H.A.; Zhao, R.L.; Hyde, K.D.

    2016-01-01

    Fungal epiphytes are a polyphyletic group found on the surface of plants, particularly on leaves, with a worldwide distribution. They belong in the phylum Ascomycota, which contains the largest known number of fungal genera. There has been little research dating the origins of the common ancestors

  13. Current management of fungal infections.

    NARCIS (Netherlands)

    Meis, J.F.G.M.; Verweij, P.E.

    2001-01-01

    The management of superficial fungal infections differs significantly from the management of systemic fungal infections. Most superficial infections are treated with topical antifungal agents, the choice of agent being determined by the site and extent of the infection and by the causative organism,

  14. Hospitalized Patients and Fungal Infections

    Science.gov (United States)

    ... Care Med 1998;24:206-16. Alangaden GJ. Nosocomial Fungal Infections: Epidemiology, Infection Control, and Prevention. Infectious Disease Clinics ... 25:201-25. Zilberberg MD, Shorr AF. Fungal infections in the ICU. Infect Dis ... D. Nosocomial aspergillosis and building construction. Med Mycol 2009;47 ...

  15. Serious fungal infections in Pakistan.

    Science.gov (United States)

    Jabeen, K; Farooqi, J; Mirza, S; Denning, D; Zafar, A

    2017-06-01

    The true burden of fungal infection in Pakistan is unknown. High-risk populations for fungal infections [tuberculosis (TB), diabetes, chronic respiratory diseases, asthma, cancer, transplant and human immunodeficiency virus (HIV) infection] are numerous. Here, we estimate the burden of fungal infections to highlight their public health significance. Whole and at-risk population estimates were obtained from the WHO (TB), BREATHE study (COPD), UNAIDS (HIV), GLOBOCAN (cancer) and Heartfile (diabetes). Published data from Pakistan reporting fungal infections rates in general and specific populations were reviewed and used when applicable. Estimates were made for the whole population or specific populations at risk, as previously described in the LIFE methodology. Of the 184,500,000 people in Pakistan, an estimated 3,280,549 (1.78%) are affected by a serious fungal infection, omitting all cutaneous infection, oral candidiasis and allergic fungal sinusitis, which we could not estimate. Compared with other countries, the rates of candidaemia (21/100,000) and mucormycosis (14/100,000) are estimated to be very high, and are based on data from India. Chronic pulmonary aspergillosis rates are estimated to be high (39/100,000) because of the high TB burden. Invasive aspergillosis was estimated to be around 5.9/100,000. Fungal keratitis is also problematic in Pakistan, with an estimated rate of 44/100,000. Pakistan probably has a high rate of certain life- or sight-threatening fungal infections.

  16. SHORT COMMUNICATION ANTI-FUNGAL ACTIVITIES OF m ...

    African Journals Online (AJOL)

    a

    of these metals complexes as food preservatives may only be fungi-static and not fungi-toxic, their use in bread preparation might extend the shelf life of bread from 24 hours to 96 hours. KEY WORDS: Anti-fungal activities, Alkali metal iodobenzoates, Alkaline earth metal iodobenzoates, m-Iodobenzoic acid, Bread mucor.

  17. Blue light (470 nm) effectively inhibits bacterial and fungal growth

    Science.gov (United States)

    The activity of blue light (470nm) alone on (1) bacterial viability, and (2) with a food grade photosensitizer on filamentous fungal viability, was studied. Suspensions of the bacteria Leuconostoc mesenteroides (LM), Bacillus atrophaeus (BA), and Pseudomonas aeruginosa (PA) were prepared and aliquo...

  18. Preparation of riboflavin specifically labeled in the 5'-hydroxymethyl terminus using a vitamin B2-aldehyde-forming enzyme from Schizophyllum commune

    International Nuclear Information System (INIS)

    Kekelidze, T.N.; Edmondson, D.E.; McCormick, D.B.

    1995-01-01

    A method is described for synthesis of riboflavin selectively labeled in the hydrogens at the 5'-hydroxymethyl position. In this method, a vitamin B 2 -aldehyde-forming enzyme from Schizophyllum commune is used to specifically and completely oxidize the 5'-hydroxymethyl of riboflavin to the 5'-aldehyde. This reaction is monitored spectrophotometrically by the reduction of 2,6-dichlorophenolindophenol at 600 nm. Appearance of aldehyde product was directly quantitated by reverse-phase high-performance liquid chromatography. Product is extracted from the incubation mixture by phenol after saturation with (NH 4 ) 2 SO 4 and then further purified by benzyl alcohol extraction. The 5'-aldehyde is reduced with appropriately labeled sodium borohydride to yield the vitamin specifically labeled in the 5'-hydroxymethyl group. (author)

  19. Production of plant cell wall degrading enzymes by monoculture and co-culture of Aspergillus niger and Aspergillus terreus under SSF of banana peels

    Directory of Open Access Journals (Sweden)

    Shazia Rehman

    2014-12-01

    Full Text Available Filamentous fungi are considered to be the most important group of microorganisms for the production of plant cell wall degrading enzymes (CWDE, in solid state fermentations. In this study, two fungal strains Aspergillus niger MS23 and Aspergillus terreus MS105 were screened for plant CWDE such as amylase, pectinase, xylanase and cellulases (β-glucosidase, endoglucanase and filterpaperase using a novel substrate, Banana Peels (BP for SSF process. This is the first study, to the best of our knowledge, to use BP as SSF substrate for plant CWDE production by co-culture of fungal strains. The titers of pectinase were significantly improved in co-culture compared to mono-culture. Furthermore, the enzyme preparations obtained from monoculture and co-culture were used to study the hydrolysis of BP along with some crude and purified substrates. It was observed that the enzymatic hydrolysis of different crude and purified substrates accomplished after 26 h of incubation, where pectin was maximally hydrolyzed by the enzyme preparations of mono and co-culture. Along with purified substrates, crude materials were also proved to be efficiently degraded by the cocktail of the CWDE. These results demonstrated that banana peels may be a potential substrate in solid-state fermentation for the production of plant cell wall degrading enzymes to be used for improving various biotechnological and industrial processes.

  20. Ligninolytic enzyme activities in mycelium of some wild and ...

    African Journals Online (AJOL)

    Lignin is probably one of the most recalcitrant compounds synthesized by plants. This compound is degraded by few microorganisms. White-rot fungi have been extensively studied due to its powerful ligninolytic enzymes. In this study, ligninolytic enzyme activities of different fungal species (six commercial and 13 wild) were ...

  1. Ligninolytic enzyme activities in mycelium of some wild and ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-12-03

    Dec 3, 2008 ... Lignin is probably one of the most recalcitrant compounds synthesized by plants. This compound is degraded by few microorganisms. White-rot fungi have been extensively studied due to its powerful ligninolytic enzymes. In this study, ligninolytic enzyme activities of different fungal species (six commercial ...

  2. Engineering Foundation Conference on Enzyme Engineering XII

    National Research Council Canada - National Science Library

    Russell, Author

    1997-01-01

    Partial Contents: Preparation and Properties of Designed Biocatalysts Biopolymer Structure and Function Biocatalysts under Extreme Environments Application of Protein Expression in Biocatalysis Biochemical Engineering of Enzyme Systems...

  3. Hydrolytic enzymes in the central vacuole of plant cells

    International Nuclear Information System (INIS)

    Boller, T.; Kende, H.

    1979-01-01

    The hydrolase content of vacuoles isolated from protoplasts of suspension-cultured tobacco cells, of tulip petals, and pineapple leaves, and the sedimentation behavior of tobacco tonoplasts were studied. Three precautions were found to be important for the analysis of vacuolar hydrolases and of the tonoplast: (a) purification of protoplasts in a Ficoll gradient was necessary to remove cell debris which contained contaminating hydrolases adsorbed from the fungal cell-wall-degrading enzyme preparation; (b) hydrolase activities in the homogenates of the intact cells or the tissue used and of the purified protoplasts had to be compared to verify the absence of contaminating hydrolases in the protoplast preparation; and (c) vacuoles obtained from the protoplasts by an osmotic shock had to be purified from the lysate in a Ficoll gradient. Since the density of the central vacuole approximates that of the protoplasts, about a 10% contamination of the vacuolar preparation by surviving protoplasts could not be eliminated. The intracellular activities of the following acid hydrolases were primarily localized in the vacuole of tobacco cells: α-mannosidase, β-N-acetylglucosaminidase, β-fructosidase, nuclease, phosphatase, phosphodiesterase. A similar composition of acid hydrolases was found in vacuoles obtained from protoplasts of tulip petals. Proteinase, a hydrolase with low activity in tobacco cells and tulip petals was found to be vacuolar in pineapple leaves, a tissue containing high levels of this enzyme. None of the vacuolar enzymes investigated ws found to be bound to the tonoplast. When vacuoles were isolated from cells labeled with radioactive choline, the vacuolar membrane was found to contain radioactivity. On sucrose gradients, the label incorporated into tonoplasts banded around a density of 1.10 grams per cubic centimeter

  4. Friends or foes? Emerging insights from fungal interactions with plants

    Science.gov (United States)

    Zeilinger, Susanne; Gupta, Vijai K.; Dahms, Tanya E. S.; Silva, Roberto N.; Singh, Harikesh B.; Upadhyay, Ram S.; Gomes, Eriston Vieira; Tsui, Clement Kin-Ming; Nayak S, Chandra

    2015-01-01

    Fungi interact with plants in various ways, with each interaction giving rise to different alterations in both partners. While fungal pathogens have detrimental effects on plant physiology, mutualistic fungi augment host defence responses to pathogens and/or improve plant nutrient uptake. Tropic growth towards plant roots or stomata, mediated by chemical and topographical signals, has been described for several fungi, with evidence of species-specific signals and sensing mechanisms. Fungal partners secrete bioactive molecules such as small peptide effectors, enzymes and secondary metabolites which facilitate colonization and contribute to both symbiotic and pathogenic relationships. There has been tremendous advancement in fungal molecular biology, omics sciences and microscopy in recent years, opening up new possibilities for the identification of key molecular mechanisms in plant–fungal interactions, the power of which is often borne out in their combination. Our fragmentary knowledge on the interactions between plants and fungi must be made whole to understand the potential of fungi in preventing plant diseases, improving plant productivity and understanding ecosystem stability. Here, we review innovative methods and the associated new insights into plant–fungal interactions. PMID:26591004

  5. Navigating the fungal polyketide chemical space: from genes to molecules.

    Science.gov (United States)

    Chooi, Yit-Heng; Tang, Yi

    2012-11-16

    The iterative type I polyketide synthases (IPKSs) are central to the biosynthesis of an enormously diverse array of natural products in fungi. These natural products, known as polyketides, exhibit a wide range of biological activities and include clinically important drugs as well as undesirable toxins. The PKSs synthesize these structurally diverse polyketides via a series of decarboxylative condensations of malonyl-CoA extender units and β-keto modifications in a highly programmed manner. Significant progress has been made over the past few years in understanding the biosynthetic mechanism and programming of fungal PKSs. The continuously expanding fungal genome sequence data have sparked genome-directed discoveries of new fungal PKSs and associated products. The increasing number of fungal PKSs that have been linked to their products along with in-depth biochemical and structural characterizations of these large enzymes have remarkably improved our knowledge on the molecular basis for polyketide structural diversity in fungi. This Perspective highlights the recent advances and examines how the newly expanded paradigm has contributed to our ability to link fungal PKS genes to chemical structures and vice versa. The knowledge will help us navigate through the logarithmically expanding seas of genomic information for polyketide compound discovery and provided opportunities to reprogram these megasynthases to generate new chemical entities.

  6. Friends or foes? Emerging insights from fungal interactions with plants.

    Science.gov (United States)

    Zeilinger, Susanne; Gupta, Vijai K; Dahms, Tanya E S; Silva, Roberto N; Singh, Harikesh B; Upadhyay, Ram S; Gomes, Eriston Vieira; Tsui, Clement Kin-Ming; Nayak S, Chandra

    2016-03-01

    Fungi interact with plants in various ways, with each interaction giving rise to different alterations in both partners. While fungal pathogens have detrimental effects on plant physiology, mutualistic fungi augment host defence responses to pathogens and/or improve plant nutrient uptake. Tropic growth towards plant roots or stomata, mediated by chemical and topographical signals, has been described for several fungi, with evidence of species-specific signals and sensing mechanisms. Fungal partners secrete bioactive molecules such as small peptide effectors, enzymes and secondary metabolites which facilitate colonization and contribute to both symbiotic and pathogenic relationships. There has been tremendous advancement in fungal molecular biology, omics sciences and microscopy in recent years, opening up new possibilities for the identification of key molecular mechanisms in plant-fungal interactions, the power of which is often borne out in their combination. Our fragmentary knowledge on the interactions between plants and fungi must be made whole to understand the potential of fungi in preventing plant diseases, improving plant productivity and understanding ecosystem stability. Here, we review innovative methods and the associated new insights into plant-fungal interactions. © FEMS 2015.

  7. An investigation on non-invasive fungal sinusitis; Molecular identification of etiologic agents

    Directory of Open Access Journals (Sweden)

    Abdolrasoul Mohammadi

    2017-01-01

    Full Text Available Background: Fungal sinusitis is increasing worldwide in the past two decades. It is divided into two types including invasive and noninvasive. Noninvasive types contain allergic fungal sinusitis (AFS and fungus ball. AFS is a hypersensitivity reaction to fungal allergens in the mucosa of the sinonasal tract in atopic individuals. The fungus ball is a different type of noninvasive fungal rhinosinusitis which is delineated as an accumulation of debris and fungal elements inside a paranasal sinus. Fungal sinusitis caused by various fungi such as Aspergillus species, Penicillium, Mucor, Rhizopus, and phaeohyphomycetes. The aim of the present study is to identify fungal species isolated from noninvasive fungal sinusitis by molecular methods. Materials and Methods: During 2015–2016, a total of 100 suspected patients were examined for fungal sinusitis. Functional endoscopic sinus surgery was performed using the Messerklinger technique. Clinical samples were identified by phenotypic and molecular methods. Polymerase chain reaction (PCR sequencing of ITS1-5.8S-ITS2 region and PCR-restriction fragment length polymorphism with Msp I restriction enzyme was performed for molecular identification of molds and yeasts, respectively. Results: Twenty-seven out of 100 suspected cases (27% had fungal sinusitis. Nasal congestion (59% and headache (19% were the most common clinical signs among patients. Fifteen patients (55.5% were male and 12 patients (44.5% were female. Aspergillus flavus was the most prevalent fungal species (26%, followed by Penicillium chrysogenum (18.5% and Candida glabrata species complex (15%. Conclusion: Since clinical manifestations, computed tomography scan, endoscopy, and histopathological findings are very nonspecific in AFS and fungus ball; therefore, molecular investigations are compulsory for precise identification of etiologic agents and appropriate management of these fungal infections.

  8. [Structure and Activity of Fungal Lipases in Bile Salt Solutions].

    Science.gov (United States)

    Bogdanova, L R; Bakirova, D R; Valiullina, Yu A; Idiyatullin, B Z; Faizullin, D A; Zueva, O S; Zuev, Yu F

    2016-01-01

    The changes in structure and catalytic properties of fungal lipases (Candida rugosa, Rhizomucor miehei, Mucor javanicus) were investigated in micellar solutions of bile salts that differ in hydrophilic-lypophilic balance and reaction medium properties. The methods of circular dichroism and tryptophan fluorescence were applied to estimate the changes in peptide structure within complexes with bile salt micelles. Bile salts do not exert a significant influence on the structure of the enzymes under study: in Rh. miehei and M. javanicus lipases the alpha helix content slightly decreased, the influence of bile salts on the C. rugosa structure was not revealed. Despite negligible structural modifications in the enzymes, in bile salt solutions a considerable change in their catalytic properties was observed: an abrupt decrease in catalytic effectiveness. Substrate-bile salts micelles complex formation was demonstrated by the NMR self-diffusion method. The model of a regulation of fungal lipase activity was proposed.

  9. Fungal elicitors of the phytoalexin response in higher plants

    Science.gov (United States)

    West, Charles A.

    1981-09-01

    Several types of fungal molecules including cell wall polysaccharides, polypeptides, glycoproteins and lipid molecules have been found to serve as elicitors of phytoalexins in higher plants. Recent work has shown that an extracellular enzyme, endopolygalacturonase, from culture filtrates of the fungus Rhizopus stolonifer elicits the biosynthesis of an antifungal antibiotic, casbene, in extracts of treated castor bean ( Ricinus communis L.) seedlings. A suggested mode of action of this elicitor in the plant in which fragments of the plant cell wall released through the catalytic action of the enzyme serve as secondary elicitors to trigger the plant response is proposed on the basis of preliminary observations. Possible modes of interaction of other types of fungal elicitors with plants are also discussed.

  10. Strategies to Characterize Fungal Lipases for Applications in Medicine and Dairy Industry

    Directory of Open Access Journals (Sweden)

    Subash C. B. Gopinath

    2013-01-01

    Full Text Available Lipases are water-soluble enzymes that act on insoluble substrates and catalyze the hydrolysis of long-chain triglycerides. Lipases play a vital role in the food, detergent, chemical, and pharmaceutical industries. In the past, fungal lipases gained significant attention in the industries due to their substrate specificity and stability under varied chemical and physical conditions. Fungal enzymes are extracellular in nature, and they can be extracted easily, which significantly reduces the cost and makes this source preferable over bacteria. Soil contaminated with spillage from the products of oil and dairy harbors fungal species, which have the potential to secrete lipases to degrade fats and oils. Herein, the strategies involved in the characterization of fungal lipases, capable of degrading fatty substances, are narrated with a focus on further applications.

  11. Strategies to Characterize Fungal Lipases for Applications in Medicine and Dairy Industry

    Science.gov (United States)

    Gopinath, Subash C. B.; Anbu, Periasamy; Lakshmipriya, Thangavel; Hilda, Azariah

    2013-01-01

    Lipases are water-soluble enzymes that act on insoluble substrates and catalyze the hydrolysis of long-chain triglycerides. Lipases play a vital role in the food, detergent, chemical, and pharmaceutical industries. In the past, fungal lipases gained significant attention in the industries due to their substrate specificity and stability under varied chemical and physical conditions. Fungal enzymes are extracellular in nature, and they can be extracted easily, which significantly reduces the cost and makes this source preferable over bacteria. Soil contaminated with spillage from the products of oil and dairy harbors fungal species, which have the potential to secrete lipases to degrade fats and oils. Herein, the strategies involved in the characterization of fungal lipases, capable of degrading fatty substances, are narrated with a focus on further applications. PMID:23865040

  12. Recommendations for Risk Categorization and Prophylaxis of Invasive Fungal Diseases in Hematological Malignancies: A Critical Review of Evidence and Expert Opinion (TEO-4

    Directory of Open Access Journals (Sweden)

    Can Boğa

    2015-06-01

    Full Text Available This is the last of a series of articles on invasive fungal infections prepared by opinion leaders in Turkey. The aim of these articles is to guide clinicians in managing invasive fungal diseases in hematological malignancies and stem cell transplantation based on the available best evidence in this field. The previous articles summarized the diagnosis and treatment of invasive fungal disease and this article aims to explain the risk categorization and guide the antifungal prophylaxis in invasive fungal disease.

  13. Preparation, crystallization and X-ray diffraction analysis to 1.5 Å resolution of rat cysteine dioxygenase, a mononuclear iron enzyme responsible for cysteine thiol oxidation

    Energy Technology Data Exchange (ETDEWEB)

    Simmons, Chad R. [Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853-8001 (United States); Hao, Quan [MacCHESS at the Cornell High Energy Synchrotron Source, Cornell University, Ithaca, NY 14853-8001 (United States); Stipanuk, Martha H., E-mail: mhs6@cornell.edu [Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853-8001 (United States)

    2005-11-01

    Recombinant rat cysteine dioxygenase (CDO) has been expressed, purified and crystallized and X-ray diffraction data have been collected to 1.5 Å resolution. Cysteine dioxygenase (CDO; EC 1.13.11.20) is an ∼23 kDa non-heme iron metalloenzyme that is responsible for the oxidation of cysteine by O{sub 2}, yielding cysteinesulfinate. CDO catalyzes the first step in the conversion of cysteine to taurine, as well as the first step in the catabolism of cysteine to pyruvate plus sulfate. Recombinant rat CDO was heterologously expressed, purified and crystallized. The protein was expressed as a fusion protein bearing a polyhistidine tag to facilitate purification, a thioredoxin tag to improve solubility and a factor Xa cleavage site to permit removal of the entire N-terminus, leaving only the 200 amino acids inherent to the native protein. A multi-step purification scheme was used to achieve >95% purity of CDO. The optimal CDO crystals diffracted to 1.5 Å resolution and belonged to space group P4{sub 3}2{sub 1}2 or P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 57.55, c = 123.06 Å, α = β = γ = 90°. CDO shows little homology to any other proteins; therefore, the structure of the enzyme will be determined by ab initio phasing using a selenomethionyl derivative.

  14. Preparation, Crystallization and X-ray Diffraction Analysis to 1.5 A Resolution of Rat Cysteine Dioxygenase, a Mononuclear Iron Enzyme Responsible for Cysteine Thiol Oxidation

    Energy Technology Data Exchange (ETDEWEB)

    Simmons,C.; Hao, Q.; Stipanuk, M.

    2005-01-01

    Cysteine dioxygenase (CDO; EC 1.13.11.20) is an {approx}23 kDa non-heme iron metalloenzyme that is responsible for the oxidation of cysteine by O2, yielding cysteinesulfinate. CDO catalyzes the first step in the conversion of cysteine to taurine, as well as the first step in the catabolism of cysteine to pyruvate plus sulfate. Recombinant rat CDO was heterologously expressed, purified and crystallized. The protein was expressed as a fusion protein bearing a polyhistidine tag to facilitate purification, a thioredoxin tag to improve solubility and a factor Xa cleavage site to permit removal of the entire N-terminus, leaving only the 200 amino acids inherent to the native protein. A multi-step purification scheme was used to achieve >95% purity of CDO. The optimal CDO crystals diffracted to 1.5 Angstroms resolution and belonged to space group P4{sub 3}2{sub 1}2 or P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 57.55, c = 123.06 Angstrom, {alpha} = {beta} = {gamma} = 90. CDO shows little homology to any other proteins; therefore, the structure of the enzyme will be determined by ab initio phasing using a selenomethionyl derivative.

  15. Dynamics of proteolytic activity of blood enzymes in combined burn and radiation injury and in its treatment with local cryoeffect and with wound closing preparation RZP-3

    International Nuclear Information System (INIS)

    Gertman, V.Z.

    1982-01-01

    The proteolytic activity of trypsine-like proteases of blood at acute injury period doesn't increase as in animals with mere burn and in animals with nure irradiation as well. It can be explained by the absence of adequate stress reaction of the organism due to the sharp decrease of organism reactivity in case of combined burn and radiation injury. Application of low temperatures and combination of cryogenic effect and RZP-3 preparation promotes proteolytic activity increase in animals with combined burn and radiation injury in the climax of radiation sickness. The normalization of the factor was observed at late periods of the investigation. It can be regarded as recovery of the organism reactivity

  16. New insights into the formation of fungal aromatic polyketides

    OpenAIRE

    Crawford, Jason M.; Townsend, Craig A.

    2010-01-01

    Fungal aromatic polyketides constitute a large family of bioactive natural products and are synthesized by the non-reducing group of iterative polyketide synthases (NR-PKSs). Their diverse structures arise from selective enzymatic modifications of reactive enzyme-bound poly-β-keto intermediates. How iterative PKSs control starter unit selection, polyketide chain initiation and elongation, intermediate folding and cyclization, selective redox or modification reactions during assembly, and prod...

  17. Widespread Occurrence of Expressed Fungal Secretory Peroxidases in Forest Soils

    OpenAIRE

    Kellner, Harald; Luis, Patricia; Pecyna, Marek J.; Barbi, Florian; Kapturska, Danuta; Krüger, Dirk; Zak, Donald R.; Marmeisse, Roland; Vandenbol, Micheline; Hofrichter, Martin

    2014-01-01

    Fungal secretory peroxidases mediate fundamental ecological functions in the conversion and degradation of plant biomass. Many of these enzymes have strong oxidizing activities towards aromatic compounds and are involved in the degradation of plant cell wall (lignin) and humus. They comprise three major groups: class II peroxidases (including lignin peroxidase, manganese peroxidase, versatile peroxidase and generic peroxidase), dye-decolorizing peroxidases, and heme-thiolate peroxidases (e.g....

  18. Effect of polyols on fungal alpha-amylase thermostability

    OpenAIRE

    Graber, Marianne; Combes, Didier

    1989-01-01

    International audience; The influence of different polyols (ethylene glycol, glycerol, erythritol, xylitol, and sorbitol) on the thermostability of fungal alpha-amylase at 60°C has been studied. The results obtained show a stabilizing effect in the presence of polyols. In the case of 4 m sorbitol solution, the enzyme half-life is 2000-fold longer than in pure water. These polyols have been found as competitive inhibitors for alpha-amylase andtheir stabilizing effect has been correlated to the...

  19. Combining regio- and enantioselectivity of lipases for the preparation of (R)-4-chloro-2-butanol.

    Science.gov (United States)

    Méndez, Jonh J; Oromi, Mireia; Cervero, Maria; Balcells, Mercè; Torres, Mercè; Canela, Ramon

    2007-01-01

    Preparation of 98% ee (R)-4-chloro-2-butanol was carried out by the enzymatic hydrolysis of chlorohydrin esters, using fungal resting cells and commercial enzymes. Hydrolyzes were carried out using lipases from Candida antarctica (Novozym 435), C. rugosa, Rhizomucor miehei (Lipozyme IM), Burkolia cepacia, and resting cells of Rhizopus oryzae and Aspergillus flavus. The influence of the enzyme, the solvent, the temperature, and the alkyl chain length on the selectivity of hydrolyzes of isomeric mixtures of chlorohydrin esters is described. Regioselectivity was higher than 95% for some of the tested lipases. Novozym 435 allowed preparation of the (R)-4-chloro-2-butanol after 15 min of reaction at 30-40 degrees C. (c) 2006 Wiley-Liss, Inc.

  20. Preparation and application of unhairing enzyme using solid wastes from the leather industry-an attempt toward internalization of solid wastes within the leather industry.

    Science.gov (United States)

    Ramesh, Renganath Rao; Muralidharan, Vimudha; Palanivel, Saravanan

    2018-01-01

    Usage of the animal fleshing waste as the source of carbon and nitrogen for animal skin unhairing protease (EC 3.4.21) production along with agro-industrial wastes like wheat bran has been investigated. Thermal hydrolysis of delimed fleshing waste for 3 h yielded a fleshing hydrolysate (FH) having a protein content of 20.86 mg/mL and total solids of 46,600 ppm. The FH was lyophilized and spray dried to obtain fleshing hydrolysate powder (FHP) to be used along with wheat bran and rice bran for protease production. The carbon, nitrogen, hydrogen, and sulfur contents of the FHP were found to be 40.1, 13.8, 5.4, and 0.2%. The control solid-state fermented (SSF) medium without FHP showed a maximum activity of only 550 U/g. A maximum protease activity of 956 U/g was obtained by using 6% FHP (taken based on the combined total weight of wheat bran and rice bran) after 96 h of fermentation, resulting in a 1.7-fold increase in the protease activity. The total cost of producing 1 kg of FHP and the cost of producing 1000 kU of protease using FHP along with wheat bran and rice bran were found to be USD 24.62 and USD 2.08, respectively; 25% of SSF protease along with 40% water was found to be capable of unhairing the sheepskins in 7 h eliminating the hazardous conventional lime sulfide unhairing system. Thus, the leather industry's solid waste internalized for the production of unhairing enzyme resulted in a sustainable solution for pollution problems. Graphical abstract ᅟ.

  1. Fungal laccases degradation of endocrine disrupting compounds.

    Science.gov (United States)

    Macellaro, Gemma; Pezzella, Cinzia; Cicatiello, Paola; Sannia, Giovanni; Piscitelli, Alessandra

    2014-01-01

    Over the past decades, water pollution by trace organic compounds (ng/L) has become one of the key environmental issues in developed countries. This is the case of the emerging contaminants called endocrine disrupting compounds (EDCs). EDCs are a new class of environmental pollutants able to mimic or antagonize the effects of endogenous hormones, and are recently drawing scientific and public attention. Their widespread presence in the environment solicits the need of their removal from the contaminated sites. One promising approach to face this challenge consists in the use of enzymatic systems able to react with these molecules. Among the possible enzymes, oxidative enzymes are attracting increasing attention because of their versatility, the possibility to produce them on large scale, and to modify their properties. In this study five different EDCs were treated with four different fungal laccases, also in the presence of both synthetic and natural mediators. Mediators significantly increased the efficiency of the enzymatic treatment, promoting the degradation of substrates recalcitrant to laccase oxidation. The laccase showing the best performances was chosen to further investigate its oxidative capabilities against micropollutant mixtures. Improvement of enzyme performances in nonylphenol degradation rate was achieved through immobilization on glass beads.

  2. Fungal Laccases Degradation of Endocrine Disrupting Compounds

    Directory of Open Access Journals (Sweden)

    Gemma Macellaro

    2014-01-01

    Full Text Available Over the past decades, water pollution by trace organic compounds (ng/L has become one of the key environmental issues in developed countries. This is the case of the emerging contaminants called endocrine disrupting compounds (EDCs. EDCs are a new class of environmental pollutants able to mimic or antagonize the effects of endogenous hormones, and are recently drawing scientific and public attention. Their widespread presence in the environment solicits the need of their removal from the contaminated sites. One promising approach to face this challenge consists in the use of enzymatic systems able to react with these molecules. Among the possible enzymes, oxidative enzymes are attracting increasing attention because of their versatility, the possibility to produce them on large scale, and to modify their properties. In this study five different EDCs were treated with four different fungal laccases, also in the presence of both synthetic and natural mediators. Mediators significantly increased the efficiency of the enzymatic treatment, promoting the degradation of substrates recalcitrant to laccase oxidation. The laccase showing the best performances was chosen to further investigate its oxidative capabilities against micropollutant mixtures. Improvement of enzyme performances in nonylphenol degradation rate was achieved through immobilization on glass beads.

  3. Chitinase production by Streptomyces viridificans: its potential in fungal cell wall lysis.

    Science.gov (United States)

    Gupta, R; Saxena, R K; Chaturvedi, P; Virdi, J S

    1995-04-01

    Streptomyces viridificans was found to be a good chitinase producer among nine species of Streptomyces screened. Minimum levels of constitutive enzyme were observed with both simple and complex carbon substrate. Arabinose doubled the enzyme production amongst the various pentoses and hexoses used with chitin. However, with glucose end-product inhibition and catabolite repression were observed. The enzyme tolerated a wide range of temperature (30-55 degrees C) and pH (3-7.5). Among various divalent cations Mn2+ and Hg2+ completely inhibited the purified enzyme while beta-mercaptoethanol stimulated its activity. Crude and purified enzyme had potential for cell wall lysis of many fungal pathogens tested.

  4. Enzymatic bioremediation of polyaromatic hydrocarbons by fungal consortia enriched from petroleum contaminated soil and oil seeds.

    Science.gov (United States)

    Balaji, V; Arulazhagan, P; Ebenezer, P

    2014-05-01

    The present study focuses on fungal strains capable of secreting extracellular enzymes by utilizing hydrocarbons present in the contaminated soil. Fungal strains were enriched from petroleum hydrocarbons contaminated soil samples collected from Chennai city, India. The potential fungi were isolated and screened for their enzyme secretion such as lipase, laccase, peroxidase and protease and also evaluated fungal enzyme mediated PAHs degradation. Total, 21 potential PAHs degrading fungi were isolated from PAHs contaminated soil, which belongs to 9 genera such as Aspergillus, Curvularia, Drechslera, Fusarium, Lasiodiplodia, Mucor Penicillium, Rhizopus, Trichoderma, and two oilseed-associated fungal genera such as Colletotrichum and Lasiodiplodia were used to test their efficacy in degradation of PAHs in polluted soil. Maximum lipase production was obtained with P. chrysogenum, M. racemosus and L. theobromae VBE1 under optimized cultural condition, which utilized PAHs in contaminated soil as sole carbon source. Fungal strains, P. chrysogenum, M. racemosus and L. theobromae VBE1, as consortia, used in the present study were capable of degrading branched alkane isoprenoids such as pristine (C17) and pyrene (C18) present in PAHs contaminated soil with high lipase production. The fungal consortia acts as potential candidate for bioremediation of PAHs contaminated environments.

  5. Fungal proteomics: from identification to function.

    Science.gov (United States)

    Doyle, Sean

    2011-08-01

    Some fungi cause disease in humans and plants, while others have demonstrable potential for the control of insect pests. In addition, fungi are also a rich reservoir of therapeutic metabolites and industrially useful enzymes. Detailed analysis of fungal biochemistry is now enabled by multiple technologies including protein mass spectrometry, genome and transcriptome sequencing and advances in bioinformatics. Yet, the assignment of function to fungal proteins, encoded either by in silico annotated, or unannotated genes, remains problematic. The purpose of this review is to describe the strategies used by many researchers to reveal protein function in fungi, and more importantly, to consolidate the nomenclature of 'unknown function protein' as opposed to 'hypothetical protein' - once any protein has been identified by protein mass spectrometry. A combination of approaches including comparative proteomics, pathogen-induced protein expression and immunoproteomics are outlined, which, when used in combination with a variety of other techniques (e.g. functional genomics, microarray analysis, immunochemical and infection model systems), appear to yield comprehensive and definitive information on protein function in fungi. The relative advantages of proteomic, as opposed to transcriptomic-only, analyses are also described. In the future, combined high-throughput, quantitative proteomics, allied to transcriptomic sequencing, are set to reveal much about protein function in fungi. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  6. Human plasma lecithin:cholesterol acyltransferase. Preparation and use of immobilized p-aminophenylarsenoxide as a catalytic site-directed covalent ligand in enzyme purification.

    Science.gov (United States)

    Zhou, G Y; Jauhiainen, M; Stevenson, K; Dolphin, P J

    1991-07-17

    A method is described for the preparation of p-aminophenylarsenoxide-linked carboxymethyl (CM) Bio-Gel A and its use as a specific, catalytic site-directed affinity chromatography ligand in the final stages of the purification of human plasma lecithin:cholesterol acyltransferase (LCAT) (EC 2.3.1.43). Previous mechanistic studies by us demonstrated that phenylarsenoxide derivatives, which are highly specific for vicinal thiols, could inhibit LCAT via a covalent interaction with the sulphydryl groups of the two catalytic cysteine residues and that this inhibition could be rapidly and completely reversed upon addition of 2,3-dimercaptopropanesulphonic acid. The ligand was covalently linked to CM Bio-Gel A activated through an N-hydroxysuccinyl ester formed by N-hydroxysuccinimide and dicyclohexylcarbodiimide in dry dimethyl sulphoxide; 87% of the added p-aminophenylarsenoxide was coupled to the CM Bio-Gel A in 3 h at 25 degrees C giving 2.3 mg of p-aminophenylarsenoxide per ml of gel. Homogeneous LCAT free of apo A-I, apo E, apo D and albumin was obtained in an 11% yield and purified 15,013-fold overall. A 13-fold purification was obtained by chromatography upon p-aminophenylarsenoxide-CM Bio-Gel A. This method is a useful final step in LCAT purification and will prove valuable in the purification of other proteins and compounds containing vicinal thiols.

  7. DFVF: database of fungal virulence factors

    Science.gov (United States)

    Lu, Tao; Yao, Bo; Zhang, Chi

    2012-01-01

    Fungal pathogens cause various diseases for plant and animal hosts. Despite the extensive impact of fungi on human health and life, the threats posed by emerging fungal pathogens are poorly understood. Specifically, there exist few fungal virulence gene databases, which prevent effective bioinformatics studies on fungal pathogens. Therefore, we constructed a comprehensive online database of known fungal virulence factors, which collected 2058 pathogenic genes produced by 228 fungal strains from 85 genera. This database creates a pivotal platform capable of stimulating and facilitating further bench studies on fungal pathogens. Database URL: http://sysbio.unl.edu/DFVF/ PMID:23092926

  8. Cancer Patients and Fungal Infections

    Science.gov (United States)

    ... includes places like chicken coops and caves. Wear gloves when handling materials such as soil, moss, or ... MMWR: Recommendations and Reports 2000;49:1-128. Top of Page Related Links Fungal Meningitis National Center ...

  9. 21 CFR 184.1287 - Enzyme-modified fats.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Enzyme-modified fats. 184.1287 Section 184.1287... Listing of Specific Substances Affirmed as GRAS § 184.1287 Enzyme-modified fats. (a) Enzyme-modified refined beef fat, enzyme-modified butterfat, and enzyme-modified steam-rendered chicken fat are prepared...

  10. Fungal microbiota dysbiosis in IBD

    Science.gov (United States)

    Sokol, Harry; Leducq, Valentin; Aschard, Hugues; Pham, Hang-Phuong; Jegou, Sarah; Landman, Cecilia; Cohen, David; Liguori, Giuseppina; Bourrier, Anne; Nion-Larmurier, Isabelle; Cosnes, Jacques; Seksik, Philippe; Langella, Philippe; Skurnik, David; Richard, Mathias L; Beaugerie, Laurent

    2017-01-01

    Objective The bacterial intestinal microbiota plays major roles in human physiology and IBDs. Although some data suggest a role of the fungal microbiota in IBD pathogenesis, the available data are scarce. The aim of our study was to characterise the faecal fungal microbiota in patients with IBD. Design Bacterial and fungal composition of the faecal microbiota of 235 patients with IBD and 38 healthy subjects (HS) was determined using 16S and ITS2 sequencing, respectively. The obtained sequences were analysed using the Qiime pipeline to assess composition and diversity. Bacterial and fungal taxa associated with clinical parameters were identified using multivariate association with linear models. Correlation between bacterial and fungal microbiota was investigated using Spearman's test and distance correlation. Results We observed that fungal microbiota is skewed in IBD, with an increased Basidiomycota/Ascomycota ratio, a decreased proportion of Saccharomyces cerevisiae and an increased proportion of Candida albicans compared with HS. We also identified disease-specific alterations in diversity, indicating that a Crohn's disease-specific gut environment may favour fungi at the expense of bacteria. The concomitant analysis of bacterial and fungal microbiota showed a dense and homogenous correlation network in HS but a dramatically unbalanced network in IBD, suggesting the existence of disease-specific inter-kingdom alterations. Conclusions Besides bacterial dysbiosis, our study identifies a distinct fungal microbiota dysbiosis in IBD characterised by alterations in biodiversity and composition. Moreover, we unravel here disease-specific inter-kingdom network alterations in IBD, suggesting that, beyond bacteria, fungi might also play a role in IBD pathogenesis. PMID:26843508

  11. Evaluation of two Taenia solium cysticercal antigenic preparations (vesicular fluid and a glycoprotein fraction with affinity for lentil lectin for the immunodiagnosis of neurocysticercosis by enzyme-linked immunosorbent assay (ELISA

    Directory of Open Access Journals (Sweden)

    Lisandra Akemi Suzuki

    2011-06-01

    Full Text Available OBJECTIVE: To evaluate the performance of two antigenic preparations (vesicular fluid - VF and a glycoprotein fraction, LLa-Gp fraction, purified from a whole parasite extract by lentil lectin affinity chromatography from Taenia solium cysticerci for the immunodiagnosis of neurocysticercosis. METHOD: Fifty-six cerebrospinal fluid (CSF samples (22 from patients with neurocysticercosis and 34 from patients with other neurological disorders and 57 serum samples (22 from patients with neurocysticercosis, 18 from patients with other infections and 17 from presumably healthy persons were assayed for anticysticercal IgG antibodies with an enzyme-linked immunosorbent assay (ELISA. RESULTS: The VF ELISA showed 100% sensitivity and specificity in CSF and serum samples, whereas the sensitivity and specificity of the LLa-Gp ELISA were, respectively, 90.9% and 97.1%, with the CSF samples and 95.5% and 100% with serum samples. There was no significant difference in the sensitivity and specificity of the two antigenic preparations used to screen CSF and serum samples. CONCLUSION: Considering the complexity and high cost of obtaining the LLa-Gp fraction, VF could be more suitable for screening specific antibodies by ELISA in CSF and serum samples from patients with neurocysticercosis.

  12. Serious fungal infections in Ecuador.

    Science.gov (United States)

    Zurita, J; Denning, D W; Paz-Y-Miño, A; Solís, M B; Arias, L M

    2017-06-01

    There is a dearth of data from Ecuador on the burden of life-threatening fungal disease entities; therefore, we estimated the burden of serious fungal infections in Ecuador based on the populations at risk and available epidemiological databases and publications. A full literature search was done to identify all epidemiology papers reporting fungal infection rates. WHO, ONU-AIDS, Index Mundi, Global Asthma Report, Globocan, and national data [Instituto Nacional de Estadística y Censos (INEC), Ministerio de Salud Pública (MSP), Sociedad de Lucha Contra el Cáncer (SOLCA), Instituto Nacional de Donación y Trasplante de Órganos, Tejidos y Células (INDOT)] were reviewed. When no data existed, risk populations were used to estimate frequencies of fungal infections, using previously described methodology by LIFE. Ecuador has a variety of climates from the cold of the Andes through temperate to humid hot weather at the coast and in the Amazon basin. Ecuador has a population of 15,223,680 people and an average life expectancy of 76 years. The median estimate of the human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) population at risk for fungal disease (Ecuador is affected by serious fungal infection.

  13. Structural Analysis of Fungal Cerebrosides

    Directory of Open Access Journals (Sweden)

    Eliana eBarreto-Bergter

    2011-12-01

    Full Text Available Of the ceramide monohexosides (CMHs, gluco- and galactosylceramides are the main neutral glycosphingolipids expressed in fungal cells. Their structural determination is greatly dependent on the use of mass spectrometric techniques, including fast atom bombardment-mass spectrometry (FAB-MS, electrospray ionization (ESI-MS, and energy collision-induced dissociation mass spectrometry (ESI-MS/CID-MS. Nuclear magnetic resonance (NMR has also been used successfully. Such a combination of techniques, combined with classical analytical separation, such as HPTLC and column chromatography, has led to the structural elucidation of a great number of fungal CMHs. The structure of fungal CMH is conserved among fungal species and consists of a glucose or galactose residue attached to a ceramide moiety containing 9-methyl-4,8-sphingadienine with an amidic linkage to hydroxylated fatty acids, most commonly having 16 or 18 carbon atoms and unsaturation between C-3 and C-4. Along with their unique structural characteristics, fungal CMHs have a peculiar subcellular distribution and striking biological properties. Fungal cerebrosides were also characterized as antigenic molecules directly or indirectly involved in cell growth or differentiation in Schizophyllum commune, Cryptococcus neoformans, Pseudallescheria boydii, Candida albicans, Aspergillus nidulans, A.fumigatus and Colletotrichum gloeosporioides. Besides classical techniques for cerebroside (CMH analysis, we now describe new approaches, combining conventional TLC and mass spectrometry, as well as emerging technologies for subcellular localization and distribution of glycosphingolipids by SIMS and imaging MALDI TOF .

  14. Rational reprogramming of fungal polyketide first-ring cyclization.

    Science.gov (United States)

    Xu, Yuquan; Zhou, Tong; Zhou, Zhengfu; Su, Shiyou; Roberts, Sue A; Montfort, William R; Zeng, Jia; Chen, Ming; Zhang, Wei; Lin, Min; Zhan, Jixun; Molnár, István

    2013-04-02

    Resorcylic acid lactones and dihydroxyphenylacetic acid lactones represent important pharmacophores with heat shock response and immune system modulatory activities. The biosynthesis of these fungal polyketides involves a pair of collaborating iterative polyketide synthases (iPKSs): a highly reducing iPKS with product that is further elaborated by a nonreducing iPKS (nrPKS) to yield a 1,3-benzenediol moiety bridged by a macrolactone. Biosynthesis of unreduced polyketides requires the sequestration and programmed cyclization of highly reactive poly-β-ketoacyl intermediates to channel these uncommitted, pluripotent substrates to defined subsets of the polyketide structural space. Catalyzed by product template (PT) domains of the fungal nrPKSs and discrete aromatase/cyclase enzymes in bacteria, regiospecific first-ring aldol cyclizations result in characteristically different polyketide folding modes. However, a few fungal polyketides, including the dihydroxyphenylacetic acid lactone dehydrocurvularin, derive from a folding event that is analogous to the bacterial folding mode. The structural basis of such a drastic difference in the way a PT domain acts has not been investigated until now. We report here that the fungal vs. bacterial folding mode difference is portable on creating hybrid enzymes, and we structurally characterize the resulting unnatural products. Using structure-guided active site engineering, we unravel structural contributions to regiospecific aldol condensations and show that reshaping the cyclization chamber of a PT domain by only three selected point mutations is sufficient to reprogram the dehydrocurvularin nrPKS to produce polyketides with a fungal fold. Such rational control of first-ring cyclizations will facilitate efforts to the engineered biosynthesis of novel chemical diversity from natural unreduced polyketides.

  15. Cellulolytic enzyme compositions and uses thereof

    Science.gov (United States)

    Iyer, Prashant; Gaspar, Armindo Ribiero; Croonenberghs, James; Binder, Thomas P.

    2017-07-25

    The present invention relates enzyme composition comprising a cellulolytic preparation and an acetylxylan esterase (AXE); and the used of cellulolytic enzyme compositions for hydrolyzing acetylated cellulosic material. Finally the invention also relates to processes of producing fermentation products from acetylated cellulosic materials using a cellulolytic enzyme composition of the invention.

  16. Immobilization of Enzymes in Polymer Supports.

    Science.gov (United States)

    Conlon, Hugh D.; Walt, David R.

    1986-01-01

    Two experiments in which an enzyme is immobilized onto a polymeric support are described. The experiments (which also demonstrate two different polymer preparations) involve: (1) entrapping an enzyme in an acrylamide polymer; and (2) reacting the amino groups on the enzyme's (esterase) lysine residues with an activated polymer. (JN)

  17. Fungal Infections in Some Economically Important Freshwater Fishes

    Directory of Open Access Journals (Sweden)

    Zafar Iqbal*, Uzma Sheikh and Rabia Mughal

    2012-06-01

    Full Text Available Aim of this study was to investigate fungal infections in four species of carps including goldfish, Carassius (C. auratus L.; silver carp, Hypophthalmichthys (H. molitrix Richardsons; rahu, Labeo (L. rohita Hamilton and Ctenopharyngodon (C. idella Valenciennes. Nine specimens of each species were studied for the presence of fungal infections. Infected fishes showed clinical signs such as fungal growth on skin, fins, eyes, eroded fins and scales, hemorrhages on body surface and abdominal distension. The specimens from infected organs of fish were inoculated on each, malt extract, Sabouraud dextrose and potato dextrose agars. The fungal colonies of white, black, green, grey and brown colors were observed in the agar plates. Slides were prepared and stained with 0.05% Trypan blue in lactophenol. C. auratus showed the highest infection rate (44.4% followed by H. molitrix and L. rohita (11.1% each. Five fungal species viz. Aspergillus (33.3%, Penicillium (22.2%, Alternaria (27.7%, Blastomyces spp (11.1% and Rhizopus (5.5% were isolated. Posterior part of the fish had significantly (P=0.05 higher (62.5% infection as compared to anterior part (37.5%. The caudal fin with 31.25% infection was the single most affected area. This study showed that most of the fungi isolated from fishes are considered as normal mycoflora, yet many fungi can cause natural infections in ponds and aquarium.

  18. Lignin depolymerization by fungal secretomes and a microbial sink

    Energy Technology Data Exchange (ETDEWEB)

    Salvachúa, Davinia; Katahira, Rui; Cleveland, Nicholas S.; Khanna, Payal; Resch, Michael G.; Black, Brenna A.; Purvine, Samuel O.; Zink, Erika M.; Prieto, Alicia; Martínez, María J.; Martínez, Angel T.; Simmons, Blake A.; Gladden, John M.; Beckham, Gregg T.

    2016-08-25

    In Nature, powerful oxidative enzymes secreted by white rot fungi and some bacteria catalyze lignin depolymerization and some microbes are able to catabolize the resulting aromatic compounds as carbon and energy sources. Taken together, these two processes offer a potential route for microbial valorization of lignin. However, many challenges remain in realizing this concept, including that oxidative enzymes responsible for lignin depolymerization also catalyze polymerization of low molecular weight (LMW) lignin. Here, multiple basidiomycete secretomes were screened for ligninolytic enzyme activities in the presence of a residual lignin solid stream from a corn stover biorefinery, dubbed DMR-EH (Deacetylation, Mechanical Refining, and Enzymatic Hydrolysis) lignin. Two selected fungal secretomes, with high levels of laccases and peroxidases, were utilized for DMR-EH lignin depolymerization assays. The secretome from Pleurotus eryngii, which exhibited the highest laccase activity, reduced the lignin average molecular weight by 63% and 75% at pH 7 compared to the Mw of the control treated at the same conditions and the initial DMR-EH lignin, respectively, and was applied in further depolymerization assays as a function of time. As repolymerization was observed after 3 days of incubation, an aromatic-catabolic microbe (Pseudomonas putida KT2440) was incubated with the fungal secretome and DMR-EH lignin. These experiments demonstrated that the presence of the bacterium enhances lignin depolymerization, likely due to bacterial catabolism of LMW lignin, which may partially prevent repolymerization. In addition, proteomics was also applied to the P. eryngii secretome to identify the enzymes present in the fungal cocktail utilized for the depolymerization assays, which highlighted a significant number of glucose/ methanol/choline (GMC) oxidoreductases and laccases. Overall, this study demonstrates that ligninolytic enzymes can be used to partially depolymerize a solid, high

  19. Fungal treated lignocellulosic biomass as ruminant feed ingredient: a review.

    Science.gov (United States)

    van Kuijk, S J A; Sonnenberg, A S M; Baars, J J P; Hendriks, W H; Cone, J W

    2015-01-01

    In ruminant nutrition, there is an increasing interest for ingredients that do not compete with human nutrition. Ruminants are specialists in digesting carbohydrates in plant cell walls; therefore lignocellulosic biomass has potential in ruminant nutrition. The presence of lignin in biomass, however, limits the effective utilization of cellulose and hemicellulose. Currently, most often chemical and/or physical treatments are used to degrade lignin. White rot fungi are selective lignin degraders and can be a potential alternative to current methods which involve potentially toxic chemicals and expensive equipment. This review provides an overview of research conducted to date on fungal pretreatment of lignocellulosic biomass for ruminant feeds. White rot fungi colonize lignocellulosic biomass, and during colonization produce enzymes, radicals and other small compounds to breakdown lignin. The mechanisms on how these fungi degrade lignin are not fully understood, but fungal strain, the origin of lignocellulose and culture conditions have a major effect on the process. Ceriporiopsis subvermispora and Pleurotus eryngii are the most effective fungi to improve the nutritional value of biomass for ruminant nutrition. However, conclusions on the effectiveness of fungal delignification are difficult to draw due to a lack of standardized culture conditions and information on fungal strains used. Methods of analysis between studies are not uniform for both chemical analysis and in vitro degradation measurements. In vivo studies are limited in number and mostly describing digestibility after mushroom production, when the fungus has degraded cellulose to derive energy for fruit body development. Optimization of fungal pretreatment is required to shorten the process of delignification and make it more selective for lignin. In this respect, future research should focus on optimization of culture conditions and gene expression to obtain a better understanding of the mechanisms

  20. Hydrolytic enzymes in the central vacuole of plant cells.

    Science.gov (United States)

    Boller, T; Kende, H

    1979-06-01

    The hydrolase content of vacuoles isolated from protoplasts of suspension-cultured tobacco cells, of tulip petals, and of pineapple leaves, and the sedimentation behavior of tobacco tonoplasts were studied. Three precautions were found to be important for the analysis of vacuolar hydrolases and of the tonoplast. (a) Purification of protoplasts in a Ficoll gradient was necessary to remove cell debris which contained contaminating hydrolases adsorbed from the fungal cell-wall-degrading enzyme preparation. (b) Hydrolase activities in the homogenates of the intact cells or the tissue used and of the purified protoplasts had to be compared to verify the absence of contaminating hydrolases in the protoplast preparation. (c) Vacuoles obtained from the protoplasts by an osmotic shock had to be purified from the lysate in a Ficoll gradient. Since the density of the central vacuole approximates that of the protoplasts, about a 10% contamination of the vacuolar preparation by surviving protoplasts could not be eliminated and had to be taken into account when the distribution of enzymes and of radioactivity was calculated.THE INTRACELLULAR ACTIVITIES OF THE FOLLOWING ACID HYDROLASES WERE PRIMARILY LOCALIZED IN THE VACUOLE OF TOBACCO CELLS: alpha-mannosidase, beta-N-acetylglucosaminidase, beta-fructosidase, nuclease, phosphatase, phosphodiesterase. A similar composition of acid hydrolases was found in vacuoles obtained from protoplasts of tulip petals. Proteinase, a hydrolase with low activity in tobacco cells and tulip petals and therefore difficult to localize unequivocally, was found to be vacuolar in pineapple leaves, a tissue containing high levels of this enzyme. Our data support the hypothesis that the central vacuole of higher plant cells has an enzyme composition analogous to that of the animal lysosome.None of the vacuolar enzymes investigated was found to be bound to the tonoplast. When vacuoles were isolated from cells labeled with radioactive choline, the vacuolar

  1. Effective pH pretreatment and cell disruption method for real-time intracellular enzyme activity assay of a marine fungus covered with pigments.

    Science.gov (United States)

    Zhang, Xiaoxu; Gao, Yanyun; Yin, Ying; Cai, Menghao; Zhou, Xiangshan; Zhang, Yuanxing

    2017-02-07

    Filamentous fungi are capable producers of many bioactive compounds, and real-time intracellular enzyme activity assay is an essential guidance for their bioprocess developments. However, there are many difficulties in preparing homogenate for enzyme activity assay, such as disrupting fungal cell with complicated cellular structure and solid cell wall, removing abundant extracellular metabolites accumulating on mycelia, and so on. Halorosellinia sp. (No. 1403) was a marine-derived filamentous fungus producing a potential antitumor compound 1403C, and the deep red pigments (with main component of 1403C) covering on its mycelia showed strong absorption in a wide range, which critically affected the measurement of many enzyme activities. In this study, we developed an effective pH pretreatment and cell disruption method to prepare homogenate for enzyme activity assay. When mycelia were washed by the solution with pH 5.0 for 3 min, most pigments could be removed without severe loss on enzyme activities. Afterward, grinding with mini bead for 15 min with alternating cooling could effectively disrupt both cell wall and mitochondrial membrane. These methods have been successfully applied on real-time intracellular enzyme activity assay of Halorosellinia sp. (No. 1403) and can offer enlightenment for other filamentous fungi with similar problems.

  2. Enzyme immunoassay

    DEFF Research Database (Denmark)

    Feldt-Rasmussen, B; Dinesen, B; Deckert, M

    1985-01-01

    An enzyme linked immunoadsorbent assay for urinary albumin using commercially available reagents is described. The assay range is 2.5-120 micrograms/l. When samples are analysed in two standard dilutions, the assayable albumin concentration range is 2.5-240 mg/l, covering the clinical range from...

  3. Serious fungal infections in Chile.

    Science.gov (United States)

    Alvarez Duarte, E; Denning, D W

    2017-06-01

    The incidence and prevalence of fungal infections in Chile are unknown. Here, we have estimated the burden of serious fungal diseases from data obtained from clinical reports, WHO reports, Chilean census, OECD reports and comprehensive literature search available on PubMed and SciELO, among other scientific resources. Due the lack of official data about fungal diseases, frequencies were calculated based on the specific populations at risk. Recurrent vulvovaginal candidiasis (>4 episodes/year) is estimated to occur in 3108/100,000. Using a low international average rate of 5/100,000, we estimate 878 candidaemia cases and 132 patients with intra-abdominal candidiasis. Due to the low incidence of pulmonary tuberculosis (TB) in Chile, limited numbers of patients with chronic pulmonary aspergillosis are likely: a total of 1212, 25% following TB. Invasive aspergillosis is estimated to affect 296 patients following leukaemia therapy, transplantation and chronic obstructive pulmonary disease (COPD), 1.7/100,000. In addition, allergic bronchopulmonary aspergillosis (ABPA) and severe asthma with fungal sensitisation (SAFS) were estimated to be around 97.9/100,000 and 127/100,000 respectively, in 675,772 adult asthmatics and 1700 CF patients. Given a 38,000 human immunodeficiency virus (HIV) population, with around 2189 new cases of acquired immune deficiency syndrome (AIDS) annually, cryptococcal meningitis and Pneumocystis pneumonia are estimated at 0.12/100,000 and 4.3/100,000, respectively. In total, 325,000 (1.9%) people in Chile develop serious fungal infections annually. Respiratory fungal disease predominates in Chile; a national action plan for fungal disease is urgently needed, including epidemiological studies to validate the estimates.

  4. Evaluation of pulmonary fungal diseases in patients with fungal rhino-sinusitis

    Directory of Open Access Journals (Sweden)

    M.Sh. Badawy

    2013-07-01

    Conclusion: Universal screening for pulmonary fungal infection especially in patients with fungal rhino sinusitis is highly recommended to treat it early, decrease morbidity and mortality of the diseases.

  5. Fungal sensing of host environment.

    Science.gov (United States)

    Braunsdorf, C; Mailänder-Sánchez, D; Schaller, M

    2016-09-01

    To survive inside a host, fungi have to adapt to a changing and often hostile environment and therefore need the ability to recognize what is going on around them. To adapt to different host niches, they need to sense external conditions such as temperature, pH and to recognize specific host factors. The ability to respond to physiological changes inside the host, independent of being in a commensal, pathogenic or even symbiotic context, implicates mechanisms for sensing of specific host factors. Because the cell wall is constantly in contact with the surrounding, fungi express receptors on the surface of their cell wall, such as pheromone receptors, which have important roles, besides mediating chemotropism for mating. We are not restricting the discussion to the human host because the receptors and mechanisms used by different fungal species to sense their environment are often similar even for plant pathogens. Furthermore, the natural habitat of opportunistic pathogenic fungi with the potential to cause infection in a human host is in soil and on plants. While the hosts' mechanisms of sensing fungal pathogens have been addressed in the literature, the focus of this review is to fill the gap, giving an overview on fungal sensing of a host-(ile) environment. Expanding our knowledge on host-fungal interactions is extremely important to prevent and treat diseases of pathogenic fungi, which are important issues in human health and agriculture but also to understand the delicate balance of fungal symbionts in our ecosystem. © 2016 John Wiley & Sons Ltd.

  6. Microbiological diagnostics of fungal infections

    Directory of Open Access Journals (Sweden)

    Corrado Girmenia

    2013-07-01

    Full Text Available Laboratory tests for the detection of fungal infections are easy to perform. The main obstacle to a correct diagnosis is the correlation between the laboratory findings and the clinical diagnosis. Among pediatric patients, the most common fungal pathogen is Candida. The detection of fungal colonization may be performed through the use of chromogenic culture media, which allows also the identification of Candida subspecies, from which pathogenicity depends. In neonatology, thistest often drives the decision to begin a empiric therapy; in this regard, a close cooperation between microbiologists and clinicians is highly recommended. Blood culture, if positive, is a strong confirmation of fungal infection; however, its low sensitivity results in a high percentage of false negatives, thus decreasing its reliability. Molecular diagnostics is still under evaluation, whereas the detection of some fungal antigens, such as β-D-glucan, galactomannan, mannoprotein, and cryptococcal antigen in the serum is used for adults, but still under evaluations for pediatric patients.http://dx.doi.org/10.7175/rhc.v4i1S.862

  7. Fungal laccases: versatile tools for lignocellulose transformation.

    Science.gov (United States)

    Piscitelli, Alessandra; Del Vecchio, Claudia; Faraco, Vincenza; Giardina, Paola; Macellaro, Gemma; Miele, Annalisa; Pezzella, Cinzia; Sannia, Giovanni

    2011-11-01

    Conversion of lignocellulosic materials to useful, high value products normally requires a pre-treatment step to transform or deconstruct the recalcitrant and heterogeneous lignin fraction. The development of "green tools" for the transformation of lignocellulosic feedstocks is in high demand for a sustainable exploitation of such resources. This multi-faceted challenge is being addressed by an ever-increasing suite of ligninolytic enzymes isolated from various sources. Among these, fungal laccases are known to play an important role in lignin degradation/modification processes. The white-rot fungus Pleurotus ostreatus expresses multiple laccase genes encoding isoenzymes with different properties. The availability of established recombinant expression systems for P. ostreatus laccase isoenzymes has allowed to further enrich the panel of P. ostreatus laccases by the construction of mutated, "better performing" enzymes through molecular evolution techniques. New oxidative catalysts with improved activity and stability either at high temperature and at acidic and alkaline pH have been isolated and characterized. Copyright © 2011 Académie des sciences. Published by Elsevier SAS. All rights reserved.

  8. Bacterial and fungal markers in tobacco smoke

    Energy Technology Data Exchange (ETDEWEB)

    Szponar, B., E-mail: szponar@iitd.pan.wroc.pl [Lund University, Dept. of Laboratory Medicine, Soelvegatan 23, 223 62 Lund (Sweden); Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla 12, 53-114 Wroclaw (Poland); Pehrson, C.; Larsson, L. [Lund University, Dept. of Laboratory Medicine, Soelvegatan 23, 223 62 Lund (Sweden)

    2012-11-01

    Previous research has demonstrated that cigarette smoke contains bacterial and fungal components including lipopolysaccharide (LPS) and ergosterol. In the present study we used gas chromatography-mass spectrometry to analyze tobacco as well as mainstream and second hand smoke for 3-hydroxy fatty acids (3-OH FAs) of 10 to 18 carbon chain lengths, used as LPS markers, and ergosterol, used as a marker of fungal biomass. The air concentrations of LPS were 0.0017 nmol/m{sup 3} (N = 5) and 0.0007/m{sup 3} (N = 6) in the smoking vs. non-smoking rooms (p = 0.0559) of the studied private houses, and 0.0231 nmol/m{sup 3} (N = 5) vs. 0.0006 nmol/m{sup 3} (N = 5) (p = 0.0173), respectively, at the worksite. The air concentrations of ergosterol were also significantly higher in rooms with ongoing smoking than in rooms without smoking. A positive correlation was found between LPS and ergosterol in rooms with smoking but not in rooms without smoking. 3-OH C14:0 was the main 3-OH FA, followed by 3-OH C12:0, both in mainstream and second hand smoke and in phenol:water smoke extracts prepared in order to purify the LPS. The Limulus activity of the phenolic phase of tobacco was 3900 endotoxin units (EU)/cigarette; the corresponding amount of the smoke, collected on filters from 8 puffs, was 4 EU/cigarette. Tobacco smoking has been associated with a range of inflammatory airway conditions including COPD, asthma, bronchitis, alveolar hypersensitivity etc. Significant levels of LPS and ergosterol were identified in tobacco smoke and these observations support the hypothesis that microbial components of tobacco smoke contribute to inflammation and airway disease. -- Highlights: Black-Right-Pointing-Pointer Air concentration of bacterial and fungal markers is significantly higher in rooms with ongoing smoking than without smoking. Black-Right-Pointing-Pointer Bacterial LPS correlates with fungal marker in rooms with ongoing smoking but not without smoking. Black-Right-Pointing-Pointer LPS

  9. Functional characterization of salicylate hydroxylase from the fungal endophyte Epichloë festucae.

    Science.gov (United States)

    Ambrose, Karen V; Tian, Zipeng; Wang, Yifei; Smith, Jordan; Zylstra, Gerben; Huang, Bingru; Belanger, Faith C

    2015-06-09

    Epichloë spp. are symbiotic fungal endophytes of many cool season grasses. The presence of the fungal endophytes often confers insect, drought, and disease tolerance to the host grasses. The presence of the fungal endophytes within the host plants does not elicit host defense responses. The molecular basis for this phenomenon is not known. Epichloë festucae, the endophyte of Festuca rubra, expresses a salicylate hydroxylase similar to NahG from the bacterium Pseudomonas putida. Few fungal salicylate hydroxylase enzymes have been reported. The in planta expression of an endophyte salicylate hydroxylase raised the possibility that degradation of plant-produced salicylic acid is a factor in the mechanism of how the endophyte avoids eliciting host plant defenses. Here we report the characterization of the E. festucae salicylate hydroxylase, designated Efe-shyA. Although the fungal enzyme has the expected activity, based on salicylic acid levels in endophyte-free and endophyte-infected plants it is unlikely that expression of the endophyte salicylate hydroxylase is a factor in the lack of a host defense response to the presence of the fungal endophyte.

  10. Functional characterization of salicylate hydroxylase from the fungal endophyte Epichloë festucae

    Science.gov (United States)

    Ambrose, Karen V.; Tian, Zipeng; Wang, Yifei; Smith, Jordan; Zylstra, Gerben; Huang, Bingru; Belanger, Faith C.

    2015-01-01

    Epichloë spp. are symbiotic fungal endophytes of many cool season grasses. The presence of the fungal endophytes often confers insect, drought, and disease tolerance to the host grasses. The presence of the fungal endophytes within the host plants does not elicit host defense responses. The molecular basis for this phenomenon is not known. Epichloë festucae, the endophyte of Festuca rubra, expresses a salicylate hydroxylase similar to NahG from the bacterium Pseudomonas putida. Few fungal salicylate hydroxylase enzymes have been reported. The in planta expression of an endophyte salicylate hydroxylase raised the possibility that degradation of plant-produced salicylic acid is a factor in the mechanism of how the endophyte avoids eliciting host plant defenses. Here we report the characterization of the E. festucae salicylate hydroxylase, designated Efe-shyA. Although the fungal enzyme has the expected activity, based on salicylic acid levels in endophyte-free and endophyte-infected plants it is unlikely that expression of the endophyte salicylate hydroxylase is a factor in the lack of a host defense response to the presence of the fungal endophyte. PMID:26055188

  11. The Fungal Defensin Family Enlarged

    Directory of Open Access Journals (Sweden)

    Jiajia Wu

    2014-08-01

    Full Text Available Fungi are an emerging source of peptide antibiotics. With the availability of a large number of model fungal genome sequences, we can expect that more and more fungal defensin-like peptides (fDLPs will be discovered by sequence similarity search. Here, we report a total of 69 new fDLPs encoded by 63 genes, in which a group of fDLPs derived from dermatophytes are defined as a new family (fDEF8 according to sequence and phylogenetic analyses. In the oleaginous fungus Mortierella alpine, fDLPs have undergone extensive gene expansion. Our work further enlarges the fungal defensin family and will help characterize new peptide antibiotics with therapeutic potential.

  12. Enzyme Mimics: Advances and Applications.

    Science.gov (United States)

    Kuah, Evelyn; Toh, Seraphina; Yee, Jessica; Ma, Qian; Gao, Zhiqiang

    2016-06-13

    Enzyme mimics or artificial enzymes are a class of catalysts that have been actively pursued for decades and have heralded much interest as potentially viable alternatives to natural enzymes. Aside from having catalytic activities similar to their natural counterparts, enzyme mimics have the desired advantages of tunable structures and catalytic efficiencies, excellent tolerance to experimental conditions, lower cost, and purely synthetic routes to their preparation. Although still in the midst of development, impressive advances have already been made. Enzyme mimics have shown immense potential in the catalysis of a wide range of chemical and biological reactions, the development of chemical and biological sensing and anti-biofouling systems, and the production of pharmaceuticals and clean fuels. This Review concerns the development of various types of enzyme mimics, namely polymeric and dendrimeric, supramolecular, nanoparticulate and proteinic enzyme mimics, with an emphasis on their synthesis, catalytic properties and technical applications. It provides an introduction to enzyme mimics and a comprehensive summary of the advances and current standings of their applications, and seeks to inspire researchers to perfect the design and synthesis of enzyme mimics and to tailor their functionality for a much wider range of applications. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Enzyme-carrying electrospun nanofibers.

    Science.gov (United States)

    Jia, Hongfei

    2011-01-01

    Compared to other nanomaterials as supports for enzyme immobilization, nanofibers provide a promising configuration in balancing the key factors governing the catalytic performance of the immobilized enzymes including surface area-to-volume ratio, mass transfer resistance, effective loading, and the easiness to recycle. Synthetic and natural polymers can be fabricated into nanofibers via a physical process called electrospinning. The process requires only simple apparatus to operate, yet has proved to be very flexible in the selection of feedstock materials and also effective to control and manipulate the properties of the resulting nanofibers such as size and surface morphology, which are typically important parameters for enzyme immobilization supports. This chapter describes a protocol for the preparation of nanofibrous enzyme, involving the synthesis and end-group functionalization of polystyrene, production of electrospun nanofibers, and surface immobilization of enzyme via covalent attachment.

  14. Nonculture Diagnostics in Fungal Disease.

    Science.gov (United States)

    Powers-Fletcher, Margaret V; Hanson, Kimberly E

    2016-03-01

    Fungal diagnostics that utilize antibody, antigen or nucleic acid detection offer several advantages that supplement traditional culture-based methods. As a group, nonculture assays can help identify patients with invasive fungal infection (IFI) sooner than is possible with culture, are often more sensitive, and can be used to guide early interventions. Challenges associated with these techniques include the possibility for contamination or cross-reactivity as well as the potential for false negative tests. This review summarized the test characteristics and clinical utility of nonculture-based laboratory methods. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Fungal genomics beyond Saccharomyces cerevisiae?

    DEFF Research Database (Denmark)

    Hofmann, Gerald; Mcintyre, Mhairi; Nielsen, Jens

    2003-01-01

    Fungi are used extensively in both fundamental research and industrial applications. Saccharomyces cerevisiae has been the model organism for fungal research for many years, particularly in functional genomics. However, considering the diversity within the fungal kingdom, it is obvious...... that the application of the existing methods of genome, transcriptome, proteome and metabolome analysis to other fungi has enormous potential, especially for the production of food and food ingredients. The developments in the past year demonstrate that we have only just started to exploit this potential....

  16. Rapid methods for the extraction and archiving of molecular grade fungal genomic DNA.

    Science.gov (United States)

    Borman, Andrew M; Palmer, Michael; Johnson, Elizabeth M

    2013-01-01

    The rapid and inexpensive extraction of fungal genomic DNA that is of sufficient quality for molecular approaches is central to the molecular identification, epidemiological analysis, taxonomy, and strain typing of pathogenic fungi. Although many commercially available and in-house extraction procedures do eliminate the majority of contaminants that commonly inhibit molecular approaches, the inherent difficulties in breaking fungal cell walls lead to protocols that are labor intensive and that routinely take several hours to complete. Here we describe several methods that we have developed in our laboratory that allow the extremely rapid and inexpensive preparation of fungal genomic DNA.

  17. Fungal chitinases: diversity, mechanistic properties and biotechnological potential.

    Science.gov (United States)

    Hartl, Lukas; Zach, Simone; Seidl-Seiboth, Verena

    2012-01-01

    Chitin derivatives, chitosan and substituted chito-oligosaccharides have a wide spectrum of applications ranging from medicine to cosmetics and dietary supplements. With advancing knowledge about the substrate-binding properties of chitinases, enzyme-based production of these biotechnologically relevant sugars from biological resources is becoming increasingly interesting. Fungi have high numbers of glycoside hydrolase family 18 chitinases with different substrate-binding site architectures. As presented in this review, the large diversity of fungal chitinases is an interesting starting point for protein engineering. In this review, recent data about the architecture of the substrate-binding clefts of fungal chitinases, in connection with their hydrolytic and transglycolytic abilities, and the development of chitinase inhibitors are summarized. Furthermore, the biological functions of chitinases, chitin and chitosan utilization by fungi, and the effects of these aspects on biotechnological applications, including protein overexpression and autolysis during industrial processes, are discussed in this review.

  18. Organ Transplant Patients and Fungal Infections

    Science.gov (United States)

    ... Waterborne, and Environmental Diseases Mycotic Diseases Branch Organ Transplant Patients and Fungal Infections Recommend on Facebook Tweet ... Top of Page Preventing fungal infections in organ transplant patients Fungi are difficult to avoid because they ...

  19. HIV/AIDS and Fungal Infections

    Science.gov (United States)

    ... Who Gets Fungal Infections? People living with HIV/AIDS Organ Transplant Patients Cancer Patients Hospitalized Patients Stem Cell Transplant Patients Medications that Weaken Your Immune System Outbreaks Rhizopus Investigation CDC at Work Global Fungal Diseases Cryptococcal Meningitis ...

  20. Applications of Microbial Enzymes in Food Industry

    Directory of Open Access Journals (Sweden)

    Binod Parameswaran

    2018-01-01

    Full Text Available The use of enzymes or microorganisms in food preparations is an age-old process. With the advancement of technology, novel enzymes with wide range of applications and specificity have been developed and new application areas are still being explored. Microorganisms such as bacteria, yeast and fungi and their enzymes are widely used in several food preparations for improving the taste and texture and they offer huge economic benefits to industries. Microbial enzymes are the preferred source to plants or animals due to several advantages such as easy, cost-effective and consistent production. The present review discusses the recent advancement in enzyme technology for food industries. A comprehensive list of enzymes used in food processing, the microbial source of these enzymes and the wide range of their application are discussed.

  1. Biocombinatorial Engineering of Fungal PKS-NRPS Hybrids for Production of Novel Synthetic Natural Products

    DEFF Research Database (Denmark)

    Nielsen, Maria Lund

    Natural products have had a huge impact on the history of drug development, and today, these compounds continue to be a key source of novel drug leads. Over the years, natural products have been exploited for their pharmacological and biological activities in the treatment of many diseases....... Secondary metabolites (SMs) from bacteria, plants and filamentous fungi constitute a large group of important natural products. In this thesis I explore the biosynthesis of several fungal SMs along with the enzymes responsible for their synthesis. Specifically, my research focuses on the expression...... and engineering of a certain type of fungal enzymes – natural fusions of polyketide synthases and nonribosomal peptide synthetases (PKS-NRPSs). The thesis is divided into two topics: 1) Expanding fungal chemodiversity through combinatorial biosynthesis 2) Two CRISPR-Cas9-based approaches to linking SMs...

  2. Microbiology of systemic fungal infections

    Directory of Open Access Journals (Sweden)

    Chakrabarti A

    2005-01-01

    Full Text Available The increased incidence of systemic fungal infections in the past two decades has been overwhelming. Earlier, it was pathogenic dimorphic fungi, which were known to cause systemic infections. However, starting from the 1960s, opportunistic fungi started causing more number of infections, especially in the immunocompromised host. More recently, newer and less common fungal agents are being increasingly associated with infection in immunosuppressed hosts. Amongst dimorphic fungi, infections due to Histoplasma capsulatum and Penicillium marneffei are increasingly reported in patients with AIDS in India. H. capsulatum is found country wide, but P. marneffei remains restricted to Manipur state. Although both varieties of C. neoformans , C. neoformans var. neoformans (serotypes A & D, and C. neoformans var. gattii (serotypes B & C are reported in India, most of the cases reported are of serotype A. Increased incidence of cryptococcosis is reported from all centers with the emergence of AIDS. Systemic infection due to species under Candida , Aspergillus and zygomycetes is widely prevalent in nosocomial setting, and outbreaks due to unusual fungi are reported occasionally from tertiary care centers. This global change in systemic fungal infections has emphasized the need to develop good diagnostic mycology laboratories in this country and to recognize this increasingly large group of potential fungal pathogens.

  3. Hidden killers: human fungal infections

    NARCIS (Netherlands)

    Brown, G.D.; Denning, D.W.; Gow, N.A.; Levitz, S.M.; Netea, M.G.; White, T.C.

    2012-01-01

    Although fungal infections contribute substantially to human morbidity and mortality, the impact of these diseases on human health is not widely appreciated. Moreover, despite the urgent need for efficient diagnostic tests and safe and effective new drugs and vaccines, research into the

  4. Fungal Endophytes: Beyond Herbivore Management

    Directory of Open Access Journals (Sweden)

    Bamisope S. Bamisile

    2018-03-01

    Full Text Available The incorporation of entomopathogenic fungi as biocontrol agents into Integrated Pest Management (IPM programs without doubt, has been highly effective. The ability of these fungal pathogens such as Beauveria bassiana and Metarhizium anisopliae to exist as endophytes in plants and protect their colonized host plants against the primary herbivore pests has widely been reported. Aside this sole role of pest management that has been traditionally ascribed to fungal endophytes, recent findings provided evidence of other possible functions as plant yield promoter, soil nutrient distributor, abiotic stress and drought tolerance enhancer in plants. However, reports on these additional important effects of fungal endophytes on the colonized plants remain scanty. In this review, we discussed the various beneficial effects of endophytic fungi on the host plants and their primary herbivore pests; as well as some negative effects that are relatively unknown. We also highlighted the prospects of our findings in further increasing the acceptance of fungal endophytes as an integral part of pest management programs for optimized crop production.

  5. Fungal Endophytes: Beyond Herbivore Management

    Science.gov (United States)

    Bamisile, Bamisope S.; Dash, Chandra K.; Akutse, Komivi S.; Keppanan, Ravindran; Wang, Liande

    2018-01-01

    The incorporation of entomopathogenic fungi as biocontrol agents into Integrated Pest Management (IPM) programs without doubt, has been highly effective. The ability of these fungal pathogens such as Beauveria bassiana and Metarhizium anisopliae to exist as endophytes in plants and protect their colonized host plants against the primary herbivore pests has widely been reported. Aside this sole role of pest management that has been traditionally ascribed to fungal endophytes, recent findings provided evidence of other possible functions as plant yield promoter, soil nutrient distributor, abiotic stress and drought tolerance enhancer in plants. However, reports on these additional important effects of fungal endophytes on the colonized plants remain scanty. In this review, we discussed the various beneficial effects of endophytic fungi on the host plants and their primary herbivore pests; as well as some negative effects that are relatively unknown. We also highlighted the prospects of our findings in further increasing the acceptance of fungal endophytes as an integral part of pest management programs for optimized crop production.

  6. Imaging fungal infections in children

    NARCIS (Netherlands)

    Ankrah, Alfred O.; Sathekge, Mike M; Dierckx, Rudi A.J.O.; Glaudemans, Andor W.J.M.

    Fungal infections in children rarely occur, but continue to have a high morbidity and mortality despite the development of newer antifungal agents. It is essential for these infections to be diagnosed at the earliest possible stage so appropriate treatment can be initiated promptly. The addition of

  7. [Fungal infections of the gastrointestinal tract].

    Science.gov (United States)

    Maragkoudakis, Emmanouil; Realdi, Giuseppe; Dore, Maria Pina

    2005-06-01

    In immunocompetent subjects fungal infections of the gastrointestinal tract are uncommon. Candida esophagitis remains the single most common fungal infection in immunocompromised hosts or in H. pylori- infected patients who receive antibiotic therapy. Enteric fungal infections are uncommon even in HIV-infected patients. Antifungal agents such as amphotericin B, ketoconazole, fluconazole, and the various formulations of itraconazole are effective for most cases.

  8. Has substrate-dependent co-evolution of enzyme function occured in the attine ant-fungus symbiosis

    DEFF Research Database (Denmark)

    de Fine Licht, Henrik Hjarvard; Schiøtt, Morten; Boomsma, Jacobus Jan

    as substrate for their fungus gardens, whereas the more basal attine genera use substrates such as flowers, plant debris, small twigs, insect feces and insect carcasses. This diverse array of fungal substrates across the attine lineage implies that the symbiotic fungus needs different enzymes to break down...... the plant material that the ants provide or different efficiencies of enzyme function. Here we present the fist partial amino acid sequences from a fungal xylanase gene to test the hypothesis that fungal enzymes that degrade plant cell walls have functionally co-evolved with the ants....

  9. The role of enzymes in fungus-growing ant evolution

    DEFF Research Database (Denmark)

    de Fine Licht, Henrik Hjarvard

    behaviour. Here we report the first large-scale comparative study on fungus garden enzyme profiles and show that various interesting changes can be documented. A more detailed analysis of laccase expression, an enzyme that is believed to oxidize phenols in defensive secondary plant compounds such as tannins......, showed that this enzyme is exclusively found in the gardens of leaf-cutting ants, where it is significantly upregulated in the gongylidia. I’ll discuss the possible role of this enzyme and other fungal modifications in the evolution of the leafcutter ants and their non-leafcutting attine relatives....

  10. Clarification of orange juice by crude fungal pectinase from citrus peel

    African Journals Online (AJOL)

    Fungal pectinase enzyme was produced by Rhizopus oryzae on a solid culture containing citrus peel of orange (35% w/v). The crude extract with maximum pectinase activity of 1, 360 u/ml was used to clarify orange juice. The yield, turbidity and viscosity as well as pH, total soluble solids, ascorbic acids and total titratable ...

  11. A rapid lateral flow immunoassay for the detection of fungal alpha-amylase at the workplace

    NARCIS (Netherlands)

    Koets, M.; Sander, I.; Bogdanovic, J.; Doekes, G.; Amerongen, van A.

    2006-01-01

    Fungal alpha-amylase is a flour supplement which is added to improve the quality of bakery products. Various studies have shown that exposure to this enzyme is an important risk factor for the development of bakers allergy and this allergy is reported to be one of the most frequent causes of

  12. Fungal secretomics to probe the biological functions of lytic polysaccharide monooxygenases

    DEFF Research Database (Denmark)

    Berrin, Jean-Guy; Rosso, Marie-Noëlle; Abou Hachem, Maher

    2017-01-01

    to starch, the main carbon storage reservoir. In this review, we focus on the identification of lytic polysaccharide monooxygenases (LPMOs) and their redox partners in fungal secretomes to highlight the biological functions of these remarkable enzyme systems and we discuss future trends related to LPMO...

  13. Subseafloor basalts as fungal habitats

    Directory of Open Access Journals (Sweden)

    M. Ivarsson

    2012-09-01

    Full Text Available The oceanic crust is believed to host the largest potential habitat for microbial life on Earth, yet, still we lack substantial information about the abundance, diversity, and consequence of its biosphere. The last two decades have involved major research accomplishments within this field and a change in view of the ocean crust and its potential to harbour life. Here fossilised fungal colonies in subseafloor basalts are reported from three different seamounts in the Pacific Ocean. The fungal colonies consist of various characteristic structures interpreted as fungal hyphae, fruit bodies and spores. The fungal hyphae are well preserved with morphological characteristics such as hyphal walls, septa, thallic conidiogenesis, and hyphal tips with hyphal vesicles within. The fruit bodies consist of large (∼50–200 µm in diameter body-like structures with a defined outer membrane and an interior filled with calcite. The fruit bodies have at some stage been emptied of their contents of spores and filled by carbonate-forming fluids. A few fruit bodies not filled by calcite and with spores still within support this interpretation. Spore-like structures (ranging from a few µm to ∼20 µm in diameter are also observed outside of the fruit bodies and in some cases concentrated to openings in the membrane of the fruit bodies. The hyphae, fruit bodies and spores are all closely associated with a crust lining the vein walls that probably represent a mineralized biofilm. The results support a fungal presence in deep subseafloor basalts and indicate that such habitats were vital between ∼81 and 48 Ma.

  14. Immune Recognition of Fungal Polysaccharides

    Directory of Open Access Journals (Sweden)

    Brendan D. Snarr

    2017-08-01

    Full Text Available The incidence of fungal infections has dramatically increased in recent years, in large part due to increased use of immunosuppressive medications, as well as aggressive medical and surgical interventions that compromise natural skin and mucosal barriers. There are relatively few currently licensed antifungal drugs, and rising resistance to these agents has led to interest in the development of novel preventative and therapeutic strategies targeting these devastating infections. One approach to combat fungal infections is to augment the host immune response towards these organisms. The polysaccharide-rich cell wall is the initial point of contact between fungi and the host immune system, and therefore, represents an important target for immunotherapeutic approaches. This review highlights the advances made in our understanding of the mechanisms by which the immune system recognizes and interacts with exopolysaccharides produced by four of the most common fungal pathogens: Aspergillus fumigatus, Candida albicans, Cryptococcus neoformans, and Histoplasma capsulatum. Work to date suggests that inner cell wall polysaccharides that play an important structural role are the most conserved across diverse members of the fungal kingdom, and elicit the strongest innate immune responses. The immune system senses these carbohydrates through receptors, such as lectins and complement proteins. In contrast, a greater diversity of polysaccharides is found within the outer cell walls of pathogenic fungi. These glycans play an important role in immune evasion, and can even induce anti-inflammatory host responses. Further study of the complex interactions between the host immune system and the fungal polysaccharides will be necessary to develop more effective therapeutic strategies, as well as to explore the use of immunosuppressive polysaccharides as therapeutic agents to modulate inflammation.

  15. DIAGNOSIS & MANAGEMENT OF ALLERGIC FUNGAL SINUSITIS

    Directory of Open Access Journals (Sweden)

    Syam Manohar Gadhamsetty

    2016-08-01

    Full Text Available BACKGROUND Chronic sinusitis is one of the common diagnosis in ENT practice. Allergic fungal sinusitis is a clinical entity with characteristic clinical, radiographic and histopathological findings. Allergic fungal sinusitis and eosinophilic mucin rhinosinusitis can easily be misdiagnosed. AIM OF STUDY A prospective clinical study of allergic Fungal Rhinosinusitis to use diagnostic criteria to confirm the disease with Radiological, Pathological & Microbiological investigations and their management. MATERIALS & METHODS A prospective study of allergic Fungal Rhinosinusitis in 2 years from November 2011 to October 2013. Among the patients who attended the ENT OPD during this period, 21 patients with symptoms and signs suggestive of Allergic Fungal Rhinosinusitis are selected.

  16. Aquatic Plant Control Research Program. Biological Control of Hydrilla verticillata (L.f.) Royle with Lytic Enzyme-Producing Microorganisms.

    Science.gov (United States)

    1985-09-01

    layer around bacterial colonies or fungal mycelia (Figure 3). Pectinase production was enhanced by successively subculturing (a minimum of ten times...they comprised the larger group of lytic enzyme pro- ducers. Twenty fungal isolates were cellulase positive and one was pectinase positive. Only one...index values for the pectinase positive fungal isolate (224) showed no significant difference from PDB-treated controls at any time and was also excluded

  17. Development of a commercial enzymes system for lignocellulosic biomass saccharification

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Manoj

    2012-12-20

    DSM Innovation Inc., in its four year effort was able to evaluate and develop its in-house DSM fungal cellulolytic enzymes system to reach enzyme efficiency mandates set by DoE Biomass program MYPP goals. DSM enzyme cocktail is uniquely active at high temperature and acidic pH, offering many benefits and product differentiation in 2G bioethanol production. Under this project, strain and process development, ratio optimization of enzymes, protein and genetic engineering has led to multitudes of improvement in productivity and efficiency making development of a commercial enzyme system for lignocellulosic biomass saccharification viable. DSM is continuing further improvement by additional biodiversity screening, protein engineering and overexpression of enzymes to continue to further lower the cost of enzymes for saccharification of biomass.

  18. Fungal Exopolysaccharide: Production, Composition and Applications

    Directory of Open Access Journals (Sweden)

    Subhadip Mahapatra

    2013-01-01

    Full Text Available Fungal exopolysaccharides (EPSs have been recognized as high value biomacromolecules for the last two decades. These products, including pullulan, scleroglucan, and botryosphaeran, have several applications in industries, pharmaceuticals, medicine, foods etc. Although fungal EPSs are highly relevant, to date information concerning fungal biosynthesis is scarce and an extensive search for new fugal species that can produce novel EPSs is still needed. In most cases, the molecular weight variations and sugar compositions of fungal EPSs are dependent to culture medium composition and different physical conditions provided during fermentation. An inclusive and illustrative review on fungal EPS is presented here. The general outline of the present work includes fungal EPS production, their compositions and applications. An emphasis is also given to listing out different fungal strains that can produce EPSs.

  19. Explorations of fungal biosynthesis of reduced polyketides - a personal viewpoint.

    Science.gov (United States)

    Vederas, John C

    2014-10-01

    This viewpoint on biosynthesis of reduced polyketides in fungi traces evolution of the research area over more than 4 decades. It is a companion to the related articles by two personal and scientific friends with whom there has been free exchange of ideas for over 30 years. Beginning with very rudimentary knowledge about assembly of such natural products, developments using stable isotope labelling and subsequently identification of biosynthetic genes, led to understanding of the processive nature of polyketide formation. Recent expression and isolation of fungal iterative polyketide synthase enzymes has enabled more detailed exploration of the mechanisms of these fascinating molecular machines.

  20. Cellulolytic potential of thermophilic species from four fungal orders

    DEFF Research Database (Denmark)

    Busk, Peter Kamp; Lange, Lene

    2013-01-01

    Elucidation of fungal biomass degradation is important for understanding the turnover of biological materials in nature and has important implications for industrial biomass conversion. In recent years there has been an increasing interest in elucidating the biological role of thermophilic fungi....... Thermophilic fungi are the only described eukaryotes that can grow at temperatures above 45 ºC. All 16 fungi were able to grow on crystalline cellulose but their secreted enzymes showed widely different cellulolytic activities, pH optima and thermostabilities. Interestingly, in contrast to previous reports, we...

  1. Yield of ethanol from enzyme-hydrolyzed yam (Dioscorea rotundata ...

    African Journals Online (AJOL)

    Fresh whole yam tubers and cocoyam corms were separately processed into flours by washing, peeling, blanching, slicing,drying and milling. The flours were enzyme-hydrolyzed by mixing 500g of flour with 2Lof water followed by treatment with a combination of bacterial alpha amylase, limit dextrinase and fungal alpha ...

  2. Induction of defensive enzymes (isozymes) during defense against ...

    African Journals Online (AJOL)

    These results suggest that enhancing defensive enzymes' activities and inducing new isozymes may be related to mitigating pathogen-induced oxidative damage which result in the decrease of calli decay, and this implies that antioxidant defense response may be involved in the mechanisms of plant against fungal ...

  3. Marine-derived fungi: diversity of enzymes and biotechnological applications.

    Science.gov (United States)

    Bonugli-Santos, Rafaella C; Dos Santos Vasconcelos, Maria R; Passarini, Michel R Z; Vieira, Gabriela A L; Lopes, Viviane C P; Mainardi, Pedro H; Dos Santos, Juliana A; de Azevedo Duarte, Lidia; Otero, Igor V R; da Silva Yoshida, Aline M; Feitosa, Valker A; Pessoa, Adalberto; Sette, Lara D

    2015-01-01

    The ocean is considered to be a great reservoir of biodiversity. Microbial communities in marine environments are ecologically relevant as intermediaries of energy, and play an important role in nutrient regeneration cycles as decomposers of dead and decaying organic matter. In this sense, marine-derived fungi can be considered as a source of enzymes of industrial and/or environmental interest. Fungal strains isolated from different substrates, such as invertebrates, decaying wood, seawater, sediments, and mangrove detritus, have been reported to be producers of hydrolytic and/or oxidative enzymes, with alginate lyase, amylase, cellulase, chitinase, glucosidase, inulinase, keratinase, ligninase, lipase, nuclease, phytase, protease, and xylanase being among the enzymes produced by fungi of marine origin. These enzymes present temperature and pH optima ranging from 35 to 70(∘)C, and 3.0 to 11.0, respectively. High-level production in bioreactors is mainly performed using submerged-state fermentation. Certain marine-derived fungal strains present enzymes with alkaline and cold-activity characteristics, and salinity is considered an important condition in screening and production processes. The adaptability of marine-derived fungi to oceanic conditions can be considered an attractive point in the field of fungal marine biotechnology. In this review, we focus on the advances in discovering enzymes from marine-derived fungi and their biotechnological relevance.

  4. Marine-derived fungi: diversity of enzymes and biotechnological applications

    Science.gov (United States)

    Bonugli-Santos, Rafaella C.; dos Santos Vasconcelos, Maria R.; Passarini, Michel R. Z.; Vieira, Gabriela A. L.; Lopes, Viviane C. P.; Mainardi, Pedro H.; dos Santos, Juliana A.; de Azevedo Duarte, Lidia; Otero, Igor V. R.; da Silva Yoshida, Aline M.; Feitosa, Valker A.; Pessoa, Adalberto; Sette, Lara D.

    2015-01-01

    The ocean is considered to be a great reservoir of biodiversity. Microbial communities in marine environments are ecologically relevant as intermediaries of energy, and play an important role in nutrient regeneration cycles as decomposers of dead and decaying organic matter. In this sense, marine-derived fungi can be considered as a source of enzymes of industrial and/or environmental interest. Fungal strains isolated from different substrates, such as invertebrates, decaying wood, seawater, sediments, and mangrove detritus, have been reported to be producers of hydrolytic and/or oxidative enzymes, with alginate lyase, amylase, cellulase, chitinase, glucosidase, inulinase, keratinase, ligninase, lipase, nuclease, phytase, protease, and xylanase being among the enzymes produced by fungi of marine origin. These enzymes present temperature and pH optima ranging from 35 to 70∘C, and 3.0 to 11.0, respectively. High-level production in bioreactors is mainly performed using submerged-state fermentation. Certain marine-derived fungal strains present enzymes with alkaline and cold-activity characteristics, and salinity is considered an important condition in screening and production processes. The adaptability of marine-derived fungi to oceanic conditions can be considered an attractive point in the field of fungal marine biotechnology. In this review, we focus on the advances in discovering enzymes from marine-derived fungi and their biotechnological relevance. PMID:25914680

  5. Marine-derived fungi: diversity of enzymes and biotechnological applications

    Directory of Open Access Journals (Sweden)

    Rafaella Costa Bonugli-Santos

    2015-04-01

    Full Text Available The ocean is considered to be a great reservoir of biodiversity. Microbial communities in marine environments are ecologically relevant as intermediaries of energy, and play an important role in nutrient regeneration cycles as decomposers of dead and decaying organic matter. In this sense, marine-derived fungi can be considered as a source of enzymes of industrial and/or environmental interest. Fungal strains isolated from different substrates, such as invertebrates, decaying wood, seawater, sediments and mangrove detritus, have been reported to be producers of hydrolytic and/or oxidative enzymes, with alginate lyase, amylase, cellulase, chitinase, glucosidase, inulinase, keratinase, ligninase, lipase, nuclease, phytase, protease and xylanase being among the enzymes produced by fungi of marine origin. These enzymes present temperature and pH optima ranging from 35 to 70 ºC, and 3.0 to 11.0, respectively. High-level production in bioreactors is mainly performed using submerged-state fermentation. Certain marine-derived fungal strains present enzymes with alkaline and cold-activity characteristics, and salinity is considered an important condition in screening and production processes. The adaptability of marine-derived fungi to oceanic conditions can be considered an attractive point in the field of fungal marine biotechnology. In this review, we focus on the advances in discovering enzymes from marine-derived fungi and their biotechnological relevance.

  6. [Fungal biomass estimation in soils from southwestern Buenos Aires province (Argentina) using calcofluor white stain].

    Science.gov (United States)

    Vázquez, María B; Amodeo, Martín R; Bianchinotti, María V

    Soil microorganisms are vital for ecosystem functioning because of the role they play in soil nutrient cycling. Agricultural practices and the intensification of land use have a negative effect on microbial activities and fungal biomass has been widely used as an indicator of soil health. The aim of this study was to analyze fungal biomass in soils from southwestern Buenos Aires province using direct fluorescent staining and to contribute to its use as an indicator of environmental changes in the ecosystem as well as to define its sensitivity to weather conditions. Soil samples were collected during two consecutive years. Soil smears were prepared and stained with two different concentrations of calcofluor, and the fungal biomass was estimated under an epifluorescence microscope. Soil fungal biomass varied between 2.23 and 26.89μg fungal C/g soil, being these values in the range expected for the studied soil type. The fungal biomass was positively related to temperature and precipitations. The methodology used was reliable, standardized and sensitive to weather conditions. The results of this study contribute information to evaluate fungal biomass in different soil types and support its use as an indicator of soil health for analyzing the impact of different agricultural practices. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  7. Fungal keratitis: The Aravind experience

    Science.gov (United States)

    Prajna, Venkatesh N; Prajna, Lalitha; Muthiah, Srinivasan

    2017-01-01

    Research becomes very significant and meaningful when it addresses a significant public health problem of a region. Fungal keratitis is a serious problem affecting the agrarian poor and hence requires attention from public health specialists. The approach to a public health issue should focus not only on treatment but also prevention or at least show a significant thrust to reduce the morbidity of the problem. At our institution, we have developed a special interest in fungal keratitis and tried to study it in a multitude of aspects. As we put the pieces of the puzzle together, we believe that interest will be rekindled among policymakers, clinicians, microbiologists, pharmaceutical industry, and basic scientists to work together to join forces and take up an integrative approach to managing this problem. It is also believed that the article underscores the need and importance of having a focused approach to ensuring a successful career in clinical research. PMID:29044053

  8. A plant pathology perspective of fungal genome sequencing.

    Science.gov (United States)

    Aylward, Janneke; Steenkamp, Emma T; Dreyer, Léanne L; Roets, Francois; Wingfield, Brenda D; Wingfield, Michael J

    2017-06-01

    The majority of plant pathogens are fungi and many of these adversely affect food security. This mini-review aims to provide an analysis of the plant pathogenic fungi for which genome sequences are publically available, to assess their general genome characteristics, and to consider how genomics has impacted plant pathology. A list of sequenced fungal species was assembled, the taxonomy of all species verified, and the potential reason for sequencing each of the species considered. The genomes of 1090 fungal species are currently (October 2016) in the public domain and this number is rapidly rising. Pathogenic species comprised the largest category (35.5 %) and, amongst these, plant pathogens are predominant. Of the 191 plant pathogenic fungal species with available genomes, 61.3 % cause diseases on food crops, more than half of which are staple crops. The genomes of plant pathogens are slightly larger than those of other fungal species sequenced to date and they contain fewer coding sequences in relation to their genome size. Both of these factors can be attributed to the expansion of repeat elements. Sequenced genomes of plant pathogens provide blueprints from which potential virulence factors were identified and from which genes associated with different pathogenic strategies could be predicted. Genome sequences have also made it possible to evaluate adaptability of pathogen genomes and genomic regions that experience selection pressures. Some genomic patterns, however, remain poorly understood and plant pathogen genomes alone are not sufficient to unravel complex pathogen-host interactions. Genomes, therefore, cannot replace experimental studies that can be complex and tedious. Ultimately, the most promising application lies in using fungal plant pathogen genomics to inform disease management and risk assessment strategies. This will ultimately minimize the risks of future disease outbreaks and assist in preparation for emerging pathogen outbreaks.

  9. Recommendations for Risk Categorization and Prophylaxis of Invasive Fungal Diseases in Hematological Malignancies: A Critical Review of Evidence and Expert Opinion (TEO-4)

    OpenAIRE

    Can Boğa; Zahit Bolaman; Seçkin Çağırgan; İhsan Karadoğan; Mehmet Ali Özcan; Fahir Özkalemkaş; Rabin Saba; Mehmet Sönmez; Esin Şenol; Hamdi Akan; Murat Akova

    2015-01-01

    This is the last of a series of articles on invasive fungal infections prepared by opinion leaders in Turkey. The aim of these articles is to guide clinicians in managing invasive fungal diseases in hematological malignancies and stem cell transplantation based on the available best evidence in this field. The previous articles summarized the diagnosis and treatment of invasive fungal disease and this article aims to explain the risk categorization and guide the antifungal prophylaxis in inva...

  10. Fungal genome resources at NCBI

    Science.gov (United States)

    Robbertse, B.; Tatusova, T.

    2011-01-01

    The National Center for Biotechnology Information (NCBI) is well known for the nucleotide sequence archive, GenBank and sequence analysis tool BLAST. However, NCBI integrates many types of biomolecular data from variety of sources and makes it available to the scientific community as interactive web resources as well as organized releases of bulk data. These tools are available to explore and compare fungal genomes. Searching all databases with Fungi [organism] at http://www.ncbi.nlm.nih.gov/ is the quickest way to find resources of interest with fungal entries. Some tools though are resources specific and can be indirectly accessed from a particular database in the Entrez system. These include graphical viewers and comparative analysis tools such as TaxPlot, TaxMap and UniGene DDD (found via UniGene Homepage). Gene and BioProject pages also serve as portals to external data such as community annotation websites, BioGrid and UniProt. There are many different ways of accessing genomic data at NCBI. Depending on the focus and goal of research projects or the level of interest, a user would select a particular route for accessing genomic databases and resources. This review article describes methods of accessing fungal genome data and provides examples that illustrate the use of analysis tools. PMID:22737589

  11. Systems biology of fungal infection

    Directory of Open Access Journals (Sweden)

    Fabian eHorn

    2012-04-01

    Full Text Available Elucidation of pathogenicity mechanisms of the most important human pathogenic fungi, Aspergillus fumigatus and Candida albicans, has gained great interest in the light of the steadily increasing number of cases of invasive fungal infections.A key feature of these infections is the interaction of the different fungal morphotypes with epithelial and immune effector cells in the human host. Because of the high level of complexity, it is necessary to describe and understand invasive fungal infection by taking a systems biological approach, i.e., by a comprehensive quantitative analysis of the non-linear and selective interactions of a large number of functionally diverse, and frequently multifunctional, sets of elements, e.g., genes, proteins, metabolites, which produce coherent and emergent behaviours in time and space. The recent advances in systems biology will now make it possible to uncover the structure and dynamics of molecular and cellular cause-effect relationships within these pathogenic interactions.We review current efforts to integrate omics and image-based data of host-pathogen interactions into network and spatio-temporal models. The modelling will help to elucidate pathogenicity mechanisms and to identify diagnostic biomarkers and potential drug targets for therapy and could thus pave the way for novel intervention strategies based on novel antifungal drugs and cell therapy.

  12. Evaluation of fungal laccase immobilized on natural nanostructured bacterial cellulose

    Directory of Open Access Journals (Sweden)

    Lin eChen

    2015-11-01

    Full Text Available The aim of this work was to assess the possibility of using native bacterial nanocellulose (BC as a carrier for laccase immobilization. BC was synthesized by Gluconacetobacter xylinus, which was statically cultivated in a mannitol-based medium and was freeze-dried to form BC sponge after purification. For the first time, fungal laccase from Trametes versicolor was immobilized on the native nanofibril network-structured BC sponge through physical adsorption and cross-linking with glutaraldehyde. The properties including morphologic and structural features of the BC as well as the immobilized enzyme were thoroughly investigated. It was found that enzyme immobilized by cross-linking exhibited broader pH operation range of high catalytic activity as well as higher running stability compared to free and adsorbed enzyme. Using ABTS as substrate, the optimum pH value was 3.5 for the adsorption-immobilized laccase and 4.0 for the crosslinking-immobilized laccase. The immobilized enzyme retained 69% of the original activity after being recycled 7 times. Novel applications of the BC-immobilized enzyme tentatively include active packaging, construction of biosensors, and establishment of bioreactors.

  13. Epidemiological characteristics and laboratory diagnosis of fungal keratitis. A three-year study

    Directory of Open Access Journals (Sweden)

    Bharathi Jayahar

    2003-01-01

    Full Text Available Purpose: To study the epidemiological characteristics and laboratory diagnosis of fungal keratitis seen at a tertiary eye care referral centre in South India. Methods: A retrospective review of all culture-proven fungal keratitis seen over a 3-year period, September 1999 through August 2002. Results: Fungal aetiology were confirmed in1095(34.4% of 3183 corneal ulcers. The predominant fungal species isolated was Fusarium spp (471; 42.82% followed by Aspergillus spp (286; 26%. Males (712; 65.08% were more often affected (P< 0.0001. A large proportion of the patients (732; 66.85% were in the younger age group (21 to 50 years. A majority (879; 80.27% came from rural areas (P Conclusion: Agricultural activity and related ocular trauma were principal causes of mycotic keratitis. A potassium hydroxide (KOH wet mount preparation is a simple, and sensitive, method for diagnosis.

  14. Links between plant and fungal diversity in habitat fragments of coastal shrubland.

    Directory of Open Access Journals (Sweden)

    Mia R Maltz

    Full Text Available Habitat fragmentation is widespread across ecosystems, detrimentally affecting biodiversity. Although most habitat fragmentation studies have been conducted on macroscopic organisms, microbial communities and fungal processes may also be threatened by fragmentation. This study investigated whether fragmentation, and the effects of fragmentation on plants, altered fungal diversity and function within a fragmented shrubland in southern California. Using fluorimetric techniques, we assayed enzymes from plant litter collected from fragments of varying sizes to investigate enzymatic responses to fragmentation. To isolate the effects of plant richness from those of fragment size on fungi, we deployed litter bags containing different levels of plant litter diversity into the largest fragment and incubated in the field for one year. Following field incubation, we determined litter mass loss and conducted molecular analyses of fungal communities. We found that leaf-litter enzyme activity declined in smaller habitat fragments with less diverse vegetation. Moreover, we detected greater litter mass loss in litter bags containing more diverse plant litter. Additionally, bags with greater plant litter diversity harbored greater numbers of fungal taxa. These findings suggest that both plant litter resources and fungal function may be affected by habitat fragmentation's constraints on plants, possibly because plant species differ chemically, and may thus decompose at different rates. Diverse plant assemblages may produce a greater variety of litter resources and provide more ecological niche space, which may support greater numbers of fungal taxa. Thus, reduced plant diversity may constrain both fungal taxa richness and decomposition in fragmented coastal shrublands. Altogether, our findings provide evidence that even fungi may be affected by human-driven habitat fragmentation via direct effects of fragmentation on plants. Our findings underscore the importance

  15. Links between plant and fungal diversity in habitat fragments of coastal shrubland.

    Science.gov (United States)

    Maltz, Mia R; Treseder, Kathleen K; McGuire, Krista L

    2017-01-01

    Habitat fragmentation is widespread across ecosystems, detrimentally affecting biodiversity. Although most habitat fragmentation studies have been conducted on macroscopic organisms, microbial communities and fungal processes may also be threatened by fragmentation. This study investigated whether fragmentation, and the effects of fragmentation on plants, altered fungal diversity and function within a fragmented shrubland in southern California. Using fluorimetric techniques, we assayed enzymes from plant litter collected from fragments of varying sizes to investigate enzymatic responses to fragmentation. To isolate the effects of plant richness from those of fragment size on fungi, we deployed litter bags containing different levels of plant litter diversity into the largest fragment and incubated in the field for one year. Following field incubation, we determined litter mass loss and conducted molecular analyses of fungal communities. We found that leaf-litter enzyme activity declined in smaller habitat fragments with less diverse vegetation. Moreover, we detected greater litter mass loss in litter bags containing more diverse plant litter. Additionally, bags with greater plant litter diversity harbored greater numbers of fungal taxa. These findings suggest that both plant litter resources and fungal function may be affected by habitat fragmentation's constraints on plants, possibly because plant species differ chemically, and may thus decompose at different rates. Diverse plant assemblages may produce a greater variety of litter resources and provide more ecological niche space, which may support greater numbers of fungal taxa. Thus, reduced plant diversity may constrain both fungal taxa richness and decomposition in fragmented coastal shrublands. Altogether, our findings provide evidence that even fungi may be affected by human-driven habitat fragmentation via direct effects of fragmentation on plants. Our findings underscore the importance of restoring

  16. Expansion of the aspartate [beta]-semialdehyde dehydrogenase family: the first structure of a fungal ortholog

    Energy Technology Data Exchange (ETDEWEB)

    Arachea, B.T.; Liu, X.; Pavlovsky, A.G.; Viola, R.E. (Toledo)

    2010-08-13

    The enzyme aspartate semialdehyde dehydrogenase (ASADH) catalyzes a critical transformation that produces the first branch-point intermediate in an essential microbial amino-acid biosynthetic pathway. The first structure of an ASADH isolated from a fungal species (Candida albicans) has been determined as a complex with its pyridine nucleotide cofactor. This enzyme is a functional dimer, with a similar overall fold and domain organization to the structurally characterized bacterial ASADHs. However, there are differences in the secondary-structural elements and in cofactor binding that are likely to cause the lower catalytic efficiency of this fungal enzyme. Alterations in the dimer interface, through deletion of a helical subdomain and replacement of amino acids that participate in a hydrogen-bonding network, interrupt the intersubunit-communication channels required to support an alternating-site catalytic mechanism. The detailed functional information derived from this new structure will allow an assessment of ASADH as a possible target for antifungal drug development.

  17. Actinobacterial chitinase-like enzymes: profiles of rhizosphere versus non-rhizosphere isolates.

    Science.gov (United States)

    Gonzalez-Franco, Ana C; Deobald, Lee A; Spivak, Aaron; Crawford, Don L

    2003-11-01

    The objective of this study was to determine if antifungal actinomycetes isolated from rhizosphere and non-rhizosphere soils exhibit different chitinase-like production and (or) induction patterns. Selected isolates from both habitats were compared. Chitinase-like levels and isoform characteristic patterns were evaluated over time in culture fluids of isolates grown on media containing different combinations of colloidal chitin and fungal cell wall (FCW) preparation. Supernatants were also subjected to native and non-native polyacrylamide gel electrophoresis (PAGE), using glycol chitin amended gels. For non-native PAGE, protein samples were denatured by two different approaches. Multiple active bands, ranging from 20 to 53 kDa and present in varying amounts, were detected in gels for most strains. Different substrate preferences were observed among strains, and different chitinase-like enzymes were produced, depending upon the substrate combinations used. The presence of FCW in the medium induced specific chitinase-like enzymes not observed otherwise. Enzymatic activities and profiles of the isolates, however, were strain and substrate specific rather than habitat specific. However, a sagebrush rhizosphere soil had a larger actinomycete community with higher chitinolytic activities than the nearby bulk soil. The use of PAGE to compare chitinase-like proteins induced in media with and without FCW was useful for identifying chitinase-like enzymes potentially involved in antifungal activity.

  18. Serious fungal infections in Egypt.

    Science.gov (United States)

    Zaki, S M; Denning, D W

    2017-06-01

    We aimed to estimate the burden of serious fungal infections in Egypt, currently unknown, based on the size of the populations at risk and available epidemiological data. Data were obtained from the World Health Organization (WHO), the Joint United Nations Programme on HIV/AIDS (UNAIDS), and published reports with clearcut denominators. When no data existed, risk populations were used to estimate frequencies of fungal infections, using previously described methodology. The population of Egypt in 2011 was ∼82,500,000; 31% children, and 8% women >60 years of age. Amongst about 21.8 million women aged 15-50 years, recurrent vulvovaginal candidiasis (≥4 episodes/year) is estimated to occur in 1.3 million (3,169/100,000 females). Using a low international average rate of 5/100,000, we estimate 4,127 cases of candidaemia, and 619 patients with intra-abdominal candidiasis. Amongst the survivors of pulmonary tuberculosis (TB) in Egypt in 2012, 319 new cases of chronic pulmonary aspergillosis (CPA) are likely, a prevalence of 1,005 post-TB and a total prevalence estimate of 3,015 CPA patients in all. Asthma is common in Egypt, affecting 9.4% of adults, 5.35 million, and so ABPA and SAFS were estimated in around 162/100,000 and 214/100,000 respectively. Invasive aspergillosis is estimated to affect 495 patients following leukaemia therapy, there are an estimated 37 cases in renal and liver transplant recipients, and an estimated 132 patients develop IA in the context of lung cancer. Amongst 641,000 COPD admissions to hospital each year, 8,337 patients develop IA. The total HIV-infected population is small, with an estimated 6,500 patients, 2,500 not on antiretroviral therapy. Amongst HIV-infected patients, 38 (0.6%) cases of cryptococcal meningitis and 125 (1.9%) cases of Pneumocystis pneumonia are estimated each year. Fungal keratitis is common, with 28-55% (mean 40%) of corneal infections being fungal, an estimated total of 11,550 cases. The present study indicates

  19. Phylogenetic distribution of fungal sterols.

    Directory of Open Access Journals (Sweden)

    John D Weete

    Full Text Available BACKGROUND: Ergosterol has been considered the "fungal sterol" for almost 125 years; however, additional sterol data superimposed on a recent molecular phylogeny of kingdom Fungi reveals a different and more complex situation. METHODOLOGY/PRINCIPAL FINDINGS: The interpretation of sterol distribution data in a modern phylogenetic context indicates that there is a clear trend from cholesterol and other Delta(5 sterols in the earliest diverging fungal species to ergosterol in later diverging fungi. There are, however, deviations from this pattern in certain clades. Sterols of the diverse zoosporic and zygosporic forms exhibit structural diversity with cholesterol and 24-ethyl -Delta(5 sterols in zoosporic taxa, and 24-methyl sterols in zygosporic fungi. For example, each of the three monophyletic lineages of zygosporic fungi has distinctive major sterols, ergosterol in Mucorales, 22-dihydroergosterol in Dimargaritales, Harpellales, and Kickxellales (DHK clade, and 24-methyl cholesterol in Entomophthorales. Other departures from ergosterol as the dominant sterol include: 24-ethyl cholesterol in Glomeromycota, 24-ethyl cholest-7-enol and 24-ethyl-cholesta-7,24(28-dienol in rust fungi, brassicasterol in Taphrinales and hypogeous pezizalean species, and cholesterol in Pneumocystis. CONCLUSIONS/SIGNIFICANCE: Five dominant end products of sterol biosynthesis (cholesterol, ergosterol, 24-methyl cholesterol, 24-ethyl cholesterol, brassicasterol, and intermediates in the formation of 24-ethyl cholesterol, are major sterols in 175 species of Fungi. Although most fungi in the most speciose clades have ergosterol as a major sterol, sterols are more varied than currently understood, and their distribution supports certain clades of Fungi in current fungal phylogenies. In addition to the intellectual importance of understanding evolution of sterol synthesis in fungi, there is practical importance because certain antifungal drugs (e.g., azoles target reactions in

  20. Enzyme-MOF (metal-organic framework) composites.

    Science.gov (United States)

    Lian, Xizhen; Fang, Yu; Joseph, Elizabeth; Wang, Qi; Li, Jialuo; Banerjee, Sayan; Lollar, Christina; Wang, Xuan; Zhou, Hong-Cai

    2017-06-06

    The ex vivo application of enzymes in various processes, especially via enzyme immobilization techniques, has been extensively studied in recent years in order to enhance the recyclability of enzymes, to minimize enzyme contamination in the product, and to explore novel horizons for enzymes in biomedical applications. Possessing remarkable amenability in structural design of the frameworks as well as almost unparalelled surface tunability, Metal-Organic Frameworks (MOFs) have been gaining popularity as candidates for enzyme immobilization platforms. Many MOF-enzyme composites have achieved unprecedented results, far outperforming free enzymes in many aspects. This review summarizes recent developments of MOF-enzyme composites with special emphasis on preparative techniques and the synergistic effects of enzymes and MOFs. The applications of MOF-enzyme composites, primarily in transferation, catalysis and sensing, are presented as well. The enhancement of enzymatic activity of the composites over free enzymes in biologically incompatible conditions is emphasized in many cases.

  1. Preparation of human drug metabolites using fungal peroxygenases

    Science.gov (United States)

    Marzena Poraj-Kobielska; Matthias Kinne; René Ullrich; Katrin Scheibner; Gernot Kayser; Kenneth E. Hammel; Martin Hofrichter

    2011-01-01

    The synthesis of hydroxylated and O- or N-dealkylated human drug metabolites (HDMs) via selective monooxygenation remains a challenging task for synthetic organic chemists. Here we report that aromatic peroxygenases (APOs; EC 1.11.2.1) secreted by the agaric fungi Agrocybe aegerita and Coprinellus...

  2. Industrially Important Carbohydrate Degrading Enzymes from Yeasts: Pectinases, Chitinases, and β-1,3-Glucanases

    Science.gov (United States)

    Gummadi, Sathyanarayana N.; Kumar, D. Sunil; Dash, Swati S.; Sahu, Santosh Kumar

    Polysaccharide degrading enzymes are hydrolytic enzymes, which have a lot of industrial potential and also play a crucial role in carbon recycling. Pectinases, chitinases and glucanases are the three major polysaccharide degrading enzymes found abundantly in nature and these enzymes are mainly produced by fungal strains. Production of these enzymes by yeasts is advantageous over fungi, because the former are easily amenable to genetic manipulations and time required for growth and production is less than that of the latter. Several yeasts belonging to Saccharomyces, Pichia, Rhodotorula and Cryptococcus produce extracellular pectinases, glucanases and chitinases. This chapter emphasizes on the biological significance of these enzymes, their production and their industrial applications.

  3. Targeted Gene Replacement in Fungal Pathogens via Agrobacterium tumefaciens- Mediated Transformation

    DEFF Research Database (Denmark)

    Frandsen, Rasmus John Normand; Frandsen, Mette; Giese, Nanna Henriette

    2012-01-01

    cloning without the need for subcloning. The cloning efficiency is not always as high as desired, but it still presents an efficient alternative to restriction enzyme and ligase-based cloning systems. The USER technology offers a higher four fragment cloning efficiency than In-Fusion, but depends...... on specific structures in the binary vector. The available fungal binary vectors adapted for the USER system are described and protocols are provided for vector design and construction. A general protocol for verification of the resulting gene replacement events in the recipient fungal cells is also given...

  4. Invasive fungal infections after natural disasters.

    Science.gov (United States)

    Benedict, Kaitlin; Park, Benjamin J

    2014-03-01

    The link between natural disasters and subsequent fungal infections in disaster-affected persons has been increasingly recognized. Fungal respiratory conditions associated with disasters include coccidioidomycosis, and fungi are among several organisms that can cause near-drowning pneumonia. Wound contamination with organic matter can lead to post-disaster skin and soft tissue fungal infections, notably mucormycosis. The role of climate change in the environmental growth, distribution, and dispersal mechanisms of pathogenic fungi is not fully understood; however, ongoing climate change could lead to increased disaster-associated fungal infections. Fungal infections are an often-overlooked clinical and public health issue, and increased awareness by health care providers, public health professionals, and community members regarding disaster-associated fungal infections is needed.

  5. Innovative production of fungal pulp from Trametes versicolor and its application in a fungal paper box containing clove oil.

    Science.gov (United States)

    Srikaew, Benyapa; Matan, Narumol; Aewsiri, Tanong

    2017-09-01

    The aims of this study were to develop fungal pulp from Trametes versicolor (white-rot fungi) and apply it with clove oil in a paper box to inhibit mold growth on the surface of peanuts. Broken rice media with different sugar solutions (2-10% w w -1 ) were prepared and then inoculated with T. versicolor mycelium at amounts from 0.5 to 1.5% w w -1 . Fungal pulp and commercial paper (50 g) at different ratios (100:0, 70:30, 50:50, 30:70 and 0:100) were mixed and prepared before being placed into a stainless box (5 cm long by 5 cm wide). For the antimicrobial activity against Aspergillus flavus on peanuts, a paper box was incorporated with 2.5, 5 and 7.5% w w -1 of clove oil, eugenol, caryophyllene, and a combination of eugenol and caryophyllene at ratios of 7:1, 4:4, and 1:7. Results indicated that the highest fungal pulp biomass of T. versicolor in broken rice media was found when using 6% sugar with 1% mycelium inoculums. Fungal pulp and commercial paper at the ratio of 70:30 produced the highest value of hardness. The paper box containing clove oil at 7.5% w w -1 inhibited A. flavus on peanuts for at least 28 days while the control had mold growth within 3 days. Combining eugenol and caryophyllene, the main components of clove oil, at the ratio of 7:1 (7.5% w w -1 ) in the paper box should be a key factor to inhibit A. flavus during storage.

  6. Active invasion of bacteria into living fungal cells

    Science.gov (United States)

    Moebius, Nadine; Üzüm, Zerrin; Dijksterhuis, Jan; Lackner, Gerald; Hertweck, Christian

    2014-01-01

    The rice seedling blight fungus Rhizopus microsporus and its endosymbiont Burkholderia rhizoxinica form an unusual, highly specific alliance to produce the highly potent antimitotic phytotoxin rhizoxin. Yet, it has remained a riddle how bacteria invade the fungal cells. Genome mining for potential symbiosis factors and functional analyses revealed that a type 2 secretion system (T2SS) of the bacterial endosymbiont is required for the formation of the endosymbiosis. Comparative proteome analyses show that the T2SS releases chitinolytic enzymes (chitinase, chitosanase) and chitin-binding proteins. The genes responsible for chitinolytic proteins and T2SS components are highly expressed during infection. Through targeted gene knock-outs, sporulation assays and microscopic investigations we found that chitinase is essential for bacteria to enter hyphae. Unprecedented snapshots of the traceless bacterial intrusion were obtained using cryo-electron microscopy. Beyond unveiling the pivotal role of chitinolytic enzymes in the active invasion of a fungus by bacteria, these findings grant unprecedented insight into the fungal cell wall penetration and symbiosis formation. DOI: http://dx.doi.org/10.7554/eLife.03007.001 PMID:25182414

  7. Widespread occurrence of expressed fungal secretory peroxidases in forest soils.

    Science.gov (United States)

    Kellner, Harald; Luis, Patricia; Pecyna, Marek J; Barbi, Florian; Kapturska, Danuta; Krüger, Dirk; Zak, Donald R; Marmeisse, Roland; Vandenbol, Micheline; Hofrichter, Martin

    2014-01-01

    Fungal secretory peroxidases mediate fundamental ecological functions in the conversion and degradation of plant biomass. Many of these enzymes have strong oxidizing activities towards aromatic compounds and are involved in the degradation of plant cell wall (lignin) and humus. They comprise three major groups: class II peroxidases (including lignin peroxidase, manganese peroxidase, versatile peroxidase and generic peroxidase), dye-decolorizing peroxidases, and heme-thiolate peroxidases (e.g. unspecific/aromatic peroxygenase, chloroperoxidase). Here, we have repeatedly observed a widespread expression of all major peroxidase groups in leaf and needle litter across a range of forest ecosystems (e.g. Fagus, Picea, Acer, Quercus, and Populus spp.), which are widespread in Europe and North America. Manganese peroxidases and unspecific peroxygenases were found expressed in all nine investigated forest sites, and dye-decolorizing peroxidases were observed in five of the nine sites, thereby indicating biological significance of these enzymes for fungal physiology and ecosystem processes. Transcripts of selected secretory peroxidase genes were also analyzed in pure cultures of several litter-decomposing species and other fungi. Using this information, we were able to match, in environmental litter samples, two manganese peroxidase sequences to Mycena galopus and Mycena epipterygia and one unspecific peroxygenase transcript to Mycena galopus, suggesting an important role of this litter- and coarse woody debris-dwelling genus in the disintegration and transformation of litter aromatics and organic matter formation.

  8. Widespread occurrence of expressed fungal secretory peroxidases in forest soils.

    Directory of Open Access Journals (Sweden)

    Harald Kellner

    Full Text Available Fungal secretory peroxidases mediate fundamental ecological functions in the conversion and degradation of plant biomass. Many of these enzymes have strong oxidizing activities towards aromatic compounds and are involved in the degradation of plant cell wall (lignin and humus. They comprise three major groups: class II peroxidases (including lignin peroxidase, manganese peroxidase, versatile peroxidase and generic peroxidase, dye-decolorizing peroxidases, and heme-thiolate peroxidases (e.g. unspecific/aromatic peroxygenase, chloroperoxidase. Here, we have repeatedly observed a widespread expression of all major peroxidase groups in leaf and needle litter across a range of forest ecosystems (e.g. Fagus, Picea, Acer, Quercus, and Populus spp., which are widespread in Europe and North America. Manganese peroxidases and unspecific peroxygenases were found expressed in all nine investigated forest sites, and dye-decolorizing peroxidases were observed in five of the nine sites, thereby indicating biological significance of these enzymes for fungal physiology and ecosystem processes. Transcripts of selected secretory peroxidase genes were also analyzed in pure cultures of several litter-decomposing species and other fungi. Using this information, we were able to match, in environmental litter samples, two manganese peroxidase sequences to Mycena galopus and Mycena epipterygia and one unspecific peroxygenase transcript to Mycena galopus, suggesting an important role of this litter- and coarse woody debris-dwelling genus in the disintegration and transformation of litter aromatics and organic matter formation.

  9. Fungal catalases: function, phylogenetic origin and structure.

    Science.gov (United States)

    Hansberg, Wilhelm; Salas-Lizana, Rodolfo; Domínguez, Laura

    2012-09-15

    Most fungi have several monofunctional heme-catalases. Filamentous ascomycetes (Pezizomycotina) have two types of large-size subunit catalases (L1 and L2). L2-type are usually induced by different stressors and are extracellular enzymes; those from the L1-type are not inducible and accumulate in asexual spores. L2 catalases are important for growth and the start of cell differentiation, while L1 are required for spore germination. In addition, pezizomycetes have one to four small-size subunit catalases. Yeasts (Saccharomycotina) do not have large-subunit catalases and generally have one peroxisomal and one cytosolic small-subunit catalase. Small-subunit catalases are inhibited by substrate while large-subunit catalases are activated by H(2)O(2). Some small-subunit catalases bind NADPH preventing inhibition by substrate. We present a phylogenetic analysis revealing one or two events of horizontal gene transfers from Actinobacteria to a fungal ancestor before fungal diversification, as the origin of large-size subunit catalases. Other possible horizontal transfers of small- and large-subunit catalases genes were detected and one from bacteria to the fungus Malassezia globosa was analyzed in detail. All L2-type catalases analyzed presented a secretion signal peptide. Mucorales preserved only L2-type catalases, with one containing a secretion signal if two or more are present. Basidiomycetes have only L1-type catalases, all lacking signal peptide. Fungal small-size catalases are related to animal catalases and probably evolved from a common ancestor. However, there are several groups of small-size catalases. In particular, a conserved group of fungal sequences resemble plant catalases, whose phylogenetic origin was traced to a group of bacteria. This group probably has the heme orientation of plant catalases and could in principle bind NADPH. From almost a hundred small-subunit catalases only one fourth has a peroxisomal localization signal and in fact many fungi lack

  10. Fungal Endocarditis: Update on Diagnosis and Management.

    Science.gov (United States)

    Pasha, Ahmed Khurshid; Lee, Justin Z; Low, See-Wei; Desai, Hem; Lee, Kwan S; Al Mohajer, Mayar

    2016-10-01

    Fungal endocarditis is an extremely debilitating disease associated with high morbidity and mortality. Candida spp. are the most common isolated organisms in fungal endocarditis. It is most prevalent in patients who are immunosuppressed and intravenous drug users. Most patients present with constitutional symptoms, which are indistinguishable from bacterial endocarditis, hence a high index of suspicion is required for pursuing diagnosis. Diagnosis of fungal endocarditis can be very challenging: most of the time, blood cultures are negative or take a long time to yield growth. Fungal endocarditis mandates an aggressive treatment strategy. A medical and surgical combined approach is the cornerstone of therapy. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Enzyme activity assay of glycoprotein enzymes based on a boronate affinity molecularly imprinted 96-well microplate.

    Science.gov (United States)

    Bi, Xiaodong; Liu, Zhen

    2014-12-16

    Enzyme activity assay is an important method in clinical diagnostics. However, conventional enzyme activity assay suffers from apparent interference from the sample matrix. Herein, we present a new format of enzyme activity assay that can effectively eliminate the effects of the sample matrix. The key is a 96-well microplate modified with molecularly imprinted polymer (MIP) prepared according to a newly proposed method called boronate affinity-based oriented surface imprinting. Alkaline phosphatase (ALP), a glycoprotein enzyme that has been routinely used as an indicator for several diseases in clinical tests, was taken as a representative target enzyme. The prepared MIP exhibited strong affinity toward the template enzyme (with a dissociation constant of 10(-10) M) as well as superb tolerance for interference. Thus, the enzyme molecules in a complicated sample matrix could be specifically captured and cleaned up for enzyme activity assay, which eliminated the interference from the sample matrix. On the other hand, because the boronate affinity MIP could well retain the enzymatic activity of glycoprotein enzymes, the enzyme captured by the MIP was directly used for activity assay. Thus, additional assay time and possible enzyme or activity loss due to an enzyme release step required by other methods were avoided. Assay of ALP in human serum was successfully demonstrated, suggesting a promising prospect of the proposed method in real-world applications.

  12. Enzyme detection by microfluidics

    DEFF Research Database (Denmark)

    2013-01-01

    Microfluidic-implemented methods of detecting an enzyme, in particular a DNA-modifying enzyme, are provided, as well as methods for detecting a cell, or a microorganism expressing said enzyme. The enzyme is detected by providing a nucleic acid substrate, which is specifically targeted...... by that enzyme...

  13. Bacterial and fungal chitinase chiJ orthologs evolve under different selective constraints following horizontal gene transfer

    Directory of Open Access Journals (Sweden)

    Ubhayasekera Wimal

    2012-10-01

    Full Text Available Abstract Background Certain bacteria from the genus Streptomyces are currently used as biological control agents against plant pathogenic fungi. Hydrolytic enzymes that degrade fungal cell wall components, such as chitinases, are suggested as one possible mechanism in biocontrol interactions. Adaptive evolution of chitinases are previously reported for plant chitinases involved in defence against fungal pathogens, and in fungal chitinases involved in fungal-fungal interactions. In this study we investigated the molecular evolution of chitinase chiJ in the bacterial genus Streptomyces. In addition, as chiJ orthologs are previously reported in certain fungal species as a result from horizontal gene transfer, we conducted a comparative study of differences in evolutionary patterns between bacterial and fungal taxa. Findings ChiJ contained three sites evolving under strong positive selection and four groups of co-evolving sites. Regions of high amino acid diversity were predicted to be surface-exposed and associated with coil regions that connect certain α-helices and β-strands in the family 18 chitinase TIM barrel structure, but not associated with the catalytic cleft. The comparative study with fungal ChiJ orthologs identified three regions that display signs of type 1 functional divergence, where unique adaptations in the bacterial and fungal taxa are driven by positive selection. Conclusions The identified surface-exposed regions of chitinase ChiJ where sequence diversification is driven by positive selection may putatively be related to functional divergence between bacterial and fungal orthologs. These results show that ChiJ orthologs have evolved under different selective constraints following the horizontal gene transfer event.

  14. Review article: fungal microbiota and digestive diseases.

    Science.gov (United States)

    Wang, Z K; Yang, Y S; Stefka, A T; Sun, G; Peng, L H

    2014-04-01

    The role of the fungal microbiota in digestive diseases is poorly defined, but is becoming better understood due to advances in metagenomics. To review the gastrointestinal fungal microbiota and its relationship with digestive diseases. Search of the literature using PubMed and MEDLINE databases. Subject headings including 'fungal-bacterial interactions', 'mycotoxins', 'immunity to fungi', 'fungal infection', 'fungal microbiota', 'mycobiome' and 'digestive diseases' were used. The fungal microbiota is an integral part of the gastrointestinal microecosystem with up to 10(6) microorganisms per gram of faeces. Next-generation sequencing of the fungal 18S rRNA gene has allowed better characterisation of the gastrointestinal mycobiome. Numerous interactions between fungi and bacteria and the complex immune response to gastrointestinal commensal or pathogenic fungi all impact on the pathophysiology of inflammatory bowel disease and other gastrointestinal inflammatory entities such as peptic ulcers. Mycotoxins generated as fungal metabolites contribute to disturbances of gastrointestinal barrier and immune functions and are associated with chronic intestinal inflammatory conditions as well as hepatocellular and oesophagogastric cancer. Systemic and gastrointestinal disease can also lead to secondary fungal infections. Fungal genomic databases and methodologies need to be further developed and will allow a much better understanding of the diversity and function of the mycobiome in gastrointestinal inflammation, tumourigenesis, liver cirrhosis and transplantation, and its alteration as a consequence of antibiotic therapy and chemotherapy. The fungal microbiota and its metabolites impact gastrointestinal function and contribute to the pathogenesis of digestive diseases. Further metagenomic analyses of the gastrointestinal mycobiome in health and disease is needed. © 2014 John Wiley & Sons Ltd.

  15. Elevated Liver Enzymes

    Science.gov (United States)

    Symptoms Elevated liver enzymes By Mayo Clinic Staff Elevated liver enzymes may indicate inflammation or damage to cells in the liver. Inflamed or ... than normal amounts of certain chemicals, including liver enzymes, into the bloodstream, which can result in elevated ...

  16. Genomic insights into the fungal lignocellulolytic system of Myceliophthora thermophila

    Directory of Open Access Journals (Sweden)

    Anthi eKarnaouri

    2014-06-01

    Full Text Available The microbial conversion of solid cellulosic biomass to liquid biofuels may provide a renewable energy source for transportation fuels. Cellulolytic fungi represent a promising group of organisms, as they have evolved complex systems for adaptation to their natural habitat. The filamentous fungus Myceliophthora thermophila constitutes an exceptionally powerful cellulolytic microorganism that synthesizes a complete set of enzymes necessary for the breakdown of plant cell wall. The genome of this fungus has been recently sequenced and annotated, allowing systematic examination and identification of enzymes required for the degradation of lignocellulosic biomass. The genomic analysis revealed the existence of an expanded enzymatic repertoire including numerous cellulases, hemicellulases and enzymes with auxiliary activities, covering the most of the recognized CAZy families. Most of them were predicted to possess a secretion signal and undergo through post translational glycosylation modifications. These data offer a better understanding of activities embedded in fungal lignocellulose decomposition mechanisms and suggest that M. thermophila could be made usable as an industrial production host for cellulolytic and hemicellulolytic enzymes.

  17. The importance of sourcing enzymes from non-conventional fungi for metabolic engineering and biomass breakdown.

    Science.gov (United States)

    Seppälä, Susanna; Wilken, St Elmo; Knop, Doriv; Solomon, Kevin V; O'Malley, Michelle A

    2017-11-01

    A wealth of fungal enzymes has been identified from nature, which continue to drive strain engineering and bioprocessing for a range of industries. However, while a number of clades have been investigated, the vast majority of the fungal kingdom remains unexplored for industrial applications. Here, we discuss selected classes of fungal enzymes that are currently in biotechnological use, and explore more basal, non-conventional fungi and their underexploited biomass-degrading mechanisms as promising agents in the transition towards a bio-based society. Of special interest are anaerobic fungi like the Neocallimastigomycota, which were recently found to harbor the largest diversity of biomass-degrading enzymes among the fungal kingdom. Enzymes sourced from these basal fungi have been used to metabolically engineer substrate utilization in yeast, and may offer new paths to lignin breakdown and tunneled biocatalysis. We also contrast classic enzymology approaches with emerging 'omics'-based tools to decipher function within novel fungal isolates and identify new promising enzymes. Recent developments in genome editing are expected to accelerate discovery and metabolic engineering within these systems, yet are still limited by a lack of high-resolution genomes, gene regulatory regions, and even appropriate culture conditions. Finally, we present new opportunities to harness the biomass-degrading potential of undercharacterized fungi via heterologous expression and engineered microbial consortia. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  18. Fungal infections of the orbit

    Directory of Open Access Journals (Sweden)

    Bipasha Mukherjee

    2016-01-01

    Full Text Available Fungal infections of the orbit can lead to grave complications. Although the primary site of inoculation of the infective organism is frequently the sinuses, the patients can initially present to the ophthalmologist with ocular signs and symptoms. Due to its varied and nonspecific clinical features, especially in the early stages, patients are frequently misdiagnosed and even treated with steroids which worsen the situation leading to dire consequences. Ophthalmologists should be familiar with the clinical spectrum of disease and the variable presentation of this infection, as early diagnosis and rapid institution of appropriate therapy are crucial elements in the management of this invasive sino-orbital infection. In this review, relevant clinical, microbiological, and imaging findings are discussed along with the current consensus on local and systemic management. We review the recent literature and provide a comprehensive analysis. In the immunocompromised, as well as in healthy patients, a high index of suspicion must be maintained as delay in diagnosis of fungal pathology may lead to disfiguring morbidity or even mortality. Obtaining adequate diagnostic material for pathological and microbiological examination is critical. Newer methods of therapy, particularly oral voriconazole and topical amphotericin B, may be beneficial in selected patients.

  19. Fungal transmission of plant viruses.

    Science.gov (United States)

    Campbell, R N

    1996-01-01

    Thirty soilborne viruses or virus-like agents are transmitted by five species of fungal vectors. Ten polyhedral viruses, of which nine are in the family Tombusviridae, are acquired in the in vitro manner and do not occur within the resting spores of their vectors, Olpidium brassicae and O. bornovanus. Fungal vectors for other viruses in the family should be sought even though tombusviruses are reputed to be soil transmitted without a vector. Eighteen rod-shaped viruses belonging to the furo- and bymovirus groups and to an unclassified group are acquired in the in vivo manner and survive within the resting spores of their vector, O. brassicae, Polymyxa graminis, P. betae, and Spongospora subterranea. The viral coat protein has an essential role in in vitro transmission. With in vivo transmission a site in the coat protein-read through protein (CP-RT) of beet necrotic yellow vein furovirus determines vector transmissibility as does a site in a similar 98-kDa polyprotein of barley mild mosaic bymovirus. The mechanisms by which virions move (or are moved) into and out of the protoplasm of zoospores or of thalli needs study.

  20. Immune response to fungal infection.

    Science.gov (United States)

    Diamond, R D

    1989-01-01

    In general, fungi are saprophytes that are well adapted to grow in nature supported by diverse nutritional substrates. For fungi, in contrast to many other microorganisms that infect humans, parasitism is an accidental phenomenon rather than an obligatory requirement for survival. Thus, with progressive improvement in our capabilities to prolong survival of patients with global defects in host defense mechanisms, clinical experience suggests that human tissues may support growth of numerous species of saprophytic fungi that share the capacity to grow at 37 degrees C. Normally, however, a broad array of natural and acquired host defense mechanisms make the occurrence of progressive, systemic, life-threatening mycoses extremely rare events. When one or another of these host defense mechanisms is compromised, one of a variety of significant fungal infections may then progress. Mycoses may be broadly categorized into those controlled largely by natural cellular defenses vs. acquired cell-mediated immunity. Notwithstanding data that permit such general classification of host factors controlling one or another invasive mycosis, the diverse structural and antigenic properties of individual fungi create unique patterns of infections in individual, characteristic host settings. Thus, while some broad generalizations are possible, definition of predisposing factors for specific individual mycoses (and, ultimately, prospects for corrective immunotherapy) requires careful characterization of diverse features of fungal forms mediating divergent immune responses.

  1. Optimal Fungal Space Searching Algorithms.

    Science.gov (United States)

    Asenova, Elitsa; Lin, Hsin-Yu; Fu, Eileen; Nicolau, Dan V; Nicolau, Dan V

    2016-10-01

    Previous experiments have shown that fungi use an efficient natural algorithm for searching the space available for their growth in micro-confined networks, e.g., mazes. This natural "master" algorithm, which comprises two "slave" sub-algorithms, i.e., collision-induced branching and directional memory, has been shown to be more efficient than alternatives, with one, or the other, or both sub-algorithms turned off. In contrast, the present contribution compares the performance of the fungal natural algorithm against several standard artificial homologues. It was found that the space-searching fungal algorithm consistently outperforms uninformed algorithms, such as Depth-First-Search (DFS). Furthermore, while the natural algorithm is inferior to informed ones, such as A*, this under-performance does not importantly increase with the increase of the size of the maze. These findings suggest that a systematic effort of harvesting the natural space searching algorithms used by microorganisms is warranted and possibly overdue. These natural algorithms, if efficient, can be reverse-engineered for graph and tree search strategies.

  2. Solamargine production by a fungal endophyte of Solanum nigrum.

    Science.gov (United States)

    El-Hawary, S S; Mohammed, R; AbouZid, S F; Bakeer, W; Ebel, R; Sayed, A M; Rateb, M E

    2016-04-01

    The aim was to isolate, identify and characterize endophytes from Solanum nigrum L. as a new source of the cytotoxic steroidal alkaloid solamargine. Three endophytic fungi; SNFSt, SNFL and SNFF were isolated from S. nigrum and identified by molecular methods. Preliminary TLC screening showed a common metabolite between the plant and one of these fungi, SNFSt which was identified as Aspergillus flavus based on the phylogenetic analysis of its ITS sequence. Subsequent LC-HRESIMS analysis unambiguously established the identity of the compound based on its molecular formula and its characteristic MS(2) fragmentation pattern as solamargine. To ascertain its identity, fungal solamargine was isolated using preparative TLC and its structure was fully characterized using NMR spectroscopic techniques and high-resolution mass spectrometric analysis. Solamargine production could be followed and quantified for a total of 11 generations of this fungus with a titer of ~250-300 μg l(-1) . This study represents one of the first examples where host plant-derived compounds have been demonstrated to be steadily produced by an endophytic fungi in sizeable quantities. The production of solamargine (found in the host plant) by a cultivable fungal endophyte at a significant yield is a new observation. Further experiments such as media optimization, OSMAC (One Strain Many Compounds) or epigenetic modifiers could be applied to enhance the fungal solamargine production. The endophytic fungus SNFSt isolated from S. nigrum may be utilized for quantitative production of the potent cytotoxic metabolite solamargine. © 2016 The Society for Applied Microbiology.

  3. Fungal infection knowledge gap in Ethiopia

    African Journals Online (AJOL)

    EPHA USER33

    receiving immunosuppressive therapy, and patients with chronic obstructive lung disease (1). Fungi also play a role in allergic fungal disease such as allergic broncho- pulmonary Aspergilosis (ABPA) and chronic or deep tissue infections. The laboratory diagnosis of fungal infection starts with a simple potassium hydroxide.

  4. Tropospheric ozone as a fungal elicitor

    Indian Academy of Sciences (India)

    Tropospheric ozone has been proven to trigger biochemical plant responses that are similar to the ones induced by an attack of fungal pathogens, i.e. it resembles fungal elicitors. This suggests that ozone can represent a valid tool for the study of stress responses and induction of resistance to pathogens. This review ...

  5. Common fungal diseases of Russian forests

    Science.gov (United States)

    Evgeny P. Kuz' michevl; Ella s. Sokolova; Elena G. Kulikova

    2001-01-01

    Describes common fungal diseases of Russian forests, including diagnostic signs and symptoms, pathogen biology, damage caused by the disease, and methods of control. The fungal diseases are divided into two groups: those that are the most common in Russian forests and those that are found only in Russia. Within each group, diseases are subdivided by plant organ...

  6. DEGRADATION OF ZEARALENONE BY LACCASE ENZYME

    Directory of Open Access Journals (Sweden)

    IULIANA BANU

    2014-01-01

    Full Text Available The degradation of zearalenone by pure fungal laccase (Trametes versicolor was investigated in this study. In the experiments different activities of laccase and different initial zearalenone concentrations from liquid medium were used. At 0.4 mg∙mL-1 laccase concentration, after 240 min time of incubation, was obtained an 81.7 % reduction of zearalenone from liquid medium. The zearalenone degradation depends on the initial concentration of mycotoxin and on the activity of laccase. The results obtained can contribute to the development of preventative strategies to reduce mycotoxin contamination of food by involving enzymes.

  7. ANTI-FUNGAL ACTIVITIES OF m-IODOBENZOIC ACID AND SOME ...

    African Journals Online (AJOL)

    Although the use of these metals complexes as food preservatives may only be fungi-static and not fungi-toxic, their use in bread preparation might extend the shelf life of bread from 24 hours to 96 hours. KEY WORDS: Anti-fungal activities, Alkali metal iodobenzoates, Alkaline earth metal iodobenzoates, m-Iodobenzoic acid ...

  8. Global changes alter soil fungal communities and alter rates of organic matter decomposition

    Science.gov (United States)

    Moore, J.; Frey, S. D.

    2016-12-01

    Global changes - such as warming, more frequent and severe droughts, increasing atmospheric CO2, and increasing nitrogen (N) deposition rates - are altering ecosystem processes. The balance between soil carbon (C) accumulation and decomposition is determined in large part by the activity and biomass of detrital organisms, namely soil fungi, and yet their sensitivity to global changes remains unresolved. We present results from a meta-analysis of 200+ studies spanning manipulative and observational field experiments to quantify fungal responses to global change and expected consequences for ecosystem C dynamics. Warming altered the functional soil microbial community by reducing the ratio of fungi to bacteria (f:b) total fungal biomass. Additionally, warming reduced lignolytic enzyme activity generally by one-third. Simulated N deposition affected f:b differently than warming, but the effect on fungal biomass and activity was similar. The effect of N-enrichment on f:b was contingent upon ecosystem type; f:b increased in alpine meadows and heathlands but decreased in temperate forests following N-enrichment. Across ecosystems, fungal biomass marginally declined by 8% in N-enriched soils. In general, N-enrichment reduced fungal lignolytic enzyme activity, which could explain why soil C accumulates in some ecosystems following warming and N-enrichment. Several global change experiments have reported the surprising result that soil C builds up following increases in temperature and N deposition rates. While site-specific studies have examined the role of soil fungi in ecosystem responses to global change, we present the first meta-analysis documenting general patterns of global change impacts on soil fungal communities, biomass, and activity. In sum, we provide evidence that soil microbial community shifts and activity plays a large part in ecosystem responses to global changes, and have the potential to alter the magnitude of the C-climate feedback.

  9. 21 CFR 184.1027 - Mixed carbohydrase and protease enzyme product.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Mixed carbohydrase and protease enzyme product. 184... enzyme product. (a) Mixed carbohydrase and protease enzyme product is an enzyme preparation that includes... current good manufacturing practice conditions of use: (1) The ingredient is used as an enzyme, as defined...

  10. INCIDENCE OF FUNGAL ELEMENTS IN SINONASAL POLYPOSIS

    Directory of Open Access Journals (Sweden)

    Santhosh G. S

    2016-12-01

    Full Text Available BACKGROUND Nasal polyposis is a disease entity characterised by formation of pseudoedema of sinonasal mucus membrane progressing to form polyps. It presents clinically with nasal obstruction and fleshy masses in the nasal cavity. The nasal mucosa reacts to formation of polypi in allergic fungal sinusitis also. The present study is an attempt to demonstrate possible fungal elements from the polypi removed during surgery by KOH study and HPE study. The aim of the study is to find out the incidence of fungal elements in sinonasal polyposis. MATERIALS AND METHODS 50 patients attending the ENT OPD for nasal obstruction and showing polypi on anterior rhinoscopy were selected. All the patients were subjected to surgery and specimens collected were subjected to KOH study and histopathology to demonstrate fungal elements. RESULTS Among 50 patients, the age range was from 9-57 years; mean age- 36.46 years. The male-to-female ratio was 1.5:1. Deviated nasal septum was found in 38% of patients. Among the unilateral cases, 47% were antrochoanal polyps and 53% were ethmoid polyps. Out of 50 patients, only 3 specimens were positive for fungal elements with KOH study and only 2 cases with fungal culture. Thus, the incidence of fungal elements in sinonasal polyposis was 6%. CONCLUSION The incidence of fungal elements in sinonasal polyposis was 6%. Histopathological examination of polypectomy specimen was negative for invasive fungal disease and showed inflammatory changes only. There is no difference in the detection of the presence of fungal by two methods.

  11. Chemoenzymatic preparation of germacrene analogues.

    Science.gov (United States)

    Cascón, Oscar; Touchet, Sabrina; Miller, David J; Gonzalez, Veronica; Faraldos, Juan A; Allemann, Rudolf K

    2012-10-09

    A small library of novel germacrenes was generated using a combination of two plant enzymes, germacrene A synthase, and D synthase and modified farnesyl diphosphate (FDP) analogues. This chemoenzymatic approach allows the preparation of potentially valuable volatiles for biological studies.

  12. Chemoenzymatic preparation of germacrene analogues

    OpenAIRE

    Cascón, Oscar; Touchet, Sabrina; Miller, David James; Gonzalez, Veronica; Faraldos, Juan A.; Allemann, Rudolf Konrad

    2012-01-01

    A small library of novel germacrenes was generated using a combination of two plant enzymes, germacrene A synthase, and D synthase and modified farnesyl diphosphate (FDP) analogues. This chemoenzymatic approach allows the preparation of potentially valuable volatiles for biological studies.

  13. Depolymerization of chitosan by enzymes from the digestive tract of ...

    African Journals Online (AJOL)

    A complex of enzymes was isolated in a preparation derived from the digestive tract of sea cucumber, Stichopus japonicus. Hydrolysis of chitosan using this enzyme preparation decreased its molecular weight (Mw), increased its water solubility and produced water-soluble chitosan (WSC). The conditions for hydrolysis were ...

  14. Some factors including radiation affecting the productivity of proteinase enzymes by mucor lamprosporus

    International Nuclear Information System (INIS)

    El-Kabbany, H.M.I.

    1996-01-01

    In the present time, great attention has been focused on the production of milk clotting enzymes from microbial source for use as remain substitute due to the increasing demands on rennin for cheese making and the prohibition of the slaughter of small calves. The present investigation included the isolation and identification of remin-like enzyme fungal producers from different egyptian food and soil samples. Different factors including gamma radiation affecting the capability of selected isolate to produce the enzyme was also included. Special attention has also given to study the effect of different purification methods of the produced enzyme. The properties of the purified enzyme were also investigated

  15. Quantification of Fungal DNA by Using Fluorescence Resonance Energy Transfer and the Light Cycler System

    Science.gov (United States)

    Loeffler, Juergen; Henke, Norbert; Hebart, Holger; Schmidt, Diethard; Hagmeyer, Lars; Schumacher, Ulrike; Einsele, Hermann

    2000-01-01

    The Light Cycler technique combines rapid in vitro amplification of DNA in glass capillaries with real-time species determination and quantification of DNA load. We have established a quantitative PCR protocol for two clinically important pathogens, Candida albicans and Aspergillus fumigatus. The sensitivity of the assay was comparable to those of previously described PCR protocols (5 CFU/ml). Specific detection of C. albicans and A. fumigatus could be achieved. The assay showed a high reproducibility of 96 to 99%. The assay was linear in a range between 101 and 104 Aspergillus conidia. As capillaries do not have to be reopened for post-PCR analysis, the risk of carryover contaminations could be minimized. The Light Cycler allowed quantification of the fungal loads in a limited number of clinical specimens from patients with hematological malignancies and histologically proven invasive fungal infections. Five of nine positive samples had fungal loads between 5 and 10 CFU/ml of blood, two of nine positive samples had fungal loads between 10 and 100 CFU/ml of blood, and two of nine samples had fungal loads of more than 100 CFU/ml of blood. All samples were also found to be PCR positive by PCR–enzyme-linked immunosorbent assay analysis. PMID:10655350

  16. Chapter 8: Invasive fungal rhinosinusitis.

    Science.gov (United States)

    Duggal, Praveen; Wise, Sarah K

    2013-01-01

    Invasive fungal rhinosinusitis (IFRS) is a disease of the paranasal sinuses and nasal cavity that typically affects immunocompromised patients in the acute fulminant form. Early symptoms can often mimic rhinosinusitis, while late symptoms can cause significant morbidity and mortality. Swelling and mucosal thickening can quickly progress to pale or necrotic tissue in the nasal cavity and sinuses, and the disease can rapidly spread and invade the palate, orbit, cavernous sinus, cranial nerves, skull base, carotid artery, and brain. IFRS can be life threatening if left undiagnosed or untreated. While the acute fulminant form of IFRS is the most rapidly progressive and destructive, granulomatous and chronic forms also exist. Diagnosis of IFRS often mandates imaging studies in conjunction with clinical, endoscopic, and histopathological examination. Treatment of IFRS consists of reversing the underlying immunosuppression, antifungal therapy, and aggressive surgical debridement. With early diagnosis and treatment, IFRS can be treated and increase patient survival.

  17. Chapter 8: Invasive fungal rhinosinusitis.

    Science.gov (United States)

    Duggal, Praveen; Wise, Sarah K

    2013-05-01

    Invasive fungal rhinosinusitis (IFRS) is a disease of the paranasal sinuses and nasal cavity that typically affects immunocompromised patients in the acute fulminant form. Early symptoms can often mimic rhinosinusitis, while late symptoms can cause significant morbidity and mortality. Swelling and mucosal thickening can quickly progress to pale or necrotic tissue in the nasal cavity and sinuses, and the disease can rapidly spread and invade the palate, orbit, cavernous sinus, cranial nerves, skull base, carotid artery, and brain. IFRS can be life threatening if left undiagnosed or untreated. While the acute fulminant form of IFRS is the most rapidly progressive and destructive, granulomatous and chronic forms also exist. Diagnosis of IFRS often mandates imaging studies in conjunction with clinical, endoscopic, and histopathological examination. Treatment of IFRS consists of reversing the underlying immunosuppression, antifungal therapy, and aggressive surgical debridement. With early diagnosis and treatment, IFRS can be treated and increase patient survival.

  18. Conversion of ammonia-pretreated switchgrass to biofuel precursors by bacterial-fungal consortia under solid-state and submerged-state cultivation.

    Science.gov (United States)

    Jain, A; Pelle, H S; Baughman, W H; Henson, J M

    2017-04-01

    The aim of this study was to develop and evaluate bacterial-fungal communities to deconstruct switchgrass to biofuel precursors. Bacterial-fungal consortia, mesophilic (25°C) and thermophilic (50°C), were enriched from switchgrass bales from which enzyme mixtures were used to deconstruct delignified switchgrass (DSG). The bacterial-fungal consortia were able to produce enzymes including endoglucanase, exoglucanase, β-glucosidase, xylanase, xylosidase and pectinase to convert DSG to soluble carbohydrates. 454 pyrosequencing revealed that Paenibacillus and Streptomyces were the dominant bacteria in the mesophilic and thermophilic consortia respectively. Penicillium and Acremonium were the dominant fungi in the mesophilic consortia, whereas Aspergillus and Penicillium were the dominant fungi present in the thermophilic consortia. The results show that the state of cultivation, solid-state or submerged-state, affects the community structure as well as enzyme activities produced by these bacterial-fungal consortia. The enzyme mixture produced by the bacterial-fungal consortia released a higher amount of xylose than glucose during saccharification of DSG. The study provides a novel approach to produce enzymes for conversion of lignocellulolytic feedstocks to soluble sugars which can be used to produce biofuel precursors. © 2016 The Society for Applied Microbiology.

  19. Biocatalytic Single Enzyme Nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Grate, Jay W.; Kim, Jungbae

    2004-03-31

    As an innovative way of enzyme stabilization, we recently developed a new enzyme composite of nano-meter scale that we call "single-enzyme nanoparticles (SENs)" (9). Each enzyme molecule is surrounded with a porous composite organic/inorganic network of less than a few nanometers think. This approach represents a new type of enzyme-containing nanostructure. In experiments with perotease (chymotrypsin, CT), the activity of single enzyme nanoparticle form of the enzyme was greatly stabilized compared to the free form, without imposing a serious mass transfer limitation of substrates. In this chapter we will describe the synthesis, characterization and catalytic activity of the new SENs.

  20. Fungal infection in organ transplant patients.

    Science.gov (United States)

    Hong, Wei; Wen, Hai; Liao, Wanqing

    2003-09-01

    To review the characteristics and evolution of the fungal spectrum, and the risk factors causing fungal infection, and to make progress in diagnosing fungal infection after organ transplantation. An English-language literature search (MEDLINE 1990 - 2000) and bibliographic review of textbooks and review articles. Twenty-three articles were selected from the literature that specifically addressed the stated purpose. Fungal infections in organ transplant patients were generally divided into two types: (1) disseminated primary or reactivation infection with one of the geographically restricted systemic mycoses; (2) opportunistic infection by fungal species that rarely cause invasive infection in normal hosts. The risk factors of fungal infection after a transplant can be evaluated and predicted according to the organ recipient's conditions before, during and after the transplant. Progress in early diagnostic methods during the past 10 years has mainly revolved around two aspects, culture and non-culture. It is important to undertake a systemic evaluation on the condition of the organ recipient before, during and after a transplant; should any risk factor for fungal infection be suspected, diagnosis should be made as early as possible by employing mycological techniques including culture and non-culture methods.

  1. Bioethanol production from brewers spent grains using a fungal consolidated bioprocessing (CBP) approach

    OpenAIRE

    Wilkinson, Stuart; Smart, Katherine A.; James, Sue; Cook, David J.

    2017-01-01

    Production of bioethanol from brewers spent grains (BSG) using consolidated bioprocessing (CBP) is reported. Each CBP system consists of a primary filamentous fungal species, which secretes the enzymes required to deconstruct biomass, paired with a secondary yeast species to ferment liberated sugars to ethanol. Interestingly, although several pairings of fungi were investigated, the sake fermentation system (A.oryzae and S.cerevisiae NCYC479) was found to yield the highest concentrations of e...

  2. Synthesis, Anti-Bacterial and Anti-Fungal Evaluation of Pyrazoline Derivatives

    Directory of Open Access Journals (Sweden)

    Ashvin D. Panchal

    2012-01-01

    Full Text Available The Series of N-(5-(Substituted phenyl-4,5-dihydro-1H-pyrazol-3-yl-4H-1,2,4-triazol-4-amine compounds were prepared by reaction of 4-amino-1,2,4-Triazole with Acetyl Chloride followed by different aromatic aldehydes and cyclization with hydrazine hydrate. The structures of new compounds were confirmed by IR and 1H-NMR spectral data. Anti-bacterial and Anti-fungal activities were evaluated and compared with the standard drugs, some compounds of the series exhibited promising anti-microbial and anti-fungal activity compared to standard drugs.

  3. Fungal colonization of air-conditioning systems

    Directory of Open Access Journals (Sweden)

    Ljaljević-Grbić Milica

    2008-01-01

    Full Text Available Fungi have been implicated as quantitatively the most important bioaerosol component of indoor air associated with contaminated air-conditioning systems. rarely, indoor fungi may cause human infections, but more commonly allergenic responses ranging from pneumonitis to asthma-like symptoms. From all air conditioner filters analyzed, 16 fungal taxa were isolated and identified. Aspergillus fumigatus causes more lethal infections worldwide than any other mold. Air-conditioning filters that adsorb moisture and volatile organics appear to provide suitable substrates for fungal colonization. It is important to stress that fungal colonization of air-conditioning systems should not be ignored, especially in hospital environments.

  4. Fungal Infections From Human and Animal Contact

    Directory of Open Access Journals (Sweden)

    Dennis J. Baumgardner

    2017-04-01

    Full Text Available Fungal infections in humans resulting from human or animal contact are relatively uncommon, but they include a significant proportion of dermatophyte infections. Some of the most commonly encountered diseases of the integument are dermatomycoses. Human or animal contact may be the source of all types of tinea infections, occasional candidal infections, and some other types of superficial or deep fungal infections. This narrative review focuses on the epidemiology, clinical features, diagnosis and treatment of anthropophilic dermatophyte infections primarily found in North America. Other human-acquired and zoonotic fungal infections also are discussed in brief.

  5. Enzyme Production and Nitrogen Fixation by Free, Immobilized and Coimmobilized Inoculants of Trichoderma harzianum and Azospirillum brasilense and Their Possible Role in Growth Promotion of Tomato

    Directory of Open Access Journals (Sweden)

    Momein H. El-Katatny

    2010-01-01

    Full Text Available A plant growth-promoting rhizobacterium (Azospirillum brasilense strain Az and a biocontrol fungus (Trichoderma harzianum strain T24 have been evaluated for their individual and combined production of hydrolytic enzymes, nitrogen fixation and their possible role in growth promotion of tomato seedlings. The studied organisms were inoculated as free or calcium alginate-encapsulated cells. All freshly prepared macrobeads showed high encapsulation capacity (EC/% of inocula compared with dry macrobeads. Results of enzyme production did not exhibit consistent pattern of the effect of encapsulation process on enzyme production. Beads entrapping bacterial and/or fungal cells were used successfully in 3 repeated cycles in the presence of fresh sterile culture medium in each growth cycle. Enzyme production by immobilized bacterial and/or fungal cells increased as the growth cycles were repeated. Co-culturing of A. brasilense with T. harzianum (free or immobilized in semisolid nitrogen deficient medium (N-free medium enabled A. brasilense to fix nitrogen on pectin, chitin and carboxymethyl cellulose. The activity of nitrogen fixation by A. brasilense in the case of single and combined cultures with Trichoderma (using dry encapsulated beads into the sterile soil increased with the addition of carbon source. Most of inoculations with free or alginate macrobead formulations of T. harzianum and/or A. brasilense showed significant increase in the growth parameters of tomato seedlings. The root system grew more profusely in the case of all seeds treated with A. brasilense. The growth parameters of Az/T24-treated seeds using dry coimmobilized macrobeads were higher than those of the untreated control. Moreover, the effect was improved significantly in soil enriched with different C sources. Enhanced tomato seedling growth after the co-inoculation could be due to the synergistic effect of both Trichoderma and Azospirillum. Finally, co-inoculation with Azospirillum

  6. A comprehensive overview on genomically directed assembly of aromatic polyketides and macrolide lactones using fungal megasynthases.

    Science.gov (United States)

    Saruwatari, Takayoshi; Praseuth, Alex P; Sato, Michio; Torikai, Kohei; Noguchi, Hiroshi; Watanabe, Kenji

    2011-01-01

    Fungal polyketide synthases (PKSs) catalyze a carbon-carbon bond forming reaction in an iterative manner using a variety of acyl-CoA molecules as substrates when biosynthesizing complex polyketides. Although most members from this class of natural products exhibit notable biological activities, often they are naturally produced in trace levels or cultivation of the analyte-producing organism is less than feasible. Appropriately, to contend with the former challenge, one must identify any translational bottleneck and perform functional analysis of the associated enzymes. In recent years, many gene clusters purportedly responsible for biosynthesizing polyketides have been identified and cataloged from a variety of fungal genomes including genes coding for iterative PKSs, particulary bikaverin, zearalenone and hypothemycin biosynthetic enzymes. Mounting appreciation of these highly specific codons and their translational consequence will afford scientists the ability to anticipate the fungal metabolite by correlating an organism's genomic cluster to an appropriate biosynthetic system. It was observed in recent reports, the successful production of these recombinant enzymes using an Escherichia coli expression system which in turn conferred the anticipated metabolite in vitro. This review will focus on iterative PKSs responsible for biosynthesizing bikaverin, zearalenone and hypothemycin, and expand on befitting enzymatic reaction mechanisms and development of a highly versatile system that could potentially generate biologically active compounds.

  7. Novel fungal proteins in the chalkbrood infection of honey bee larvae

    DEFF Research Database (Denmark)

    Roth, Doris; Jensen, Annette Bruun; Grell, Morten Nedergaard

    2009-01-01

    . Here we investigate the interaction between the honey bee and its fungal pathogen Ascosphaera apis, the causative agent of chalkbrood, by identifying enzymes secreted by bee and fungus during different timepoints of infection. Upon testing A. apis-infected larvae for enzyme activity, the larvae...... the trappants are sequenced and annotated, selected genes are further described. As a result, we will deepen the understanding of chalkbrood, one of the main honey bee pests with relevant impact on the economy, among others due to the essential role of bees in pollination....

  8. Expanding Fungal Diets Through Synthetic Algal-Fungal Mutualism

    Science.gov (United States)

    Sharma, Alaisha; Galazka, Jonathan (Editor)

    2015-01-01

    Fungi can synthesize numerous molecules with important properties, and could be valuable production platforms for space exploration and colonization. However, as heterotrophs, fungi require reduced carbon. This limits their efficiency in locations such as Mars, where reduced carbon is scarce. We propose a system to induce mutualistic symbiosis between the green algae Chlamydomonas reinhardtii and the filamentous fungi Neurospora crassa. This arrangement would mimic natural algal-fungal relationships found in lichens, but have added advantages including increased growth rate and genetic tractability. N. crassa would metabolize citrate (C6H5O7 (sup -3)) and release carbon dioxide (CO2) that C. reinhardtii would assimilate into organic sugars during photosynthesis. C. reinhardtii would metabolize nitrate (NO3-) and release ammonia (NH3) as a nitrogen source for N. crassa. A N. crassa mutant incapable of reducing nitrate will be used to force this interaction. This system eliminates the need to directly supply its participants with carbon dioxide and ammonia. Furthermore, the release of oxygen by C. reinhardtii via photosynthesis would enable N. crassa to respire. We hope to eventually create a system closer to lichen, in which the algae transfers not only nitrogen but reduced carbon, as organic sugars, to the fungus for growth and production of valuable compounds.

  9. Oxidative Stress Responses in the Human Fungal Pathogen, Candida albicans

    Science.gov (United States)

    da Silva Dantas, Alessandra; Day, Alison; Ikeh, Mélanie; Kos, Iaroslava; Achan, Beatrice; Quinn, Janet

    2015-01-01

    Candida albicans is a major fungal pathogen of humans, causing approximately 400,000 life-threatening systemic infections world-wide each year in severely immunocompromised patients. An important fungicidal mechanism employed by innate immune cells involves the generation of toxic reactive oxygen species (ROS), such as superoxide and hydrogen peroxide. Consequently, there is much interest in the strategies employed by C. albicans to evade the oxidative killing by macrophages and neutrophils. Our understanding of how C. albicans senses and responds to ROS has significantly increased in recent years. Key findings include the observations that hydrogen peroxide triggers the filamentation of this polymorphic fungus and that a superoxide dismutase enzyme with a novel mode of action is expressed at the cell surface of C. albicans. Furthermore, recent studies have indicated that combinations of the chemical stresses generated by phagocytes can actively prevent C. albicans oxidative stress responses through a mechanism termed the stress pathway interference. In this review, we present an up-date of our current understanding of the role and regulation of oxidative stress responses in this important human fungal pathogen. PMID:25723552

  10. Synergistic effect of Aspergillus niger and Trichoderma reesei enzyme sets on the saccharification of wheat straw and sugarcane bagasse

    NARCIS (Netherlands)

    van den Brink, J.; Maitan-Alfenas, Gabriela Piccolo; Zou, Gen; Wang, Chengshu; Zhou, Zhihua; Guimarães, Valéria Monteze; de Vries, Ronald P

    2014-01-01

    Plant-degrading enzymes can be produced by fungi on abundantly available low-cost plant biomass. However, enzymes sets after growth on complex substrates need to be better understood, especially with emphasis on differences between fungal species and the influence of inhibitory compounds in plant

  11. Trametes suaveolens as ligninolytic enzyme producer

    Directory of Open Access Journals (Sweden)

    Knežević Aleksandar

    2013-01-01

    Full Text Available Species of the genus Trametes represent one of the most efficient lignin-degraders which can be attributed to a well developed ligninolytic enzyme system. Current trends are screening of ability of new species to produce these enzymes, as well as the optimization of conditions for their overproduction. Therefore, the aim of the study was to evaluate the potential of T. suaveolens to synthesize laccase and Mn-oxidizing peroxidases during fermentation of the selected plant raw materials. Level of enzyme activities was measured on 7, 10 and 14th day of submersion, as well as the solid-state fermentation of wheat straw and oak sawdust in the presence of NH4NO3 in previously determined optimal nitrogen concentration of 25 mM. The enzyme activity was determined spectrophotometrically using ABTS and phenol red as the substrates. The highest level of laccase activity (1087.1 U/L was noted after 7 days of wheat straw solid-state fermentation, while during the submerged cultivation the production of the enzyme was not noted. Submerged cultivation in oak sawdust-enriched medium was the optimal for activity of Mn-dependent peroxidase (1767.7 U/L on day 14 and Mn-independent peroxidase (1113.7 U/L on day 7. Introduction of T. suaveolens to produce ligninolytic enzyme represented the base for further study, as well as the determination of relation between enzyme activity and rate of lignin degradation. It could lead to greater possibility of fungal species selection with high delignification capacity, which could take participation in sustainable production of food, feed, fibres, and energy, environmentally friendly pollution prevention, and bioremediation.

  12. PNNL Fungal Biotechnology Core DOE-OBP Project

    Energy Technology Data Exchange (ETDEWEB)

    Baker, Scott E.; Bruno, Kenneth S.; Butcher, Mark G.; Collett, James R.; Culley, David E.; Dai, Ziyu; Magnuson, Jon K.; Panisko, Ellen A.

    2009-11-30

    In 2009, we continued to address barriers to fungal fermentation in the primary areas of morphology control, genomics, proteomics, fungal hyperproductivity, biomass-to-products via fungal based consolidated bioprocesses, and filamentous fungal ethanol. “Alternative renewable fuels from fungi” was added as a new subtask. Plans were also made to launch a new advanced strain development subtask in FY2010.

  13. Saccharification of sunflower stalks using lignocellulases from a fungal consortium comprising Pholiota adiposa and Armillaria gemina.

    Science.gov (United States)

    Ramachandran, Priyadharshini; Kim, Tae-Su; Dhiman, Saurabh Sudha; Li, Jinglin; Park, Ji-Hyun; Choi, Joon-Ho; Kim, Jae Young; Kim, Dongwook; Lee, Jung-Kul

    2015-09-01

    Lignocellulases from Armillaria gemina and Pholiota adiposa are efficient in hydrolyzing aspen and poplar biomass, respectively. In the present study, lignocellulosic enzymes obtained from a fungal consortium comprising P. adiposa and A. gemina were used for the saccharification of sunflower stalks. Sunflower stalks were thermochemically pretreated using 2 % NaOH at 50 °C for 24 h. The saccharification process parameters including substrate concentration, enzyme loading, pH, and temperature were optimized using response surface methodology to improve the saccharification yield. The highest enzymatic hydrolysis (84.3 %) was obtained using the following conditions: enzyme loading 10 FPU/g-substrate, substrate 5.5 %, temperature 50 °C, and pH 4.5. The hydrolysis yield obtained using the enzymes from the fungal consortium was equivalent to that obtained using a mixture of commercial enzymes Celluclast and Novozyme β-glucosidase. Addition of up to 500 ppm of heavy metal ions (As, Cu, Fe, Mn, Ni, Pb, and Zn) during saccharification did not significantly affect the saccharification yield. Thus, the biomass grown for phytoremediation of heavy metals can be used for the production of reducing sugars followed by ethanol fermentation.

  14. Optimisation of Environmental Conditions for Enhanced Production of Fungal Exopectinase Using Agro-industrial Wastes.

    Science.gov (United States)

    Akbar, Sabika; Prasuna, R Gyana; Theja, B; Chakradhar, Y M S

    2014-07-01

    Management of household solid waste and agro industrial residues generated from various sources is a serious problem due to huge ever increasing population and pollution. Application of these worthless agro waste materials to generate a commercially valuable product, pectinase enzyme, using locally isolated fungal strain, Aspergillus flavipes, was the main motive of this study. Physiological characterisation and enzyme profile determination were done along with formulation of production media. Fruit skins, rags were used as C source and oil cakes were used for N source. Various combinations of these C and N sources were applied for revised production of pectinase enzyme compared to YEP basal media (29 U/ml). A huge increase in pectinase production of 40 U/ml was obtained with Citrus peel - Sesame oil cake (CS) media. The enzyme had its maximum activity at 500C, 4.5 pH. This was achieved at 45 min in 1.5% substrate concentration.

  15. Controlled enzyme catalyzed heteropolysaccharide degradation:Xylans

    OpenAIRE

    Rasmussen, Louise Enggaard; Meyer, Anne S.

    2011-01-01

    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 monocompo...

  16. The ENZYME data bank.

    Science.gov (United States)

    Bairoch, A

    1994-09-01

    The ENZYME data bank is a repository of information relative to the nomenclature of enzymes. It is primarily based on the recommendations of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB) and it contains the following data for each type of characterized enzyme for which an EC (Enzyme Commission) number has been provided: EC number Recommended name Alternative names (if any) Catalytic activity Cofactors (if any) Pointers to the SWISS-PROT protein sequence entrie(s) that correspond to the enzyme (if any) Pointers to human disease(s) associated with a deficiency of the enzyme (if any).

  17. Air Contamination With Fungals In Museum

    Science.gov (United States)

    Scarlat, Iuliana; Haiducu, Maria; Stepa, Raluca

    2015-07-01

    The aim of the studies was to determine the level and kind of fungal contamination of air in museum, deposits patrimony, restoration and conservation laboratories and their effects on health of workers. Microbiological air purity was measured with a SAS-100 Surface Air System impactor. The fungal contamination was observed in all 54 rooms where we made determinations. The highest levels of fungal were recorded at rooms with hygroscopic patrimony objects, eg carpets, chairs, upholstered chairs, books etc. The most species identified included under common allergens: Aspergillus, Penicillium, and Mucor. There fungal species belonging to the genus identified in this study, can trigger serious diseases museum workers, such as for example Aspergillus fumigatus, known allergies and toxic effects that may occur. In some places of the museum, occupational exposure limit values to fungi present in the air in the work environment, recommended by the specialized literature, have been overcome.

  18. Fungal keratitis - improving diagnostics by confocal microscopy

    DEFF Research Database (Denmark)

    Nielsen, Esben; Heegaard, S; Prause, J U

    2013-01-01

    Purpose: Introducing a simple image grading system to support the interpretation of in vivo confocal microscopy (IVCM) images in filamentous fungal keratitis. Setting: Clinical and confocal studies took place at the Department of Ophthalmology, Aarhus University Hospital, Denmark. Histopathological...... analysis was performed at the Eye Pathology Institute, Department of Neuroscience and Pharmacology, University of Copenhagen, Denmark. Methods: A recent series of consecutive patients with filamentous fungal keratitis is presented to demonstrate the results from in-house IVCM. Based upon our experience...... with IVCM and previously published images, we composed a grading system for interpreting IVCM images of filamentous fungal keratitis. Results: A recent case series of filamentous fungal keratitis from 2011 to 2012 was examined. There were 3 male and 3 female patients. Mean age was 44.5 years (range 12...

  19. Stem Cell Transplant Patients and Fungal Infections

    Science.gov (United States)

    ... includes places like chicken coops and caves. Wear gloves when handling materials such as soil, moss, or ... Cell Transplant Recipients. MMWR 2000;49:1-128. Top of Page Related Links Fungal Meningitis National Center ...

  20. Fungal rhino sinusitisin in tehran, iran

    NARCIS (Netherlands)

    Nazeri, M.; Hashemi, S.J.; Ardehali, M.; Rezaei, S.; Seyedmousavi, S.; Zareei, M.; Hosseinjani, E.

    2015-01-01

    BACKGROUND: Fungal rhino sinusitis (FRS) is an important infection of para nasal sinuses, which encompasses two main categories; invasive and noninvasive forms according to histopathological findings. Aspergillus spp are the most common species isolated from noninvasive form, while Mucorales are

  1. Foreword: Special issue on fungal grapevine diseases

    Science.gov (United States)

    An impressively large proportion of fungicides applied in European, North American and Australian agriculture has been used to manage grapevine powdery mildew (Erysiphe necator), grapevine downy mildew (Plasmopara viticola), and botrytis bunch rot (Botrytis cinerea). These fungal and oomycetous plan...

  2. Marine fungal biotechnology: An ecological perspective

    Digital Repository Service at National Institute of Oceanography (India)

    Raghukumar, C.

    microscopy and immunocytochemical labelling of β-1,4-N- acetyl-D-glucosamine residues of chitin walls, a fungal signature confirmed its presence (Maldonado et al., 2005). Interestingly these yeasts are reported to be maternally transmitted from the soma...

  3. Zoosporic fungal parasites of marine biota

    Digital Repository Service at National Institute of Oceanography (India)

    RaghuKumar, C.

    laboratory media. In such instances, a detailed and careful examination of the disease symptoms and the endobiotic fungal parasites is to be recorded. Maintaining dual culture of the healthy and infected host also helps to fulfill these postulates partially....

  4. Contaminations Occurring in Fungal PCR Assays

    Science.gov (United States)

    Loeffler, Juergen; Hebart, Holger; Bialek, Ralf; Hagmeyer, Lars; Schmidt, Diethard; Serey, Francois-Prâseth; Hartmann, Matthias; Eucker, Jan; Einsele, Hermann

    1999-01-01

    Successful in vitro amplification of fungal DNA in clinical specimens has been reported recently. In a collaboration among five European centers, the frequency and risk of contamination due to airborne spore inoculation or carryover contamination in fungal PCR were analyzed. The identities of all contaminants were specified by cycle sequencing and GenBank analysis. Twelve of 150 PCR assays that together included over 2,800 samples were found to be contaminated (3.3% of the negative controls were contaminated during the DNA extraction, and 4.7% of the PCR mixtures were contaminated during the amplification process). Contaminants were specified as Aspergillus fumigatus, Saccharomyces cerevisiae, and Acremonium spp. Further analysis showed that commercially available products like zymolyase powder or 10× PCR buffer may contain fungal DNA. In conclusion, the risk of contamination is not higher in fungal PCR assays than in other diagnostic PCR-based assays if general precautions are taken. PMID:10074553

  5. Spectral Characterization of Fungal Metabolites in Aqueous Medium with Humus Substances

    Directory of Open Access Journals (Sweden)

    D. A. Khundzhua

    2013-01-01

    Full Text Available The work is targeted to confirm participation of microscopic fungi in transformation of humus substances in aquatic environments. The research is focused on the spectroscopic study of the collection of fungal strains with different pigmentation of mycelium. Spectral properties of fungal metabolites were measured and compared to that of natural aquatic nonliving organic matter and commercial humus substances in aqueous solutions. The experiments revealed that the effect of microscopic fungi growing in the culture medium with added humate appeared as changes in the humic-type fluorescence: its characteristics became more similar to that of nonliving organic matter in natural waters than to original humate preparation. The experiments demonstrated degradation of coal-originated humate due to microbial activity into compounds of smaller molecular size and increased heterogeneity. We resume that transformation of humus substances by fungal cultures can be monitored and characterized using spectral measurements.

  6. Fungal Endophthalmitis Associated with Compounded Products

    OpenAIRE

    Mikosz, Christina A.; Smith, Rachel M.; Kim, Moon; Tyson, Clara; Lee, Ellen H.; Adams, Eleanor; Straif-Bourgeois, Susanne; Sowadsky, Rick; Arroyo, Shannon; Grant-Greene, Yoran; Duran, Julie; Vasquez, Yvonne; Robinson, Byron F.; Harris, Julie R.; Lockhart, Shawn R.

    2014-01-01

    Fungal endophthalmitis is a rare but serious infection. In March 2012, several cases of probable and laboratory-confirmed fungal endophthalmitis occurring after invasive ocular procedures were reported nationwide. We identified 47 cases in 9 states: 21 patients had been exposed to the intraocular dye Brilliant Blue G (BBG) during retinal surgery, and the other 26 had received an intravitreal injection containing triamcinolone acetonide. Both drugs were produced by Franck’s Compounding Lab (Oc...

  7. Nail Histomycology, Onychochromobiology, and Fungal Thigmatropism

    OpenAIRE

    Gérald E. Piérard; Sébastien L. Piérard

    2016-01-01

    Background: Thigmotropism is a biologic feature coping with the directional growth of cells following topographical guidance cues. This mechanism is involved in the invasive phase of pathogen and opportunistic fungi. It was shown experimentally with fungal hyphae of both dermatophytes and nondermatophyte molds, as well as with the mycelial phase of the dimorphic yeast Candida albicans. Objective: To revisit histomycology in onychomycoses of a diversity of fungal origins. Method: Histopatholog...

  8. Allergic Fungal Rhinosinusitis - Outcomes of Multimodality Treatment

    OpenAIRE

    Baisakhi Bakat; Subhendu Chowdhury; Amitabha Roy Chowdhury; Soumitra Ghosh; Barin Kumar Roychaudhuri

    2013-01-01

    Abstract Objective : To evaluate the treatment outcomes of multimodality therapy for allergic fungal rhinosinusitis. Study Design: Prospective Observational Study. Materials&methods : This study was carried out in the department of ENT & Head neck Surgery, Vivekananda Institute of Medical Sciences, Ramakrishna Mission Seva Pratishthan, Kolkata from January 2010 to July 2011. During this study period of one & half years, 20 subjects having Allergic Fungal Sinusitis were selected fr...

  9. Biological roles of fungal carotenoids.

    Science.gov (United States)

    Avalos, Javier; Carmen Limón, M

    2015-08-01

    Carotenoids are terpenoid pigments widespread in nature, produced by bacteria, fungi, algae and plants. They are also found in animals, which usually obtain them through the diet. Carotenoids in plants provide striking yellow, orange or red colors to fruits and flowers, and play important metabolic and physiological functions, especially relevant in photosynthesis. Their functions are less clear in non-photosynthetic microorganisms. Different fungi produce diverse carotenoids, but the mutants unable to produce them do not exhibit phenotypic alterations in the laboratory, apart of lack of pigmentation. This review summarizes the current knowledge on the functional basis for carotenoid production in fungi. Different lines of evidence support a protective role of carotenoids against oxidative stress and exposure to visible light or UV irradiation. In addition, the carotenoids are intermediary products in the biosynthesis of physiologically active apocarotenoids or derived compounds. This is the case of retinal, obtained from the symmetrical oxidative cleavage of β-carotene. Retinal is the light-absorbing prosthetic group of the rhodopsins, membrane-bound photoreceptors present also in many fungal species. In Mucorales, β-carotene is an intermediary in the synthesis of trisporoids, apocarotenoid derivatives that include the sexual hormones the trisporic acids, and they are also presumably used in the synthesis of sporopollenin polymers. In conclusion, fungi have adapted their ability to produce carotenoids for different non-essential functions, related with stress tolerance or with the synthesis of physiologically active by-products.

  10. Burden of fungal infections in Senegal.

    Science.gov (United States)

    Badiane, Aida S; Ndiaye, Daouda; Denning, David W

    2015-10-01

    Senegal has a high rate of tuberculosis and a low HIV seropositivity rate and aspergilloma, life-threatening fungal infections, dermatophytosis and mycetoma have been reported in this study. All published epidemiology papers reporting fungal infection rates from Senegal were identified. Where no data existed, we used specific populations at risk and fungal infection frequencies in each to estimate national incidence or prevalence. The results show that tinea capitis is common being found in 25% of children, ~1.5 million. About 191,000 Senegalese women get recurrent vaginal thrush, ≥4 times annually. We estimate 685 incident cases of chronic pulmonary aspergillosis (CPA) following TB and prevalence of 2160 cases. Asthma prevalence in adults varies from 3.2% to 8.2% (mean 5%); 9976 adults have allergic bronchopulmonary aspergillosis (ABPA) and 13,168 have severe asthma with fungal sensitisation (SAFS). Of the 59,000 estimated HIV-positive patients, 366 develop cryptococcal meningitis; 1149 develop Pneumocystis pneumonia and 1946 develop oesophageal candidiasis, in which oral candidiasis (53%) and dermatophytosis (16%) are common. Since 2008-2010, 113 cases of mycetoma were diagnosed. In conclusion, we estimate that 1,743,507 (12.5%) people in Senegal suffer from a fungal infection, excluding oral candidiasis, fungal keratitis, invasive candidiasis or aspergillosis. Diagnostic and treatment deficiencies should be rectified to allow epidemiological studies. © 2015 Blackwell Verlag GmbH.

  11. Fungal keratitis - improving diagnostics by confocal microscopy.

    Science.gov (United States)

    Nielsen, E; Heegaard, S; Prause, J U; Ivarsen, A; Mortensen, K L; Hjortdal, J

    2013-09-01

    Introducing a simple image grading system to support the interpretation of in vivo confocal microscopy (IVCM) images in filamentous fungal keratitis. Clinical and confocal studies took place at the Department of Ophthalmology, Aarhus University Hospital, Denmark. Histopathological analysis was performed at the Eye Pathology Institute, Department of Neuroscience and Pharmacology, University of Copenhagen, Denmark. A recent series of consecutive patients with filamentous fungal keratitis is presented to demonstrate the results from in-house IVCM. Based upon our experience with IVCM and previously published images, we composed a grading system for interpreting IVCM images of filamentous fungal keratitis. A recent case series of filamentous fungal keratitis from 2011 to 2012 was examined. There were 3 male and 3 female patients. Mean age was 44.5 years (range 12-69), 6 out of 17 (35%) cultures were positive and a total of 6/7 (86%) IVCM scans were positive. Three different categories of IVCM results for the grading of diagnostic certainty were formed. IVCM is a valuable tool for diagnosing filamentous fungal keratitis. In order to improve the reliability of IVCM, we suggest implementing a simple and clinically applicable grading system for aiding the interpretation of IVCM images of filamentous fungal keratitis.

  12. Fungal Keratitis - Improving Diagnostics by Confocal Microscopy

    Directory of Open Access Journals (Sweden)

    Esben Nielsen

    2013-12-01

    Full Text Available Purpose: Introducing a simple image grading system to support the interpretation of in vivo confocal microscopy (IVCM images in filamentous fungal keratitis. Setting: Clinical and confocal studies took place at the Department of Ophthalmology, Aarhus University Hospital, Denmark. Histopathological analysis was performed at the Eye Pathology Institute, Department of Neuroscience and Pharmacology, University of Copenhagen, Denmark. Methods: A recent series of consecutive patients with filamentous fungal keratitis is presented to demonstrate the results from in-house IVCM. Based upon our experience with IVCM and previously published images, we composed a grading system for interpreting IVCM images of filamentous fungal keratitis. Results: A recent case series of filamentous fungal keratitis from 2011 to 2012 was examined. There were 3 male and 3 female patients. Mean age was 44.5 years (range 12-69, 6 out of 17 (35% cultures were positive and a total of 6/7 (86% IVCM scans were positive. Three different categories of IVCM results for the grading of diagnostic certainty were formed. Conclusion: IVCM is a valuable tool for diagnosing filamentous fungal keratitis. In order to improve the reliability of IVCM, we suggest implementing a simple and clinically applicable grading system for aiding the interpretation of IVCM images of filamentous fungal keratitis.

  13. Structure and biological functions of fungal cerebrosides

    Directory of Open Access Journals (Sweden)

    Barreto-Bergter Eliana

    2004-01-01

    Full Text Available Ceramide monohexosides (CMHs, cerebrosides are glycosphingolipids composed of a hydrophobic ceramide linked to one sugar unit. In fungal cells, CMHs are very conserved molecules consisting of a ceramide moiety containing 9-methyl-4,8-sphingadienine in amidic linkage to 2-hydroxyoctadecanoic or 2-hydroxyhexadecanoic acids, and a carbohydrate portion consisting of one residue of glucose or galactose. 9-Methyl 4,8-sphingadienine-containing ceramides are usually glycosylated to form fungal cerebrosides, but the recent description of a ceramide dihexoside (CDH presenting phytosphingosine in Magnaporthe grisea suggests the existence of alternative pathways of ceramide glycosylation in fungal cells. Along with their unique structural characteristics, fungal CMHs have a peculiar subcellular distribution and striking biological properties. In Pseudallescheria boydii, Candida albicans, Cryptococcus neoformans, Aspergillus nidulans, A. fumigatus, and Schizophyllum commune, CMHs are apparently involved in morphological transitions and fungal growth. The elucidation of structural and functional aspects of fungal cerebrosides may therefore contribute to the design of new antifungal agents inhibiting growth and differentiation of pathogenic species.

  14. Transcriptional Profiling of a Yeast Colony Provides New Insight into the Heterogeneity of Multicellular Fungal Communities

    Science.gov (United States)

    Traven, Ana; Jänicke, Amrei; Harrison, Paul; Swaminathan, Angavai; Seemann, Torsten; Beilharz, Traude H.

    2012-01-01

    Understanding multicellular fungal structures is important for designing better strategies against human fungal pathogens. For example, the ability to form multicellular biofilms is a key virulence property of the yeast Candida albicans. C. albicans biofilms form on indwelling medical devices and are drug resistant, causing serious infections in hospital settings. Multicellular fungal communities are heterogeneous, consisting of cells experiencing different environments. Heterogeneity is likely important for the phenotypic characteristics of communities, yet it is poorly understood. Here we used colonies of the yeast Saccharomyces cerevisiae as a model fungal multicellular structure. We fractionated the outside colony layers from the cells in the center by FACS, using a Cit1-GFP marker expressed exclusively on the outside. Transcriptomics analysis of the two subpopulations revealed that the outside colony layers are actively growing by fermentative metabolism, while the cells residing on the inside are in a resting state and experience changes to mitochondrial activity. Our data shows several parallels with C. albicans biofilms providing insight into the contributions of heterogeneity to biofilm phenotypes. Hallmarks of C. albicans biofilms – the expression of ribosome and translation functions and activation of glycolysis and ergosterol biosynthesis occur on the outside of colonies, while expression of genes associates with sulfur assimilation is observed in the colony center. Cell wall restructuring occurs in biofilms, and cell wall functions are enriched in both fractions: the outside cells display enrichment of cell wall biosynthesis enzymes and cell wall proteins, while the inside cells express cell wall degrading enzymes. Our study also suggests that noncoding transcription and posttranscriptional mRNA regulation play important roles during growth of yeast in colonies, setting the scene for investigating these pathways in the development of multicellular

  15. Transcriptional profiling of a yeast colony provides new insight into the heterogeneity of multicellular fungal communities.

    Directory of Open Access Journals (Sweden)

    Ana Traven

    Full Text Available Understanding multicellular fungal structures is important for designing better strategies against human fungal pathogens. For example, the ability to form multicellular biofilms is a key virulence property of the yeast Candida albicans. C. albicans biofilms form on indwelling medical devices and are drug resistant, causing serious infections in hospital settings. Multicellular fungal communities are heterogeneous, consisting of cells experiencing different environments. Heterogeneity is likely important for the phenotypic characteristics of communities, yet it is poorly understood. Here we used colonies of the yeast Saccharomyces cerevisiae as a model fungal multicellular structure. We fractionated the outside colony layers from the cells in the center by FACS, using a Cit1-GFP marker expressed exclusively on the outside. Transcriptomics analysis of the two subpopulations revealed that the outside colony layers are actively growing by fermentative metabolism, while the cells residing on the inside are in a resting state and experience changes to mitochondrial activity. Our data shows several parallels with C. albicans biofilms providing insight into the contributions of heterogeneity to biofilm phenotypes. Hallmarks of C. albicans biofilms - the expression of ribosome and translation functions and activation of glycolysis and ergosterol biosynthesis occur on the outside of colonies, while expression of genes associates with sulfur assimilation is observed in the colony center. Cell wall restructuring occurs in biofilms, and cell wall functions are enriched in both fractions: the outside cells display enrichment of cell wall biosynthesis enzymes and cell wall proteins, while the inside cells express cell wall degrading enzymes. Our study also suggests that noncoding transcription and posttranscriptional mRNA regulation play important roles during growth of yeast in colonies, setting the scene for investigating these pathways in the development

  16. Crystallization of a fungal lytic polysaccharide monooxygenase expressed from glycoengineered Pichia pastoris for X-ray and neutron diffraction.

    Science.gov (United States)

    O'Dell, William B; Swartz, Paul D; Weiss, Kevin L; Meilleur, Flora

    2017-02-01

    Lytic polysaccharide monooxygenases (LPMOs) are carbohydrate-disrupting enzymes secreted by bacteria and fungi that break glycosidic bonds via an oxidative mechanism. Fungal LPMOs typically act on cellulose and can enhance the efficiency of cellulose-hydrolyzing enzymes that release soluble sugars for bioethanol production or other industrial uses. The enzyme PMO-2 from Neurospora crassa (NcPMO-2) was heterologously expressed in Pichia pastoris to facilitate crystallographic studies of the fungal LPMO mechanism. Diffraction resolution and crystal morphology were improved by expressing NcPMO-2 from a glycoengineered strain of P. pastoris and by the use of crystal seeding methods, respectively. These improvements resulted in high-resolution (1.20 Å) X-ray diffraction data collection at 100 K and the production of a large NcPMO-2 crystal suitable for room-temperature neutron diffraction data collection to 2.12 Å resolution.

  17. Delignification of Wood Chips and Pulps by Using Natural and Synthetic Porphyrins: Models of Fungal Decay †

    Science.gov (United States)

    Paszczyński, Andrzej; Crawford, Ronald L.; Blanchette, Robert A.

    1988-01-01

    Kraft pulps, prepared from softwoods, and small chips of birch wood were treated with heme and tert-butyl hydroperoxide in aqueous solutions at reflux temperature. Analyses of treated pulps showed decreases in kappa number (a measure of lignin content) from about 36 to less than 2, with concomitant increases in brightness (80% increase in the better samples). Analyses of treated wood chips revealed selective delignification and removal of hemicelluloses. After 48 h of treatment, lignin losses from the wood chips approached 40%, and xylose/mannose (hemicellulose) losses approached 70%, while glucose (cellulose) losses were less than 10%. Examination of delignified chips by transmission electron microscopy showed that the removal of lignin occurred in a manner virtually indistinguishable from that seen after decay by white rot fungi. Various metalloporphyrins, which act as biomimetic catalysts, were compared to horseradish peroxidase and fungal manganese peroxidase in their abilities to oxidize syringaldazine in an organic solvent, dioxane. The metalloporphyrins and peroxidases behaved similarly, and it appeared that the activities of the peroxidases resulted from the extraction of heme into the organic phase, rather than from the activities of the enzymes themselves. We concluded that heme-tert-butyl hydroperoxide systems in the absence of a protein carrier mimic the decay of lignified tissues by white rot fungi. Images PMID:16347540

  18. ENZYME RESISTANCE OF GENETICALLY MODIFIED STARCH POTATOES

    Directory of Open Access Journals (Sweden)

    A. Sh. Mannapova

    2015-01-01

    Full Text Available Here in this article the justification of expediency of enzyme resistant starch use in therapeutic food products is presented . Enzyme resistant starch is capable to resist to enzymatic hydrolysis in a small intestine of a person, has a low glycemic index, leads to decrease of postprandial concentration of glucose, cholesterol, triglycerides in blood and insulin reaction, to improvement of sensitivity of all organism to insulin, to increase in sense of fulness and to reduction of adjournment of fats. Resistant starch makes bifidogenшс impact on microflora of a intestine of the person, leads to increase of a quantity of lactobacillus and bifidobacterium and to increased production of butyric acid in a large intestine. In this regard the enzyme resistant starch is an important component in food for prevention and curing of human diseases such as diabetes, obesity, colitis, a cancer of large and direct intestine. One method is specified by authors for imitation of starch digestion in a human body. This method is based on the definition of an enzyme resistance of starch in vitro by its hydrolysis to glucose with application of a glucoamylase and digestive enzyme preparation Pancreatin. This method is used in researches of an enzyme resistance of starch, of genetically modified potato, high amylose corn starch Hi-Maize 1043 and HYLON VII (National Starch Food Innovation, USA, amylopectin and amylose. It is shown that the enzyme resistance of the starch emitted from genetically modified potatoes conforms to the enzyme resistance of the high amylose corn starch “Hi-Maize 1043 and HYLON VII starch”, (National Starch Food Innovation, the USA relating to the II type of enzyme resistant starch. It is established that amylopectin doesn't have the enzyme resistant properties. The results of researches are presented. They allow us to make the following conclusion: amylose in comparison with amylopectin possesses higher enzyme resistance and gives to

  19. Enzyme inhibition by iminosugars

    DEFF Research Database (Denmark)

    López, Óscar; Qing, Feng-Ling; Pedersen, Christian Marcus

    2013-01-01

    Imino- and azasugar glycosidase inhibitors display pH dependant inhibition reflecting that both the inhibitor and the enzyme active site have groups that change protonation state with pH. With the enzyme having two acidic groups and the inhibitor one basic group, enzyme-inhibitor complexes...

  20. Fungal-host diversity among mycoheterotrophic plants increases proportionally to their fungal-host overlap.

    Science.gov (United States)

    Gomes, Sofia I F; Merckx, Vincent S F T; Saavedra, Serguei

    2017-05-01

    The vast majority of plants obtain an important proportion of vital resources from soil through mycorrhizal fungi. Generally, this happens in exchange of photosynthetically fixed carbon, but occasionally the interaction is mycoheterotrophic, and plants obtain carbon from mycorrhizal fungi. This process results in an antagonistic interaction between mycoheterotrophic plants and their fungal hosts. Importantly, the fungal-host diversity available for plants is restricted as mycoheterotrophic interactions often involve narrow lineages of fungal hosts. Unfortunately, little is known whether fungal-host diversity may be additionally modulated by plant-plant interactions through shared hosts. Yet, this may have important implications for plant competition and coexistence. Here, we use DNA sequencing data to investigate the interaction patterns between mycoheterotrophic plants and arbuscular mycorrhizal fungi. We find no phylogenetic signal on the number of fungal hosts nor on the fungal hosts shared among mycoheterotrophic plants. However, we observe a potential trend toward increased phylogenetic diversity of fungal hosts among mycoheterotrophic plants with increasing overlap in their fungal hosts. While these patterns remain for groups of plants regardless of location, we do find higher levels of overlap and diversity among plants from the same location. These findings suggest that species coexistence cannot be fully understood without attention to the two sides of ecological interactions.

  1. Modification of plasma membrane electron transport in cultured rose cells by UV-C radiation and fungal elicitor

    International Nuclear Information System (INIS)

    Murphy, T.M.; Auh, C.K.; Schorr, R.; Grobe, C.

    1991-01-01

    Previous experiments have shown that treatments of suspension-cultured cells of Rosa damascena Mill. with UV radiation or with fungal elicitors stimulates the synthesis of H 2 O 2 by the cells. To test the hypothesis that this synthesis involves reduction of O 2 at the plasma membrane and to identify the mechanism of the reduction, we have determined the effects of UV and elicitor on redox reactions associated with the plasma membrane. Elicitor prepared from cell walls of Phytophthora sp. (14 μg solids/ml) inhibited the reduction of ferricyanide by intact cells by 98%; UV-C (primarily 254 nm, up to 19,500 J/m 2 ) inhibited this reduction by 40%. Neither treatment inhibited the reduction of Fe(III)-EDTA by intact cells. Intact cells oxidized NADH in the absence of external oxidizing agent, and the rate of oxidation was increased by UV and elicitor. Cells that were poisoned with arsenite and CCCP catalyzed the reduction of Fe(III)-EDTA in the presence of external NADH, and this ability was slightly stimulated by UV and elicitor. UV irradiation (6,480 J/m 2 ) of cells resulted in a 27% inhibition of the specific activity of NADH-ferricyanide oxidoreductase in plasma membrane isolated from those cells. Elicitor treatment of cells for at least 90 min resulted in a 50% inhibition of the enzyme's specific activity in isolated plasma membrane; this inhibition was reversed by addition of Triton-X100 in the assay mixture. The results suggest that UV and elicitor alter the flow of electrons in the plasma membrane, reversibly inhibiting NADH-cytochrome b reductase, the putative key enzyme in the pathway of ferricyanide reduction, and stimulating (or at least not inhibiting) the pathway of Fe(III)-EDTA reduction

  2. Chitinolytic enzymes from Clostridium aminovalericum: activity screening and purification.

    Science.gov (United States)

    Simůnek, J; Tishchenko, G; Rozhetsky, K; Bartonová, H; Kopecný, J; Hodrová, B

    2004-01-01

    A strain isolated from the feces of takin was identified as Clostridium aminovalericum. In response to various types of chitin used as growth substrates, the bacterium produced a complete array of chitinolytic enzymes: chitinase ('endochitinase'), exochitinase, beta-N-acetylglucosaminidase, chitosanase and chitin deacetylase. The highest activities of chitinase (536 pkat/mL) and exochitinase (747 pkat/mL) were induced by colloidal chitin. Fungal chitin also induced high levels of these enzymes (463 pkat/mL and 502 pkat/mL, respectively). Crab shell chitin was the best inducer of chitosanase activity (232 pkat/mL). The chitinolytic enzymes of this strain were separated from culture filtrate by ion-exchange chromatography on the carboxylic sorbent Polygran 27. At pH 4.5, some isoforms of the chitinolytic enzymes (30% of total enzyme activity) did not bind to Polygran 27. The enzymes were eluted under a stepwise pH gradient (pH 5-8) in 0.1 mol/L phosphate buffer. At merely acidic pH (4.5-5.5), the adsorbed enzymes were co-eluted. However, at pH close to neutral values, the peaks of highly purified isoforms of exochitinases and chitinases were isolated. The protein and enzyme recovery reached 90%.

  3. Bisphenol A degradation in water by ligninolytic enzymes.

    Science.gov (United States)

    Gassara, Fatma; Brar, Satinder K; Verma, M; Tyagi, R D

    2013-08-01

    Many endocrine disruptor compounds, such as bisphenol A (BPA) are used today and released into the environment at low doses but they are barely degraded in wastewater treatment plants. One of the potential alternatives to effectively degrade endocrine disruptor compounds is based on the use of the oxidative action of extracellular fungal enzymes. The aim of this work is to study the ability of free and encapsulated enzymes (manganese peroxidase, lignin peroxidase and laccase) to degrade BPA. Higher degradation of BPA (90%) by ligninolytic enzymes encapsulated on polyacrylamide hydrogel and pectin after 8h was obtained. The degradation of BPA while using the free enzyme (26%) was lower than the value obtained with encapsulated enzymes. The presence of pectin in the formulation significantly (p>0.05) enhanced the activity of enzymes. Kinetics of BPA degradation showed an increase in Vm, while Km remained constant when enzymes were encapsulated. Hence, encapsulation protected the enzymes from non-competitive inhibition. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. CRISPR-Cas9, the new kid on the block of fungal molecular biology.

    Science.gov (United States)

    Krappmann, Sven

    2017-01-01

    Research on fungal pathogens with the aim to identify virulence determinants strictly relies on the generation of defined, recombinant strains, a task that is executed by means of a sophisticated molecular biology toolbox. Recent developments in fungal genome engineering have opened a new frontier by implementing the CRISPR-Cas9 technology, based on expression of the Cas9 endonuclease that is loaded by a single guiding RNA (sgRNA) molecule to target a defined site in the recipient genome. This novel approach has been adapted successfully to engineer fungal genomes, among them the one of the human-pathogenic mould Aspergillus fumigatus Implementation of the required components was achieved by various means that differ with respect to expression of the Cas9 enzyme and sgRNA delivery. Validation of CRISPR-Cas9-mediated mutagenesis could be executed by targeting selected candidate genes of A. fumigatus to provide a promising perspective for screening and multiplexing approaches to scrutinize the virulome of this opportunistic fungal pathogen in a comprehensive manner, such as by analyzing genetic polymorphisms or the function of gene families. © The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  5. Bioremediation of MGP soils with mixed fungal and bacterial cultures

    International Nuclear Information System (INIS)

    Lee, C.J.B.; Fletcher, M.A.; Avila, O.I.; Munnecke, D.M.; Callanan, J.; Yunker, S.

    1995-01-01

    This culture selection study examines the degradation of polycyclic automatic hydrocarbon (PAH) by a number of brown- and white-rot fungi and bacterial cultures for the treatment of coal tar wastes. Cultures were screened for naphthalene degradation in shake flasks, and selected organisms were then examined for their ability to degrade a mixture of PAHs in aqueous culture. PAH degradation in the presence of the surfactant, TWEEN 80, was examined for some cultures. Many of the organisms were observed to be resistant to greater than 10 mg/L free cyanide. Solid substrate growth conditions were optimized for the selected fungal cultures in preparation for manufactured gas plant (MGP) soil microcosm experiments. The fungi generally produced more biomass under conditions of acidic to neutral pH, incubation at 30 C with 90% moisture saturation, and with granulated corncobs or alfalfa pellets supplied as a lignocellulosic substrate. Of the cultures screened, nine fungal cultures were selected based on their ability to degrade at least 40% of naphthalene, fluorene, or benzo(a)pyrene in 2 weeks or less. A bacterial culture capable of degrading 30 mg/L of naphthalene in 1 week was also selected, and the cultures were examined further in PAH-degradation studies in contaminated soils

  6. Proteomics of survival structures of fungal pathogens.

    Science.gov (United States)

    Loginov, Dmitry; Šebela, Marek

    2016-09-25

    Fungal pathogens are causal agents of numerous human, animal, and plant diseases. They employ various infection modes to overcome host defense systems. Infection mechanisms of different fungi have been subjected to many comprehensive studies. These investigations have been facilitated by the development of various '-omics' techniques, and proteomics has one of the leading roles in this regard. Fungal conidia and sclerotia could be considered the most important structures for pathogenesis as their germination is one of the first steps towards a host infection. They represent interesting objects for proteomic studies because of the presence of unique proteins with unexplored biotechnological potential required for pathogen viability, development and the subsequent host infection. Proteomic peculiarities of survival structures of different fungi, including those of biotechnological significance (e.g., Asperillus fumigatus, A. nidulans, Metarhizium anisopliae), in a dormant state, as well as changes in the protein production during early stages of fungal development are the subjects of the present review. We focused on biological aspects of proteomic studies of fungal survival structures rather than on an evaluation of proteomic approaches. For that reason, proteins that have been identified in this context are discussed from the point of view of their involvement in different biological processes and possible functions assigned to them. This is the first review paper summarizing recent advances in proteomics of fungal survival structures. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Fungal endophytes: modifiers of plant disease.

    Science.gov (United States)

    Busby, Posy E; Ridout, Mary; Newcombe, George

    2016-04-01

    Many recent studies have demonstrated that non-pathogenic fungi within plant microbiomes, i.e., endophytes ("endo" = within, "phyte" = plant), can significantly modify the expression of host plant disease. The rapid pace of advancement in endophyte ecology warrants a pause to synthesize our understanding of endophyte disease modification and to discuss future research directions. We reviewed recent literature on fungal endophyte disease modification, and here report on several emergent themes: (1) Fungal endophyte effects on plant disease span the full spectrum from pathogen antagonism to pathogen facilitation, with pathogen antagonism most commonly reported. (2) Agricultural plant pathosystems are the focus of research on endophyte disease modification. (3) A taxonomically diverse group of fungal endophytes can influence plant disease severity. And (4) Fungal endophyte effects on plant disease severity are context-dependent. Our review highlights the importance of fungal endophytes for plant disease across a broad range of plant pathosystems, yet simultaneously reveals that complexity within plant microbiomes presents a significant challenge to disentangling the biotic environmental factors affecting plant disease severity. Manipulative studies integrating eco-evolutionary approaches with emerging molecular tools will be poised to elucidate the functional importance of endophytes in natural plant pathosystems that are fundamental to biodiversity and conservation.

  8. Fueling the Future with Fungal Genomes

    Energy Technology Data Exchange (ETDEWEB)

    Grigoriev, Igor V.

    2014-10-27

    Genomes of fungi relevant to energy and environment are in focus of the JGI Fungal Genomic Program. One of its projects, the Genomics Encyclopedia of Fungi, targets fungi related to plant health (symbionts and pathogens) and biorefinery processes (cellulose degradation and sugar fermentation) by means of genome sequencing and analysis. New chapters of the Encyclopedia can be opened with user proposals to the JGI Community Science Program (CSP). Another JGI project, the 1000 fungal genomes, explores fungal diversity on genome level at scale and is open for users to nominate new species for sequencing. Over 400 fungal genomes have been sequenced by JGI to date and released through MycoCosm (www.jgi.doe.gov/fungi), a fungal web-portal, which integrates sequence and functional data with genome analysis tools for user community. Sequence analysis supported by functional genomics will lead to developing parts list for complex systems ranging from ecosystems of biofuel crops to biorefineries. Recent examples of such ‘parts’ suggested by comparative genomics and functional analysis in these areas are presented here.

  9. Fungal Genomics for Energy and Environment

    Energy Technology Data Exchange (ETDEWEB)

    Grigoriev, Igor V.

    2013-03-11

    Genomes of fungi relevant to energy and environment are in focus of the Fungal Genomic Program at the US Department of Energy Joint Genome Institute (JGI). One of its projects, the Genomics Encyclopedia of Fungi, targets fungi related to plant health (symbionts, pathogens, and biocontrol agents) and biorefinery processes (cellulose degradation, sugar fermentation, industrial hosts) by means of genome sequencing and analysis. New chapters of the Encyclopedia can be opened with user proposals to the JGI Community Sequencing Program (CSP). Another JGI project, the 1000 fungal genomes, explores fungal diversity on genome level at scale and is open for users to nominate new species for sequencing. Over 200 fungal genomes have been sequenced by JGI to date and released through MycoCosm (www.jgi.doe.gov/fungi), a fungal web-portal, which integrates sequence and functional data with genome analysis tools for user community. Sequence analysis supported by functional genomics leads to developing parts list for complex systems ranging from ecosystems of biofuel crops to biorefineries. Recent examples of such parts suggested by comparative genomics and functional analysis in these areas are presented here.

  10. Fungal endophytes for sustainable crop production.

    Science.gov (United States)

    Lugtenberg, Ben J J; Caradus, John R; Johnson, Linda J

    2016-12-01

    This minireview highlights the importance of endophytic fungi for sustainable agriculture and horticulture production. Fungal endophytes play a key role in habitat adaptation of plants resulting in improved plant performance and plant protection against biotic and abiotic stresses. They encode a vast variety of novel secondary metabolites including volatile organic compounds. In addition to protecting plants against pathogens and pests, selected fungal endophytes have been used to remove animal toxicities associated with fungal endophytes in temperate grasses, to create corn and rice plants that are tolerant to a range of biotic and abiotic stresses, and for improved management of post-harvest control. We argue that practices used in plant breeding, seed treatments and agriculture, often caused by poor knowledge of the importance of fungal endophytes, are among the reasons for the loss of fungal endophyte diversity in domesticated plants and also accounts for the reduced effectiveness of some endophyte strains to confer plant benefits. We provide recommendations on how to mitigate against these negative impacts in modern agriculture. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Phylogenetic analysis of fungal ABC transporters

    Directory of Open Access Journals (Sweden)

    Driessen Arnold JM

    2010-03-01

    Full Text Available Abstract Background The superfamily of ABC proteins is among the largest known in nature. Its members are mainly, but not exclusively, involved in the transport of a broad range of substrates across biological membranes. Many contribute to multidrug resistance in microbial pathogens and cancer cells. The diversity of ABC proteins in fungi is comparable with those in multicellular animals, but so far fungal ABC proteins have barely been studied. Results We performed a phylogenetic analysis of the ABC proteins extracted from the genomes of 27 fungal species from 18 orders representing 5 fungal phyla thereby covering the most important groups. Our analysis demonstrated that some of the subfamilies of ABC proteins remained highly conserved in fungi, while others have undergone a remarkable group-specific diversification. Members of the various fungal phyla also differed significantly in the number of ABC proteins found in their genomes, which is especially reduced in the yeast S. cerevisiae and S. pombe. Conclusions Data obtained during our analysis should contribute to a better understanding of the diversity of the fungal ABC proteins and provide important clues about their possible biological functions.

  12. Selective Inhibition of Steroidogenic Enzymes by Ketoconazole in Rat Ovary Cells

    Directory of Open Access Journals (Sweden)

    Michael Gal

    2014-01-01

    Full Text Available Objective Ketoconazole (KCZ is an anti-fungal agent extensively used for clinical applications related to its inhibitory effects on adrenal and testicular steroidogenesis. Much less information is available on the effects of KCZ on synthesis of steroid hormones in the ovary. The present study aimed to characterize the in situ effects of KCZ on steroidogenic enzymes in primary rat ovary cells. Methods Following the induction of folliculogenesis in gonadotropin treated rats, freshly prepared ovarian cells were incubated in suspension for up to four hours while radiolabeled steroid substrates were added and time dependent generation of their metabolic products was analyzed by thin layer chromatography (TLC. Results KCZ inhibits the P450 steroidogenic enzymes in a selective and dose dependent manner, including cholesterol side-chain cleavage cytochrome P450 (CYP11A1/P450scc, the 17α-hydroxylase activity of CYP17A1/P450c17, and CYP19A1/P450arom, with IC 50 values of 0.3, 1.8, and 0.3 μg/mL (0.56, 3.36, and 0.56 μM, respectively. Unaffected by KCZ, at 10 μg/mL, were the 17,20 lyase activity of CYP17A1, as well as five non-cytochrome steroidogenic enzymes including 3β-hydroxysteroid dehydrogenase-δ 5-4 isomerase type 1 (3βHSD1, 5α-reductase, 20α-hydroxysteroid dehydrogenase (20α-HSD, 3α-hydroxysteroid dehydrogenase (3α-HSD, and 17β-hydroxysteroid dehydrogenase type 1 (17HSD1. Conclusion These findings map the effects of KCZ on the ovarian pathways of progestin, androgen, and estrogen synthesis. Hence, the drug may have a potential use as an acute and reversible modulator of ovarian steroidogenesis in pathological circumstances.

  13. Potential Roles of Fungal Extracellular Vesicles during Infection

    Science.gov (United States)

    Joffe, Luna S.; Nimrichter, Leonardo

    2016-01-01

    ABSTRACT Extracellular vesicles (EVs) are produced by virtually all cell types. Within the past few years, work in this field has revealed more information about fungal EVs. Fungal EVs have been shown to carry proteins, lipids, pigments, polysaccharides, and RNA; these components are known virulence factors, a fact which supports the hypothesis that fungal EVs concentrate pathogenic determinants. Additionally, recent studies have demonstrated that fungal EVs stimulate the host immune system. In this review, putative roles of fungal EVs are discussed, including their potential as vaccination tools and their possible contribution to pathogenesis in invasive fungal diseases. PMID:27390779

  14. Fungal demethylation of Kraft lignin.

    Science.gov (United States)

    Zou, Linyou; Ross, Brian M; Hutchison, Leonard J; Christopher, Lew P; Dekker, Robert F H; Malek, Lada

    2015-06-01

    Demethylation of industrial lignin has been for long coveted as a pathway to the production of an abundant natural substitute for fossil-oil derived phenol. In an attempt to possibly identify a novel Kraft lignin-demethylating enzyme, we surveyed a collection of fungi by using selected ion flow tube-mass spectrometry (SIFT-MS). This method readily identifies methanol resulting from lignin demethylation activity. Absidia cylindrospora, and unidentified Cylindrocladium sp. and Aspergillus sp. were shown to metabolize lignin via different pathways, based on the HPLC analysis of lignin fragments. Of these three, Cylindrocladium and Aspergillus were shown to retain most of the lignin intact after 3 weeks in culture, while removing about 40% of the available methoxy groups. Our results demonstrate that after optimization of culture and lignin recovery methods, biological modification of Kraft lignin may be a feasible pathway to obtaining demethylated lignin for further industrial use. Crown Copyright © 2015. Published by Elsevier Inc. All rights reserved.

  15. Chitinolytic Enterobacter agglomerans Antagonistic to Fungal Plant Pathogens.

    Science.gov (United States)

    Chernin, L; Ismailov, Z; Haran, S; Chet, I

    1995-05-01

    Three Enterobacter agglomerans strains which produce and excrete proteins with chitinolytic activity were found while screening soil-borne bacteria antagonistic to fungal plant pathogens. The chitinolytic activity was induced when the strains were grown in the presence of colloidal chitin as the sole carbon source. It was quantitated by using assays with chromogenic p-nitrophenyl analogs of disaccharide, trisaccharide, and tetrasaccharide derivatives of N-acetylglucosamine. A set of three fluorescent substrates with a 4-methylumbelliferyl group linked by (beta)-1,4 linkage to N-acetylglucosamine mono- or oligosaccharides were used to identify the chitinolytic activities of proteins which had been renatured following their separation by electrophoresis. This study provides the most complete evidence for the presence of a complex of chitinolytic enzymes in Enterobacter strains. Four enzymes were detected: two N-acetyl-(beta)-d-glucosaminidases of 89 and 67 kDa, an endochitinase with an apparent molecular mass of 59 kDa, and a chitobiosidase of 50 kDa. The biocontrol ability of the chitinolytic strains was demonstrated under greenhouse conditions. The bacteria decreased the incidence of disease caused by Rhizoctonia solani in cotton by 64 to 86%. Two Tn5 mutants of one of the isolates, which were deficient in chitinolytic activity, were unable to protect plants against the disease.

  16. Fungal Chitin Dampens Inflammation through IL-10 Induction Mediated by NOD2 and TLR9 Activation

    Science.gov (United States)

    Wagener, Jeanette; Malireddi, R. K. Subbarao; Lenardon, Megan D.; Köberle, Martin; Vautier, Simon; MacCallum, Donna M.; Biedermann, Tilo; Schaller, Martin; Netea, Mihai G.; Kanneganti, Thirumala-Devi; Brown, Gordon D.; Brown, Alistair J. P.; Gow, Neil A. R.

    2014-01-01

    during fungal infection and restore immune balance after an infection has been cleared. By increasing the chitin content in the cell wall pathogenic fungi may influence the immune system in their favour, by down-regulating protective inflammatory immune responses. Furthermore, gene mutations and dysregulated enzyme activity in the described chitin recognition pathway are implicated in inflammatory conditions such as Crohn's Disease and asthma, highlighting the importance of the discovered mechanism in human health. PMID:24722226

  17. Improvement in Saccharification Yield of Mixed Rumen Enzymes by Identification of Recalcitrant Cell Wall Constituents Using Enzyme Fingerprinting

    Directory of Open Access Journals (Sweden)

    Ajay Badhan

    2015-01-01

    Full Text Available Identification of recalcitrant factors that limit digestion of forages and the development of enzymatic approaches that improve hydrolysis could play a key role in improving the efficiency of meat and milk production in ruminants. Enzyme fingerprinting of barley silage fed to heifers and total tract indigestible fibre residue (TIFR collected from feces was used to identify cell wall components resistant to total tract digestion. Enzyme fingerprinting results identified acetyl xylan esterases as key to the enhanced ruminal digestion. FTIR analysis also suggested cross-link cell wall polymers as principal components of indigested fiber residues in feces. Based on structural information from enzymatic fingerprinting and FTIR, enzyme pretreatment to enhance glucose yield from barley straw and alfalfa hay upon exposure to mixed rumen-enzymes was developed. Prehydrolysis effects of recombinant fungal fibrolytic hydrolases were analyzed using microassay in combination with statistical experimental design. Recombinant hemicellulases and auxiliary enzymes initiated degradation of plant structural polysaccharides upon application and improved the in vitro saccharification of alfalfa and barley straw by mixed rumen enzymes. The validation results showed that microassay in combination with statistical experimental design can be successfully used to predict effective enzyme pretreatments that can enhance plant cell wall digestion by mixed rumen enzymes.

  18. Improvement in Saccharification Yield of Mixed Rumen Enzymes by Identification of Recalcitrant Cell Wall Constituents Using Enzyme Fingerprinting.

    Science.gov (United States)

    Badhan, Ajay; Wang, Yu-Xi; Gruninger, Robert; Patton, Donald; Powlowski, Justin; Tsang, Adrian; McAllister, Tim A

    2015-01-01

    Identification of recalcitrant factors that limit digestion of forages and the development of enzymatic approaches that improve hydrolysis could play a key role in improving the efficiency of meat and milk production in ruminants. Enzyme fingerprinting of barley silage fed to heifers and total tract indigestible fibre residue (TIFR) collected from feces was used to identify cell wall components resistant to total tract digestion. Enzyme fingerprinting results identified acetyl xylan esterases as key to the enhanced ruminal digestion. FTIR analysis also suggested cross-link cell wall polymers as principal components of indigested fiber residues in feces. Based on structural information from enzymatic fingerprinting and FTIR, enzyme pretreatment to enhance glucose yield from barley straw and alfalfa hay upon exposure to mixed rumen-enzymes was developed. Prehydrolysis effects of recombinant fungal fibrolytic hydrolases were analyzed using microassay in combination with statistical experimental design. Recombinant hemicellulases and auxiliary enzymes initiated degradation of plant structural polysaccharides upon application and improved the in vitro saccharification of alfalfa and barley straw by mixed rumen enzymes. The validation results showed that microassay in combination with statistical experimental design can be successfully used to predict effective enzyme pretreatments that can enhance plant cell wall digestion by mixed rumen enzymes.

  19. A novel GH10 xylanase fromPenicilliumsp. accelerates saccharification of alkaline-pretreated bagasse by an enzyme from recombinantTrichoderma reeseiexpressingAspergillusβ-glucosidase.

    Science.gov (United States)

    Shibata, Nozomu; Suetsugu, Mari; Kakeshita, Hiroshi; Igarashi, Kazuaki; Hagihara, Hiroshi; Takimura, Yasushi

    2017-01-01

    Trichoderma reesei is considered a candidate fungal enzyme producer for the economic saccharification of cellulosic biomass. However, performance of the saccharifying enzymes produced by T. reesei is insufficient. Therefore, many attempts have been made to improve its performance by heterologous protein expression. In this study, to increase the conversion efficiency of alkaline-pretreated bagasse to sugars, we conducted screening of biomass-degrading enzymes that showed synergistic effects with enzyme preparations produced by recombinant T. reesei . Penicillium sp. strain KSM-F532 produced the most effective enzyme to promote the saccharification of alkaline-pretreated bagasse. Biomass-degrading enzymes from strain KSM-F532 were fractionated and analyzed, and a xylanase, named PspXyn10, was identified. The amino acid sequence of PspXyn10 was determined by cDNA analysis: the enzyme shows a modular structure consisting of glycoside hydrolase family 10 (GH10) and carbohydrate-binding module family 1 (CBM1) domains. Purified PspXyn10 was prepared from the supernatant of a recombinant T. reesei strain. The molecular weight of PspXyn10 was estimated to be 55 kDa, and its optimal temperature and pH for xylanase activity were 75 °C and pH 4.5, respectively. More than 80% of the xylanase activity was maintained at 65 °C for 10 min. With beechwood xylan as the substrate, the enzyme had a K m of 2.2 mg/mL and a V max of 332 μmol/min/mg. PspXyn10ΔCBM, which lacked the CBM1 domain, was prepared by limited proteolysis. PspXyn10ΔCBM showed increased activity against soluble xylan, but decreased saccharification efficiency of alkaline-pretreated bagasse. This result indicated that the CBM1 domain of PspXyn10 contributes to the enhancement of the saccharification efficiency of alkaline-pretreated bagasse. A recombinant T. reesei strain, named X2PX10, was constructed from strain X3AB1. X3AB1 is an Aspergillus aculeatus β-glucosidase-expressing T. reesei PC-3-7. X2PX10

  20. Immunological Consequences of Intestinal Fungal Dysbiosis.

    Science.gov (United States)

    Wheeler, Matthew L; Limon, Jose J; Bar, Agnieszka S; Leal, Christian A; Gargus, Matthew; Tang, Jie; Brown, Jordan; Funari, Vincent A; Wang, Hanlin L; Crother, Timothy R; Arditi, Moshe; Underhill, David M; Iliev, Iliyan D

    2016-06-08

    Compared to bacteria, the role of fungi within the intestinal microbiota is poorly understood. In this study we investigated whether the presence of a "healthy" fungal community in the gut is important for modulating immune function. Prolonged oral treatment of mice with antifungal drugs resulted in increased disease severity in acute and chronic models of colitis, and also exacerbated the development of allergic airway disease. Microbiota profiling revealed restructuring of fungal and bacterial communities. Specifically, representation of Candida spp. was reduced, while Aspergillus, Wallemia, and Epicoccum spp. were increased. Oral supplementation with a mixture of three fungi found to expand during antifungal treatment (Aspergillus amstelodami, Epicoccum nigrum, and Wallemia sebi) was sufficient to recapitulate the exacerbating effects of antifungal drugs on allergic airway disease. Taken together, these results indicate that disruption of commensal fungal populations can influence local and peripheral immune responses and enhance relevant disease states. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Direct Surface Analysis of Fungal Species by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Valentine, Nancy B.(BATTELLE (PACIFIC NW LAB)); Wahl, Jon H.(BATTELLE (PACIFIC NW LAB)); Kingsley, Mark T.(BATTELLE (PACIFIC NW LAB)); Wahl, Karen L.(BATTELLE (PACIFIC NW LAB))

    2001-12-01

    Intact spores and/or hyphae of Aspergillus niger, Rhizopus oryzae, Trichoderma reesei and Phanerochaete chrysosporium are analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). This study investigates various methods of sample preparation and matrices to determine optimum collection and analysis criteria for fungal analysis by MALDI-MS. Fungi are applied to the MALDI sample target as untreated, sonicated, acid/heat treated, or blotted directly from the fungal culture with double-stick tape. Ferulic acid or sinapinic acid matrix solution is layered over the dried samples and analyzed by MALDI-MS. Statistical analysis of the data show that simply using double stick tape to collect and transfer to a MALDI sample plate typically worked as well as the other preparation methods, but requires the least sample handling.

  2. Enzymes for improved biomass conversion

    Science.gov (United States)

    Brunecky, Roman; Himmel, Michael E.

    2016-02-02

    Disclosed herein are enzymes and combinations of the enzymes useful for the hydrolysis of cellulose and the conversion of biomass. Methods of degrading cellulose and biomass using enzymes and cocktails of enzymes are also disclosed.

  3. Identification & Characterization of Fungal Ice Nucleation Proteins

    Science.gov (United States)

    Scheel, Jan Frederik; Kunert, Anna Theresa; Kampf, Christopher Johannes; Mauri, Sergio; Weidner, Tobias; Pöschl, Ulrich; Fröhlich-Nowoisky, Janine

    2016-04-01

    Freezing of water at relatively warm subfreezing temperatures is dependent on ice nucleation catalysis facilitated by ice nuclei (IN). These IN can be of various origins and although extensive research was done and progress was achieved, the nature and mechanisms leading to an effective IN are to date still poorly understood. Some of the most important processes of our geosphere like the water cycle are highly dependent on effective ice nucleation at temperatures between -2°C - -8°C, a temperature range which is almost exclusively covered by biological IN (BioIN). BioIN are usually macromolecular structures of biological polymers. Sugars as well as proteins have been reported to serve as IN and the best characterized BioIN are ice nucleation proteins (IN-P) from gram negative bacteria. Fungal strains from Fusarium spp. were described to be effective IN at subfreezing temperatures up to -2°C already 25 years ago and more and more fungal species are described to serve as efficient IN. Fungal IN are also thought to be proteins or at least contain a proteinaceous compound, but to date the fungal IN-P primary structure as well as their coding genetic elements of all IN active fungi are unknown. The aim of this study is a.) to identify the proteins and their coding genetic elements from IN active fungi (F. acuminatum, F. avenaceum, M. alpina) and b.) to characterize the mechanisms by which fungal IN serve as effective IN. We designed an interdisciplinary approach using biological, analytical and physical methods to identify fungal IN-P and describe their biological, chemical, and physical properties.

  4. Substrate analogues for isoprenoid enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Stremler, K.E.

    1987-01-01

    Diphosphonate analogues of geranyl diphosphate, resistant to degradation by phosphatases, were found to be alternate substrates for the reaction with farnesyl diphosphate synthetase isolated from avian liver. The difluoromethane analogue was shown to be the better alternate substrate, in agreement with solvolysis results which indicate that the electronegativity of the difluoromethylene unit more closely approximates that of the normal bridging oxygen. The usefulness of the C/sub 10/ difluoro analogue, for detecting low levels of isoprenoid enzymes in the presence of high levels of phosphatase activity, was demonstrated with a cell-free preparation from lemon peel. A series of C/sub 5/ through C/sub 15/ homoallylic and allylic diphosphonates, as well as two 5'-nucleotide diphosphonates, was prepared in high overall yield using the activation-displacement sequence. Radiolabeled samples of several of the allylic diphosphonates were prepared with tritium located at C1. A series of geraniols, stereospecifically deuterated at C1, was prepared. The enantiomeric purities and absolute configurations were determined by derivatization as the mandelate esters for analysis by /sup 1/H NMR. The stereochemistry of the activation-displacement sequence was examined using C1-deuterated substrates.

  5. Production of lignocellulolytic enzymes from three white-rot fungi by ...

    African Journals Online (AJOL)

    The objective of the work was to evaluate the effect of the media formulation on the production of lignocellulolytic enzymes and degradation of lignocellulosic components by the three fungal species. C. versicolor exhibited the highest ability to degrade the three main polymers of the lignocellulosic waste materials employed ...

  6. Enzymes in animal nutrition

    OpenAIRE

    Scientific Committee on Animal Nutrition

    2011-01-01

    This report brings overview of endogenous as well as exogenous enzymes and their role and importance in animal nutrition. Enzymes for animal nutrition have been systematically developed since 1980´s. Phytase, xylanase and β-glucanase are used in poultry-rising, pig breeding, aquaculture and begin to push to the ruminant nutrition. Phytase increase availability of P, Ca, Zn, digestibility of proteins and fats. Its positive effect on the environment is well described – enzymes decrease the cont...

  7. Profiling the orphan enzymes

    Science.gov (United States)

    2014-01-01

    The emergence of Next Generation Sequencing generates an incredible amount of sequence and great potential for new enzyme discovery. Despite this huge amount of data and the profusion of bioinformatic methods for function prediction, a large part of known enzyme activities is still lacking an associated protein sequence. These particular activities are called “orphan enzymes”. The present review proposes an update of previous surveys on orphan enzymes by mining the current content of public databases. While the percentage of orphan enzyme activities has decreased from 38% to 22% in ten years, there are still more than 1,000 orphans among the 5,000 entries of the Enzyme Commission (EC) classification. Taking into account all the reactions present in metabolic databases, this proportion dramatically increases to reach nearly 50% of orphans and many of them are not associated to a known pathway. We extended our survey to “local orphan enzymes” that are activities which have no representative sequence in a given clade, but have at least one in organisms belonging to other clades. We observe an important bias in Archaea and find that in general more than 30% of the EC activities have incomplete sequence information in at least one superkingdom. To estimate if candidate proteins for local orphans could be retrieved by homology search, we applied a simple strategy based on the PRIAM software and noticed that candidates may be proposed for an important fraction of local orphan enzymes. Finally, by studying relation between protein domains and catalyzed activities, it appears that newly discovered enzymes are mostly associated with already known enzyme domains. Thus, the exploration of the promiscuity and the multifunctional aspect of known enzyme families may solve part of the orphan enzyme issue. We conclude this review with a presentation of recent initiatives in finding proteins for orphan enzymes and in extending the enzyme world by the discovery of new

  8. The EBI enzyme portal

    OpenAIRE

    Alc?ntara, Rafael; Onwubiko, Joseph; Cao, Hong; de Matos, Paula; Cham, Jennifer A.; Jacobsen, Jules; Holliday, Gemma L.; Fischer, Julia D.; Rahman, Syed Asad; Jassal, Bijay; Goujon, Mikael; Rowland, Francis; Velankar, Sameer; L?pez, Rodrigo; Overington, John P.

    2012-01-01

    The availability of comprehensive information about enzymes plays an important role in answering questions relevant to interdisciplinary fields such as biochemistry, enzymology, biofuels, bioengineering and drug discovery. At the EMBL European Bioinformatics Institute, we have developed an enzyme portal (http://www.ebi.ac.uk/enzymeportal) to provide this wealth of information on enzymes from multiple in-house resources addressing particular data classes: protein sequence and structure, reacti...

  9. Enzyme catalysed tandem reactions

    OpenAIRE

    Oroz-Guinea, Isabel; García-Junceda, Eduardo

    2013-01-01

    To transfer to the laboratory, the excellent efficiency shown by enzymes in Nature, biocatalysis, had to mimic several synthetic strategies used by the living organisms. Biosynthetic pathways are examples of tandem catalysis and may be assimilated in the biocatalysis field for the use of isolated multi-enzyme systems in the homogeneous phase. The concurrent action of several enzymes that work sequentially presents extraordinary advantages from the synthetic point of view, since it permits a r...

  10. Fungal nail infections: diagnosis and management.

    Science.gov (United States)

    2009-02-01

    1) Fungal nail infection, or onychomycosis, mainly affects toenails. Infections are generally asymptomatic. Spontaneous regressions, but also complications, appear to be rare. Discomfort and cosmetic complaint are occasionally reported; 2) After a review of the literature based on the standard Prescrire procedure, we examined the diagnosis and management of fungal nail infections; 3) Clinical signs of fungal nail infections are non-specific. Alternative diagnoses include psoriasis and nail microtrauma. Nail hyperkeratosis and leukonychia are useful diagnostic pointers. Matrix involvement has important implications in the choice of treatment; 4) Detection of fungal structures by direct examination of a nail sample is strongly suggestive of fungal nail infection. In contrast, cases of negative direct examination with positive culture must be interpreted with caution, as contamination is frequent; 5) Antifungal lacquers (5% amorolfine and 8% ciclopirox) applied to the nails cure about 30% of fungal infections and sometimes cause mild irritation. There is no firm evidence that these solutions are any more effective than other topical antifungals applied daily to the affected nail. Trimming, filing or grinding the nail, in addition to these drug treatments, is likely to be beneficial, but these measures have not been evaluated; 6) Chemical nail destruction with a combination of urea and bifonazole, followed by treatment with an antifungal ointment, can be used when the nail is markedly thickened. Non-comparative trials have shown cure rates close to 70% at three months when the matrix is not involved, and 40% with matrix involvement. Drug application is inconvenient and local reactions are frequent. Surgical nail avulsion carries a risk of local infection and permanent nail dystrophy; 7) Oral terbinafine is effective in more than 50% of cases but its cutaneous, hepatic and haematological adverse effects are severe in about 1 in 2000 patients and can be life

  11. Fungal infections of the lung in children

    Energy Technology Data Exchange (ETDEWEB)

    Toma, Paolo; Colafati, Giovanna Stefania; D' Andrea, Maria Luisa [IRCCS Bambino Gesu Children' s Hospital, Department of Imaging, Rome (Italy); Bertaina, Alice; Mastronuzzi, Angela [IRCCS Bambino Gesu Children' s Hospital, Department of Pediatric Hematology/Oncology and Transfusion Medicine, Rome (Italy); Castagnola, Elio [IRCCS Istituto Giannina Gaslini, Department of Infective Diseases, Genoa (Italy); Finocchi, Andrea [IRCCS Bambino Gesu Children' s Hospital, Department of Pediatrics, Rome (Italy); Lucidi, Vincenzina [IRCCS Bambino Gesu Children' s Hospital, Cystic Fibrosis Center, Rome (Italy); Granata, Claudio [IRCCS Istituto Giannina Gaslini, Department of Pediatric Radiology, Genoa (Italy)

    2016-12-15

    Fungal infections of the lungs are relatively common and potentially life-threatening conditions in immunocompromised children. The role of imaging in children with lung mycosis is to delineate the extension of pulmonary involvement, to assess response to therapy, and to monitor for adverse sequelae such as bronchiectasis and cavitation. The aim of this paper is to show imaging findings in a series of patients with fungal pneumonia from two tertiary children's hospitals, to discuss differential diagnoses and to show how imaging findings can vary depending on the host immune response. (orig.)

  12. Enzymes from extremophiles.

    Science.gov (United States)

    Demirjian, D C; Morís-Varas, F; Cassidy, C S

    2001-04-01

    The industrial application of enzymes that can withstand harsh conditions has greatly increased over the past decade. This is mainly a result of the discovery of novel enzymes from extremophilic microorganisms. Recent advances in the study of extremozymes point to the acceleration of this trend. In particular, enzymes from thermophilic organisms have found the most practical commercial use to date because of their overall inherent stability. This has also led to a greater understanding of stability factors involved in adaptation of these enzymes to their unusual environments.

  13. Artificial Enzymes, "Chemzymes"

    DEFF Research Database (Denmark)

    Bjerre, Jeannette; Rousseau, Cyril Andre Raphaël; Pedersen, Lavinia Georgeta M

    2008-01-01

    "Chemzymes", based on cyclodextrins and other molecules. Only the chemzymes that have shown enzyme-like activity that has been quantified by different methods will be mentioned. This review will summarize the work done in the field of artificial glycosidases, oxidases, epoxidases, and esterases, as well......Enzymes have fascinated scientists since their discovery and, over some decades, one aim in organic chemistry has been the creation of molecules that mimic the active sites of enzymes and promote catalysis. Nevertheless, even today, there are relatively few examples of enzyme models...

  14. The purification and characterization of ATP synthase complexes from the mitochondria of four fungal species.

    Science.gov (United States)

    Liu, Sidong; Charlesworth, Thomas J; Bason, John V; Montgomery, Martin G; Harbour, Michael E; Fearnley, Ian M; Walker, John E

    2015-05-15

    The ATP synthases have been isolated by affinity chromatography from the mitochondria of the fungal species Yarrowia lipolytica, Pichia pastoris, Pichia angusta and Saccharomyces cerevisiae. The subunit compositions of the purified enzyme complexes depended on the detergent used to solubilize and purify the complex, and the presence or absence of exogenous phospholipids. All four enzymes purified in the presence of n-dodecyl-β-D-maltoside had a complete complement of core subunits involved directly in the synthesis of ATP, but they were deficient to different extents in their supernumerary membrane subunits. In contrast, the enzymes from P. angusta and S. cerevisiae purified in the presence of n-decyl-β-maltose neopentyl glycol and the phospholipids 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine, cardiolipin (diphosphatidylglycerol) and 1-palmitoyl-2-oleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] had a complete complement of core subunits and also contained all of the known supernumerary membrane subunits, e, f, g, j, k and ATP8 (or Aap1), plus an additional new membrane component named subunit l, related in sequence to subunit k. The catalytic domain of the enzyme from P. angusta was more resistant to thermal denaturation than the enzyme from S. cerevisiae, but less stable than the catalytic domain of the bovine enzyme, but the stator and the integrity of the transmembrane proton pathway were most stable in the enzyme from P. angusta. The P. angusta enzyme provides a suitable source of enzyme for studying the structure of the membrane domain and properties associated with that sector of the enzyme complex.

  15. Structural Diversity and Biological Activities of Fungal Cyclic Peptides, Excluding Cyclodipeptides.

    Science.gov (United States)

    Wang, Xiaohan; Lin, Minyi; Xu, Dan; Lai, Daowan; Zhou, Ligang

    2017-11-27

    Cyclic peptides are cyclic compounds formed mainly by the amide bonds between either proteinogenic or non-proteinogenic amino acids. This review highlights the occurrence, structures and biological activities of fungal cyclic peptides (excluding cyclodipeptides, and peptides containing ester bonds in the core ring) reported until August 2017. About 293 cyclic peptides belonging to the groups of cyclic tri-, tetra-, penta-, hexa-, hepta-, octa-, nona-, deca-, undeca-, dodeca-, tetradeca-, and octadecapeptides as well as cyclic peptides containing ether bonds in the core ring have been isolated from fungi. They were mainly isolated from the genera Aspergillus , Penicillium , Fusarium , Acremonium and Amanita . Some of them were screened to have antimicrobial, antiviral, cytotoxic, phytotoxic, insecticidal, nematicidal, immunosuppressive and enzyme-inhibitory activities to show their potential applications. Some fungal cyclic peptides such as the echinocandins, pneumocandins and cyclosporin A have been developed as pharmaceuticals.

  16. Structural Diversity and Biological Activities of Fungal Cyclic Peptides, Excluding Cyclodipeptides

    Directory of Open Access Journals (Sweden)

    Xiaohan Wang

    2017-11-01

    Full Text Available Cyclic peptides are cyclic compounds formed mainly by the amide bonds between either proteinogenic or non-proteinogenic amino acids. This review highlights the occurrence, structures and biological activities of fungal cyclic peptides (excluding cyclodipeptides, and peptides containing ester bonds in the core ring reported until August 2017. About 293 cyclic peptides belonging to the groups of cyclic tri-, tetra-, penta-, hexa-, hepta-, octa-, nona-, deca-, undeca-, dodeca-, tetradeca-, and octadecapeptides as well as cyclic peptides containing ether bonds in the core ring have been isolated from fungi. They were mainly isolated from the genera Aspergillus, Penicillium, Fusarium, Acremonium and Amanita. Some of them were screened to have antimicrobial, antiviral, cytotoxic, phytotoxic, insecticidal, nematicidal, immunosuppressive and enzyme-inhibitory activities to show their potential applications. Some fungal cyclic peptides such as the echinocandins, pneumocandins and cyclosporin A have been developed as pharmaceuticals.

  17. Molecular docking and dynamics simulation analyses unraveling the differential enzymatic catalysis by plant and fungal laccases with respect to lignin biosynthesis and degradation.

    Science.gov (United States)

    Awasthi, Manika; Jaiswal, Nivedita; Singh, Swati; Pandey, Veda P; Dwivedi, Upendra N

    2015-09-01

    Laccase, widely distributed in bacteria, fungi, and plants, catalyzes the oxidation of wide range of compounds. With regards to one of the important physiological functions, plant laccases are considered to catalyze lignin biosynthesis while fungal laccases are considered for lignin degradation. The present study was undertaken to explain this dual function of laccases using in-silico molecular docking and dynamics simulation approaches. Modeling and superimposition analyses of one each representative of plant and fungal laccases, namely, Populus trichocarpa and Trametes versicolor, respectively, revealed low level of similarity in the folding of two laccases at 3D levels. Docking analyses revealed significantly higher binding efficiency for lignin model compounds, in proportion to their size, for fungal laccase as compared to that of plant laccase. Residues interacting with the model compounds at the respective enzyme active sites were found to be in conformity with their role in lignin biosynthesis and degradation. Molecular dynamics simulation analyses for the stability of docked complexes of plant and fungal laccases with lignin model compounds revealed that tetrameric lignin model compound remains attached to the active site of fungal laccase throughout the simulation period, while it protrudes outwards from the active site of plant laccase. Stability of these complexes was further analyzed on the basis of binding energy which revealed significantly higher stability of fungal laccase with tetrameric compound than that of plant. The overall data suggested a situation favorable for the degradation of lignin polymer by fungal laccase while its synthesis by plant laccase.

  18. Indoor airborne fungal pollution in newborn units in Turkey

    NARCIS (Netherlands)

    Demirel, Rasime; Sen, Burhan; Kadaifciler, Duygu; Yoltas, Aysegul; Okten, Suzan; Ozkale, Evrim; Berikten, Derya; Samson, Robert A.; Haliki Uztan, Alev; Yilmaz, Neriman; Abaci Gunyar, Ozlem; Aydogdu, Halide; Asan, Ahmet; Kivanc, Merih; Ozdil, Soner; Sakartepe, Erhan

    2017-01-01

    Pathogenic and/or opportunistic fungal species are major causes of nosocomial infections, especially in controlled environments where immunocompromised patients are hospitalized. Indoor fungal contamination in hospital air is associated with a wide range of adverse health effects. Regular

  19. Fungal isolates and their toxicity from different ecosystems in ...

    African Journals Online (AJOL)

    ONOS

    2010-08-23

    , chemical and drug industries. Since then, new fungal ...... Nesci A, Barros G, Castillo C, Etcheverry M (2006). Soil fungal population in preharvest maize ecosystem in different tillage practices in Argentina. Soil Tillage Res.

  20. MycoCosm, an Integrated Fungal Genomics Resource

    Energy Technology Data Exchange (ETDEWEB)

    Shabalov, Igor; Grigoriev, Igor

    2012-03-16

    MycoCosm is a web-based interactive fungal genomics resource, which was first released in March 2010, in response to an urgent call from the fungal community for integration of all fungal genomes and analytical tools in one place (Pan-fungal data resources meeting, Feb 21-22, 2010, Alexandria, VA). MycoCosm integrates genomics data and analysis tools to navigate through over 100 fungal genomes sequenced at JGI and elsewhere. This resource allows users to explore fungal genomes in the context of both genome-centric analysis and comparative genomics, and promotes user community participation in data submission, annotation and analysis. MycoCosm has over 4500 unique visitors/month or 35000+ visitors/year as well as hundreds of registered users contributing their data and expertise to this resource. Its scalable architecture allows significant expansion of the data expected from JGI Fungal Genomics Program, its users, and integration with external resources used by fungal community.

  1. Comparative genomic analysis of human fungal pathogens causing paracoccidioidomycosis.

    Directory of Open Access Journals (Sweden)

    Christopher A Desjardins

    2011-10-01

    Full Text Available Paracoccidioides is a fungal pathogen and the cause of paracoccidioidomycosis, a health-threatening human systemic mycosis endemic to Latin America. Infection by Paracoccidioides, a dimorphic fungus in the order Onygenales, is coupled with a thermally regulated transition from a soil-dwelling filamentous form to a yeast-like pathogenic form. To better understand the genetic basis of growth and pathogenicity in Paracoccidioides, we sequenced the genomes of two strains of Paracoccidioides brasiliensis (Pb03 and Pb18 and one strain of Paracoccidioides lutzii (Pb01. These genomes range in size from 29.1 Mb to 32.9 Mb and encode 7,610 to 8,130 genes. To enable genetic studies, we mapped 94% of the P. brasiliensis Pb18 assembly onto five chromosomes. We characterized gene family content across Onygenales and related fungi, and within Paracoccidioides we found expansions of the fungal-specific kinase family FunK1. Additionally, the Onygenales have lost many genes involved in carbohydrate metabolism and fewer genes involved in protein metabolism, resulting in a higher ratio of proteases to carbohydrate active enzymes in the Onygenales than their relatives. To determine if gene content correlated with growth on different substrates, we screened the non-pathogenic onygenale Uncinocarpus reesii, which has orthologs for 91% of Paracoccidioides metabolic genes, for growth on 190 carbon sources. U. reesii showed growth on a limited range of carbohydrates, primarily basic plant sugars and cell wall components; this suggests that Onygenales, including dimorphic fungi, can degrade cellulosic plant material in the soil. In addition, U. reesii grew on gelatin and a wide range of dipeptides and amino acids, indicating a preference for proteinaceous growth substrates over carbohydrates, which may enable these fungi to also degrade animal biomass. These capabilities for degrading plant and animal substrates suggest a duality in lifestyle that could enable pathogenic

  2. Potential Roles of Fungal Extracellular Vesicles during Infection

    OpenAIRE

    Joffe, Luna S.; Nimrichter, Leonardo; Rodrigues, Marcio L.; Del Poeta, Maurizio

    2016-01-01

    ABSTRACT Extracellular vesicles (EVs) are produced by virtually all cell types. Within the past few years, work in this field has revealed more information about fungal EVs. Fungal EVs have been shown to carry proteins, lipids, pigments, polysaccharides, and RNA; these components are known virulence factors, a fact which supports the hypothesis that fungal EVs concentrate pathogenic determinants. Additionally, recent studies have demonstrated that fungal EVs stimulate the host immune system. ...

  3. Drivers of soil fungal communities in boreal forests

    OpenAIRE

    Sterkenburg, Erica

    2016-01-01

    Boreal forests harbour diverse fungal communities with decisive roles in decomposition and plant nutrition. Difficulties in studying soil fungi have limited knowledge about how fungal communities are shaped. The objective of this thesis was to study factors influencing soil fungal communities, aiming for increased understanding of their effect on environmental processes. Using next generation sequencing, responses of fungal communities to their physical-chemical environment, and responses...

  4. Fungal community assemblage of different soil compartments in mangrove ecosystem

    OpenAIRE

    Loganathachetti, Dinesh Sanka; Poosakkannu, Anbu; Muthuraman, Sundararaman

    2017-01-01

    The fungal communities of different soil compartments in mangrove ecosystem are poorly studied. We sequenced the internal transcribed spacer (ITS) regions to characterize the fungal communities in Avicennia marina root-associated soils (rhizosphere and pneumatophore) and bulk soil compartments. The rhizosphere but not pneumatophore soil compartment had significantly lower fungal species richness than bulk soil. However, bulk soil fungal diversity (Shannon diversity index) was significantly hi...

  5. Immobilized enzyme reactor chromatography: Optimization of protein retention and enzyme activity in monolithic silica stationary phases

    International Nuclear Information System (INIS)

    Besanger, Travis R.; Hodgson, Richard J.; Green, James R.A.; Brennan, John D.

    2006-01-01

    Our group recently reported on the application of protein-doped monolithic silica columns for immobilized enzyme reactor chromatography, which allowed screening of enzyme inhibitors present in mixtures using mass spectrometry for detection. The enzyme was immobilized by entrapment within a bimodal meso/macroporous silica material prepared by a biocompatible sol-gel processing route. While such columns proved to be useful for applications such as screening of protein-ligand interactions, significant amounts of entrapped proteins leached from the columns owing to the high proportion of macropores within the materials. Herein, we describe a detailed study of factors affecting the morphology of protein-doped bioaffinity columns and demonstrate that specific pH values and concentrations of poly(ethylene glycol) can be used to prepare essentially mesoporous columns that retain over 80% of initially loaded enzyme in an active and accessible form and yet still retain sufficient porosity to allow pressure-driven flow in the low μL/min range. Using the enzyme γ-glutamyl transpeptidase (γ-GT), we further evaluated the catalytic constants of the enzyme entrapped in capillary columns with different silica morphologies as a function of flowrate and backpressure using the enzyme reactor assay mode. It was found that the apparent activity of the enzyme was highest in mesoporous columns that retained high levels of enzyme. In such columns, enzyme activity increased by ∼2-fold with increases in both flowrate (from 250 to 1000 nL/min) and backpressure generated (from 500 to 2100 psi) during the chromatographic activity assay owing to increases in k cat and decreases in K M , switching from diffusion controlled to reaction controlled conditions at ca. 2000 psi. These results suggest that columns with minimal macropore volumes (<5%) are advantageous for the entrapment of soluble proteins for bioaffinity and bioreactor chromatography

  6. Root-associated fungal microbiota of nonmycorrhizal Arabis alpina and its contribution to plant phosphorus nutrition.

    Science.gov (United States)

    Almario, Juliana; Jeena, Ganga; Wunder, Jörg; Langen, Gregor; Zuccaro, Alga; Coupland, George; Bucher, Marcel

    2017-10-31

    Most land plants live in association with arbuscular mycorrhizal (AM) fungi and rely on this symbiosis to scavenge phosphorus (P) from soil. The ability to establish this partnership has been lost in some plant lineages like the Brassicaceae, which raises the question of what alternative nutrition strategies such plants have to grow in P-impoverished soils. To understand the contribution of plant-microbiota interactions, we studied the root-associated fungal microbiome of Arabis alpina (Brassicaceae) with the hypothesis that some of its components can promote plant P acquisition. Using amplicon sequencing of the fungal internal transcribed spacer 2, we studied the root and rhizosphere fungal communities of A. alpina growing under natural and controlled conditions including low-P soils and identified a set of 15 fungal taxa consistently detected in its roots. This cohort included a Helotiales taxon exhibiting high abundance in roots of wild A. alpina growing in an extremely P-limited soil. Consequently, we isolated and subsequently reintroduced a specimen from this taxon into its native P-poor soil in which it improved plant growth and P uptake. The fungus exhibited mycorrhiza-like traits including colonization of the root endosphere and P transfer to the plant. Genome analysis revealed a link between its endophytic lifestyle and the expansion of its repertoire of carbohydrate-active enzymes. We report the discovery of a plant-fungus interaction facilitating the growth of a nonmycorrhizal plant under native P-limited conditions, thus uncovering a previously underestimated role of root fungal microbiota in P cycling.

  7. Indoleamine 2,3-dioxygenase controls fungal loads and immunity in Paracoccidioidomicosis but is more important to susceptible than resistant hosts.

    Science.gov (United States)

    Araújo, Eliseu F; Loures, Flávio V; Bazan, Silvia B; Feriotti, Claudia; Pina, Adriana; Schanoski, Alessandra S; Costa, Tânia A; Calich, Vera L G

    2014-11-01

    Paracoccidioidomycosis, a primary fungal infection restricted to Latin America, is acquired by inhalation of fungal particles. The immunoregulatory mechanisms that control the severe and mild forms of paracoccidioidomycosis are still unclear. Indoleamine 2,3-dioxygenase (IDO), an IFN-γ induced enzyme that catalyzes tryptophan metabolism, can control host-pathogen interaction by inhibiting pathogen growth, T cell immunity and tissue inflammation. In this study, we investigated the role of IDO in pulmonary paracoccidioidomycosis of susceptible and resistant mice. IDO was blocked by 1-methyl-dl-tryptophan (1MT), and fungal infection studied in vitro and in vivo. Paracoccidioides brasiliensis infection was more severe in 1MT treated than untreated macrophages of resistant and susceptible mice, concurrently with decreased production of kynurenines and IDO mRNA. Similar results were observed in the pulmonary infection. Independent of the host genetic pattern, IDO inhibition reduced fungal clearance but enhanced T cell immunity. The early IDO inhibition resulted in increased differentiation of dendritic and Th17 cells, accompanied by reduced responses of Th1 and Treg cells. Despite these equivalent biological effects, only in susceptible mice the temporary IDO blockade caused sustained fungal growth, increased tissue pathology and mortality rates. In contrast, resistant mice were able to recover the transitory IDO blockade by the late control of fungal burdens without enhanced tissue pathology. Our studies demonstrate for the first time that in pulmonary paracoccidioidomycosis, IDO is an important immunoregulatory enzyme that promotes fungal clearance and inhibits T cell immunity and inflammation, with prominent importance to susceptible hosts. In fact, only in the susceptible background IDO inhibition resulted in uncontrolled tissue pathology and mortality rates. Our findings open new perspectives to understand the immunopathology of paracoccidioidomycosis, and suggest

  8. Indoleamine 2,3-dioxygenase controls fungal loads and immunity in Paracoccidioidomicosis but is more important to susceptible than resistant hosts.

    Directory of Open Access Journals (Sweden)

    Eliseu F Araújo

    2014-11-01

    Full Text Available Paracoccidioidomycosis, a primary fungal infection restricted to Latin America, is acquired by inhalation of fungal particles. The immunoregulatory mechanisms that control the severe and mild forms of paracoccidioidomycosis are still unclear. Indoleamine 2,3-dioxygenase (IDO, an IFN-γ induced enzyme that catalyzes tryptophan metabolism, can control host-pathogen interaction by inhibiting pathogen growth, T cell immunity and tissue inflammation.In this study, we investigated the role of IDO in pulmonary paracoccidioidomycosis of susceptible and resistant mice. IDO was blocked by 1-methyl-dl-tryptophan (1MT, and fungal infection studied in vitro and in vivo. Paracoccidioides brasiliensis infection was more severe in 1MT treated than untreated macrophages of resistant and susceptible mice, concurrently with decreased production of kynurenines and IDO mRNA. Similar results were observed in the pulmonary infection. Independent of the host genetic pattern, IDO inhibition reduced fungal clearance but enhanced T cell immunity. The early IDO inhibition resulted in increased differentiation of dendritic and Th17 cells, accompanied by reduced responses of Th1 and Treg cells. Despite these equivalent biological effects, only in susceptible mice the temporary IDO blockade caused sustained fungal growth, increased tissue pathology and mortality rates. In contrast, resistant mice were able to recover the transitory IDO blockade by the late control of fungal burdens without enhanced tissue pathology.Our studies demonstrate for the first time that in pulmonary paracoccidioidomycosis, IDO is an important immunoregulatory enzyme that promotes fungal clearance and inhibits T cell immunity and inflammation, with prominent importance to susceptible hosts. In fact, only in the susceptible background IDO inhibition resulted in uncontrolled tissue pathology and mortality rates. Our findings open new perspectives to understand the immunopathology of

  9. Enzyme Vs. Extremozyme -32 ...

    Indian Academy of Sciences (India)

    Enzyme Vs. Extremozyme. What Makes Extremozymes Function Under Harsh Conditions? Santosh Kumar is ... extremozymes to high temperature or pH so that enzymes from mesophiles can be engineered to behave .... alkalinity (above pH 10, soda lake) from which extremozymes have been isolated. F C Lowyer of the ...

  10. Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi.

    Science.gov (United States)

    Gacura, Matthew D; Sprockett, Daniel D; Heidenreich, Bess; Blackwood, Christopher B

    2016-04-01

    Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. The primers were used to successfully amplify GH28 pectinases from all Ascomycota cultures tested, but only three out of seven Basidiomycota cultures. In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senesced tree leaves from different forest ecosystems, followed by cloning and sequencing. Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. All sequences obtained from GH28 primers were classified as Ascomycota; in contrast, ITS sequences indicated that fungal communities were up to 39% Basidiomycetes. Analysis of leaf samples indicated that both forest stand and ecosystem type were important in structuring fungal communities. However, site played the prominent role in explaining GH28 composition, whereas ecosystem type was more important for ITS composition, indicating possible genetic drift between populations of fungi. Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. High prevalence of a fungal prion

    NARCIS (Netherlands)

    Debets, A.J.M.; Dalstra, H.J.P.; Slakhorst, S.M.; Koopmanschap-Memelink, A.B.; Hoekstra, R.F.; Saupe, S.J.

    2012-01-01

    Prions are infectious proteins that cause fatal diseases in mammals. Prions have also been found in fungi, but studies on their role in nature are scarce. The proposed biological function of fungal prions is debated and varies from detrimental to benign or even beneficial. [Het-s] is a prion of the

  12. October 2012 Multistate Fungal Meningitis Outbreak

    Centers for Disease Control (CDC) Podcasts

    2012-10-17

    This podcast gives an overview of the October 2012 multistate fungal meningitis outbreak, including symptoms to watch for and a website for up-to-date information.  Created: 10/17/2012 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 10/17/2012.

  13. 50-plus years of fungal viruses

    Energy Technology Data Exchange (ETDEWEB)

    Ghabrial, Said A., E-mail: saghab00@email.uky.edu [Plant Pathology Department, University of Kentucky, Lexington, KY (United States); Castón, José R. [Department of Structure of Macromolecules, Centro Nacional Biotecnologıa/CSIC, Campus de Cantoblanco, Madrid (Spain); Jiang, Daohong [State Key Lab of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei Province (China); Nibert, Max L. [Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA (United States); Suzuki, Nobuhiro [Institute of Plant Science and Resources, Okayama University, Kurashiki, Okayama (Japan)

    2015-05-15

    Mycoviruses are widespread in all major taxa of fungi. They are transmitted intracellularly during cell division, sporogenesis, and/or cell-to-cell fusion (hyphal anastomosis), and thus their life cycles generally lack an extracellular phase. Their natural host ranges are limited to individuals within the same or closely related vegetative compatibility groups, although recent advances have established expanded experimental host ranges for some mycoviruses. Most known mycoviruses have dsRNA genomes packaged in isometric particles, but an increasing number of positive- or negative-strand ssRNA and ssDNA viruses have been isolated and characterized. Although many mycoviruses do not have marked effects on their hosts, those that reduce the virulence of their phytopathogenic fungal hosts are of considerable interest for development of novel biocontrol strategies. Mycoviruses that infect endophytic fungi and those that encode killer toxins are also of special interest. Structural analyses of mycoviruses have promoted better understanding of virus assembly, function, and evolution. - Highlights: • Historical perspective of fungal virus research. • Description, classification and diversity of fungal virus families. • Structural features of fungal virus particles. • Hypovirulence and exploitation of mycoviruses in biological control of plant pathogenic fungi.

  14. Thigmo Responses: The Fungal Sense of Touch.

    Science.gov (United States)

    Almeida, Mariana Cruz; Brand, Alexandra C

    2017-04-01

    The growth and development of most fungi take place on a two-dimensional surface or within a three-dimensional matrix. The fungal sense of touch is therefore critical for fungi in the interpretation of their environment and often signals the switch to a new developmental state. Contact sensing, or thigmo-based responses, include thigmo differentiation, such as the induction of invasion structures by plant pathogens in response to topography; thigmonasty, where contact with a motile prey rapidly triggers its capture; and thigmotropism, where the direction of hyphal growth is guided by physical features in the environment. Like plants and some bacteria, fungi grow as walled cells. Despite the well-demonstrated importance of thigmo responses in numerous stages of fungal growth and development, it is not known how fungal cells sense contact through the relatively rigid structure of the cell wall. However, while sensing mechanisms at the molecular level are not entirely understood, the downstream signaling pathways that are activated by contact sensing are being elucidated. In the majority of cases, the response to contact is complemented by chemical cues and both are required, either sequentially or simultaneously, to elicit normal developmental responses. The importance of a sense of touch in the lifestyles and development of diverse fungi is highlighted in this review, and the candidate molecular mechanisms that may be involved in fungal contact sensing are discussed.

  15. Histopathology of fungal diseases of the lung.

    Science.gov (United States)

    Roden, Anja C; Schuetz, Audrey N

    2017-11-01

    Fungal pneumonias can be a diagnostic problem. However, their recognition is important as they can pose a significant health risk, especially in the immunocompromised host. While many of these infections are accompanied by necrotizing or non-necrotizing granulomas, some might be characterized by cellular interstitial pneumonia, intra-alveolar frothy material or only minimal inflammatory change. Much of the tissue reaction is dependent on the immune status of the patient and the type of fungal organism. While many of the fungi can be identified in tissue, especially if using histochemical stains such as Grocott's Methenamine Silver (GMS) stain and/or Periodic Acid Schiff (PAS) stain, in some cases, these stains are negative and the organisms can only be identified in cultures or using special techniques such as PCR or fungal serology. Some fungi can be accurately identified in tissue based on morphologic features; others require culture for exact classification. Knowledge about immune status, geographic region and social history of the patient are helpful in identifying the fungus and, therefore, detailed clinical and travel histories are important. In this manuscript we aim to describe the most common fungal infections that occur in the lung, their morphologic features, and differential diagnoses. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Grass fungal endophytes and uses thereof

    Energy Technology Data Exchange (ETDEWEB)

    Craven, Kelly

    2015-03-10

    The invention provides isolated fungal endophytes and synthetic combinations thereof with host grass plants. Methods for inoculating grass plant with the endophytes, for propagating the grass-endophyte combinations, and for producing feeds and biofuels from grass-endophyte combinations are also provided.

  17. Fungal Planet description sheets: 400-468

    Czech Academy of Sciences Publication Activity Database

    Crous, P.W.; Wingfield, M. J.; Richardson, D. M.; Le Roux, J. J.; Strasberg, D.; Edwards, J.; Roets, F.; Hubka, V.; Taylor, P.W.J.; Heykoop, M.; Martín, M.P.; Moreno, G.; Sutton, D.A.; Wiederhold, N.P.; Barnes, C.W.; Carlavilla, J.R.; Gené, J.; Giraldo, A.; Guarnaccia, V.; Guarro, J.; Hernández-Restrepo, M.; Kolařík, Miroslav; Manjón, J.L.; Pascoe, I.G.; Popov, E.S.; Sandoval-Denis, M.; Woudenberg, J.H.C.; Acharya, K.; Alexandrova, A.V.; Alvarado, P.; Barbosa, R.N.; Baseia, I.G.; Blanchette, R.A.; Boekhout, T.; Burgess, T.I.; Cano-Lira, J.F.; Čmoková, A.; Dimitrov, R.A.; Dyakov, M.Yu.; Dueñas, M.; Dutta, A.K.; Esteve- Raventós, F.; Fedosova, A.G.; Fournier, J.; Gamboa, P.; Gouliamova, D.E.; Grebenc, T.; Groenewald, M.; Hanse, B.; Hardy, G.E.St.J.; Held, B.W.; Jurjević, Ž.; Kaewgrajang, T.; Latha, K.P.D.; Lombard, L.; Luangsa-Ard, J.J.; Lysková, P.; Mallátová, N.; Manimohan, P.; Miller, A.N.; Mirabolfathy, M.; Morozova, O.V.; Obodai, M.; Oliveira, N.T.; Otto, E.C.; Paloi, S.; Peterson, S.W.; Phosri, C.; Roux, J.; Salazar, W.A.; Sánchez, A.; Sarria, G.A.; Shin, H.-D.; Silva, B.D.B.; Silva, G.A.; Smith, M.Th.; Souza-Motta, C.M.; Stchigel, A.M.; Stoilova-Disheva, M.M.; Sulzbacher, M.A.; Telleria, M.T.; Toapanta, C.; Traba, J.M.; Valenzuela-Lopez, N.; Watling, R.; Groenewald, J.Z.

    2016-01-01

    Roč. 36, July (2016), s. 316-458 ISSN 0031-5850 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61388971 Keywords : ITS DNA barcodes * LSU * fungal species Subject RIV: EE - Microbiology, Virology Impact factor: 7.511, year: 2016

  18. Plant Fungal Pathogens: Methods and Protocols

    NARCIS (Netherlands)

    Bolton, M.D.; Thomma, B.P.H.J.

    2012-01-01

    Over the course of evolution, fungi have adapted to occupy specific niches, from symbiotically inhabiting the flora of the intestinal tract of mammals to saprophytic growth on leaf litter resting on the forest floor. In Plant Fungal Pathogens: Methods and Protocols, expert researchers in the field

  19. Standard methods for fungal brood disease research

    DEFF Research Database (Denmark)

    Jensen, Annette Bruun; Aronstein, Kathrine; Manuel Flores, Jose

    2013-01-01

    Chalkbrood and stonebrood are two fungal diseases associated with honey bee brood. Chalkbrood, caused by Ascosphaera apis, is a common and widespread disease that can result in severe reduction of emerging worker bees and thus overall colony productivity. Stonebrood is caused by Aspergillus spp. ...

  20. Pre- and postharvest fungal apple diseases

    Science.gov (United States)

    The domesticated apple (Malus domestica) is the most significant pome fruit grown and consumed worldwide. China is the largest producer followed by the United States on a global scale. However, fungal plant pathogens cause significant economic losses in the field and in storage which negatively impa...